Preparation and Characterization of Fluorescent SiO2 Microspheres
NASA Astrophysics Data System (ADS)
Xu, Cui; Zhang, Hao; Guan, Ruifang
2018-01-01
Fluorescent compound without typical fluorophores was synthesized with citric acid (CA) and aminopropyltriethoxysilane (APTS) firstly, and then it was grafted to the surface of the prepared SiO2 microspheres by chemical reaction. The fluorescent SiO2 microspheres with good fluorescent properties were obtained by optimizing the reaction conditions. And the morphology and structure of the fluorescent SiO2 microspheres have been characterized by scanning electron microscopy (SEM) and fourier transform infrared (FTIR) spectroscopy. The results showed that the preparation of fluorescent SiO2 microspheres have good monodispersity and narrow particle size distribution. Moreover, the fluorescent SiO2 microspheres can be applied to detect Fe3+ in aqueous solution, prepare fluorescent SiO2 rubber, and have potential to be applied in the fluorescent labeling and fingerprint appearing technique fields.
NASA Technical Reports Server (NTRS)
Hubler, M.; Souders, J. E.; Shade, E. D.; Hlastala, M. P.; Polissar, N. L.; Glenny, R. W.
1999-01-01
The aim of the study was to validate a nonradioactive method for relative blood flow measurements in severely injured lungs that avoids labor-intensive tissue processing. The use of fluorescent-labeled microspheres was compared with the standard radiolabeled-microsphere method. In seven sheep, lung injury was established by using oleic acid. Five pairs of radio- and fluorescent-labeled microspheres were injected before and after established lung injury. Across all animals, 175 pieces were selected randomly. The radioactivity of each piece was determined by using a scintillation counter. The fluorescent dye was extracted from each piece with a solvent without digestion or filtering. The fluorescence was determined with an automated fluorescent spectrophotometer. Perfusion was calculated for each piece from both the radioactivity and fluorescence and volume normalized. Correlations between flow determined by the two methods were in the range from 0.987 +/- 0.007 (SD) to 0.991 +/- 0.002 (SD) after 9 days of soaking. Thus the fluorescent microsphere technique is a valuable tool for investigating regional perfusion in severely injured lungs and can replace radioactivity.
Martens, Andreas; Rojas, Sebastian V; Baraki, Hassina; Rathert, Christian; Schecker, Natalie; Hernandez, Sara Rojas; Schwanke, Kristin; Zweigerdt, Robert; Martin, Ulrich; Saito, Shunsuke; Haverich, Axel; Kutschka, Ingo
2014-01-01
The limited effectiveness of cardiac cell therapy has generated concern regarding its clinical relevance. Experimental studies show that cell retention and engraftment are low after injection into ischemic myocardium, which may restrict therapy effectiveness significantly. Surgical aspects and mechanical loss are suspected to be the main culprits behind this phenomenon. As current techniques of monitoring intramyocardial injections are complex and time-consuming, the aim of the study was to develop a fast and simple model to study cardiac retention and distribution following intramyocardial injections. For this purpose, our main hypothesis was that macroscopic fluorescence imaging could adequately serve as a detection method for intramyocardial injections. A total of 20 mice underwent ligation of the left anterior descending artery (LAD) for myocardial infarction. Fluorescent microspheres with cellular dimensions were used as cell surrogates. Particles (5 × 10(5)) were injected into the infarcted area of explanted resting hearts (Ex vivo myocardial injetions EVMI, n = 10) and in vivo into beating hearts (In vivo myocardial injections IVMI, n = 10). Microsphere quantification was performed by fluorescence imaging of explanted organs. Measurements were repeated after a reduction to homogenate dilutions. Cardiac microsphere retention was 2.78 × 10(5) ± 0.31 × 10(5) in the EVMI group. In the IVMI group, cardiac retention of microspheres was significantly lower (0.74 × 10(5) ± 0.18 × 10(5); p<0.05). Direct fluorescence imaging revealed venous drainage through the coronary sinus, resulting in a microsphere accumulation in the left (0.90 × 10(5) ± 0.20 × 10(5)) and the right (1.07 × 10(5) ± 0.17 × 10(5)) lung. Processing to homogenates involved further particle loss (p<0.05) in both groups. We developed a fast and simple direct fluorescence imaging method for biodistribution analysis which enabled the quantification of fluorescent microspheres after intramyocardial delivery using macroscopic fluorescence imaging. This new technique showed massive early particle loss and venous drainage into the right atrium leading to substantial accumulation of graft particles in both lungs.
Ge, Minghao; Bai, Pengli; Chen, Mingli; Tian, Jingjing; Hu, Jun; Zhi, Xu; Yin, Huancai; Yin, Jian
2018-03-01
Here, we utilized the ultrasonic emulsification technique to generate hyaluronic acid microspheres incorporating a fluorescence-based glucose biosensor. We synthesized a novel lanthanide ion luminophore based on Eu 3+ . Eu sulfosuccinimidyl dextran (Eu-dextran) and Alexa Fluor 647 sulfosuccinimidyl-ConA (Alexa Fluor 647-ConA) were encapsulated in hyaluronic acid hydrogel to generate microspheres. Glucose sensing was carried out using a fluorescence resonance energy transfer (FRET)-based assay principle. A proportional fluorescence intensity increase was found within a 0.5-10-mM glucose concentration range. The glucose-sensing strategy showed an excellent tolerance for potential interferents. Meanwhile, the fluorescent signal of hyaluronic acid microspheres was very stable after testing for 72 h in glucose solution. Overall, hyaluronic acid microspheres encapsulating sensing biomolecules offer a stable and biocompatible biosensor for a variety of applications including cell culture systems, tissue engineering, detection of blood glucose, etc. Graphical abstract We report an ingenious biosensor encapsulated in hyaluronic acid microspheres for monitoring of glucose. Glucose sensing is carried out using a fluorescence resonance energy transfer-based assay principle with a novel lanthanide ions luminophore. The glucose detection system has excellent biocompatibility and stability for monitoring of glucose.
Use of an automated fluorescent microsphere method to measure regional blood flow in the fetal lamb.
Tan, W; Riggs, K W; Thies, R L; Rurak, D W
1997-08-01
We have developed a method for measuring regional blood flow by means of fluorescent microspheres in all organs and tissues of the fetal lamb, including brain, heart, lung, liver, gut, spleen, kidney, adrenal, brown fat, skin, muscle, bone, and placenta. Five different fluorescent-labeled microspheres were used: blue (B), yellow-green (Y), orange (O), red (R), and crimson (C). An automated, 96-well microplate fluorescent reader (bottom reading) was chosen for the assay because of the rapidity and high throughput that it offers. Tissue samples were digested by 4 M ethanolic KOH. The sedimentation method and dye extraction with Cellosolve acetate, as previously reported by others, were used for the sample processing. The bones were crushed and allowed to directly soak in Cellosolve acetate to extract the dye. The relationship between microsphere number and fluorescent intensity was linear over a broad range of microsphere numbers (80-20,000/mL). The coefficients of variation of within-run and between-run precision were 3.39 +/- 1.10% and 4.54 +/- 1.10%, respectively. Recovery of microspheres from tissues and blood averaged 94.3 +/- 2.5% and was not dependent on microsphere number. The spillover of the fluorescent signals into adjacent colors was 4.0 +/- 0.1% for O to Y, 8.1 +/- 0.4% for O to R, and 9.1 +/- 0.5% for R to C, and these values were constant over a wide range in concentrations of the microsphere pairs. No evidence was obtained for quenching of the emission of one fluorophore via photon absorption by another fluorophore. The measurements of regional blood flow obtained with fluorescent microspheres in three chronically instrumented fetal lambs at approximately 140 days gestation were similar to the flow estimates obtained using radioactive microspheres in four other fetal lambs at the same gestational age. The fluorescent method is thus a viable alternative to the radioactive technique for the measurement of regional blood flow to all fetal organs and tissues, particularly when an automated fluorescent microplate reader is employed to reduce analysis time.
Poe, Bobby G; Navratil, Marian; Arriaga, Edgar A
2006-12-29
Flow cytometry (FCM) and more recently capillary electrophoresis with post-column laser-induced fluorescence detection (CE-LIF) have both been used for subcellular particle analysis but their analytical performance has not been compared. In this work, we compare a commercial FCM with an in-house built CE-LIF instrument using fluorescently labeled microspheres and isolated mitochondria. As evidenced by the relative standard deviation (RSD) of the individual fluorescence intensities, FCM is two-fold better than CE-LIF for microspheres with > or =1.5 x 10(6) molecules of equivalent soluble fluorescein (MESF). However, FCM has a comparatively low signal-to-noise ratio (S/N) and high RSD for microspheres with <1.5 x 10(6) MESF. CE-LIF, on the other hand, produces S/N ratios that are >25 times higher than FCM for all the microspheres tested and a lower RSD for microspheres with <1.5 x 10(6) MESF. When 10-N-nonyl acridine orange (NAO)-labeled mitochondria are analyzed, the S/N ratios of both techniques are similar. This appears to result from photobleaching of NAO-labeled mitochondria as they are detected by the LIF detector of the CE-LIF instrument. Both techniques have a niche in subcellular analysis; FCM has the advantage of collecting data for thousands of particles quickly, whereas CE-LIF consumes less than a nanoliter of sample and provides the electrophoretic mobility for individual particles.
A Simple and Inexpensive Technique for Assessing Microbial Contamination during Drilling Operations
NASA Astrophysics Data System (ADS)
Friese, A.; Kallmeyer, J.; Wagner, D.; Kitte, J. A.
2016-12-01
Exploration of the Deep Biosphere relies on drilling, which inevitably causes infiltration of drilling fluids, containing non-indigenous microbes from the surface, into the core. Therefore it is absolutely necessary to trace contamination of the sediment core in order to identify uncontaminated samples for microbiological investigations in drill core samples. To do this, usually a tracer is mixed into the drilling fluid. In past drilling operations a variety of tracers have been used including dyes, salts, dissolved gasses, and microspheres. The latter are microbe-sized fluorescent particles that can be detected with very high sensitivity. Each tracer has its specific strengths and weaknesses, for microspheres the main problem was the high price, which limited the use to spot checks or drilling operations that require only small amounts of drilling fluid. Here, we present a modified microsphere tracer approach, using an aqueous fluorescent pigment dispersion that has a similar concentration of fluorescent particles as previously used microsphere tracers. However, compared to previous microsphere tracers, the cost of the new tracer is four orders of magnitude lower, allowing for a much more liberal use even in large-scale operations. Its suitability for large drilling campaigns was successfully tested at the ICDP Deep Drilling at Lake Towuti, Sulawesi, Indonesia and at the ICDP Deep Drilling at Lake Chalco, Mexico. Contamination can be detected by fluorescence microscopy or by flow cytometry at a sensitivity that is in the range of established techniques. Quantification of the tracer thus only requires a minimum of equipment and by using a small portable cytometer, high-resolution data can be obtained directly on-site within minutes and with minimal effort. Therefore this approach offers an inexpensive but powerful alternative technique for contamination assessment for future drilling campaigns.
Luo, Guilin; Wang, Jianxin; Wang, Yingying; Feng, Bo; Weng, Jie
2015-01-01
Growth factor-loaded fluorescent alginate microspheres, which can realise sustained growth factor release and fluorescence imaging, were synthesised by in situ formation of ZnO quantum dots (QDs) and covalent graft of 4-(1-pyrenyl) butyric acid (PBA). BSA was chosen as a growth factor model protein to study the release kinetic of growth factors from alginate microspheres. The microsphere size and fluorescent properties were also investigated. Investigations of cell culture were used for evaluating biocompatibility of BSA-loaded fluorescent microspheres and fluorescence imaging property of ZnO QDs and PBA-grafted sodium alginate from the microspheres. The results show that they have good fluorescent property either to microspheres or to cells and fluorescent microspheres have good biocompatibility and property in sustained release of growth factors. The obtained microspheres will be expected to realise the imaging of cells and materials and also the release of growth factor in tissue engineering or in cell culture.
Han, Daehoon; Hong, Jinkee; Kim, Hyun Cheol; Sung, Jong Hwan; Lee, Jong Bum
2013-11-01
Many highly sensitive protein detection techniques have been developed and have played an important role in the analysis of proteins. Herein, we report a novel technique that can detect proteins sensitively and effectively using aptamer-based DNA nanostructures. Thrombin was used as a target protein and aptamer was used to capture fluorescent dye-labeled DNA nanobarcodes or thrombin on a microsphere. The captured DNA nanobarcodes were replaced by a thrombin and aptamer interaction. The detection ability of this approach was confirmed by flow cytometry with different concentrations of thrombin. Our detection method has great potential for rapid and simple protein detection with a variety of aptamers.
Protein specific fluorescent microspheres for labelling a protein
NASA Technical Reports Server (NTRS)
Rembaum, Alan (Inventor)
1982-01-01
Highly fluorescent, stable and biocompatible microspheres are obtained by copolymerizing an acrylic monomer containing a covalent bonding group such as hydroxyl, amine or carboxyl, for example, hydroxyethylmethacrylate, with an addition polymerizable fluorescent comonomer such as dansyl allyl amine. A lectin or antibody is bound to the covalent site to provide cell specificity. When the microspheres are added to a cell suspension the marked microspheres will specifically label a cell membrane by binding to a specific receptor site thereon. The labeled membrane can then be detected by fluorescence of the fluorescent monomer.
Song, Jing; Qiu, Tian; Chen, Yun; Zhang, Wei; Fan, Li-Juan
2015-08-15
The layer-by-layer (LBL) technique was employed for preparing fluorescent microspheres with a core-shell structure by the alternating adsorption of positively charged poly(p-phenylenevinylene) precursor (pre-PPV) and the polyanions onto polymer substrate spheres, followed by the thermal elimination to convert pre-PPV into fluorescent poly(p-phenylenevinylene) (PPV). Weak polyelectrolytes poly(acrylic acid) (PAA) (usually in a partly ionized form) and strong polyelectrolytes poly(sodium-p-styrenesulfonate) (PSS) were used as the anions to space the PPV layers and reduce the fluorescence self-quenching. Flow cytometry, combined with spectroscopy and microscopy, were used to study the structure and photophysical properties of the resulting microspheres. Optimization of the processing factors was carried out. PAA and PSS as weak and strong polyelectrolytes, respectively, displayed very different influence on the final emission of the spheres. Such difference was attributed to different inherent characteristics of PAA and PSS after detailed investigation in many aspects. In addition, the fluorescent spheres were found to have excellent photostability and thermal stability. Copyright © 2015 Elsevier Inc. All rights reserved.
Fabrication of Polymer Microspheres for Optical Resonator and Laser Applications.
Yamamoto, Yohei; Okada, Daichi; Kushida, Soh; Ngara, Zakarias Seba; Oki, Osamu
2017-06-02
This paper describes three methods of preparing fluorescent microspheres comprising π-conjugated or non-conjugated polymers: vapor diffusion, interface precipitation, and mini-emulsion. In all methods, well-defined, micrometer-sized spheres are obtained from a self-assembling process in solution. The vapor diffusion method can result in spheres with the highest sphericity and surface smoothness, yet the types of the polymers able to form these spheres are limited. On the other hand, in the mini-emulsion method, microspheres can be made from various types of polymers, even from highly crystalline polymers with coplanar, π-conjugated backbones. The photoluminescent (PL) properties from single isolated microspheres are unusual: the PL is confined inside the spheres, propagates at the circumference of the spheres via the total internal reflection at the polymer/air interface, and self-interferes to show sharp and periodic resonant PL lines. These resonating modes are so-called "whispering gallery modes" (WGMs). This work demonstrates how to measure WGM PL from single isolated spheres using the micro-photoluminescence (µ-PL) technique. In this technique, a focused laser beam irradiates a single microsphere, and the luminescence is detected by a spectrometer. A micromanipulation technique is then used to connect the microspheres one by one and to demonstrate the intersphere PL propagation and color conversion from coupled microspheres upon excitation at the perimeter of one sphere and detection of PL from the other microsphere. These techniques, µ-PL and micromanipulation, are useful for experiments on micro-optic application using polymer materials.
Atom-Photon Coupling from Nitrogen-vacancy Centres Embedded in Tellurite Microspheres
NASA Astrophysics Data System (ADS)
Ruan, Yinlan; Gibson, Brant C.; Lau, Desmond W. M.; Greentree, Andrew D.; Ji, Hong; Ebendorff-Heidepriem, Heike; Johnson, Brett C.; Ohshima, Takeshi; Monro, Tanya M.
2015-06-01
We have developed a technique for creating high quality tellurite microspheres with embedded nanodiamonds (NDs) containing nitrogen-vacancy (NV) centres. This hybrid method allows fluorescence of the NVs in the NDs to be directly, rather than evanescently, coupled to the whispering gallery modes of the tellurite microspheres at room temperature. As a demonstration of its sensing potential, shifting of the resonance peaks is also demonstrated by coating a sphere surface with a liquid layer. This new approach is a robust way of creating cavities for use in quantum and sensing applications.
Immunomicrospheres - Reagents for cell labeling and separation
NASA Technical Reports Server (NTRS)
Rembaum, A.; Dreyer, W. J.
1980-01-01
Immunomicrospheres are specially designed microscopic particles that have antibodies or similar molecules chemically bound to their surfaces. The antibody-coated microspheres react in a highly specific way with target cells, viruses, or other antigenic agents. Immunomicrospheres may be synthesized so that they incorporate compounds that are highly radioactive, intensely fluorescent, magnetic, electron opaque, highly colored, or pharmacologically active. These various types of microspheres may be coated with pure, highly specific monoclonal antibodies obtained by the new hybridoma cell cloning techniques or with conventional antibody preparations. Some of the many present and potential applications for these new reagents are (1) new types of radioimmune or immunofluorescent assays, (2) improved fluorescence microscopy, (3) separation of cells on the basis of the fluorescent, electrophoretic, or magnetic properties of bound immunomicrospheres, (4) markers for use in several types of electron or standard light microscopy, and (5) delivery of lethal compouds to specific undesirable living cells. The combination of the various new types of synthetic microspheres and the newly available homogeneous antibodies offers new opportunities in research, diagnosis, and therapy.
NASA Astrophysics Data System (ADS)
Ryu, Inkeon; Kim, Daekeun
2018-04-01
A typical selective plane illumination microscopy (SPIM) image size is basically limited by the field of view, which is a characteristic of the objective lens. If an image larger than the imaging area of the sample is to be obtained, image stitching, which combines step-scanned images into a single panoramic image, is required. However, accurately registering the step-scanned images is very difficult because the SPIM system uses a customized sample mount where uncertainties for the translational and the rotational motions exist. In this paper, an image registration technique based on multiple fluorescent microsphere tracking is proposed in the view of quantifying the constellations and measuring the distances between at least two fluorescent microspheres embedded in the sample. Image stitching results are demonstrated for optically cleared large tissue with various staining methods. Compensation for the effect of the sample rotation that occurs during the translational motion in the sample mount is also discussed.
Nie, Shuai; Benito-Peña, Elena; Zhang, Huaibin; Wu, Yue; Walt, David R
2013-10-10
Herein, we describe a protocol for simultaneously measuring six proteins in saliva using a fiber-optic microsphere-based antibody array. The immuno-array technology employed combines the advantages of microsphere-based suspension array fabrication with the use of fluorescence microscopy. As described in the video protocol, commercially available 4.5 μm polymer microspheres were encoded into seven different types, differentiated by the concentration of two fluorescent dyes physically trapped inside the microspheres. The encoded microspheres containing surface carboxyl groups were modified with monoclonal capture antibodies through EDC/NHS coupling chemistry. To assemble the protein microarray, the different types of encoded and functionalized microspheres were mixed and randomly deposited in 4.5 μm microwells, which were chemically etched at the proximal end of a fiber-optic bundle. The fiber-optic bundle was used as both a carrier and for imaging the microspheres. Once assembled, the microarray was used to capture proteins in the saliva supernatant collected from the clinic. The detection was based on a sandwich immunoassay using a mixture of biotinylated detection antibodies for different analytes with a streptavidin-conjugated fluorescent probe, R-phycoerythrin. The microarray was imaged by fluorescence microscopy in three different channels, two for microsphere registration and one for the assay signal. The fluorescence micrographs were then decoded and analyzed using a homemade algorithm in MATLAB.
NASA Astrophysics Data System (ADS)
Charron, Luc; Harmer, Andrea; Lilge, Lothar
2005-09-01
A technique to produce fluorescent cell phantom standards based on calcium alginate microspheres with encapsulated fluorescein-labeled dextrans is presented. An electrostatic ionotropic gelation method is used to create the microspheres which are then exposed to an encapsulation method using poly-l-lysine to trap the dextrans inside. Both procedures were examined in detail to find the optimal parameters producing cell phantoms meeting our requirements. Size distributions favoring 10-20 microns microspheres were obtained by varying the high voltage and needle size parameters. Typical size distributions of the samples were centered at 150 μm diameter. Neither the molecular weight nor the charge of the dextrans had a significant effect on their retention in the microspheres, though anionic dextrans were chosen to help in future capillary electrophoresis work. Increasing the exposure time of the microspheres to the poly-l-lysine solution decreased the leakage rates of fluorescein-labeled dextrans.
Atom–Photon Coupling from Nitrogen-vacancy Centres Embedded in Tellurite Microspheres
Ruan, Yinlan; Gibson, Brant C.; Lau, Desmond W. M.; Greentree, Andrew D.; Ji, Hong; Ebendorff-Heidepriem, Heike; Johnson, Brett C.; Ohshima, Takeshi; Monro, Tanya M.
2015-01-01
We have developed a technique for creating high quality tellurite microspheres with embedded nanodiamonds (NDs) containing nitrogen-vacancy (NV) centres. This hybrid method allows fluorescence of the NVs in the NDs to be directly, rather than evanescently, coupled to the whispering gallery modes of the tellurite microspheres at room temperature. As a demonstration of its sensing potential, shifting of the resonance peaks is also demonstrated by coating a sphere surface with a liquid layer. This new approach is a robust way of creating cavities for use in quantum and sensing applications. PMID:26095793
Bronchial circulation in the marsupial opossum, Didelphis marsupialis.
Bernard, S L; Luchtel, D L; Glenny, R W; Lakshminarayan, S
1996-08-01
This study characterizes the existence of a bronchial circulation in a marsupial, an animal which does not undergo placental development and does not have a ductus arteriosus. Direct perfusion of the lung by the pulmonary vasculature during the fetal development of opossums may occur, potentially eliminating the need for a bronchial circulation. We used radio- and fluorescent-labeled microspheres in conjunction with postmortem intravascular casting to determine if opossums have a systemic (bronchial) blood supply to the lung (n = 9). Gross postmortem examination of the intravascular casts showed a well-developed common bronchial artery. The histological distribution pattern of fluorescent microspheres was primarily to the airways. A few fluorescent microspheres were observed in the alveolar capillaries, indicating that a precapillary bronchial-to-pulmonary anastomosis exists in the opossum. Using the reference flow technique, total bronchial blood flow to the left lung averaged 0.95 +/- 0.58 SE ml/min. The presence of a bronchial circulation in the opossum suggests that it is more than a vestigial structure from embryonic development, potentially supporting its functional importance for carrying nutrients to the airway.
Braune, Anja; Scharffenberg, Martin; Naumann, Anne; Bluth, Thomas; de Abreu, Marcelo Gama; Kotzerke, Jörg
2018-06-01
We compared 68 Gallium ( 68 Ga)- and fluorescence-labeled microspheres for measurement of pulmonary perfusion distribution in anesthetized pigs without lung injury. In two mechanically ventilated pigs, the distribution of pulmonary perfusion was marked in vivo with 68 Ga- and fluorescence-labeled microspheres in supine and prone position. After each injection, the distribution of 68 Ga-labeled microspheres was measured in vivo with positron emission tomography/ computed tomography (PET/CT) in the position in which microspheres were injected and vice versa. The distribution of fluorescence-labeled microspheres was measured ex vivo . Perfusion distributions were compared between methods and postures within four lung regions and along the ventro-dorsal gradient. After each injection of 68 Ga-labeled microspheres, changes in ventro-dorsal perfusion gradients induced by repositioning were compared for volume- and mass-normalized PET/CT measurements. Regional and gradient analyses of in vivo and ex vivo measurements, respectively, consistently revealed higher pulmonary perfusion in dorsal than ventral regions in supine positioned animals. Both methods showed more pronounced perfusion gradients in supine compared to prone position. Changes in animal position were associated with alterations in the ventro-dorsal perfusion gradient when volume-, but not mass-normalization was conducted for PET/CT data. Ex vivo fluorescence- and in vivo 68 Ga-labeled microspheres measurements revealed similar perfusion distributions. Mass-normalized perfusion measurements by 68 Ga-labeled microspheres and PET/CT were not affected by positioning artifacts. Schattauer GmbH.
Microsphere-aided optical microscopy and its applications for super-resolution imaging
NASA Astrophysics Data System (ADS)
Upputuri, Paul Kumar; Pramanik, Manojit
2017-12-01
The spatial resolution of a standard optical microscope (SOM) is limited by diffraction. In visible spectrum, SOM can provide ∼ 200 nm resolution. To break the diffraction limit several approaches were developed including scanning near field microscopy, metamaterial super-lenses, nanoscale solid immersion lenses, super-oscillatory lenses, confocal fluorescence microscopy, techniques that exploit non-linear response of fluorophores like stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, etc. Recently, photonic nanojet generated by a dielectric microsphere was used to break the diffraction limit. The microsphere-approach is simple, cost-effective and can be implemented under a standard microscope, hence it has gained enormous attention for super-resolution imaging. In this article, we briefly review the microsphere approach and its applications for super-resolution imaging in various optical imaging modalities.
A simple and inexpensive technique for assessing microbial contamination during drilling operations
NASA Astrophysics Data System (ADS)
Friese, André; Vuillemin, Aurèle; Kallmeyer, Jens; Wagner, Dirk
2016-04-01
Exploration of the Deep Biosphere relies on drilling, which inevitably causes infiltration of drilling fluids, containing allochthonous microbes from the surface, into the core. Therefore it is absolutely necessary to trace contamination of the sediment core in order to identify uncontaminated samples for microbiological investigations. Several techniques have been used in the past, including fluorescent dyes, perfluorocarbon tracers and fluorescent microspheres. Fluorescent dyes are inexpensive and easy to analyze on-site but are sensitive to light, pH and water chemistry. Furthermore, significant sorption to clays can decrease the fluorescence signal. Perfluorocarbon tracers are chemically inert hydrophobic compounds that can be detected with high sensitivity via gas chromatography, which might be a problem for on-site analysis. Samples have to be taken immediately after core retrieval as otherwise the volatile tracer will have diffused out of the core. Microsphere tracers are small (0.2 - 0.5 μm diameter) fluorescent plastic particles that are mixed into the drilling fluid. For analysis, these particles can be extracted from the sediment sample, transferred onto a filter and quantified via fluorescence microscopy. However, they are very expensive and therefore unsuitable for deep drilling operations that need large amounts of drilling fluids. Here, we present an inexpensive contamination control approach using fluorescent pigments initially used for coloring plastics. The price of this tracer is nearly three orders of magnitude lower than conventional microsphere tracers. Its suitability for large drilling campaigns was tested at the ICDP Deep Drilling at Lake Towuti, Sulawesi, Indonesia. The tracer was diluted 1:1000 in lake water, which was used as the drilling fluid. Additionally, a plastic bag filled with 20 mL of undiluted tracer was attached to the core catcher to increase the amount of particles in the liner fluid right at the core. After core retrieval, the core was cut and the liner fluid collected. From each whole round core (WRC) that was taken for microbiological and biogeochemical analyses, small samples of 1 cc were retrieved with sterile cutoff syringes from the rim, the center and an intermediate position. After dilution and homogenization in 9 mL MilliQ water, 10 μL of the sediment slurry was transferred onto a filter membrane and particles counted via fluorescence microscopy. Additionally, particles in the liner fluid were also quantified. This allows the quantification of the amount of drilling fluid that has entered the sediment sample during drilling. The minimum detectable volume of drilling fluid was in the order of single nanoliters per cc of sediment, which is in the range of established techniques. The presented method requires only a minimum of equipment and allows rapid determination of contamination in the sediment core and an easy to handle on-site analysis at low costs. The sensitivity is in the same range as perfluorocarbon and microsphere tracer applications. Thus, it offers an inexpensive but powerful technique for contamination assessment for future drilling campaigns.
Artz, Nathan S; Wentland, Andrew L; Sadowski, Elizabeth A; Djamali, Arjang; Grist, Thomas M; Seo, Songwon; Fain, Sean B
2011-02-01
The purpose of this study was to assess the ability of a flow-sensitive alternating inversion recovery-arterial spin labeling (FAIR-ASL) technique to track renal perfusion changes during pharmacologic and physiologic alterations in renal blood flow using microspheres as a gold standard. Fluorescent microsphere and FAIR-ASL perfusion were compared in the cortex of the kidney for 11 swine across 4 interventional time points: (1) under baseline conditions, (2) during an acetylcholine and fluid bolus challenge to increase perfusion, (3) initially after switching to isoflurane anesthesia, and (4) after 2 hours of isoflurane anesthesia. In 10 of the 11 swine, a bag of ice was placed on the hilum of 1 kidney at the beginning of isoflurane administration to further reduce perfusion in 1 kidney. Both ASL and microspheres tracked the expected cortical perfusion changes (P < 0.02) across the interventions, including an increase in perfusion during the acetylcholine challenge and decrease during the administration of isoflurane. Both techniques also measured lower cortical perfusion in the iced compared with the non-iced kidneys (P ≤ 0.01). The ASL values were systematically lower compared with microsphere perfusion. Very good correlation (r = 0.81, P < 0.0001) was observed between the techniques, and the relationship appeared linear for perfusion values in the expected physiologic range (microsphere perfusion <550 mL/min/100 g) although ASL values saturated for perfusion >550 mL/min/100 g. Cortical perfusion measured with ASL correlated with microspheres and reliably detected changes in renal perfusion in response to physiologic challenge.
NASA Astrophysics Data System (ADS)
Wang, Peng; Ebeling, Carl G.; Gerton, Jordan; Menon, Rajesh
In this paper, we demonstrate hyper-spectral imaging of fluorescent microspheres in a scanning-confocal-fluorescence microscope by spatially dispersing the spectra using a novel broadband diffractive optic, and applying a nonlinear optimization technique to extract the full-incident spectra. This broadband diffractive optic has a designed optical efficiency of over 90% across the entire visible spectrum. We used this technique to create two-color images of two fluorophores and also extracted their emission spectra with good fidelity. This method can be extended to image both spatially and spectrally overlapping fluorescent samples. Full control in the number of emission spectra and the feasibility of enhanced imaging speed are demonstrated as well.
Tsukada, Naoki; Katsumata, Masahiro; Oki, Koichi; Minami, Kazushi; Abe, Takato; Takahashi, Shinichi; Itoh, Yoshiaki; Suzuki, Norihiro
2018-01-15
A hemodynamic mechanism has long been assumed to play an important role in watershed infarction. In recent years, however, clinical evidence has indicated that an embolic mechanism is involved. The mechanism by which emboli are trapped preferentially in watershed areas remains unclear. In the present study, we developed a mouse embolus model using fluorescent microspheres with different diameters and evaluated the role of the microspheres' diameters in the generation of a watershed-patterned distribution. We injected fluorescent microspheres of four different diameters (i.e., 13, 24, 40, and 69 μm) into the internal carotid artery of C57BL/6 mice either (1) without ligation of the common carotid artery (normal perfusion pressure model: NPPM) or (2) with ligation of the common carotid artery (low perfusion pressure model: LPPM). Left common carotid artery ligation induced reductions in local cerebral blood flow in both the periphery and the core area of the left middle cerebral artery. A greater reduction in the border-zone area between the left anterior cerebral artery and the middle cerebral artery was also noted. After 24 h, the brains were removed and the distribution of the microspheres in the brain was evaluated using a fluorescence microscope. The 24-μm microspheres were distributed in the watershed area more frequently than the other microsphere sizes (P < .05, ANOVA followed by Tukey's test). Meanwhile, the distribution rates were similar between the NPPM and LPPM models for all microsphere sizes. This study suggested that the distribution pattern of the microspheres was only affected by the microspheres' diameters. Copyright © 2017 Elsevier B.V. All rights reserved.
Characterization of Fluorescent Polystyrene Microspheres for Advanced Flow Diagnostics
NASA Technical Reports Server (NTRS)
Maisto, Pietro M. F.; Lowe, K. Todd; Byun, Guibo; Simpson, Roger; Vercamp, Max; Danley, Jason E.; Koh, Brian; Tiemsin, Pacita; Danehy, Paul M.; Wohl, Christopher J.
2013-01-01
Fluorescent dye-doped polystyrene latex microspheres (PSLs) are being developed for velocimetry and scalar measurements in variable property flows. Two organic dyes, Rhodamine B (RhB) and dichlorofluorescence (DCF), are examined to assess laser-induced fluorescence (LIF) properties for flow imaging applications and single-shot temperature measurements. A major interest in the current research is the application of safe dyes, thus DCF is of particular interest, while RhB is used as a benchmark. Success is demonstrated for single-point laser Doppler velocimetry (LDV) and also imaging fluorescence, excited via a continuous wave 2 W laser beam, for exposures down to 10 ms. In contrast, when exciting with a pulsed Nd:YAG laser at 200 mJ/pulse, no fluorescence was detected, even when integrating tens of pulses. We show that this is due to saturation of the LIF signal at relatively low excitation intensities, 4-5 orders of magnitude lower than the pulsed laser intensity. A two-band LIF technique is applied in a heated jet, indicating that the technique effectively removes interfering inputs such as particle diameter variation. Temperature measurement uncertainties are estimated based upon the variance measured for the two-band LIF intensity ratio and the achievable dye temperature sensitivity, indicating that particles developed to date may provide about +/-12.5 C precision, while future improvements in dye temperature sensitivity and signal quality may enable single-shot temperature measurements with sub-degree precision.
NASA Technical Reports Server (NTRS)
1990-01-01
Vital information on a person's physical condition can be obtained by identifying and counting the population of T-cells and B-cells, lymphocytes of the same shape and size that help the immune system protect the body from the invasion of disease. The late Dr. Alan Rembaum developed a method for identifying the cells. The method involved tagging the T-cells and B-cells with microspheres of different fluorescent color. Microspheres, which have fluorescent dye embedded in them, are chemically treated so that they can link with antibodies. With the help of a complex antibody/antigen reaction, the microspheres bind themselves to specific 'targets,' in this case the T-cells or B-cells. Each group of cells can then be analyzed by a photoelectronic instrument at different wavelengths emitted by the fluorescent dyes. Same concept was applied to the separation of cancer cells from normal cells. Microspheres were also used to conduct many other research projects. Under a patent license Magsphere, Inc. is producing a wide spectrum of microspheres on a large scale and selling them worldwide for various applications.
NASA Astrophysics Data System (ADS)
Proctor, Ashley R.; Ramirez, Gabriel A.; Han, Songfeng; Liu, Ziping; Bubel, Tracy M.; Choe, Regine
2018-03-01
Nicotinamide has been shown to affect blood flow in both tumor and normal tissues, including skeletal muscle. Intraperitoneal injection of nicotinamide was used as a simple intervention to test the sensitivity of noninvasive diffuse correlation spectroscopy (DCS) to changes in blood flow in the murine left quadriceps femoris skeletal muscle. DCS was then compared with the gold-standard fluorescent microsphere (FM) technique for validation. The nicotinamide dose-response experiment showed that relative blood flow measured by DCS increased following treatment with 500- and 1000-mg / kg nicotinamide. The DCS and FM technique comparison showed that blood flow index measured by DCS was correlated with FM counts quantified by image analysis. The results of this study show that DCS is sensitive to nicotinamide-induced blood flow elevation in the murine left quadriceps femoris. Additionally, the results of the comparison were consistent with similar studies in higher-order animal models, suggesting that mouse models can be effectively employed to investigate the utility of DCS for various blood flow measurement applications.
Fang, Shiyue; Guan, Yousheng; Blatchley, Ernest R; Shen, Chengyue; Bergstrom, Donald E
2008-03-01
( E)-5-[2-(Methoxycarbonyl)ethenyl]cytidine was biotinylated through a diisopropylsilylacetal linkage and attached to the surface of hydrophilic streptavidin-coated microspheres through the high-affinity noncovalent interaction between biotin and streptavidin. The functionalized microspheres form a stable suspension in water. Upon UV irradiation, the nonfluorescent ( E)-5-[2-(methoxycarbonyl)ethenyl]cytidine on the microspheres undergoes photocyclization to produce highly fluorescent 3-beta-D-ribofuranosyl-2,7-dioxopyrido[2,3-d]pyrimidine. The fluorescence intensity of the microspheres can be correlated to the particle-specific UV doses applied at different suspension concentrations. The microspheres allow one to measure the UV dose (fluence) distribution in high-throughput water disinfection systems.
Synthesis of di-functional ligand and fluorescently labeling SiO2 microspheres
NASA Astrophysics Data System (ADS)
Chen, Kexu; Kang, Ming; Liu, Min; Shen, Simin; Sun, Rong
2018-05-01
In order to complete the fluorescent labeling of SiO2 microspheres, a kind of di-functional ligand was synthesized and purified, which could not only coordinate rare earth ions but also react with the active groups to bond host materials with an alkoxysilane groups. Fourier transform infrared spectroscopy (FT-IR), 1H NMR spectra, MS spectra, field emission scanning electron microscope (FESEM), transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS) and luminescence spectrophotometer were used to study the structure of di-functional ligand and properties of fluorescent coupling agent and fluorescent labeled SiO2 microspheres. The optimal experiment conditions were acquired as follows: molar ratio as 1: 4 (MDBM: MICPTES), reaction time at 6 h and reaction temperature as 65 °C (yield up to 40%) through the orthogonal experiment and purification process. The results indicated that fluorescent coupling agent presented red photoluminesence of Eu3+ ions at 610 nm, and the absolute quantum yield was 11%. On the other hand, the hydrolysis of the coupling agent reacted on the surface of SiO2 microspheres and presented fluorescent labeling homogeneously.
Tian, Ji-Yuan; Sun, Xiu-Qin; Chen, Xi-Guang
2008-05-01
Oral delivery of plasmid DNA (pDNA) is a desirable approach for fish immunization in intensive culture. However, its effectiveness is limited because of possible degradation of pDNA in the fish's digestive system. In this report, alginate microspheres loaded with pDNA coding for fish lymphocystis disease virus (LCDV) and green fluorescent protein were prepared with a modified oil containing water (W/O) emulsification method. Yield, loading percent and encapsulation efficiency of alginate microspheres were 90.5%, 1.8% and 92.7%, respectively. The alginate microspheres had diameters of less than 10 microm, and their shape was spherical. As compared to sodium alginate, a remarkable increase of DNA-phosphodiester and DNA-phosphomonoester bonds was observed for alginate microspheres loaded with pDNA by Fourier transform infrared (FTIR) spectroscopic analysis. Agarose gel electrophoresis showed a little supercoiled pDNA was transformed to open circular and linear pDNA during encapsulation. The cumulative release of pDNA in alginate microspheres was
NASA Astrophysics Data System (ADS)
Hennessy, Ricky; Koo, Chiwan; Ton, Phuc; Han, Arum; Righetti, Raffaella; Maitland, Kristen C.
2011-03-01
Ultrasound poroelastography can quantify structural and mechanical properties of tissues such as stiffness, compressibility, and fluid flow rate. This novel ultrasound technique is being explored to detect tissue changes associated with lymphatic disease. We have constructed a macroscopic fluorescence imaging system to validate ultrasonic fluid flow measurements and to provide high resolution imaging of microfluidic phantoms. The optical imaging system is composed of a white light source, excitation and emission filters, and a camera with a zoom lens. The field of view can be adjusted from 100 mm x 75 mm to 10 mm x 7.5 mm. The microfluidic device is made of polydimethylsiloxane (PDMS) and has 9 channels, each 40 μm deep with widths ranging from 30 μm to 200 μm. A syringe pump was used to propel water containing 15 μm diameter fluorescent microspheres through the microchannels, with flow rates ranging from 0.5 μl/min to 10 μl/min. Video was captured at a rate of 25 frames/sec. The velocity of the microspheres in the microchannels was calculated using an algorithm that tracked the movement of the fluorescent microspheres. The imaging system was able to measure particle velocities ranging from 0.2 mm/sec to 10 mm/sec. The range of flow velocities of interest in lymph vessels is between 1 mm/sec to 10 mm/sec; therefore our imaging system is sufficient to measure particle velocity in phantoms modeling lymphatic flow.
Wang, Jixiang; Wang, Yunyun; Qiu, Hao; Sun, Lin; Dai, Xiaohui; Pan, Jianming; Yan, Yongsheng
2017-01-01
Fluorescent molecularly imprinted polymers have shown great promise in biological or chemical separations and detection, due to their high stability, selectivity and sensitivity. In this work, fluorescent molecularly imprinted microsphere was synthesized via precipitation polymerization, which could separate efficiently and rapidly detect τ-fluvalinate (a toxic insecticide) in water samples, was reported. The fluorescent imprinted sensor showed excellent stability, outstanding selectivity and the limit of detection low to 12.14 nM, good regeneration ability which still kept good sensitivity after 8 cycling experiments and fluorescence quenching mechanism was illustrated in details. In addition, the fluorescent sensor was further used to detect τ-fluvalinate in real samples from Taihu Lake. Despite the relatively complex components of the environment water, the fluorescent imprinted microspheres sitll showed good recovery, clearly demonstrating the potental value of this smart sensor nanomaterial in environment monitoring. PMID:28485402
Histological methods to determine blood flow distribution with fluorescent microspheres.
Luchtel, D L; Boykin, J C; Bernard, S L; Glenny, R W
1998-11-01
We evaluated several histological methods and determined their advantages and disadvantages for histological studies of tissues and organs perfused with fluorescent microspheres. Microspheres retained their fluorescence in 7-10 microm serial sections with a change in the antimedium from toluene when samples were fixed in formalin and embedded in paraffin. Several antimedia allowed both wax infiltration of tissue and preservation of microsphere fluorescence. Histoclear II was the best substitute for toluene. When samples were fixed in formalin and embedded in glycol methacrylate, thinner (3-5 microm) sections provided greater histological detail but had fewer microspheres per section. Air dried lung tissue followed by Vibratome sectioning provided thick sections (100 microm) that facilitated rapid survey of large volumes of tissue for microspheres but limited histological detail, and the air drying procedure was restricted to lung tissue. Samples fixed in formalin followed by Vibratome sectioning of unembedded tissue provided better histological detail of lung tissue and was also useful for other organs. These sections were more difficult to handle and to mount on slides compared to air dried tissue, whereas fixed tissue embedded in gelatin provided better tissue support for Vibratome sectioning. Rapid freezing followed by cryo-microtome sectioning resulted in frozen sections that were relatively difficult to handle compared to embedded or unembedded tissue; they also deteriorated relatively rapidly with time. Paraffin sections were stained with hematoxylin and eosin or with aqueous methyl green, although tissue autofluorescence by itself was usually sufficient to identify histological features. Methacrylate sections quenched tissue autofluorescence, and Lee's stain or Richardson's stain were used for staining sections. Toluene based mountants such as Cytoseal quenched fluorescence, particularly the red fluorescent microspheres. Aqueous based mountants such as Aquamount, Crystal/Mount, Fluoromount-G were substituted, although such preparations were not as permanent as Cytoseal mounted coverglasses and tended to cause fading of stained sections.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kumagai, Tsutaru, E-mail: kumagai.t.af@m.titech.ac.jp; Kishi, Tetsuo; Yano, Tetsuji
2015-03-21
Bubble-containing Nd{sup 3+}-doped tellurite glass microspheres were fabricated by localized laser heating technique to investigate their optical properties for use as microresonators. Fluorescence and excitation spectra measurements were performed by pumping with a tunable CW-Ti:Sapphire laser. The excitation spectra manifested several sharp peaks due to the conventional whispering gallery mode (WGM) when the pumping laser was irradiated to the edge part of the microsphere. However, when the excitation light was irradiated on the bubble position inside the microsphere, “non-WGM excitation” was induced, giving rise to numerous peaks at a broad wavelength range in the excitation spectra. Thus, efficient excitation wasmore » achieved over a wide wavelength range. Lasing threshold excited at the bubble position was much lower than that for the excitation at the edges of the microsphere. The lowest value of the laser threshold was 34 μW for a 4 μm sphere containing a 0.5 μm bubble. Efficiency of the excitation at the bubble position with broadband light was calculated to be 5 times higher than that for the edge of the microsphere. The bubble-containing microsphere enables efficient utilization of broadband light excitation from light-emitting diodes and solar light.« less
Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.
2017-01-01
Though structured illumination (SI) microscopy is a popular imaging technique conventionally associated with fluorescent super-resolution, recent works have suggested its applicability towards sub-diffraction resolution coherent imaging with quantitative endogenous biological contrast. Here, we demonstrate that SI can efficiently integrate together the principles of fluorescent super-resolution and coherent synthetic aperture to achieve 3D dual-modality sub-diffraction resolution, fluorescence and refractive-index (RI) visualizations of biological samples. We experimentally demonstrate this framework by introducing a SI microscope capable of 3D sub-diffraction resolution fluorescence and RI imaging, and verify its biological visualization capabilities by experimentally reconstructing 3D RI/fluorescence visualizations of fluorescent calibration microspheres as well as alveolar basal epithelial adenocarcinoma (A549) and human colorectal adenocarcinmoa (HT-29) cells, fluorescently stained for F-actin. This demonstration may suggest SI as an especially promising imaging technique to enable future biological studies that explore synergistically operating biophysical/biochemical and molecular mechanisms at sub-diffraction resolutions. PMID:29296504
NASA Astrophysics Data System (ADS)
Kang, Yvonne Q.; François, Alexandre; Riesen, Nicolas; Monro, Tanya M.
2018-02-01
Whispering Gallery Mode (WGM) biosensors have been widely exploited over the past decade, owing to their unprecedented detection limits and label free capability. WGM based sensing mechanisms, such as resonance frequency shift, linewidth broadening, and splitting of the two counter-propagating WGMs, have been extensively researched and applied for bio-chemical sensing. However, the mode-splitting of the originally degenerate WGMs from different equatorial planes on a fluorescent microsphere has not been fully investigated. In this work, we break the symmetry of the surrounding environment outside the microsphere by partially embedding the sphere into a high-refractive-index medium (i.e. glue), to lift the degeneracy of the modes from different WGM planes. The split-modes from multiple planes of the fluorescent microsphere are indiscriminately collected. It is found that the effective quality factor Q of the WGMs increases non-conventionally as the Refractive Index (RI) of the probing liquid increases up to the point where it is equal to that of the glue. This presents a new methodology for quantifying changes in the probing environment based on the Q spoiling of the resonances as determined by the RI difference between the environment and that of the reference glue. Furthermore, we find that this sensing platform opens the door to simple self-referenced sensing techniques based on the analysis of the spectral positions of subsets of the split modes.
Savai, Rajkumar; Wolf, Joachim C.; Greschus, Susanne; Eul, Bastian G.; Schermuly, Ralph T.; Hänze, Jörg; Voswinckel, Robert; Langheinrich, Alexander C.; Grimminger, Friedrich; Traupe, Horst; Seeger, Werner; Rose, Frank
2005-01-01
In lung carcinomas the blood supply varies depending on tumor type and stage and can develop from pulmonary or bronchial circulation, or both. To examine this in vivo, primary bronchogenic Lewis lung carcinoma cells were intratracheally instilled in C57BL/6 mice. Within 7 days, histological examinations showed progressive tumor growth at the peripheral parenchymal region. The relative contribution of tumor blood supply via the pulmonary and systemic arteries was studied in detail using fluorescent microspheres (10 μm). When compared to healthy lung parenchyma (13:1), Lewis lung carcinoma tumor tissue (52:1) showed a fourfold increase in pulmonary to systemic microspheres, indicating that the pulmonary arteries are the predominant tumor-feeding vessels. After filling the vessels with a vascular cast, the microanatomy of vessels being derived from the pulmonary artery was visualized with micro computed tomography. Flat-panel volumetric computed tomography provided longitudinal visualization of tissue bridges between the growing tumor and the pulmonary vasculature. In this model of peripheral parenchymal malignancy, new imaging techniques allowed effective visualization of lung tumor growth and vascularization in living mice, demonstrating a pulmonary blood supply for lung tumors. PMID:16192630
Cryo-imaging of fluorescently labeled single cells in a mouse
NASA Astrophysics Data System (ADS)
Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.
2009-02-01
We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryoimaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 μm) and imaging, collecting color brightfield and fluorescent blockface image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [uT : heart (267 +/- 47.6 μm), liver (218 +/- 27.1 μm), brain (161 +/- 27.4 μm)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns. We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron-scale, fluorescence, and bright field image data. Here we describe our image preprocessing, analysis, and visualization techniques. Processing improves axial resolution, reduces subsurface fluorescence by 97%, and enables single cell detection and counting. High quality 3D volume renderings enable us to evaluate cell distribution patterns. Applications include the myriad of biomedical experiments using fluorescent reporter gene and exogenous fluorophore labeling of cells in applications such as stem cell regenerative medicine, cancer, tissue engineering, etc.
Preparation and flow cytometry of uniform silica-fluorescent dye microspheres.
Bele, Marjan; Siiman, Olavi; Matijević, Egon
2002-10-15
Uniform fluorescent silica-dye microspheres have been prepared by coating preformed monodispersed silica particles with silica layers containing rhodamine 6G or acridine orange. The resulting dispersions exhibit intense fluorescent emission between 500 and 600 nm, over a broad excitation wavelength range of 460 to 550 nm, even with exceedingly small amounts of dyes incorporated into the silica particles (10-30 ppm, expressed as weight of dye relative to weight of dry particles). The fluorescent particles can be prepared in micrometer diameters suitable for analyses using flow cytometry with 488-nm laser excitation.
Ahmed, J; Pulfer, M K; Linsenmeier, R A
2001-09-01
The most successful method for measuring absolute blood flow rate through the retinal circulation has been the use of radioactive microspheres. The purpose of this study was to develop a microsphere method that did not have the drawbacks associated with radioactivity and to use this method to make measurements of retinal blood flow in the cat. Blood flow measurements were made by injecting 15-microm-diameter polystyrene microspheres into the left ventricle of anesthetized, artificially ventilated cats. These microspheres were labeled with one of three fluorescent dyes. Retinal blood flow measurements were made by determining the number of spheres that were embedded in the retina and comparing them to the number found in a reference sample. Spheres in the retina were counted by making retinal whole mounts and taking retinal images with a CCD camera mounted on an epifluorescence microscope equipped with filter sets appropriate for imaging the dyes used to label the spheres. Blood flow measurements made under normal conditions showed a mean retinal blood flow of 19.8 +/- 12.4 ml/min 100 g tissue (mean +/- SD; n = 15 cats). Since the retinal circulation perfuses only the inner half of the retina, the effective flow rate in that region is about twice this value. RBF increased during hypoxemia (P(a)O2 < 42 mm Hg) to 336% of the normoxic value on average. Analysis of sphere deposition patterns showed that the central retina had a higher blood flow than the peripheral retina, although this difference was significant only during hypoxemia. We conclude that even with a relatively small number of spheres deposited in the retina, the technique can reveal important properties of the retinal circulation. Copyright 2001 Academic Press.
Juck, D F; Whissell, G; Steven, B; Pollard, W; McKay, C P; Greer, C W; Whyte, L G
2005-02-01
Fluorescent microspheres were applied in a novel fashion during subsurface drilling of permafrost and ground ice in the Canadian High Arctic to monitor the exogenous microbiological contamination of core samples obtained during the drilling process. Prior to each drill run, a concentrated fluorescent microsphere (0.5-microm diameter) solution was applied to the interior surfaces of the drill bit, core catcher, and core tube and allowed to dry. Macroscopic examination in the field demonstrated reliable transfer of the microspheres to core samples, while detailed microscopic examination revealed penetration levels of less than 1 cm from the core exterior. To monitor for microbial contamination during downstream processing of the permafrost and ground ice cores, a Pseudomonas strain expressing the green fluorescent protein (GFP) was painted on the core exterior prior to processing. Contamination of the processed core interiors with the GFP-expressing strain was not detected by culturing the samples or by PCR to detect the gfp marker gene. These methodologies were quick, were easy to apply, and should help to monitor the exogenous microbiological contamination of pristine permafrost and ground ice samples for downstream culture-dependent and culture-independent microbial analyses.
Juck, D. F.; Whissell, G.; Steven, B.; Pollard, W.; McKay, C. P.; Greer, C. W.; Whyte, L. G.
2005-01-01
Fluorescent microspheres were applied in a novel fashion during subsurface drilling of permafrost and ground ice in the Canadian High Arctic to monitor the exogenous microbiological contamination of core samples obtained during the drilling process. Prior to each drill run, a concentrated fluorescent microsphere (0.5-μm diameter) solution was applied to the interior surfaces of the drill bit, core catcher, and core tube and allowed to dry. Macroscopic examination in the field demonstrated reliable transfer of the microspheres to core samples, while detailed microscopic examination revealed penetration levels of less than 1 cm from the core exterior. To monitor for microbial contamination during downstream processing of the permafrost and ground ice cores, a Pseudomonas strain expressing the green fluorescent protein (GFP) was painted on the core exterior prior to processing. Contamination of the processed core interiors with the GFP-expressing strain was not detected by culturing the samples or by PCR to detect the gfp marker gene. These methodologies were quick, were easy to apply, and should help to monitor the exogenous microbiological contamination of pristine permafrost and ground ice samples for downstream culture-dependent and culture-independent microbial analyses. PMID:15691963
Preparation of chitosan/nano hydroxyapatite organic-inorganic hybrid microspheres for bone repair.
Chen, Jingdi; Pan, Panpan; Zhang, Yujue; Zhong, Shengnan; Zhang, Qiqing
2015-10-01
In this work, we encapsulated icariin (ICA) into chitosan (CS)/nano hydroxyapatite (nHAP) composite microspheres to form organic-inorganic hybrid microspheres for drug delivery carrier. The composition and morphology of composite microspheres were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and differential scanning calorimetry- thermogravimetric analysis (DSC-TGA). Moreover, we further studied the performance of swelling properties, degradation properties and drug release behavior of the microspheres. ICA, the extract of traditional Chinese medicine-epimedium, was combined to study drug release properties of the microspheres. ICA loaded microspheres take on a sustained release behavior, which can be not only ascribed to electrostatic interaction between reactive negative hydroxyl (OH) of ICA and positive amine groups (NH₂) of CS, but also depended on the homogeneous dispersion of HAP nanoparticles inside CS organic matrix. In addition, the adhesion and morphology of osteoblasts were detected by inverted fluorescence microscopy. The biocompatibility of CS/nHAP/ICA microspheres was evaluated by the MTT cytotoxicity assay, Hoechst 33258 and PI fluorescence staining. These studies demonstrate that composite microspheres provide a suitable microenvironment for osteoblast attachment and proliferation. It can be speculated that the ICA loaded CS-based organic-inorganic hybrid microspheres might have potential applications in drug delivery systems. Copyright © 2015 Elsevier B.V. All rights reserved.
Calibration of Wide-Field Deconvolution Microscopy for Quantitative Fluorescence Imaging
Lee, Ji-Sook; Wee, Tse-Luen (Erika); Brown, Claire M.
2014-01-01
Deconvolution enhances contrast in fluorescence microscopy images, especially in low-contrast, high-background wide-field microscope images, improving characterization of features within the sample. Deconvolution can also be combined with other imaging modalities, such as confocal microscopy, and most software programs seek to improve resolution as well as contrast. Quantitative image analyses require instrument calibration and with deconvolution, necessitate that this process itself preserves the relative quantitative relationships between fluorescence intensities. To ensure that the quantitative nature of the data remains unaltered, deconvolution algorithms need to be tested thoroughly. This study investigated whether the deconvolution algorithms in AutoQuant X3 preserve relative quantitative intensity data. InSpeck Green calibration microspheres were prepared for imaging, z-stacks were collected using a wide-field microscope, and the images were deconvolved using the iterative deconvolution algorithms with default settings. Afterwards, the mean intensities and volumes of microspheres in the original and the deconvolved images were measured. Deconvolved data sets showed higher average microsphere intensities and smaller volumes than the original wide-field data sets. In original and deconvolved data sets, intensity means showed linear relationships with the relative microsphere intensities given by the manufacturer. Importantly, upon normalization, the trend lines were found to have similar slopes. In original and deconvolved images, the volumes of the microspheres were quite uniform for all relative microsphere intensities. We were able to show that AutoQuant X3 deconvolution software data are quantitative. In general, the protocol presented can be used to calibrate any fluorescence microscope or image processing and analysis procedure. PMID:24688321
NASA Astrophysics Data System (ADS)
Nachman, Paul; Pinnick, R. G.; Hill, Steven C.; Chen, Gang; Chang, Richard K.; Mayo, Michael W.; Fernandez, Gilbert L.
1996-03-01
We report the design and operation of a prototype conditional-sampling spectrograph detection system that can record the fluorescence spectra of individual, micrometer-sized aerosols as they traverse an intense 488-nm intracavity laser beam. The instrument's image-intensified CCD detector is gated by elastic scattering or by undispersed fluorescence from particles that enter the spectrograph's field of view. It records spectra only from particles with preselected scattering-fluorescence levels (a fiber-optic-photomultiplier subsystem provides the gating signal). This conditional-sampling procedure reduces data-handling rates and increases the signal-to-noise ratio by restricting the system's exposures to brief periods when aerosols traverse the beam. We demonstrate these advantages by reliably capturing spectra from individual fluorescent microspheres dispersed in an airstream. The conditional-sampling procedure also permits some discrimination among different types of particles, so that spectra may be recorded from the few interesting particles present in a cloud of background aerosol. We demonstrate such discrimination by measuring spectra from selected fluorescent microspheres in a mixture of two types of microspheres, and from bacterial spores in a mixture of spores and nonfluorescent kaolin particles.
Integration of Microsphere Resonators with Bioassay Fluidics for Whispering Gallery Mode Imaging
Kim, Daniel C.; Armendariz, Kevin P.
2013-01-01
Whispering gallery mode resonators are small, radially symmetric dielectrics that trap light through continuous total internal reflection. The resonant condition at which light is efficiently confined within the structure is linked with refractive index, which has led to the development of sensitive label-free sensing schemes based on whispering gallery mode resonators. One resonator design uses inexpensive high index glass microspheres that offer intrinsically superior optical characteristics, but have proven difficult to multiplex and integrate with the fluidics for sample delivery and fluid exchange necessary for assay development. Recently, we introduced a fluorescence imaging approach that enables large scale multiplexing with microsphere resonators, thus removing one obstacle for assay development. Here we report an approach for microsphere immobilization that overcomes limitations arising from their integration with fluidic delivery. The approach is an adaptation of a calcium-assisted glass bonding method originally developed for microfluidic glass chip fabrication. Microspheres bonded to glass using this technique are shown to be stable with respect to fluid flow and show no detectable loss in optical performance. Measured Q-factors, for example, remain unchanged following sphere bonding to the substrate. The stability of the immobilized resonators is further demonstrated by transferring lipid films onto the immobilized spheres using the Langmuir-Blodgett technique. Bilayers of DOPC doped with GM1 were transferred onto immobilized resonators to detect the binding of cholera toxin to GM1. Binding curves generated from shifts in the whispering gallery mode resonance result in a measured Kd of 1.5 × 10−11 with a limit of detection of 3.3 pM. These results are discussed in terms of future assay development using microsphere resonators. PMID:23615457
USDA-ARS?s Scientific Manuscript database
A rapid lateral flow fluorescent microspheres immunochromatography test strip (FMs-ICTS) has been developed for the detection of aflatoxin M1 (AFM1) residues in milk. For this purpose, an ultra-sensitive anti-AFM1 monoclonal antibody (MAb) 1D3 was prepared and identified. The IC50 value of the MA...
Dao, Lam; Glancy, Brian; Lucotte, Bertrand; Chang, Lin-Ching; Balaban, Robert S; Hsu, Li-Yueh
2015-01-01
SUMMARY This paper investigates a post-processing approach to correct spatial distortion in two-photon fluorescence microscopy images for vascular network reconstruction. It is aimed at in vivo imaging of large field-of-view, deep-tissue studies of vascular structures. Based on simple geometric modeling of the object-of-interest, a distortion function is directly estimated from the image volume by deconvolution analysis. Such distortion function is then applied to sub volumes of the image stack to adaptively adjust for spatially varying distortion and reduce the image blurring through blind deconvolution. The proposed technique was first evaluated in phantom imaging of fluorescent microspheres that are comparable in size to the underlying capillary vascular structures. The effectiveness of restoring three-dimensional spherical geometry of the microspheres using the estimated distortion function was compared with empirically measured point-spread function. Next, the proposed approach was applied to in vivo vascular imaging of mouse skeletal muscle to reduce the image distortion of the capillary structures. We show that the proposed method effectively improve the image quality and reduce spatially varying distortion that occurs in large field-of-view deep-tissue vascular dataset. The proposed method will help in qualitative interpretation and quantitative analysis of vascular structures from fluorescence microscopy images. PMID:26224257
Cascaded Photoenhancement: Implications for Photonic Chemical and Biological Sensors
NASA Technical Reports Server (NTRS)
Fuller, Kirk A.; Smith, David D.
2006-01-01
Our analysis shows that coupling of gold nanoparticles to microspheres will evoke a cascading effect from the respective photoenhancement mechanisms. We refer to this amplification process as cascaded photoenhancement, and the resulting cavity amplification of surface-enhanced Raman scattering (SERS) and fluorescence as CASERS and CAF, respectively. Calculations, based on modal analysis of scattering and absorption by compound spheres, presented herein indicate that the absorption cross sections of metal nanoparticles immobilized onto dielectric microspheres can be greatly enhanced by cavity resonances in the microspheres without significant degradation of the resonators. Gain factors associated with CSP of 10(exp 3) - 10(exp 4) are predicted for realistic experimental conditions using homogenous microspheres. Cascaded surface photoenhancement thus has the potential of dramatically increasing the sensitivities of fluorescence and vibrational spectroscopies.
USDA-ARS?s Scientific Manuscript database
Background: The purpose of this study was the development of multiplex fluorescence microsphere immunoassay (FMIA) for the detection of Rift Valley fever virus (RVFV) IgG and IgM antibodies by incorporation of antigens from whole cell lysates and purified virions from MP12. Methods and Findings: Vir...
NASA Astrophysics Data System (ADS)
Hill, Steven C.; Pinnick, Ronald G.; Nachman, Paul; Chen, Gang; Chang, Richard K.; Mayo, Michael W.; Fernandez, Gilbert L.
1995-10-01
We have assembled an aerosol-fluorescence spectrum analyzer (AFS), which can measure the fluorescence spectra and elastic scattering of airborne particles as they flow through a laser beam. The aerosols traverse a scattering cell where they are illuminated with intense (50 kW/cm 2) light inside the cavity of an argon-ion laser operating at 488 nm. This AFS can obtain fluorescence spectra of individual dye-doped polystyrene microspheres as small as 0.5 mu m in diameter. The spectra obtained from microspheres doped with pink and green-yellow dyes are clearly different. We have also detected the fluorescence spectra of airborne particles (although not single particles) made from various
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zheng, Pei; Tong, Zhiqing; Bai, Bo, E-mail: baibochina@163.com
Porous hollow carbonaceous microspheres (PHCMs) fabricated from yeast cells by hydrothermal treatment have stimulated interest because of their outstanding chemical and physical properties. Herein, the functionalizations of PHCMs by further coating of α-Fe{sub 2}O{sub 3} nanoparticles onto the surface were carried out. The structure of resulted α-Fe{sub 2}O{sub 3}@PHCMs products were characterized by field emission scanning electron microscopy (FE-SEM), energy dispersive spectrometry (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), and BET specific surface area measurements (BET), respectively. Its promising application was evaluated by the Fenton-like degradation of fluorescent whitening agent-VBL from aqueous solutions. - Graphical abstract: In thismore » work, novel α-Fe{sub 2}O{sub 3}@porous hollow carbonaceous microspheres (α-Fe{sub 2}O{sub 3}@PHCMs) were synthesized through a combination of hydrothermal method and calcinations route and achieved excellent removal efficiency for fluorescent whitening Agent-VBL. - Highlights: • The hybrid α-Fe{sub 2}O{sub 3}@ porous hollow microspheres (PHCMs) were firstly fabricated. • The formation mechanism of α-Fe{sub 2}O{sub 3}@PHCMs microspheres was proposed and verified. • Dithizone played a key role in the synthesis of α-Fe{sub 2}O{sub 3}@PHCMs composites. • A favorable removal for the fluorescent whitening agent-VBL were achieved.« less
Multiplexed SNP genotyping using the Qbead™ system: a quantum dot-encoded microsphere-based assay
Xu, Hongxia; Sha, Michael Y.; Wong, Edith Y.; Uphoff, Janet; Xu, Yanzhang; Treadway, Joseph A.; Truong, Anh; O’Brien, Eamonn; Asquith, Steven; Stubbins, Michael; Spurr, Nigel K.; Lai, Eric H.; Mahoney, Walt
2003-01-01
We have developed a new method using the Qbead™ system for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The Qbead system employs fluorescent Qdot™ semiconductor nanocrystals, also known as quantum dots, to encode microspheres that subsequently can be used as a platform for multiplexed assays. By combining mixtures of quantum dots with distinct emission wavelengths and intensities, unique spectral ‘barcodes’ are created that enable the high levels of multiplexing required for complex genetic analyses. Here, we applied the Qbead system to SNP genotyping by encoding microspheres conjugated to allele-specific oligonucleotides. After hybridization of oligonucleotides to amplicons produced by multiplexed PCR of genomic DNA, individual microspheres are analyzed by flow cytometry and each SNP is distinguished by its unique spectral barcode. Using 10 model SNPs, we validated the Qbead system as an accurate and reliable technique for multiplexed SNP genotyping. By modifying the types of probes conjugated to microspheres, the Qbead system can easily be adapted to other assay chemistries for SNP genotyping as well as to other applications such as analysis of gene expression and protein–protein interactions. With its capability for high-throughput automation, the Qbead system has the potential to be a robust and cost-effective platform for a number of applications. PMID:12682378
Shih, Yen-Yu I.; Wang, Lin; De La Garza, Bryan H.; Li, Guang; Cull, Grant; Kiel, Jeffery W.; Duong, Timothy Q.
2013-01-01
Purpose The present study aimed to quantify retinal and choroidal blood flow (BF) during light, dark adaptation and flicker light stimulation using the microsphere technique. Materials and Methods Adult male Sprague–Dawley rats were anesthetized with isoflurane. Eyes were dark (Group I, n = 8), light (Group II, n = 8) adapted or stimulated with 10Hz flicker light (Group III, n = 10). Retinal and choroidal BF were measured by a previously established method, using a mixture of 8 μm yellow-green and 10 μm red fluorescent microspheres. The microspheres were counted ex vivo in the dissected retina and choroid and in the reference arterial blood under a fluorescent microscope. Results The choroidal BF was 64.8 ± 29 μl/min (mean ± SD) during dark adaptation, not significantly different from that during light adaptation (66.0 ± 17.8 μl/min). The retinal BF was 13.5 ± 3.2 μl/min during 10 Hz flickering light stimulation, significantly higher than that during dark adaptation in the control fellow eyes (9.9 ± 2.9 μl/min). The choroidal BF values were not statistically different between flicker stimulation and dark adaptation. Retinal BF was 11.6 ± 2.9 μl/min during light adaptation. Dark adaptation did not increase retinal BF (Group I, 8.2 ± 2.4 μl/min; Group II, 9.9 ± 2.9 μl/min). Conclusions These findings argue against a dark-induced or flicker-induced functional hyperemia in the choroid as a result of the demands of the outer retina. Retinal BF was not higher during dark adaptation. Our data support the conclusion that the inner retina has a higher energy demand in flicker conditions relative to dark. PMID:23317112
Shih, Yen-Yu I; Wang, Lin; De La Garza, Bryan H; Li, Guang; Cull, Grant; Kiel, Jeffery W; Duong, Timothy Q
2013-02-01
The present study aimed to quantify retinal and choroidal blood flow (BF) during light, dark adaptation and flicker light stimulation using the microsphere technique. Adult male Sprague-Dawley rats were anesthetized with isoflurane. Eyes were dark (Group I, n = 8), light (Group II, n = 8) adapted or stimulated with 10 Hz flicker light (Group III, n = 10). Retinal and choroidal BF were measured by a previously established method, using a mixture of 8 µm yellow-green and 10 µm red fluorescent microspheres. The microspheres were counted ex vivo in the dissected retina and choroid and in the reference arterial blood under a fluorescent microscope. The choroidal BF was 64.8 ± 29 µl/min (mean ± SD) during dark adaptation, not significantly different from that during light adaptation (66.0 ± 17.8 µl/min). The retinal BF was 13.5 ± 3.2 µl/min during 10 Hz flickering light stimulation, significantly higher than that during dark adaptation in the control fellow eyes (9.9 ± 2.9 µl/min). The choroidal BF values were not statistically different between flicker stimulation and dark adaptation. Retinal BF was 11.6 ± 2.9 µl/min during light adaptation. Dark adaptation did not increase retinal BF (Group I, 8.2 ± 2.4 µl/min; Group II, 9.9 ± 2.9 µl/min). These findings argue against a dark-induced or flicker-induced functional hyperemia in the choroid as a result of the demands of the outer retina. Retinal BF was not higher during dark adaptation. Our data support the conclusion that the inner retina has a higher energy demand in flicker conditions relative to dark.
Microsphere-Based Multiplex Analysis of DNA Methylation in Acute Myeloid Leukemia
Wertheim, Gerald B.W.; Smith, Catherine; Figueroa, Maria E.; Kalos, Michael; Bagg, Adam; Carroll, Martin; Master, Stephen R.
2015-01-01
Aberrant regulation of DNA methylation is characteristic of cancer cells and clearly influences phenotypes of various malignancies. Despite clear correlations between DNA methylation and patient outcome, tests that directly measure multiple-locus DNA methylation are typically expensive and technically challenging. Previous studies have demonstrated that the prognosis of patients with acute myeloid leukemia can be predicted by the DNA methylation pattern of 18 loci. We have developed a novel strategy, termed microsphere HpaII tiny fragment enrichment by ligation-mediated PCR (MELP), to simultaneously analyze the DNA methylation pattern at these loci using methylation-specific DNA digestion, fluorescently labeled microspheres, and branched DNA hybridization. The method uses techniques that are inexpensive and easily performed in a molecular laboratory. MELP accurately reflects the methylation levels at each locus analyzed and segregates patients with acute myeloid leukemia into prognostic subgroups. Our results demonstrate the usefulness of MELP as a platform for simultaneous evaluation of DNA methylation of multiple loci. PMID:24373919
Use of Protein A as the Primary Layer in Fluorescent Microsphere Technology.
1992-08-01
mAb) to the galactose-binding adherence lectin of Entamoeba histolytica were assessed for their abilities to bind protein A, using BlAcore. Of the six...permission of the Commander, U.S. Army Chemical Research, Development and Engineering Center (CRDEC), ATTN: SMCCR- SPS -T, Aberdeen Proving Ground, MD...14 6 USE OF PROTEIN A AS THE PRIMARY LAYER IN FLUORESCENT MICROSPHERE TECHNOLOGY 1. INTRODUCTION Entamoeba histolytica causes amebic colitis worldwide
NASA Astrophysics Data System (ADS)
Du Le, Vinh Nguyen; Patterson, Michael S.; Farrell, Thomas J.; Hayward, Joseph E.; Fang, Qiyin
2015-12-01
The ability to recover the intrinsic fluorescence of biological fluorophores is crucial to accurately identify the fluorophores and quantify their concentrations in the media. Although some studies have successfully retrieved the fluorescence spectral shape of known fluorophores, the techniques usually came with heavy computation costs and did not apply for strongly absorptive media, and the intrinsic fluorescence intensity and fluorophore concentration were not recovered. In this communication, an experimental approach was presented to recover intrinsic fluorescence and concentration of fluorescein in the presence of hemoglobin (Hb). The results indicated that the method was efficient in recovering the intrinsic fluorescence peak and fluorophore concentration with an error of 3% and 10%, respectively. The results also suggested that chromophores with irregular absorption spectra (e.g., Hb) have more profound effects on fluorescence spectral shape than chromophores with monotonic absorption and scattering spectra (e.g., black India ink and polystyrene microspheres).
Cai, Yanxue; Kang, Keren; Li, Qianru; Wang, Yu; He, Xiaowei
2018-05-07
A reliable lateral flow immunoassay (LFIA) based on a facile one-step synthesis of single microspheres in combining with immunochromatography technique was developed to establish a new point-of-care test (POCT) for the rapid and early detection of cardiac troponin I (cTnI), a kind of cardiac specific biomarker for acute myocardial infarction (AMI). The double layered microspheres with clear core-shell structures were produced using soap-free emulsion polymerization method with inexpensive compounds (styrene and acrylic acid). The synthetic process was simple, rapid and easy to control due to one-step synthesis without any complicated procedures. The microspheres are nanostructure with high surface area, which have numerous carboxyl groups on the out layer, resulting in high-efficiency coupling between the carrier and antibody via amide bond. Meanwhile, the red fluorescent dye, Nile-red (NR), was wrapped inside the microspheres to improve its stability, as well to reduce the background noise, because of its higher emission wavelength than interference from real plasma samples. The core-shell structures provided different functional areas to separate antibody and dyes, so the immunoassay has highly sensitive, wide working curves in the range of 0⁻40 ng/mL, low limits of detection (LOD) at 0.016 ng/mL, and limits of quantification (LOQ) at 0.087 ng/mL with coefficient of variations (CV) of 10%. This strategy suggested an outstanding platform for LFIA, with good reproducibility and stability to straightforwardly analyze the plasma samples without washing steps, thereby reducing the operating procedures for non-professionals and promoting detection efficiency. The whole detection process can be completed in less than 15 min. This novel immunoassay offers a reliable and favorable analytical result by detecting the real samples, indicating that it holds great potential as a new alternative for biomolecule detection in complex samples, for the early detection of cardiac specific biomarkers.
Diagnostic testing for Giardia infections.
Heyworth, Martin F
2014-03-01
The traditional method for diagnosing Giardia infections involves microscopic examination of faecal specimens for Giardia cysts. This method is subjective and relies on observer experience. From the 1980s onwards, objective techniques have been developed for diagnosing Giardia infections, and are superseding diagnostic techniques reliant on microscopy. Detection of Giardia antigen(s) by immunoassay is the basis of commercially available diagnostic kits. Various nucleic acid amplification techniques (NAATs) can demonstrate DNA of Giardia intestinalis, and have the potential to become standard approaches for diagnosing Giardia infections. Of such techniques, methods involving either fluorescent microspheres (Luminex) or isothermal amplification of DNA (loop-mediated isothermal amplification; LAMP) are especially promising.
Golden, J.P.; Verbarg, J.; Howell, P.B.; Shriver-Lake, L.C.; Ligler, F.S.
2012-01-01
A spinning magnetic trap (MagTrap) for automated sample processing was integrated with a microflow cytometer capable of simultaneously detecting multiple targets to provide an automated sample-to-answer diagnosis in 40 min. After target capture on fluorescently coded magnetic microspheres, the magnetic trap automatically concentrated the fluorescently coded microspheres, separated the captured target from the sample matrix, and exposed the bound target sequentially to biotinylated tracer molecules and streptavidin-labeled phycoerythrin. The concentrated microspheres were then hydrodynamically focused in a microflow cytometer capable of 4-color analysis (two wavelengths for microsphere identification, one for light scatter to discriminate single microspheres and one for phycoerythrin bound to the target). A three-fold decrease in sample preparation time and an improved detection limit, independent of target preconcentration, was demonstrated for detection of Escherichia coli 0157:H7 using the MagTrap as compared to manual processing. Simultaneous analysis of positive and negative controls, along with the assay reagents specific for the target, was used to obtain dose–response curves, demonstrating the potential for quantification of pathogen load in buffer and serum. PMID:22960010
Golden, J P; Verbarg, J; Howell, P B; Shriver-Lake, L C; Ligler, F S
2013-02-15
A spinning magnetic trap (MagTrap) for automated sample processing was integrated with a microflow cytometer capable of simultaneously detecting multiple targets to provide an automated sample-to-answer diagnosis in 40 min. After target capture on fluorescently coded magnetic microspheres, the magnetic trap automatically concentrated the fluorescently coded microspheres, separated the captured target from the sample matrix, and exposed the bound target sequentially to biotinylated tracer molecules and streptavidin-labeled phycoerythrin. The concentrated microspheres were then hydrodynamically focused in a microflow cytometer capable of 4-color analysis (two wavelengths for microsphere identification, one for light scatter to discriminate single microspheres and one for phycoerythrin bound to the target). A three-fold decrease in sample preparation time and an improved detection limit, independent of target preconcentration, was demonstrated for detection of Escherichia coli 0157:H7 using the MagTrap as compared to manual processing. Simultaneous analysis of positive and negative controls, along with the assay reagents specific for the target, was used to obtain dose-response curves, demonstrating the potential for quantification of pathogen load in buffer and serum. Published by Elsevier B.V.
Characterization of buoyant fluorescent particles for field observations of water flows.
Tauro, Flavia; Aureli, Matteo; Porfiri, Maurizio; Grimaldi, Salvatore
2010-01-01
In this paper, the feasibility of off-the-shelf buoyant fluorescent microspheres as particle tracers in turbid water flows is investigated. Microspheres' fluorescence intensity is experimentally measured and detected in placid aqueous suspensions of increasing concentrations of clay to simulate typical conditions occurring in natural drainage networks. Experiments are conducted in a broad range of clay concentrations and particle immersion depths by using photoconductive cells and image-based sensing technologies. Results obtained with both methodologies exhibit comparable trends and show that the considered particles are fairly detectable in critically turbid water flows. Further information on performance and integration of the studied microspheres in low-cost measurement instrumentation for field observations is obtained through experiments conducted in a custom built miniature water channel. This experimental characterization provides a first assessment of the feasibility of commercially available buoyant fluorescent beads in the analysis of high turbidity surface water flows. The proposed technology may serve as a minimally invasive sensing system for hazardous events, such as pollutant diffusion in natural streams and flash flooding due to extreme rainfall.
Laser velocimetry with fluorescent dye-doped polystyrene microspheres.
Lowe, K Todd; Maisto, Pietro; Byun, Gwibo; Simpson, Roger L; Verkamp, Max; Danehy, Paul M; Tiemsin, Pacita I; Wohl, Christopher J
2013-04-15
Simultaneous Mie scattering and laser-induced fluorescence (LIF) signals are obtained from individual polystyrene latex microspheres dispersed in an air flow. Microspheres less than 1 μm mean diameter were doped with two organic fluorescent dyes, Rhodamine B (RhB) and dichlorofluorescein (DCF), intended either to provide improved particle-based flow velocimetry in the vicinity of surfaces or to provide scalar flow information (e.g., marking one of two fluid streams). Both dyes exhibit measureable fluorescence signals that are on the order of 10(-3) to 10(-4) times weaker than the simultaneously measured Mie signals. It is determined that at the conditions measured, 95.5% of RhB LIF signals and 32.2% of DCF signals provide valid laser-Doppler velocimetry measurements compared with the Mie scattering validation rate with 6.5 W of 532 nm excitation, while RhB excited with 1.0 W incident laser power still exhibits 95.4% valid velocimetry signals from the LIF channel. The results suggest that the method is applicable to wind tunnel measurements near walls where laser flare can be a limiting factor and monodisperse particles are essential.
Sousa, Susana; Cardoso, Luís; Reed, Steven G.; Reis, Alexandre B.; Martins-Filho, Olindo A.; Silvestre, Ricardo; Cordeiro da Silva, Anabela
2013-01-01
Background An accurate diagnosis is essential for the control of infectious diseases. In the search for effective and efficient tests, biosensors have increasingly been exploited for the development of new and highly sensitive diagnostic methods. Here, we describe a new fluorescent based immunosensor comprising magnetic polymer microspheres coated with recombinant antigens to improve the detection of specific antibodies generated during an infectious disease. As a challenging model, we used canine leishmaniasis due to the unsatisfactory sensitivity associated with the detection of infection in asymptomatic animals where the levels of pathogen-specific antibodies are scarce. Methodology Ni-NTA magnetic microspheres with 1,7 µm and 8,07 µm were coated with the Leishmania recombinant proteins LicTXNPx and rK39, respectively. A mixture of equal proportions of both recombinant protein-coated microspheres was used to recognize and specifically bind anti-rK39 and anti-LicTNXPx antibodies present in serum samples of infected dogs. The microspheres were recovered by magnetic separation and the percentage of fluorescent positive microspheres was quantified by flow cytometry. Principal Findings A clinical evaluation carried out with 129 dog serum samples using the antigen combination demonstrated a sensitivity of 98,8% with a specificity of 94,4%. rK39 antigen alone demonstrated a higher sensitivity for symptomatic dogs (96,9%), while LicTXNPx antigen showed a higher sensitivity for asymptomatic (94,4%). Conclusions Overall, our results demonstrated the potential of a magnetic microsphere associated flow cytometry methodology as a viable tool for highly sensitive laboratorial serodiagnosis of both clinical and subclinical forms of canine leishmaniasis. PMID:23991232
Lee, H; Nguyen, T T; Kim, M; Jeong, J-H; Park, J-B
2018-05-31
Quercetin has been reported to exert many beneficial effects on the protection against various diseases, such as diabetes, cancer, and inflammation. The aim of this study is to evaluate the potential osteogenic differentiation ability of mesenchymal stem cells in the presence of quercetin. Quercetin-loaded poly(lactic-co-glycolic acid) microspheres were prepared using an electrospraying technique. Characterization of the microspheres was evaluated with a scanning electron microscope and release profile. Three-dimensional cell spheroids were fabricated using silicon elastomer-based concave microwells. Qualitative results of cellular viability were seen under a confocal microscope, and quantitative cellular viability was evaluated using the Cell Counting Kit-8 assay. The alkaline phosphatase activity and Alizarin Red S staining were performed. A quantitative real-time polymerase chain reaction and a western blot analysis were performed. Spheroids were well formed irrespective of quercetin concentration. Most of the cells in spheroids emitted green fluorescence, and the morphology was round without significant changes. The application of quercetin-loaded microspheres produced a significant increase in the alkaline phosphatase activity. The real-time polymerase chain reaction results showed a significant increase in Runx2, and western blot results showed higher expression of Runx2 protein expression. Biodegradable microspheres loaded with quercetin produced prolonged release profiles with increased mineralization. Microspheres loaded with quercetin can be used for the enhancement of osteoblastic differentiation in cell therapy. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Kolin, David L.; Ronis, David; Wiseman, Paul W.
2006-01-01
We present the theory and application of reciprocal space image correlation spectroscopy (kICS). This technique measures the number density, diffusion coefficient, and velocity of fluorescently labeled macromolecules in a cell membrane imaged on a confocal, two-photon, or total internal reflection fluorescence microscope. In contrast to r-space correlation techniques, we show kICS can recover accurate dynamics even in the presence of complex fluorophore photobleaching and/or “blinking”. Furthermore, these quantities can be calculated without nonlinear curve fitting, or any knowledge of the beam radius of the exciting laser. The number densities calculated by kICS are less sensitive to spatial inhomogeneity of the fluorophore distribution than densities measured using image correlation spectroscopy. We use simulations as a proof-of-principle to show that number densities and transport coefficients can be extracted using this technique. We present calibration measurements with fluorescent microspheres imaged on a confocal microscope, which recover Stokes-Einstein diffusion coefficients, and flow velocities that agree with single particle tracking measurements. We also show the application of kICS to measurements of the transport dynamics of α5-integrin/enhanced green fluorescent protein constructs in a transfected CHO cell imaged on a total internal reflection fluorescence microscope using charge-coupled device area detection. PMID:16861272
Fiber-optic array using molecularly imprinted microspheres for antibiotic analysis.
Carrasco, Sergio; Benito-Peña, Elena; Walt, David R; Moreno-Bondi, María C
2015-05-01
In this article we describe a new class of high-density optical microarrays based on molecularly imprinted microsphere sensors that directly incorporate specific recognition capabilities to detect enrofloxacin (ENRO), an antibiotic widely used for both human and veterinary applications. This approach involves the preparation of highly cross-linked polymer microspheres by thermal precipitation-polymerization in the presence and absence of the target analyte ENRO to generate either molecularly imprinted (MIP) or non-imprinted polymer (NIP) microspheres, respectively. Each polymer type of tailor-made microsphere is fluorescently encoded with either coumarin-30 or tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(ii) dichloride [Ru(dip) 3 ]Cl 2 to enable the microspheres to be distinguished. The new MIP-based sensing platform utilizes an optical fiber bundle containing approximately 50 000 individual 3.1 μm diameter fibers that are chemically etched to create microwells in which MIP and NIP microspheres can be deposited and imaged using an epi-fluorescence microscope. The method enables multiplexed detection by independently addressing both types of beads through their separate light channels. The unique response to the presence of ENRO is manifested on the basis of a competitive immunoassay. A red-fluorescent dye-tagged ENRO, labeled with BODIPY® TR Cadaverine, competes with ENRO for specific binding sites. The developed immuno-like assay displayed a limit of detection (LOD) of 0.04 μM (10% binding inhibition) and a dynamic range of 0.29-21.54 μM (20-80% binding inhibition). The selectivity of the assay was evaluated by measuring the cross-reactivity of other fluoroquinolones (ciprofloxacin, norfloxacin, danofloxacin, and flumequine) and non-related antibiotics (penicillin G and doxycycline). This work demonstrates, for the first time, the applicability of MIPs, as an alternative to biomolecule receptors, for the development of multiplexed detection fiber-optic microarrays paving the way for a new generation of biomimetic sensors.
Digital Analysis and Sorting of Fluorescence Lifetime by Flow Cytometry
Houston, Jessica P.; Naivar, Mark A.; Freyer, James P.
2010-01-01
Frequency-domain flow cytometry techniques are combined with modifications to the digital signal processing capabilities of the Open Reconfigurable Cytometric Acquisition System (ORCAS) to analyze fluorescence decay lifetimes and control sorting. Real-time fluorescence lifetime analysis is accomplished by rapidly digitizing correlated, radiofrequency modulated detector signals, implementing Fourier analysis programming with ORCAS’ digital signal processor (DSP) and converting the processed data into standard cytometric list mode data. To systematically test the capabilities of the ORCAS 50 MS/sec analog-to-digital converter (ADC) and our DSP programming, an error analysis was performed using simulated light scatter and fluorescence waveforms (0.5–25 ns simulated lifetime), pulse widths ranging from 2 to 15 µs, and modulation frequencies from 2.5 to 16.667 MHz. The standard deviations of digitally acquired lifetime values ranged from 0.112 to >2 ns, corresponding to errors in actual phase shifts from 0.0142° to 1.6°. The lowest coefficients of variation (<1%) were found for 10-MHz modulated waveforms having pulse widths of 6 µs and simulated lifetimes of 4 ns. Direct comparison of the digital analysis system to a previous analog phase-sensitive flow cytometer demonstrated similar precision and accuracy on measurements of a range of fluorescent microspheres, unstained cells and cells stained with three common fluorophores. Sorting based on fluorescence lifetime was accomplished by adding analog outputs to ORCAS and interfacing with a commercial cell sorter with a radiofrequency modulated solid-state laser. Two populations of fluorescent microspheres with overlapping fluorescence intensities but different lifetimes (2 and 7 ns) were separated to ~98% purity. Overall, the digital signal acquisition and processing methods we introduce present a simple yet robust approach to phase-sensitive measurements in flow cytometry. The ability to simply and inexpensively implement this system on a commercial flow sorter will both allow better dissemination of this technology and better exploit the traditionally underutilized parameter of fluorescence lifetime. PMID:20662090
Resolution enhancement of 2-photon microscopy using high-refractive index microspheres
NASA Astrophysics Data System (ADS)
Tehrani, Kayvan Forouhesh; Darafsheh, Arash; Phang, Sendy; Mortensen, Luke J.
2018-02-01
Intravital microscopy using multiphoton processes is the standard tool for deep tissue imaging inside of biological specimens. Usually, near-infrared and infrared light is used to excite the sample, which enables imaging several mean free path inside a scattering tissues. Using longer wavelengths, however, increases the width of the effective multiphoton Point Spread Function (PSF). Many features inside of cells and tissues are smaller than the diffraction limit, and therefore not possible to distinguish using a large PSF. Microscopy using high refractive index microspheres has shown promise to increase the numerical aperture of an imaging system and enhance the resolution. It has been shown that microspheres can image features λ/7 using single photon process fluorescence. In this work, we investigate resolution enhancement for Second Harmonic Generation (SHG) and 2-photon fluorescence microscopy. We used Barium Titanate glass microspheres with diameters ˜20-30 μm and refractive index ˜1.9-2.1. We show microsphere-assisted SHG imaging in bone collagen fibers. Since bone is a very dense tissue constructed of bundles of collagen fibers, it is nontrivial to image individual fibers. We placed microspheres on a dense area of the mouse cranial bone, and achieved imaging of individual fibers. We found that microsphere assisted SHG imaging resolves features of the bone fibers that are not readily visible in conventional SHG imaging. We extended this work to 2-photon microscopy of mitochondria in mouse soleus muscle, and with the help of microsphere resolving power, we were able to trace individual mitochondrion from their ensemble.
Optofluidic microscope with 3D spatial resolution.
Vig, Asger Laurburg; Marie, Rodolphe; Jensen, Eric; Kristensen, Anders
2010-03-01
This paper reports on-chip based optical detection with three-dimensional spatial resolution by integration of an optofluidic microscope (OFM) in a microfluidic pinched flow fractionation (PFF) separation device. This setup also enables on-chip particle image velocimetry (PIV). The position in the plane perpendicular to the flow direction and the velocity along the flow direction of separated fluorescent labeled polystyrene microspheres with diameters of 1 microm , 2.1 microm , 3 microm and 4 microm is determined by the OFM. These results are bench marked against those obtained with a PFF device using conventional fluorescence microscope readout. The size separated microspheres are detected by OFM with an accuracy of
Development and validation of a fluorescent microsphere immunoassay for soluble CD30 testing.
Pavlov, Igor; Martins, Thomas B; Delgado, Julio C
2009-09-01
Testing for soluble CD30 (sCD30), an indicator of Th2 immune response, is a useful prognostic marker in solid organ transplantation, lymphoproliferative disorders, autoimmunity, and various parasitic diseases. In this study we report the development and validation of a fluorescent microsphere immunoassay for the detection of sCD30 in serum, plasma, and culture supernatants. The dynamic range of this assay is 1 to 400 ng/ml, and the rate of recovery of various concentrations of recombinant sCD30 ranges from 97 to 116% (average recovery, 105%). The test showed a high degree of precision in both intra-assay and interassay studies (coefficients of variation, as high as 7% and 8%, respectively), with a sensitivity of 1 ng/ml. The normal reference range calculated for a cohort of 151 healthy individuals was 1 to 29 ng/ml. The clinical usefulness of the sCD30 fluorescent microsphere immunoassay was demonstrated by showing that levels of sCD30 have a positive correlation with specimens containing high titers of anti-double-stranded DNA antibodies and high titers of immunoglobulin G against Leishmania species. Given the multiplexing potential of the sCD30 fluorescent microsphere immunoassay reported in this study, it is expected that testing of sCD30 concentrations along with those of other cytokines will become an important diagnostic tool for selected immunological and inflammatory diseases where Th2-type cytokine responses have been reported.
Development and Validation of a Fluorescent Microsphere Immunoassay for Soluble CD30 Testing▿
Pavlov, Igor; Martins, Thomas B.; Delgado, Julio C.
2009-01-01
Testing for soluble CD30 (sCD30), an indicator of Th2 immune response, is a useful prognostic marker in solid organ transplantation, lymphoproliferative disorders, autoimmunity, and various parasitic diseases. In this study we report the development and validation of a fluorescent microsphere immunoassay for the detection of sCD30 in serum, plasma, and culture supernatants. The dynamic range of this assay is 1 to 400 ng/ml, and the rate of recovery of various concentrations of recombinant sCD30 ranges from 97 to 116% (average recovery, 105%). The test showed a high degree of precision in both intra-assay and interassay studies (coefficients of variation, as high as 7% and 8%, respectively), with a sensitivity of 1 ng/ml. The normal reference range calculated for a cohort of 151 healthy individuals was 1 to 29 ng/ml. The clinical usefulness of the sCD30 fluorescent microsphere immunoassay was demonstrated by showing that levels of sCD30 have a positive correlation with specimens containing high titers of anti-double-stranded DNA antibodies and high titers of immunoglobulin G against Leishmania species. Given the multiplexing potential of the sCD30 fluorescent microsphere immunoassay reported in this study, it is expected that testing of sCD30 concentrations along with those of other cytokines will become an important diagnostic tool for selected immunological and inflammatory diseases where Th2-type cytokine responses have been reported. PMID:19605595
Pinhole shifting lifetime imaging microscopy
Ramshesh, Venkat K.; Lemasters, John J.
2009-01-01
Lifetime imaging microscopy is a powerful tool to probe biological phenomena independent of luminescence intensity and fluorophore concentration. We describe time-resolved imaging of long-lifetime luminescence with an unmodified commercial laser scanning confocal/multiphoton microscope. The principle of the measurement is displacement of the detection pinhole to collect delayed luminescence from a position lagging the rasting laser beam. As proof of principle, luminescence from microspheres containing europium (Eu3+), a red emitting probe, was compared to that of short-lifetime green-fluorescing microspheres and/or fluorescein and rhodamine in solution. Using 720-nm two-photon excitation and a pinhole diameter of 1 Airy unit, the short-lifetime fluorescence of fluorescein, rhodamine and green microspheres disappeared much more rapidly than the long-lifetime phosphorescence of Eu3+ microspheres as the pinhole was repositioned in the lagging direction. In contrast, repositioning of the pinhole in the leading and orthogonal directions caused equal loss of short- and long-lifetime luminescence. From measurements at different lag pinhole positions, a lifetime of 270 μs was estimated for the Eu3+ microspheres, consistent with independent measurements. This simple adaptation is the basis for quantitative 3-D lifetime imaging microscopy. PMID:19123648
Screening unlabeled DNA targets with randomly ordered fiber-optic gene arrays.
Steemers, F J; Ferguson, J A; Walt, D R
2000-01-01
We have developed a randomly ordered fiber-optic gene array for rapid, parallel detection of unlabeled DNA targets with surface immobilized molecular beacons (MB) that undergo a conformational change accompanied by a fluorescence change in the presence of a complementary DNA target. Microarrays are prepared by randomly distributing MB-functionalized 3-microm diameter microspheres in an array of wells etched in a 500-microm diameter optical imaging fiber. Using several MBs, each designed to recognize a different target, we demonstrate the selective detection of genomic cystic fibrosis related targets. Positional registration and fluorescence response monitoring of the microspheres was performed using an optical encoding scheme and an imaging fluorescence microscope system.
NASA Astrophysics Data System (ADS)
Fan, Wenhui; Li, Qing; Hu, Liang; Yan, Siqi; Wen, Wanxin; Chai, Zhifang; Liu, Hanzhou
2017-01-01
To simply and multitudinously synthesize hollow microspheres in a pure system is important for relevant research and application. Here, a simple and novel one-pot synthetic strategy to prepare polystyrene (PS) hollow microspheres via irradiation-assisted free-radical polymerizing and self-assembly (IFPS) approach under γ-ray irradiation with no additives introduced into the system is presented. And PS/2,5-Diphenyloxazole (PPO) fluorescent microspheres have been prepared successfully by IFPS reaction, which can be used as scintillators for the detection of ionizing radiation. A linear relationship between emitted luminescence and dose-activity in water is obtained, which suggests that composite microspheres could be used as liquid scintillation in specific environment.
Ludwig, Susann K J; Zhu, Hongying; Phillips, Stephen; Shiledar, Ashutosh; Feng, Steve; Tseng, Derek; van Ginkel, Leendert A; Nielen, Michel W F; Ozcan, Aydogan
2014-11-01
Current contaminant and residue monitoring throughout the food chain is based on sampling, transport, administration, and analysis in specialized control laboratories. This is a highly inefficient and costly process since typically more than 99% of the samples are found to be compliant. On-site simplified prescreening may provide a scenario in which only samples that are suspect are transported and further processed. Such a prescreening can be performed using a small attachment on a cellphone. To this end, a cellphone-based imaging platform for a microsphere fluorescence immunoassay that detects the presence of anti-recombinant bovine somatotropin (rbST) antibodies in milk extracts was developed. RbST administration to cows increases their milk production, but is illegal in the EU and a public health concern in the USA. The cellphone monitors the presence of anti-rbST antibodies (rbST biomarker), which are endogenously produced upon administration of rbST and excreted in milk. The rbST biomarker present in milk extracts was captured by rbST covalently coupled to paramagnetic microspheres and labeled by quantum dot (QD)-coupled detection antibodies. The emitted fluorescence light from these captured QDs was then imaged using the cellphone camera. Additionally, a dark-field image was taken in which all microspheres present were visible. The fluorescence and dark-field microimages were analyzed using a custom-developed Android application running on the same cellphone. With this setup, the microsphere fluorescence immunoassay and cellphone-based detection were successfully applied to milk sample extracts from rbST-treated and untreated cows. An 80% true-positive rate and 95% true-negative rate were achieved using this setup. Next, the cellphone-based detection platform was benchmarked against a newly developed planar imaging array alternative and found to be equally performing versus the much more sophisticated alternative. Using cellphone-based on-site analysis in future residue monitoring can limit the number of samples for laboratory analysis already at an early stage. Therewith, the entire monitoring process can become much more efficient and economical.
Wang, Qiang-Song; Zhu, Xiao-Ning; Jiang, Heng-Li; Wang, Gui-Fang; Cui, Yuan-Lu
2015-01-01
Zuojin Pill (ZJP), a traditional Chinese medicine formula, consists of Coptis chinensis Franch. and Evodia rutaecarpa (Juss.) Benth. in a ratio of 6:1 (w/w) and was first recorded in “Danxi’s experiential therapy” for treating gastrointestinal disorders in the 15th century. However, the poor solubility of alkaloids from ZJP restricted the protective effect in treating gastritis and gastric ulcer. The aim of the study was to investigate the protective mechanism of mucoadhesive microspheres loaded with alkaloids from C. chinensis Franch. and E. rutaecarpa (Juss.) Benth. on ethanol-induced acute gastric mucosal injury in rats. Surface morphology, particle size, drug loading, encapsulation efficiency, in vitro drug release, mucoadhesiveness, and fluorescent imaging of the microspheres in gastrointestinal tract were studied. The results showed that the mucoadhesive microspheres loaded with alkaloids could sustain the release of drugs beyond 12 hours and had gastric mucoadhesive property with 82.63% retention rate in vitro. The fluorescence tracer indicated high retention of mucoadhesive microspheres within 12 hours in vivo. The mucoadhesive microspheres loaded with alkaloids could reduce the gastric injury by decreasing the mucosal lesion index, increasing the percentage of inhibition and increasing the amount of mucus in the gastric mucosa in an ethanol-induced gastric mucosal injury rat model. Moreover, the mucoadhesive microspheres loaded with alkaloids reduce the inflammatory response by decreasing the levels of tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), downregulating the mRNA expression of inducible nitric oxide synthase, TNF-α, and IL-1β in gastric mucosa. All the results indicate that mucoadhesive microspheres loaded with alkaloids could not only increase the residence time of alkaloids in rat stomach, but also exert gastroprotective effects through reducing the inflammatory response on ethanol-induced gastric mucosal damage. Thus, these microspheres could be developed as a potential controlled release drug for treatment of gastric ulcer. PMID:26640368
Control of silk microsphere formation using polyethylene glycol (PEG).
Wu, Jianbing; Zheng, Zhaozhu; Li, Gang; Kaplan, David L; Wang, Xiaoqin
2016-07-15
A one step, rapid method to prepare silk microspheres was developed, with particle size controlled by the addition of polyethylene glycol (PEG). PEG molecular weight (4.0K-20.0KDa) and concentration (20-50wt%), as well as silk concentration (5-20wt%), were key factors that determined particle sizes varying in a range of 1-100μm. Addition of methanol to the PEG-silk combinations increased the content of crystalline β-sheet in the silk microspheres. To track the distribution and degradation of silk microspheres in vivo, 3-mercaptopropionic acid (MPA)-coated CdTe quantum dots (QDs) were physically entrapped in the silk microspheres. QDs tightly bound to the β-sheet domains of silk via hydrophobic interactions, with over 96% of the loaded QDs remaining in the silk microspheres after exhaustive extraction. The fluorescence of QDs-incorporated silk microspheres less stable in cell culture medium than in phosphate buffer solution (PBS) and water. After subcutaneous injection in mice, microspheres prepared from 20% silk (approx. 30μm diameter particles) still fluoresced at 24h, while those prepared from 8% silk (approx. 4μm diameter particles) and free QDs were not detectable, reflecting the QDs quenching and particle size effect on microsphere clearance in vivo. The larger microspheres were more resistant to cell internalization and degradation. Since PEG is an FDA-approved polymer, and silk is FDA approved for some medical devices, the methods developed in the present study will be useful in a variety of biomedical applications where simple, rapid and scalable preparation of silk microspheres is required. The work is of significance to the biomaterial and controlled release society because it provides a new option for fabricating silk microspheres in one simple step of mixing silk and polyethylene glycol (PEG), with the size and properties of microspheres controllable by PEG molecular weight as well as PEG and silk concentrations. Although fabrication of silk microspheres have been reported previously using spray-drying, liposome-templating, polyvinyl alcohol (PVA) emulsification, etc., applications were hindered due to harsh conditions (temperature, solvents, etc.) and complicated procedures used as well as low yield and less controllable particle size (usually <10μm). Since PEG is an FDA-approved polymer, and silk is FDA approved for some medical devices, the methods developed in the present study will be useful in a variety of biomedical applications where simple, rapid and scalable preparation of silk microspheres is required. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
Yuan, Pingfan; Ma, Qiang; Meng, Rizeng; Wang, Chao; Dou, Wenchao; Wang, Guannan; Su, Xingguang
2009-05-01
Semiconductor nanocrystals (or quantum dots, QDs) have the potential to overcome some of the limitations encountered by traditional fluorophores in fluorescence labeling applications. The unique spectroscopic properties of QDs make them hold immense promise as versatile labels for biological applications. In this work, we employ the layer-by-layer (LbL) method for the construction of bio-functional multicolor QD-encoded microspheres. Polystyrene microspheres with diameter of 3 microm were used as templates for the deposition of different sized CdTe QDs/polyelectrolyte multilayers. Two different antigens, Chicken newcastle disease (CND) antigen and goat pox virus (GPV) antigen, were conjugated to two kinds of biofunctional multicolor microspheres with different optical encoding. The multicolor microspheres can capture corresponding antibodies labeled with QDs, QDs-CND antibody and QDs-GPV antibody in the fluoroimmunoassays. The microspheres can be distinguished from each other based on their optical encoding.
Jogiya, Roy; Makowski, Markus; Phinikaridou, Alkystsis; Patel, Ashish S; Jansen, Christian; Zarinabad, Niloufar; Chiribiri, Amedeo; Botnar, Rene; Nagel, Eike; Kozerke, Sebastian; Plein, Sven
2013-07-21
Dynamic first pass contrast-enhanced myocardial perfusion is the standard CMR method for the estimation of myocardial blood flow (MBF) and MBF reserve in man, but it is challenging in rodents because of the high temporal and spatial resolution requirements. Hyperemic first pass myocardial perfusion CMR during vasodilator stress in mice has not been reported. Five C57BL/6 J mice were scanned on a clinical 3.0 Tesla Achieva system (Philips Healthcare, Netherlands). Vasodilator stress was induced via a tail vein catheter with an injection of dipyridamole. Dynamic contrast-enhanced perfusion imaging (Gadobutrol 0.1 mmol/kg) was based on a saturation recovery spoiled gradient echo method with 10-fold k-space and time domain undersampling (k-t PCA). One week later the mice underwent repeat anaesthesia and LV injections of fluorescent microspheres at rest and at stress. Microspheres were analysed using confocal microscopy and fluorescence-activated cell sorting. Mean MBF at rest measured by Fermi-function constrained deconvolution was 4.1 ± 0.5 ml/g/min and increased to 9.6 ± 2.5 ml/g/min during dipyridamole stress (P = 0.005). The myocardial perfusion reserve was 2.4 ± 0.54. The mean count ratio of stress to rest microspheres was 2.4 ± 0.51 using confocal microscopy and 2.6 ± 0.46 using fluorescence. There was good agreement between cardiovascular magnetic resonance CMR and microspheres with no significant difference (P = 0.84). First-pass myocardial stress perfusion CMR in a mouse model is feasible at 3 Tesla. Rest and stress MBF values were consistent with existing literature and perfusion reserve correlated closely to microsphere analysis. Data were acquired on a 3 Tesla scanner using an approach similar to clinical acquisition protocols, potentially facilitating translation of imaging findings between rodent and human studies.
Biological applications of confocal fluorescence polarization microscopy
NASA Astrophysics Data System (ADS)
Bigelow, Chad E.
Fluorescence polarization microscopy is a powerful modality capable of sensing changes in the physical properties and local environment of fluorophores. In this thesis we present new applications for the technique in cancer diagnosis and treatment and explore the limits of the modality in scattering media. We describe modifications to our custom-built confocal fluorescence microscope that enable dual-color imaging, optical fiber-based confocal spectroscopy and fluorescence polarization imaging. Experiments are presented that indicate the performance of the instrument for all three modalities. The limits of confocal fluorescence polarization imaging in scattering media are explored and the microscope parameters necessary for accurate polarization images in this regime are determined. A Monte Carlo routine is developed to model the effect of scattering on images. Included in it are routines to track the polarization state of light using the Mueller-Stokes formalism and a model for fluorescence generation that includes sampling the excitation light polarization ellipse, Brownian motion of excited-state fluorophores in solution, and dipole fluorophore emission. Results from this model are compared to experiments performed on a fluorophore-embedded polymer rod in a turbid medium consisting of polystyrene microspheres in aqueous suspension. We demonstrate the utility of the fluorescence polarization imaging technique for removal of contaminating autofluorescence and for imaging photodynamic therapy drugs in cell monolayers. Images of cells expressing green fluorescent protein are extracted from contaminating fluorescein emission. The distribution of meta-tetrahydroxypheny1chlorin in an EMT6 cell monolayer is also presented. A new technique for imaging enzyme activity is presented that is based on observing changes in the anisotropy of fluorescently-labeled substrates. Proof-of-principle studies are performed in a model system consisting of fluorescently labeled bovine serum albumin attached to sepharose beads. The action of trypsin and proteinase K on the albumin is monitored to demonstrate validity of the technique. Images of the processing of the albumin in J774 murine macrophages are also presented indicating large intercellular differences in enzyme activity. Future directions for the technique are also presented, including the design of enzyme probes specific for prostate specific antigen based on fluorescently-labeled dendrimers. A technique for enzyme imaging based on extracellular autofluorescence is also proposed.
Nano-functionalization of protein microspheres
NASA Astrophysics Data System (ADS)
Yoon, Sungkwon; Nichols, William T.
2014-08-01
Protein microspheres are promising building blocks for the assembly of complex functional materials. Here we demonstrate a set of three techniques that add functionality to the surface of protein microspheres. In the first technique, a positive surface charge on the protein spheres is deposited by electrostatic adsorption. Negatively charged silica and gold nanoparticle colloids can then electrostatically bind reversibly to the microsphere surface. In the second technique, nanoparticles are covalently anchored to the protein shell using a simple one-pot process. The strong covalent bond between sulfur groups in cysteine in the protein shell irreversibly binds to the gold nanoparticles. In the third technique, surface morphology of the protein microsphere is tuned through hydrodynamic instability at the water-oil interface. This is accomplished through the degree of solubility of the oil phase in water. Taken together these three techniques form a platform to create nano-functionalized protein microspheres, which can then be used as building blocks for the assembly of more complex macroscopic materials.
Chowdhury, Shwetadwip; Eldridge, Will J.; Wax, Adam; Izatt, Joseph A.
2017-01-01
Sub-diffraction resolution imaging has played a pivotal role in biological research by visualizing key, but previously unresolvable, sub-cellular structures. Unfortunately, applications of far-field sub-diffraction resolution are currently divided between fluorescent and coherent-diffraction regimes, and a multimodal sub-diffraction technique that bridges this gap has not yet been demonstrated. Here we report that structured illumination (SI) allows multimodal sub-diffraction imaging of both coherent quantitative-phase (QP) and fluorescence. Due to SI’s conventionally fluorescent applications, we first demonstrate the principle of SI-enabled three-dimensional (3D) QP sub-diffraction imaging with calibration microspheres. Image analysis confirmed enhanced lateral and axial resolutions over diffraction-limited QP imaging, and established striking parallels between coherent SI and conventional optical diffraction tomography. We next introduce an optical system utilizing SI to achieve 3D sub-diffraction, multimodal QP/fluorescent visualization of A549 biological cells fluorescently tagged for F-actin. Our results suggest that SI has a unique utility in studying biological phenomena with significant molecular, biophysical, and biochemical components. PMID:28663887
Metge, D.W.; Harvey, R.W.; Anders, R.; Rosenberry, D.O.; Seymour, D.; Jasperse, James
2007-01-01
Carboxylated microspheres were employed as surrogates to assess the transport potential of Cryptosporidium parvumoocysts during forced- and natural-gradient tests conducted in July and October 2004. The tests involved poorly-sorted, near-surface sediments where groundwater is pumped from an alluvial aquifer underlying the Russian River, Sonoma County, CA. In an off channel infiltration basin and within the river, a mixture (2-, 3-, and 5- ??m diameters) of fluorescently-labeled carboxylated microspheres and bromide tracers were used in two injection and recovery test to assess sediment removal efficiency for the microspheres. Bottom sediments varied considerably in their filtration efficiency for Cryptosporidium.
NASA Technical Reports Server (NTRS)
Rhim, Won-Kyu; Hyson, Michael T.; Chung, Sang-Kun; Colvin, Michael S.; Chang, Manchium
1989-01-01
Combination of advanced techniques yields uniform particles for biomedical applications. Process combines ink-jet and irradiation/freeze-polymerization techniques to make polymeric microspheres of uniform size in diameters from 100 to 400 micrometer. Microspheres used in chromatography, cell sorting, cell labeling, and manufacture of pharmaceutical materials.
High-density fiber-optic DNA random microsphere array.
Ferguson, J A; Steemers, F J; Walt, D R
2000-11-15
A high-density fiber-optic DNA microarray sensor was developed to monitor multiple DNA sequences in parallel. Microarrays were prepared by randomly distributing DNA probe-functionalized 3.1-microm-diameter microspheres in an array of wells etched in a 500-microm-diameter optical imaging fiber. Registration of the microspheres was performed using an optical encoding scheme and a custom-built imaging system. Hybridization was visualized using fluorescent-labeled DNA targets with a detection limit of 10 fM. Hybridization times of seconds are required for nanomolar target concentrations, and analysis is performed in minutes.
NASA Astrophysics Data System (ADS)
Nguyen, Dam Thuy Trang; Pelissier, Aurélien; Montes, Kevin; Tong, Quang Cong; Ngo, Hoang Minh; Ledoux-Rak, Isabelle; Lai, Ngoc Diep
2016-04-01
We have precisely positioned and embedded a single gold nanoparticle (Au NP) into a desired polymeric photonic structure (PS) using a simple and low-cost technique called low one-photon absorption direct laser writing (LOPA DLW), with a two-step process: identification and fabrication. First, the position of the Au NP was identified with a precision of 20 nm by using DLW technique with ultralow excitation laser power (μW). This power did not induce the polymerization of the photoresist (SU8) due to its low absorption at the excitation wavelength (532 nm). Then, the structure containing the NP was fabricated by using the same DLW system with high excitation power (mW). Different 2D photonic structures have been fabricated, which contain a single Au NP at desired position. In particular, we obtained a microsphere instead of a micropillar at the position of the Au NP. The formation of such microsphere was explained by the thermal effect of the Au NP at the wavelength of 532 nm, which induced thermal polymerization of surrounding photoresist. The effect of the post-exposure bake on the quality of structures was taken into account, revealing a more efficient fabrication way by exploiting the local thermal effect of the laser. We studied further the influence of the NP size on the NP/PS coupling by investigating the fabrication and fluorescence measurement of Au NPs of different sizes: 10, 30, 50, 80, and 100 nm. The photon collection enhancements in each case were 12.9 +/- 2.5, 12.6 +/- 5.6, 3.9 +/- 2.7, 5.9 +/- 4.4, and 6.6 +/- 5.1 times, respectively. The gain in fluorescence could reach up to 36.6 times for 10-nm gold NPs.
Microultrasound characterisation of ex vivo porcine tissue for ultrasound capsule endoscopy
NASA Astrophysics Data System (ADS)
Lay, H. S.; Cox, B. F.; Sunoqrot, M.; Démoré, C. E. M.; Näthke, I.; Gomez, T.; Cochran, S.
2017-01-01
Gastrointestinal (GI) disease development and progression is often characterised by cellular and tissue architectural changes within the mucosa and sub-mucosa layers. Current clinical capsule endoscopy and other approaches are heavily reliant on optical techniques which cannot detect disease progression below the surface layer of the tissue. To enhance the ability of clinicians to detect cellular changes earlier and more confidently, both quantitative and qualitative microultrasound (μUS) techniques are investigated in healthy ex vivo porcine GI tissue. This work is based on the use of single-element, focussed μUS transducers made with micromoulded piezocomposite operating at around 48 MHz. To explore the possibility that μUS can detect Crohn’s disease and other inflammatory bowel diseases, ex vivo porcine small bowel tissue samples were cannulised and perfused with phosphate-buffered saline followed by various dilutions of polystyrene microspheres. Comparison with fluorescent imaging showed that the microspheres had infiltrated the microvasculature of the samples and that μUS was able to successfully detect this as a mimic of inflammation. Samples without microspheres were analysed using quantitative ultrasound to assess mechanical properties. Attenuation coefficients of 1.78 ± 0.66 dB/mm and 1.92 ± 0.77 dB/mm were obtained from reference samples which were surgically separated from the muscle layer. Six intact samples were segmented using a software algorithm and the acoustic impedance, Z, for varying tissue thicknesses, and backscattering coefficient, BSC, were calculated using the reference attenuation values and tabulated.
Safe biodegradable fluorescent particles
Martin, Sue I [Berkeley, CA; Fergenson, David P [Alamo, CA; Srivastava, Abneesh [Santa Clara, CA; Bogan, Michael J [Dublin, CA; Riot, Vincent J [Oakland, CA; Frank, Matthias [Oakland, CA
2010-08-24
A human-safe fluorescence particle that can be used for fluorescence detection instruments or act as a safe simulant for mimicking the fluorescence properties of microorganisms. The particle comprises a non-biological carrier and natural fluorophores encapsulated in the non-biological carrier. By doping biodegradable-polymer drug delivery microspheres with natural or synthetic fluorophores, the desired fluorescence can be attained or biological organisms can be simulated without the associated risks and logistical difficulties of live microorganisms.
Preparation of "Cauliflower-Like" ZnO Micron-Sized Particles.
Gordon, Tamar; Grinblat, Judith; Margel, Shlomo
2013-11-14
Porous polydivinyl benzene (PDVB) microspheres of narrow size distribution were formed by a single-step swelling process of template uniform polystyrene microspheres with divinyl benzene (DVB), followed by polymerization of the DVB within the swollen template microspheres. The PDVB porous particles were then formed by dissolution of the template polystyrene polymer. Unique "cauliflower-like" ZnO microparticles were prepared by the entrapping of the ZnO precursor ZnCl₂ in the PDVB porous microspheres under vacuum, followed by calcination of the obtained ZnCl₂-PDVB microspheres in an air atmosphere. The morphology, crystallinity and fluorescence properties of those ZnO microparticles were characterized. This "cauliflower-like" shape ZnO particles is in contrast to a previous study demonstrated the preparation of spherical shaped porous ZnO and C-ZnO microparticles by a similar method, using zinc acetate (ZnAc) as a precursor. Two diverted synthesis mechanisms for those two different ZnO microparticles structures are proposed, based on studies of the distribution of each of the ZnO precursors within the PDVB microspheres.
Automatic vision system for analysis of microscopic behavior of flow and transport in porous media
NASA Astrophysics Data System (ADS)
Rashidi, Mehdi; Dehmeshki, Jamshid; Dickenson, Eric; Daemi, M. Farhang
1997-10-01
This paper describes the development of a novel automated and efficient vision system to obtain velocity and concentration measurement within a porous medium. An aqueous fluid lace with a fluorescent dye to microspheres flows through a transparent, refractive-index-matched column packed with transparent crystals. For illumination purposes, a planar sheet of laser passes through the column as a CCD camera records all the laser illuminated planes. Detailed microscopic velocity and concentration fields have been computed within a 3D volume of the column. For measuring velocities, while the aqueous fluid, laced with fluorescent microspheres, flows through the transparent medium, a CCD camera records the motions of the fluorescing particles by a video cassette recorder. The recorded images are acquired automatically frame by frame and transferred to the computer for processing, by using a frame grabber an written relevant algorithms through an RS-232 interface. Since the grabbed image is poor in this stage, some preprocessings are used to enhance particles within images. Finally, these enhanced particles are monitored to calculate velocity vectors in the plane of the beam. For concentration measurements, while the aqueous fluid, laced with a fluorescent organic dye, flows through the transparent medium, a CCD camera sweeps back and forth across the column and records concentration slices on the planes illuminated by the laser beam traveling simultaneously with the camera. Subsequently, these recorded images are transferred to the computer for processing in similar fashion to the velocity measurement. In order to have a fully automatic vision system, several detailed image processing techniques are developed to match exact images that have different intensities values but the same topological characteristics. This results in normalized interstitial chemical concentrations as a function of time within the porous column.
High speed fluorescence imaging with compressed ultrafast photography
NASA Astrophysics Data System (ADS)
Thompson, J. V.; Mason, J. D.; Beier, H. T.; Bixler, J. N.
2017-02-01
Fluorescent lifetime imaging is an optical technique that facilitates imaging molecular interactions and cellular functions. Because the excited lifetime of a fluorophore is sensitive to its local microenvironment,1, 2 measurement of fluorescent lifetimes can be used to accurately detect regional changes in temperature, pH, and ion concentration. However, typical state of the art fluorescent lifetime methods are severely limited when it comes to acquisition time (on the order of seconds to minutes) and video rate imaging. Here we show that compressed ultrafast photography (CUP) can be used in conjunction with fluorescent lifetime imaging to overcome these acquisition rate limitations. Frame rates up to one hundred billion frames per second have been demonstrated with compressed ultrafast photography using a streak camera.3 These rates are achieved by encoding time in the spatial direction with a pseudo-random binary pattern. The time domain information is then reconstructed using a compressed sensing algorithm, resulting in a cube of data (x,y,t) for each readout image. Thus, application of compressed ultrafast photography will allow us to acquire an entire fluorescent lifetime image with a single laser pulse. Using a streak camera with a high-speed CMOS camera, acquisition rates of 100 frames per second can be achieved, which will significantly enhance our ability to quantitatively measure complex biological events with high spatial and temporal resolution. In particular, we will demonstrate the ability of this technique to do single-shot fluorescent lifetime imaging of cells and microspheres.
Bishai, John M; Blood, Arlin B; Hunter, Christian J; Longo, Lawrence D; Power, Gordon G
2003-01-01
This study was undertaken to compare microsphere and laser Doppler flowmetry techniques for the measurement of cerebral blood flow, to assess the effect of probe implantation at the tip of the sensing probe and to measure brain tissue PO2 (tPO2) in response to acute hypoxia. Fetal sheep of ≈131 days gestation (n = 8) were chronically instrumented with bilateral laser Doppler probes in the parietal cortices and catheters for injection of fluorescent microspheres. Five days after surgery fetuses were subjected to 1 h periods of baseline control breathing, hypoxia and recovery. Microspheres were injected 10 min prior to and 10, 30, 50 and 120 min after initiation of hypoxia. Microspheres were counted in four 12 mm3 tissue samples from each hemisphere, the tip of the laser Doppler probe being positioned in the centre of one of the cubes. The cube containing the probe tip was also subdivided into 4 mm3 pieces of tissue. In response to hypoxia, fetal arterial PO2 declined from 21 ± 2 to 12 ± 1 Torr and brain tissue PO2 fell from 10 ± 1 to a nadir of 1 ± 1 Torr. Each method detected a significant increase in CBF that reached a maximum after 30–45 min, although the increase of flow measured by laser Doppler flowmetry was less than that measured by spheres after 10 and 30 min (P < 0.05). Microspheres did not detect altered flow at the probe tip or heterogeneity of flow in surrounding volumes of cortical tissue. In summary, laser Doppler flowmetry is a useful measure of continuous relative changes of CBF in the chronically instrumented fetal sheep. Flow compensations in acute hypoxia are not adequate to sustain O2 delivery, and other compensations, including reduced metabolic rate, are possible. PMID:12563011
Controlling Release Kinetics of PLG Microspheres Using a Manufacturing Technique
NASA Astrophysics Data System (ADS)
Berchane, Nader
2005-11-01
Controlled drug delivery offers numerous advantages compared with conventional free dosage forms, in particular: improved efficacy and patient compliance. Emulsification is a widely used technique to entrap drugs in biodegradable microspheres for controlled drug delivery. The size of the formed microspheres has a significant influence on drug release kinetics. Despite the advantages of controlled drug delivery, previous attempts to achieve predetermined release rates have seen limited success. This study develops a tool to tailor desired release kinetics by combining microsphere batches of specified mean diameter and size distribution. A fluid mechanics based correlation that predicts the average size of Poly(Lactide-co-Glycolide) [PLG] microspheres from the manufacturing technique, is constructed and validated by comparison with experimental results. The microspheres produced are accurately represented by the Rosin-Rammler mathematical distribution function. A mathematical model is formulated that incorporates the microsphere distribution function to predict the release kinetics from mono-dispersed and poly-dispersed populations. Through this mathematical model, different release kinetics can be achieved by combining different sized populations in different ratios. The resulting design tool should prove useful for the pharmaceutical industry to achieve designer release kinetics.
Harvey, Ronald W.; Metge, David W.; Sheets, Rodney A.; Jasperse, Jay
2011-01-01
A major benefit of riverbank filtration (RBF) is that it provides a relatively effective means for pathogen removal. There is a need to conduct more injection-and-recovery transport studies at operating RBF sites in order to properly assess the combined effects of the site heterogeneities and ambient physicochemical conditions, which are difficult to replicate in the lab. For field transport studies involving pathogens, there is considerable interest in using fluorescent carboxylated microspheres (FCM) as surrogates, because they are chemically inert, negatively charged, easy to detect, available in a wide variety of sizes, and have been found to be nonhazardous in tracer applications. Although there have been a number of in-situ studies comparing the subsurface transport behaviors of FCM to those of bacteria and viruses, much less is known about their suitability for investigations of protozoa. Oocysts of the intestinal protozoan pathogen Cryptosporidium spp are of particular concern for many RBF operations because of their ubiquity and persistence in rivers and high resistance to chlorine disinfection. Although microspheres often have proven to be less-than-ideal analogs for capturing the abiotic transport behavior of viruses and bacteria, there is encouraging recent evidence regarding use of FCM as surrogates for C. parvum oocysts. This chapter discusses the potential of fluorescent microspheres as safe and easy-to-detect surrogates for evaluating the efficacy of RBF operations for removing pathogens, particularly Cryptosporidium, from source waters at different points along the flow path.
Digital barcodes of suspension array using laser induced breakdown spectroscopy
He, Qinghua; Liu, Yixi; He, Yonghong; Zhu, Liang; Zhang, Yilong; Shen, Zhiyuan
2016-01-01
We show a coding method of suspension array based on the laser induced breakdown spectroscopy (LIBS), which promotes the barcodes from analog to digital. As the foundation of digital optical barcodes, nanocrystals encoded microspheres are prepared with self-assembly encapsulation method. We confirm that digital multiplexing of LIBS-based coding method becomes feasible since the microsphere can be coded with direct read-out data of wavelengths, and the method can avoid fluorescence signal crosstalk between barcodes and analyte tags, which lead to overall advantages in accuracy and stability to current fluorescent multicolor coding method. This demonstration increases the capability of multiplexed detection and accurate filtrating, expanding more extensive applications of suspension array in life science. PMID:27808270
Zhuang, Si-Hui; Guo, Xin-Xin; Wu, Ying-Song; Chen, Zhen-Hua; Chen, Yao; Ren, Zhi-Qi; Liu, Tian-Cai
2016-01-01
The unique photoproperties of quantum dots are promising for potential application in bioassays. In the present study, quantum dots were applied to a luminescent oxygen channeling assay. The reaction system developed in this study was based on interaction of biotin with streptavidin. Carboxyl-modified polystyrene microspheres doped with quantum dots were biotinylated and used as acceptors. Photosensitizer-doped carboxyl-modified polystyrene microspheres were conjugated with streptavidin and used as donors. The results indicated that the singlet oxygen that was released from the donor beads diffused into the acceptor beads. The acceptor beads were then exited via thioxene, and were subsequently fluoresced. To avoid generating false positives, a high concentration (0.01 mg/mL) of quantum dots is required for application in homogeneous immunoassays. Compared to a conventional luminescent oxygen channeling assay, this quantum dots-based technique requires less time, and would be easier to automate and miniaturize because it requires no washing to remove excess labels.
Becker, M.W.; Reimus, P.W.; Vilks, P.
1999-01-01
Understanding colloid transport in ground water is essential to assessing the migration of colloid-size contaminants, the facilitation of dissolved contaminant transport by colloids, in situ bioremediation, and the health risks of pathogen contamination in drinking water wells. Much has been learned through laboratory and field-scale colloid tracer tests, but progress has been hampered by a lack of consistent tracer testing methodology at different scales and fluid velocities. This paper presents laboratory and field tracer tests in fractured rock that use the same type of colloid tracer over an almost three orders-of-magnitude range in scale and fluid velocity. Fluorescently-dyed carboxylate-modified latex (CML) microspheres (0.19 to 0.98 ??m diameter) were used as tracers in (1) a naturally fractured tuff sample, (2) a large block of naturally fractured granite, (3) a fractured granite field site, and (4) another fractured granite/schist field site. In all cases, the mean transport time of the microspheres was shorter than the solutes, regardless of detection limit. In all but the smallest scale test, only a fraction of the injected microsphere mass was recovered, with the smaller microspheres being recovered to a greater extent than the larger microspheres. Using existing theory, we hypothesize that the observed microsphere early arrival was due to volume exclusion and attenuation was due to aggregation and/or settling during transport. In most tests, microspheres were detected using flow cytometry, which proved to be an excellent method of analysis. CML microspheres appear to be useful tracers for fractured rock in forced gradient and short-term natural gradient tests, but longer residence times may result in small microsphere recoveries.Understanding colloid transport in ground water is essential to assessing the migration of colloid-size contaminants, the facilitation of dissolved contaminant transport by colloids, in situ bioremediation, and the health risks of pathogen contamination in drinking water wells. Much has been learned through laboratory and field-scale colloid tracer tests, but progress has been hampered by a lack of consistent tracer testing methodology at different scales and fluid velocities. This paper presents laboratory and field tracer tests in fractured rock that use the same type of colloid tracer over an almost three orders-of-magnitude range in scale and fluid velocity. Fluorescently-dyed carboxylate-modified latex (CML) microspheres (0.19 to 0.98 ??m diameter) were used as tracers in (1) a naturally fractured tuff sample, (2) a large block of naturally fractured granite, (3) a fractured granite field site, and (4) another fractured granite/schist field site. In all cases, the mean transport time of the microspheres was shorter than the solutes, regardless of detection limit. In all but the smallest scale test, only a fraction of the injected microsphere mass was recovered, with the smaller microspheres being recovered to a greater extent than the larger microspheres. Using existing theory, we hypothesize that the observed microsphere early arrival was due to volume exclusion and attenuation was due to aggregation and/or settling during transport. In most tests, microspheres were detected using flow cytometry, which proved to be an excellent method of analysis. CML microspheres appear to be useful tracers for fractured rock in forced gradient and short-term natural gradient tests, but longer residence times may result in small microsphere recoveries.
Measurement of Scattering and Absorption Cross Sections of Dyed Microspheres
Gaigalas, Adolfas K; Choquette, Steven; Zhang, Yu-Zhong
2013-01-01
Measurements of absorbance and fluorescence emission were carried out on aqueous suspensions of polystyrene (PS) microspheres with a diameter of 2.5 µm using a spectrophotometer with an integrating sphere detector. The apparatus and the principles of measurements were described in our earlier publications. Microspheres with and without green BODIPY@ dye were measured. Placing the suspension inside an integrating sphere (IS) detector of the spectrophotometer yielded (after a correction for fluorescence emission) the absorbance (called A in the text) due to absorption by BODIPY@ dye inside the microsphere. An estimate of the absorbance due to scattering alone was obtained by subtracting the corrected BODIPY@ dye absorbance (A) from the measured absorbance of a suspension placed outside the IS detector (called A1 in the text). The absorption of the BODIPY@ dye inside the microsphere was analyzed using an imaginary index of refraction parameterized with three Gaussian-Lorentz functions. The Kramer-Kronig relation was used to estimate the contribution of the BODIPY@ dye to the real part of the microsphere index of refraction. The complex index of refraction, obtained from the analysis of A, was used to analyze the absorbance due to scattering ((A1- A) in the text). In practice, the analysis of the scattering absorbance, A1-A, and the absorbance, A, was carried out in an iterative manner. It was assumed that A depended primarily on the imaginary part of the microsphere index of refraction with the other parameters playing a secondary role. Therefore A was first analyzed using values of the other parameters obtained from a fit to the absorbance due to scattering, A1-A, with the imaginary part neglected. The imaginary part obtained from the analysis of A was then used to reanalyze A1-A, and obtain better estimates of the other parameters. After a few iterations, consistent estimates were obtained of the scattering and absorption cross sections in the wavelength region 300 nm to 800 nm. PMID:26401422
[Preparation of citrulline microspheres by spray drying technique for colonic targeting].
Bahri, S; Zerrouk, N; Lassoued, M-A; Tsapis, N; Chaumeil, J-C; Sfar, S
2014-03-01
Citrulline is an amino acid that becomes essential in situations of intestinal insufficiency such as short bowel syndrome. It is therefore interesting to provide the patients with dosage forms for routing citrulline to the colon. The aim of this work is to formulate microspheres of citrulline for colonic targeting by the technique of spray drying. Eudragit(®) FS 30D was selected as polymer to encapsulate citrulline using the spray drying technique. Citrulline and Eudragit(®) FS 30D were dissolved in water and ethanol, respectively. The aqueous and the ethanolic solutions were then mixed in 1:2 (v/v) ratio. Microspheres were obtained by nebulizing the citrulline-Eudragit(®) FS 30D solution using a Mini spray dryer equipped with a 0.7mm nozzle. The microspheres have been formulated using citrulline and Eudragit(®) FS 30D. The size distribution of microspheres was determined by light diffraction. The morphology of the microspheres was studied by electron microscopy. Manufacturing yields, encapsulation rate and dissolution profiles were also studied. The microspheres obtained had a spherical shape with a smooth surface and a homogeneous size except for the microspheres containing the highest concentration of polymer (90 %). The formulation showed that the size and morphology of the microspheres are influenced by the polymer concentration. Manufacturing yields were about 51 % but encapsulation rate were always very high (above 90 %). The in vitro dissolution study showed that the use of the Eudragit(®) FS 30D under these conditions is not appropriate to change the dissolution profile of the citrulline. This technique has led to the formulation of microspheres with good physical properties in terms of morphology and size. The compression of the microspheres should help to control citrulline release for colonic targeting. Copyright © 2013 Elsevier Masson SAS. All rights reserved.
NASA Astrophysics Data System (ADS)
Zhang, Jing; Tian, Yu; Ling, Lu-Ting; Yin, Su-Na; Wang, Cai-Feng; Chen, Su
2014-12-01
Versatile hydrogel-based nanocrystal (NC) microreactors were designed in this work for the construction of uniform fluorescence colloidal photonic crystal (CPC) supraballs. The hydrogel-based microspheres with sizes ranging from 150 to 300 nm were prepared by seeded copolymerization of acrylic acid and 2-hydroxyethyl methacrylate with micrometer-sized PS seed particles. As an independent NC microreactor, the as-synthesized hydrogel microsphere can effectively capture the guest cadmium ions due to the abundant carboxyl groups inside. Followed by the introduction of chalcogenides, in situ generation of higher-uptake NCs with sizes less than 5 nm was finally realized. Additionally, with the aid of the microfluidic device, the as-obtained NC-latex hybrids can be further self-assembled to bi-functional CPC supraballs bearing brilliant structural colors and uniform fluorescence. This research offers an alternative way to finely bind CPCs with NCs, which will facilitate progress in fields of self-assembled functional colloids and photonic materials.
FORMULATION AND EVALUATION OF MICROSPHERES CONTAINING LOSARTAN POTASSIUM BY SPRAY-DRYING TECHNIQUE.
Balwierz, Radoslaw; Jankowski, Andrzej; Jasinska, Agata; Marciniak, Dominik; Pluta, Janusz
2016-09-01
Despite numerous applications of microspheres, few works devoted to the preparation of microspheres containing cardiac medications have been published. This study presents the potential of receiving microspheres containing losartan potassium, based on a matrix containing Eudragit L30D55. The study focuses on the possibilities of controlled release of losartan potassium from microspheres in order to reduce the dosage frequency, and also provides information on the effect of the addition of excipients to the quality of the microspheres. Microspheres are monolithic, porous or smooth microparticles ranging from 1 to 500 microns in size. For the preparation of microspheres containing losartan potassium, the spray-drying method was used. The performed study confirmed that the spray-drying technology used to obtain microspheres meets the criteria of size and morphology of the microparticles. The assessment of the kinetics of losartan potassium release from the examined microspheres demonstrated that the release profile followed the first- and/or zero-order kinetics. The use of spray-drying techniques as well as Eudragit L30D55 polymer matrix to obtain the microspheres containing losartan potassium makes it possible to obtain a product with the required particle morphology and particle size ensuring the release of the active substance up to 12 h.
Fiber-optic microsphere-based antibody array for the analysis of inflammatory cytokines in saliva.
Blicharz, Timothy M; Siqueira, Walter L; Helmerhorst, Eva J; Oppenheim, Frank G; Wexler, Philip J; Little, Frédéric F; Walt, David R
2009-03-15
Antibody microarrays have emerged as useful tools for high-throughput protein analysis and candidate biomarker screening. We describe here the development of a multiplexed microsphere-based antibody array capable of simultaneously measuring 10 inflammatory protein mediators. Cytokine-capture microspheres were fabricated by covalently coupling monoclonal antibodies specific for cytokines of interest to fluorescently encoded 3.1 microm polymer microspheres. An optical fiber bundle containing approximately 50,000 individual 3.1 microm diameter fibers was chemically etched to create microwells in which cytokine-capture microspheres could be deposited. Microspheres were randomly distributed in the wells to produce an antibody array for performing a multiplexed sandwich immunoassay. The array responded specifically to recombinant cytokine solutions in a concentration-dependent fashion. The array was also used to examine endogenous mediator patterns in saliva supernatants from patients with pulmonary inflammatory diseases such as asthma and chronic obstructive pulmonary disease (COPD). This array technology may prove useful as a laboratory-based platform for inflammatory disease research and diagnostics, and its small footprint could also enable integration into a microfluidic cassette for use in point-of-care testing.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Hubbard, L; Ziemer, B; Lipinski, J
Purpose: To evaluate the accuracy of a low-dose, first-pass-analysis (FPA) dynamic computed tomography angiography and perfusion (CTAP) technique, for whole-organ anatomical and functional assessment of coronary artery disease (CAD). Methods: An angioplasty balloon was advanced into the left anterior descending (LAD) coronary artery of five swine (35–45 kg) to induce several levels of stenosis at maximal hyperemia (intracoronary adenosine, 240 µg/min). Reference fluorescence microspheres and intravenous contrast (370 mg/mL iodine, 25 mL, 7 mL/s) were injected centrally and dynamic imaging was performed using a 320-slice CT scanner at 100 kVp and 200 mA. Twenty volume scans were acquired per stenosismore » level to capture complete aortic and myocardial enhancement curves, but only two volume scans were used for whole-organ dynamic FPA CTAP measurement. All CTAP measurements in the LAD were compared to the reference microsphere perfusion measurements using linear regression, concordance correlation, and Bland-Altman analysis. Results: The result of dynamic FPA CTAP measurement in the LAD was in good agreement with the reference microsphere perfusion measurement (P-CTAP = 1.01 P-MICRO + 0.16, R{sup 2} = 0.95). The root mean square error (RMSE) and difference (RMSD) of measurement were 0.51 mL/min/g and 0.47 mL/min/g, respectively. Bland-Altman analysis demonstrated negligible systematic measurement bias. Additionally, the concordance correlation coefficient (CCC) was found to be ρ = 0.97, indicating excellent agreement between dynamic FPA CTAP measurement and the reference microsphere perfusion measurement. Lastly, the effective dose of the proposed technique using the “simulated” two-volume scan CTAP acquisition protocol was 2.6 mSv; much lower than the ∼10 mSv effective dose of current dynamic CTP techniques alone. Conclusion: The results indicate the potential for significant improvements in CAD assessment through low-dose, quantitative dynamic FPA CTAP. Such improvements are afforded through whole-organ CT scanning technology, and have the potential to improve patient outcomes and lead to healthier patient lives. Conflict of Interest (only if applicable): Grant funding from Toshiba America Medical Systems.« less
Wang, Ling; Chen, Dong; Gutierrez-Cuevas, Karla G; Bisoyi, Hari Krishna; Fan, Jing; Zola, Rafael S; Li, Guoqiang; Urbas, Augustine M; Bunning, Timothy J; Weitz, David A; Li, Quan
2017-01-01
Optically reconfigurable monodisperse chiral microspheres of self-organized helical superstructures with dynamic chirality were fabricated via a capillary-based microfluidic technique. Light-driven handedness-invertible transformations between different configurations of microspheres were vividly observed and optically tunable RGB photonic cross-communications among the microspheres were demonstrated.
Solvent/Non-Solvent Sintering To Make Microsphere Scaffolds
NASA Technical Reports Server (NTRS)
Laurencin, Cato T.; Brown, Justin L.; Nair, Lakshmi
2011-01-01
A solvent/non-solvent sintering technique has been devised for joining polymeric microspheres to make porous matrices for use as drug-delivery devices or scaffolds that could be seeded with cells for growing tissues. Unlike traditional sintering at elevated temperature and pressure, this technique is practiced at room temperature and pressure and, therefore, does not cause thermal degradation of any drug, protein, or other biochemical with which the microspheres might be loaded to impart properties desired in a specific application. Also, properties of scaffolds made by this technique are more reproducible than are properties of comparable scaffolds made by traditional sintering. The technique involves the use of two miscible organic liquids: one that is and one that is not a solvent for the affected polymer. The polymeric microspheres are placed in a mold having the size and shape of the desired scaffold, then the solvent/non-solvent mixture is poured into the mold to fill the void volume between the microspheres, then the liquid mixture is allowed to evaporate. Some of the properties of the resulting scaffold can be tailored through choice of the proportions of the liquids and the diameter of the microspheres.
Design of polymeric immunomicrospheres for cell labelling and cell separation
NASA Technical Reports Server (NTRS)
Rembaum, A.; Margel, S.
1978-01-01
Synthesis of several classes of hydrophylic microspheres applied to cell labeling and cell separation is described. Five classes of cross-linked microspheres with functional groups such as carboxyl, hydroxyl, amide and/or pyridine groups were synthesized. These functional groups were used to bind covalently antibodies and other proteins to the surface of the microspheres. To optimize the derivatisation technique, polyglutaraldehyde immunomicrospheres were prepared and utilized. Specific populations of human and murine lymphocytes were labelled with microspheres synthesized by the emulsion of the ionizing radiation technique. The labelling of the cells by means of microspheres containing an iron core produced successful separation of B from T lymphocytes by means of a magnetic field.
Mobilization of microspheres from a fractured soil during intermittent infiltration events
Mohanty, Sanjay; Bulicek, Mark; Metge, David W.; Harvey, Ronald W.; Ryan, Joseph N.; Boehm, Alexandria B.
2015-01-01
Pathogens or biocolloids mobilized in the vadose zone may consequently contaminate groundwater. We found that microspheres were mobilized from a fractured soil during intermittent rainfall and the mobilization was greater when the microsphere size was larger and when the soil had greater water permeability.The vadose zone filters pathogenic microbes from infiltrating water and consequently protects the groundwater from possible contamination. In some cases, however, the deposited microbes may be mobilized during rainfall and migrate into the groundwater. We examined the mobilization of microspheres, surrogates for microbes, in an intact core of a fractured soil by intermittent simulated rainfall. Fluorescent polystyrene microspheres of two sizes (0.5 and 1.8 mm) and Br− were first applied to the core to deposit the microspheres, and then the core was subjected to three intermittent infiltration events to mobilize the deposited microspheres. Collecting effluent samples through a 19-port sampler at the base of the core, we found that water flowed through only five ports, and the flow rates varied among the ports by a factor of 12. These results suggest that flow paths leading to the ports had different permeabilities, partly due to macropores. Although 40 to 69% of injected microspheres were retained in the core during their application, 12 to 30% of the retained microspheres were mobilized during three intermittent infiltration events. The extent of microsphere mobilization was greater in flow paths with greater permeability, which indicates that macropores could enhance colloid mobilization during intermittent infiltration events. In all ports, the 1.8-mm microspheres were mobilized to a greater extent than the 0.5-mm microspheres, suggesting that larger colloids are more likely to mobilize. These results are useful in assessing the potential of pathogen mobilization and colloid-facilitated transport of contaminants in the subsurface under natural infiltration events.
Ingestion and transfer of microplastics in the planktonic food web.
Setälä, Outi; Fleming-Lehtinen, Vivi; Lehtiniemi, Maiju
2014-02-01
Experiments were carried out with different Baltic Sea zooplankton taxa to scan their potential to ingest plastics. Mysid shrimps, copepods, cladocerans, rotifers, polychaete larvae and ciliates were exposed to 10 μm fluorescent polystyrene microspheres. These experiments showed ingestion of microspheres in all taxa studied. The highest percentage of individuals with ingested spheres was found in pelagic polychaete larvae, Marenzelleria spp. Experiments with the copepod Eurytemora affinis and the mysid shrimp Neomysis integer showed egestion of microspheres within 12 h. Food web transfer experiments were done by offering zooplankton labelled with ingested microspheres to mysid shrimps. Microscopy observations of mysid intestine showed the presence of zooplankton prey and microspheres after 3 h incubation. This study shows for the first time the potential of plastic microparticle transfer via planktonic organisms from one trophic level (mesozooplankton) to a higher level (macrozooplankton). The impacts of plastic transfer and possible accumulation in the food web need further investigations. Copyright © 2013 Elsevier Ltd. All rights reserved.
Development of wide-angle 2D light scattering static cytometry
NASA Astrophysics Data System (ADS)
Xie, Linyan; Liu, Qiao; Shao, Changshun; Su, Xuantao
2016-10-01
We have recently developed a 2D light scattering static cytometer for cellular analysis in a label-free manner, which measures side scatter (SSC) light in the polar angular range from 79 to 101 degrees. Compared with conventional flow cytometry, our cytometric technique requires no fluorescent labeling of the cells, and static cytometry measurements can be performed without flow control. In this paper we present an improved label-free static cytometer that can obtain 2D light scattering patterns in a wider angular range. By illuminating the static microspheres on chip with a scanning optical fiber, wide-angle 2D light scattering patterns of single standard microspheres with a mean diameter of 3.87 μm are obtained. The 2D patterns of 3.87 μm microspheres contain both large-angle forward scatter (FSC) and SSC light in the polar angular range from 40 to 100 degrees, approximately. Experimental 2D patterns of 3.87 μm microspheres are in good agreement with Mie theory simulated ones. The wide-angle light scattering measurements may provide a better resolution for particle analysis as compared with the SSC measurements. Two dimensional light scattering patterns of HL-60 human acute leukemia cells are obtained by using our static cytometer. Compared with SSC 2D light scattering patterns, wide-angle 2D patterns contain richer information of the HL-60 cells. The obtaining of 2D light scattering patterns in a wide angular range could help to enhance the capabilities of our label-free static cytometry for cell analysis.
Characterization of Buoyant Fluorescent Particles for Field Observations of Water Flows
Tauro, Flavia; Aureli, Matteo; Porfiri, Maurizio; Grimaldi, Salvatore
2010-01-01
In this paper, the feasibility of off-the-shelf buoyant fluorescent microspheres as particle tracers in turbid water flows is investigated. Microspheres’ fluorescence intensity is experimentally measured and detected in placid aqueous suspensions of increasing concentrations of clay to simulate typical conditions occurring in natural drainage networks. Experiments are conducted in a broad range of clay concentrations and particle immersion depths by using photoconductive cells and image-based sensing technologies. Results obtained with both methodologies exhibit comparable trends and show that the considered particles are fairly detectable in critically turbid water flows. Further information on performance and integration of the studied microspheres in low-cost measurement instrumentation for field observations is obtained through experiments conducted in a custom built miniature water channel. This experimental characterization provides a first assessment of the feasibility of commercially available buoyant fluorescent beads in the analysis of high turbidity surface water flows. The proposed technology may serve as a minimally invasive sensing system for hazardous events, such as pollutant diffusion in natural streams and flash flooding due to extreme rainfall. PMID:22163540
DNA polymorphism identity determination using flow cytometry
Nolan, John P.; White, P. Scott; Cai, Hong
2001-01-01
DNA polymorphism identity determination using flow cytometry. Primers designed to be immobilized on microspheres are allowed to anneal to the DNA strand under investigation, and are extended by either DNA polymerase using fluorescent dideoxynucleotides or ligated by DNA ligase to fluorescent reporter oligonucleotides. The fluorescence of either the dideoxynucleotide or the reporter oligonucleotide attached to the immobilized primer is measured by flow cytometry, thereby identifying the nucleotide polymorphism on the DNA strand.
Wu, Fei; Zhang, Kai-Qiang; Bai, Bo; Wang, Hong-Lun; Suo, You-Rui
2015-02-01
In this work, the adsorption potential of TiO2@ yeast composite microspheres to remove Fluorescent Whitening Agent-VBL (FWA-VBL) from aqueous solution was investigated using fixed-bed adsorption column. The effects of pH(2.0-8.0), bed height (1-3 cm), inlet concentration (20-80 mg x L(-1)) and feed flow rate (5-11 mL x min(-1)) on the breakthrough characteristics of the adsorption system were determined. The results showed that the highest bed capacity of 223.80 mg x g(-1) was obtained under the condition of pH 2.0, 80 mg x L(-1) inlet dye concentration, 1.0 cm bed height and 5 mL x min(-1) flow rate. The adsorption data were fitted to three well-established fixed-bed adsorption models, namely, BDST model, Thomas model and Yoon-Nelson model. The results fitted well to the three models with coefficients of correlation R2 > 0.980 in different conditions. The TiO2 @ yeast composite microspheres have desired regeneration ability and could be reused for four times.
Richards, Elaine H; Wontner-Smith, Tim; Bradish, Hannah; Dani, M Paulina; Cotterill, Jane V
2015-09-01
The objective was to develop an environmentally favourable microcapsule suitable for delivery of proteinaceous bioactive agents ('bioinsecticides') to pest insects. Utilising feeding bioassays, we determined that microspheres made of alginate can be produced in a variety of sizes and are palatable and non-toxic to larvae of the lepidopteran pest Lacanobia oleracea. Dehydrated microspheres were also readily ingested by larvae. Using a novel feeding bioassay and alginate microspheres containing a fluorescent marker material (coumarin 7 encapsulated in styrene maleic anhydride beads), we determined that the microspheres successfully deliver the marker to the insect gut. Moreover, the alginate microspheres rapidly break down in the alkaline conditions of the insect gut and release their contents, the beads passing through the gut in 2-3 h. Using bovine serum albumin as a test protein and western blotting, it was determined that alginate can successfully encapsulate protein, and that the microspheres can be stored in a CaCl2 solution for up to 24 days without extensive leakage. Importantly, it was also determined that alginate and the microsphere-making procedure developed do not inactivate rVPr1 (an insect immunosuppressive protein and potential bioinsecticide). An alginate-based microsphere has potential to deliver the proteinaceous bioactive rVPr1 to pest insects. © 2014 Crown copyright. Pest Management Science © 2014 Society of Chemical Industry.
2016-01-01
A combined surface treatment (i.e., surface grafting and a layer-by-layer (LbL) approach) is presented to create advanced biomaterials, i.e., 3D poly(l-lactide) (PLLA) microsphere scaffolds, at room temperature. The grafted surface plays a crucial role in assembling polyelectrolyte multilayers (PEMs) onto the surface of the microspheres, thus improving the physicochemical properties of the 3D microsphere scaffolds. The grafted surface of the PLLA microspheres demonstrates much better PEM adsorption, improved surface coverage at low pH, and smoother surfaces at high pH compared with those of nongrafted surfaces of PLLA microspheres during the assembly of PEMs. They induce more swelling than nongrafted surfaces after the assembly of the PEMs and exhibit blue emission after functionalization of the microsphere surface with a fluorescent dye molecule. The 3D scaffolds functionalized with and without nanosheets not only exhibit good mechanical performance similar to the compressive modulus of cancellous bone but also exhibit the porosity required for cancellous bone regeneration. The magnetic nanoparticle-functionalized 3D scaffolds result in an electrical conductivity in the high range of semiconducting materials (i.e., 1–250 S cm–1). Thus, these 3D microsphere scaffolds fabricated by surface grafting and the LbL approach are promising candidates for bone tissue engineering. PMID:29503506
Magnetic focusing immunosensor for the detection of Salmonella typhimurium in foods
NASA Astrophysics Data System (ADS)
Pivarnik, Philip E.; Cao, He; Letcher, Stephen V.; Pierson, Arthur H.; Rand, Arthur G.
1999-01-01
From 1988 through 1992 Salmonellosis accounted for 27% of the total reported foodborne disease outbreaks and 57% of the outbreaks in which the pathogen was identified. The prevalence of Salmonellosis and the new requirements to monitor the organism as a marker in pathogen reduction programs will drive the need for rapid, on-site testing. A compact fiber optic fluorometer using a red diode laser as an excitation source and fiber probes for analyte detection has been constructed and used to measure Salmonella. The organisms were isolated with anti-Salmonella magnetic beads and were labeled with a secondary antibody conjugated to a red fluorescent dye. The response of the system was proportional to the concentration of Salmonella typhimurium from 3.2 X 105 colony forming units (CFU)/ml to 1.6 X 107 CFU/ml. The system was developed to utilize a fiber-optic magnetic focusing problem that attracted the magnetic microspheres to the surface of a sample chamber directly in front of the excitation and emission fibers. The signal obtained from a homogenous suspension of fluorescent magnetic microspheres was 9 to 10 picowatts. After focusing, the signal from the fluorescent labeled magnetic microspheres increased to 200 picowatts, approximately 20 times greater than the homogeneous suspension. The magnetic focusing assay detected 1.59 X 105 colony forming units/ml of Salmonella typhimurium cultured in growth media. The process of magnetic focusing in front of the fibers has the potential to reduce the background fluorescence from unbound secondary antibodies, eliminating several rinsing steps, resulting in a simple rapid assay.
Nitride Fuel Development Using Cryo-process Technique
DOE Office of Scientific and Technical Information (OSTI.GOV)
O'Brien, Brandi M; Windes, William E
A new cryo-process technique has been developed for the fabrication of advanced fuel for nuclear systems. The process uses a new cryo-processing technique whereby small, porous microspheres (<2000 µm) are formed from sub-micron oxide powder. A simple aqueous particle slurry of oxide powder is pumped through a microsphere generator consisting of a vibrating needle with controlled amplitude and frequency. As the water-based droplets are formed and pass through the microsphere generator they are frozen in a bath of liquid nitrogen and promptly vacuum freeze-dried to remove the water. The resulting porous microspheres consist of half micron sized oxide particles heldmore » together by electrostatic forces and mechanical interlocking of the particles. Oxide powder microspheres ranging from 750 µm to 2000 µm are then converted into a nitride form using a high temperature fluidized particle bed. Carbon black can be added to the oxide powder before microsphere formation to augment the carbothermic reaction during conversion to a nitride. Also, the addition of ethyl alcohol to the aqueous slurry reduces the surface tension energy of the droplets resulting in even smaller droplets forming in the microsphere generator. Initial results from this new process indicate a lower impurity contamination in the final nitrides due to the single feed stream of particles, material handling and conversion are greatly simplified, a minimum of waste and personnel exposure are anticipated, and finally the conversion kinetics may be greatly increased because of the small oxide powder size (sub-micron) forming the porous microsphere. Thus far the fabrication process has been successful in demonstrating all of these improvements with surrogate ZrO2 powder. Further tests will be conducted in the future using the technique on UO2 powders.« less
Simultaneous imaging of blood flow dynamics and vascular remodelling during development.
Ghaffari, Siavash; Leask, Richard L; Jones, Elizabeth A V
2015-12-01
Normal vascular development requires blood flow. Time-lapse imaging techniques have revolutionised our understanding of developmental biology, but measuring changes in blood flow dynamics has met with limited success. Ultrasound biomicroscopy and optical coherence tomography can concurrently image vascular structure and blood flow velocity, but these techniques lack the resolution to accurately calculate fluid forces such as shear stress. This is important because hemodynamic forces are biologically active and induce changes in the expression of genes important for vascular development. Regional variations in shear stress, rather than the overall level, control processes such as vessel enlargement and regression during vascular remodelling. We present a technique to concurrently visualise vascular remodelling and blood flow dynamics. We use an avian embryonic model and inject an endothelial-specific dye and fluorescent microspheres. The motion of the microspheres is captured with a high-speed camera and the velocity of the blood flow in and out of the region of interest is quantified by micro-particle image velocitymetry (µPIV). The vessel geometry and flow are used to numerically solve the flow physics with computational fluid dynamics (CFD). Using this technique, we can analyse changes in shear stress, pressure drops and blood flow velocities over a period of 10 to 16 h. We apply this to study the relationship between shear stress and chronic changes in vessel diameter during embryonic development, both in normal development and after TGFβ stimulation. This technique allows us to study the interaction of biomolecular and biomechanical signals during vascular remodelling using an in vivo developmental model. © 2015. Published by The Company of Biologists Ltd.
Microsphere-Based Immunoassay for the Detection of Azaspiracids
Rodríguez, Laura P.; Vilariño, Natalia; Louzao, M. Carmen; Dickerson, Tobin J.; Nicolaou, K. C.; Frederick, Michael O.; Botana, Luis M.
2014-01-01
Azaspiracids (AZAs) are a group of lipophilic toxins discovered in mussels from Ireland in 1995 following a human poisoning incident. Nowadays the regulatory limit for AZAs in many countries is set at 160 Fg of azaspiracid equivalents per kg of shellfish meat. In this work a microsphere-based immunoassay has been developed for the detection of AZAs using a Luminex system. This method is based on the competition between AZA-2 immobilized onto the surface of microspheres and free AZAs for the interaction with a monoclonal anti-azaspiracid antibody (mAb 8F4). In this inhibition immunoassay the amount of mAb 8F4 bound to AZA-2-microspheres was quantified using a phycoerythrin-labeled anti-mouse antibody, and the fluorescence was measured with a Luminex analyzer. Simple acetate/methanol or methanol extractions yielded final extracts with no matrix interferences and adequate recovery rates of 86.5% and 75.8%, respectively. In summary, this work presents, a sensitive and easily performed screening method capable of detecting AZAs at concentrations below the range of the European regulatory limit using a microsphere/flow cytometry system. PMID:24215909
Performance of computer vision in vivo flow cytometry with low fluorescence contrast
NASA Astrophysics Data System (ADS)
Markovic, Stacey; Li, Siyuan; Niedre, Mark
2015-03-01
Detection and enumeration of circulating cells in the bloodstream of small animals are important in many areas of preclinical biomedical research, including cancer metastasis, immunology, and reproductive medicine. Optical in vivo flow cytometry (IVFC) represents a class of technologies that allow noninvasive and continuous enumeration of circulating cells without drawing blood samples. We recently developed a technique termed computer vision in vivo flow cytometry (CV-IVFC) that uses a high-sensitivity fluorescence camera and an automated computer vision algorithm to interrogate relatively large circulating blood volumes in the ear of a mouse. We detected circulating cells at concentrations as low as 20 cells/mL. In the present work, we characterized the performance of CV-IVFC with low-contrast imaging conditions with (1) weak cell fluorescent labeling using cell-simulating fluorescent microspheres with varying brightness and (2) high background tissue autofluorescence by varying autofluorescence properties of optical phantoms. Our analysis indicates that CV-IVFC can robustly track and enumerate circulating cells with at least 50% sensitivity even in conditions with two orders of magnitude degraded contrast than our previous in vivo work. These results support the significant potential utility of CV-IVFC in a wide range of in vivo biological models.
Lee, Dennis; Barnes, Stephen
2010-01-01
The need for new pharmacological agents is unending. Yet the drug discovery process has changed substantially over the past decade and continues to evolve in response to new technologies. There is presently a high demand to reduce discovery time by improving specific lab disciplines and developing new technology platforms in the area of cell-based assay screening. Here we present the developmental concept and early stage testing of the Ab-Sniffer, a novel fiber optic fluorescence device for high-throughput cytotoxicity screening using an immobilized whole cell approach. The fused silica fibers are chemically functionalized with biotin to provide interaction with fluorescently labeled, streptavidin functionalized alginate-chitosan microspheres. The microspheres are also functionalized with Concanavalin A to facilitate binding to living cells. By using lymphoma cells and rituximab in an adaptation of a well-known cytotoxicity protocol we demonstrate the utility of the Ab-Sniffer for functional screening of potential drug compounds rather than indirect, non-functional screening via binding assay. The platform can be extended to any assay capable of being tied to a fluorescence response including multiple target cells in each well of a multi-well plate for high-throughput screening.
Yanagawa, Katsunori; Nunoura, Takuro; McAllister, Sean M.; Hirai, Miho; Breuker, Anja; Brandt, Leah; House, Christopher H.; Moyer, Craig L.; Birrien, Jean-Louis; Aoike, Kan; Sunamura, Michinari; Urabe, Tetsuro; Mottl, Michael J.; Takai, Ken
2013-01-01
During the Integrated Ocean Drilling Program (IODP) Expedition 331 at the Iheya North hydrothermal system in the Mid-Okinawa Trough by the D/V Chikyu, we conducted microbiological contamination tests of the drilling and coring operations. The contamination from the drilling mud fluids was assessed using both perfluorocarbon tracers (PFT) and fluorescent microsphere beads. PFT infiltration was detected from the periphery of almost all whole round cores (WRCs). By contrast, fluorescent microspheres were not detected in hydrothermally active core samples, possibly due to thermal decomposition of the microspheres under high-temperature conditions. Microbial contamination from drilling mud fluids to the core interior subsamples was further characterized by molecular-based evaluation. The microbial 16S rRNA gene phylotype compositions in the drilling mud fluids were mainly composed of sequences of Beta- and Gammaproteobacteria, and Bacteroidetes and not archaeal sequences. The phylotypes that displayed more than 97% similarity to the sequences obtained from the drilling mud fluids were defined as possible contaminants in this study and were detected as minor components of the bacterial phylotype compositions in 13 of 37 core samples. The degree of microbiological contamination was consistent with that determined by the PFT and/or microsphere assessments. This study suggests a constructive approach for evaluation and eliminating microbial contamination during riser-less drilling and coring operations by the D/V Chikyu. PMID:24265628
Zhang, Li; Liang, Shuang; Liu, Ruiqing; Yuan, Tianmeng; Zhang, Shulai; Xu, Zushun; Xu, Haibo
2016-08-01
Molecular imaging is of significant importance for early detection and diagnosis of cancer. Herein, a novel core-shell magnetic microsphere for dual modal magnetic resonance imaging (MRI) and optical imaging was produced by one-pot emulsifier-free emulsion polymerization, which could provide high resolution rate of histologic structure information and realize high sensitive detection at the same time. The synthesized magnetic microspheres composed of cores containing oleic acid (OA) and sodium undecylenate (NaUA) modified Fe3O4 nanoparticles and styrene (St), Glycidyl methacrylate (GMA), and polymerizable lanthanide complexes (Gd(AA)3Phen and Eu(AA)3Phen) polymerized on the surface for outer shells. Fluorescence spectra show characteristic emission peaks from Eu(3+) at 590nm and 615nm and vivid red fluorescence luminescence can be observed by 2-photon confocal scanning laser microscopy (CLSM). In vitro cytotoxicity tests based on the MTT assay demonstrate good cytocompatibility, the composites have longitudinal relaxivity value (r1) of 8.39mM(-1)s(-1) and also have transverse relaxivity value (r2) of 71.18mM(-1)s(-1) at clinical 3.0 T MR scanner. In vitro and in vivo MRI studies exhibit high signal enhancement on both T1- and T2-weighted MR images. These fascinating multifunctional properties suggest that the polymer microspheres have large clinical potential as multi-modal MRI/optical probes. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.
Yanagawa, Katsunori; Nunoura, Takuro; McAllister, Sean M; Hirai, Miho; Breuker, Anja; Brandt, Leah; House, Christopher H; Moyer, Craig L; Birrien, Jean-Louis; Aoike, Kan; Sunamura, Michinari; Urabe, Tetsuro; Mottl, Michael J; Takai, Ken
2013-01-01
During the Integrated Ocean Drilling Program (IODP) Expedition 331 at the Iheya North hydrothermal system in the Mid-Okinawa Trough by the D/V Chikyu, we conducted microbiological contamination tests of the drilling and coring operations. The contamination from the drilling mud fluids was assessed using both perfluorocarbon tracers (PFT) and fluorescent microsphere beads. PFT infiltration was detected from the periphery of almost all whole round cores (WRCs). By contrast, fluorescent microspheres were not detected in hydrothermally active core samples, possibly due to thermal decomposition of the microspheres under high-temperature conditions. Microbial contamination from drilling mud fluids to the core interior subsamples was further characterized by molecular-based evaluation. The microbial 16S rRNA gene phylotype compositions in the drilling mud fluids were mainly composed of sequences of Beta- and Gammaproteobacteria, and Bacteroidetes and not archaeal sequences. The phylotypes that displayed more than 97% similarity to the sequences obtained from the drilling mud fluids were defined as possible contaminants in this study and were detected as minor components of the bacterial phylotype compositions in 13 of 37 core samples. The degree of microbiological contamination was consistent with that determined by the PFT and/or microsphere assessments. This study suggests a constructive approach for evaluation and eliminating microbial contamination during riser-less drilling and coring operations by the D/V Chikyu.
Multitracing Experiment With Solved and Particulate Tracers In An Unsaturated Field Soil
NASA Astrophysics Data System (ADS)
Burkhardt, M.; Kasteel, R.; Vereecken, H.
Solute movement and colloid migration follow preferential flow paths in structured soils at the field scale. The use of microsphreres is a possible option to mimic colloid transport through the vadose zone into the groundwater. We present results of multi- tracing experiments conducted in an Orthic Luvisol using bromide (Br-), the reactive dye tracer Brilliant Blue (BB) and microspheres. The fluorescent microspheres (1 and 10 µm in diameter) were functionalized with a negative surface charge. Eight field plots (about 2 m2) were irrigated with 10 mm and 40 mm during 6 h. Four field plots were sampled directly after the irrgation, the others were exposed for 90 days to natural wheather conditions. Photographs of horizontal cross-sections and disturbed soil sam- ples were taken every 5 to 10 cm down to a depth of 160 cm. Image analysis was used to derive concentration distributions of BB using a calibration relationship between concentration and color spectra. The microspheres were quantified after desorption of the soil samples by fluorescent microscopy and image analysis. We used moment analysis to characterize transport phenomena. We found that transport through the soil matrix was affected by sorption, but all of the applied compounds were transported through preferential flow paths (earthworm burrows) down to a depth of 160 cm irre- spective of their chemical properties. Furthermore, this study shows that microspheres can be used to mimic colloid facilitated transport under unsaturated conditions in a field soil.
Recent advances in testing of microsphere drug delivery systems.
Andhariya, Janki V; Burgess, Diane J
2016-01-01
This review discusses advances in the field of microsphere testing. In vitro release-testing methods such as sample and separate, dialysis membrane sacs and USP apparatus IV have been used for microspheres. Based on comparisons of these methods, USP apparatus IV is currently the method of choice. Accelerated in vitro release tests have been developed to shorten the testing time for quality control purposes. In vitro-in vivo correlations using real-time and accelerated release data have been developed, to minimize the need to conduct in vivo performance evaluation. Storage stability studies have been conducted to investigate the influence of various environmental factors on microsphere quality throughout the product shelf life. New tests such as the floating test and the in vitro wash-off test have been developed along with advancement in characterization techniques for other physico-chemical parameters such as particle size, drug content, and thermal properties. Although significant developments have been made in microsphere release testing, there is still a lack of guidance in this area. Microsphere storage stability studies should be extended to include microspheres containing large molecules. An agreement needs to be reached on the use of particle sizing techniques to avoid inconsistent data. An approach needs to be developed to determine total moisture content of microspheres.
Acrolein Microspheres Are Bonded To Large-Area Substrates
NASA Technical Reports Server (NTRS)
Rembaum, Alan; Yen, Richard C. K.
1988-01-01
Reactive cross-linked microspheres produced under influence of ionizing radiation in aqueous solutions of unsaturated aldehydes, such as acrolein, with sodium dodecyl sulfate. Diameters of spheres depend on concentrations of ingredients. If polystyrene, polymethylmethacrylate, or polypropylene object immersed in solution during irradiation, microspheres become attached to surface. Resulting modified surface has grainy coating with reactivity similar to free microspheres. Aldehyde-substituted-functional microspheres react under mild conditions with number of organic reagents and with most proteins. Microsphere-coated macrospheres or films used to immobilize high concentrations of proteins, enzymes, hormones, viruses, cells, and large number of organic compounds. Applications include separation techniques, clinical diagnostic tests, catalytic processes, and battery separators.
A novel microsphere with a three-layer structure for duodenum-specific drug delivery.
Zhu, Xi; Zhou, Dan; Jin, Yun; Song, Yu-pin; Zhang, Zhi-rong; Huang, Yuan
2011-07-15
Owing to the quick elimination of drug from duodenum and the depth of Helicobacter pylori (H. pylori) colonized in mucus, antibiotic therapy often fails in the eradication of H. pylori infection for duodenal ulcer. A novel duodenum-specific microsphere (DSM) consisting of three-layer structure was developed to enhance the drug concentration and retention time in duodenal mucus layer. Firstly a core-shell mucoadhesive microsphere was prepared with a novel emulsification/coagulation coating method by introducing drug loaded Eudragit cores into a thiolated chitosan mucoadhesive layer. Then the obtained core-shell mucoadhesive microspheres were further coated with hydroxypropyl methylcellulose acetate maleate as the pH-sensitive layer for the trigger of mucoadhesion and drug release in duodenum. From the fluorescence microscopic and scanning electron microscopic images, the three-layer structure was successfully established. The microspheres exhibited a duodenum-specific trigger performance, good mucoadhesive property and pH-dependent drug release. In vivo study performed in rats demonstrated that DSM exhibited about 3-fold augmentation of AUC and about 5-fold augmentation of C(max) for duodenal mucus drug concentration compared with free drug suspension. These results suggest that the three-layer structure microspheres may provide a promising approach for duodenum-targeting drug delivery system. Copyright © 2011 Elsevier B.V. All rights reserved.
Detection of inflammatory cytokines using a fiber optic microsphere immunoassay array
NASA Astrophysics Data System (ADS)
Blicharz, Timothy M.; Walt, David R.
2006-10-01
A multiplexed fiber optic microsphere-based immunoassay array capable of simultaneously measuring five inflammatory cytokines has been developed. Five groups of amine-functionalized 3.1 micron microspheres were internally encoded with five distinct concentrations of a europium dye and converted to cytokine probes by covalently coupling monoclonal capture antibodies specific for human VEGF, IFN-gamma, RANTES, IP-10, and Eotaxin-3 to the microspheres via glutaraldehyde chemistry. The microspheres were pooled and loaded into a 1 mm diameter fiber optic bundle containing ~50,000 individual etched microwells, producing the multiplexed cytokine immunoassay array. Multiple arrays can be created from a single microsphere pool for high throughput sample analysis. Sandwich fluoroimmunoassays were performed by incubating the probe array in a sample, followed by incubation in a mixture of biotin-labeled detection antibodies that are complementary to the five cytokines. Finally, universal detection of each protein was performed using a fluorescence imaging system after briefly immersing the array in a solution of fluorophore-labeled streptavidin. The multiplexed cytokine array has been shown to respond selectively to VEGF, IFNgamma, RANTES, IP-10, and Eotaxin-3, permitting multiplexed quantitative analysis. Ultimately, the multiplexed cytokine array will be utilized to evaluate the potential of using saliva as a noninvasive diagnostic fluid for pulmonary inflammatory diseases such as asthma.
NASA Astrophysics Data System (ADS)
Pai, Rajesh V.; Mollick, P. K.; Kumar, Ashok; Banerjee, J.; Radhakrishna, J.; Chakravartty, J. K.
2016-05-01
UO2 microspheres prepared by internal gelation technique were coated with pyrolytic carbon and silicon carbide using CVD technique. The particles which were not meeting the specifications were rejected. The rejected/failed UO2 based coated particles prepared by CVD technique was used for oxidation and recovery and recycling. The oxidation behaviour of sintered UO2 microspheres coated with different layers of carbon and SiC was studied by thermal techniques to develop a method for recycling and recovery of uranium from the failed/rejected coated particles. It was observed that the complete removal of outer carbon from the spheres is difficult. The crushing of microspheres enabled easier accessibility of oxygen and oxidation of carbon and uranium at 800-1000 °C. With the optimized process of multiple crushing using die & plunger and sieving the broken coated layers, we could recycle around fifty percent of the UO2 microspheres which could be directly recoated. The rest of the particles were recycled using a wet recycling method.
Lee, S; Kim, M S; Kim, J S; Park, H J; Woo, J S; Lee, B C; Hwang, S J
2006-11-01
The purpose of this study was to prepare microspheres loaded with hydrophilic drug, bupivacaine HCl using poly(D,L-lactic-co-glycolic acid) (PLGA) and poly(L-lactic acid) (PLLA). Microspheres were prepared with varying the PLGA/PLLA ratio with two different levels of bupivacaine HCl (5 and 10%) using a supercritical anti-solvent (SAS) technique. Microspheres ranging from 4-10 microm in geometric mean diameter could be prepared, with high loading efficiency. Powder X-ray diffraction (PXRD) revealed that bupivacaine HCl retained its crystalline state within the polymer and was present as a dispersion within the polymer phase after SAS processing. The release of bupivacaine HCl from biodegradable polymer microspheres was rapid up to 4 h, thereafter bupivacaine HCl was continuously and slowly released for at least 7 days according to the PLGA/PLLA ratio and the molecular weight of PLLA.
Sensing and enumerating rare circulating cells with diffuse light
NASA Astrophysics Data System (ADS)
Zettergren, Eric; Vickers, Dwayne; Niedre, Mark
2011-02-01
Detection and quantification of circulating cells in live animals is a challenging and important problem in many areas of biomedical research. Current methods involve extraction of blood samples and counting of cells ex-vivo. Since only small blood volumes are analyzed at specific time points, monitoring of changes in cell populations over time is difficult and rare cells often escape detection. The goal of this research is to develop a method for enumerating very rare circulating cells in the bloodstream non-invasively. This would have many applications in biomedical research, including monitoring of cancer metastasis and tracking of hematopoietic stem cells. In this work we describe the optical configuration of our instrument which allows fluorescence detection of single cells in diffusive media at the mesoscopic scale. Our instrument design consists of two continuous wave laser diode sources and an 8-channel fiber coupled multi-anode photon counting PMT. Fluorescence detector fibers were arranged circularly around the target in a miniaturized ring configuration. Cell-simulating fluorescent microspheres and fluorescently-labeled cells were passed through a limb mimicking phantom with similar optical properties and background fluorescence as a limb of a mouse. Our data shows that we are able to successfully detect and count these with high quantitative accuracy. Future work includes characterization of our instrument using fluorescently labeled cells in-vivo. If successful, this technique would allow several orders of magnitude in vivo detection sensitivity improvement versus current approaches.
Preparation and evaluation of sustained release loxoprofen loaded microspheres.
Venkatesan, P; Manavalan, R; Valliappan, K
2011-06-01
The aim of present study was to formulate and evaluate the loxoprofen loaded Sustained release microspheres by emulsion solvent evaporation technique. Ethylcellulose, a biocompatible polymer is used as the retardant material. The effects of process conditions such as drug loading, polymer type and solvent type on the characteristics of microspheres were investigated. The prepared microspheres were characterized for their particle size and drug loading and drug release. The in-vitro release studies were carried out in phosphate buffer at pH 7.4. The prepared microspheres were white, free flowing and spherical in shape. The drug-loaded microspheres showed 71.2% of entrapment and the in-vitro release studies showed that Loxoprofen microspheres of 1:3 ratios showed better sustained effect over a period of 8 hours.
Preparation and evaluation of sustained release loxoprofen loaded microspheres
Venkatesan, P.; Manavalan, R.; Valliappan, K.
2011-01-01
The aim of present study was to formulate and evaluate the loxoprofen loaded Sustained release microspheres by emulsion solvent evaporation technique. Ethylcellulose, a biocompatible polymer is used as the retardant material. The effects of process conditions such as drug loading, polymer type and solvent type on the characteristics of microspheres were investigated. The prepared microspheres were characterized for their particle size and drug loading and drug release. The in-vitro release studies were carried out in phosphate buffer at pH 7.4. The prepared microspheres were white, free flowing and spherical in shape. The drug-loaded microspheres showed 71.2% of entrapment and the in-vitro release studies showed that Loxoprofen microspheres of 1:3 ratios showed better sustained effect over a period of 8 hours PMID:24826017
Magnetically guiding and orienting integrated chemical sensors
NASA Astrophysics Data System (ADS)
Anker, Jeffrey N.; Lee, Yong-Eun Koo; Kopelman, Raoul
2014-08-01
Fluorescent microsensors for detecting pH and oxygen were positioned and oriented using magnetic tweezers. These multifunctional integrated microsensors were fabricated by physically linking together nano-components including magnetic nanoparticles, fluorescent nanoparticles, and metal hemisphere-shells. Two such microsensors are magnetic roll-shaped polystyrene particles with 120 nm fluorescent oxygen-sensing ormosil nanospheres that are physically pressed ("breaded") into the roll surface, and 4-5 µm fluorescent microspheres that are capped with a 50 nm thick metal hemispherical shell. The magnetic tweezers consisted of an iron wire that was magnetized in an external magnetic field. Rotating this external field oriented and rotated the microsensors.
Peng, Di; Liu, Xiang; Huang, Mengjun; Wang, Dan; Liu, Renlong
2018-04-24
Solid powder fluorescence shows great potential for application in medicine, biology, and engineering, especially in the identification of latent fingermarks in forensic science. However, conventional developing methods suffer from some drawbacks, such as low contrast, low sensitivity, low selectivity, and high toxicity. To conquer these challenges, novel SiO2@C-dot microspheres were prepared via a facile one-pot hydrothermal method by using citric acid as a carbon source and aminosilane as a nitrogen source. Interestingly, the results showed that the resultant powders possess good monodispersity, high fluorescence emission, and resistance to self-quenching. Additionally, the mechanism for the solid-state fluorescence of SiO2@C-dot compounds was also investigated. More importantly, the fingermarks on various surfaces, including transparent glasses, ceramic tiles, transparent plastics, aluminum alloys, plastic cards, painted woods, artificial leathers, and Chinese paper money, developed by the powders have indicated well-defined papillary ridges under a 365 nm UV lamp. The novel strategy of using monodisperse SiO2@C-dot microspheres as a fluorescent label for developing latent fingermarks showed greater advantages compared to conventional methods, which was also demonstrated using the automatic fingerprint identification system. It is simple, rapid, low-cost, nontoxic, and effective, and is expected to be a promising alternative for the development of latent fingerprints in forensic science.
Fantozzi, Anna; Ermolli, Monica; Marini, Massimiliano; Scotti, Domenico; Balla, Branko; Querci, Maddalena; Langrell, Stephen R H; Van den Eede, Guy
2007-02-21
An innovative covalent microsphere immunoassay, based on the usage of fluorescent beads coupled to a specific antibody, was developed for the quantification of the endotoxin Cry1Ab present in MON810 and Bt11 genetically modified (GM) maize lines. In particular, a specific protocol was developed to assess the presence of Cry1Ab in a very broad range of GM maize concentrations, from 0.1 to 100% [weight of genetically modified organism (GMO)/weight]. Test linearity was achieved in the range of values from 0.1 to 3%, whereas fluorescence signal increased following a nonlinear model, reaching a plateau at 25%. The limits of detection and quantification were equal to 0.018 and 0.054%, respectively. The present study describes the first application of quantitative high-throughput immunoassays in GMO analysis.
Photopolymerization Of Levitated Droplets
NASA Technical Reports Server (NTRS)
Rembaum, Alan; Rhim, Won-Kyu; Hyson, Michael T.; Chang, Manchium
1989-01-01
Experimental containerless process combines two established techniques to make variety of polymeric microspheres. In single step, electrostatically-levitated monomer droplets polymerized by ultraviolet light. Faster than multiple-step emulsion polymerization process used to make microspheres. Droplets suspended in cylindrical quadrupole electrostatic levitator. Alternating electrostatic field produces dynamic potential along axis. Process enables tailoring of microspheres for medical, scientific, and industrial applications.
Sivakumar, M; Rao, K Panduranga
2003-05-01
In this work, composite microspheres were prepared from bioactive ceramics such as coralline hydroxyapatite [Ca(10)(PO(4))(6)(OH)(2)] granules, a biodegradable polymer, sodium alginate, and an antibiotic, gentamicin. Previously, we have shown a gentamicin release from coralline hydroxyapatite granules-chitosan composite microspheres. In the present investigation, we attempted to prepare composite microspheres containing coralline hydroxyapatite granules and sodium alginate by the dispersion polymerization technique with gentamicin incorporated by absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. Fourier transform infrared spectra clearly indicated the presence of per-acid of sodium alginate, phosphate, and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs showed that the composite microspheres were spherical in shape and porous in nature. The particle size of composite microspheres was analyzed, and the average size was found to be 15 microns. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns. Copyright 2003 Wiley Periodicals, Inc.
Sivakumar, M; Panduranga Rao, K
2002-08-01
Composite microspheres have been prepared from bioactive ceramics such as coralline hydroxyapatite [CHA, Ca10(PO4)6(OH)2] granules, a biodegradable polymer, gelatin and an antibiotic, gentamicin. In our earlier work, we have shown a gentamicin release from CHA granules--chitosan composite microspheres. In the present investigation, an attempt was made to prepare the composite microspheres containing coralline hydroxyapatite and gelatin (CHA-G), which were prepared by the dispersion polymerization technique and the gentamicin was incorporated by the absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. The Fourier transformed infrared spectrum clearly indicated the presence of amide and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs show that the composite microspheres are spherical in shape and porous in nature. The particle size of composite microspheres was analyzed and the average size was found to be 16 microm. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns.
Zhao, Qian; Piao, Jiafang; Peng, Weipan; Wang, Yang; Zhang, Bo; Gong, Xiaoqun; Chang, Jin
2018-01-31
Identifying the microRNA (miRNA) expression level can provide critical information for early diagnosis of cancers or monitoring the cancer therapeutic efficacy. This paper focused on a kind of gold-nanoparticle-coated polystyrene microbeads (PS@Au microspheres)-based DNA probe as miRNA capture and duplex-specific nuclease (DSN) signal amplification platform based on an RGB value readout for detection of miRNAs. In virtue of the outstanding selectivity and simple experimental operation, 5'-fluorochrome-labeled molecular beacons (MBs) were immobilized on PS@Au microspheres via their 3'-thiol, in the wake of the fluorescence quenching by nanoparticle surface energy transfer (NSET). Target miRNAs were captured by the PS@Au microspheres-based DNA probe through DNA/RNA hybridization. DSN enzyme subsequently selectively cleaved the DNA to recycle the target miRNA and release of fluorophores, thereby triggering the signal amplification with more free fluorophores. The RGB value measurement enabled a detection limit of 50 fM, almost 4 orders of magnitude lower than PS@Au microspheres-based DNA probe detection without DSN. Meanwhile, by different encoding of dyes, miRNA-21 and miRNA-10b were simultaneously detected in the same sample. Considering the ability for quantitation, high sensitivity, and convenient merits, the PS@Au microspheres-based DNA probe and DSN signal amplification platform supplied valuable information for early diagnosis of cancers.
Active Q switching of a fiber laser with a microsphere resonator
NASA Astrophysics Data System (ADS)
Kieu, Khanh; Mansuripur, Masud
2006-12-01
We propose and demonstrate an active Q-switched fiber laser using a high-Q microsphere resonator as the Q-switching element. The laser cavity consists of an Er-doped fiber as the gain medium, a glass microsphere reflector (coupled through a fiber taper) at one end of the cavity, and a fiber Bragg grating reflector at the other end. The reflectivity of the microsphere is modulated by changing the gap between the microsphere and the fiber taper. Active Q switching is realized by oscillating the microsphere in and out of contact with the taper. Using this novel technique, we have obtained giant pulses (maximum peak power ˜102W, duration ˜160ns) at a low pump-power threshold (˜3mW).
Oxygen sensing glucose biosensors based on alginate nano-micro systems
NASA Astrophysics Data System (ADS)
Chaudhari, Rashmi; Joshi, Abhijeet; Srivastava, Rohit
2014-04-01
Clinically glucose monitoring in diabetes management is done by point-measurement. However, an accurate, continuous glucose monitoring, and minimally invasive method is desirable. The research aims at developing fluorescence-mediated glucose detecting biosensors based on near-infrared radiation (NIR) oxygen sensitive dyes. Biosensors based on Glucose oxidase (GOx)-Rudpp loaded alginate microspheres (GRAM) and GOx-Platinum-octaethylporphyrin (PtOEP)-PLAalginate microsphere system (GPAM) were developed using air-driven atomization and characterized using optical microscopy, CLSM, fluorescence spectro-photometry etc. Biosensing studies were performed by exposing standard solutions of glucose. Uniform sized GRAM and GPAM with size 50+/-10μm were formed using atomization. CLSM imaging of biosensors suggests that Rudpp and PtOEP nanoparticles are uniformly distributed in alginate microspheres. The GRAM and GPAM showed a good regression constant of 0.974 and of 0.9648 over a range of 0-10 mM of glucose with a high sensitivity of 3.349%/mM (625 nm) and 2.38%/mM (645 nm) at 10 mM of glucose for GRAM and GPAM biosensor. GRAM and GPAM biosensors show great potential in development of an accurate and minimally invasive glucose biosensor. NIR dye based assays can aid sensitive, minimally-invasive and interference-free detection of glucose in diabetic patients.
Development of a Luminex based competitive immunoassay for 2,4,6-trinitrotoluene (TNT).
Anderson, George P; Lamar, Jacqueline D; Charles, Paul T
2007-04-15
Previously, a displacement immunoassay for 2,4,6-trinitrotoluene (TNT) was demonstrated using the Luminex 100. The work presented utilized this same specialized flow cytometer to demonstrate a highly sensitive and rapid competitive immunoassay for TNT. This required a TNT analog to be attached to the microsphere surface. Various linkers were evaluated; bovine serum albumin provided over 3 times more binding sites in comparison to various shorter diamine linkers. For this assay TNB-coated microspheres were added to samples; then biotinylated anti-TNT antibody and the reporter molecule, Streptavidin-R-Phycoerythrin, were added. In the absence of TNT, a highly fluorescent complex was formed on the surface of the microsphere. The presence of TNT resulted in dose-dependent decreased fluorescence. Various anti-TNT antibodies were evaluated; Mab 30-1 gave the strongest response, yielding the lowest limit of detection (<1.0 ng/mL) and a dynamic range up to 1 microg/mL. Other factors such as reaction time, cross reactivity to other nitro-compounds, evaluation of acetone extracts of TNT contaminated soils, testing in environmental matrices such as fresh water and seawater were all completed. Finally, a multiplex assay for TNT and three protein toxins was successfully conducted using the competitive format.
Multipurpose Fiber Injected-micro-spherical LIDAR System
NASA Technical Reports Server (NTRS)
Abdelayem, Hossin; Jamison, Tracee
2005-01-01
A technological revolution is occurring in the field of fiber lasers. Over the past two years, the level of power has increased from approx. 100 watts to nearly 1 kilowatt. We are developing a novel fiber laser system, which is a satellite-based LIDAR transmitter of multi-lines. The system is made of a hollow fiber filled with micro-spheres doped with lasing materials. Each sphere has its inherent optical cavity, which makes the system a cavity free and in the same time, emits multi-laser lines for simultaneous multi-task operations. The system is also rugged, compact, lightweight, and durable. Our earlier studies on micro-spheres doped with different laser dyes demonstrated the emission of extremely fine laser lines of less than 3 A line-width, which are of interest for spectroscopic applications, sensing, imaging, and optical communications. Individual dye-doped micro-spheres demonstrated a lasing resonance peaks phenomenon in their fluorescence spectra of linear and nonlinear features that do not exist in the bulk dye solutions. Each individual micro-sphere acts as a laser system with inherent cavity, where the fluorescence line suffers multiple internal reflections within the micro-sphere and gains enough energy to become a laser line. Such resonance peaks are dependent on the sphere's morphology, size, shape, and its refractive index. These resonance peaks are named structural resonance, whispering modes or whispering gallery modes, creeping waves, circumferential waves, surfaces modes, and virtual modes. All of these names refer to the same phenomenon of morphology dependent resonance (MDR), which has already been described and predicted precisely by electromagnetic theory and Lorentz-Mie theory since 1908. The resonance peaks become more obvious when the particle size approaches and exceeds the wavelength of the laser used and the relative index of the particle is greater than that of the surrounding medium. Additional information is included in the original extended abstract.
Individual bioaerosol particle discrimination by multi-photon excited fluorescence.
Kiselev, Denis; Bonacina, Luigi; Wolf, Jean-Pierre
2011-11-21
Femtosecond laser induced multi-photon excited fluorescence (MPEF) from individual airborne particles is tested for the first time for discriminating bioaerosols. The fluorescence spectra, analysed in 32 channels, exhibit a composite character originating from simultaneous two-photon and three-photon excitation at 790 nm. Simulants of bacteria aggregates (clusters of dyed polystyrene microspheres) and different pollen particles (Ragweed, Pecan, Mulberry) are clearly discriminated by their MPEF spectra. This demonstration experiment opens the way to more sophisticated spectroscopic schemes like pump-probe and coherent control. © 2011 Optical Society of America
NASA Astrophysics Data System (ADS)
Mansoor, I.; Liu, Y.; Häfeli, U. O.; Stoeber, B.
2013-08-01
Transdermal drug delivery using microneedles is a technique to potentially replace hypodermic needles for injection of many vaccines and drugs. Fabrication of hollow metallic microneedles so far has been associated with time-consuming steps that restrict batch production of these devices. Here, we are presenting a novel method for making metallic microneedles with any desired height, spacing, and lumen size. In our process, we use solvent casting to coat a mold, which contains an array of pillars, with a conductive polymer composite layer. The conductive layer is then used as a seed layer in a metal electrodeposition process. To characterize the process, the conductivity of the polymer composite with respect to different filler concentrations was investigated. In addition, plasma etching of the polymer was characterized. The electroplating process was also studied further to control the thickness of the microneedle array plate. The strength of the microneedle devices was evaluated through a series of compression tests, while their performance for transdermal drug delivery was tested by injection of 2.28 µm fluorescent microspheres into animal skin. The fabricated metallic microneedles seem appropriate for subcutaneous delivery of drugs and microspheres.
Performance of computer vision in vivo flow cytometry with low fluorescence contrast
Markovic, Stacey; Li, Siyuan; Niedre, Mark
2015-01-01
Abstract. Detection and enumeration of circulating cells in the bloodstream of small animals are important in many areas of preclinical biomedical research, including cancer metastasis, immunology, and reproductive medicine. Optical in vivo flow cytometry (IVFC) represents a class of technologies that allow noninvasive and continuous enumeration of circulating cells without drawing blood samples. We recently developed a technique termed computer vision in vivo flow cytometry (CV-IVFC) that uses a high-sensitivity fluorescence camera and an automated computer vision algorithm to interrogate relatively large circulating blood volumes in the ear of a mouse. We detected circulating cells at concentrations as low as 20 cells/mL. In the present work, we characterized the performance of CV-IVFC with low-contrast imaging conditions with (1) weak cell fluorescent labeling using cell-simulating fluorescent microspheres with varying brightness and (2) high background tissue autofluorescence by varying autofluorescence properties of optical phantoms. Our analysis indicates that CV-IVFC can robustly track and enumerate circulating cells with at least 50% sensitivity even in conditions with two orders of magnitude degraded contrast than our previous in vivo work. These results support the significant potential utility of CV-IVFC in a wide range of in vivo biological models. PMID:25822954
Wei, Yi; Wang, Yuxia; Zhang, Huixia; Zhou, Weiqing; Ma, Guanghui
2016-09-15
A new strategy is developed to prepare porous microspheres with narrow size distribution for peptides controlled release, involving a fabrication of porous microspheres without any porogens followed by a pore closing process. Amphiphilic polymers with different hydrophobic segments (poly(monomethoxypolyethylene glycol-co-d,l-lactide) (mPEG-PLA), poly(monomethoxypolyethylene glycol-co-d,l-lactic-co-glycolic acid) (mPEG-PLGA)) are employed as microspheres matrix to prepare porous microspheres based on a double emulsion-premix membrane emulsification technique combined with a solvent evaporation method. Both microspheres possess narrow size distribution and porous surface, which are mainly caused by (a) hydrophilic polyethylene glycol (PEG) segments absorbing water molecules followed by a water evaporation process and (b) local explosion of microspheres due to fast evaporation of dichloromethane (MC). Importantly, mPEG-PLGA microspheres have a honeycomb like structure while mPEG-PLA microspheres have a solid structure internally, illustrating that the different hydrophobic segments could modulate the affinity between solvent and matrix polymer and influence the phase separation rate of microspheres matrix. Long term release patterns are demonstrated with pore-closed microspheres, which are prepared from mPEG-PLGA microspheres loading salmon calcitonin (SCT). These results suggest that it is potential to construct porous microspheres for drug sustained release using permanent geometric templates as new porogens. Copyright © 2016 Elsevier Inc. All rights reserved.
Thermal response of chalcogenide microsphere resonators
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ahmad, H; Aryanfar, I; Lim, K S
2012-05-31
A chalcogenide microsphere resonator (CMR) used for temperature sensing is proposed and demonstrated. The CMR is fabricated using a simple technique of heating chalcogenide glass and allowing the molten glass to form a microsphere on the waist of a tapered silica fibre. The thermal responses of the CMR is investigated and compared to that of a single-mode-fibre (SMF) based microsphere resonator. It is observed that the CMR sensitivity to ambient temperature changes is 8 times higher than that of the SMF-based microsphere resonator. Heating the chalcogenide microsphere with a laser beam periodically turned on and off shows periodic shifts inmore » the transmission spectrum of the resonator. By injecting an intensity-modulated cw signal through the resonator a thermal relaxation time of 55 ms is estimated.« less
NASA Astrophysics Data System (ADS)
Zhang, Zhaohui; Liu, Li; Li, Hui; Yao, Shouzhuo
2009-09-01
A novel core-shell molecularly imprinting microspheres (MIMs) with trans-resveratrol as the template molecule; acrylamide (AA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as cross-linker, was prepared based on SiO 2 microspheres with surface imprinting technique. These core-shell trans-resveratrol imprinted microspheres were characterized by infrared spectra (IR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and high performance liquid chromatography (HPLC). The results showed that these core-shell imprinted microspheres, which take on perfect spherical shape with average shell thickness of 150 nm, exhibit especially selective recognition for trans-resveratrol. These imprinted microspheres were applied as solid-phase extraction materials for selective extraction of trans-resveratrol from giant knotweed extracting solution successfully.
Trophic level transfer of microplastic: Mytilus edulis (L.) to Carcinus maenas (L.).
Farrell, Paul; Nelson, Kathryn
2013-06-01
This study investigated the trophic transfer of microplastic from mussels to crabs. Mussels (Mytilus edulis) were exposed to 0.5 μm fluorescent polystyrene microspheres, then fed to crabs (Carcinus maenas). Tissue samples were then taken at intervals up to 21 days. The number of microspheres in the haemolymph of the crabs was highest at 24 h (15 033 ml(-1) ± SE 3146), and was almost gone after 21 days (267 ml(-1) ± SE 120). The maximum amount of microspheres in the haemolymph was 0.04% of the amount to which the mussels were exposed. Microspheres were also found in the stomach, hepatopancreas, ovary and gills of the crabs, in decreasing numbers over the trial period. This study is the first to show 'natural' trophic transfer of microplastic, and its translocation to haemolymph and tissues of a crab. This has implications for the health of marine organisms, the wider food web and humans. Copyright © 2013 Elsevier Ltd. All rights reserved.
Computational Spectroscopy of Structured Carbon Nanotube Interfaces for Biochemical Sensing
2010-04-01
oxidized layer of SiCh served as a diffusion barrier to prevent Pt silicide formation and was shown to aide the formation of well dispersed catalytic nano...Three types of fluorescent mi- crospheres (Fluospheres) from Invitrogen were tested. The fluorescent microspheres were prepared in a 10:1 solution with...spectral channels for a given pixel location. Carbon Nanotube Growth on Pt Sample preparation involved cleaving silicon (100) wafers to desired
DOE Office of Scientific and Technical Information (OSTI.GOV)
Chen, S.; Chu, W., E-mail: chuwei65@yahoo.com.cn; Huang, Y.Y.
Graphical abstract: Porous N-doped TiO{sub 2} microspheres were prepared for the first time via plasma technique. The sample exhibited better photocatalytic activity, photoinduced inactivation activity and better electrochemical activity than those of TiO{sub 2} microspheres and P25. Display Omitted Highlights: ► Porous N-doped TiO{sub 2} microspheres were prepared via nitrogen plasma technique. ► Plasma treatment did not affect the porous structure of the TiO{sub 2} microspheres. ► With the plasma treatment, the N contents in the samples increased. ► Their photocatalytic, antibacterial and electrochemical activities were studied. -- Abstract: Nitrogen-doped titanium dioxide (N-doped TiO{sub 2}) microspheres with porous structure weremore » prepared via the nitrogen-assisted glow discharge plasma technique at room temperature for the first time. The samples were characterized by X-ray diffraction, scanning electron microscopy, nitrogen adsorption–desorption measurement, UV–Vis diffuse reflectance spectra, photoluminescence spectroscopy and X-ray photoelectron spectroscopy. The results indicated that the plasma treatment did not affect the porous structure of the TiO{sub 2} microspheres. With the plasma treatment, the N contents in the samples increased. During the photocatalytic degradation of methylene blue under simulative sunlight irradiation, the sample after plasma treatment for 60 min (N-TiO{sub 2}-60) exhibited higher photocatalytic activity than those of the TiO{sub 2} microspheres, P25 and other N-doped TiO{sub 2} microspheres. Furthermore, the N-TiO{sub 2}-60 showed excellent antibacterial activities towards Escherichia coli under visible irradiation. These should be attributed to the enhancement of the visible light region absorption for TiO{sub 2} after N-doping. Electrochemical data demonstrated that the N-doping not only enhanced the electrochemical activity of TiO{sub 2}, but also improved the reversibility of Li insertion/extraction reactions and the rate behavior of TiO{sub 2} during charge–discharge cycles.« less
Percent recovery of low influent concentrations of microorganism surrogates in small sand columns
NASA Astrophysics Data System (ADS)
Stevenson, M. E.; Blaschke, A. P.
2012-04-01
In order to develop a dependable method to calculate the setback distance of a drinking water well from a potential point of microbiological contamination, surrogates are used to perform field tests to avoid using pathogenic micro-organisms. One such surrogate used to model the potential travel time of microbial contamination is synthetic microspheres. The goal of this study is to examine the effect of differing influent colloid concentrations on the percent recovery of microbial surrogates after passing through a soil column. Similar studies have been done to investigate blocking of ideal attachment sites using concentrations between 106 and 1010 particles ml-1. These high concentrations were necessary due to the detection limit of the measuring technique used; however, our measuring technique allows us to test input concentrations ranging from 101 to 106 particles ml-1. These low concentrations are more similar to the concentrations of pathogenic microorganisms present in nature. We have tested the enumeration of 0.5 μm microspheres using a solid-phase cytometer and evaluated their transport in small sand columns. Fluorescent microspheres were purchased for this study with carboxylated surfaces. The soil columns consist of Plexiglas tubes, 30 cm long and 7 cm in diameter, both filled with the same coarse sand. Bromide was used as a conservative tracer, to estimate pore-water velocity and dispersivity, and bromide concentrations were analysed using ion chromatography and bromide probes. Numerical modelling was done using CXTFIT and HYDRUS-1D software programs. The 0.5 μm beads were enumerated in different environmental waters using solid-phase cytometry and compared to counts in sterile water in order to confirm the accuracy of the method. The solid-phase cytometer was able to differentiate the 0.5 μm beads from naturally present autofluorescent particles and bacteria, and therefore, is an appropriate method to enumerate this surrogate.
Chaudhary, Ayesha; Harma, Harri; Hanninen, Pekka; McShane, Michael J; Srivastava, Rohit
2011-08-01
Minimally invasive optical glucose biosensors with increased functional longevity form one of the most promising techniques for continuous glucose monitoring, because of their long-term stability, reversibility, repeatability, specificity, and high sensitivity. They are based on the principle of competitive binding and fluorescence resonance energy transfer. Moving to the near-infrared region of the spectrum has the potential to improve signal throughput for implanted sensors, but requires a change in dye chemistry that could alter response sensitivity, range, and toxicity profiles. The near-infrared dissolved-core alginate microsphere sensors were fabricated by emulsion followed by surface coating by layer-by-layer self-assembly. The particles were characterized for sensor stability, sensor response, and reversibility in deionized water and simulated interstitial fluid. The sensor response to step changes in bulk glucose concentrations was also evaluated under dynamic conditions using a microflow cell unit. Finally, in vitro cytotoxicity assays were performed with L929 mouse fibroblast cell lines to demonstrate preliminary biocompatibility of the sensors. The glucose sensitivity under controlled and dynamic conditions was observed to be 0.86%/mM glucose with an analytical response range of 0-30 mM glucose, covering both the physiological and pathophysiological range. The sensor demonstrated a repeatable, reversible, and reproducible response, with a maximum response time of 120 s. In vitro cytotoxicity assays revealed nearly 95% viability of cells, thereby suggesting that the alginate microsphere sensor system does not exhibit cytotoxicity. The incorporation of near-infrared dyes shows promise in improving sensor response because of their high sensitivity and improved tissue penetration of infrared light. The sensitivity for the sensors was approximately 1.5 times greater than that observed for visible dye sensors, and the new dye chemistry did not significantly alter the biocompatibility of the materials. These findings provide additional support for the potential application of alginate microspheres and similar systems such as "smart-tattoo" glucose sensors.
NASA Astrophysics Data System (ADS)
Lee, Kent; Henze, Dean; Robertson-Anderson, Rae
2013-03-01
Actin is an important cytoskeletal protein involved in cell structure and motility, cancer invasion and metastasis, and muscle contraction. The intricate viscoelastic properties of filamentous actin (F-actin) networks allow for the many dynamic roles of actin, thus warranting investigation. Exploration of this unique stress-strain/strain-rate relationship in complex F-actin networks can also improve biomimetic materials engineering. Here, we use optical tweezers with fluorescence microscopy to study the viscoelastic properties of F-actin networks on the microscopic level. Optically trapped microspheres embedded in various F-actin networks are moved through the network using a nanoprecision piezoelectric stage. The force exerted on the microspheres by the F-actin network and subsequent force relaxation are measured, while a fraction of the filaments in the network are fluorescent-labeled to observe filament deformation in real-time. The dependence of the viscoelastic properties of the network on strain rates and amplitudes as well as F-actin concentration is quantified. This approach provides the much-needed link between induced force and deformation over localized regimes (tens of microns) and down to the single molecule level.
NASA Astrophysics Data System (ADS)
Langford, R. P.; Pillai, S.; Schulze-Makuch, D.; Widmer, K.; Abdel-Fattah, A.; Lerhner, T.
2003-12-01
This study tracks the transport of bromide and microspheres mimicking pathogens in an arid environment. The study site uses the Rio Grande that experiences significant annual fluctuations in both water quantity and quality. The pumping well is 17 m from the stream bank and the water table was 2 m below the stream surface. The aquifer is medium and fine-grained sand comprising two flow units. Observation wells are screened over 1 or 1.5 m intervals. The average hydraulic conductivity was about 2 x 10-3 m/s based on a test analysis, however, the responses indicated that sediment heterogeneities affected the hydraulic behavior. A 427 hour tracer test using bromide and fluorescent microspheres provides initial results that are relevant to the transport of pathogens through the subsurface under riverbank filtration conditions. Bromide was injected into an observation well at the channel margin. Differently colored fluorescent microspheres (0.25nm, 1?m, 6?m and 10?m) were injected into the stream bottom and into two observation wells. Conclusions from the tracer test are: 1) Both bromide and microspheres continued to be observed throughout the 18 days of the experiment. 2) The bromide recovery in the pumping well and in the deeper observation wells showed early and late peaks with a long tails indicating that the geological medium at the field site behaves like a double-porosity medium allowing the tracer to move relatively quickly through the higher conductivity units while being significantly retarded in the low hydraulic conductivity units. 3) Some wells showed consistently higher concentrations of bromide. 4) The 1? micospheres were abundant in the observation wells and allowed tracing of flowpaths. These showed multiple peaks similar to the bromide results. This indicates highly preferential transport paths in the sediment. 5) Microspheres from the three injection sites had distinctly different transport paths and rates. 6) Both bromide and microspheres appeared in the stream soon after injection, moving apparently against an 2-m head difference. 7) The 6 ? and 10 ? microspheres were observed in low concentrations and were episodically detected in the stream and in two widely spaced observation wells. The significance of these results is that: 1) Inorganic microspheres may mimic the episodic occurrence of microorganisms in wells. 2) Even in this relatively homogeneous aquifer, preferential transport within the aquifer results in highly divergent transport paths and rates. Microspheres from one of the injection sites traveled essentially perpendicular to the expected transport direction. 3) Even small variations in the sand grain size can effectively compartmentalize the aquifer. The next steps of this project will include field studies to observe the migration and persistence of selected organisms (E.coli, enterococci, coliphages, cysts, oocysts and enteroviruses) in the pumping well and observation wells under different pumping rates. Continued combined chemical sampling along with the microbial sampling will document the whether changes in water chemistry alter the behavior of the organisms.
[Interaction of protein with charged colloidal particles].
Durdenko, E V; Kuznetsova, S M; Basova, L V; Tikhonenko, S A; Saburova, E A
2011-01-01
The functional state of three proteins of different molecular weight (urease, lactate dehydrogenase, and hemoglobin) in the presence of the linear polyelectrolytes poly(allylamine hydrochloride) (PAA) and sodium poly(styrenesulfonate) (PSS) in the dissolved state and of the same polyelectrolytes bound to the surface of microspheres has been investigated. Microspheres were prepared by consecutive absorption of oppositely charged polyelectrolytes so that the outer layer of the shell was PAA for the acidic protein urease, and PSS for the alkaline proteins LDH and hemoglobin. It was shown that the dissolved polyelectrolyte completely inactivates all three proteins within one minute with a slight difference in the time constant. (By Hb inactivation are conventionally meant changes in the heme environment observed from the spectrum in the Soret band.) In the presence of microspheres, the proteins were adsorbed on their surface; in this case, more than 95% of the activity was retained within two hours. The proportion of the protein adsorbed on microspheres accounted for about 98% for urease, 72% for Hb, and 35% for LDH, as determined from the tryptophan fluorescence data. The interaction of hemoglobin with another type of charged colloidal particles, phospholipid vesicles, leads to the destruction of the tertiary structure of the protein, which made itself evident in the optical absorption spectra in the Soret band, as well as the spectra of tryptophan fluorescence and circular dichroism. In this case, according to circular dichroism, the percentage of alpha-helical structure of Hb was maintained. The differences in the physical and chemical mechanisms of interaction of proteins with these two types of charged colloidal particles that leads to differences in the degree of denaturing effects are discussed.
Gradient changes in porcine renal arterial vascular anatomy and blood flow after cryoablation.
Lagerveld, Brunolf W; van Horssen, Pepijn; Laguna, M Pilar; van den Wijngaard, Jeroen P H M; Siebes, Maria; Wijkstra, Hessel; de la Rosette, Jean J M C H; Spaan, Jos A E
2011-08-01
We quantified temporal changes in vascular structure and blood flow after cryosurgery of the porcine kidney in vivo. We studied 5 groups of 4 kidneys each with a survival time of 20 minutes, 4 hours, 2 days, and 1 and 2 weeks after cryoablation, respectively. Before harvesting the kidneys, fluorescently labeled microspheres were administrated in the descending aorta. After harvest the kidney and its vasculature were casted with fluorescently dyed elastomer, frozen and processed in an imaging cryomicrotome to reveal the 3-dimensional arterial branching structure and microsphere distribution. In regions of interest vessels were segmented by image analysis software and histograms were constructed to reveal the total summed vessel length as a function of diameter. A characteristic diameter of the ablated area was measured. The 20-minute survival group histograms showed a significant shift of the peak to larger diameters (p<0.002), indicating that smaller vessels were destroyed. Microsphere density was decreased to 2% in the ablated region but not in the nonablated border zone, depending on the remaining crater crossing larger vessels. After 2 weeks neither vessels nor microspheres were left in the ablated area, which had shrunk by about 40% in diameter. Study limitations are the lack of histological confirmation and the use of normal rather than cancerous tissue. Larger vessels remain patent just after ablation and transport blood to the border of the ablation crater but perfusion within the crater is halted instantly. Characteristic crater diameter increases initially but decreases thereafter. Destruction of vessels and tissue is complete 2 weeks after cryoablation. Copyright © 2011 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.
Dialysis membrane for separation on microchips
Singh, Anup K [San Francisco, CA; Kirby, Brian J [San Francisco, CA; Shepodd, Timothy J [Livermore, CA
2010-07-13
Laser-induced phase-separation polymerization of a porous acrylate polymer is used for in-situ fabrication of dialysis membranes inside glass microchannels. A shaped 355 nm laser beam is used to produce a porous polymer membrane with a thickness of about 15 .mu.m, which bonds to the glass microchannel and forms a semi-permeable membrane. Differential permeation through a membrane formed with pentaerythritol triacrylate was observed and quantified by comparing the response of the membrane to fluorescein and fluorescently tagging 200 nm latex microspheres. Differential permeation was observed and quantified by comparing the response to rhodamine 560 and lactalbumin protein in a membrane formed with SPE-methylene bisacrylamide. The porous membranes illustrate the capability for the present technique to integrate sample cleanup into chip-based analysis systems.
Controlled release of anticancer drug methotrexate from biodegradable gelatin microspheres.
Narayani, R; Rao, K P
1994-01-01
Biodegradable hydrophilic gelatin microspheres containing the anticancer drug methotrexate (MTX) of different mean particle sizes (1-5, 5-10, and 15-20 microns) were prepared by polymer dispersion technique and crosslinked with glutaraldehyde. The microspheres were uniform, smooth, solid and in the form of free-flowing powder. About 80 per cent of MTX was incorporated in gelatin microspheres of different sizes. The in vitro release of MTX was investigated in two different media, namely simulated gastric and intestinal fluids. The release profiles indicated that gelatin microspheres released MTX in a zero-order fashion for 4-6 days in simulated gastric fluid and for 5-8 days in simulated intestinal fluid. The rate of release of MTX decreased with increase in the particle size of the microspheres. MTX release was faster in gastric fluid when compared to intestinal fluid.
Influence of macroporosity on preferential solute and colloid transport in unsaturated field soils.
Cey, Edwin E; Rudolph, David L; Passmore, Joanna
2009-06-26
Transport of solutes and colloids in soils, particularly those subject to preferential flow along macropores, is important for assessing the vulnerability of shallow groundwater to contamination. The objective of this study was to investigate flow and transport phenomena for dissolved and colloid tracers during large infiltration events in partially saturated, macroporous soils. Controlled tracer infiltration tests were completed at two field sites in southern Ontario. A tension infiltrometer (TI) was used to infiltrate water with dissolved Brilliant Blue FCF dye simultaneously with 3.7 microm and 0.53 microm diameter fluorescent microspheres. Infiltration was conducted under maximum infiltration pressure heads ranging from -5.2 to -0.4 cm. All infiltration test sites were excavated to examine and photograph dye-stained flow patterns, map soil features, and collect samples for microsphere enumeration. Results indicated that preferential transport of dye and microspheres via macropores occurred when maximum pressure heads were greater than -3.0 cm, and the corresponding infiltration rates exceeded 2.0 cm h(-1). Dye and microspheres were detected at depths greater than 70 cm under the highest infiltration rates from both sites. Microsphere concentrations in the top 5-10 cm of soil decreased by more than two orders of magnitude relative to input concentrations, yet remained relatively constant with depth thereafter. There was some evidence for increased retention of the 3.7 microm microspheres relative to the 0.53 microm microspheres, particularly at lower infiltration pressures where straining and attachment mechanisms are most prevalent. Microspheres were observed within dye stained soil matrix surrounding individual macropores, illustrating the significance of capillary pressures in controlling the vertical migration of both tracers in the vicinity of the macropores. Overall, microsphere distributions closely followed the dye patterns, with microsphere concentrations at all depths directly related to the intensity (or concentration) of dye staining. It is concluded that the flow system influenced transport to a much greater degree than differences between dissolved and colloidal species, and hence a dye tracer could serve as a reasonable surrogate for colloid distributions in the vadose zone following individual infiltration events.
Preparation and drug release properties of chitosan/organomodified palygorskite microspheres.
Wu, Jie; Ding, Shijie; Chen, Jing; Zhou, Suqin; Ding, Hongyan
2014-07-01
The novel composite microspheres, based on the hybridization of chitosan (CS) and organomodified palygorskite (OPAL), were prepared by emulsion cross-linking technique and applied as a drug carrier. Palygorskite, a kind of natural one-dimensional clay, was modified with hexadecyl betaine (BS-16) to improve the compatibility and affinity with chitosan matrix, and worked as a perfect micron-filler to enhance drug encapsulation and retard drug migration. The structure of the microspheres was characterized by fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and X-ray diffraction (XRD) techniques. The swelling behavior of the microspheres and the effect of the amount of OPAL and BS-16 on the properties of the drug loading and releasing have been investigated. Compared to the pure chitosan microspheres (CM), the composite one with 20wt% OPAL modified by 20mmol/100g BS-16 possessed the higher encapsulation efficiency and the slower and continuous cumulative release for diclofenac sodium (DS) in phosphate buffer solution (pH 6.8). The study of drug release kinetics in vitro found that the drug release mechanism of the microspheres changed from the simple diffusion-control to diffusion and dissolution-control as the OPAL content in matrix increased from 0 to 20wt%. Copyright © 2014 Elsevier B.V. All rights reserved.
Fabrication of an Optical Fiber Micro-Sphere with a Diameter of Several Tens of Micrometers.
Yu, Huijuan; Huang, Qiangxian; Zhao, Jian
2014-06-25
A new method to fabricate an integrated optical fiber micro-sphere with a diameter within 100 µm, based on the optical fiber tapering technique and the Taguchi method is proposed. Using a 125 µm diameter single-mode (SM) optical fiber, an optical fiber taper with a cone angle is formed with the tapering technique, and the fabrication optimization of a micro-sphere with a diameter of less than 100 µm is achieved using the Taguchi method. The optimum combination of process factors levels is obtained, and the signal-to-noise ratio (SNR) of three quality evaluation parameters and the significance of each process factors influencing them are selected as the two standards. Using the minimum zone method (MZM) to evaluate the quality of the fabricated optical fiber micro-sphere, a three-dimensional (3D) numerical fitting image of its surface profile and the true sphericity are subsequently realized. From the results, an optical fiber micro-sphere with a two-dimensional (2D) diameter less than 80 µm, 2D roundness error less than 0.70 µm, 2D offset distance between the micro-sphere center and the fiber stylus central line less than 0.65 µm, and true sphericity of about 0.5 µm, is fabricated.
Liu, Jianming; Lv, Xin
2014-01-01
The purpose of this study was to develop novel lidocaine microspheres. Microspheres were prepared by the oil-in-water (o/w) emulsion technique using poly(d,l-lactide-co-glycolide acid) (PLGA) for the controlled delivery of lidocaine. The average diameter of lidocaine PLGA microspheres was 2.34 ± 0.3 μm. The poly disperse index was 0.21 ± 0.03, and the zeta potential was +0.34 ± 0.02 mV. The encapsulation efficiency and drug loading of the prepared microspheres were 90.5% ± 4.3% and 11.2% ± 1.4%. In vitro release indicated that the lidocaine microspheres had a well-sustained release efficacy, and in vivo studies showed that the area under the curve of lidocaine in microspheres was 2.02–2.06-fold that of lidocaine injection (p < 0.05). The pharmacodynamics results showed that lidocaine microspheres showed a significant release effect in rats, that the process to achieve efficacy was calm and lasting and that the analgesic effect had a significant dose-dependency. PMID:25268618
NASA Astrophysics Data System (ADS)
Javiya, Curie
Biodegradable poly-(d,l-lactide-co-glycolide) (PLGA) based microspheres are commonly used for numerous clinical applications. PEG is a widely used polymer due to its hydrophilic, biocompatible, and nontoxic nature. In this study, different blends of PLGA/PEG microspheres were prepared using a spray drying technique. The microspheres were spherical with maximum yield found to be 60.3% and average particle size in the range of 2.4 to 3.1 microm. Under the spray drying processing conditions, the polymers showed full miscibility slightly below 15% w/w and partial miscibility up to 20% w/w of PEG in the blended microspheres. At higher temperatures, PLGA and PEG were miscible in all proportions used for the blended microspheres. Blending 10% w/w PEG in PLGA membranes showed significant reduction in attachment of macrophages compared to PLGA membranes. The in-vitro response of macrophage towards the miscible blends of PLGA/PEG microspheres was further characterized. Results showed some reduction in macrophage viability and activation, however, significant effects with PLGA/PEG microspheres were not observed.
Influence of diameter on particle transport in a fractured shale saprolite
Cumbie, D.H.; McKay, L.D.
1999-01-01
Experiments in an undisturbed, saturated column of weathered and fractured shale saprolite using fluorescent carboxylate-coated latex microspheres as tracers indicate that particle diameter plays a major role in controlling transport. In this study the optimum microsphere diameter for transport was approximately 0.5 ??m. Microspheres larger than the optimum size were present in the effluent at lower relative concentrations, apparently because of greater retention due to gravitational settling and/or physical straining. The smaller than optimum microspheres also experienced greater retention, apparently related to their higher rates of diffusion. Faster diffusion can lead to more frequent collisions with, and attachment to, fracture walls and may also lead to movement of particles into zones of relatively immobile pore water in the fractures or in the fine pore structure of the clay-rich matrix between fractures. Dismantling of the soil column and mapping of the distribution of retained microspheres indicated that there was substantial size-segregation of the microspheres between different fractures or in 'channels' within a fracture. Examination of small core samples showed that the smallest microspheres (0.05-0.1 ??m) were present in the fine pores of the matrix at distances of up to 3-4 mm from the nearest fracture, which supports the hypothesis that small particles can be retained by diffusion into the matrix. Calculations of settling velocity and diffusion rate using simple 1D approaches suggest that these processes could both cause significant retention of the larger and smaller particles, respectively, even for the fast advective transport rates (up to 32 m/day) observed during the experiments. Copyright (C) 1999 Elsevier Science B.V.
Micro-heterogeneity of Cellulosic Fiber Biopolymer Prepared from Corn Hulls
USDA-ARS?s Scientific Manuscript database
Z-trim is a zero calorie cellulosic fiber biopolymer produced from corn hulls. The micro-structural heterogeneities of Z-trim biopolymer were investigated by monitoring the thermally driven displacements of well-dispersed micro-spheres via video fluorescence microscopy named multiple-particle track...
Micro-Heterogeneity of Cellulosic Fiber Biopolymer Prepared from Corn Hulls
USDA-ARS?s Scientific Manuscript database
Z-trim is a zero calorie cellulosic fiber biopolymer produced from corn hulls. The micro-structural heterogeneities of Z-trim biopolymer were investigated by monitoring the thermally driven displacements of well-dispersed micro-spheres via video fluorescence microscopy named multiple-particle track...
Zhang, Ding Sheng-Zi; Jiang, Yang; Wei, Dan; Wei, Xunbin; Xu, Hong; Gu, Hongchen
2018-06-21
With the increasing demands for high-throughput multiplexed bioassays, quantum dot (QD)-encoded microbeads as biocarriers for various bioreactions have attracted considerable attention. However, three key requirements for these biocarriers are still longstanding issues: a stable fluorescence intensity, a large encoding capacity and abundant surface functional groups. Here, a novel one-pot strategy is developed, generating functionalized QD-encoded microspheres with a strong fluorescence intensity and optical stability. With poly(styrene-co-maleic anhydride) (PSMA) molecules as mediators, the encapsulation of QDs and carboxylation of the bead surface are integrated together, greatly improving the preparation efficiency and guaranteeing their potential application in biodetection. Moreover, the mechanism for preparing QD-doped beads is further proposed, which helps to precisely manipulate the preparation process and accurately encode the beads. Through this approach, a single- and dual-color barcode library of QD-encoded microspheres has been successfully established, which demonstrates their great potential in suspension arrays.
Getting the most from microfluidic platforms for biomedical applications (Conference Presentation)
NASA Astrophysics Data System (ADS)
Shen, Amy
2016-03-01
Microfluidics has emerged in recent years as a versatile method of manipulating fluids at small length-scales, and in particular, for generating and manipulating micron size droplets with controllable size and functionality. For example, many research groups developed microfluidics devices for cell encapsulation, and synthesizing functionalized polymer microspheres and inorganic nanoparticles with precise control over their shapes and sizes. In this talk, I will showcase 2 microfluidic platforms to highlight their versatility and potential biomedical applications. (1) Droplet microfluidic platforms (a) A droplet microfluidics method to fabricate alginate microspheres while simultaneously immobilizing anti-Mycobacterium tuberculosis complex IgY and anti-Escherichia coli IgG antibodies primarily on the porous alginate carriers for specific binding and binding affinity tests. The binding affinity of antibodies is directly measured by fluorescence intensity of stained target bacteria on the microspheres. We demonstrate that the functionalized alginate microspheres yield specificity comparable with an enzyme-linked immunosorbent assay. We can easily modify the size and shape of alginate microspheres, and increase the concentration of functionalized alginate microspheres to further enhance binding kinetics and enable multiplexing. (b) A novel droplet microfluidics method to image oxygen in single islets (pancreatic cells) for glucose sensing. Individual islets and a fluorescent oxygen-sensitive dye were encased within a thin alginate polymer microcapsule for insulin secretion monitoring. The sensing system operated similarly from 2-48 hours following encapsulation, and viability and function of the islets were not significantly affected by the encapsulation process. This approach should be applicable to other cell types and dyes sensitive to other biologically important molecules. (2) A microfluidic chamber to perform uniform electric field stimulation in circular shaped culturewares A 3D computer-aided designed (CAD) polymeric insert is designed and retrofitted to circular shaped culturewares in an integrated microfluidic electrical stimulation platform to generate uniform EF with higher cell yields. In particular, NIH/3T3 mouse embryonic fibroblast cells are used to validate the performance of the 3D designed Poly(methyl methacrylate) (PMMA) inserts in a circular-shaped 6-well plate. The CAD based inserts can be easily scaled up to further increase effective stimulation area percentages, and also be implemented in commercially available culturewares for a wide variety of EF-related research such as EF-cell interaction and tissue regeneration studies.
Terada, Takatoshi; Ohtsubo, Toshiro; Iwao, Yasunori; Noguchi, Shuji; Itai, Shigeru
2017-01-01
The purpose of this study was to develop a deeper understanding of the key physicochemical parameters involved in the release profiles of microsphere-encapsulated agrochemicals at different temperatures. Microspheres consisting of different polyurethanes (PUs) were prepared using our previously reported solventless microencapsulation technique. Notably, these microspheres exhibited considerable differences in their thermodynamic characteristics, including their glass transition temperature (T g ), extrapolated onset temperature (T o ) and extrapolated end temperature (T e ). At test temperatures below the T o of the PU, only 5-10% of the agrochemical was rapidly released from the microspheres within 1 d, and none was released thereafter. However, at test temperatures above the T o of the PU, the rate of agrochemical release gradually increased with increasing temperatures, and the rate of release from the microspheres was dependent on the composition of the PU. Taken together, these results show that the release profiles of the microspheres were dependent on their thermodynamic characteristics and changes in their PU composition.
Production of cerium dioxide microspheres by an internal gelation sol–gel method
DOE Office of Scientific and Technical Information (OSTI.GOV)
Katalenich, Jeffrey A.
An internal gelation sol-gel technique was used to prepare cerium dioxide microspheres with uniform diameters near 100 µm. In this process, chilled aqueous solutions containing cerium, hexamethylenetetramine (HMTA), and urea are transformed into a solid gel by heat addition and are subsequently washed, dried, and sintered to produce pure cerium dioxide. Cerous nitrate and ceric ammonium nitrate solutions were compared for their usefulness in microsphere production. Gelation experiments were performed with both cerous nitrate and ceric ammonium nitrate to determine desirable concentrations of cerium, HMTA, and urea in feed solutions as well as the necessary quantity of ammonium hydroxide addedmore » to cerium solutions. Analysis of the pH before and after sample gelation was found to provide a quantitative metric for optimal parameter selection along with subjective evaluations of gel qualities. The time necessary for chilled solutions to gel upon inserting into a hot water bath was determined for samples with a variety of parameters and also used to determine desirable formulations for microsphere production. A technique for choosing the optimal mixture of ceric ammonium nitrate, HMTA, and urea was determined using gelation experiments and used to produce microspheres by dispersion of the feed solution into heated silicone oil. Gelled spheres were washed to remove excess reactants and reaction products before being dried and sintered. X-ray diffraction of air-dried microspheres, sintered microspheres, and commercial CeO 2 powders indicated that air-dried and sintered spheres were pure CeO 2.« less
Crestani, Sandra; Leitolis, Amanda; Lima, Lucianna Freitas Oliveira; Krieger, Marco A; Foti, Leonardo
2016-08-01
Diverse techniques have been developed to analyze antibody-mediated responses to infections. However, the most common tests, i.e., enzyme-linked immunosorbent assays, require separate reactions for each antigen and consequently necessitate large sample volumes. Luminex technology allows the detection of multiple antibodies in a single experiment, but nonspecific binding can impair the results. Therefore, we examined the use of Escherichia coli lysates to reduce nonspecific binding and improve the results of liquid microarrays based on Luminex technology. Anti-bacteria antibodies were detected in human serum samples, as evidenced by high median fluorescence intensity (MFI) in assays performed with paramagnetic microspheres coupled with E. coli lysates. Moreover, the addition of an E. coli lysate as a blocker reduced the nonspecific binding of antigens produced by E. coli in a concentration-dependent manner. Tris-HCl reduced MFI values in negative samples, but did not affect MFI for positive samples. For microspheres coupled with different antigens, an E. coli lysate blocker significantly improved the fluorescence signals from positive samples. The addition of Tris-HCl and the E. coli lysate induced antigen-specific differences in MFI. This combination of the E. coli lysate blocker and Tris-HCl yielded a statistically significant improvement in MFI in the assays for Chagas disease and hepatitis C virus samples. However, for the Treponema pallidum p47 antigen improvement in MFI was only observed for the preparation with the E. coli blocker at a concentration of 3%. In conclusion, the addition of an E. coli lysate and Tris-HCl to the microarray assay reduced the nonspecific binding of human anti-bacteria antibodies and, therefore, increased the specific MFI. Copyright © 2016 Elsevier B.V. All rights reserved.
Hebert, Benedict; Costantino, Santiago; Wiseman, Paul W
2005-05-01
We introduce a new extension of image correlation spectroscopy (ICS) and image cross-correlation spectroscopy (ICCS) that relies on complete analysis of both the temporal and spatial correlation lags for intensity fluctuations from a laser-scanning microscopy image series. This new approach allows measurement of both diffusion coefficients and velocity vectors (magnitude and direction) for fluorescently labeled membrane proteins in living cells through monitoring of the time evolution of the full space-time correlation function. By using filtering in Fourier space to remove frequencies associated with immobile components, we are able to measure the protein transport even in the presence of a large fraction (>90%) of immobile species. We present the background theory, computer simulations, and analysis of measurements on fluorescent microspheres to demonstrate proof of principle, capabilities, and limitations of the method. We demonstrate mapping of flow vectors for mixed samples containing fluorescent microspheres with different emission wavelengths using space time image cross-correlation. We also present results from two-photon laser-scanning microscopy studies of alpha-actinin/enhanced green fluorescent protein fusion constructs at the basal membrane of living CHO cells. Using space-time image correlation spectroscopy (STICS), we are able to measure protein fluxes with magnitudes of mum/min from retracting lamellar regions and protrusions for adherent cells. We also demonstrate the measurement of correlated directed flows (magnitudes of mum/min) and diffusion of interacting alpha5 integrin/enhanced cyan fluorescent protein and alpha-actinin/enhanced yellow fluorescent protein within living CHO cells. The STICS method permits us to generate complete transport maps of proteins within subregions of the basal membrane even if the protein concentration is too high to perform single particle tracking measurements.
NASA Astrophysics Data System (ADS)
Latina, M. A.; Kobsa, P. H.; Rakestraw, S. L.; Crean, E. A.; Hasan, T.; Yarmush, M. L.
1989-03-01
We have investigated a novel and efficient delivery system utilizing photosensitizer-coupled-latex microspheres to photochemically target and kill phagocytic trabecular meshwork (TM) cells. TM cells are the most actively phagocytic cells within the anterior chamber of the eye and are located within an optically accessible discrete band. This delivery system, along with the property of cell photocytosis, will achieve double selectivity by combining preferential localization of the photosensitizer to the target cells with spatial localization of illumination on the target cells. All experiments were performed with preconfluent bovine TM cells, 3rd to 4th passage, plated in 15 mm wells. Chlorin e6 monoethylene diamine monoamide was conjugated to the surface of 1.0 Am MX Duke Scientific fluorescent latex microspheres. Spectroscopic analysis revealed an average of 1.3 x 10 -17 moles of chlorin e6 per microsphere. TM cells were incubated for 18 hours with 5 x 10 7 microspheres/ml in MEM with 10% FCS, washed with MEM, and irradiated through fresh media using an argon-pumped dye laser emitting .2 W at 660 nm. A dose-survival study indicated that energy doses of 10 J/cm2 or greater resulted in greater than 95% cell death as determined by ethidium bromide exclusion. Cell death could be demonstrated as early as 4 hours post-irradiation. TM cells incubated with a solution of chlorin e6 at a concentration equal to that conjugated to the microspheres showed no cell death. Unirradiated controls also showed no cell death.
Alginate microspheres obtained by the spray drying technique as mucoadhesive carriers of ranitidine.
Szekalska, Marta; Amelian, Aleksandra; Winnicka, Katarzyna
2015-03-01
The present study is aimed at formulation of alginate (ALG) microspheres with ranitidine (RNT) by the spray drying method. Obtained microspheres were characterized for particle size, surface morphology, entrapment efficiency, drug loading, in vitro drug release and zeta potential. Mucoadhesive properties were examined by a texture analyser and three types of adhesive layers--gelatine discs, mucin gel and porcine stomach mucosa. Microspheres showed a smooth surface with narrow particle size distribution and RNT loading of up to 70.9%. All formulations possessed mucoadhesive properties and exhibited prolonged drug release according to the first-order kinetics. DSC reports showed that there was no interaction between RNT and ALG. Designed microspheres can be considered potential carriers of ranitidine with prolonged residence time in the stomach.
A novel method for producing microspheres with semipermeable polymer membranes
NASA Technical Reports Server (NTRS)
Lin, K. C.; Wang, Taylor G.
1992-01-01
A new and systematic approach for producing polymer microspheres has been demonstrated. The membrane of the microsphere is formed by immersing the polyanionic droplet into a collapsing annular sheet, which is made of another polycation polymer solution. This method minimizes the impact force during the time when the chemical reaction takes place, hence eliminating the shortcomings of the current encapsulation techniques. The results of this study show the feasibility of this method for mass production of microcapsules.
van Manen, Henk-Jan; van Apeldoorn, Aart A; Verrijk, Ruud; van Blitterswijk, Clemens A; Otto, Cees
2007-01-01
Micro- and nanospheres composed of biodegradable polymers show promise as versatile devices for the controlled delivery of biopharmaceuticals. Whereas important properties such as drug release profiles, biocompatibility, and (bio)degradability have been determined for many types of biodegradable particles, information about particle degradation inside phagocytic cells is usually lacking. Here, we report the use of confocal Raman microscopy to obtain chemical information about cross-linked dextran hydrogel microspheres and amphiphilic poly(ethylene glycol)-terephthalate/poly(butylene terephthalate) (PEGT/PBT) microspheres inside RAW 264.7 macrophage phagosomes. Using quantitative Raman microspectroscopy, we show that the dextran concentration inside phagocytosed dextran microspheres decreases with cell incubation time. In contrast to dextran microspheres, we did not observe PEGT/PBT microsphere degradation after 1 week of internalization by macrophages, confirming previous studies showing that dextran microsphere degradation proceeds faster than PEGT/PBT degradation. Raman microscopy further showed the conversion of macrophages to lipid-laden foam cells upon prolonged incubation with both types of microspheres, suggesting that a cellular inflammatory response is induced by these biomaterials in cell culture. Our results exemplify the power of Raman microscopy to characterize microsphere degradation in cells and offer exciting prospects for this technique as a noninvasive, label-free optical tool in biomaterials histology and tissue engineering. PMID:17722552
Setia, Anupama; Kansal, Sahil; Goyal, Naveen
2013-07-01
Microspheres constitute an important part of oral drug delivery system by virtue of their small size and efficient carrier capacity. However, the success of these microspheres is limited due to their short residence time at the site of absorption. The objective of the present study was to formulate and systematically evaluate in vitro performance of enteric coated mucoadhesive microspheres of duloxetine hydrochloride (DLX), an acid labile drug. DLX microspheres were prepared by simple emulsification phase separation technique using chitosan as carrier and glutaraldehyde as a cross-linking agent. Microspheres prepared were coated with eudragit L-100 using an oil-in-oil solvent evaporation method. Eudragit L-100was used as enteric coating polymer with the aim to release the drug in small intestine The microspheres prepared were characterized by particle size, entrapment efficiency, swelling index (SI), mucoadhesion time, in vitro drug release and surface morphology. A 3(2) full factorial design was employed to study the effect of independent variables polymer-to-drug ratio (X1) and stirring speed (X2) on dependent variables, particle size, entrapment efficiency, SI, in vitro mucoadhesion and drug release up to 24 h (t24). Microspheres formed were discrete, spherical and free flowing. The microspheres exhibited good mucoadhesive property and also showed high percentage entrapment efficiency. The microspheres were able to sustain the drug release up to 24 h. Thus, the prepared enteric coated mucoadhesive microspheres may prove to be a potential controlled release formulation of DLX for oral administration.
Use of stable isotope-labeled Escherichia coli as a tracer in karst aquifers
USDA-ARS?s Scientific Manuscript database
Bacterial contamination of karst aquifers is a large concern across the globe, yet bacterial transport in karst aquifers is not currently well understood. Groundwater tracers typically used in karst systems include fluorescent dyes and latex microspheres. Not only can these tracers can be cost-prohi...
USDA-ARS?s Scientific Manuscript database
Nutrim-10 is a newly developed food product containing the dietary of soluble fiber ß-glucan. The micro-structural heterogeneities of Nutrim-10 suspensions were investigated by monitoring the thermally driven displacements of well-dispersed microspheres via video fluorescence microscopy. By comparin...
NASA Astrophysics Data System (ADS)
Mazlan, Siti Zulaikha; Hanifah, Sharina Abu
2014-09-01
Immobilization of laccase on the modified copolymer methacrylate-acrylate microspheres was studied. A poly (glycidyl methacrylate-co-n-butyl acrylate) microsphere consists of epoxy groups were synthesized using suspension photocuring technique. The epoxy group in poly (GMA-nBA) microspheres were converted into amino groups with aldehyde group. Laccase immobilization is based on having the amino groups on the enzyme surface and aldehyde group on the microspheres via covalent binding. Fourier transform infrared spectroscopy (FT-IR) analysis proved the successful surface modification on microspheres. The FTIR spectrum shows the characteristic peaks at 1646 cm-1 assigned to the conformation of the polymerization that took place between monomer GMA and nBA respectively. In addition, after modification, FTIR peaks that assigned to the epoxy ring (844 cm-1 and 904 cm-1) were decreased. The results obtained from FTIR method signify good agreement with the epoxy content method. Hence, the activity of the laccase-immobilized microspheres increased upon increasing the epoxy content. Furthermore, poly (GMA-nBA) exhibited uniform microspheres with below 2 μm surface. Immobilized enzyme showed a broader pH profile and higher temperature compared native enzyme.
MO-A-BRD-00: Current Trends in Y90-Microsphere Therapy: Delivery and Dosimetry
DOE Office of Scientific and Technical Information (OSTI.GOV)
NONE
2015-06-15
Yttrium-90 (Y90) microsphere therapy, a form of radiation therapy, is an increasingly popular option for care of patients with liver metastases or unresectable hepatocellular carcinoma. The therapy directly delivers Y90 microspheres via the hepatic artery to disease sites. Following delivery, a vast majority of microspheres preferentially lodge in the capillary vessels due to their embolic size and targeted trans-arterial delivery – depositing up to 90% of its energy in the first 5 mm of tissue. There have been a number of advances in tomographic imaging within both interventional radiology and nuclear medicine that has advanced therapy planning techniques. Quantitative imagingmore » of Y90 microsphere distribution post-therapy has also seen innovations that have led to improvements in tumor dosimetry and characterization of tumor response. A review of current trends and recent innovation in Y90 microsphere therapies will be presented. Learning Objectives: To present the imaging requirements for Y90 microsphere therapy planning To explain the standard dosimetry models used in Y90 microsphere therapy planning To report on advances in imaging for therapy planning and posttherapy assessment of tumor dosimetry and response.« less
Wang, Aiping; Liang, Rongcai; Liu, Wanhui; Sha, Chunjie; Li, Youxin; Sun, Kaoxiang
2016-01-01
The initial burst release is a major obstacle to the development of microsphere-formulated drug products. To investigate the influence of palmitic acid on the characteristics and release profiles of rotigotine-loaded poly(d,l-lactide-co-glycolide) microspheres. Rotigotine-loaded microspheres (RMS) were prepared using the oil-in-water emulsion solvent evaporation technique. The in vitro characteristics of the RMS were evaluated with scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and a particle size analyzer. The in vitro drug release and in vivo pharmacokinetics of the RMS were investigated. The SEM results showed that the addition of palmitic acid changed the surface morphology of the microspheres from smooth to dimpled and then to non-smooth as the palmitic acid content increased. DSC revealed the existence of molecularly dispersed forms of palmitic acid in the microspheres. The in vitro and in vivo release profiles indicated that the addition of 5% and 8% palmitic acid significantly decreased the burst release of rotigotine from the microspheres, and the late-stage release was delayed as the palmitic acid content increased across the investigated range (5-15%). The addition of palmitic acid to the microspheres significantly affects the release profile of rotigotine from RMS.
A short term quality control tool for biodegradable microspheres.
D'Souza, Susan; Faraj, Jabar A; Dorati, Rossella; DeLuca, Patrick P
2014-06-01
Accelerated in vitro release testing methodology has been developed as an indicator of product performance to be used as a discriminatory quality control (QC) technique for the release of clinical and commercial batches of biodegradable microspheres. While product performance of biodegradable microspheres can be verified by in vivo and/or in vitro experiments, such evaluation can be particularly challenging because of slow polymer degradation, resulting in extended study times, labor, and expense. Three batches of Leuprolide poly(lactic-co-glycolic acid) (PLGA) microspheres having varying morphology (process variants having different particle size and specific surface area) were manufactured by the solvent extraction/evaporation technique. Tests involving in vitro release, polymer degradation and hydration of the microspheres were performed on the three batches at 55°C. In vitro peptide release at 55°C was analyzed using a previously derived modification of the Weibull function termed the modified Weibull equation (MWE). Experimental observations and data analysis confirm excellent reproducibility studies within and between batches of the microsphere formulations demonstrating the predictability of the accelerated experiments at 55°C. The accelerated test method was also successfully able to distinguish the in vitro product performance between the three batches having varying morphology (process variants), indicating that it is a suitable QC tool to discriminate product or process variants in clinical or commercial batches of microspheres. Additionally, data analysis utilized the MWE to further quantify the differences obtained from the accelerated in vitro product performance test between process variants, thereby enhancing the discriminatory power of the accelerated methodology at 55°C.
Metronidazole loaded pectin microspheres for colon targeting.
Vaidya, Ankur; Jain, Aviral; Khare, Piush; Agrawal, Ram K; Jain, Sanjay K
2009-11-01
A multiparticulate system having pH-sensitive property and specific enzyme biodegradability for colon-targeted delivery of metronidazole was developed. Pectin microspheres were prepared using emulsion-dehydration technique. These microspheres were coated with Eudragit(R) S-100 using oil-in-oil solvent evaporation method. The SEM was used to characterize the surface of these microspheres and a distinct coating over microspheres could be seen. The in vitro drug release studies exhibited no drug release at gastric pH, however continuous release of drug was observed from the formulation at colonic pH. Further, the release of drug from formulation was found to be higher in the presence of rat caecal contents, indicating the effect of colonic enzymes on the pectin microspheres. The in vivo studies were also performed by assessing the drug concentration in various parts of the GIT at different time intervals which exhibited the potentiality of formulation for colon targeting. Hence, it can be concluded that Eudragit coated pectin microspheres can be used for the colon specific delivery of drug. (c) 2009 Wiley-Liss, Inc. and the American Pharmacists Association
MO-A-BRD-02: Physics Perspective
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kappadath, S.
2015-06-15
Yttrium-90 (Y90) microsphere therapy, a form of radiation therapy, is an increasingly popular option for care of patients with liver metastases or unresectable hepatocellular carcinoma. The therapy directly delivers Y90 microspheres via the hepatic artery to disease sites. Following delivery, a vast majority of microspheres preferentially lodge in the capillary vessels due to their embolic size and targeted trans-arterial delivery – depositing up to 90% of its energy in the first 5 mm of tissue. There have been a number of advances in tomographic imaging within both interventional radiology and nuclear medicine that has advanced therapy planning techniques. Quantitative imagingmore » of Y90 microsphere distribution post-therapy has also seen innovations that have led to improvements in tumor dosimetry and characterization of tumor response. A review of current trends and recent innovation in Y90 microsphere therapies will be presented. Learning Objectives: To present the imaging requirements for Y90 microsphere therapy planning To explain the standard dosimetry models used in Y90 microsphere therapy planning To report on advances in imaging for therapy planning and posttherapy assessment of tumor dosimetry and response.« less
MO-A-BRD-01: Clinical Perspective
DOE Office of Scientific and Technical Information (OSTI.GOV)
Mahvash, A.
2015-06-15
Yttrium-90 (Y90) microsphere therapy, a form of radiation therapy, is an increasingly popular option for care of patients with liver metastases or unresectable hepatocellular carcinoma. The therapy directly delivers Y90 microspheres via the hepatic artery to disease sites. Following delivery, a vast majority of microspheres preferentially lodge in the capillary vessels due to their embolic size and targeted trans-arterial delivery – depositing up to 90% of its energy in the first 5 mm of tissue. There have been a number of advances in tomographic imaging within both interventional radiology and nuclear medicine that has advanced therapy planning techniques. Quantitative imagingmore » of Y90 microsphere distribution post-therapy has also seen innovations that have led to improvements in tumor dosimetry and characterization of tumor response. A review of current trends and recent innovation in Y90 microsphere therapies will be presented. Learning Objectives: To present the imaging requirements for Y90 microsphere therapy planning To explain the standard dosimetry models used in Y90 microsphere therapy planning To report on advances in imaging for therapy planning and posttherapy assessment of tumor dosimetry and response.« less
Method for dialysis on microchips using thin porous polymer membrane
Singh, Anup K [San Francisco, CA; Kirby, Brian J [San Francisco, CA; Shepodd, Timothy J [Livermore, CA
2009-05-19
Laser-induced phase-separation polymerization of a porous acrylate polymer is used for in-situ fabrication of dialysis membranes inside glass microchannels. A shaped 355 nm laser beam is used to produce a porous polymer membrane with a thickness of about 15 .mu.m, which bonds to the glass microchannel and forms a semi-permeable membrane. Differential permeation through a membrane formed with pentaerythritol triacrylate was observed and quantified by comparing the response of the membrane to fluorescein and fluorescently tagging 200 nm latex microspheres. Differential permeation was observed and quantified by comparing the response to rhodamine 560 and lactalbumin protein in a membrane formed with SPE-methylene bisacrylamide. The porous membranes illustrate the capability for the present technique to integrate sample cleanup into chip-based analysis systems.
Dialysis on microchips using thin porous polymer membranes
Singh, Anup K.; Kirby, Brian J.; Shepodd, Timothy J.
2007-09-04
Laser-induced phase-separation polymerization of a porous acrylate polymer is used for in-situ fabrication of dialysis membranes inside glass microchannels. A shaped 355 nm laser beam is used to produce a porous polymer membrane with a thickness of about 15 .mu.m, which bonds to the glass microchannel and form a semi-permeable membrane. Differential permeation through a membrane formed with pentaerythritol triacrylate was observed and quantified by comparing the response of the membrane to fluorescein and fluorescently tagging 200 nm latex microspheres. Differential permeation was observed and quantified by comparing the response to rhodamine 560 and lactalbumin protein in a membrane formed with SPE-methylene bisacrylamide. The porous membranes illustrate the capability for the present technique to integrate sample cleanup into chip-based analysis systems.
SEM visualization of glycosylated surface molecules using lectin-coated microspheres
NASA Technical Reports Server (NTRS)
Duke, J.; Janer, L.; Campbell, M.
1985-01-01
There are several techniques currently used to localize glycosylated surface molecules by scanning electron microscopy (Grinnell, 1980; Molday, 1976; Linthicum and Sell, 1975; Nicolson, 1974; Lo Buglio, et al, 1972). A simple and rapid method, using a modification of Grinnell's technique is reported here. Essentially, microspheres coated with Concavalin A are used to bind to glycosylated regions of the palatal shelf epithelium and are visualized in the scanning electron microscope (SEM).
Puchkov, Evgeny O
2010-06-01
In the vacuoles of Saccharomyces cerevisiae yeast cells, vividly moving insoluble polyphosphate complexes (IPCs) <1 microm size, stainable by a fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI), may appear under some growth conditions. The aim of this study was to quantitatively characterize the movement of the IPCs and to evaluate the viscosity in the vacuoles using the obtained data. Studies were conducted on S. cerevisiae cells stained by DAPI and fluorescein isothyocyanate-labelled latex microspheres, using fluorescence microscopy combined with computer image analysis (ImageJ software, NIH, USA). IPC movement was photorecorded and shown to be Brownian motion. On latex microspheres, a methodology was developed for measuring a fluorescing particle's two-dimensional (2D) displacements and its size. In four yeast cells, the 2D displacements and sizes of the IPCs were evaluated. Apparent viscosity values in the vacuoles of the cells, computed by the Einstein-Smoluchowski equation using the obtained data, were found to be 2.16 +/- 0.60, 2.52 +/- 0.63, 3.32 +/- 0.9 and 11.3 +/- 1.7 cP. The first three viscosity values correspond to 30-40% glycerol solutions. The viscosity value of 11.3 +/- 1.7 cP was supposed to be an overestimation, caused by the peculiarities of the vacuole structure and/or volume in this particular cell. This conclusion was supported by the particular quality of the Brownian motion trajectories set in this cell as compared to the other three cells.
2011-03-21
produced were also labeled with FITC using a modification of a previously described technique [22]. PEI- coated microsphere (30 mg/mL) were mixed with...surface after 4 h of mixing (Fig. 5A), while PEI-coated microspheres were well- dispersed and immobilized onto the HAp surface (Fig. 5B). 3.3. Properties of...Erickson, L. Ren, D. Li, Zeta-potential measurement using the smoluchowski equation and the slope of the current-time relationship in electroosmotic flow
Optical diffraction by ordered 2D arrays of silica microspheres
NASA Astrophysics Data System (ADS)
Shcherbakov, A. A.; Shavdina, O.; Tishchenko, A. V.; Veillas, C.; Verrier, I.; Dellea, O.; Jourlin, Y.
2017-03-01
The article presents experimental and theoretical studies of angular dependent diffraction properties of 2D monolayer arrays of silica microspheres. High-quality large area defect-free monolayers of 1 μm diameter silica microspheres were deposited by the Langmuir-Blodgett technique under an accurate optical control. Measured angular dependencies of zeroth and one of the first order diffraction efficiencies produced by deposited samples were simulated by the rigorous Generalized Source Method taking into account particle size dispersion and lattice nonideality.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Huang, Yantang, E-mail: g@fzu.edu.cn; Huang, Yu; Zhang, Peijin
2014-02-15
An experimental investigation on ultralow threshold laser and blue shift cooperative luminescence (CL) in a Yb{sup 3+} doped silica microsphere (YDSM) with continuous-wave 976 nm laser diode pumping is reported. The experimental results show that the YDSM emits laser oscillation with ultralow threshold of 2.62 μW, and the laser spectrum is modulated by the microsphere morphology characteristics. In addition, blue emission of YDSM is also observed with the increase of pump power, which is supposed to be generated by CL of excited Yb ion-pairs with the absorption of 976 nm photons and Si-O vibration phonons, and the process is explainedmore » with an energy level diagram. This property of the blue shift CL with phonons absorption in the Yb{sup 3+}doped microcavity makes it attractive for the application of laser cooling based on anti-Stokes fluorescence emission, if the Yb{sup 3+}doped microcavity made from with low phonon energy host materials.« less
Bagnol, D; Jule, Y; Kirchner, G; Cupo, A; Roman, C
1993-02-01
Retrograde tracing with rhodamine fluorescent microspheres combined with fluorescein immunolabelling of methionine-enkephalin showed the presence of enkephalin-like material in neurons of the inferior mesenteric ganglion (sympathetic prevertebral ganglion) projecting to the distal colon in cat. Two weeks after injecting the microspheres into the wall of the distal colon, the inferior mesenteric ganglion was dissected out and incubated for 24 hours in a colchicine-containing culture medium in order to facilitate the detection of enkephalins in the soma of ganglion neurons. It was observed that retrogradely labelled ganglion cells contained enkephalin-like immunoreactive material. These ganglion cells corresponded to enkephalin-like postganglionic neurons, the terminals of which were located inside the wall of the distal colon. These enkephalin-like neurons were numerous and scattered throughout the ganglion. Sometimes enkephalin-like immunoreactive fibers, probably originating from spinal preganglionic neurons, ran close to immunoreactive and non-immunoreactive retrogradely labelled ganglion cells. This suggests that enkephalin-like immunoreactive fibers may make synaptic connections with enkephalin-like and non-enkephalin-like postganglionic neurons projecting to the distal colon. The present study establishes for the first time the existence of an enkephalin-like postganglionic pathway to the digestive tract originating from a sympathetic prevertebral ganglion. This finding indicates that the enkephalinergic innervation of the cat digestive tract may have at least two possible sources: (i) the sympathetic prevertebral ganglia; and (ii) the enteric nervous ganglia.
Tan, J Y; Chua, C K; Leong, K F
2013-02-01
Advanced scaffold fabrication techniques such as Rapid Prototyping (RP) are generally recognized to be advantageous over conventional fabrication methods in terms architectural control and reproducibility. Yet, most RP techniques tend to suffer from resolution limitations which result in scaffolds with uncontrollable, random-size pores and low porosity, albeit having interconnected channels which is characteristically present in most RP scaffolds. With the increasing number of studies demonstrating the profound influences of scaffold pore architecture on cell behavior and overall tissue growth, a scaffold fabrication method with sufficient architectural control becomes imperative. The present study demonstrates the use of RP fabrication techniques to create scaffolds having interconnected channels as well as controllable micro-size pores. Adopted from the concepts of porogen leaching and indirect RP techniques, the proposed fabrication method uses monodisperse microspheres to create an ordered, hexagonal closed packed (HCP) array of micro-pores that surrounds the existing channels of the RP scaffold. The pore structure of the scaffold is shaped using a single sacrificial construct which comprises the microspheres and a dissolvable RP mold that were sintered together. As such, the size of pores as well as the channel configuration of the scaffold can be tailored based on the design of the RP mold and the size of microspheres used. The fabrication method developed in this work can be a promising alternative way of preparing scaffolds with customized pore structures that may be required for specific studies concerning cell-scaffold interactions.
Ratanavaraporn, Juthamas; Kanokpanont, Sorada; Damrongsakkul, Siriporn
2014-02-01
The objective of this study was to develop the microspheres from gelatin (G) and silk fibroin (SF) aimed to be applied for the controlled release of curcumin and piperine. The glutaraldehyde-crosslinked G/SF microspheres at various weight blending ratios (100/0, 70/30, 50/50, and 30/70) were successfully fabricated by water in oil emulsion technique. The microspheres prepared from all compositions were in a round shape with homogeneous size distribution both in the dried (194-217 μm) and swollen states (297-367 μm). When subjected in collagenase solution at physiological condition, the G microspheres gradually degraded within 14 days while the blended G/SF microspheres, particularly at 50/50 and 30/70, were not degraded. For the release application, the microspheres were loaded with curcumin and/or piperine. It was found that the microspheres composed of SF tended to entrap curcumin and piperine with the high entrapment and loading efficiencies, possibly due to their hydrophobic interactions. The G/SF microspheres, particularly at the ratios of 50/50 and 30/70, released curcumin and piperine in a sustained manner both for the single and dual release systems. The controlled dual release of curcumin and piperine from the G/SF microspheres would prolong their half-life, provide the optimal concentrations for therapeutic effects at a target site, and improve the bioavailability of curcumin. These novel injectable microspheres dually releasing curcumin and piperine would be introduced for the treatment of diseases without the need of operation.
Setia, Anupama; Kansal, Sahil; Goyal, Naveen
2013-01-01
Background: Microspheres constitute an important part of oral drug delivery system by virtue of their small size and efficient carrier capacity. However, the success of these microspheres is limited due to their short residence time at the site of absorption. Objective: The objective of the present study was to formulate and systematically evaluate in vitro performance of enteric coated mucoadhesive microspheres of duloxetine hydrochloride (DLX), an acid labile drug. Materials and Methods: DLX microspheres were prepared by simple emulsification phase separation technique using chitosan as carrier and glutaraldehyde as a cross-linking agent. Microspheres prepared were coated with eudragit L-100 using an oil-in-oil solvent evaporation method. Eudragit L-100was used as enteric coating polymer with the aim to release the drug in small intestine The microspheres prepared were characterized by particle size, entrapment efficiency, swelling index (SI), mucoadhesion time, in vitro drug release and surface morphology. A 32 full factorial design was employed to study the effect of independent variables polymer-to-drug ratio (X1) and stirring speed (X2) on dependent variables, particle size, entrapment efficiency, SI, in vitro mucoadhesion and drug release up to 24 h (t24). Results: Microspheres formed were discrete, spherical and free flowing. The microspheres exhibited good mucoadhesive property and also showed high percentage entrapment efficiency. The microspheres were able to sustain the drug release up to 24 h. Conclusion: Thus, the prepared enteric coated mucoadhesive microspheres may prove to be a potential controlled release formulation of DLX for oral administration. PMID:24167786
Efthimiadou, E K; Tziveleka, L-A; Bilalis, P; Kordas, G
2012-05-30
In the current study, poly lactic acid (PLA) modified hollow crosslinked poly(hydroxyethyl methacrylate) (PHEMA) microspheres have been prepared, in order to obtain a stimulus-responsive, biocompatible carrier with sustained drug release properties. The synthetical process consisted of the preparation of poly(methacrylic acid)@poly(hydroxyethyl methacrylate-co-N,N'-methylene bis(acrylamide)) microspheres by a two stage distillation-precipitation polymerization technique using 2,2'-azobisisobutyronitrile as initiator. Following core removal, a PLA coating of the microspheres was formed, after ring opening polymerization of DL-lactide, attributing the initiator's role to the active hydroxyl groups of PHEMA. The anticancer drug daunorubicin (DNR) was selected for the study of loading and release behavior of the coated microspheres. The loading capacity of the PLA modified microspheres was found to be four times higher than that of the parent ones (16% compared to 4%). This coated microspherical carrier exhibited a moderate pH responsive drug release behavior due to the pH dependent water uptake of PHEMA, and PLA hydrolysis. The in vitro cytotoxicity of both the parent and the DNR-loaded or empty modified hollow microspheres has been also examined on MCF-7 breast cancer cells. The results showed that although the empty microspheres were moderately cytotoxic, the DNR-loaded microspheres had more potent anti-tumor effect than the free drug. Therefore, the prepared coated microspheres are interesting drug delivery systems. Copyright © 2012 Elsevier B.V. All rights reserved.
Xiao, Wei; Zhou, Wenjie; Feng, Tong; Zhang, Yanhua; Liu, Hongdong; Tian, Liangliang
2016-09-20
MoS₂/RGO composite hollow microspheres were hydrothermally synthesized by using SiO₂/GO microspheres as a template, which were obtained via the sonication-assisted interfacial self-assembly of tiny GO sheets on positively charged SiO₂ microspheres. The structure, morphology, phase, and chemical composition of MoS₂/RGO hollow microspheres were systematically investigated by a series of techniques such as FE-SEM, TEM, XRD, TGA, BET, and Raman characterizations, meanwhile, their electrochemical properties were carefully evaluated by CV, GCD, and EIS measurements. It was found that MoS₂/RGO hollow microspheres possessed unique porous hollow architecture with high-level hierarchy and large specific surface area up to 63.7 m²·g -1 . When used as supercapacitor electrode material, MoS₂/RGO hollow microspheres delivered a maximum specific capacitance of 218.1 F·g -1 at the current density of 1 A·g -1 , which was much higher than that of contrastive bare MoS₂ microspheres developed in the present work and most of other reported MoS₂-based materials. The enhancement of supercapacitive behaviors of MoS₂/RGO hollow microspheres was likely due to the improved conductivity together with their distinct structure and morphology, which not only promoted the charge transport but also facilitated the electrolyte diffusion. Moreover, MoS₂/RGO hollow microsphere electrode displayed satisfactory long-term stability with 91.8% retention of the initial capacitance after 1000 charge/discharge cycles at the current density of 3 A·g -1 , showing excellent application potential.
In situ fabrication of hollow hydroxyapatite microspheres by phosphate solution immersion
NASA Astrophysics Data System (ADS)
Wang, Yingchun; Yao, Aihua; Huang, Wenhai; Wang, Deping; zhou, Jun
2011-07-01
Hollow hydroxyapatite (HAP) microspheres with pores on their surfaces were prepared by converting Li 2O-CaO-B 2O 3 (LCB) glass microspheres in phosphate solution. The structure, phase composition, surface morphology, and porosity of the hollow HAP microspheres were characterized by SEM, SEM-EDS, XRD, FTIR, ICP-AES, and N 2 adsorption-desorption techniques. The formation and conversion mechanism of the hollow HAP microspheres during immersion process were discussed. The as-prepared microspheres consisted of calcium deficient carbonated hydroxyapatite, which is biomimetic. FTIR spectra indicated that the resulting apatite were B-type CO 3HAP, in which carbonate ions occupied the phosphate sites. After 600 °C heating treatment, hollow microspheres were completely composed of calcium deficient hydroxyapatite crystals including CO32-. The pore size distribution of the as-prepared hollow HAP microspheres were mainly the mesopores in the range of 2-40 nm with the pore volume 0.5614 cm 3/g, and the mean pore size 10.5 nm, respectively. The results confirmed that LCB glass were transformed to hydroxyapatite without changing the external shape and dimension of the original glass object and the resulting microspheres possessed good hollow structures. Once immersed in phosphate solution, Ca-P-OH hydrates were in situ formed on the surface of the glass and precipitated in the position occupied by Ca 2+, while the pores were formed in the position occupied by Li + and B 3+. These hollow HAP microspheres with such structures may be used as promising drug delivery devices.
Sieracki, M E; Reichenbach, S E; Webb, K L
1989-01-01
The accurate measurement of bacterial and protistan cell biomass is necessary for understanding their population and trophic dynamics in nature. Direct measurement of fluorescently stained cells is often the method of choice. The tedium of making such measurements visually on the large numbers of cells required has prompted the use of automatic image analysis for this purpose. Accurate measurements by image analysis require an accurate, reliable method of segmenting the image, that is, distinguishing the brightly fluorescing cells from a dark background. This is commonly done by visually choosing a threshold intensity value which most closely coincides with the outline of the cells as perceived by the operator. Ideally, an automated method based on the cell image characteristics should be used. Since the optical nature of edges in images of light-emitting, microscopic fluorescent objects is different from that of images generated by transmitted or reflected light, it seemed that automatic segmentation of such images may require special considerations. We tested nine automated threshold selection methods using standard fluorescent microspheres ranging in size and fluorescence intensity and fluorochrome-stained samples of cells from cultures of cyanobacteria, flagellates, and ciliates. The methods included several variations based on the maximum intensity gradient of the sphere profile (first derivative), the minimum in the second derivative of the sphere profile, the minimum of the image histogram, and the midpoint intensity. Our results indicated that thresholds determined visually and by first-derivative methods tended to overestimate the threshold, causing an underestimation of microsphere size. The method based on the minimum of the second derivative of the profile yielded the most accurate area estimates for spheres of different sizes and brightnesses and for four of the five cell types tested. A simple model of the optical properties of fluorescing objects and the video acquisition system is described which explains how the second derivative best approximates the position of the edge. Images PMID:2516431
Flexible Microsphere-Embedded Film for Microsphere-Enhanced Raman Spectroscopy.
Xing, Cheng; Yan, Yinzhou; Feng, Chao; Xu, Jiayu; Dong, Peng; Guan, Wei; Zeng, Yong; Zhao, Yan; Jiang, Yijian
2017-09-27
Dielectric microspheres with extraordinary microscale optical properties, such as photonic nanojets, optical whispering-gallery modes (WGMs), and directional antennas, have drawn interest in many research fields. Microsphere-enhanced Raman spectroscopy (MERS) is an alternative approach for enhanced Raman detection by dielectric microstructures. Unfortunately, fabrication of microsphere monolayer arrays is the major challenge of MERS for practical applications on various specimen surfaces. Here we report a microsphere-embedded film (MF) by immersing a highly refractive microsphere monolayer array in the poly(dimethylsiloxane) (PDMS) film as a flexible MERS sensing platform for one- to three-dimensional (1D to 3D) specimen surfaces. The directional antennas and wave-guided whispering-gallery modes (WG-WGMs) contribute to the majority of Raman enhancement by the MFs. Moreover, the MF can be coupled with surface-enhanced Raman spectroscopy (SERS) to provide an extra >10-fold enhancement. The limit of detection is therefore improved for sensing of crystal violet (CV) and Sudan I molecules in aqueous solutions at concentrations down to 10 -7 M. A hybrid dual-layer microsphere enhancer, constructed by depositing a MF onto a microsphere monolayer array, is also demonstrated, wherein the WG-WGMs become dominant and boost the enhancement ratio >50-fold. The present work opens up new opportunities for design of cost-effective and flexible MERS sensing platforms as individual or associated techniques toward practical applications in ultrasensitive Raman detection.
Hu, Meihua; Huang, Pengcheng; Suo, Lili; Wu, Fangying
2017-06-01
A new method based on cetylpyridinium chloride coated ferroferric oxide/silica magnetic microspheres as an efficient solid-phase adsorbent was developed for the extraction and enrichment of ochratoxin A. The determination of ochratoxin A was obtained by high-performance liquid chromatography with fluorescence detection. In the presence of cetylpyridinium chloride, the adsorption capacity of ferroferric oxide/silica microspheres was 5.95 mg/g for ochratoxin A. The experimental parameters were optimized, including the amounts of ferroferric oxide/silica microspheres (20 mg) and cetylpyridinium chloride (0.18 mL, 0.5 mg/mL), pH value of media (9), ultrasonic time (5 min), elution solvent and volume [2(1 + 1) mL (washed twice, 1 mL each time) 1% acetic acid acetonitrile]. Under optimal experiment conditions, ochratoxin A had good linearity in the range of 2.5-250.0 ng/L in water samples with correlation coefficient of the calibration curve 0.9995. The limit of detection for ochratoxin A was 0.83 ng/L, and the recoveries were 89.8-96.8% with the relative standard deviation of 1.5-3.5% in environmental water samples. Furthermore, ferroferric oxide/silica microspheres show excellent reusability during extraction procedures for no less than six times. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Nafissi Varcheh, Nastaran; Aboofazeli, Reza
2011-01-01
The delivery of therapeutic proteins has gained momentum with development of biotechnology. However, large molecular weight, hydrophilic nature and susceptibility to harsh environment of gastrointestinal tract (GIT) resulted in low absorption. The main objective of this work was the design of a particulate system for oral delivery of recombinant human growth hormone (rhGH) on the basis of particle uptake mechanism in GIT. Biodegradable protein-loaded microspheres were prepared using Resomers (RG207, RG756 and RG505) by double emulsion methods. Aqueous solution of protein and freshly prepared rhGH-zinc complex were used for loading process. Various analytical methods, including fluorescence spectroscopy, SDS-PAGE electrophoresis and reversed-phase chromatography, were set up for the quantification and qualification of rhGH before and after the formulation and fabrication procedures. At the optimum conditions, microspheres were mostly below 10 μm with relatively high protein loading (> 50%). Obtained data showed that the stability of protein did not change during the formulation and microencapsulation processes. Results also showed that the encapsulation process in the presence of zinc caused no detectable change in the protein chemical stability. In-vitro stability study of microspheres in different simulated GI media indicated that the entrapped protein was physically stable. Less than 20% of rhGH was released from the microspheres incubated in both simulated stomach and intestine fluids for 3 and 6 h, respectively. PMID:24250342
NASA Astrophysics Data System (ADS)
Harvey, R. W.; Metge, D. W.; Mohanram, A.; Gao, X.; Chorover, J.
2010-12-01
Susceptibilities for in-situ re-entrainment of attached 0.2 and 1.0 μm (diameter) microspheres and groundwater bacteria (Pseudomonas stuzeri and uncultured, native bacteria) were assessed during transport studies involving an organically contaminated, sandy aquifer in Cape Cod, MA. Aquifer sediments between pairs of injection and sampling wells were initially loaded with fluorescently labeled, carboxylated microspheres and bacteria that had been stained with the DNA-specific fluorochrome 4',6-diamidino-2-phenylindole. In response to subsequent hydrodynamic perturbations and injections of deionized water (ionic strength reduction), anionic surfactants (77 μM linear alkylbenzene sulfonates, LAS) and non-ionic surfactant (76 μM polyoxyethylene sorbitan monooleate, Tween 80), differing patterns of re-entrainment were evident for the two colloids. Injections of anionic surfactant and deionized water were the most efficient in causing detachment of the highly hydrophilic and negatively charged microspheres, but largely ineffective in causing re-entrainment of bacteria. In contrast, the nonionic surfactant was highly effective in re-entraining bacteria, but not microspheres. The hydrophobicities and zeta potentials of the indigenous bacteria were highly sensitive to modest concentration changes (0.6 to 1.3 mg L-1) in groundwater dissolved organic carbon (DOC), whereas the microspheres were largely unaffected. The most hydrophilic and negatively charged bacterial community was isolated from groundwater having the lowest DOC. FTIR spectra indicated that the community from the lowest DOC groundwater also had the highest average density of surface carboxyl groups. This indicates that DOC may have a biological effect on native bacteria resulting in changes to surface structures or changes in the makeup of the bacterial community.
Singh, Milind; Morris, Casey P.; Ellis, Ryan J.; Detamore, Michael S.
2008-01-01
Spatial and temporal control of bioactive signals in three-dimensional (3D) tissue engineering scaffolds is greatly desired. Coupled together, these attributes may mimic and maintain complex signal patterns, such as those observed during axonal regeneration or neovascularization. Seamless polymer constructs may provide a route to achieve spatial control of signal distribution. In this study, a novel microparticle-based scaffold fabrication technique is introduced as a method to create 3D scaffolds with spatial control over model dyes using uniform poly(D,L-lactide-co-glycolide) microspheres. Uniform microspheres were produced using the Precision Particle Fabrication technique. Scaffolds were assembled by flowing microsphere suspensions into a cylindrical glass mold, and then microspheres were physically attached to form a continuous scaffold using ethanol treatment. An ethanol soak of 1 h was found to be optimum for improved mechanical characteristics. Morphological and physical characterization of the scaffolds revealed that microsphere matrices were porous (41.1 ± 2.1%) and well connected, and their compressive stiffness ranged from 142 to 306 kPa. Culturing chondrocytes on the scaffolds revealed the compatibility of these substrates with cell attachment and viability. In addition, bilayered, multilayered, and gradient scaffolds were fabricated, exhibiting excellent spatial control and resolution. Such novel scaffolds can serve as sustained delivery devices of heterogeneous signals in a continuous and seamless manner, and may be particularly useful in future interfacial tissue engineering investigations. PMID:18795865
Real-time spectral imaging in three spatial dimensions
NASA Astrophysics Data System (ADS)
Liu, Wenhai; Psaltis, Demetri; Barbastathis, George
2002-05-01
We report what is to our knowledge the first volume-holographic optical imaging instrument with the capability to return three-dimensional spatial as well as spectral information about semitranslucent microscopic objects in a single measurement. The four-dimensional volume-holographic microscope is characterized theoretically and experimentally by use of fluorescent microspheres as objects.
USDA-ARS?s Scientific Manuscript database
Rift Valley Fever virus (RVFV) is a zoonotic virus that infects ruminants including cattle, sheep, goats, camels and buffalo. Multiplexing diagnostic assays that can simultaneously detect antibodies against multiple RVFV antigens offer a high throughput test for disease surveillance and vaccine eva...
NASA Astrophysics Data System (ADS)
Falzone, Tobias; Blair, Savanna; Robertson-Anderson, Rae
2014-03-01
The semi-flexible biopolymer actin is a ubiquitous component of nearly all biological organisms, playing an important role in many biological processes such as cell structure and motility, cancer invasion and metastasis, muscle contraction, and cell signaling. Concentrated actin networks possess unique viscoelastic properties that have been the subject of much theoretical and experimental work. However, much is still unknown regarding the correlation of the applied stress on the network to the induced filament strain at the molecular level. Here, we use dual optical traps alongside fluorescence microscopy to carry out active microrheology measurements that link mechanical stress to structural response at the micron scale. Specifically, we actively drive microspheres through entangled actin networks while simultaneously measuring the force the surrounding filaments exert on the sphere and visualizing the deformation and subsequent relaxation of fluorescent labeled filaments within the network. These measurements, which provide much needed insight into the link between stress and strain in actin networks, are critical for clarifying our theoretical understanding of the complex viscoelastic behavior exhibited in actin networks.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Katalenich, Jeffrey A.; Kitchen, Brian B.; Pierson, Bruce
Cerium dioxide microspheres with uniform diameters between 65 – 211 µm were fabricated using internal gelation sol-gel methods. Although uniform microspheres are produced for nuclear fuel applications with diameters above 300 µm, sol-gel microspheres with diameters of 50 - 200 µm have historically been made by emulsion techniques and had poor size uniformity [1, 2]. An internal gelation, sol-gel apparatus was designed and constructed to accommodate the production of small, uniform microspheres whereby cerium-containing solutions were dispersed into flowing silicone oil and heated in a gelation column to initiate solidification [3, 4]. Problems with premature feed gelation and microsphere coalescencemore » were overcome by equipment modifications unique among known internal gelation setups. Microspheres were fabricated and sized in batches as a function of dispersing needle diameter and silicone oil flow rate in the two-fluid nozzle in order to determine the range of sizes possible and corresponding degree of monodispersity. Initial experiments with poor size uniformity were linked to microsphere coalescence in the gelation column prior to solidification as well as excessive flow rates for the cerium feed solution. Average diameter standard deviations as low as 2.23% were observed after optimization of flow rates and minimization of coalescence reactions.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
Katalenich, Jeffrey A.; Kitchen, Brian B.; Pierson, Bruce D.
Cerium dioxide microspheres with uniform diameters between 65 – 211 µm were fabricated using internal gelation sol-gel methods. Although uniform microspheres are produced for nuclear fuel applications with diameters above 300 µm, sol-gel microspheres with diameters of 50 - 200 µm have historically been made by emulsion techniques and had poor size uniformity [1, 2]. An internal gelation, sol-gel apparatus was designed and constructed to accommodate the production of small, uniform microspheres whereby cerium-containing solutions were dispersed into flowing silicone oil and heated in a gelation column to initiate solidification [3, 4]. Problems with premature feed gelation and microsphere coalescencemore » were overcome by equipment modifications unique among known internal gelation setups. Microspheres were fabricated and sized in batches as a function of dispersing needle diameter and silicone oil flow rate in the two-fluid nozzle in order to determine the range of sizes possible and corresponding degree of monodispersity. Initial experiments with poor size uniformity were linked to microsphere coalescence in the gelation column prior to solidification as well as excessive flow rates for the cerium feed solution. Average diameter standard deviations as low as 2.23% were observed after optimization of flow rates and minimization of coalescence reactions.« less
Magnetic poly(glycidyl methacrylate) microspheres for protein capture.
Koubková, Jana; Müller, Petr; Hlídková, Helena; Plichta, Zdeněk; Proks, Vladimír; Vojtěšek, Bořivoj; Horák, Daniel
2014-09-25
The efficient isolation and concentration of protein antigens from complex biological samples is a critical step in several analytical methods, such as mass spectrometry, flow cytometry and immunochemistry. These techniques take advantage of magnetic microspheres as immunosorbents. The focus of this study was on the development of new superparamagnetic polymer microspheres for the specific isolation of the tumor suppressor protein p53. Monodisperse macroporous poly(glycidyl methacrylate) (PGMA) microspheres measuring approximately 5 μm and containing carboxyl groups were prepared by multistep swelling polymerization of glycidyl methacrylate (GMA), 2-[(methoxycarbonyl)methoxy]ethyl methacrylate (MCMEMA) and ethylene dimethylacrylate (EDMA) as a crosslinker in the presence of cyclohexyl acetate as a porogen. To render the microspheres magnetic, iron oxide was precipitated within their pores; the Fe content in the particles received ∼18 wt%. Nonspecific interactions between the magnetic particles and biological media were minimized by coating the microspheres with poly(ethylene glycol) (PEG) terminated by carboxyl groups. The carboxyl groups of the magnetic PGMA microspheres were conjugated with primary amino groups of mouse monoclonal DO-1 antibody using conventional carbodiimide chemistry. The efficiency of protein p53 capture and the degree of nonspecific adsorption on neat and PEG-coated magnetic microspheres were determined by western blot analysis. Copyright © 2014 Elsevier B.V. All rights reserved.
Composite nuclear fuel fabrication methodology for gas fast reactors
NASA Astrophysics Data System (ADS)
Vasudevamurthy, Gokul
An advanced fuel form for use in Gas Fast Reactors (GFR) was investigated. Criteria for the fuel includes operation at high temperature (˜1400°C) and high burnup (˜150 MWD/MTHM) with effective retention of fission products even during transient temperatures exceeding 1600°C. The GFR fuel is expected to contain up to 20% transuranics for a closed fuel cycle. Earlier evaluations of reference fuels for the GFR have included ceramic-ceramic (cercer) dispersion type composite fuels of mixed carbide or nitride microspheres coated with SiC in a SiC matrix. Studies have indicated that ZrC is a potential replacement for SiC on account of its higher melting point, increased fission product corrosion resistance and better chemical stability. The present work investigated natural uranium carbide microspheres in a ZrC matrix instead of SiC. Known issues of minor actinide volatility during traditional fabrication procedures necessitated the investigation of still high temperature but more rapid fabrication techniques to minimize these anticipated losses. In this regard, fabrication of ZrC matrix by combustion synthesis from zirconium and graphite powders was studied. Criteria were established to obtain sufficient matrix density with UC microsphere volume fractions up to 30%. Tests involving production of microspheres by spark erosion method (similar to electrodischarge machining) showed the inability of the method to produce UC microspheres in the desired range of 300 to 1200 mum. A rotating electrode device was developed using a minimum current of 80A and rotating at speeds up to 1500 rpm to fabricate microspheres between 355 and 1200 mum. Using the ZrC process knowledge, UC electrodes were fabricated and studied for use in the rotating electrode device to produce UC microspheres. Fabrication of the cercer composite form was studied using microsphere volume fractions of 10%, 20%, and 30%. The macrostructure of the composite and individual components at various stages were characterized to understand the required fabrication techniques and at the same time meet the necessary GFR fuel characteristics.
Whispering gallery modes in two-photon fluorescence from spherical DCM dye microresonators
NASA Astrophysics Data System (ADS)
Mamonov, Evgeniy A.; Maydykovskiy, Anton I.; Mitetelo, Nikolai V.; Venkatakrishnarao, Dasari; Chandrasekar, Rajadurai; Murzina, Tatyana V.
2018-03-01
Organic microstructures are well known for their resonator properties, which bring about whispering gallery mode (WGM) excitation. Here we report on experimental evidence of the WGM in the two-photon fluorescence (TPF) of DCM dye microspheres made using the self-assembly method. The WGM excitation accompanying the overall TPF in the spectral range from 530\\div640 nm demonstrated a quality factor of approximately 102 for spheres that were several microns in diameter. The power dependence of the TPF intensity proved the second order nature of the interaction process involved.
Hammond, G. Denise; Vojta, Adam L.; Grant, Sheila A.; Hunt, Heather K.
2016-01-01
The creation of label-free biosensors capable of accurately detecting trace contaminants, particularly small organic molecules, is of significant interest for applications in environmental monitoring. This is achieved by pairing a high-sensitivity signal transducer with a biorecognition element that imparts selectivity towards the compound of interest. However, many environmental pollutants do not have corresponding biorecognition elements. Fortunately, biomimetic chemistries, such as molecular imprinting, allow for the design of artificial receptors with very high selectivity for the target. Here, we perform a proof-of-concept study to show how artificial receptors may be created from inorganic silanes using the molecular imprinting technique and paired with high-sensitivity transducers without loss of device performance. Silica microsphere Whispering Gallery Mode optical microresonators are coated with a silica thin film templated by a small fluorescent dye, fluorescein isothiocyanate, which serves as our model target. Oxygen plasma degradation and solvent extraction of the template are compared. Extracted optical devices are interacted with the template molecule to confirm successful sorption of the template. Surface characterization is accomplished via fluorescence and optical microscopy, ellipsometry, optical profilometry, and contact angle measurements. The quality factors of the devices are measured to evaluate the impact of the coating on device sensitivity. The resulting devices show uniform surface coating with no microstructural damage with Q factors above 106. This is the first report demonstrating the integration of these devices with molecular imprinting techniques, and could lead to new routes to biosensor creation for environmental monitoring. PMID:27314397
Elbert, Donald L.
2010-01-01
Macroporous hydrogels may have direct applications in regenerative medicine as scaffolds to support tissue formation. Hydrogel microspheres may be used as drug delivery vehicles or as building blocks to assemble modular scaffolds. A variety of techniques exist to produce macroporous hydrogels and hydrogel microspheres. A subset of these relies on liquid-liquid two phase systems. Within this subset, vastly different types of polymerization processes are found. In this review, the history, terminology and classification of liquid-liquid two phase polymerization and crosslinking are described. Instructive examples of hydrogel microsphere and macroporous scaffold formation by precipitation/dispersion, emulsion and suspension polymerizations are used to illustrate the nature of these processes. The role of the kinetics of phase separation in determining the morphology of scaffolds and microspheres is also delineated. Brief descriptions of miniemulsion, microemulsion polymerization and ionotropic gelation are also included. PMID:20659596
[Synthesis of hollow titania microspheres by using microfluidic droplet-template].
Ma, Jingyun; Jiang, Lei; Qin, Jianhu
2011-09-01
Droplet-based microfluidics is of great interest due to its particular characteristics compared with the conventional methods, such as reduced reagent consumption, rapid mixing, high-throughput, shape controlled, etc. A novel method using microfluidic droplet as soft template for the synthesis of hollow titania microspheres was developed. A typical polydimethylsiloxane (PDMS) microfluidic device containing "flow-focusing" geometry was used to generate water/oil (W/O) droplet. The mechanism for the hollow structure formation was based on the interfacial hydrolysis reaction between the continuous phase containing titanium butoxide precursor and the dispersed containing water. The continuous phase mixed with butanol was added in the downstream of the channel after the hydrolysis reaction. This step was used for drawing the water out of the microgels for further hydrolysis. The microgels obtained through a glass pipe integrated were washed, dried under vacuum and calcined after aging for a certain time. The fluorescence and scanning electron microscope (SEM) image of the microspheres indicated the hollow structure and the thickness of the shell. In addition, these microspheres with thin shell (about 2 microm) were apt to rupture and collapse. Droplet-based microfluidic offered a gentle and size-controllable manner to moderate this problem. Moreover, it has potential applications in photocatalysis combined with some modification realized on the chip simultaneously.
Hosseini, Samira; Aeinehvand, Mohammad M; Uddin, Shah M; Benzina, Abderazak; Rothan, Hussin A; Yusof, Rohana; Koole, Leo H; Madou, Marc J; Djordjevic, Ivan; Ibrahim, Fatimah
2015-11-09
The application of microfluidic devices in diagnostic systems is well-established in contemporary research. Large specific surface area of microspheres, on the other hand, has secured an important position for their use in bioanalytical assays. Herein, we report a combination of microspheres and microfluidic disk in a unique hybrid platform for highly sensitive and selective detection of dengue virus. Surface engineered polymethacrylate microspheres with carefully designed functional groups facilitate biorecognition in a multitude manner. In order to maximize the utility of the microspheres' specific surface area in biomolecular interaction, the microfluidic disk was equipped with a micromixing system. The mixing mechanism (microballoon mixing) enhances the number of molecular encounters between spheres and target analyte by accessing the entire sample volume more effectively, which subsequently results in signal amplification. Significant reduction of incubation time along with considerable lower detection limits were the prime motivations for the integration of microspheres inside the microfluidic disk. Lengthy incubations of routine analytical assays were reduced from 2 hours to 5 minutes while developed system successfully detected a few units of dengue virus. Obtained results make this hybrid microsphere-microfluidic approach to dengue detection a promising avenue for early detection of this fatal illness.
NASA Astrophysics Data System (ADS)
Abdel-Fattah, A. N.; Langford, R.; Schulze-Makuch, D.; Sheng, Z.
2005-12-01
River bank filtered water is an important component of the drinking water production in many areas of the world. In riverbank filtration, the removal of pathogens is an important task for the production of good quality drinking water. The hydrogeological factors and spatial changes in the water's microbiology during the transport from the river to the aquifer have important implications on the quality of the produced water. The goal of this study was to investigate riverbank infiltration effectiveness in arid environments such as that of El Paso, Texas. The hydrostratigraphic units and hydrogeologic conditions were characterized with lithologic samples obtained from all boreholes collected during the construction of twelve observation wells and one production well in the site, which were constructed near the artificial stream to provide geologic and hydrologic information. The shallow aquifer is composed of three unites: high hydraulic conductivity layers on the top and bottom, and low conductivity layer in the middle. In this study advective transport of microspheres was compared with a conservative tracer such as bromide. Bromide was injected into an observation well at the channel margin. Simultaneously, 1, 6 and 10 micron-diameter fluorescent microspheres equivalent to Giardia, Cryptosporidium, and bacteria sizes were injected into the stream bottom and two observation wells to assess the suitability of microspheres as abiotic analogs in future investigations involving the physical aspects of bacteria and protozoa transport behavior. The 17.8 day-tracer test provided valuable results that are relevant to the transport of pathogens through the subsurface under riverbank filtration conditions. The 1 micron-size microspheres were abundant in the pumping and observation wells and showed multiple peaks similar to the bromide results. Microspheres from the three injection sites had distinctly different transport paths and rates. The 6 and 10 micron-size microspheres were observed in low to almost zero concentrations. The very low detection of larger sized microspheres indicates a potential attenuation of Cryptosporidium and Giardia-size microspheres under riverbank filtration conditions.
Mochalov, Konstantin E; Chistyakov, Anton A; Solovyeva, Daria O; Mezin, Alexey V; Oleinikov, Vladimir A; Vaskan, Ivan S; Molinari, Michael; Agapov, Igor I; Nabiev, Igor; Efimov, Anton E
2017-11-01
In the past decade correlative microscopy, which combines the potentials of different types of high-resolution microscopies with a variety of optical microspectroscopy techniques, has been attracting increasing attention in material science and biological research. One of outstanding solutions in this area is the combination of scanning probe microscopy (SPM), which provides data on not only the topography, but also the spatial distribution of a wide range of physical properties (elasticity, conductivity, etc.), with ultramicrotomy, allowing 3D multiparametric examination of materials. The combination of SPM and ultramicrotomy (scanning probe nanotomography) is very appropriate for characterization of soft multicompound nanostructurized materials, such as polymer matrices and microstructures doped with different types of nanoparticles (magnetic nanoparticles, quantum dots, nanotubes, etc.), and biological materials. A serious problem of this technique is a lack of chemical and optical characterization tools, which may be solved by using optical microspectroscopy. Here, we report the development of an instrumental approach to combining confocal microspectroscopy and 3D scanning probe nanotomography in a single apparatus. This approach retains all the advantages of SPM and upright optical microspectroscopy and allows 3D multiparametric characterization using both techniques. As the first test of the system developed, we have performed correlative characterization of the morphology and the magnetic and fluorescent properties of fluorescent magnetic microspheres doped with a fluorescent dye and magnetic nanoparticles. The results of this study can be used to obtain 3D volume images of a specimen for most high-resolution near-field scanning probe microscopies: SNOM, TERS, AFM-IR, etc. This approach will result in development of unique techniques combining the advantages of SPM (nanoscale morphology and a wide range of physical parameters) and high-resolution optical microspectroscopy (nanoscale chemical mapping and optical properties) and allowing simultaneous 3D measurements. Copyright © 2017 Elsevier B.V. All rights reserved.
Epi-illumination optical design for fluorescence polarization measurements in flow systems.
Eisert, W G; Beisker, W
1980-01-01
An epi-illumination design for fluorescence polarization measurements is introduced in flow cytometry with the optical axis orthogonally aligned to the cell stream. Various optical components and designs are discussed with respect to their influence on polarization measurements. Using the epi-configuration, paired measurements with the direction of polarization of the exciting light changed orthogonally are proposed for the compensation of system anisotropies and electronic mismatch. Large aperture corrections are employed for the excitation as well as for the emission pathway. Additional parameters such as fluorescence at 90 degrees, multiangle light scattering, and high precision cell-sizing by internally calibrated time of the flight measurements, as described previously, remain available with the design proposed here. Fluorescent latex microspheres, stained intracellular DNA, and algae have been used to test performance. PMID:7023562
Two-dimensional microsphere quasi-crystal: fabrication and properties
NASA Astrophysics Data System (ADS)
Noginova, Natalia E.; Venkateswarlu, Putcha; Kukhtarev, Nickolai V.; Sarkisov, Sergey S.; Noginov, Mikhail A.; Caulfield, H. John; Curley, Michael J.
1996-11-01
2D quasi-crystals were fabricated from polystyrene microspheres and characterized for their structural, diffraction, and non-linear optics properties. The quasi- crystals were produced with the method based on Langmuir- Blodgett thin film technique. Illuminating the crystal with the laser beam, we observed the diffraction pattern in the direction of the beam propagation and in the direction of the back scattering, similar to the x-ray Laue pattern observed in regular crystals with hexagonal structure. The absorption spectrum of the quasi-crystal demonstrated two series of regular maxima and minima, with the spacing inversely proportional to the microspheres diameter. Illumination of the dye-doped microspheres crystal with Q- switched radiation of Nd:YAG laser showed the enhancement of non-linear properties, in particular, second harmonic generation.
Dhaliwal, Sumeet; Jain, Subheet; Singh, Hardevinder P; Tiwary, A K
2008-06-01
The aim of the present investigation was to evaluate the potential use of mucoadhesive microspheres for gastroretentive delivery of acyclovir. Chitosan, thiolated chitosan, Carbopol 71G and Methocel K15M were used as mucoadhesive polymers. Microsphere formulations were prepared using emulsion-chemical crosslinking technique and evaluated in vitro, ex-vivo and in-vivo. Gelatin capsules containing drug powder showed complete dissolution (90.5 +/- 3.6%) in 1 h. The release of drug was prolonged to 12 h (78.8 +/- 3.9) when incorporated into mucoadhesive microspheres. The poor bioavailability of acyclovir is attributed to short retention of its dosage form at the absorption sites (in upper gastrointestinal tract to duodenum and jejunum). The results of mucoadhesion study showed better retention of thiolated chitosan microspheres (8.0 +/- 0.8 h) in duodenal and jejunum regions of intestine. The results of qualitative and quantitative GI distribution study also showed significant higher retention of mucoadhesive microspheres in upper GI tract. Pharmacokinetic study revealed that administration of mucoadhesive microspheres could maintain measurable plasma concentration of acyclovir through 24 h, as compared to 5 h after its administration in solution form. Thiolated chitosan microsphere showed superiority over the other formulations as observed with nearly 4.0-fold higher AUC(0-24) value (1,090 +/- 51 ng h/ml) in comparison to drug solution (281 +/- 28 ng h/ml). Overall, the result indicated prolonged delivery with significant improvement in oral bioavailability of acyclovir from mucoadhesive microspheres due to enhanced retention in the upper GI tract.
Hydrophilic microspheres from water-in-oil emulsions by the water diffusion technique.
Trotta, Michele; Chirio, Daniela; Cavalli, Roberta; Peira, Elena
2004-08-01
In this study, we developed and evaluated a novel method to produce insulin-loaded hydrophilic microspheres allowing high encapsulation efficiency and the preservation of peptide stability during particle processing. The preparation method used the diffusion of water by an excess of solvent starting from a water-in-solvent emulsion. The water dispersed phase containing albumin or lactose, or albumin-lactose in different weight ratios, and insulin was emulsified in water-saturated triacetin with and without emulsifiers, producing a water-in-triacetin emulsion. An excess of triacetin was added to the emulsion so that water could be extracted into the continuous phase, allowing the insulin-loaded microsphere precipitation. Insulin stability within the microspheres after processing was evaluated by reverse-phase and size-exclusion high-performance liquid chromatography. The water diffusion extraction process provided spherical microparticles of albumin or albumin-lactose. The mean diameter of the microspheres prepared with or without emulsifiers ranged from 2 to 10 microm, and the encapsulation efficiency of insulin was between 60% and 75%, respectively. The analysis of microsphere content after processing showed that insulin did not undergo any chemical modification within microspheres. The use of lactose alone led to the formation of highly viscous droplets that coalesced during the purification step. The water extraction procedures successfully produced insulin-loaded hydrophilic microspheres allowing the preservation of peptide stability. The type of excipient and the size of the disperse phase of the primary w/o emulsion were crucial determinants of microsphere characteristics.
Ha, Dong-Ho; Pathak, Shiva; Yong, Chul Soon; Kim, Jong Oh; Jeong, Jee-Heon; Park, Jun-Beom
2016-01-01
The aim of the present study is to evaluate the potential differentiation ability of gingiva originated human mesenchymal stem cell in the presence of tacrolimus. Tacrolimus-loaded poly(lactic-co-glycolic acid) microspheres were prepared using electrospraying technique. In vitro release study of tacrolimus-loaded poly(lactic-co-glycolic acid) microspheres was performed in phosphate-buffered saline (pH 7.4). Gingiva-derived stem cells were isolated and incubated with tacrolimus or tacrolimus-loaded microspheres. Release study of the microspheres revealed prolonged release profiles of tacrolimus without any significant initial burst release. The microsphere itself did not affect the morphology of the mesenchymal stem cells, and cell morphology was retained after incubation with microspheres loaded with tacrolimus at 1 μg/mL to 10 μg/mL. Cultures grown in the presence of microspheres loaded with tacrolimus at 1 μg/mL showed the highest mineralization. Alkaline phosphatase activity increased with an increase in incubation time. The highest expression of pSmad1/5 was achieved in the group receiving tacrolimus 0.1 μg/mL every third day, and the highest expression of osteocalcin was achieved in the group receiving 1 μg/mL every third day. Biodegradable poly(lactic-co-glycolic acid)-based microspheres loaded with tacrolimus promoted mineralization. Microspheres loaded with tacrolimus may be applied for increased osteoblastic differentiation. PMID:27721434
Role of coherence in microsphere-assisted nanoscopy
NASA Astrophysics Data System (ADS)
Perrin, Stephane; Lecler, Sylvain; Leong-Hoi, Audrey; Montgomery, Paul C.
2017-06-01
The loss of the information, due to the diffraction and the evanescent waves, limits the resolving power of classical optical microscopy. In air, the lateral resolution of an optical microscope can approximated at half of the wavelength using a low-coherence illumination. Recently, several methods have been developed in order to overcome this limitation and, in 2011, a new far-field and full-field imaging technique was proposed where a sub-diffraction-limit resolution has been achieved using a transparent microsphere. In this article, the phenomenon of super-resolution using microsphere-assisted microscopy is analysed through rigorous electro-magnetic simulations. The performances of the imaging technique are estimated as function of optical and geometrical parameters. Furthermore, the role of coherence is introduced through the temporal coherence of the light source and the phase response of the object.
Liu, Yuan; Yuan, Baohong; Vignola, Joseph
2012-01-01
To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes.
Liu, Yuan; Yuan, Baohong; Vignola, Joseph
2013-01-01
To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes. PMID:24179476
Epifluorescence Intravital Microscopy of Murine Corneal Dendritic Cells
Rosenbaum, James T.; Planck, Stephen R.
2010-01-01
Purpose. Dendritic cells (DCs) are antigen-presenting cells vital for initiating immune responses. In this study the authors examined the in vivo migratory capability of resident corneal DCs to various stimuli. Methods. The authors used mice expressing enhanced yellow fluorescent protein (eYFP) under control of the CD11c promoter to visualize corneal DCs. To assess the distribution and mobility of DCs, normal corneas were imaged in vivo and ex vivo with fluorescence microscopy. Intravital microscopy was used to examine the responses of resident central and peripheral corneal DCs to silver nitrate injury, lipopolysaccharide, microspheres, and tumor necrosis factor (TNF-α). In some experiments, TNF-α injection was used to first induce centripetal migration of DCs to the central cornea, which was subsequently reinjected with microspheres. Results. In normal corneas, DCs were sparsely distributed centrally and were denser in the periphery, with epithelial-level DCs extending into the epithelium. Videomicroscopy showed that though cell processes were in continuous movement, cells generally did not migrate. Within the first 6 hours after stimulation, neither central nor peripheral corneal DCs exhibited significant lateral migration, but central corneal DCs assumed extreme morphologic changes. An increased number of DCs in the TNF-α–stimulated central cornea were responsive to subsequent microsphere injection by adopting a migratory behavior, but not with increased speed. Conclusions. In vivo imaging reveals minimal lateral migration of corneal DCs after various stimuli. In contrast, DCs within the central cornea after initial TNF-α injection are more likely to respond to a secondary insult with lateral migration. PMID:20007837
Efthimiadou, Eleni K; Tapeinos, Christos; Chatzipavlidis, Alexandros; Boukos, Nikos; Fragogeorgi, Eirini; Palamaris, Lazaros; Loudos, George; Kordas, George
2014-01-30
This paper deals with the synthesis, characterization and property evaluation of drug-loaded magnetic microspheres with pH-responsive cross-linked polymer shell. The synthetic procedure consists of 3 steps, of which the first two comprise the synthesis of a poly methyl methacrylate (PMMA) template and the synthesis of a shell by using acrylic acid (AA) and methyl methacrylate (MMA) as monomers, and divinyl benzene (DVB) as cross-linker. The third step of the procedure refers to the formation of magnetic nanoparticles on the microsphere's surface. AA that attaches pH-sensitivity in the microspheres and magnetic nanoparticles in the inner and the outer surface of the microspheres, enhance the efficacy of this intelligent drug delivery system (DDS), which constitutes a promising approach toward cancer therapy. A number of experimental techniques were used to characterize the resulting microspheres. In order to investigate the in vitro controlled release behavior of the synthesized microspheres, we studied the Dox release percentage under different pH conditions and under external magnetic field. Hyperthermia caused by an alternating magnetic field (AFM) is used in order to study the doxorubicin (Dox) release behavior from microspheres with pH functionality. The in vivo fate of these hybrid-microspheres was tracked by labeling them with the γ-emitting radioisotope (99m)Tc after being intravenously injected in normal mice. According to our results, microsphere present a pH depending and a magnetic heating, release behavior. As expected, labeled microspheres were mainly found in the mononuclear phagocyte system (MPS). The highlights of the current research are: (i) to illustrate the advantages of controlled release by combining hyperthermia and pH-sensitivity and (ii) to provide noninvasive, in vivo information on the spatiotemporal biodistribution of these microsphere by dynamic γ-imaging. Copyright © 2013 Elsevier B.V. All rights reserved.
Hollow silica microspheres for buoyancy-assisted separation of infectious pathogens from stool.
Weigum, Shannon E; Xiang, Lichen; Osta, Erica; Li, Linying; López, Gabriel P
2016-09-30
Separation of cells and microorganisms from complex biological mixtures is a critical first step in many analytical applications ranging from clinical diagnostics to environmental monitoring for food and waterborne contaminants. Yet, existing techniques for cell separation are plagued by high reagent and/or instrumentation costs that limit their use in many remote or resource-poor settings, such as field clinics or developing countries. We developed an innovative approach to isolate infectious pathogens from biological fluids using buoyant hollow silica microspheres that function as "molecular buoys" for affinity-based target capture and separation by floatation. In this process, antibody functionalized glass microspheres are mixed with a complex biological sample, such as stool. When mixing is stopped, the target-bound, low-density microspheres float to the air/liquid surface, which simultaneously isolates and concentrates the target analytes from the sample matrix. The microspheres are highly tunable in terms of size, density, and surface functionality for targeting diverse analytes with separation times of ≤2min in viscous solutions. We have applied the molecular buoy technique for isolation of a protozoan parasite that causes diarrheal illness, Cryptosporidium, directly from stool with separation efficiencies over 90% and low non-specific binding. This low-cost method for phenotypic cell/pathogen separation from complex mixtures is expected to have widespread use in clinical diagnostics as well as basic research. Copyright © 2016 Elsevier B.V. All rights reserved.
Singh, Milind; Dormer, Nathan; Salash, Jean R.; Christian, Jordan M.; Moore, David S.; Berkland, Cory; Detamore, Michael S.
2010-01-01
A novel approach has been demonstrated to construct biocompatible, macroporous 3-D tissue engineering scaffolds containing a continuous macroscopic gradient in composition that yields a stiffness gradient along the axis of the scaffold. Polymeric microspheres, made of poly(d,l-lactic-co-glycolic acid) (PLGA), and composite microspheres encapsulating a higher stiffness nano-phase material (PLGA encapsulating CaCO3 or TiO2 nanoparticles) were used for the construction of microsphere-based scaffolds. Using controlled infusion of polymeric and composite microspheres, gradient scaffolds displaying an anisotropic macroscopic distribution of CaCO3/TiO2 were fabricated via an ethanol sintering technique. The controllable mechanical characteristics and biocompatible nature of these scaffolds warrants further investigation for interfacial tissue engineering applications. PMID:20336753
Du, Pengcheng; Zeng, Jin; Mu, Bin; Liu, Peng
2013-05-06
Well-defined biocompatible magnetic and molecular dual-targeting polyelectrolyte hybrid hollow microspheres have been accomplished via the layer-by-layer (LbL) self-assembly technique. The hybrid shell was fabricated by the electrostatic interaction between the polyelectrolyte cation, chitosan (CS), and the hybrid anion, citrate modified ferroferric oxide nanoparticles (Fe3O4-CA), onto the uniform polystyrene sulfonate microsphere templates. Then the magnetic hybrid core/shell composite particles were modified with a linear, functional poly(ethylene glycol) (PEG) monoterminated with a biotargeting molecule (folic acid (FA)). Afterward the dual targeting hybrid hollow microspheres were obtained after etching the templates by dialysis. The dual targeting hybrid hollow microspheres exhibit exciting pH response and stability in high salt-concentration media. Their pH-dependent controlled release of the drug molecule (anticancer drug, doxorubicin (DOX)) was also investigated in different human body fluids. As expected, the cell viability of the HepG2 cells which decreased more rapidly was treated by the FA modified hybrid hollow microspheres rather than the unmodified one in the in vitro study. The dual-targeting hybrid hollow microspheres demonstrate selective killing of the tumor cells. The precise magnetic and molecular targeting properties and pH-dependent controlled release offers promise for cancer treatment.
Wang, Xuchun; Li, Guangyong; Hong, Guo; Guo, Qiang; Zhang, Xuetong
2017-11-29
Phase change materials, changing from solid to liquid and vice versa, are capable of storing and releasing a large amount of thermal energy during the phase change, and thus hold promise for numerous applications including thermal protection of electronic devices. Shaping these materials into microspheres for additional fascinating properties is efficient but challenging. In this regard, a novel phase change microsphere with the design for electrical-regulation and thermal storage/release properties was fabricated via the combination of monodispersed graphene aerogel microsphere (GAM) and phase change paraffin. A programmable method, i.e., coupling ink jetting-liquid marbling-supercritical drying (ILS) techniques, was demonstrated to produce monodispersed graphene aerogel microspheres (GAMs) with precise size-control. The resulting GAMs showed ultralow density, low electrical resistance, and high specific surface area with only ca. 5% diameter variation coefficient, and exhibited promising performance in smart switches. The phase change microspheres were obtained by capillary filling of phase change paraffin inside the GAMs and exhibited excellent properties, such as low electrical resistance, high latent heat, well sphericity, and thermal buffering. Assembling the phase change microsphere into the microcircuit, we found that this tiny device was quite sensitive and could respond to heat as low as 0.027 J.
Aramwit, Pornanong; Yamdech, Rungnapha; Ampawong, Sumate
2016-05-01
One approach in wound dressing development is to incorporate active molecules or drugs in the dressing. In order to reduce the frequency of dressing changes as well as to prolong wound healing efficacy, wound dressings that can sustain the release of the active molecules should be developed. In our previous work, we developed chitosan/sericin (CH/SS) microspheres that released sericin in a controlled rate. However, the difficulty of applying the microspheres that easily diffuse and quickly degrade onto the wound was its limitations. In this study, we aimed to develop wound dressing materials which are easier to apply and to provide extended release of sericin. Different amounts of CH/SS microspheres were embedded into various compositions of polyvinyl alcohol/gelatin (PVA/G) scaffolds and fabricated using freeze-drying and glutaraldehyde crosslinking techniques. The obtained CH/SS microspheres-embedded scaffolds with appropriate design and formulation were introduced as a wound dressing material. Sericin was released from the microspheres and the scaffolds in a sustained manner. Furthermore, an optimized formation of the microspheres-embedded scaffolds (2PVA2G+2CHSS) was shown to possess an effective antimicrobial activity against both gram-positive and gram-negative bacteria. These microspheres-embedded scaffolds were not toxic to L929 mouse fibroblast cells, and they did not irritate the tissue when applied to the wound. Finally, probably by the sustained release of sericin, these microspheres-embedded scaffolds could promote wound healing as well as or slightly better than a clinically used wound dressing (Allevyn®) in a mouse model. The antimicrobial CH/SS microspheres-embedded PVA/G scaffolds with sustained release of sericin would appear to be a promising candidate for wound dressing application.
Gavini, E; Alamanni, M C; Cossu, M; Giunchedi, P
2005-08-01
Controlled release dosage forms based on tabletted microspheres containing fresh artichoke Cynara scolymus extract were performed for the oral administration of a nutritional supplement. Microspheres were prepared using a spray-drying technique; lactose or hypromellose have been chosen as excipients. Microspheres were characterized in terms of encapsulated extract content, size and morphology. Qualitative and quantitative composition of the extract before and after the spray process was determined. Compressed matrices (tablets) were prepared by direct compression of the spray-dried microspheres. In vitro release tests of microparticles and tablets prepared were carried out in both acidic and neutral media. Spray-drying is a good method to prepare microspheres containing the artichoke extract. The microspheres encapsulate an amount of extract close to the theoretical value. Particle size analyses indicate that the microparticles have dvs of approximately 6-7 microm. Electronic microscopy observations reveal that particles based on lactose have spherical shape and particles containing hypromellose are almost collapsed. The hydroalcoholic extract is stable to the microsphere production process: its polyphenolic composition (qualitative and quantitative) did not change after spraying. In vitro release studies show that microparticles characterized by a quick polyphenolic release both in acidic and neutral media due to the high water solubility of the carrier lactose. On the contrary, microspheres based hypromellose release only 20% of the loaded extract at pH 1.2 in 2 h and the total amount of polyphenols is released only after about further 6 h at pH 6.8. Matrices prepared tabletting lactose microspheres and hypromellose microparticles in the weight ratio 1:1 show a slow release rate, that lasts approximately 24 h. This one-a-day sustained release formulation containing Cynara scolymus extract could be proposed as a nutraceutical controlled release dosage form for oral administration.
Tian, Jiyuan; Yu, Juan; Sun, Xiuqin
2008-12-15
Oral DNA-based immunotherapy is a new treatment option for fish immunisation in intensive culture. However, because of the existence of the nucleases and severe gastrointestinal conditions, DNA-based vaccines can be hydrolyzed or denatured. In our laboratory, a plasmid DNA (pDNA) containing major capsid protein (MCP) gene of lymphocystis disease virus (LCDV) was prepared, and then pDNA was encapsulated in chitosan microspheres through an emulsion-based methodology. The yield, loading percent and encapsulation efficiency of microspheres were 93.6%, 0.3% and 94.5%, respectively. Scanning electron microscopy (SEM) showed that pDNA-loaded microspheres yielded a spherical shape with smooth surfaces. The disproportion of super-coiled to open circle and linear pDNA suggested that high transfection efficiencies of pDNA in microspheres were retained. The cumulative release of pDNA showed that chitosan microspheres were resistant to degradation in simulated gastrointestinal tract environment. The release profile at PBS buffer (pH 7.4) displayed that pDNA-loaded chitosan microspheres had a release up to 42 days after intestinal imbibition. RT-PCR showed that RNA containing information of MCP gene existed in various tissues 10-90 days post-vaccination. SDS-PAGE and immunofluorescent images indicated that pDNA expressed MCP in tissues of fish 10-90 days after oral administration. In addition, indirect ELISA displayed that the immune responses of sera were positive (O.D.> or =0.3) from week 1 to week 16 for fish vaccinated with microspheres, in comparison with fish vaccinated with naked pDNA. Data obtained suggested that chitosan microspheres were promising carriers for oral pDNA vaccine. Because this encapsulation technique was easy to operate and immunisation efficacy of microspheres loaded with pDNA was significant, it had potential to be used in drug delivery applications.
SU-E-T-402: Y-90 Microspheres (SIR Spheres) for Treatment of Liver Metastasis : Technique
DOE Office of Scientific and Technical Information (OSTI.GOV)
Nair, M
2014-06-01
Purpose: The purpose of this presentation is to discuss the radiation safety and dosimetric technique used for the therapeutic procedure using Y-90 microspheres through intra -arterial administration on patients with liver metastasis Methods: The radiation dosimetry, technique and safety aspects of 14 patients with primary and metastatic liver cancer, treated with Y-90 microsphere (SIR spheres) are discussed. The liver and tumor volumes were determined using the CT and MR scans . The images were imported into the treatment planning system and the liver and tumor volumes and the volume of the liver affected were outlined and the volume calculation wasmore » performed using the software. The lung shunt fraction (LSF) and tumor to liver uptake ratio (TLR) were determined using the nuclear medicine SPECT imaging with Tc-99m MAA. The absorbed dose to the target volume in liver was calculated using the following equation:Dose ? (Gy) = C x E? x 5.92 x 10-6 (Gy/s) x T(1/2)(days) x 1.44 x 8.64 x 104 (s) The distribution of activity in the tumor bed was confirmed by post Y-90 administration imaging using the Bremsstrahlung peak at 30% window. The patient and the procedure room were surveyed and radiation safety instructions were given to the patient Results: The tumor volume ranged from 77 cc to 700 cc, tumor to liver uptake ranged from 3 to 12. The lung shunt fraction varied from 1.08% to 9.0%. The activity administered ranged from 1.0GBq to 2.5 GBq, . The radiation survey in contact with the patient ranged from 1.8 mR/hr to 2.5 mR/hr and reading at 1 meter was less than 0.2 mR/hr Conclusion: The technique for radiation dosimetry and radiation safety for Y-90 microsphere therapy is established. The post treatment imaging helped to confirm the distribution of Y-90 microspheres inside the tumor bed.« less
Development and evaluation of intestinal targeted mucoadhesive microspheres of Bacillus coagulans.
Alli, Sk Md Athar; Ali, Sk Md Ajhar; Samanta, Amalesh
2011-11-01
Intestinal targeted mucoadhesive microsphere of probiotics may provide numerous associated health benefits. To develop mucoadhesive microspheres that will deliver viable probiotic cells into gut protectively against harsh environmental conditions of stomach for extended period. Core mucoadhesive microspheres of Bacillus coagulans were prepared using hypromellose, following coacervation and phase separation technique and were then coated with hypromellose phthalate to achieve their site-specific release. Microspheres were evaluated for percent yield, entrapment efficiency, surface morphology, particle size and size distribution, flow property, swelling property, mucoadhesion property by the in vitro wash-off and the ex vivo mucoadhesive strength tests, in vitro release profile and release kinetic, in vivo probiotic activity, and stability. The values for kinetic constant and regression coefficient of model-dependent approaches and the difference factor, the similarity factor, and the Rescigno index of model-independent approaches were determined for accessing and comparing in vitro performance. Microsphere formulation batches have percent yield value between 56.26% and 69.13% and entrapment efficiency value between 66.95% and 77.89%. Microspheres were coarser with spherical shape having mean particle size from 28.03 to 48.31 μm. In vitro B. coagulans release profile follows zero-order kinetics and depends on the grade of hypromellose and the B. coagulans-to-hypromellose ratio. Experimental microspheres rendered adequate stability to B. coagulans at room temperature. Microspheres had delivered B. coagulans in simulated intestinal condition following zero-order kinetics, protectively in simulated gastric condition, exhibiting appreciable mucoadhesion in intestinal condition, which could be useful to achieve site-specific delivery for extended period.
Rotation of an optically trapped vaterite microsphere measured using rotational Doppler effect
NASA Astrophysics Data System (ADS)
Chen, Xinlin; Xiao, Guangzong; Xiong, Wei; Yang, Kaiyong; Luo, Hui; Yao, Baoli
2018-03-01
The angular velocity of a vaterite microsphere spinning in the optical trap is measured using rotational Doppler effect. The perfectly spherical vaterite microspheres are synthesized via coprecipitation in the presence of silk fibroin nanospheres. When trapped by a circularly polarized beam, the vaterite microsphere is uniformly rotated in the trap center. The probe beams containing two Laguerre-Gaussian beams of opposite topological charge l = ± 7, l = ± 8, and l = ± 9 are illuminated on the spinning vaterite. By analyzing the backscattered light, a frequency shift is observed scaling with the rotation rate of the vaterite microsphere. The multiplicative enhancement of the frequency shift proportion to the topological charge has greatly improved the measurement precision. The reliability and practicability of this approach are verified through varying the topological charge of the probe beam and the trapping laser power. In consideration of the excellent measurement precision of the rotation frequency, this technique might be generally applicable in studying the torsional properties of micro-objects.
Using stroboscopic flow imaging to validate large-scale computational fluid dynamics simulations
NASA Astrophysics Data System (ADS)
Laurence, Ted A.; Ly, Sonny; Fong, Erika; Shusteff, Maxim; Randles, Amanda; Gounley, John; Draeger, Erik
2017-02-01
The utility and accuracy of computational modeling often requires direct validation against experimental measurements. The work presented here is motivated by taking a combined experimental and computational approach to determine the ability of large-scale computational fluid dynamics (CFD) simulations to understand and predict the dynamics of circulating tumor cells in clinically relevant environments. We use stroboscopic light sheet fluorescence imaging to track the paths and measure the velocities of fluorescent microspheres throughout a human aorta model. Performed over complex physiologicallyrealistic 3D geometries, large data sets are acquired with microscopic resolution over macroscopic distances.
Ochiuz, Lacramioara; Peris, José-Esteban
2009-03-01
Microparticles of chitosan (CHT) containing alendronate sodium (AL) were prepared in four drug:polymer ratios (1:1, 1:2, 1:4, 1:6) using the spray drying technique. The efficiency of the method was evaluated by determining production yield (about 70 %) and microencapsulation efficiency, which was almost 100 % in the case of all four of the formulations studied. Particles had a mean size of between 3.6 and 4.6 microm, and a near-spherical shape. The formulations with the highest content of AL (drug:polymer ratio 1:1 and 1:2) showed an asymmetrical distribution of particles, which were larger in size, and had a higher proportion of irregular particles than the other formulations. FT-IR analysis revealed an ionic interaction between AL and CHT. Differential scanning calorimetry and thermogravimetric analysis confirmed the microencapsulation of AL and the increased thermal stability of encapsulated AL. The dissolution profiles of AL from CHT microspheres, at pH values of 1.2 and 6.8, showed a delayed release of AL from microspheres, and the dissolution rate was dependent on the pH and the drug:polymer ratio. It can be concluded that spray drying is a suitable technique for preparing AL-loaded CHT microspheres, and that the drug:polymer ratio can be used to control the rate of AL release from microspheres.
Jiang, Tian-Jia; Guo, Zheng; Liu, Jin-Huai; Huang, Xing-Jiu
2015-08-18
An analytical technique based on electroadsorption and transmission X-ray fluorescence (XRF) for the quantitative determination of arsenic in aqueous solution with ppb-level limits of detection (LOD) is proposed. The approach uses electroadsorption to enhance the sensitivity and LOD of the arsenic XRF response. Amine-functionalized carbonaceous microspheres (NH2-CMSs) are found to be the ideal materials for both the quantitative adsorption of arsenic and XRF analysis due to the basic amine sites on the surface and their noninterference in the XRF spectrum. In electroadsorptive X-ray fluorescence (EA-XRF), arsenic is preconcentrated by a conventional three-electrode system with a positive electricity field around the adsorbents. Then, the quantification of arsenic on the adsorbents is achieved using XRF. The electroadsorption preconcentration can realize the fast transfer of arsenic from the solution to the adsorbents and improve the LOD of conventional XRF compared with directly determining arsenic solution by XRF alone. The sensitivity of 0.09 cnt ppb(-1) is obtained without the interferences from coexisted metal ions in the determination of arsenic, and the LOD is found to be 7 ppb, which is lower than the arsenic guideline value of 10 ppb given by the World Health Organization (WHO). These results demonstrated that XRF coupled with electroadsorption was able to determine trace arsenic in real water sample.
USDA-ARS?s Scientific Manuscript database
A multiplex fluorescence microsphere immunoassay (FMIA) was used to detect bovine and ovine IgM and IgG antibodies to several Rift Valley fever virus (RVFV) proteins, including the major surface glycoprotein, Gn; the nonstructural proteins, NSs and NSm; and the nucleoprotein, N. Target antigens were...
Nanomechanics of biocompatible hollow thin-shell polymer microspheres.
Glynos, Emmanouil; Koutsos, Vasileios; McDicken, W Norman; Moran, Carmel M; Pye, Stephen D; Ross, James A; Sboros, Vassilis
2009-07-07
The nanomechanical properties of biocompatible thin-shell hollow polymer microspheres with approximately constant ratio of shell thickness to microsphere diameter were measured by nanocompression tests in aqueous conditions. These microspheres encapsulate an inert gas and are used as ultrasound contrast agents by releasing free microbubbles in the presence of an ultrasound field as a result of free gas leakage from the shell. The tests were performed using an atomic force microscope (AFM) employing the force-distance curve technique. An optical microscope, on which the AFM was mounted, was used to guide the positioning of tipless cantilevers on top of individual microspheres. We performed a systematic study using several cantilevers with spring constants varying from 0.08 to 2.3 N/m on a population of microspheres with diameters from about 2 to 6 microm. The use of several cantilevers with various spring constants allowed a systematic study of the mechanical properties of the microsphere thin shell at different regimes of force and deformation. Using thin-shell mechanics theory for small deformations, the Young's modulus of the thin wall material was estimated and was shown to exhibit a strong size effect: it increased as the shell became thinner. The Young's modulus of thicker microsphere shells converged to the expected value for the macroscopic bulk material. For high applied forces, the force-deformation profiles showed a reversible and/or irreversible nonlinear behavior including "steps" and "jumps" which were attributed to mechanical instabilities such as buckling events.
Prabhu, Sunil; Sullivan, Jennifer L; Betageri, Guru V
2002-01-01
The objective of our study was to compare the in vitro release kinetics of a sustained-release injectable microsphere formulation of the polypeptide drug, calcitonin (CT), to optimize the characteristics of drug release from poly-(lactide-co-glycolide) (PLGA) copolymer biodegradable microspheres. A modified solvent evaporation and double emulsion technique was used to prepare the microspheres. Release kinetic studies were carried out in silanized tubes and dialysis bags, whereby microspheres were suspended and incubated in phosphate buffered saline, sampled at fixed intervals, and analyzed for drug content using a modified Lowry protein assay procedure. An initial burst was observed whereby about 50% of the total dose of the drug was released from the microspheres within 24 hr and 75% within 3 days. This was followed by a period of slow release over a period of 3 weeks in which another 10-15% of drug was released. Drug release from the dialysis bags was more gradual, and 50% CT was released only after 4 days and 75% after 12 days of release. Scanning electron micrographs revealed spherical particles with channel-like structures and a porous surface after being suspended in an aqueous solution for 5 days. Differential scanning calorimetric studies revealed that CT was present as a mix of amorphous and crystalline forms within the microspheres. Overall, these studies demonstrated that sustained release of CT from PLGA microspheres over a 3-week period is feasible and that release of drug from dialysis bags was more predictable than from tubes.
Pagar, Kunal P; Vavia, Pradeep R
2014-06-01
The poly[La-(Glc-Leu)] copolymer was applied in the present investigation as polymeric carrier to fabricate naltrexone (NTX)-loaded poly[La-(Glc-Leu)] microspheres in the single emulsion solvent evaporation technique for the long-term treatment of alcohol dependence. Newly synthesized poly[La-(Glc-Leu)] copolymer exhibited diminished crystallanity, good biocompatibility and favorable biodegradability to be explored for drug delivery application. Scanning Electron Microscopy study revealed smooth and spherical-shaped NTX-loaded polymeric microspheres with a mean size of 10-90 µm. Influence of various decisive formulation variables such as amount of polymer, stabilizer concentration, homogenization speed, homogenization time, drug loading and organic-to-aqueous phase ratio on particle size, and entrapment efficiency was studied. Differential scanning calorimeter and X-ray diffractometry study confirmed the drug entrapment within polymer matrix into the microsphere environment. In vitro drug release showed the sustained drug release of formulation for the period of 28 d giving biphasic release pattern. Histological examination of NTX-loaded poly[La-(Glc-Leu)] microspheres injected intramuscularly into the thigh muscle of Wistar rats showed minimal inflammatory reaction, demonstrating that NTX-loaded microspheres were biocompatible. Insignificant increase in the serum creatine phosphokinase level (p < 0.05) as compared with the normal value revealed good muscle compatibility of the poly[La-(Glc-Leu)] microsphere system. Biocompatible nature and sustained drug-release action of poly[La-(Glc-Leu)] microspheres may have potential application in depot therapy.
Łada, Wiesława; Iller, Edward; Wawszczak, Danuta; Konior, Marcin; Dziel, Tomasz
2016-10-01
A new technology for the production of radiopharmaceutical (90)Y microspheres in the form of spherical yttrium oxide grains obtained by sol-gel method has been described. The authors present and discuss the results of investigations performed in the development of new production technology of yttrium microspheres and determination of their physic-chemical properties. The final product has the structure of spherical yttrium oxide grains with a diameter 25-100μm, is stable and free from contaminants. Irradiation of 20mg samples of grains with diameter of 20-50μm in the thermal neutron flux of 1.7×10(14)cm(-2)s(-1) at the core of MARIA research nuclear reactor allowed to obtain microspheres labelled with the (90)Y isotope on the way of the nuclear reaction (89)Y(n, ɤ)(90)Y. Specific activity of irradiated microspheres has been determined by application of absolute triple to double coincidence ratio method (TDCR) and has been evaluated at 190MBq/mg Y. (90)Y microspheres prepared by the proposed technique can be regarded as a promising medical material for radioembolization of liver malignancies. Copyright © 2016 Elsevier B.V. All rights reserved.
Synthesis of SiCN@TiO2 core-shell ceramic microspheres via PDCs method
NASA Astrophysics Data System (ADS)
Liu, Hongli; Wei, Ning; Li, Jing; Zhang, Haiyuan; Chu, Peng
2018-02-01
A facile and effective polymer-derived ceramics (PDCs) emulsification-crosslinking-pyrolysis method was developed to fabricate SiCN@TiO2 core-shell ceramic microspheres with polyvinylsilazane (PVSZ) and tetrabutyl titanate (TBT) as precursors. The TBT: PVSZ mass ratios, emulsifier concentrations and the pyrolysis temperature were examined as control parameters to tune the size and morphology of microspheres. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the synthesized SiCN@TiO2 microspheres to be comprised of SiCN core coated with TiO2 crystals, with an average size of 0.88 μm when pyrolyzed at 1400 °C. The analysis of Fourier transform infrared spectroscopy (FT-IR), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) ensured that SiCN@TiO2 core-shell ceramic microspheres composed of rutile TiO2, β-SiC and Si3N4 crystalline phases, The thermal properties were characterized by thermogravimetric analysis (TGA). The obtained SiCN@TiO2 core-shell ceramic microspheres were the promising candidate of the infrared opacifier in silica aerogels and this technique can be extended to other preceramic polymers.
Cho, Yi Je; Lee, Wookjin; Park, Yong Ho
2017-01-01
The elastoplastic deformation behaviors of hollow glass microspheres/iron syntactic foam under tension were modeled using a representative volume element (RVE) approach. The three-dimensional microstructures of the iron syntactic foam with 5 wt % glass microspheres were reconstructed using the random sequential adsorption algorithm. The constitutive behavior of the elastoplasticity in the iron matrix and the elastic-brittle failure for the glass microsphere were simulated in the models. An appropriate RVE size was statistically determined by evaluating elastic modulus, Poisson’s ratio, and yield strength in terms of model sizes and boundary conditions. The model was validated by the agreement with experimental findings. The tensile deformation mechanism of the syntactic foam considering the fracture of the microspheres was then investigated. In addition, the feasibility of introducing the interfacial deboning behavior to the proposed model was briefly investigated to improve the accuracy in depicting fracture behaviors of the syntactic foam. It is thought that the modeling techniques and the model itself have major potential for applications not only in the study of hollow glass microspheres/iron syntactic foams, but also for the design of composites with a high modulus matrix and high strength reinforcement. PMID:29048346
A sensitive pressure sensor for diamond anvil cell experiments up to 2 GPa: FluoSpheres[reg
DOE Office of Scientific and Technical Information (OSTI.GOV)
Picard, Aude; Oger, Phil M.; Daniel, Isabelle
2006-08-01
We present an optical pressure sensor suitable for experiments in diamond anvil cell in the 0.1 MPa-2 GPa pressure range, for temperatures between ambient and 323 K. It is based on the pressure-dependent fluorescence spectrum of FluoSpheres[reg], which are commercially available fluorescent microspheres commonly used to measure blood flow in experimental biology. The fluorescence of microspheres is excited by the 514.5 nm line of an Ar{sup +} laser, and the resulting spectrum displays three very intense broad bands at 534, 558, and 598 nm, respectively. The reference wavelength and pressure gauge is that of the first inflection point of themore » spectrum, located at 525.6{+-}0.2 nm at ambient pressure. It is characterized by an instantaneous and large linear pressure shift of 9.93({+-}0.08) nm/GPa. The fluorescence of the FluoSpheres[reg] has been investigated as a function of pressure (0.1-4 GPa), temperature (295-343 K), pH (3-12), salinity, and pressure transmitting medium. These measurements show that, for pressures comprised between 0.1 MPa and 2 GPa, at temperatures not exceeding 323 K, at any pH, in aqueous pressure transmitting media, pressure can be calculated from the wavelength shift of two to three beads, according to the relation P=0.100 ({+-}0.001) {delta}{lambda}{sub i}(P) with {delta}{lambda}{sub i}(P)={lambda}{sub i}(P)-{lambda}{sub i}(0) and {lambda}{sub i}(P) as the wavelength of the first inflection point of the spectrum at the pressure P. This pressure sensor is approximately thirty times more sensitive than the ruby scale and responds instantaneously to pressure variations.« less
Wygladacz, Katarzyna; Radu, Aleksandar; Xu, Chao; Qin, Yu; Bakker, Eric
2005-08-01
An optical microsensor array is described for the rapid analysis of silver ions at low parts per trillion levels. Because the ionophore o-xylylenebis(N,N-diisobutyldithiocarbamate) (Cu-I) was reevaluated and shown to exhibit excellent selectivity for silver ions, ion-selective electrode (ISE) membranes were optimized and found to exhibit the lowest reported detection limit so far (3 x 10(-10) M). A corresponding Ag+-selective fluorescent optical microsensor array for the rapid sensing of trace level Ag+ was then developed. It was fabricated using plasticized PVC-based micrometer-scale fluorescent microspheres that were produced via a sonic particle casting device. They contained 156 mmol/kg Cu-I, 10 mmol/kg 9-(diethylamino)-5-[4-(15-butyl-1,13-dioxo-2,14-dioxanodecyl) phenylimino]benzo[a]phenoxazine (chromoionophore VII, ETH 5418), 2.3 mmol/kg 1,1' '-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (internal reference dye), and 14 mmol/kg sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate and were deposited onto the etched distal end of a 3200-microm-diameter optical fiber bundle. The microarray was characterized by fluorescence spectroscopy in samples containing 10(-12)-10(-8) M AgNO3 at pH 7.4, with selectivity characteristics comparable to the corresponding ISEs. The response time of the microsensor array was found to be less than 15 min for 10(-9) M AgNO3, which is drastically shorter than earlier data on optode films (8 h) and corresponding ISEs (30 min). A detection limit of 4 x 10(-11) M for Ag+ was observed, lower than any previously reported optode or silver-selective ISE. The microsensor array was applied for measurement of free silver levels in buffered pond water samples.
Nicholas, Matthew P.; Rao, Lu; Gennerich, Arne
2014-01-01
Rigid attachment of microtubules (MTs) to glass cover slip surfaces is a prerequisite for a variety of microscopy experiments in which MTs are used as substrates for MT-associated proteins, such as the molecular motors kinesin and cytoplasmic dynein. We present an MT-surface coupling protocol in which aminosilanized glass is formylated using the cross-linker glutaraldehyde, fluorescence-labeled MTs are covalently attached, and the surface is passivated with highly pure beta-casein. The technique presented here yields rigid MT immobilization while simultaneously blocking the remaining glass surface against nonspecific binding by polystyrene optical trapping microspheres. This surface chemistry is straightforward and relatively cheap and uses a minimum of specialized equipment or hazardous reagents. These methods provide a foundation for a variety of optical tweezers experiments with MT-associated molecular motors and may also be useful in other assays requiring surface-immobilized proteins. PMID:24633798
Xiong, Long; Zeng, Jianhua; Yao, Aihua; Tu, Qiquan; Li, Jingtang; Yan, Liang; Tang, Zhiming
2015-01-01
The regeneration of large bone defects is an osteoinductive, osteoconductive, and osteogenic process that often requires a bone graft for support. Limitations associated with naturally autogenic or allogenic bone grafts have demonstrated the need for synthetic substitutes. The present study investigates the feasibility of using novel hollow hydroxyapatite microspheres as an osteoconductive matrix and a carrier for controlled local delivery of bone morphogenetic protein 2 (BMP2), a potent osteogenic inducer of bone regeneration. Hollow hydroxyapatite microspheres (100±25 μm) with a core (60±18 μm) and a mesoporous shell (180±42 m(2)/g surface area) were prepared by a glass conversion technique and loaded with recombinant human BMP2 (1 μg/mg). There was a gentle burst release of BMP2 from microspheres into the surrounding phosphate-buffered saline in vitro within the initial 48 hours, and continued at a low rate for over 40 days. In comparison with hollow hydroxyapatite microspheres without BMP2 or soluble BMP2 without a carrier, BMP2-loaded hollow hydroxyapatite microspheres had a significantly enhanced capacity to reconstitute radial bone defects in rabbit, as shown by increased serum alkaline phosphatase; quick and complete new bone formation within 12 weeks; and great biomechanical flexural strength. These results indicate that BMP2-loaded hollow hydroxyapatite microspheres could be a potential new option for bone graft substitutes in bone regeneration.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Lau, Wan-Yee, E-mail: josephlau@surgery.cuhk.edu.hk; Kennedy, Andrew S.; Department of Biomedical Engineering, North Carolina State University, Raleigh, NC
Purpose: Selective internal radiotherapy (SIRT) with yttrium-90 ({sup 90}Y) resin microspheres can improve the clinical outcomes for selected patients with inoperable liver cancer. This technique involves intra-arterial delivery of {beta}-emitting microspheres into hepatocellular carcinomas or liver metastases while sparing uninvolved structures. Its unique mode of action, including both {sup 90}Y brachytherapy and embolization of neoplastic microvasculature, necessitates activity planning methods specific to SIRT. Methods and Materials: A panel of clinicians experienced in {sup 90}Y resin microsphere SIRT was convened to integrate clinical experience with the published data to propose an activity planning pathway for radioembolization. Results: Accurate planning is essentialmore » to minimize potentially fatal sequelae such as radiation-induced liver disease while delivering tumoricidal {sup 90}Y activity. Planning methods have included empiric dosing according to degree of tumor involvement, empiric dosing adjusted for the body surface area, and partition model calculations using Medical Internal Radiation Dose principles. It has been recommended that at least two of these methods be compared when calculating the microsphere activity for each patient. Conclusions: Many factors inform {sup 90}Y resin microsphere SIRT activity planning, including the therapeutic intent, tissue and vasculature imaging, tumor and uninvolved liver characteristics, previous therapies, and localization of the microsphere infusion. The influence of each of these factors has been discussed.« less
Single-analyte to multianalyte fluorescence sensors
NASA Astrophysics Data System (ADS)
Lavigne, John J.; Metzger, Axel; Niikura, Kenichi; Cabell, Larry A.; Savoy, Steven M.; Yoo, J. S.; McDevitt, John T.; Neikirk, Dean P.; Shear, Jason B.; Anslyn, Eric V.
1999-05-01
The rational design of small molecules for the selective complexation of analytes has reached a level of sophistication such that there exists a high degree of prediction. An effective strategy for transforming these hosts into sensors involves covalently attaching a fluorophore to the receptor which displays some fluorescence modulation when analyte is bound. Competition methods, such as those used with antibodies, are also amenable to these synthetic receptors, yet there are few examples. In our laboratories, the use of common dyes in competition assays with small molecules has proven very effective. For example, an assay for citrate in beverages and an assay for the secondary messenger IP3 in cells have been developed. Another approach we have explored focuses on multi-analyte sensor arrays with attempt to mimic the mammalian sense of taste. Our system utilizes polymer resin beads with the desired sensors covalently attached. These functionalized microspheres are then immobilized into micromachined wells on a silicon chip thereby creating our taste buds. Exposure of the resin to analyte causes a change in the transmittance of the bead. This change can be fluorescent or colorimetric. Optical interrogation of the microspheres, by illuminating from one side of the wafer and collecting the signal on the other, results in an image. These data streams are collected using a CCD camera which creates red, green and blue (RGB) patterns that are distinct and reproducible for their environments. Analysis of this data can identify and quantify the analytes present.
Långmark, Jonas; Storey, Michael V.; Ashbolt, Nicholas J.; Stenström, Thor-Axel
2005-01-01
The accumulation and fate of model microbial “pathogens” within a drinking-water distribution system was investigated in naturally grown biofilms formed in a novel pilot-scale water distribution system provided with chlorinated and UV-treated water. Biofilms were exposed to 1-μm hydrophilic and hydrophobic microspheres, Salmonella bacteriophages 28B, and Legionella pneumophila bacteria, and their fate was monitored over a 38-day period. The accumulation of model pathogens was generally independent of the biofilm cell density and was shown to be dependent on particle surface properties, where hydrophilic spheres accumulated to a larger extent than hydrophobic ones. A higher accumulation of culturable legionellae was measured in the chlorinated system compared to the UV-treated system with increasing residence time. The fate of spheres and fluorescence in situ hybridization-positive legionellae was similar and independent of the primary disinfectant applied and water residence time. The more rapid loss of culturable legionellae compared to the fluorescence in situ hybridization-positive legionellae was attributed to a loss in culturability rather than physical desorption. Loss of bacteriophage 28B plaque-forming ability together with erosion may have affected their fate within biofilms in the pilot-scale distribution system. The current study has demonstrated that desorption was one of the primary mechanisms affecting the loss of microspheres, legionellae, and bacteriophage from biofilms within a pilot-scale distribution system as well as disinfection and biological grazing. In general, two primary disinfection regimens (chlorination and UV treatment) were not shown to have a measurable impact on the accumulation and fate of model microbial pathogens within a water distribution system. PMID:15691920
Långmark, Jonas; Storey, Michael V; Ashbolt, Nicholas J; Stenström, Thor-Axel
2005-02-01
The accumulation and fate of model microbial "pathogens" within a drinking-water distribution system was investigated in naturally grown biofilms formed in a novel pilot-scale water distribution system provided with chlorinated and UV-treated water. Biofilms were exposed to 1-mum hydrophilic and hydrophobic microspheres, Salmonella bacteriophages 28B, and Legionella pneumophila bacteria, and their fate was monitored over a 38-day period. The accumulation of model pathogens was generally independent of the biofilm cell density and was shown to be dependent on particle surface properties, where hydrophilic spheres accumulated to a larger extent than hydrophobic ones. A higher accumulation of culturable legionellae was measured in the chlorinated system compared to the UV-treated system with increasing residence time. The fate of spheres and fluorescence in situ hybridization-positive legionellae was similar and independent of the primary disinfectant applied and water residence time. The more rapid loss of culturable legionellae compared to the fluorescence in situ hybridization-positive legionellae was attributed to a loss in culturability rather than physical desorption. Loss of bacteriophage 28B plaque-forming ability together with erosion may have affected their fate within biofilms in the pilot-scale distribution system. The current study has demonstrated that desorption was one of the primary mechanisms affecting the loss of microspheres, legionellae, and bacteriophage from biofilms within a pilot-scale distribution system as well as disinfection and biological grazing. In general, two primary disinfection regimens (chlorination and UV treatment) were not shown to have a measurable impact on the accumulation and fate of model microbial pathogens within a water distribution system.
High-density fiber optic biosensor arrays
NASA Astrophysics Data System (ADS)
Epstein, Jason R.; Walt, David R.
2002-02-01
Novel approaches are required to coordinate the immense amounts of information derived from diverse genomes. This concept has influenced the expanded role of high-throughput DNA detection and analysis in the biological sciences. A high-density fiber optic DNA biosensor was developed consisting of oligonucleotide-functionalized, 3.1 mm diameter microspheres deposited into the etched wells on the distal face of a 500 micrometers imaging fiber bundle. Imaging fiber bundles containing thousands of optical fibers, each associated with a unique oligonucleotide probe sequence, were the foundation for an optically connected, individually addressable DNA detection platform. Different oligonucleotide-functionalized microspheres were combined in a stock solution, and randomly dispersed into the etched wells. Microsphere positions were registered from optical dyes incorporated onto the microspheres. The distribution process provided an inherent redundancy that increases the signal-to-noise ratio as the square root of the number of sensors examined. The representative amount of each probe-type in the array was dependent on their initial stock solution concentration, and as other sequences of interest arise, new microsphere elements can be added to arrays without altering the existing detection capabilities. The oligonucleotide probe sequences hybridize to fluorescently-labeled, complementary DNA target solutions. Fiber optic DNA microarray research has included DNA-protein interaction profiles, microbial strain differentiation, non-labeled target interrogation with molecular beacons, and single cell-based assays. This biosensor array is proficient in DNA detection linked to specific disease states, single nucleotide polymorphism (SNP's) discrimination, and gene expression analysis. This array platform permits multiple detection formats, provides smaller feature sizes, and enables sensor design flexibility. High-density fiber optic microarray biosensors provide a fast, reversible format with the detection limit of a few hundred molecules.
Plasmonic improvement of microcavity biomedical sensor spectroscopic characteristics
NASA Astrophysics Data System (ADS)
Saetchnikov, Vladimir A.; Tcherniavskaia, Elina A.; Saetchnikov, Anton V.; Schweiger, Gustav; Ostendorf, Andreas; Ghadiri, Reza
2014-03-01
New opportunity to improve a sensetivity of a label-free biomolecule detection in sensing systems based on microcavity evanescent wave optical sensors has been recently found and is being under intensive development. Novel technique based on combination of optical resonance on microring structures with plasmon resonance. Recently developed tools based on neural network data processing can realize real-time identification of biological agents. So combining advantages of plasmon enhancing optical microcavity resonance with identification tools can give a new platform for ulta sensitive label-free biomedical sensor. Our developed technique used standard glass and polymer microspheres as sensetive elements. They are fixed in the solution flow by adhesive layer on the surface being in the field of evanescence wave. Sensitive layer have been treated by gold nanoparticel (GN) solution. Another technique used thin film gold layers deposited on the substrate below adhesive. The light from a tuneable diode laser is coupled into the microsphere through a prism and was sharply focussed on the single microsphere. Images were recorded by CMOS camera. Normalized by free spectral range resonance shift of whispering gallery mode (WGM) and a relative efficiency of their excitation were used as input data for biomolecule classification. Both biomolecules and NP injection was obtained caused WGM spectra modification. But after NP treatment spectral shift and intensity of WGM resonances in biomolecule solutions increased. WGM resonances in microspheres fixed on substrate with gold layer with optimized layer thickness in biomolecule solutions also had higher intensity and spectra modification then without gold layer.
Oz, Umut Can; Devrim, Burcu; Bozkır, Asuman; Canefe, Kandemir
2015-01-01
Effective clinical utilisation of non-steroidal anti-inflammatory drugs, such as diclofenac sodium (DS) is significantly limited by their ulcerogenic potential and poor bioavailability after oral administration. The objective of this work was to develop reconstitutable pediatric suspensions of DS-loaded microspheres prepared with an acrylic polymer (Eudragit RS) for improved pediatric delivery of DS. The microspheres were prepared by the water-in-oil-in-water or solid-in-oil-in-water emulsion techniques. Enviromental scanning electron microscopy observations clearly showed that microspheres have spherical shape. The drug entrapment efficiency of these microspheres was found 47.96 ± 0.79% to 88.57 ± 0.59% and their average particle sizes were 23.94-60.78 µm, which are within the desired range for the development of suspension formulation. The in vitro dissolution indicated prolonged sustained release of DS over 8 h. The results of preliminary characterisation studies of suspensions show that a liquid pharmaceutical preparation for oral administration capable of providing a sustained release of DS was successfully obtained.
Soft tissue augmentation with artecoll: 10-year history, indications, techniques, and complications.
Lemperle, Gottfried; Romano, James J; Busso, Mariano
2003-06-01
Most of the biologic filler materials that increase the thickness of the corium in a wrinkle line are phagocytosed within a certain time. Therefore, a lasting effect can only be achieved with nonresorbable synthetic substances. Artefill consists of 20 volume percent microspheres of polymethyl-methacrylate and 80 volume percent of bovine collagen. Beneath the crease, the microspheres with their exceptional surface smoothness stimulate fibroblasts to encapsulate each individual one of the 6-million microspheres contained in 1 mL of Artefill. Collagen is merely a carrier substance that prevents the microspheres from agglomerating during tissue ingrowth. The 20 volume percent of microspheres in Artefill provides the scaffold for the 80% volume of connective tissue deposition, a complete replacement of the injected collagen. The filler material beneath a crease acts like a splint and prevents the possibility of its further folding, thereby allowing the diminished thickness of the corium in a crease to recover. This recovery process is well known even in older patients with facial paralysis or after a stroke, whose facial wrinkles and furrows on the paralyzed side disappear over time.
Removal of radioactive contaminants by polymeric microspheres.
Osmanlioglu, Ahmet Erdal
2016-11-01
Radionuclide removal from radioactive liquid waste by adsorption on polymeric microspheres is the latest application of polymers in waste management. Polymeric microspheres have significant immobilization capacity for ionic substances. A laboratory study was carried out by using poly(N-isopropylacrylamide) for encapsulation of radionuclide in the liquid radioactive waste. There are numbers of advantages to use an encapsulation technology in radioactive waste management. Results show that polymerization step of radionuclide increases integrity of solidified waste form. Test results showed that adding the appropriate polymer into the liquid waste at an appropriate pH and temperature level, radionuclide was encapsulated into polymer. This technology may provide barriers between hazardous radioactive ions and the environment. By this method, solidification techniques became easier and safer in nuclear waste management. By using polymer microspheres as dust form, contamination risks were decreased in the nuclear industry and radioactive waste operations.
Sóñora, Cecilia; Arbildi, Paula; Miraballes-Martínez, Iris; Hernández, Ana
2018-01-01
Phagocytosis is a fundamental process for removal of pathogens and for clearance of apoptotic cells. The objective of this work was the preparation of fluorescent microspheres by a simple method and the evaluation of its applicability in phagocytosis assays by using different human derived cells, differentiated THP-1 cell line and blood monocytes, with flow cytometry measurements for functionality assays. Our results show that microparticles are efficiently internalised in a non-opsonised form and in dose-dependent manner by both cellular types. Concerning mechanism we determined that tTG-β3 integrin signaling could be involved in the uptake of these particles.
Applications of optical manipulation in plant biology
NASA Astrophysics Data System (ADS)
Buer, Charles S.
Measuring small forces in biology is important for determining basic physiological parameters of a cell. The plant cell wall provides a primary defense and presents a barrier to research. Magnitudes of small forces are impossible to measure with mechanical transducers, glass needles, atomic force microscopy, or micropipet-based force transduction due to the cell wall. Therefore, a noninvasive method of breaching the plant cell wall to access the symplastic region of the cell is required. Laser light provides sub-micrometer positioning, particle manipulation without mechanical contact, and piconewton force determination. Consequently, the extension of laser microsurgery to expand an experimental tool for plant biology encompassed the overall objective. A protocol was developed for precisely inserting microscopic objects into the periplasmic region of plant callus cells using laser microsurgery. Ginkgo biloba and Agrobacterium rhizogenes were used as the model system for developing the optical tweezers and scalpel techniques. Better than 95% survival was achieved after plasmolyzing G. biloba cells, ablating a 2-4 μm hole through the cell wall using a pulsed UV laser beam, trapping and manipulating bacteria into the periplasmic region, and deplasmolyzing the cells. Optical trapping experiments implied a difference existed between the bacteria models. Determining the optical trapping efficiency of Agrobacterium rhizogenes and A. tumefaciens strains indicated the A. rhizogenes strain, ATCC 11325, was significantly less efficiently trapped than strains A4 and ATCC 15834 and the A. tumefaciens strain LBA4404. Differences were also found in capsule generation, growth media viscosity, and transmission electron microscopy negative staining implying that a difference in surface structure exists. Calcofluor fluorescence suggests the difference involves an exopolysaccharide. Callus cell plasmolysis revealed Hechtian strands interconnecting the plasma membrane and the cell wall. The spring tension of these strands was measured in normal and cold-hardened G. biloba and N. tabacum callus cells. There was little change in flexibility between the groups of cultured cells in either species studied. Microspheres were attached to Hechtian strands in normal cultured Nicotiana tabacum and the cells were deplasmolyzed and replasmolyzed to determine the fate of Hechtian strands. The microspheres either moved to the plasma membrane and adhered or moved to the cell wall and adhered. The attached microspheres occasionally moved independently on the same strand. Inserted microspheres provided a visual probe to follow physiological events within a plant cell.
Differential white cell count by centrifugal microfluidics.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sommer, Gregory Jon; Tentori, Augusto M.; Schaff, Ulrich Y.
We present a method for counting white blood cells that is uniquely compatible with centrifugation based microfluidics. Blood is deposited on top of one or more layers of density media within a microfluidic disk. Spinning the disk causes the cell populations within whole blood to settle through the media, reaching an equilibrium based on the density of each cell type. Separation and fluorescence measurement of cell types stained with a DNA dye is demonstrated using this technique. The integrated signal from bands of fluorescent microspheres is shown to be proportional to their initial concentration in suspension. Among the current generationmore » of medical diagnostics are devices based on the principle of centrifuging a CD sized disk functionalized with microfluidics. These portable 'lab on a disk' devices are capable of conducting multiple assays directly from a blood sample, embodied by platforms developed by Gyros, Samsung, and Abaxis. [1,2] However, no centrifugal platform to date includes a differential white blood cell count, which is an important metric complimentary to diagnostic assays. Measuring the differential white blood cell count (the relative fraction of granulocytes, lymphocytes, and monocytes) is a standard medical diagnostic technique useful for identifying sepsis, leukemia, AIDS, radiation exposure, and a host of other conditions that affect the immune system. Several methods exist for measuring the relative white blood cell count including flow cytometry, electrical impedance, and visual identification from a stained drop of blood under a microscope. However, none of these methods is easily incorporated into a centrifugal microfluidic diagnostic platform.« less
Effective cell trapping using PDMS microspheres in an acoustofluidic chip.
Yin, Di; Xu, Gangwei; Wang, Mengyuan; Shen, Mingwu; Xu, Tiegang; Zhu, Xiaoyue; Shi, Xiangyang
2017-09-01
We present a facile particle-based cell manipulation method using acoustic radiation forces. In this work, we selected several representative particles including poly(lactic-co-glycolic acid) (PLGA) microspheres, silica-coated magnetic microbeads, polydimethylsiloxane (PDMS) microspheres and investigated the responses of these particle systems to ultrasonic standing waves (USWs) in a microfluidic chip. We show that depending on the nature (positive or negative acoustic contrast factors) of the particles, these particle systems display different alignment behaviors along the microfluidic channel under USWs. Specifically, PLGA microspheres and silica-coated magnetic microbeads are able to be aligned in the middle of the microfluidic channel, while PDMS microspheres are translocated to the side walls of the channel, which is beneficial for cell trapping and manipulation. Further results demonstrate that the functional PDMS microspheres with a negative acoustic contrast factor can be used to trap cells to the pressure antinodes in the acoustofluidic chip. Cell viability tests reveal that the ultrasonic manipulation does not exert any harmful effect to the cells. This acoustic-based particle and cell manipulation technique may hold a great promise for the development of rapid, noninvasive, continuous assays for detecting of cells and separation of biological samples. Copyright © 2017 Elsevier B.V. All rights reserved.
Hosseini, Samira; Aeinehvand, Mohammad M.; Uddin, Shah M.; Benzina, Abderazak; Rothan, Hussin A.; Yusof, Rohana; Koole, Leo H.; Madou, Marc J.; Djordjevic, Ivan; Ibrahim, Fatimah
2015-01-01
The application of microfluidic devices in diagnostic systems is well-established in contemporary research. Large specific surface area of microspheres, on the other hand, has secured an important position for their use in bioanalytical assays. Herein, we report a combination of microspheres and microfluidic disk in a unique hybrid platform for highly sensitive and selective detection of dengue virus. Surface engineered polymethacrylate microspheres with carefully designed functional groups facilitate biorecognition in a multitude manner. In order to maximize the utility of the microspheres’ specific surface area in biomolecular interaction, the microfluidic disk was equipped with a micromixing system. The mixing mechanism (microballoon mixing) enhances the number of molecular encounters between spheres and target analyte by accessing the entire sample volume more effectively, which subsequently results in signal amplification. Significant reduction of incubation time along with considerable lower detection limits were the prime motivations for the integration of microspheres inside the microfluidic disk. Lengthy incubations of routine analytical assays were reduced from 2 hours to 5 minutes while developed system successfully detected a few units of dengue virus. Obtained results make this hybrid microsphere-microfluidic approach to dengue detection a promising avenue for early detection of this fatal illness. PMID:26548806
Wei, Qiang; Wei, Wei; Tian, Rui; Wang, Lian-Yan; Su, Zhi-Guo; Ma, Guang-Hui
2008-07-15
Relatively uniform-sized poly(lactide-co-ethylene glycol) (PELA) microspheres with high encapsulation efficiency were prepared rapidly by a novel method combining emulsion-solvent extraction and premix membrane emulsification. Briefly, preparation of coarse double emulsions was followed by additional premix membrane emulsification, and antigen-loaded microspheres were obtained by further solidification. Under the optimum condition, the particle size was about 1 mum and the coefficient of variation (CV) value was 18.9%. Confocal laser scanning microscope and flow cytometer analysis showed that the inner droplets were small and evenly dispersed and the antigen was loaded uniformly in each microsphere when sonication technique was occupied to prepare primary emulsion. Distribution pattern of PEG segment played important role on the properties of microspheres. Compared with triblock copolymer PLA-PEG-PLA, the diblock copolymer PLA-mPEG yielded a more stable interfacial layer at the interface of oil and water phase, and thus was more suitable to stabilize primary emulsion and protect coalescence of inner droplets and external water phase, resulting in high encapsulation efficiency (90.4%). On the other hand, solidification rate determined the time for coalescence during microspheres fabrication, and thus affected encapsulation efficiency. Taken together, improving the polymer properties and solidification rate are considered as two effective strategies to yield high encapsulation.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Hubbard, L; Ziemer, B; Sadeghi, B
Purpose: To evaluate the accuracy of dynamic CT myocardial perfusion measurement using first pass analysis (FPA) and maximum slope models. Methods: A swine animal model was prepared by percutaneous advancement of an angioplasty balloon into the proximal left anterior descending (LAD) coronary artery to induce varying degrees of stenosis. Maximal hyperaemia was achieved in the LAD with an intracoronary adenosine drip (240 µg/min). Serial microsphere and contrast (370 mg/mL iodine, 30 mL, 5mL/s) injections were made over a range of induced stenoses, and dynamic imaging was performed using a 320-row CT scanner at 100 kVp and 200 mA. The FPAmore » CT perfusion technique was used to make vessel-specific myocardial perfusion measurements. CT perfusion measurements using the FPA and maximum slope models were validated using colored microspheres as the reference gold standard. Results: Perfusion measurements using the FPA technique (P-FPA) showed good correlation with minimal offset when compared to perfusion measurements using microspheres (P- Micro) as the reference standard (P -FPA = 0.96 P-Micro + 0.05, R{sup 2} = 0.97, RMSE = 0.19 mL/min/g). In contrast, the maximum slope model technique (P-MS) was shown to underestimate perfusion when compared to microsphere perfusion measurements (P-MS = 0.42 P -Micro −0.48, R{sup 2} = 0.94, RMSE = 3.3 mL/min/g). Conclusion: The results indicate the potential for significant improvements in accuracy of dynamic CT myocardial perfusion measurement using the first pass analysis technique as compared with the standard maximum slope model.« less
NASA Astrophysics Data System (ADS)
Li, Joanne; Dobrucki, Lawrence W.; Marjanovic, Marina; Chaney, Eric J.; Suslick, Kenneth S.; Boppart, Stephen A.
2015-01-01
Cerenkov luminescence (CL) imaging is a new molecular imaging modality that utilizes the photons emitted during radioactive decay when charged particles travel faster than the phase velocity of light in a dielectric medium. Here we present a novel agent to convert and increase CL emission at longer wavelengths using multimodal protein microspheres (MSs). The 64Cu-labeled protein microspheres contain quantum dots (QDs) encapsulated within a high-refractive-index-oil core. Dark box imaging of the MSs was conducted to demonstrate the improvement in CL emission at longer wavelengths. To illustrate the versatile design of these MSs and the potential of CL in disease diagnosis, these MSs were utilized for in vitro cell targeting and ex vivo CL-excited QD fluorescence (CL-FL) imaging of atherosclerotic plaques in rats. It was shown that by utilizing both QDs and MSs with a high-refractive-index-oil core, the CL emission increases by four-fold at longer wavelengths. Furthermore, we demonstrate that these MSs generate both an in vivo and ex vivo contrast signal. The design concept of utilizing QDs and high-index core MSs may contribute to future developments of in vivo CL imaging.
[Building immune microsphere against tumor necrosis factor-alpha (TNF-alpha)].
Wang, Qin; Wu, Xiongfei; Wang, Junxia; Liu, Hong; Li, Lian; Jin, Xiyu
2005-12-01
We have constructed the immune microsphere against tumor necrosis factor-alpha (TNF-alpha) prospectively, hoping to establish the experiment groundwork in more researches which could be used in specific elimination of the TNF-alpha by blood purification method for the future. The recombinant human tumor necrosis factor-alpha monoclonal antibody (rHTNF-alpha McAb) was wrapped on the polystyrene microsphere (PSM) carrier connecting poly-L-lysine (PLL) beforehand. They were earmarked by the fluorescein isothiocyanate (FITC) respectively. The packing conditions were examined using the inversted and fluorescence microscopes and the spectrophotometer. The results showed that the best conditions for wrapping were 20 degrees C, pH9.5 and 60 minutes. The PLL content was not changed in the washing fluid after coating, which indicated the wrapping was quite firm. At the same temperature and same coating time, the rHTNF-alpha McAb coated on the PLL was obviously substantial when the concentration of glutaraldehyde solution was 0.2%. The findings demonstrated that the built immune microsphers can be used as a novel adsorption material. This method is simple and economic, and it offers a new approach to the related studies.
NASA Astrophysics Data System (ADS)
Savoy, Steven M.; Lavigne, John J.; Yoo, J. S.; Wright, John; Rodriguez, Marc; Goodey, Adrian; McDoniel, Bridget; McDevitt, John T.; Anslyn, Eric V.; Shear, Jason B.; Ellington, Andrew D.; Neikirk, Dean P.
1998-12-01
A micromachined sensor array has been developed for the rapid characterization of multi-component mixtures in aqueous media. The sensor functions in a manner analogous to that of the mammalian tongue, using an array composed of individually immobilized polystyrene-polyethylene glycol composite microspheres selectively arranged in micromachined etch cavities localized o n silicon wafers. Sensing occurs via colorimetric or fluorometric changes to indicator molecules that are covalently bound to amine termination sites on the polymeric microspheres. The hybrid micromachined structure has been interfaced directly to a charged-coupled-device that is used for the simultaneous acquisition of the optical data from the individually addressable `taste bud' elements. With the miniature sensor array, acquisition of data streams composed of red, green, and blue color patterns distinctive for the analytes in the solution are rapidly acquired. The unique combination of carefully chosen reporter molecules with water permeable microspheres allows for the simultaneous detection and quantification of a variety of analytes. The fabrication of the sensor structures and the initial colorimetric and fluorescent responses for pH, Ca+2, Ce+3, and sugar are reported. Interface to microfluidic components should also be possible, producing a complete sampling/sensing system.
[Preparation of scopolamine hydrobromide nanoparticles-in-microsphere system].
Lü, Wei-ling; Hu, Jin-hong; Zhu, Quan-gang; Li, Feng-qian
2010-07-01
This study is to prepare scopolamine hydrobromide nanoparticles-in-microsphere system (SH-NiMS) and evaluate its drug release characteristics in vitro. SH nanoparticles were prepared by ionic crosslinking method with tripolyphosphate (TPP) as crosslinker and chitosan as carrier. Orthogonal design was used to optimize the formulation of SH nanoparticles, which took the property of encapsulation efficiency and drug loading as evaluation parameters. With HPMC as carrier, adjusted the parameters of spray drying technique and sprayed the SH nanoparticles in microspheres encaposulated by HPMC was formed and which is called nanoparticles-in-microsphere system (NiMS). SH-NiMS appearances were observed by SEM, structure was obsearved by FT-IR and the release characteristics in vitro were evaluated. The optimized formulation of SH nanoparticles was TPP/CS 1:3 (w/w), HPMC 0.3%, SH 0.2%. The solution peristaltic speed of the spray drying technique was adjusted to 15%, and the temperature of inlet was 110 degrees C. The encapsulation product yeild, drug loading and particle sizes of SH-NiMS were 94.2%, 20.4%, and 1256.5 nm, respectively. The appearances and the structure of SH-NiMS were good. The preparation method of SH-NiMS is stable and reliable to use, which provide a new way to develop new dosage form.
Yang, Fan; Chen, De; Guo, Zhe-Fei; Zhang, Yong-Ming; Liu, Yi; Askin, Sean; Craig, Duncan Q M; Zhao, Min
2017-04-30
Poly (d,l-lactic-co-glycolic) acid (PLGA) based microspheres have been extensively used as controlled drug release systems. However, the burst effect has been a persistent issue associated with such systems, especially for those prepared by the double emulsion technique. An effective approach to preventing the burst effect and achieving a more ideal drug release profile is to improve the drug distribution within the polymeric matrix. Therefore, it is of great importance to establish a rapid and robust tool for screening and optimizing the drug distribution during pre-formulation. Transition Temperature Microscopy (TTM), a novel nano-thermal and imaging technique, is an extension of nano-thermal analysis (nano-TA) whereby a transition temperature is detected at a localized region of a sample and then designated a color based on a particular temperature/color palette, finally resulting in a coded map based on transition temperatures detected by carrying out a series of nanoTA measurements across the surface of the sample. In this study, we investigate the feasibility of applying the aforementioned technique combined with other thermal, imaging and structural techniques for monitoring the drug microstructure and spatial distribution within bovine serum albumin (BSA) loaded and nimodipine loaded PLGA microspheres, with a view to better predicting the in vitro drug release performance. Copyright © 2017 Elsevier B.V. All rights reserved.
Wei, Yi; Wang, Yuxia; Kang, Aijun; Wang, Wei; Ho, Sa V; Gao, Junfeng; Ma, Guanghui; Su, Zhiguo
2012-07-02
An effective and safe formulation of sustained-release rhGH for two months using poly(monomethoxypolyethylene glycol-co-D,L-lactide) (mPEG-PLA, PELA) microspheres was developed to reduce the frequency of medication. The rhGH-loaded PELA microspheres with a narrow size distribution were successfully prepared by a double emulsion method combined with a premix membrane emulsification technique without any exogenous stabilizing excipients. The narrow size distribution of the microspheres would guarantee repeatable productivity and release behavior. Moreover, the amphiphilic PELA improved the bioactivity retention of protein drugs since it prevented protein contact with the oil/water interface and the hydrophobic network, and modulated diffusion of acidic degradation products from the carrier system. These PELA microspheres were compared in vivo with commercial rhGH solution, conventional poly(D,L-lactic acid) (PLA) and poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres. Administration of rhGH-PELA could extend the duration of rhGH release (for up to 56 days) and increase area under the curve (AUC) compared to rhGH solution, PLA or PLGA microspheres in Sprague-Dawley (SD) rats. In addition, rhGH-PELA microspheres induced a greater response in total insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) than other rhGH formulations. With a hypophysectomized SD rat model, the pharmacological efficacy of rhGH-PELA microspheres was shown to be better than that from daily administration of rhGH solutions over 6 days based on body weight gain and width of the tibial growth plate. Histological examination of the injection sites indicated a significantly milder inflammatory response than that observed after injection of PLA and PLGA microspheres. Neither anti-rhGH antibodies nor the toxic effects on heart, liver and kidney were detectable after administration of rhGH-PELA microspheres in SD rats. These results suggest that rhGH-PELA microspheres have the potential to be clinically effective and safe when administered only once every two months, a dose regimen for better patient acceptance and compliance.
Xiong, Long; Zeng, Jianhua; Yao, Aihua; Tu, Qiquan; Li, Jingtang; Yan, Liang; Tang, Zhiming
2015-01-01
The regeneration of large bone defects is an osteoinductive, osteoconductive, and osteogenic process that often requires a bone graft for support. Limitations associated with naturally autogenic or allogenic bone grafts have demonstrated the need for synthetic substitutes. The present study investigates the feasibility of using novel hollow hydroxyapatite microspheres as an osteoconductive matrix and a carrier for controlled local delivery of bone morphogenetic protein 2 (BMP2), a potent osteogenic inducer of bone regeneration. Hollow hydroxyapatite microspheres (100±25 μm) with a core (60±18 μm) and a mesoporous shell (180±42 m2/g surface area) were prepared by a glass conversion technique and loaded with recombinant human BMP2 (1 μg/mg). There was a gentle burst release of BMP2 from microspheres into the surrounding phosphate-buffered saline in vitro within the initial 48 hours, and continued at a low rate for over 40 days. In comparison with hollow hydroxyapatite microspheres without BMP2 or soluble BMP2 without a carrier, BMP2-loaded hollow hydroxyapatite microspheres had a significantly enhanced capacity to reconstitute radial bone defects in rabbit, as shown by increased serum alkaline phosphatase; quick and complete new bone formation within 12 weeks; and great biomechanical flexural strength. These results indicate that BMP2-loaded hollow hydroxyapatite microspheres could be a potential new option for bone graft substitutes in bone regeneration. PMID:25609957
A virus-MIPs fluorescent sensor based on FRET for highly sensitive detection of JEV.
Liang, Caishuang; Wang, Huan; He, Kui; Chen, Chunyan; Chen, Xiaoming; Gong, Hang; Cai, Changqun
2016-11-01
Major stumbling blocks in the recognition and detection of virus are the unstable biological recognition element or the complex detection means. Here a fluorescent sensor based on virus-molecular imprinted polymers (virus-MIPs) was designed for specific recognition and highly sensitive detection of Japanese encephalitis virus (JEV). The virus-MIPs were anchored on the surface of silica microspheres modified by fluorescent dye, pyrene-1-carboxaldehyde (PC). The fluorescence intensity of PC can be enhanced by the principle of fluorescence resonance energy transfer (FRET), where virus acted as energy donor and PC acted as energy acceptor. The enhanced fluorescence intensity was proportional to the concentration of virus in the range of 24-960pM, with a limit of detection (LOD, 3σ) of 9.6pM, and the relative standard deviation was 1.99%. In additional, the specificity study confirmed the resultant MIPs has high-selectivity for JEV. This sensor would become a new key for the detection of virus because of its high sensitive, simple operation, high stability and low cost. Copyright © 2016. Published by Elsevier B.V.
Surface and capillary forces encountered by zinc sulfide microspheres in aqueous electrolyte.
Gillies, Graeme; Kappl, Michael; Butt, Hans-Jürgen
2005-06-21
The colloid probe technique was used to investigate the interactions between individual zinc sulfide (ZnS) microspheres and an air bubble in electrolyte solution. Incorporation of zinc ions into the electrolyte solution overcomes the disproportionate zinc ion dissolution and mimics high-volume-fraction conditions common in flotation. Determined interaction forces revealed a distinct lack of long-ranged hydrophobic forces, indicated by the presence of a DLVO repulsion prior to particle engulfment. Single microsphere contact angles were determined from particle-bubble interactions. Contact angles increased with decreasing radii and with surface oxidation. Surface modification by the absorption of copper and subsequently potassium O-ethyldithiocarbonate (KED) reduced repulsive forces and strongly increased contact angles.
Correlation of two bioadhesion assays: the everted sac technique and the CAHN microbalance.
Santos, C A; Jacob, J S; Hertzog, B A; Freedman, B D; Press, D L; Harnpicharnchai, P; Mathiowitz, E
1999-08-27
This contribution correlates two in vitro methods utilized to determine bioadhesion. One method, the everted intestinal sac technique, is a passive test for bioadhesion involving several polymer microspheres and a section of everted intestinal tissue. The other method, the CAHN microbalance, employs a CAHN dynamic contact angle analyzer with modified software to record the tensile forces measured as a single polymer microsphere is pulled from intestinal tissue. This study demonstrates that CAHN and everted sac experiments yield similar results when used to quantify the bioadhesive nature of polymer microsphere systems. A polymer showing high adhesion in one method also demonstrates high bioadhesion in the other method; polymers that exhibit high fracture strength and tensile work measurements with the CAHN microbalance also yield high binding percentages with the everted sac method. The polymers tested and reported here are poly(caprolactone) and different copolymer ratios of poly(fumaric-co-sebacic anhydride). The results of this correlation demonstrate that each method alone is a valuable indicator of bioadhesion.
On-demand drawing of high aspect-ratio, microsphere-tipped elastomeric micropillars
NASA Astrophysics Data System (ADS)
Li, Qiang; Kim, Jaeyoun
2017-08-01
High aspect-ratio elastomeric micropillars are widely used in a plethora of applications, such as functional surfaces, actuators, and sensors. Their fabrication at arbitrary positions on non-planar substrates, however, has rarely been reported. Here we demonstrate a new technique for facile fabrication of high aspect-ratio, microsphere-tipped elastomeric micropillars on structures with uncommon geometries. As a proof-of-concept exemplary application, a fiber optic contact sensor is realized by integrating a micropillar onto the end facet of an optical fiber. Overall, both the fabrication technique and the resulting outcomes of this work will add new tools to the toolbox of soft-MEMS and softrobotics.
A magnetic nanoscale Fe₃O₄/Pβ-CD composite as an efficient peroxidase mimetic for glucose detection.
Shi, Yun; Huang, Jun; Wang, Jiangning; Su, Ping; Yang, Yi
2015-10-01
Magnetic polymer particles with different surface functionalizations were prepared by a simple one-pot solvothermal method and characterized as peroxidase mimetics. The fluorescence enhancement obtained by attaching a β-cyclodextrin polymer (Pβ(-CD)) to the surfaces of Fe3O4 magnetic microspheres (Fe3O4 MMs) to generate a Fe3O4/P(β-CD) composite allowed the rapid, sensitive and selective analysis of glucose. The catalytic activity of the Fe3O4/P(β-CD) composite was evaluated with regard to the effects of catalyst particle size and species, pH value, level of catalyst, benzoic acid concentration and reaction time. Detection limits of 0.015 μM for H2O2 and 0.03 μM for glucose were determined when measuring at the 10-min mark. The presence of other saccharides, ion species, amino acids and proteins, had little effect on the results, and this technique was found to allow the analysis of glucose in human serum with high accuracy. The reusability of the Fe3O4/P(β-CD) composite was also investigated after 10 successive runs. A possible mechanism is proposed in which Fe3O4 plays a significant role in inducing fluorescence and P(β-CD) enhances the fluorescence signal, acting as both a stabilizer and a phase transfer agent. Copyright © 2015 Elsevier B.V. All rights reserved.
Imaging on a Shoestring: Cost-Effective Technologies for Probing Vadose Zone Transport Processes
NASA Astrophysics Data System (ADS)
Corkhill, C.; Bridge, J. W.; Barns, G.; Fraser, R.; Romero-Gonzalez, M.; Wilson, R.; Banwart, S.
2010-12-01
Key barriers to the widespread uptake of imaging technology for high spatial resolution monitoring of porous media systems are cost and accessibility. X-ray tomography, magnetic resonance imaging (MRI), gamma and neutron radiography require highly specialised equipment, controlled laboratory environments and/or access to large synchrotron facilities. Here we present results from visible light, fluorescence and autoradiographic imaging techniques developed at low cost and applied in standard analytical laboratories, adapted where necessary at minimal capital expense. UV-visible time lapse fluorescence imaging (UV-vis TLFI) in a transparent thin bed chamber enabled microspheres labelled with fluorescent dye and a conservative fluorophore solute (disodium fluorescein) to be measured simultaneously in saturated, partially-saturated and actively draining quartz sand to elucidate empirical values for colloid transport and deposition parameters distributed throughout the flow field, independently of theoretical approximations. Key results include the first experimental quantification of the effects of ionic strength and air-water interfacial area on colloid deposition above a capillary fringe, and the first direct observations of particle mobilisation and redeposition by moving saturation gradients during drainage. UV-vis imaging was also used to study biodegradation and reactive transport in a variety of saturated conditions, applying fluorescence as a probe for oxygen and nitrate concentration gradients, pH, solute transport parameters, reduction of uranium, and mapping of two-dimensional flow fields around a model dipole flow borehole system to validate numerical models. Costs are low: LED excitation sources (< US 50), flow chambers (US 200) and detectors (although a complete scientific-grade CCD set-up costs around US$ 8000, robust datasets can be obtained using a commercial digital SLR camera) mean that set-ups can be flexible to meet changing experimental requirements. The critical limitations of UV-vis fluorescence imaging are the need for reliable fluorescent probes suited to the experimental objective, and the reliance on thin-bed (2D) transparent porous media. Autoradiographic techniques address some of these limitations permit imaging of key biogeochemical processes in opaque media using radioactive probes, without the need for specialised radiation sources. We present initial calibration data for the use of autoradiography to monitor transport parameters for radionuclides (99-technetium), and a novel application of a radioactive salt tracer as a probe for pore water content, in model porous media systems.
NASA Astrophysics Data System (ADS)
Stevenson, M. E.; Blaschke, A. P.; Kirschner, A.
2010-12-01
Regulators need a dependable method that would enable them to calculate with confidence the setback distance of a drinking water well from a potential point of contamination. Since it is not permissible to perform field tests using pathogenic microorganisms, it is necessary to predict the transport of dangerous microbes in a different way, using surrogates. One such surrogate method involves using bacteriophages, which are viruses that are pathogenic to bacteria, but are not dangerous to humans. Another possible surrogate to model the potential travel time of microbial contamination is the use of synthetic microspheres; we will test microspheres ranging in size from 0.025 to 1 µm. The constraining factor for comparing the transport of microspheres and bacteriophages is the detection limit of the measuring apparatus. Appropriate measuring techniques are mandatory for a comparison. Traditionally, bacteriophages are measured using plaque forming analysis, the detection limit being one plaque forming unit per petri dish. In our study, the use of solid-phase cytometry for enumerating microspheres for wellhead protection projects is being investigated, as the detection limit using this technology is one cell per filter. To the best of our knowledge, there is no other technique available that enables a comparable detection limit. The solid-phase cytometer used for this study is a ChemScan RDI (Chemunex, France). For comparison, epifluorescence microscopy will also be used. The ChemScan RDI device automatically drives an epifluorescent microscope to the site of each cell detected, in order to confirm the validity of the reading. In this way, it is possible to observe whether clumping together of microspheres is a problem or if non-target cells were labelled. Keywords: Microspheres, Solid-phase cytometry, ChemScan, Drinking water protection Acknowledgements: We would like to thank the Austrian Science Fund (FWF) for financial support as part of the Doctoral Program on Water Resource Systems (DK Plus W1219-N22) and the Vienna Waterworks (MA 31) as part of the GWRS-Vienna project.
NASA Astrophysics Data System (ADS)
Mehdi, Yamina Ait; Itatahine, Asma; Fizir, Meriem; Xiao, Deli; Dramou, Pierre; He, Hua
2018-07-01
An ideal nanocarrier system for drug delivery is that one made from biocompatible and biodegradable materials for safe excretion from the biological system, and often with additional imaging abilities. In the present work, new core-shell silica microspheres have been prepared, with carrier decomposition after drug release. Paclitaxel, which is one of the most efficient drugs against a wide range of malignancies was integrated into the silica core. The carrier decomposition resulted from the escape of drug molecules with loading capacity about 16.95%. To achieve the fluorescents properties of the synthesized material a biocompatible photoluminescent prepared carbon dots were inserted in a silica shell around the Ptx-SiO2 core. The resultant silica core-shell (Ptx-SiO2CDs-SiO2) NPs with average particle size around 100 nm showed high fluorescent properties from the confocal laser scanning microscope observation. Further observation under UV-light at 365 nm also confirmed the photoluminescence. The Ptx-SiO2@CDs-SiO2 NPs were highly water soluble, and provide a sustained drug release as well as pH sensitivity. The incubation of A549 cells line with Ptx-SiO2@CDs-SiO2 NPs exhibits high cellular uptake as shown by CDs imaging. These properties in addition to the biocompatibility of Ptx-SiO2@CDs-SiO2 NPs and biodegradability of the silica core contributed simultaneously with the drug release process for easy body excretion after its functionality via renal system.
Analytical Applications of Fluorescent Carbon Dots
NASA Astrophysics Data System (ADS)
Goncalves, Helena Maria Rodrigues
The work described in this PhD Thesis focuses on the post-processing of optical fibers and their enhancement as sensing element. Since the majority of sensors presented are based in Fabry-Perot interferometers, an historical overview of this category of optical fiber sensors is firstly presented. This review considers the works published since the early years, in the beginning of the 1980s, until the middle of 2015. The incorporation of microcavities at the tip of a single mode fiber was extensively studied, particularly for the measurement of nitrogen and methane gas pressure. These cavities were fabricated using hollow core silica tubes and a hollow core photonic crystal fiber. Following a different approach, the microcavities were incorporated between two sections of single mode fiber. In this case, the low sensitivity to temperature makes these microcavities highly desirable for the measurement of strain at high temperatures. Competences in post-processing techniques such as the chemical etching and the writing of periodical structures in the fiber core by means of an excimer or a femtosecond laser were also acquired in the course of the PhD programme. One of the works consisted in the design and manufacturing of a double clad optical fiber. The refractive index of the inner cladding was higher than the one of the outer cladding and the core. Thus, light was guided in the inner cladding instead of propagating in the core. This situation was overcome by applying chemical etching, thus removing the inner cladding. The core, surrounded by air, was then able to guide light. Two different applications were found for this fiber, as a temperature sensor and as an optical refractometer. In the last, the optical phase changes with the liquid refractive index. Two different types of fiber Bragg gratings were characterized in strain and temperature. Sensing structures obtained through the phase mask technique at the tip of an optical fiber were subjected to chemical etching. In this case, an excimer laser was used. Extremely thin fiber tips were obtained, with an ultra-high sensitivity to strain. The other technique employed to fabricate the fiber Bragg gratings was the point-by-point femtosecond laser inscription. In this case, the sensing elements are very stable at high temperatures and can be used to measure strain in harsh conditions. The employment of optical fiber lasers as sensing elements was also considered in this Thesis. Two laser cavities were studied, one based on the ring configuration and the other based on a figure-of-eight configuration. From these works, the quality of the laser emission, namely the signal-to-noise ratio, the reduced full-width at half maximum and the stability should be highlighted. These characteristics allowed the measurement of different physical parameters, such as strain, temperature and torsion. Lastly, the possibility to use microspheres as sensing elements was considered. Using the electric arc of a fusion splicer, it is possible to create microspheres at the tip of an optical fiber. Furthermore, with this technique it is chains of microspheres can be obtained, constituting Mach-Zehnder-type interferometers which are sensitive to physical parameters like strain and temperature. The preliminary results obtained by introducing silica microspheres in a support structure are also presented. In this case, the sensors were subjected to temperature variations. All the experimental work was combined with the respective theoretical considerations. Many questions have been raised with the course of this PhD, and there are still some without a definite answer. Thus, new research paths can be followed, having their basis grounded in the configurations here presented.
El-Say, Khalid M; El-Helw, Abdel-Rahim M; Ahmed, Osama A A; Hosny, Khaled M; Ahmed, Tarek A; Kharshoum, Rasha M; Fahmy, Usama A; Alsawahli, Majed
2015-01-01
The purpose was to improve the encapsulation efficiency of cetirizine hydrochloride (CTZ) microspheres as a model for water soluble drugs and control its release by applying response surface methodology. A 3(3) Box-Behnken design was used to determine the effect of drug/polymer ratio (X1), surfactant concentration (X2) and stirring speed (X3), on the mean particle size (Y1), percentage encapsulation efficiency (Y2) and cumulative percent drug released for 12 h (Y3). Emulsion solvent evaporation (ESE) technique was applied utilizing Eudragit RS100 as coating polymer and span 80 as surfactant. All formulations were evaluated for micromeritic properties and morphologically characterized by scanning electron microscopy (SEM). The relative bioavailability of the optimized microspheres was compared with CTZ marketed product after oral administration on healthy human volunteers using a double blind, randomized, cross-over design. The results revealed that the mean particle sizes of the microspheres ranged from 62 to 348 µm and the efficiency of entrapment ranged from 36.3% to 70.1%. The optimized CTZ microspheres exhibited a slow and controlled release over 12 h. The pharmacokinetic data of optimized CTZ microspheres showed prolonged tmax, decreased Cmax and AUC0-∞ value of 3309 ± 211 ng h/ml indicating improved relative bioavailability by 169.4% compared with marketed tablets.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Hunt, R. D.; Collins, J. L.; Cowell, B. S.
Cerium dioxide (CeO 2) is a commonly used simulant for plutonium dioxide and for plutonium (Pu) in a mixed uranium (U) and Pu oxide [(U, Pu)O 2] in nuclear fuel development. This effort developed CeO 2 microspheres with different porosities and diameters for use in a crush-strength study. The internal gelation technique has produced CeO 2 microspheres with limited initial porosity. When an equal molar solution of urea and hexamethylenetetramine (HMTA) is gently boiling for 1 hr and used in the gelation process, the crystallite size and porosity of mixed U and thorium oxide microspheres and the (U, Pu)O 2more » microspheres increased significantly. In this study with cerium, the combination of ammonium cerium nitrate and 1-h boiled HMTA-urea failed to produce a stable feed broth. However, when the 1-h heated HMTA-urea was combined with unheated HMTA-urea in 1 to 3 volume ratio or the boiling time of the HMTA-urea was reduced to 15-20 min, a stable solution of HMTA, urea, and Ce was formed at 273 K. This new Ce solution produced CeO 2 microspheres with much higher initial porosities. Intermediate porosities were possible when the heated HMTA/urea was aged prior to use.« less
An Electrochemical DNA Microbiosensor Based on Succinimide-Modified Acrylic Microspheres
Ulianas, Alizar; Heng, Lee Yook; Hanifah, Sharina Abu; Ling, Tan Ling
2012-01-01
An electrochemical microbiosensor for DNA has been fabricated based on new acrylic microspheres modified with reactive N-acryloxysuccinimide (NAS) functional groups. Hydrophobic poly(n-butylacrylate-N-acryloxysuccinimide) microspheres were synthesized in an emulsion form with a simple one-step photopolymerization technique. Aminated DNA probe was attached to the succinimde functional group of the acrylic microspheres via covalent bonding. The hybridization of the immobilized DNA probe with the complementary DNA was studied by differential pulse voltametry using anthraquninone-2-sulfonic acid monohydrate sodium salt (AQMS) as the electroactive hybridization label. The influences of many factors such as duration of DNA probe immobilization and hybridization, pH, type of ions, buffer concentrations, ionic strength, operational temperature and non-complementary DNA on the biosensor performance were evaluated. Under optimized conditions, the DNA microbiosensor demonstrated a linear response range to target DNA over a wide concentration range of 1.0 × 10−16 and 1.0 × 10−8 M with a lower limit of detection (LOD) of 9.46 × 10−17 M (R2 = 0.97). This DNA microbiosensor showed good reproducibility with 2.84% RSD (relative standard deviation) (n = 3). Application of the NAS-modified acrylic microspheres in the construction of DNA microbiosensor had improved the overall analytical performance of the resultant DNA microbiosensor when compared with other reported DNA biosensors using other nano-materials for membranes and microspheres as DNA immobilization matrices. PMID:22778594
Hunt, R. D.; Collins, J. L.; Cowell, B. S.
2017-05-13
Cerium dioxide (CeO 2) is a commonly used simulant for plutonium dioxide and for plutonium (Pu) in a mixed uranium (U) and Pu oxide [(U, Pu)O 2] in nuclear fuel development. This effort developed CeO 2 microspheres with different porosities and diameters for use in a crush-strength study. The internal gelation technique has produced CeO 2 microspheres with limited initial porosity. When an equal molar solution of urea and hexamethylenetetramine (HMTA) is gently boiling for 1 hr and used in the gelation process, the crystallite size and porosity of mixed U and thorium oxide microspheres and the (U, Pu)O 2more » microspheres increased significantly. In this study with cerium, the combination of ammonium cerium nitrate and 1-h boiled HMTA-urea failed to produce a stable feed broth. However, when the 1-h heated HMTA-urea was combined with unheated HMTA-urea in 1 to 3 volume ratio or the boiling time of the HMTA-urea was reduced to 15-20 min, a stable solution of HMTA, urea, and Ce was formed at 273 K. This new Ce solution produced CeO 2 microspheres with much higher initial porosities. Intermediate porosities were possible when the heated HMTA/urea was aged prior to use.« less
Dhankar, Vandana; Garg, Garima; Dhamija, Koushal; Awasthi, Rajendra
2014-01-01
Mucoadhesion enables localization of drugs to a defined region of the gastrointestinal tract through attractive interactions between polymers composing the drug delivery devices and the mucin layer of the intestinal epithelium. Thus, this approach can be used for enhancement of the oral bioavailability of the drug. The current communication deals with the development of ranitidine hydrochloride-loaded chitosan-based mucoadhesive microspheres. Microspheres were prepared by water-in-oil emulsion technique, using glutaraldehyde as a cross-linking agent. The effect of independent variables like stirring speed and polymer-to-drug ratio on dependent variables, i.e. percentage mucoadhesion, percentage drug loading, particle size and swelling index, was examined using a 3(2); factorial design. The microspheres were discrete, spherical, free-flowing and also showed high percentage drug entrapment efficiency (43-70%). An in vitro mucoadhesion test showed that the microspheres adhered strongly to the mucous layer for an extended period of time. The RC 4 batch exhibited a high percentage of drug encapsulation (70%) and mucoadhesion (75%). The drug release was sustained for more than 12 h. The drug release kinetics were found to follow Peppas' kinetics for all the formulations and the drug release was diffusion controlled. The preliminary results of this study suggest that the developed microspheres containing ranitidine hydrochloride could enhance drug entrapment efficiency, reduce the initial burst release and modulate the drug release.
A microflow cytometer on a chip
NASA Astrophysics Data System (ADS)
Golden, Joel P.; Kim, Jason; Anderson, George P.; Hashemi, Nastaran; Howell, Peter J.; Ligler, Frances S.
2010-02-01
A rapid, automated, multi-analyte Microflow Cytometer is being developed as a portable, field-deployable sensor for onsite diagnosis of biothreat agent exposure and environmental monitoring. The technology relies on a unique method for ensheathing a sample stream in continuous flow past an interrogation region where optical fibers provide excitation and collect emission. This approach efficiently focuses particles in the interrogation region of the fluidic channel, avoids clogging and provides for subsequent separation of the core and sheath fluids in order to capture the target for confirmatory assays and recycling of the sheath fluid. Fluorescently coded microspheres provide the capability for highly multiplexed assays. Optical analysis at four different wavelengths identified six sets of the coded microspheres recognizing Escherichia coli, Listeria, and Salmonella as well as cholera toxin, staphylococcal enterotoxin B (SEB), and ricin, and assay results were compared with those of a commercial Luminex analysis system.
Synthesis of plastic scintillation microspheres: alpha/beta discrimination.
Santiago, L M; Bagán, H; Tarancón, A; Garcia, J F
2014-11-01
Plastic scintillation microspheres (PSm) have been developed as an alternative for liquid scintillation cocktails due to their ability to avoid the mixed waste, besides other strengths in which the possibility for alpha/beta discrimination is included. The aim of this work was to evaluate the capability of PSm containing two combinations of fluorescence solutes (PPO/POPOP and pT/Bis-MSB) and variable amounts of a second organic solvent (naphthalene) to enhance the alpha/beta discrimination. Two commercial detectors with different Pulse Shape Discrimination performances (Quantulus and Triathler) were used to evaluate the alpha/beta discrimination. An optimal discrimination of alpha/beta particles was reached, with very low misclassification values (2% for beta particles and 0.5% for alpha particles), when PSm containing PPO/POPOP and between 0.6 and 2.0 g of naphthalene were evaluated using Triathler and the appropriate programme for data processing. Copyright © 2014 Elsevier Ltd. All rights reserved.
NASA Astrophysics Data System (ADS)
Falzone, Tobias; Blair, Savanna; Robertson-Anderson, Rae
2015-03-01
The semiflexible biopolymer actin, a ubiquitous component of nearly all biological organisms, plays an important role in many mechanically-driven processes such as muscle contraction, cancer invasion and cell motility. As such, entangled actin networks, which possess unique and complex viscoelastic properties, have been the subject of much theoretical and experimental work. However, due to this viscoelastic complexity, much is still unknown regarding the correlation of the applied stress on actin networks to the induced filament strain at the molecular and micro scale. Here, we use simultaneous optical trapping and fluorescence microscopy to characterize the link between applied microscopic forces and strain propagation as a function of strain rate and concentration. Specifically, we track fiduciary markers on entangled actin filaments before, during and after actively driving embedded microspheres through the network. These measurements provide much needed insight into the molecular-level dynamics connecting stress and strain in semiflexible polymer networks.
Sharma, Neelam; Arora, Sandeep; Madan, Jitender
2018-02-01
Once-daily oral dosage of nefopam hydrochloride loaded sustained release microspheres (NPH-MS) was investigated as novel therapeutic strategy for post-operative pain management. Microspheres were synthesized using poly-3-hydroxybutyrate and poly-(ɛ-caprolactone) by double emulsion solvent evaporation technique. NPH-MS were characterized through FTIR, PXRD and SEM. In-vitro drug release study revealed sustained behavior till 24 h. Haemolysis was <5% which signified haemocompatibility of formulation. ED50 in rat tail-flick anti-nociceptive test was found ∼18.12 mg/kg. In post-operative pain model, reversal of mechanical allodynia and thermal hyperalgesia by NPH-MS was statistically significant (p < .001) as compared with NPH till 24 h post-dose.
Development of starch-gelatin complex microspheres as sustained release delivery system
Hari, B. N. Vedha; Praneetha, T.; Prathyusha, T.; Mounika, K.; Devi, D. Ramya
2012-01-01
The starch was isolated from jackfruit seeds and evaluated for its preformulation properties, like tapped density, bulk density, and particle size. The fourier transform infrared (FTIR) analysis was done and compared with that of the commercially available starch which confirmed the properties. Using the various concentrations of jackfruit seed starch, the microspheres were prepared, combining with gelatin by ionotropic gelation technique. The developed microspheres were subjected to analysis of particle size, drug content, entrapment efficiency, and percentage yield. The spectral analysis confirmed the presence of drug and absence of interactions. Scanning electron microscope image showed that the particles were in spherical shape with a rough surface. The in vitro drug release in water for 12 hours proved to be in the range of 89 to 100%. The various kinetic models were applied using release data to confirm the mechanism of drug. It was concluded that the jackfruit starch-gelatin microspheres gave satisfactory results and met pharmacopieal limits. PMID:23057005
de Azevedo, Mariangela de Burgos M; de Melo, Vitor H S; Soares, Carlos Roberto J; Miyamoto, Douglas M; Katayama, Ricardo A; Squair, Peterson L; Barros, Caio H N; Tasic, Ljubica
2018-06-14
Biodegradable polymers containing radioactive isotopes such as Holmium 166 ( 166 Ho) have potential applications as beta particle emitters in tumour tissues. It is also a gamma ray emitter, allowing nuclear imaging of any tissue to be acquired. It is frequently used in the form of complexes such as holmium acetylacetonate (HoAcAc), which may cause damages in tissues next to the targets cancer cells, as it is difficult to control its linkage or healthy tissues radiotherapy effects. Poly(d,l-lactic acid), PDLLA, was used to encapsulate holmium acetylacetonate (HoAcAc) using an emulsion solvent extraction/evaporation technique. Microspheres with sizes between 20-53 µm were extensively characterised. HoAcAc release from the microspheres was assessed through studies using Inductively Coupled Plasma - Optical Emission Spectroscopy, and the microspheres showed no holmium leakage after a period of 10 half-lives and following gamma irradiation. Thus, HoAcAc loaded microspheres are here presented as a potential system for brachytherapy and imaging purposes.
Chang, Fei; Xie, Yunchao; Chen, Juan; Luo, Jieru; Li, Chenlu; Hu, Xuefeng; Xu, Bin
2015-02-01
Preparation of uniform BiOCI flower-like microspheres was facilely accomplished through a sim- ple protocol involving regulation of pH value in aqueous with sodium hydroxide in the presence of n-propanol. The as-prepared samples were characterized by a collection of techniques, such as X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), UV-vis diffuse reflectance spectroscopy (UV-vis DRS), and nitrogen adsorption-desorption isotherms. Based upon the SEM analyses, uniform microspheres could be formed with coexistence of some fragments of BiOCI nanosheets without n-propanol. The addition of appropriate amount of n-propanol was beneficial to provide BiOCI samples containing only flower-like microspheres, which were further subjected to the photocatalytic measurements towards Rhodamine B in aqueous under visible light irradiation and exhibited the best catalytic performance among all samples tested. In addition, the photocatalytic process was confirmed to undergo through a photosensitization pathway, in which superoxide radicals (.O-) played critical roles.
Kelmendi-Doko, Arta; Rubin, J Peter; Klett, Katarina; Mahoney, Christopher; Wang, Sheri; Marra, Kacey G
2017-01-01
Current materials used for adipose tissue reconstruction have critical shortcomings such as suboptimal volume retention, donor-site morbidity, and poor biocompatibility. The aim of this study was to examine a controlled delivery system of dexamethasone to generate stable adipose tissue when mixed with disaggregated human fat in an athymic mouse model for 6 months. The hypothesis that the continued release of dexamethasone from polymeric microspheres would enhance both adipogenesis and angiogenesis more significantly when compared to the single-walled microsphere model, resulting in long-term adipose volume retention, was tested. Dexamethasone was encapsulated within single-walled poly(lactic-co-glycolic acid) microspheres (Dex SW MS) and compared to dexamethasone encapsulated in a poly(lactic-co-glycolic acid) core surrounded by a shell of poly-l-lactide. The double-walled polymer microsphere system in the second model was developed to create a more sustainable drug delivery process. Dexamethasone-loaded poly(lactic-co-glycolic acid) microspheres (Dex SW MS) and dexamethasone-loaded poly(lactic-co-glycolic acid)/poly-l-lactide double-walled microspheres (Dex DW MS) were prepared using single and double emulsion/solvent techniques. In vitro release kinetics were determined. Two doses of each type of microsphere were examined; 50 and 27 mg of Dex MS and Dex DW MS were mixed with 0.3 mL of human lipoaspirate. Additionally, 50 mg of empty MS and lipoaspirate-only controls were examined. Samples were analyzed grossly and histologically after 6 months in vivo. Mass and volume were measured; dexamethasone microsphere-containing samples demonstrated greater adipose tissue retention compared to the control group. Histological analysis, including hematoxylin and eosin and CD31 staining, indicated increased vascularization (p < 0.05) within the Dex MS-containing samples. Controlled delivery of adipogenic factors, such as dexamethasone via polymer microspheres, significantly affects adipose tissue retention by maintaining healthy tissue formation and vascularization. Dex DW MS provide an improved model to former Dex SW MS, resulting in notably longer release time and, consequently, larger volumes of adipose retained in vivo. The use of microspheres, specifically double-walled, as vehicles for controlled drug delivery of adipogenic factors therefore present a clinically relevant model of adipose retention that has the potential to greatly improve soft tissue repair. PMID:29051810
Holographic optical assembly and photopolymerized joining of planar microspheres
Shaw, L. A.; Chizari, S.; Panas, R. M.; ...
2016-07-27
The aim of this research is to demonstrate a holographically driven photopolymerization process for joining colloidal particles to create planar microstructures fixed to a substrate, which can be monitored with real-time measurement. Holographic optical tweezers (HOT) have been used to arrange arrays of microparticles prior to this work; here we introduce a new photopolymerization process for rapidly joining simultaneously handled microspheres in a plane. Additionally, we demonstrate a new process control technique for efficiently identifying when particles have been successfully joined by measuring a sufficient reduction in the particles’ Brownian motion. Furthermore, this technique and our demonstrated joining approach enablemore » HOT technology to take critical steps toward automated additive fabrication of microstructures.« less
NASA Astrophysics Data System (ADS)
Novakowski, K. S.; Trimper, S.; Praamsma, T.; Springthorpe, S.
2010-12-01
Shallow, unprotected bedrock aquifers are common sources of drinking water supply in eastern North America. The vulnerability of these aquifers to contamination from pathogens is widely recognised, although little is actually known about the transport processes involved, particularly where the source is located near to or on ground surface (i.e. a septic system). In this experiment we explore the transport of fluorescent microspheres having diameters of 1.75 and 0.3 µm and the bacteriophage Φ-X174 in a sparsely-fractured gneissic terrain having minimal overburden cover. The experiment was conducted by ponding water in a 7 m2 area on the edge of an outcrop having observable vertical fractures and measuring the arrival of particles in two nearby monitoring wells. A conservative solute tracer (Lissamine FF) was also used to follow the solute front. In order to encourage transport to the wells and to provide a discharge stream to sample, pumping was conducted at a rate of 7.7 L/min from the lower half of the 15-m deep well farthest from the pond (approximately 7 m away). Sampling was conducted from the pumping stream, the upper 5 m of that well and the upper 5 m of an additional well located about 5 m from the surface pond. The experiment was conducted over a 48 hr period and samples were obtained every 15 min initially declining to once every 2 hrs towards the end of the experiment. Analysis of the bacteriophage was conducted using the Double Agar Layer method and the concentration of microspheres was determined using epi-fluorescent microscopy. As the latter is very time consuming, only preliminary results are available for the microsphere transport. The results show widespread migration of both the microspheres and the bacteriophage, as arrival in all sampling locations was detected. Mass recovery was low but similar for both the bacteriophage and the solute tracer, although the majority of the bacteriophage arrived much earlier than the majority of the solute tracer particularly for the deeper sample. The degree of longitudinal dispersion experienced by the bacteriophage transport was also significantly less than that for the solute tracer.
Characterisation of the behaviour of particles in biofilters for pre-treatment of drinking water.
Persson, Frank; Långmark, Jonas; Heinicke, Gerald; Hedberg, Torsten; Tobiason, John; Stenström, Thor-Axel; Hermansson, Malte
2005-10-01
Biofiltration of surface water was examined using granular activated carbon (GAC) and expanded clay (EC). Particle removal was 60-90%, measured by flow cytometry, which enabled discrimination between total- and autofluorescent particles (microalgae) in size ranges of 0.4-1 and 1-15 microm, and measured by on-line particle counting. Total particles were removed at a higher degree than autofluorescent particles. The biofilters were also challenged with 1 microm fluorescent microspheres with hydrophobic and hydrophilic surface characteristics and bacteriophages (Salmonella typhimurium 28B). Added microspheres were removed at 97-99% (hydrophobic) and 85-89% (hydrophilic) after 5 hydraulic residence times (HRT) and microspheres retained in the biofilter media were slowly detaching into the filtrate for a long time after the addition. Removal of bacteriophages (5 HRT) was considerably lower at 40-59%, and no long-lasting detachment was observed. A comparison of experimental data with theoretical predictions for removal of particles in clean granular media filters revealed a similar or higher removal of particles around 1 microm in size than predicted, while bacteriophages were removed at a similar or lesser extent than predicted. The results highlight the selectivity and dynamic behaviour of the particle removal processes and have implications for operation and microbial risk assessment of a treatment train with biofilters as pre-treatment.
Xu, Chao; Wygladacz, Katarzyna; Retter, Robert; Bell, Michael; Bakker, Eric
2007-12-15
Polymeric bulk optode microsphere ion sensors in combination with suspension array technologies such as analytical flow cytometry may become a power tool for measuring electrolytes in physiological samples. In this work, the methodology for the direct measurement of common blood electrolytes in physiological samples using bulk optode microsphere sensors was explored. The simultaneous determination of Na(+), K(+), and Ca(2+) in diluted sheep blood plasma was demonstrated for the first time, using a random suspension array containing three types of mixed microsphere bulk optodes of similar size, fabricated from the same chromoionophore without additional labeling. Sodium ionophore X, potassium ionophore III, and grafted AU-1 in poly(butyl acrylate) were the ionophores used in the bulk optode microsphere ion sensors for Na(+), K(+), and Ca(2+), respectively, in combination with the cation-exchanger NaTFPB (sodium tetrakis-[3,5-bis(trifluoromethyl)phenyl]borate) and the same concentration of the chromoionophore ETH 5294 (9-(di-ethylamino)-5-octadecanoylimino-5H-benzo[a]phen-oxazine) in plasticized poly(vinyl chloride). Excellent reproducibility was achieved for the sensing of potassium ions. The effect of sample pH was relatively small at near-physiological pH and followed theoretical predictions, yet the sample temperature was found to influence the sensor response to a larger extent. Multiplexed ion sensing was achieved by taking advantage of the chemical tunability of the sensor response, adjusting the sensor compositions so that the three types of ion sensors responded with distinct levels of protonation of the chromoionophore. Consequently, three well-resolved peaks were simultaneously observed in the single-channel histogram during the multiplexed calibration as well as in the subsequent measurement of the three cations in 10-fold-diluted sheep plasma. The assigned peak positions corresponded very well to the physiological range of the measured ions.
Nonlinear optical effects in organic microstructures
NASA Astrophysics Data System (ADS)
Novikov, Vladimir B.; Mamonov, Evgeniy A.; Kopylov, Denis A.; Mitetelo, Nikolai V.; Venkatakrishnarao, D.; Narayana, YSLV; Chandrasekar, R.; Murzina, Tatiana V.
2017-05-01
Organic microstructures attract much attention due to their unique properties originating from the design of their shape and optical parameters. In this work we discuss the linear, second- and third-order nonlinear optical effects in arrays and in individual organic microstructures composed by self-assembling technique and formed randomly on top of a solid substrate. The structures under study consist of micro-spheres, -hemispheres or -frustums made of red laser dye and reveal an intense fluorescence (FL) in the visible spectral range. Importantly, that due to a high value of the refractive index and confined geometry, such micro-structures support the excitation of whispering gallery modes (WGM), which brings about strong and spectrally-selected light localization. We show that an amplification of the nonlinear optical effects is observed for these structures as compared to a homogeneous dye film of similar composition. The obtained data are in agreement with the results of the FDTD calculations performed for the structures of different dimensions. Perspectives of application of such type of organic nonlinear microresonators in optical devices are discussed.
Magnetically Actuated Cilia for Microfluidic Manipulation
NASA Astrophysics Data System (ADS)
Hanasoge, Srinivas; Owen, Drew; Ballard, Matt; Hesketh, Peter J.; Alexeev, Alexander; Woodruff School of Mechanical Engineering Collaboration; Petit InstituteBioengineering; Biosciences Collaboration
2015-11-01
We demonstrate magnetic micro-cilia based microfluidic mixing and capture techniques. For this, we use a simple and easy to fabricate high aspect ratio cilia, which are actuated magnetically. These micro-features are fabricated by evaporating NiFe alloy at room temperature, on to patterned photoresist. The evaporated alloy curls upwards when the seed layer is removed to release the cilia, thus making a free standing `C' shaped magnetic microstructure. This is actuated using an external electromagnet or a rotating magnet. The artificial cilia can be actuated upto 20Hz. We demonstrate the active mixing these cilia can produce in the microchannel. Also, we demonstrate the capture of target species in a sample using these fast oscillating cilia. The surface of the cilia is functionalized by streptavidin which binds to biotin labelled fluorescent microspheres and mimic the capture of bacteria. We show very high capture efficiencies by using these methods. These simple to fabricate micro cilia can easily be incorporated into many microfluidic systems which require high mixing and capture efficiencies.
Lucconi, Giulia; Chlapanidas, Theodora; Martino, Emanuela; Gaggeri, Raffaella; Perteghella, Sara; Rossi, Daniela; Faragò, Silvio; Vigo, Daniele; Faustini, Massimo; Collina, Simona; Torre, Maria Luisa
2014-02-01
Extracts of Crataegus monogyna Jacq. (hawthorn) show an interesting free radical scavenging (FRS) effect, related to their flavonoids content. Unfortunately, their oral administration is affected by their low bioavailability. The aim of this work is to obtain a multiparticulate drug delivery system for hawthorn extracts for oral administration. The extracts from flowering tops (FL) or fruits (FR) of hawthorn were obtained with maceration, using ethanol as an extraction solvent, and their antioxidant activity was evaluated. FL extract showed the highest FRS activity (EC50 3.72 ± 1.21 µg/ml), so it was selected to prepare microparticulate systems by a spray-drying technique, which were characterized by granulometric analysis, scanning electron microscopy-energy dispersive X-ray spectroscopy, confocal fluorescence microscopy and hyperoside content. Antioxidant activity was evaluated before and after gastrointestinal transit in vitro simulation. Results indicate that the microparticulate systems maintained the antioxidant activity of hawthorn also after gastrointestinal transit in vitro simulation, exhibiting properties suitable for oral administration.
Photonic nanojet super-resolution in immersed ordered assembly of dielectric microspheres
NASA Astrophysics Data System (ADS)
Geints, Y. E.; Zemlyanov, A. A.
2017-10-01
Specific spatially-localized optical field structure, which is often referred to as a photonic nanojet (PNJ), is formed in the near-field scattering area of non-absorbing dielectric micron-sized particle exposed to an optical radiation. By virtue of the finite-difference time-domain technique we numerically simulate the two-dimensional array of PNJs created by an ordered single-layer microassembly of glass microspheres immersed in a transparent polymer matrix. The behavior of the main PNJ parameters (length, diameter, and intensity) is analyzed subject to the immersion depth of the microparticles and cooperative interference effects of the neighboring microspheres. We show that depending on microassembly configuration, the PNJ quality can be significantly improved; in particular, the PNJ spatial resolution better than λ/5 can be achieved.
Long-term conversion of 45S5 bioactive glass-ceramic microspheres in aqueous phosphate solution.
Fu, Hailuo; Rahaman, Mohamed N; Day, Delbert E; Huang, Wenhai
2012-05-01
The conversion of 45S5 glass and glass-ceramics to a hydroxyapatite (HA)-like material in vitro has been studied extensively, but only for short reaction times (typically <3 months). In this paper, we report for the first time on the long-term conversion of 45S5 glass-ceramic microspheres (designated 45S5c) in an aqueous phosphate solution. Microspheres of 45S5c (75-150 μm) were immersed for 10 years at room temperature (~25 °C) in K(2)HPO(4) solution with a concentration of 0.01 M or 1.0 M, and with a starting pH of 7.0 or 9.5. The reacted 45S5c microspheres and solutions were analyzed using structural and analytical techniques. Only 25-45 vol% of the 45S5c microspheres were converted to an HA-like material after the 10 year reaction. In solutions with a starting pH of 9.5, an increase in the K(2)HPO(4) concentration from 0.01 to 1.0 M resulted in a doubling of the volume of the microspheres converted to an HA-like material but had little effect on the composition of the HA-like product. In comparison, reaction of the 45S5c microspheres in the solution with a starting pH of 7.0 resulted in an HA-like product in the 0.01 M K(2)HPO(4) solution but a calcium pyrophosphate product, Ca(10)K(4)(P(2)O(7))(6).9H(2)O, in the 1.0 M solution. The consequences of these results for the long-term use of 45S5 glass-ceramics in biomedical applications are discussed.
Processing and Characterization of Sol-Gel Cerium Oxide Microspheres
DOE Office of Scientific and Technical Information (OSTI.GOV)
McClure, Zachary D.; Padilla Cintron, Cristina
Of interest to space exploration and power generation, Radioisotope Thermoelectric Generators (RTGs) can provide long-term power to remote electronic systems without the need for refueling or replacement. Plutonium-238 (Pu-238) remains one of the more promising materials for thermoelectric power generation due to its high power density, long half-life, and low gamma emissions. Traditional methods for processing Pu-238 include ball milling irregular precipitated powders before pressing and sintering into a dense pellet. The resulting submicron particulates of Pu-238 quickly accumulate and contaminate glove boxes. An alternative and dust-free method for Pu-238 processing is internal gelation via sol-gel techniques. Sol-gel methodology createsmore » monodisperse and uniform microspheres that can be packed and pressed into a pellet. For this study cerium oxide microspheres were produced as a surrogate to Pu-238. The similar electronic orbitals between cerium and plutonium make cerium an ideal choice for non-radioactive work. Before the microspheres can be sintered and pressed they must be washed to remove the processing oil and any unreacted substituents. An investigation was performed on the washing step to find an appropriate wash solution that reduced waste and flammable risk. Cerium oxide microspheres were processed, washed, and characterized to determine the effectiveness of the new wash solution.« less
Liu, Dongfei; Chen, Jian; Jiang, Tao; Li, Wei; Huang, Yao; Lu, Xiyi; Liu, Zehua; Zhang, Weixia; Zhou, Zheng; Ding, Qirui; Santos, Hélder A; Yin, Guoyong; Fan, Jin
2018-04-01
New treatment strategies for spinal cord injury with good therapeutic efficacy are actively pursued. Here, acetalated dextran (AcDX), a biodegradable polymer obtained by modifying vicinal diols of dextran, is demonstrated to protect the traumatically injured spinal cord. To facilitate its administration, AcDX is formulated into microspheres (≈7.2 µm in diameter) by the droplet microfluidic technique. Intrathecally injected AcDX microspheres effectively reduce the traumatic lesion volume and inflammatory response in the injured spinal cord, protect the spinal cord neurons from apoptosis, and ultimately, recover the locomotor function of injured rats. The neuroprotective feature of AcDX microspheres is achieved by sequestering glutamate and calcium ions in cerebrospinal fluid. The scavenging of glutamate and calcium ion reduces the influx of calcium ions into neurons and inhibits the formation of reactive oxygen species. Consequently, AcDX microspheres attenuate the expression of proapoptotic proteins, Calpain, and Bax, and enhance the expression of antiapoptotic protein Bcl-2. Overall, AcDX microspheres protect traumatically injured spinal cord by alleviating the glutamate-induced excitotoxicity. This study opens an exciting perspective toward the application of neuroprotective AcDX for the treatment of severe neurological diseases. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Song, Kedong; Liu, Yingchao; Macedo, Hugo M; Jiang, Lili; Li, Chao; Mei, Guanyu; Liu, Tianqing
2013-04-01
Nutrient depletion within three-dimensional (3D) scaffolds is one of the major hurdles in the use of this technology to grow cells for applications in tissue engineering. In order to help in addressing it, we herein propose to use the controlled release of encapsulated nutrients within polymer microspheres into chitosan-based 3D scaffolds, wherein the microspheres are embedded. This method has allowed maintaining a stable concentration of nutrients within the scaffolds over the long term. The polymer microspheres were prepared using multiple emulsions (w/o/w), in which bovine serum albumin (BSA) and poly (lactic-co-glycolic) acid (PLGA) were regarded as the protein pattern and the exoperidium material, respectively. These were then mixed with a chitosan solution in order to form the scaffolds by cryo-desiccation. The release of BSA, entrapped within the embedded microspheres, was monitored with time using a BCA kit. The morphology and structure of the PLGA microspheres containing BSA before and after embedding within the scaffold were observed under a scanning electron microscope (SEM). These had a round shape with diameters in the range of 27-55 μm, whereas the chitosan-based scaffolds had a uniform porous structure with the microspheres uniformly dispersed within their 3D structure and without any morphological change. In addition, the porosity, water absorption and degradation rate at 37 °C in an aqueous environment of 1% chitosan-based scaffolds were (92.99±2.51) %, (89.66±0.66) % and (73.77±3.21) %, respectively. The studies of BSA release from the embedded microspheres have shown a sustained and cumulative tendency with little initial burst, with (20.24±0.83) % of the initial amount released after 168 h (an average rate of 0.12%/h). The protein concentration within the chitosan-based scaffolds after 168 h was found to be (11.44±1.81)×10(-2) mg/mL. This novel chitosan-based scaffold embedded with PLGA microspheres has proven to be a promising technique for the development of new and improved tissue engineering scaffolds. Copyright © 2012 Elsevier B.V. All rights reserved.
Development of Yersinia pestis F1 antigen-loaded microspheres vaccine against plague
Huang, Shih-shiung; Li, I-Hsun; Hong, Po-da; Yeh, Ming-kung
2014-01-01
Yersinia pestis F1 antigen-loaded poly(DL-lactide-co-glycolide)/polyethylene glycol (PEG) (PLGA/PEG) microspheres were produced using a water-in-oil-in-water emulsion/solvent extraction technique and assayed for their percent yield, entrapment efficiency, surface morphology, particle size, zeta potential, in vitro release properties, and in vivo animal protect efficacy. The Y. pestis F1 antigen-loaded microspheres (mean particle size 3.8 μm) exhibited a high loading capacity (4.5% w/w), yield (85.2%), and entrapment efficiency (38.1%), and presented a controlled in vitro release profile with a low initial burst (18.5%), then continued to release Y. pestis F1 antigen over 70 days. The distribution (%) of Y. pestis F1 on the microspheres surface, outer layer, and core was 3.1%, 28.9%, and 60.7%, respectively. A steady release rate was noticed to be 0.55 μg Y. pestis F1 antigen/mg microspheres/day of Y. pestis F1 antigen release maintained for 42 days. The cumulative release amount at the 1st, 28th, and 42nd days was 8.2, 26.7, and 31.0 μg Y. pestis F1 antigen/mg microspheres, respectively. The 100 times median lethal dose 50% (LD50) of Y. pestis Yokohama-R strain by intraperitoneal injection challenge in mice test, in which mice received one dose of 40 μg F1 antigen content of PLGA/PEG microspheres, F1 antigen in Al(OH)3, and in comparison with F1 antigen in Al(OH)3 vaccine in two doses, was evaluated after given by subcutaneous immunization of BALB/c mice. The study results show that the greatest survival was observed in the group of mice immunized with one dose of F1 antigen-loaded PLGA/PEG microspheres, and two doses of F1 antigen in Al(OH)3 vaccine (100%). In vivo vaccination studies also demonstrated that F1 vaccines microspheres had a protective ability; its steady-state IgG immune protection in mice plasma dramatic increased from 2 weeks (18,764±3,124) to 7 weeks (126,468±19,176) after vaccination. These findings strongly suggest that F1-antigen loaded microspheres vaccine offer a new therapeutic strategy in optimizing the vaccine incorporation and delivery properties of these potential vaccine targeting carriers. PMID:24550673
Yang, Fuhua; Yu, Fan; Zhang, Zhian; Zhang, Kai; Lai, Yanqing; Li, Jie
2016-02-12
Sodium-ion batteries (SIBs) are regarded as an attractive alternative to lithium-ion batteries (LIBs) for large-scale commercial applications, because of the abundant terrestrial reserves of sodium. Exporting suitable anode materials is the key to the development of SIBs and LIBs. In this contribution, we report on the fabrication of Bi@C microspheres using aerosol spray pyrolysis technique. When used as SIBs anode materials, the Bi@C microsphere delivered a high capacity of 123.5 mAh g(-1) after 100 cycles at 100 mA g(-1) . The rate performance is also impressive (specific capacities of 299, 252, 192, 141, and 90 mAh g(-1) are obtained under current densities of 0.1, 0.2, 0.5, 1, and 2 A g(-1) , respectively). Furthermore, the Bi@C microsphere also proved to be suitable LIB anode materials. The excellent electrochemical performance for both SIBs and LIBs can attributed to the Bi@C microsphere structure with Bi nanoparticles uniformly dispersed in carbon spheres. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Acosta, Niuris; Sánchez, Elisa; Calderón, Laura; Cordoba-Diaz, Manuel; Cordoba-Diaz, Damián; Dom, Senne; Heras, Ángeles
2015-01-01
A chitosan-based hydrophilic system containing an olive leaf extract was designed and its antioxidant capacity was evaluated. Encapsulation of olive leaf extract in chitosan microspheres was carried out by a spray-drying process. The particles obtained with this technique were found to be spherical and had a positive surface charge, which is an indicator of mucoadhesiveness. FTIR and X-ray diffraction results showed that there are not specific interactions of polyphenolic compounds in olive leaf extract with the chitosan matrix. Stability and release studies of chitosan microspheres loaded with olive leaf extract before and after the incorporation into a moisturizer base were performed. The resulting data showed that the developed formulations were stable up to three months. The encapsulation efficiency was around 44% and the release properties of polyphenols from the microspheres were found to be pH dependent. At pH 7.4, polyphenols release was complete after 6 h; whereas the amount of polyphenols released was 40% after the same time at pH 5.5. PMID:26389926
DOE Office of Scientific and Technical Information (OSTI.GOV)
Melo, Ronaldo P. de; Colégio Militar do Recife, Exército Brasileiro, Recife PE 50730-120; Oliveira, Nathalia Talita C.
A novel procedure based on a two-step method was developed to obtain β-Ga{sub 2}O{sub 3} nanowires by the chemical vapor deposition (CVD) method. The first step consists in the gallium micro-spheres growth inside a metal-organic chemical vapor deposition environment, using an organometallic precursor. Nanoscale spheres covering the microspheres were obtained. The second step involves the CVD oxidization of the gallium micro-spheres, which allow the formation of β-Ga{sub 2}O{sub 3} nanowires on the micro-sphere surface, with the final result being a nanostructure mimicking nature's sea urchin morphology. The grown nanomaterial is characterized by several techniques, including X-ray diffraction, scanning electron microscopy,more » energy-dispersive X-ray, transmission electron microscopy, and photoluminescence. A discussion about the growth mechanism and the optical properties of the β-Ga{sub 2}O{sub 3} material is presented considering its unknown true bandgap value (extending from 4.4 to 5.68 eV). As an application, the scattering properties of the nanomaterial are exploited to demonstrate random laser emission (around 570 nm) when it is permeated with a laser dye liquid solution.« less
Qu, Jian-Bo; Chen, Yan-Li; Huan, Guan-Sheng; Zhou, Wei-Qing; Liu, Jian-Guo; Zhu, Hu; Zhang, Xiao-Yun
2015-01-01
A high-speed thermoresponsive medium was developed by grafting poly(N-isopropylacrylamide-co-butyl methacrylate) (P(NIPAM-co-BMA)) brushes onto gigaporous polystyrene (PS) microspheres via surface-initiated atom transfer radical polymerization (ATRP) technique, which has strong mechanical strength, good chemical stability and high mass transfer rate for biomacromolecules. The gigaporous structure, surface chemical composition, static protein adsorption, and thermoresponsive chromatographic properties of prepared medium (PS-P(NIPAM-co-BMA)) were characterized in detail. Results showed that the PS microspheres were successfully grafted with P(NIPAM-co-BMA) brushes and that the gigaporous structure was robustly maintained. After grafting, the nonspecific adsorption of proteins on PS microspheres was greatly reduced. A column packed with PS-P(NIPAM-co-BMA) exhibited low backpressure and significant thermo-responsibility. By simply changing the column temperature, it was able to separate three model proteins at the mobile phase velocity up to 2167 cm h(-1). In conclusion, the thermoresponsive polymer brushes grafted gigaporous PS microspheres prepared by ATRP are very promising in 'green' high-speed preparative protein chromatography. Copyright © 2014 Elsevier B.V. All rights reserved.
Cao, Ruofan; Naivar, Mark A; Wilder, Mark; Houston, Jessica P
2014-01-01
Fluorescence lifetime measurements provide information about the fluorescence relaxation, or intensity decay, of organic fluorophores, fluorescent proteins, and other inorganic molecules that fluoresce. The fluorescence lifetime is emerging in flow cytometry and is helpful in a variety of multiparametric, single cell measurements because it is not impacted by nonlinearity that can occur with fluorescence intensity measurements. Yet time-resolved cytometry systems rely on major hardware modifications making the methodology difficult to reproduce. The motivation of this work is, by taking advantage of the dynamic nature of flow cytometry sample detection and applying digital signal processing methods, to measure fluorescence lifetimes using an unmodified flow cytometer. We collect a new lifetime-dependent parameter, referred to herein as the fluorescence-pulse-delay (FPD), and prove it is a valid representation of the average fluorescence lifetime. To verify we generated cytometric pulses in simulation, with light emitting diode (LED) pulsation, and with true fluorescence measurements of cells and microspheres. Each pulse is digitized and used in algorithms to extract an average fluorescence lifetime inherent in the signal. A range of fluorescence lifetimes is measurable with this approach including standard organic fluorophore lifetimes (∼1 to 22 ns) as well as small, simulated shifts (0.1 ns) under standard conditions (reported herein). This contribution demonstrates how digital data acquisition and signal processing can reveal time-dependent information foreshadowing the exploitation of full waveform analysis for quantification of similar photo-physical events within single cells. © 2014 The Authors. Published by Wiley Periodicals, Inc. PMID:25274073
NASA Astrophysics Data System (ADS)
Burtsev, Vasilii; Marchuk, Valentina; Kugaevskiy, Artem; Guselnikova, Olga; Elashnikov, Roman; Miliutina, Elena; Postnikov, Pavel; Svorcik, Vaclav; Lyutakov, Oleksiy
2018-03-01
Nano-spheres lithography is actually considered as a powerful tool to manufacture various periodic structures with a wide potential in the field of nano- and micro-fabrication. However, during self-assembling of colloid microspheres, various defects and mismatches can appear. In this work the size and quality of single-domains of closed-packed polystyrene (PS), grown up on thin Au layers modified by hydrophilic or hydrophobic functional groups via diazonium chemistry was studied. The effects of the surface modification on the quality and single-domain size of polystyrene (PS) microspheres array were investigated and discussed. Modified surfaces were characterized using the AFM and wettability tests. PS colloidal suspension was deposited using the drop evaporation method. Resulted PS microspheres array was characterized using the SEM, AFM and confocal microscopy technique.
NASA Astrophysics Data System (ADS)
Mbanjwa, Mesuli B.; Chen, Hao; Fourie, Louis; Ngwenya, Sibusiso; Land, Kevin
2014-06-01
Multiplexed or parallelised droplet microfluidic systems allow for increased throughput in the production of emulsions and microparticles, while maintaining a small footprint and utilising minimal ancillary equipment. The current paper demonstrates the design and fabrication of a multiplexed microfluidic system for producing biocatalytic microspheres. The microfluidic system consists of an array of 10 parallel microfluidic circuits, for simultaneous operation to demonstrate increased production throughput. The flow distribution was achieved using a principle of reservoirs supplying individual microfluidic circuits. The microfluidic devices were fabricated in poly (dimethylsiloxane) (PDMS) using soft lithography techniques. The consistency of the flow distribution was determined by measuring the size variations of the microspheres produced. The coefficient of variation of the particles was determined to be 9%, an indication of consistent particle formation and good flow distribution between the 10 microfluidic circuits.
Twala, Busisiwe V; Sewell, B Trevor; Jordaan, Justin
2012-05-10
The use of enzymes in industrial applications is limited by their instability, cost and difficulty in their recovery and re-use. Immobilisation is a technique which has been shown to alleviate these limitations in biocatalysis. Here we describe the immobilisation of two biocatalytically relevant co-factor recycling enzymes, glucose dehydrogenase (GDH) and NADH oxidase (NOD) on aldehyde functional ReSyn™ polymer microspheres with varying functional group densities. The successful immobilisation of the enzymes on this new high capacity microsphere technology resulted in the maintenance of activity of ∼40% for GDH and a maximum of 15.4% for NOD. The microsphere variant with highest functional group density of ∼3500 μmol g⁻¹ displayed the highest specific activity for the immobilisation of both enzymes at 33.22 U mg⁻¹ and 6.75 U mg⁻¹ for GDH and NOD with respective loading capacities of 51% (0.51 mg mg⁻¹) and 129% (1.29 mg mg⁻¹). The immobilised GDH further displayed improved activity in the acidic pH range. Both enzymes displayed improved pH and thermal stability with the most pronounced thermal stability for GDH displayed on ReSyn™ A during temperature incubation at 65 °C with a 13.59 fold increase, and NOD with a 2.25-fold improvement at 45 °C on the same microsphere variant. An important finding is the suitability of the microspheres for stabilisation of the multimeric protein GDH. Copyright © 2012 Elsevier Inc. All rights reserved.
Shi, Feng; Liu, Yumei; Zhi, Wei; Xiao, Dongqin; Li, Hongyu; Duan, Ke; Qu, Shuxin; Weng, Jie
2017-06-06
Microstructure and chemical constitution are important factors affecting the biological activity of biomaterials. This study aimed to fabricate hydroxyapatite (HAp) particles with both micro/nanohybrid structure and Cu 2+ doping to promote osteogenic differentiation and antibacterial property. In the presence of inositol hexakisphosphate (IP6), micro/nano-structured and Cu 2+ -doped HAp (HAp-IP6-Cu) microspheres were successfully fabricated via hydrothermal method. Morphological observation showed that HAp-IP6-Cu microspheres with a diameter of 3.1-4.1 μm were chrysanthemum-like and composed of nano-flakes approximately 50 nm in thickness. Compared with the HAp micro-rods or IP6 modified HAp (HAp-IP6) microspheres, HAp-IP6-Cu microspheres had a larger specific surface area, better hydrophilicity and stronger ability to adsorb bovine serum albumin. To evaluate the synergistic effects of micro/nanohybrid structure and Cu 2+ on cell behavior, rat calvarial osteoblasts (RCOs) were cultured on HAp-IP6-Cu, HAp-IP6 and HAp layers as well as their extracts, respectively. Results demonstrated that HAp-IP6-Cu layer promoted the adhesion, proliferation and osteogenic differentiation of RCOs. The cells grew on HAp-IP6-Cu and HAp-IP6 layers exhibited greater spreading than those on HAp layer. In addition, quantitative test by the agar disk diffusion technique found that HAp-IP6-Cu microspheres were effectively against S taphylococcus aureus and E scherichia coli. These results demonstrated that HAp-IP6-Cu microspheres may be a potential candidate as a bioactive and anti-infective biomaterial for bone regeneration.
PLGA microspheres encapsulating siRNA.
De Rosa, Giuseppe; Salzano, Giuseppina
2015-01-01
The therapeutic use of small interfering RNA (siRNA) represents a new and powerful approach to suppress the expression of pathologically genes. However, biopharmaceutical drawbacks, such as short half-life, poor cellular uptake, and unspecific distribution into the body, hamper the development of siRNA-based therapeutics. Poly(lactide-co-glycolide), (PLGA) microspheres can be a useful tool to overcome these issues. siRNA can be encapsulated into the PLGA microspheres, which protects the loaded nucleic acid against the enzymatic degradation. Moreover, PLGA microspheres can be injected directly into the action site, where the siRNA can be released in controlled manner, thus avoiding the need of frequent invasive administrations. The complete biodegradability of PLGA to monomers easily metabolized by the body, and its approval by FDA and EMA for parenteral administration, assure the safety of this copolymer and do not require the removal of the device after the complete drug release. In chapter, a basic protocol for the preparation of PLGA microspheres encapsulating siRNA is described. This protocol is based on a double emulsion/solvent evaporation technique, a well known and easy to reproduce method. This specific protocol has been developed to encapsulate a siRNA anti-TNFα in PLGA microspheres, and it has been designed and optimized to achieve high siRNA encapsulation efficiency and slow siRNA release in vitro. However, it can be extended also to other siRNA as well as other RNA or DNA-based oligonucleotides (miRNA, antisense, decoy, etc.). Depending on the applications, chemical modifications of the backbone and site-specific modification within the siRNA sequences could be required.
End-specific strategies of attachment of long double stranded DNA onto gold-coated nanofiber arrays
NASA Astrophysics Data System (ADS)
Peckys, Diana B.; de Jonge, Niels; Simpson, Michael L.; McKnight, Timothy E.
2008-10-01
We report the effective and site-specific binding of long double stranded (ds)DNA to high aspect ratio carbon nanofiber arrays. The carbon nanofibers were first coated with a thin gold layer to provide anchorage for two controllable binding methods. One method was based on the direct binding of thiol end-labeled dsDNA. The second and enhanced method used amine end-labeled dsDNA bound with crosslinkers to a carboxyl-terminated self-assembled monolayer. The bound dsDNA was first visualized with a fluorescent, dsDNA-intercalating dye. The specific binding onto the carbon nanofiber was verified by a high resolution detection method using scanning electron microscopy in combination with the binding of neutravidin-coated fluorescent microspheres to the immobilized and biotinylated dsDNA. Functional activity of thiol end-labeled dsDNA on gold-coated nanofiber arrays was verified with a transcriptional assay, whereby Chinese hamster lung cells (V79) were impaled upon the DNA-modified nanofibers and scored for transgene expression of the tethered template. Thiol end-labeled dsDNA demonstrated significantly higher expression levels than nanofibers prepared with control dsDNA that lacked a gold-binding end-label. Employing these site-specific and robust techniques of immobilization of dsDNA onto nanodevices can be of advantage for the study of DNA/protein interactions and for gene delivery applications.
Scanning superlens microscopy for non-invasive large field-of-view visible light nanoscale imaging
NASA Astrophysics Data System (ADS)
Wang, Feifei; Liu, Lianqing; Yu, Haibo; Wen, Yangdong; Yu, Peng; Liu, Zhu; Wang, Yuechao; Li, Wen Jung
2016-12-01
Nanoscale correlation of structural information acquisition with specific-molecule identification provides new insight for studying rare subcellular events. To achieve this correlation, scanning electron microscopy has been combined with super-resolution fluorescent microscopy, despite its destructivity when acquiring biological structure information. Here we propose time-efficient non-invasive microsphere-based scanning superlens microscopy that enables the large-area observation of live-cell morphology or sub-membrane structures with sub-diffraction-limited resolution and is demonstrated by observing biological and non-biological objects. This microscopy operates in both non-invasive and contact modes with ~200 times the acquisition efficiency of atomic force microscopy, which is achieved by replacing the point of an atomic force microscope tip with an imaging area of microspheres and stitching the areas recorded during scanning, enabling sub-diffraction-limited resolution. Our method marks a possible path to non-invasive cell imaging and simultaneous tracking of specific molecules with nanoscale resolution, facilitating the study of subcellular events over a total cell period.
Henning, Paul E.; Rigo, M. Veronica; Geissinger, Peter
2012-01-01
A highly porous optical-fiber cladding was developed for evanescent-wave fiber sensors, which contains sensor molecules, maintains guiding conditions in the optical fiber, and is suitable for sensing in aqueous environments. To make the cladding material (a poly(ethylene) glycol diacrylate (PEGDA) polymer) highly porous, a microsphere templating strategy was employed. The resulting pore network increases transport of the target analyte to the sensor molecules located in the cladding, which improves the sensor response time. This was demonstrated using fluorescein-based pH sensor molecules, which were covalently attached to the cladding material. Scanning electron microscopy was used to examine the structure of the templated polymer and the large network of interconnected pores. Fluorescence measurements showed a tenfold improvement in the response time for the templated polymer and a reliable pH response over a pH range of five to nine with an estimated accuracy of 0.08 pH units. PMID:22654644
NASA Astrophysics Data System (ADS)
Selvi, Bilge
In an effort to design novel formulation strategies to optimize the antitumor activity of camptothecin (CPT), the physicochemical and membrane binding properties of the drug, were investigated by various techniques in acidic and physiological pH. The intrinsic solubility of the CPT-lactone free base was determined to be 3.44 muM and 5.11 muM at 22°C and 37°C, respectively. The equilibrium solubility of the drug was found to increase with increasing temperature and decreasing pH. The enhanced solubility of the drug at very low pH is attributed to the protonation of the nitrogen atom in the ring B and the increased solvency of the highly acidic media. The logarithmic value of the intrinsic partition coefficient P of the free base CPT-lactone form was estimated to be 1.65, characteristic of a molecule suitable for oral absorption. The association constants Kf of the drug for bilayers composed of the zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and the negatively-charged 1,2-dioleoyl-sn-glycero-3-phospho- rac-(1-glycerol) (DOPG) were studied at acidic pH by fluorescence anisotropy and determined to be 35.4 +/- 4.5 M-1 and 93.1 +/- 11.0 M-1 for DOPC and DOPG, respectively, indicating a tendency of CPT to preferentially bind to negatively charged membranes. The energy of activation for the hydrolysis of CPT at physiological pH was found to be 114.3 +/- 33.4 kj/mole. The calculated t½ of the reaction at pH 7.2 at temperatures 25°C and 10°C was found to be 0.07 days and 5.12 days, respectively, whereas the time required for 1% of CPT-lactone to hydrolyze to CPT-carboxylate (t99%) was determined to be 1.8 hours, thus offering enough time to safely handle CPT-lactone at low temperatures. The preformulation results indicated that at highly acidic media CPT is positively charged and exists at its stable lactone form of increased solubility and has a capacity to bind to negatively charged membranes. Taking advantage of the increased stability of CPT in acidic media CPT-loaded microspheres were prepared in a 10 N HCl-methylene chloride mixture using the H-series of poly(D,L-lactide-co-glycolide) (H-PLGA). The system was then compared with a standard microsphere formation method and the results were evaluated with respect to particle morphology and drug release profile. Rough surface of the particles were obtained from the preparation method where a 10 N HCl solution was used. The release pattern of CPT was biphasic comprising a first burst effect followed by zero order release for all the formulations. However, the release of the drug was slightly faster from the microspheres formed with the modified method compared to the standard. Until now clinical application of CPT has been highly restricted by the insolubility and instability of the drug in its active lactone form, resulting in less antitumor potency and poor bioavailability. The pH-dependent release of the CPT-loaded microspheres was also compared and faster initial release (burst phase) was found at neutral pH, whereas at low pH the release was zero order for all the formulations. The results indicate that the stabilization and sustained release of CPT from H-PLGA microspheres might reduce local toxicity while simultaneously prolonging efficiency, suggesting new perspectives in CPT chemotherapy.
Leng, Yuankui
2017-01-01
Spectrometrically or optically encoded microsphere based suspension array technology (SAT) is applicable to the high-throughput, simultaneous detection of multiple analytes within a small, single sample volume. Thanks to the rapid development of nanotechnology, tremendous progress has been made in the multiplexed detecting capability, sensitivity, and photostability of suspension arrays. In this review, we first focus on the current stock of nanoparticle-based barcodes as well as the manufacturing technologies required for their production. We then move on to discuss all existing barcode-based bioanalysis patterns, including the various labels used in suspension arrays, label-free platforms, signal amplification methods, and fluorescence resonance energy transfer (FRET)-based platforms. We then introduce automatic platforms for suspension arrays that use superparamagnetic nanoparticle-based microspheres. Finally, we summarize the current challenges and their proposed solutions, which are centered on improving encoding capacities, alternative probe possibilities, nonspecificity suppression, directional immobilization, and “point of care” platforms. Throughout this review, we aim to provide a comprehensive guide for the design of suspension arrays, with the goal of improving their performance in areas such as multiplexing capacity, throughput, sensitivity, and cost effectiveness. We hope that our summary on the state-of-the-art development of these arrays, our commentary on future challenges, and some proposed avenues for further advances will help drive the development of suspension array technology and its related fields. PMID:26021602
Chen, T-H; Wang, S-Y; Chen, K-N; Liu, J-R; Chen, M-J
2009-07-01
In this study, various yeasts (Kluyveromyces marxianus, Saccharomyces turicensis, Pichia fermentans) and lactic acid bacteria (Lactobacillus kefiranofaciens, Lactobacillus kefiri, Leuconostoc mesenteroides) were entrapped in 2 different microspheres using an entrapment ratio for the strains that was based on the distribution ratio of these organisms in kefir grains. The purpose of this study was to develop a new technique to produce kefir using immobilized starter cultures isolated from kefir grains. An increase in cell counts with fermentation cycles was observed for both the lactic acid bacteria (LAB) and yeasts, whereas the cell counts of kefir grains were very stable during cultivation. Scanning electron microscopy showed that the short-chain lactobacilli and lactococci occupied the surface of the LAB microspheres, whereas the long-chain lactobacilli were inside the microspheres. When the yeasts were analyzed, cells at a high density were entrapped in cracks on the surface and within the microspheres, where they were surrounded by the short-chain lactobacilli. The distribution of the LAB and yeast species in kefir produced from grains and microspheres showed that there was no significant difference between the kefirs produced by the 2 methods; moreover, Leu. mesenteroides and K. marxianus were the predominating microflora in both types of kefir. There was no significant difference in the ethanol and exopolysaccharide contents between the 2 kefirs, although the acidity was different.
Lai, Shenzhi; Ouyang, Xiaoli; Cai, Changqun; Xu, Wensheng; Chen, Chunyan; Chen, Xiaoming
2017-05-01
The surface imprinting technique has been developed to overcome the mass-transfer difficulty, but the utilization ratio of template molecules in the imprinting procedure still remains a challengeable task to be improved. In this work, specifically designed surface-imprinted microspheres were prepared by a template-oriented method for enantioseparation of amlodipine besylate. Submicron mesoporous silica microspheres were surface-modified with double bonds, followed by polymerizing methacrylic acid to generate carboxyl modified mesoporous silica microspheres (PMAA@SiO 2 ). Afterwards, PMAA@SiO 2 was densely adsorbed with (S)-amlodipine molecules to immobilize template molecules through multiple hydrogen bonding interactions. Then surface molecular imprinting was carried out by cross-linking the carboxyl group of PMAA@SiO 2 with ethylene glycol diglycidyl ether. The surface-imprinted microspheres showed fast binding kinetics of only 20 min for equilibrium adsorption, and the saturation adsorption capacity reached 137 mg/g. The imprinted materials displayed appreciable chiral separation ability when used as column chromatography for enantioseparation of amlodipine from amlodipine besylate, and the enantiomeric excess of (S)-amlodipine reached 13.8% with only 2.3 cm column length by no extra chiral additives. Besides, the imprinted materials exhibited excellent reusability, and this allows the potential application for amplification production of amlodipine enantiomer. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Caisso, Marie; Picart, Sébastien; Belin, Renaud C; Lebreton, Florent; Martin, Philippe M; Dardenne, Kathy; Rothe, Jörg; Neuville, Daniel R; Delahaye, Thibaud; Ayral, André
2015-04-14
Transmutation of americium in heterogeneous mode through the use of U1-xAmxO2±δ ceramic pellets, also known as Americium Bearing Blankets (AmBB), has become a major research axis. Nevertheless, in order to consider future large-scale deployment, the processes involved in AmBB fabrication have to minimize fine particle dissemination, due to the presence of americium, which considerably increases the risk of contamination. New synthesis routes avoiding the use of pulverulent precursors are thus currently under development, such as the Calcined Resin Microsphere Pelletization (CRMP) process. It is based on the use of weak-acid resin (WAR) microspheres as precursors, loaded with actinide cations. After two specific calcinations under controlled atmospheres, resin microspheres are converted into oxide microspheres composed of a monophasic U1-xAmxO2±δ phase. Understanding the different mechanisms during thermal conversion, that lead to the release of organic matter and the formation of a solid solution, appear essential. By combining in situ techniques such as XRD and XAS, it has become possible to identify the key temperatures for oxide formation, and the corresponding oxidation states taken by uranium and americium during mineralization. This paper thus presents the first results on the mineralization of (U,Am) loaded resin microspheres into a solid solution, through in situ XAS analysis correlated with HT-XRD.
Nanofibrous spongy microspheres enhance odontogenic differentiation of human dental pulp stem cells.
Kuang, Rong; Zhang, Zhanpeng; Jin, Xiaobing; Hu, Jiang; Gupte, Melanie J; Ni, Longxing; Ma, Peter X
2015-09-16
Dentin regeneration is challenging due to its complicated anatomical structure and the shortage of odontoblasts. In this study, a novel injectable cell carrier, nanofibrous spongy microspheres (NF-SMS), is developed for dentin regeneration. Biodegradable and biocompatible poly(l-lactic acid)-block-poly(l-lysine) are synthesized and fabricated into NF-SMS using self-assembly and thermally induced phase separation techniques. It is hypothesized that NF-SMS with interconnected pores and nanofibers can enhance the proliferation and odontogenic differentiation of human dental pulp stem cells (hDPSCs), compared to nanofibrous microspheres (NF-MS) without pore structure and conventional solid microspheres (S-MS) with neither nanofibers nor pore structure. During the first 9 d in culture, hDPSCs proliferate significantly faster on NF-SMS than on NF-MS or S-MS (p < 0.05). Following in vitro odontogenic induction, all the examined odontogenic genes (alkaline phosphatase content, osteocalcin, bone sialoprotein, collagen 1, dentin sialophosphoprotein (DSPP)), calcium content, and DSPP protein content are found significantly higher in the NF-SMS group than in the control groups. Furthermore, 6 weeks after subcutaneous injection of hDPSCs and microspheres into nude mice, histological analysis shows that NF-SMS support superior dentin-like tissue formation compared to NF-MS or S-MS. Taken together, NF-SMS have great potential as an injectable cell carrier for dentin regeneration. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Boni, Fernanda Isadora; Prezotti, Fabíola Garavello; Cury, Beatriz Stringhetti Ferreira
2016-08-01
Gellan gum microspheres were obtained by ionotropic gelation technique, using the trivalent ion Al(3+). The percentage of entrapment efficiency ranged from 48.76 to 87.52% and 2(2) randomized full factorial design demonstrated that both the increase of polymer concentration and the decrease of crosslinker concentration presented a positive effect in the amount of encapsulated drug. Microspheres size and circularity ranged from 700.17 to 938.32 μm and from 0.641 to 0.796 μm, respectively. The increase of polymer concentration (1-2%) and crosslinker concentration (3-5%) led to the enlargement of particle size and circularity. However, the association of increased crosslinker concentration and reduced polymer content made the particles more irregular. In vitro and ex vivo tests evidenced the high mucoadhesiveness of microspheres. The high liquid uptake ability of the microspheres was demonstrated and the pH variation did not affect this parameter. Drug release was pH dependent, with low release rates in acid pH (42.40% and 44.93%) and a burst effect in phosphate buffer pH (7.4). The Weibull model had the best correlation with the drug release data, demonstrating that the release process was driven by a complex mechanism involving the erosion and swelling of the matrix or by non-Fickian diffusion.
Synthesis and Characterization of LaTiO2N
NASA Astrophysics Data System (ADS)
Rugen, Evan E.
Photocatalysts offer an excellent opportunity to shift the global energy landscape from a fossil fuel-dependent paradigm to sustainable and carbon-neutral solar fuels. Oxynitride materials such as LaTiO2N are potential photocatalysts for the water splitting reaction due to their high oxidative stability and their narrow band gaps, which are suitable for visible light absorption. However, facile synthetic routes to metal oxynitrides with controlled morphologies are rare, and the local structures of these materials are under-characterized. Ultrasonic spray synthesis (USS) offers a facile method toward complex metal oxides which can potentially be converted to oxynitrides with preservation of the microsphere structures that typify the products from such aerosol routes. Here, La-Ti-O microspheres were facilely produced by USS and converted by ammonolysis to LaTiO2N microspheres with porous shells and hollow interiors. This particle architecture is accounted for by coupling suitable combustion chemistry with the aerosol technique, producing precursor particles where the La3+ and Ti4+ are well-mixed at small length scales; this feature enables preservation of the microsphere morphology during nitridation despite the crystallographic changes that occur. The LaTiO2N microspheres are comparable oxygen evolving photocatalysts to samples produced by conventional solid state methods. Pair distribution function (PDF) analysis is a local probe designed to examine the structure of disordered crystalline materials, and is an ideal technique for characterizing the ordering of anions in oxynitrides. Preliminary studies using PDF analysis to determine the presence of anion ordering and local structure in LaTiO2N produced by solid state methods are presented here. Future experiments are proposed that will grant detailed insight into the factors driving the degree of anion ordering in these types of materials. These results demonstrate the utility of USS as a facile, potentially scalable route to complex photocatalytic materials and their precursors, and the feasibility of PDF analysis for the determination of local structures in complex oxynitrides.
Instrument for fluorescence sensing of circulating cells with diffuse light in mice in vivo.
Zettergren, Eric; Vickers, Dwayne; Runnels, Judith; Murthy, Shashi K; Lin, Charles P; Niedre, Mark
2012-03-01
Accurate quantification of circulating cell populations in mice is important in many areas of preclinical biomedical research. Normally, this is done either by extraction and analysis of small blood samples or, more recently, by using microscopy-based in vivo fluorescence flow cytometry. We describe a new technological approach to this problem using detection of diffuse fluorescent light from relatively large blood vessels in vivo. The diffuse fluorescence flow cytometer (DFFC) uses a laser to illuminate a mouse limb and an array of optical fibers coupled to a high-sensitivity photomultiplier tube array operating in photon counting mode to detect weak fluorescence signals from cells. We first demonstrate that the DFFC instrument is capable of detecting fluorescent microspheres and Vybrant-DiD-labeled cells in a custom-made optical flow phantom with similar size, optical properties, linear flow rates, and autofluorescence as a mouse limb. We also present preliminary data demonstrating that the DFFC is capable of detecting circulating cells in nude mice in vivo. In principle, this device would allow interrogation of the whole blood volume of a mouse in minutes, with sensitivity improvement by several orders of magnitude compared to current approaches. © 2012 Society of Photo-Optical Instrumentation Engineers (SPIE).
Site-specific multipoint fluorescence measurement system with end-capped optical fibers.
Song, Woosub; Moon, Sucbei; Lee, Byoung-Cheol; Park, Chul-Seung; Kim, Dug Young; Kwon, Hyuk Sang
2011-07-10
We present the development and implementation of a spatially and spectrally resolved multipoint fluorescence correlation spectroscopy (FCS) system utilizing multiple end-capped optical fibers and an inexpensive laser source. Specially prepared end-capped optical fibers placed in an image plane were used to both collect fluorescence signals from the sample and to deliver signals to the detectors. The placement of independently selected optical fibers on the image plane was done by monitoring the end-capped fiber tips at the focus using a CCD, and fluorescence from specific positions of a sample were collected by an end-capped fiber, which could accurately represent light intensities or spectral data without incurring any disturbance. A fast multipoint spectroscopy system with a time resolution of ∼1.5 ms was then implemented using a prism and an electron multiplying charge coupled device with a pixel binning for the region of interest. The accuracy of our proposed system was subsequently confirmed by experimental results, based on an FCS analysis of microspheres in distilled water. We expect that the proposed multipoint site-specific fluorescence measurement system can be used as an inexpensive fluorescence measurement tool to study many intracellular and molecular dynamics in cell biology. © 2011 Optical Society of America
Metaxa, Aikaterini-Foteini; Efthimiadou, Eleni K; Boukos, Nikos; Fragogeorgi, Eirini A; Loudos, George; Kordas, George
2014-12-01
Conventional chemotherapy drugs such as anthracyclines show no specific activity. They destroy cancer cells but also and the healthy ones, and for that reason exhibit high toxicity. In order to alleviate the toxic effects of chemotherapeutic drugs, the administration dose is being minimized, while their reactivity against tumor cells is lessened. This problem can be overcome or at least reduced by using nanoscale drug delivery systems to target the pathogenic area. The present work deals with the synthesis, characterization and biological evaluation of multi-responsive hollow microspheres coated with Hydroxypropyl Cellulose (HPC)-a biocompatible and thermosensitive polysaccharide-conjugated with folic acid as well promising drug vehicles for targeted cancer therapy. The synthetic route consists of two steps. In the first step, a single layer of sensitive copolymers is ((Methacrylic acid (MAA), N-(2-Hydroxypropyl) methacrylamide (HPMA) and N,N'-(disulfanediylbis(ethane-2,1-diyl))bis(2-methylacrylamide) (DSBMA)) fabricated on a sacrificial template of SiO2 and in the second step, an additional layer of the folic acid modified HPC coat the microspheres' surface. The layers fabrication is performed through a combination of distillation precipitation co-polymerization and chemical deposition method. The loading capacity (% LC) and encapsulation efficiency (% EE) percentages of the chemotherapeutic agent daunorubicin (DNR) in the fabricated microspheres were calculated through the standard curve methodology. In addition, the releasing properties of the resulting spheres are investigated, using the above mentioned methodology. It is worth mentioning that, spheres release the entrapped drug under combined conditions such acidic and reductive environment along with conventional hyperthermia. Cytotoxic activity of the synthesized spheres was investigated by using the well-established method of MTT assay in MCF-7 (breast cancer), HeLa (cervical cancer) and HEK 293 (Human Embryonic Kidney healthy cells) cell lines. Confocal and fluorescence microscopy were used to confirm the in vitro targeted ability of folic acid modified drug loaded microspheres in HeLa, to that overexpress folate receptors, MCF-7 and 3T3 cells, as negative folate cell substrate. Finally, radiolabelling of the spheres is performed, with a gamma emitting radionuclide ((99m)Tc), to assess their in vivo profile by means of scintigraphic imaging and biodistribution studies. Hollow spheres release the encapsulated drug under acidic environment, conventional hyperthermia or in the presence of glutathione (reductive environment). The ability of modified drug carriers to target the HeLa cells, was confirmed by confocal and fluorescence microscopy. The resulting spheres are observed to be promising drug-carriers for cancer treatment due to their releasing properties under tumor's environment and high concentration in HeLa cells via endocytosis. In addition, the empty vehicles have no toxicity in healthy cells and present antimicrobial activity. Copyright © 2014 Elsevier Inc. All rights reserved.
Braaf, Boy; de Boer, Johannes F
2017-03-20
Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.
"Smart tattoo" glucose biosensors and effect of coencapsulated anti-inflammatory agents.
Srivastava, Rohit; Jayant, Rahul Dev; Chaudhary, Ayesha; McShane, Michael J
2011-01-01
Minimally invasive glucose biosensors with increased functional longevity form one of the most promising techniques for continuous glucose monitoring. In the present study, we developed a novel nanoengineered microsphere formulation comprising alginate microsphere glucose sensors and anti-inflammatory-drug-loaded alginate microspheres. The formulation was prepared and characterized for size, shape, in vitro drug release, biocompatibility, and in vivo acceptability. Glucose oxidase (GOx)- and Apo-GOx-based glucose sensors were prepared and characterized. Sensing was performed both in distilled water and simulated interstitial body fluid. Layer-by-layer self-assembly techniques were used for preventing drug and sensing chemistry release. Finally, in vivo studies, involving histopathologic examination of subcutaneous tissue surrounding the implanted sensors using Sprague-Dawley rats, were performed to test the suppression of inflammation and fibrosis associated with glucose sensor implantation. The drug formulation showed 100% drug release with in 30 days with zero-order release kinetics. The GOx-based sensors showed good enzyme retention and enzyme activity over a period of 1 month. Apo-GOx-based visible and near-infrared sensors showed good sensitivity and analytical response range of 0-50 mM glucose, with linear range up to 12 mM glucose concentration. In vitro cell line studies proved biocompatibility of the material used. Finally, both anti-inflammatory drugs were successful in controlling the implant-tissue interface by suppressing inflammation at the implant site. The incorporation of anti-inflammatory drug with glucose biosensors shows promise in improving sensor biocompatibility, thereby suggesting potential application of alginate microspheres as "smart tattoo" glucose sensors with increased functional longevity. © 2010 Diabetes Technology Society.
Checa-Casalengua, Patricia; Jiang, Caihui; Bravo-Osuna, Irene; Tucker, Budd A; Molina-Martínez, Irene T; Young, Michael J; Herrero-Vanrell, Rocío
2011-11-30
The present experimental work describes the use of a novel protein encapsulation method to achieve protection of the biological factor during the microencapsulation procedure. With this aim, the protein is included in poly(lactic-co-glycolic acid) (PLGA) microspheres without any preliminary manipulation, in contrast to the traditional S/O/W (solid-in-oil-in-water) method where the bioactive substance is first dissolved and then freeze-dried in the presence of lyoprotectors. Furthermore, the presented technique involves the use of an oily additive, vitamin E (Vit E), useful from a technological point of view, by promoting additional protein protection and also from a pharmacological point of view, because of its antioxidant and antiproliferative properties. Application of this microencapsulation technique has been performed for GDNF (glial cell line-derived neurotrophic factor) designed for the treatment of optic nerve degenerative diseases, such as glaucoma, the second leading cause of blindness in the western world. The protein was released in vitro in its bioactive form for more than three months, demonstrated by the survival of their potential target cells (photoreceptors and retinal ganglion cells (RGC)). Moreover, the intravitreal injection of GDNF/Vit E PLGA microspheres in an experimental animal model of glaucoma significantly increased RGC survival compared with GDNF, Vit E or blank microspheres (p<0.01). This effect was present for at least eleven weeks, which suggests that the formulation prepared may be clinically useful as a neuroprotective tool in the treatment of glaucomatous optic neuropathy. Copyright © 2011 Elsevier B.V. All rights reserved.
NASA Astrophysics Data System (ADS)
Rout, Dipak; Kumar, Govind; Vijaya, R.
2018-01-01
A photonic crystal hetero-structure consisting of a passive planar defect of SiO2 thin film sandwiched between two identical opals grown by inward growing self-assembly method using Rhodamine-B dye-doped polystyrene microspheres is studied for the characteristics of dye emission. The optical properties and the defect mode characteristics of the hetero-structure are studied from the reflection and transmission measurements. Laser-induced fluorescence from the hetero-structure showed amplified and spectrally narrowed emission compared to the photonic crystal emphasizing the role of the defect mode and distributed feedback. The enhanced emission is also complemented by the reduction in fluorescence decay time in the case of the hetero-structure in comparison to the 3D photonic crystals.
Bittner, B; Witt, C; Mäder, K; Kissel, T
1999-08-05
The aim of the present study was to investigate the influence of the chemical insertion of poly(ethylene oxide), PEO, into a poly(lactide-co-glycolide), PLG, backbone on the mechanisms of in vitro degradation and erosion of the polymer. For this purpose microspheres prepared by a modified W/O/W double emulsion technique using ABA triblock copolymers, consisting of PLG A-blocks attached to central PEO B-blocks were compared with microspheres prepared from PLG. Due to their molecular architecture the ABA triblock copolymers differed in their erosion and degradation behavior from PLG. Degradation occurred faster in the ABA polymers by cleavage of ester bonds inside the polymer backbone. Even erosion was shown to start immediately after incubation in different buffer media. By varying pH and ionic strength of the buffer it was found that both mass loss and molecular weight decay were accelerated in alkaline and acidic pH in the case of the ABA triblock copolymers. Although the pH of the medium had a moderate influence on the degradation of PLG, the molecular weight decay was not accompanied by a mass loss during the observation time. In a second set of experiments we prepared bovine serum albumin, BSA, loaded microspheres from both polymers. The release of BSA from ABA microspheres under in vitro conditions parallels the faster swelling and erosion rates. This could be confirmed by electron paramagnetic resonance, EPR, measurements with spin labeled albumin where an influx of buffer medium into the ABA microspheres was already observed within a few minutes. In contrast, PLG microspheres revealed a burst release without any erosion. The current study shows that the environmental conditions affected the degradation and erosion of the pure polymer microspheres in the same way as the release of the model protein. This leads to the conclusion that the more favorable degradation profile of the ABA triblock copolymers was responsible for the improvement of the release profile.
Simultaneous Determination of Glass Transition Temperatures of Several Polymers.
He, Jiang; Liu, Wei; Huang, Yao-Xiong
2016-01-01
A simple and easy optical method is proposed for the determination of glass transition temperature (Tg) of polymers. Tg was determined using the technique of microsphere imaging to monitor the variation of the refractive index of polymer microsphere as a function of temperature. It was demonstrated that the method can eliminate most thermal lag and has sensitivity about six fold higher than the conventional method in Tg determination. So the determined Tg is more accurate and varies less with cooling/heating rate than that obtained by conventional methods. The most attractive character of the method is that it can simultaneously determine the Tg of several polymers in a single experiment, so it can greatly save experimental time and heating energy. The method is not only applicable for polymer microspheres, but also for the materials with arbitrary shapes. Therefore, it is expected to be broadly applied to different fundamental researches and practical applications of polymers.
Single-beam dielectric-microsphere trapping with optical heterodyne detection
NASA Astrophysics Data System (ADS)
Rider, Alexander D.; Blakemore, Charles P.; Gratta, Giorgio; Moore, David C.
2018-01-01
A technique to levitate and measure the three-dimensional position of micrometer-sized dielectric spheres with heterodyne detection is presented. The two radial degrees of freedom are measured by interfering light transmitted through the microsphere with a reference wavefront, while the axial degree of freedom is measured from the phase of the light reflected from the surface of the microsphere. This method pairs the simplicity and accessibility of single-beam optical traps to a measurement of displacement that is intrinsically calibrated by the wavelength of the trapping light and has exceptional immunity to stray light. A theoretical shot noise limit of 1.3 ×10-13 m /√{Hz } for the radial degrees of freedom, and 3.0 ×10-15 m /√{Hz } for the axial degree of freedom can be obtained in the system described. The measured acceleration noise in the radial direction is 7.5 ×10-5 (m /s2) /√{Hz } .
Molybdenum-base cermet fuel development
NASA Astrophysics Data System (ADS)
Pilger, James P.; Gurwell, William E.; Moss, Ronald W.; White, George D.; Seifert, David A.
Development of a multimegawatt (MMW) space nuclear power system requires identification and resolution of several technical feasibility issues before selecting one or more promising system concepts. Demonstration of reactor fuel fabrication technology is required for cermet-fueled reactor concepts. The MMW reactor fuel development activity at Pacific Northwest Laboratory (PNL) is focused on producing a molybdenum-matrix uranium-nitride (UN) fueled cermte. This cermet is to have a high matrix density (greater than or equal to 95 percent) for high strength and high thermal conductance coupled with a high particle (UN) porosity (approximately 25 percent) for retention of released fission gas at high burnup. Fabrication process development involves the use of porous TiN microspheres as surrogate fuel material until porous Un microspheres become available. Process development was conducted in the areas of microsphere synthesis, particle sealing/coating, and high-energy-rate forming (HERF) and the vacuum hot press consolidation techniques. This paper summarizes the status of these activities.
Yu, Meiling; Yao, Qing; Zhang, Yan; Chen, Huilin; He, Haibing; Zhang, Yu; Yin, Tian; Tang, Xing; Xu, Hui
2018-02-27
Highly soluble drugs tend to release from preparations at high speeds, which make them need to be taken at frequent intervals. Additionally, some drugs need to be controlled to release in vivo at certain periods, so as to achieve therapeutic effects. Thus, the objective of this study is to design injectable microparticulate systems with controllable in vivo release profile. Biodegradable PLGA was used as the matrix material to fabricate microspheres using the traditional double emulsification-solvent evaporation method as well as improved techniques, with gel (5% gelatine or 25% F127) or LP powders as the inner phases. Their physicochemical properties were systemically investigated. Microspheres prepared by modified methods had an increase in drug loading (15.50, 16.72, 15.66%, respectively) and encapsulation efficiencies (73.46, 79.42, 74.40%, respectively) when compared with traditional methods (12.01 and 57.06%). The morphology of the particles was characterized by optical microscope (OM) and scanning electron microscopy (SEM), and the amorphous nature of the encapsulated drug was confirmed by differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis. To evaluate their release behaviour, the in vitro degradation, in vitro release and in vivo pharmacodynamics were subsequently studied. Traditional microspheres prepared in this study with water as the inner phase had a relatively short release period within 16 d when compared with modified microspheres with 5% gelatine as the inner phase, which resulted in a smooth release profile and appropriate plasma LP concentrations over 21 d. Thus this type of modified microspheres can be better used in drugs requiring sustained release. The other two formulations containing 25% F127 and LP micropowders presented two-stage release profiles, resulting in fluctuant plasma LP concentrations which may be suitable for drugs requiring controlled release. All the results suggested that drug release rates from the microspheres prepared by various methods were mainly controlled by either the porosity inside the microspheres or the degradation of materials, which could, therefore, lead to different release behaviours. This results indicated great potential of the PLGA microsphere formulation as an injectable depot for controllable in vivo release profile via rational core phase design. Core/shell microspheres fabricated by modified double emulsification-solvent evaporation methods, with various inner phases, to obtain high loading drugs system, as well as appropriate release behaviours. Accordingly, control in vivo release profile via rational core phase design.
Scaffolds for bone regeneration made of hydroxyapatite microspheres in a collagen matrix.
Cholas, Rahmatullah; Kunjalukkal Padmanabhan, Sanosh; Gervaso, Francesca; Udayan, Gayatri; Monaco, Graziana; Sannino, Alessandro; Licciulli, Antonio
2016-06-01
Biomimetic scaffolds with a structural and chemical composition similar to native bone tissue may be promising for bone tissue regeneration. In the present work hydroxyapatite mesoporous microspheres (mHA) were incorporated into collagen scaffolds containing an ordered interconnected macroporosity. The mHA were obtained by spray drying of a nano hydroxyapatite slurry prepared by the precipitation technique. X-ray diffraction (XRD) analysis revealed that the microspheres were composed only of hydroxyapatite (HA) phase, and energy-dispersive x-ray spectroscopy (EDS) analysis revealed the Ca/P ratio to be 1.69 which is near the value for pure HA. The obtained microspheres had an average diameter of 6 μm, a specific surface area of 40 m(2)/g as measured by Brunauer-Emmett-Teller (BET) analysis, and Barrett-Joyner-Halenda (BJH) analysis showed a mesoporous structure with an average pore diameter of 16 nm. Collagen/HA-microsphere (Col/mHA) composite scaffolds were prepared by freeze-drying followed by dehydrothermal crosslinking. SEM observations of Col/mHA scaffolds revealed HA microspheres embedded within a porous collagen matrix with a pore size ranging from a few microns up to 200 μm, which was also confirmed by histological staining of sections of paraffin embedded scaffolds. The compressive modulus of the composite scaffold at low and high strain values was 1.7 and 2.8 times, respectively, that of pure collagen scaffolds. Cell proliferation measured by the MTT assay showed more than a 3-fold increase in cell number within the scaffolds after 15 days of culture for both pure collagen scaffolds and Col/mHA composite scaffolds. Attractive properties of this composite scaffold include the potential to load the microspheres for drug delivery and the controllability of the pore structure at various length scales. Copyright © 2016 Elsevier B.V. All rights reserved.
Silva, Camilla V N S; Barbosa, Jéssica A P; Ferraz, Milena S; Silva, Nicácio H; Honda, Neli K; Rabello, Marcelo M; Hernandes, Marcelo Z; Bezerra, Beatriz P; Cavalcanti, Isabella M F; Ayala, Alejandro P; Santos, Noemia P S; Santos-Magalhães, Nereide S
2016-11-01
In this pioneer study, 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) was used to improve the solubility of the diffractaic acid (DA) via inclusion complex (DA:HP-β-CD). Subsequently, DA:HP-β-CD was incorporated into poly-ε-caprolactone (PCL) microspheres (DA:HP-β-CD-MS). Microspheres containing DA (DA-MS) or DA:HP-β-CD (DA:HP-β-CD-MS) were prepared using the multiple W/O/W emulsion-solvent evaporation technique. The phase-solubility diagram of DA in HP-β-CD (10-50mM) showed an A L type curve with a stability constant K 1:1 =821M -1 . 1 H NMR, FTIR, X-ray diffraction and thermal analysis showed changes in the molecular environment of DA in DA:HP-β-CD. The molecular modeling approach suggests a guest-host complex formation between the carboxylic moiety of both DA and the host (HP-β-CD). The mean particle size of the microspheres were ∅ DA-MS =5.23±1.65μm and ∅ DA:HP-β-CD-MS =4.11±1.39μm, respectively. The zeta potential values of the microspheres were ζ DA-MS =-7.85±0.32mV and ζ DA:HP-β-CD-MS =-6.93±0.46mV. Moreover, the encapsulation of DA:HP-β-CD into microspheres resulted in a more slower release (k 2 =0.042±0.001; r 2 =0.996) when compared with DA-MS (k 2 =0.183±0.005; r 2 =0.996). The encapsulation of DA or DA:HP-β-CD into microspheres reduced the cytotoxicity of DA (IC 50 =43.29μM) against Vero cells (IC 50 of DA-MS=108.48μM and IC 50 of DA:HP-β-CD-MS=142.63μM). Copyright © 2016 Elsevier B.V. All rights reserved.
Cephradin-plaga microspheres for sustained delivery to cattle.
Ustariz-Peyret, C; Coudane, J; Vert, M; Kaltsatos, V; Boisramé, B
1999-01-01
In the field of controlled drug delivery, most of the reported work is aimed at introducing new systems, or at providing basic information on the critical parameters which affect release profiles in vitro and occasionally in vivo. The situation is totally different when one wants to fulfil the specific requirements imposed by the marketing of a sustained release device to be used in humans or in animals eaten by human beings. The control of the release characteristics is then a difficult challenge. In this work, attempts were made to combine cephradin, a hydrophilic beta-lactam antibiotic, and bioresorbable polymeric matrices of a poly(alpha-hydroxy acid) in the form of microspheres with the aim of delivering the antibiotic to cattle at a dose rate of 4-5 mg/kg/day over a 3-4 days period after i.m. injection. PLAGA aliphatic polyesters were selected because they are already FDA approved as matrices. The solvent evaporation technique using PVA as the emulsion stabilizer was selected because it is efficient and can be extended to an industrial scale. Various experimental conditions were used in order to obtain the highest encapsulation yields compatible with the desired specifications. Decreasing the volume of the aqueous phase and adding a water-miscible organic solvent/non-solvent of cephradin failed. In contrast, microspheres containing up to 30% cephradin were prepared after addition of sodium chloride to the aqueous dispersing phase. The amount of entrapped drug was raised to 40% by decreasing the temperature and the pressure. Preliminary investigations using dogs showed that 20% cephradin microspheres prepared under these conditions extended the presence of cephradin in the blood circulation up to 48 h. Increasing the load led to higher blood concentrations but shorter sustained release. The fact that the microspheres were for cattle limited the volume of the injection and thus the amount of microspheres to be administered. The other limiting factors were related to microsphere morphology.
Wei, Yi; Wang, Yu Xia; Wang, Wei; Ho, Sa V; Qi, Feng; Ma, Guang Hui; Su, Zhi Guo
2012-10-02
The microcosmic mechanisms of protein (recombinant human growth hormone, rhGH) incomplete release and stability from amphiphilic poly(monomethoxypolyethylene glycol-co-D,L-lactide) (mPEG-PLA, PELA) microspheres were investigated. PELA with different hydrophilicities (PELA-1, PELA-2, and PELA-3) based on various ratios of mPEG to PLA were employed to prepare microspheres exhibiting a narrow size distribution using a combined double emulsion and premix membrane emulsification method. The morphology, rhGH encapsulation efficiency, in vitro release profile, and rhGH stability of PELA microspheres during the release were characterized and compared in detail. It was found that increasing amounts of PLA enhanced the encapsulation efficiency of PELA microspheres but reduced both the release rate of rhGH and its stability. Contact angle, atomic force microscope (AFM), and quartz crystal microbalance with dissipation (QCM-D) techniques were first combined to elucidate the microcosmic mechanism of incomplete release by measuring the hydrophilicity of the PELA film and its interaction with rhGH. In addition, the pH change within the microsphere microenvironment was monitored by confocal laser scanning microscopy (CLSM) employing a pH-sensitive dye, which clarified the stability of rhGH during the release. These results suggested that PELA hydrophilicity played an important role in rhGH incomplete release and stability. Thus, the selection of suitable hydrophilic polymers with adequate PEG lengths is critical in the preparation of optimum protein drug sustained release systems. This present work is a first report elucidating the microcosmic mechanisms responsible for rhGH stability and its interaction with the microspheres. Importantly, this research demonstrated the application of promising new experimental methods in investigating the interaction between biomaterials and biomacromolecules, thus opening up a range of exciting potential applications in the biomedical field including drug delivery and tissue regeneration.
Porous PLGA microspheres tailored for dual delivery of biomolecules via layer-by-layer assembly.
Go, Dewi P; Palmer, Jason A; Mitchell, Geraldine M; Gras, Sally L; O'Connor, Andrea J
2015-05-01
Tissue engineering is a complex and dynamic process that requires varied biomolecular cues to promote optimal tissue growth. Consequently, the development of delivery systems capable of sequestering more than one biomolecule with controllable release profiles is a key step in the advancement of this field. This study develops multilayered polyelectrolyte films incorporating alpha-melanocyte stimulating hormone (α-MSH), an anti-inflammatory molecule, and basic fibroblast growth factor (bFGF). The layers were successfully formed on macroporous poly lactic-co-glycolic acid microspheres produced using a combined inkjet and thermally induced phase separation technique. Release profiles could be varied by altering layer properties including the number of layers and concentrations of layering molecules. α-MSH and bFGF were released in a sustained manner and the bioactivity of α-MSH was shown to be preserved using an activated macrophage cell assay in vitro. The system performance was also tested in vivo subcutaneously in rats. The multilayered microspheres reduced the inflammatory response induced by a carrageenan stimulus 6 weeks after implantation compared to the non-layered microspheres without the anti-inflammatory and growth factors, demonstrating the potential of such multilayered constructs for the controlled delivery of bioactive molecules. © 2014 Wiley Periodicals, Inc.
Sustained release of nerve growth factor from biodegradable polymer microspheres.
Camarata, P J; Suryanarayanan, R; Turner, D A; Parker, R G; Ebner, T J
1992-03-01
Although grafted adrenal medullary tissue to the striatum has been used both experimentally and clinically in parkinsonism, there is a definite need to augment long-term survival. Infusion of nerve growth factor (NGF) or implantation of NGF-rich tissue into the area of the graft prolongs survival and induces differentiation into neural-like cells. To provide for prolonged, site-specific delivery of this growth factor to the grafted tissue in a convenient manner, we fabricated biodegradable polymer microspheres of poly(L-lactide)co-glycolide (70:30) containing NGF. Biologically active NGF was released from the microspheres, as assayed by neurite outgrowth in a dorsal root ganglion tissue culture system. Anti-NGF could block this outgrowth. An enzyme-linked immunosorbent assay detected NGF still being released in vitro for longer than 5 weeks. In vivo immunohistochemical studies showed release over a 4.5-week period. This technique should prove useful for incorporating NGF and other growth factors into polymers and delivering proteins and other macromolecules intracerebrally over a prolonged time period. These growth factor-containing polymer microspheres can be used in work aimed at prolonging graft survival, treating experimental Alzheimer's disease, and augmenting peripheral nerve regeneration.
Dorati, Rossella; DeTrizio, Antonella; Genta, Ida; Grisoli, Pietro; Merelli, Alessia; Tomasi, Corrado; Conti, Bice
2016-01-01
The present paper takes into account the DOE application to the preparation process of biodegradable microspheres for osteomyelitis local therapy. With this goal gentamicin loaded polylactide-co-glycolide-copolyethyleneglycol (PLGA-PEG) microspheres were prepared and investigated. Two preparation protocols (o/w and w/o/w) with different process conditions, and three PLGA-PEG block copolymers with different compositions of lactic and glycolic acids and PEG, were tested. A Design Of Experiment (DOE) screening design was applied as an approach to scale up manufacturing step. The results of DOE screening design confirmed that w/o/w technique, the presence of salt and the 15%w/v polymer concentration positively affected the EE% (72.1-97.5%), and span values of particle size distribution (1.03-1.23), while salt addition alone negatively affected the yield process. Process scale up resulted in a decrease of gentamicin EE% that can be attributed to the high volume of water used to remove PVA and NaCl residues. The results of in vitro gentamicin release study show prolonged gentamicin release up to three months from the microspheres prepared with salt addition in the dispersing phase; the behavior being consistent with their highly compact structure highlighted by scanning electron microscopy analysis. The prolonged release of gentamicin is maintained even after embedding the biodegradable microspheres into a thermosetting composite gel made of chitosan and acellular bovine bone matrix (Orthoss® granules), and the microbiologic evaluation demonstrated the efficacy of the gentamicin loaded microspheres on Escherichia coli. The collected results confirm the feasibility of the scale up of microsphere manufacturing process and the high potential of the microparticulate drug delivery system to be used for the local antibiotic delivery to bone.
Bansal, Sanjay; Beg, Sarwar; Asthana, Abhay; Garg, Babita; Asthana, Gyati Shilakari; Kapil, Rishi; Singh, Bhupinder
2016-01-01
The objectives of present studies were to develop the systematically optimized multiple-unit gastroretentive microballoons, i.e. hollow microspheres of itopride hydrochloride (ITH) employing quality by design (QbD)-based approach. Initially, the patient-centric QTPP and CQAs were earmarked, and preliminary studies were conducted to screen the suitable polymer, solvent, solvent ratio, pH and temperature conditions. Microspheres were prepared by non-aqueous solvent evaporation method employing Eudragit S-100. Risk assessment studies carried out by constructing Ishikawa cause-effect fish-bone diagram, and techniques like risk estimation matrix (REM) and failure mode effect analysis (FMEA) facilitated the selection of plausible factors affecting the drug product CQAs, i.e. percent yield, entrapment efficiency (EE) and percent buoyancy. A 3(3) Box-Behnken design (BBD) was employed for optimizing CMAs and CPPs selected during factor screening studies employing Taguchi design, i.e. drug-polymer ratio (X1), stirring temperature (X2) and stirring speed (X3). The hollow microspheres, as per BBD, were evaluated for EE, particle size and drug release characteristics. The optimum formulation was embarked upon using numerical desirability function yielding excellent floatation characteristics along with adequate drug release control. Drug-excipient compatibility studies employing FT-IR, DSC and powder XRD revealed absence of significant interaction among the formulation excipients. The SEM studies on the optimized formulation showed hollow and spherical nature of the prepared microspheres. In vivo X-ray imaging studies in rabbits confirmed the buoyant nature of the hollow microspheres for 8 h in the upper GI tract. In a nutshell, the current investigations report the successful development of gastroretentive floating microspheres for once-a-day administration of ITH.
Hassan, Ahmed Sheikh; Sapin, Anne; Ubrich, Nathalie; Maincent, Philippe; Bolzan, Claire; Leroy, Pierre
2008-10-01
A simple and sensitive high-performance liquid chromatography (HPLC) assay applied to the measurement of ibuprofen in rat plasma has been developed. Two parameters have been investigated to improve ibuprofen detectability using fluorescence detection: variation of mobile phase pH and the use of beta-cyclodextrin (beta-CD). Increasing the pH value from 2.5 to 6.5 and adding 5 mM beta-CD enhanced the fluorescence signal (lambda(exc) = 224 nm; lambda(em) = 290 nm) by 2.5 and 1.3-fold, respectively, when using standards. In the case of plasma samples, only pH variation significantly lowered detection and quantification limits, down to 10 and 35 ng/mL, respectively. Full selectivity was obtained with a single step for plasma treatment, that is, protein precipitation with acidified acetonitrile. The validated method was applied to a pharmacokinetic study of ibuprofen encapsulated in microspheres and subcutaneously administered to rats.
Optical sensors for application in intelligent food-packaging technology
NASA Astrophysics Data System (ADS)
McEvoy, Aisling K.; Von Bueltzingsloewen, Christoph; McDonagh, Colette M.; MacCraith, Brian D.; Klimant, Ingo; Wolfbeis, Otto S.
2003-03-01
Modified Atmosphere Packaged (MAP) food employs a protective gas mixture, which normally contains selected amounts of carbon dioxide (CO2) and oxygen (O2), in order to extend the shelf life of food. Conventional MAP analysis of package integrity involves destructive sampling of packages followed by carbon dioxide and oxygen detection. For quality control reasons, as well as to enhance food safety, the concept of optical on-pack sensors for monitoring the gas composition of the MAP package at different stages of the distribution process is very attractive. The objective of this work was to develop printable formulations of oxygen and carbon dioxide sensors for use in food packaging. Oxygen sensing is achieved by detecting the degree of quenching of a fluorescent ruthenium complex entrapped in a sol-gel matrix. In particular, a measurement technique based on the quenching of the fluorescence decay time, phase fluorometric detection, is employed. A scheme for detecting CO2 has been developed which is compatible with the oxygen detection scheme. It is fluorescence-based and uses the pH-sensitive 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) indicator dye encapsulated in an organically modified silica (ORMOSIL) glass matrix. Dual Luminophore Referencing (DLR) has been employed as an internal referencing scheme, which provides many of the advantages of lifetime-based fluorometric methods. Oxygen cross-sensitivity was minimised by encapsulating the reference luminophore in dense sol-gel microspheres. The sensor performance compared well with standard methods for both oxygen and carbon dioxide detection. The results of preliminary on-pack print trials are presented and a preliminary design of an integrated dual gas optical read-out device is discussed.
Serise, J M; Le Héron, D; Le Héron, G; Bonnet, A; Busquet, J; Basse-Cathalinat, B; Tingaud, R
1982-01-01
The greater proportion of oxygen in the blood circulating in veins of the lower limbs bought to attention the possibility of pre-capillary arteriovenous shunting. The intra-arteriel injection of Technetium labelled serum albumin microspheres enables one to quantify arteriovenous shunts greater than 25 microns. The authors made a comparative study of arterio-venous shunting by blood gases analysis and isotopic techniques in twenty patients. The results seem to deny the existence of arteriovenous shunts in the genesis of essential varicose veins in the lower limb.
High-throughput kinase assays with protein substrates using fluorescent polymer superquenching.
Rininsland, Frauke; Stankewicz, Casey; Weatherford, Wendy; McBranch, Duncan
2005-05-31
High-throughput screening is used by the pharmaceutical industry for identifying lead compounds that interact with targets of pharmacological interest. Because of the key role that aberrant regulation of protein phosphorylation plays in diseases such as cancer, diabetes and hypertension, kinases have become one of the main drug targets. With the exception of antibody-based assays, methods to screen for specific kinase activity are generally restricted to the use of small synthetic peptides as substrates. However, the use of natural protein substrates has the advantage that potential inhibitors can be detected that affect enzyme activity by binding to a site other than the catalytic site. We have previously reported a non-radioactive and non-antibody-based fluorescence quench assay for detection of phosphorylation or dephosphorylation using synthetic peptide substrates. The aim of this work is to develop an assay for detection of phosphorylation of chemically unmodified proteins based on this polymer superquenching platform. Using a modified QTL Lightspeed assay, phosphorylation of native protein was quantified by the interaction of the phosphorylated proteins with metal-ion coordinating groups co-located with fluorescent polymer deposited onto microspheres. The binding of phospho-protein inhibits a dye-labeled "tracer" peptide from associating to the phosphate-binding sites present on the fluorescent microspheres. The resulting inhibition of quench generates a "turn on" assay, in which the signal correlates with the phosphorylation of the substrate. The assay was tested on three different proteins: Myelin Basic Protein (MBP), Histone H1 and Phosphorylated heat- and acid-stable protein (PHAS-1). Phosphorylation of the proteins was detected by Protein Kinase Calpha (PKCalpha) and by the Interleukin -1 Receptor-associated Kinase 4 (IRAK4). Enzyme inhibition yielded IC50 values that were comparable to those obtained using peptide substrates. Statistical parameters that are used in the high-throughput community to determine assay robustness (Z'-value) demonstrate the suitability of this format for high-throughput screening applications for detection of inhibitors of enzyme activity. The QTL Lightspeed protein detection system provides a simple mix and measure "turn on" assay for the detection of kinase activity using natural protein substrates. The platform is robust and allows for identification of inhibitors of kinase activity.
USDA-ARS?s Scientific Manuscript database
A novel technique named multiple-particle tracking (MPT) was used to investigate the micro-structural heterogeneities of Z-trim, a zero calorie cellulosic fiber biopolymer produced from corn hulls. The principle of MPT technique is to monitor the thermally driven motion of inert micro-spheres, which...
Chemical Sintering Generates Uniform Porous Hyaluronic Acid Hydrogels
Cam, Cynthia; Segura, Tatiana
2014-01-01
Implantation of scaffolds for tissue repair has been met with limited success primarily due to the inability to achieve vascularization within the construct. Many strategies have shifted to incorporate pores into these scaffolds to encourage rapid cellular infiltration and subsequent vascular ingrowth. We utilized an efficient chemical sintering technique to create a uniform network of polymethyl methacrylate (PMMA) microspheres for porous hyaluronic acid hydrogel formation. The porous hydrogels generated from chemical sintering possessed comparable pore uniformity and interconnectivity as the commonly used non- and heat sintering techniques. Moreover, similar cell response to the porous hydrogels generated from each sintering approach was observed in cell viability, spreading, proliferation in vitro, as well as, cellular invasion in vivo. We propose chemical sintering of PMMA microspheres using a dilute acetone solution as an alternative method to generating porous hyaluronic acid hydrogels since it requires equal or ten-fold less processing time as the currently used non-sintering or heat sintering technique, respectively. PMID:24120847
Scalable synthesis of mesoporous titania microspheres via spray-drying method.
Pal, Manas; Wan, Li; Zhu, Yongheng; Liu, Yupu; Liu, Yang; Gao, Wenjun; Li, Yuhui; Zheng, Gengfeng; Elzatahry, Ahmed A; Alghamdi, Abdulaziz; Deng, Yonghui; Zhao, Dongyuan
2016-10-01
Mesoporous TiO2 has several potential applications due to its unique electronic and optical properties, although its structures and morphologies are typically difficult to tune because of its uncontrollable and fast sol-gel reaction. In this study we have coupled the template-directed-sol-gel-chemistry with the low-cost, scalable, and environmentally benign aerosol (spray-drying) one-pot preparation technique for the fabrication of hierarchically mesoporous TiO2 microspheres and Fe3O4@mesoporous TiO2-x microspheres in a large scale. Parameters during the pre-hydrolysis and spray-drying treatment were varied to successfully control the bead diameter, morphology, monodispersity, surface area and pore size for improving their effectiveness for better application. Unlike to the previous aerosol synthetic approaches, where mainly quite a high temperature gradient with the strict control of spray-drying precursor concentration is implied, our strategy is lying on comparatively low drying temperature with an additional post-ultrasonication (further hydrolysis and condensation) route of the pre-calcined TiO2 samples. As-synthesized mesoporous microspheres have a size distribution from 500nm to 5μm, specific surface areas ranging from 150 to 162m(2)g(-1) and mean pore sizes of several nanometers (4-6nm). Further Fe3O4@mesoporous TiO2-x microspheres were observed to show remarkable selective phosphopeptide-enrichment activity which might have significant importance in disease diagnosis and other biomedical applications. Copyright © 2016. Published by Elsevier Inc.
Poole, KM; Nelson, CE; Joshi, RV; Martin, JR; Gupta, MK; Haws, SC; Kavanaugh, TE; Skala, MC; Duvall, CL
2014-01-01
A new microparticle-based delivery system was synthesized from reactive oxygen species (ROS)-responsive poly(propylene sulfide) (PPS) and tested for “on demand” antioxidant therapy. PPS is hydrophobic but undergoes a phase change to become hydrophilic upon oxidation and thus provides a useful platform for ROS-demanded drug release. This platform was tested for delivery of the promising anti-inflammatory and antioxidant therapeutic molecule curcumin, which is currently limited in use in its free form due to poor pharmacokinetic properties. PPS microspheres efficiently encapsulated curcumin through oil-in-water emulsion and provided sustained, on demand release that was modulated in vitro by hydrogen peroxide concentration. The cytocompatible, curcumin-loaded microspheres preferentially targeted and scavenged intracellular ROS in activated macrophages, reduced in vitro cell death in the presence of cytotoxic levels of ROS, and decreased tissue-level ROS in vivo in the diabetic mouse hind limb ischemia model of peripheral arterial disease. Interestingly, due to the ROS scavenging behavior of PPS, the blank microparticles also showed inherent therapeutic properties that were synergistic with the effects of curcumin in these assays. Functionally, local delivery of curcumin-PPS microspheres accelerated recovery from hind limb ischemia in diabetic mice, as demonstrated using non-invasive imaging techniques. This work demonstrates the potential for PPS microspheres as a generalizable vehicle for ROS-demanded drug release and establishes the utility of this platform for improving local curcumin bioavailability for treatment of chronic inflammatory diseases. PMID:25522975
Binary breath figures for straightforward and controllable self-assembly of microspherical caps.
Gong, Jianliang; Xu, Bingang; Tao, Xiaoming; Li, Lei
2016-05-11
The intense interest surrounding asymmetrical microparticles originates from their unique anisotropic properties and promising applications. In this work, direct self-assembly of polymeric microspherical caps without the assistance of any additives has been achieved by using low-surface-tension methanol (MeOH) and high-surface-tension water as binary breath figures (BFs). With the evaporation of polystyrene (PS) solution containing low-boiling-point solvent in the binary vapors, the formed MeOH BFs could quickly diffuse into solution, while water BFs tended to remain at the solution surface. This led to the formation of a gradient nonsolvent layer at the vapor/solution interface, which induced the formation of nuclei and guided further asymmetrical growth of polymer particles. After the spontaneous removal of MeOH, water and residual solvent by evaporation, polymeric microspherical caps were left on the substrate. Through controlling the proportion of water introduced by adjusting the ratios of MeOH and water, polymeric microspherical caps with a range of controllable shapes (divided at different positions of a sphere) were successfully obtained. The formation mechanism was explained based on the difference of vapor pressure, surface tension and miscibility between the employed solvents and nonsolvents. A solvent possessing a high vapor pressure, low surface tension and good miscibility with MeOH contributed to the formation of microspherical caps. This flexible, green and straightforward technique is a nondestructive strategy, and avoids complicated work on design, preparation and removal of hard templates and additives.
NASA Astrophysics Data System (ADS)
Neves, Cristina Sofia dos Santos
The work described in this PhD Thesis focuses on the post-processing of optical fibers and their enhancement as sensing element. Since the majority of sensors presented are based in Fabry-Perot interferometers, an historical overview of this category of optical fiber sensors is firstly presented. This review considers the works published since the early years, in the beginning of the 1980s, until the middle of 2015. The incorporation of microcavities at the tip of a single mode fiber was extensively studied, particularly for the measurement of nitrogen and methane gas pressure. These cavities were fabricated using hollow core silica tubes and a hollow core photonic crystal fiber. Following a different approach, the microcavities were incorporated between two sections of single mode fiber. In this case, the low sensitivity to temperature makes these microcavities highly desirable for the measurement of strain at high temperatures. Competences in post-processing techniques such as the chemical etching and the writing of periodical structures in the fiber core by means of an excimer or a femtosecond laser were also acquired in the course of the PhD programme. One of the works consisted in the design and manufacturing of a double clad optical fiber. The refractive index of the inner cladding was higher than the one of the outer cladding and the core. Thus, light was guided in the inner cladding instead of propagating in the core. This situation was overcome by applying chemical etching, thus removing the inner cladding. The core, surrounded by air, was then able to guide light. Two different applications were found for this fiber, as a temperature sensor and as an optical refractometer. In the last, the optical phase changes with the liquid refractive index. Two different types of fiber Bragg gratings were characterized in strain and temperature. Sensing structures obtained through the phase mask technique at the tip of an optical fiber were subjected to chemical etching. In this case, an excimer laser was used. Extremely thin fiber tips were obtained, with an ultra-high sensitivity to strain. The other technique employed to fabricate the fiber Bragg gratings was the point-by-point femtosecond laser inscription. In this case, the sensing elements are very stable at high temperatures and can be used to measure strain in harsh conditions. The employment of optical fiber lasers as sensing elements was also considered in this Thesis. Two laser cavities were studied, one based on the ring configuration and the other based on a figure-of-eight configuration. From these works, the quality of the laser emission, namely the signal-to-noise ratio, the reduced full-width at half maximum and the stability should be highlighted. These characteristics allowed the measurement of different physical parameters, such as strain, temperature and torsion. Lastly, the possibility to use microspheres as sensing elements was considered. Using the electric arc of a fusion splicer, it is possible to create microspheres at the tip of an optical fiber. Furthermore, with this technique it is chains of microspheres can be obtained, constituting Mach-Zehnder-type interferometers which are sensitive to physical parameters like strain and temperature. The preliminary results obtained by introducing silica microspheres in a support structure are also presented. In this case, the sensors were subjected to temperature variations. All the experimental work was combined with the respective theoretical considerations. Many questions have been raised with the course of this PhD, and there are still some without a definite answer. Thus, new research paths can be followed, having their basis grounded in the configurations here presented.
In vitro evaluation of biodegradable microspheres with surface-bound ligands.
Keegan, Mark E; Royce, Sara M; Fahmy, Tarek; Saltzman, W Mark
2006-02-21
Protein ligands were conjugated to the surface of biodegradable microspheres. These microsphere-ligand conjugates were then used in two in vitro model systems to evaluate the effect of conjugated ligands on microsphere behavior. Microsphere retention in agarose columns was increased by ligands on the microsphere surface specific for receptors on the agarose matrix. In another experiment, conjugating the lectin Ulex europaeus agglutinin 1 to the microsphere surface increased microsphere adhesion to Caco-2 monolayers compared to control microspheres. This increase in microsphere adhesion was negated by co-administration of l-fucose, indicating that the increase in adhesion is due to specific interaction of the ligand with carbohydrate receptors on the cell surface. These results demonstrate that the ligands conjugated to the microspheres maintain their receptor binding activity and are present on the microsphere surface at a density sufficient to target the microspheres to both monolayers and three-dimensional matrices bearing complementary receptors.
McGoron, Anthony J; Capille, Michael; Georgiou, Michael F; Sanchez, Pablo; Solano, Juan; Gonzalez-Brito, Manuel; Kuluz, John W
2008-01-01
Background Assessment of cerebral blood flow (CBF) by SPECT could be important in the management of patients with severe traumatic brain injury (TBI) because changes in regional CBF can affect outcome by promoting edema formation and intracranial pressure elevation (with cerebral hyperemia), or by causing secondary ischemic injury including post-traumatic stroke. The purpose of this study was to establish an improved method for evaluating regional CBF changes after TBI in piglets. Methods The focal effects of moderate traumatic brain injury (TBI) on cerebral blood flow (CBF) by SPECT cerebral blood perfusion (CBP) imaging in an animal model were investigated by parallelized statistical techniques. Regional CBF was measured by radioactive microspheres and by SPECT 2 hours after injury in sham-operated piglets versus those receiving severe TBI by fluid-percussion injury to the left parietal lobe. Qualitative SPECT CBP accuracy was assessed against reference radioactive microsphere regional CBF measurements by map reconstruction, registration and smoothing. Cerebral hypoperfusion in the test group was identified at the voxel level using statistical parametric mapping (SPM). Results A significant area of hypoperfusion (P < 0.01) was found as a response to the TBI. Statistical mapping of the reference microsphere CBF data confirms a focal decrease found with SPECT and SPM. Conclusion The suitability of SPM for application to the experimental model and ability to provide insight into CBF changes in response to traumatic injury was validated by the SPECT SPM result of a decrease in CBP at the left parietal region injury area of the test group. Further study and correlation of this characteristic lesion with long-term outcomes and auxiliary diagnostic modalities is critical to developing more effective critical care treatment guidelines and automated medical imaging processing techniques. PMID:18312639
McGoron, Anthony J; Capille, Michael; Georgiou, Michael F; Sanchez, Pablo; Solano, Juan; Gonzalez-Brito, Manuel; Kuluz, John W
2008-02-29
Assessment of cerebral blood flow (CBF) by SPECT could be important in the management of patients with severe traumatic brain injury (TBI) because changes in regional CBF can affect outcome by promoting edema formation and intracranial pressure elevation (with cerebral hyperemia), or by causing secondary ischemic injury including post-traumatic stroke. The purpose of this study was to establish an improved method for evaluating regional CBF changes after TBI in piglets. The focal effects of moderate traumatic brain injury (TBI) on cerebral blood flow (CBF) by SPECT cerebral blood perfusion (CBP) imaging in an animal model were investigated by parallelized statistical techniques. Regional CBF was measured by radioactive microspheres and by SPECT 2 hours after injury in sham-operated piglets versus those receiving severe TBI by fluid-percussion injury to the left parietal lobe. Qualitative SPECT CBP accuracy was assessed against reference radioactive microsphere regional CBF measurements by map reconstruction, registration and smoothing. Cerebral hypoperfusion in the test group was identified at the voxel level using statistical parametric mapping (SPM). A significant area of hypoperfusion (P < 0.01) was found as a response to the TBI. Statistical mapping of the reference microsphere CBF data confirms a focal decrease found with SPECT and SPM. The suitability of SPM for application to the experimental model and ability to provide insight into CBF changes in response to traumatic injury was validated by the SPECT SPM result of a decrease in CBP at the left parietal region injury area of the test group. Further study and correlation of this characteristic lesion with long-term outcomes and auxiliary diagnostic modalities is critical to developing more effective critical care treatment guidelines and automated medical imaging processing techniques.
Morphological resonances for multicomponent immunoassays
NASA Astrophysics Data System (ADS)
Whitten, W. B.; Shapiro, M. J.; Ramsey, J. M.; Bronk, B. V.
1995-06-01
An immunoassay technique capable of detecting and identifying a number of species of microorganisms in a single analysis is described. The method uses optical-resonance size discrimination of microspheres to identify antibodies to which stained microorganisms are bound.
Church, Cody; Mawko, George; Archambault, John Paul; Lewandowski, Robert; Liu, David; Kehoe, Sharon; Boyd, Daniel; Abraham, Robert; Syme, Alasdair
2018-02-01
Radiopaque microspheres may provide intraprocedural and postprocedural feedback during transarterial radioembolization (TARE). Furthermore, the potential to use higher resolution x-ray imaging techniques as opposed to nuclear medicine imaging suggests that significant improvements in the accuracy and precision of radiation dosimetry calculations could be realized for this type of therapy. This study investigates the absorbed dose kernel for novel radiopaque microspheres including contributions of both short and long-lived contaminant radionuclides while concurrently quantifying the self-shielding of the glass network. Monte Carlo simulations using EGSnrc were performed to determine the dose kernels for all monoenergetic electron emissions and all beta spectra for radionuclides reported in a neutron activation study of the microspheres. Simulations were benchmarked against an accepted 90 Y dose point kernel. Self-shielding was quantified for the microspheres by simulating an isotropically emitting, uniformly distributed source, in glass and in water. The ratio of the absorbed doses was scored as a function of distance from a microsphere. The absorbed dose kernel for the microspheres was calculated for (a) two bead formulations following (b) two different durations of neutron activation, at (c) various time points following activation. Self-shielding varies with time postremoval from the reactor. At early time points, it is less pronounced due to the higher energies of the emissions. It is on the order of 0.4-2.8% at a radial distance of 5.43 mm with increased size from 10 to 50 μm in diameter during the time that the microspheres would be administered to a patient. At long time points, self-shielding is more pronounced and can reach values in excess of 20% near the end of the range of the emissions. Absorbed dose kernels for 90 Y, 90m Y, 85m Sr, 85 Sr, 87m Sr, 89 Sr, 70 Ga, 72 Ga, and 31 Si are presented and used to determine an overall kernel for the microspheres based on weighted activities. The shapes of the absorbed dose kernels are dominated at short times postactivation by the contributions of 70 Ga and 72 Ga. Following decay of the short-lived contaminants, the absorbed dose kernel is effectively that of 90 Y. After approximately 1000 h postactivation, the contributions of 85 Sr and 89 Sr become increasingly dominant, though the absorbed dose-rate around the beads drops by roughly four orders of magnitude. The introduction of high atomic number elements for the purpose of increasing radiopacity necessarily leads to the production of radionuclides other than 90 Y in the microspheres. Most of the radionuclides in this study are short-lived and are likely not of any significant concern for this therapeutic agent. The presence of small quantities of longer lived radionuclides will change the shape of the absorbed dose kernel around a microsphere at long time points postadministration when activity levels are significantly reduced. © 2017 American Association of Physicists in Medicine.
Composition and structure of calcium aluminosilicate microspheres
NASA Astrophysics Data System (ADS)
Sharonova, O. M.; Oreshkina, N. A.; Zhizhaev, A. M.
2017-06-01
The composition was studied of calcium aluminosilicate microspheres of three morphological types in high-calcium fly ash from combustion of brown coal from the Kansk-Achinsk basin in slag-tap boilers at temperatures from 1400 to 1500°C and sampled in the first field of electrostatic precipitators at the Krasnoyarsk Cogeneration Power Station no. 2 (TETs-2). Gross compositions and the composition of local areas were determined using a scanning electron microscopy technique and an energy-dispersive analysis with full mapping of globules. With a high content of basic oxides O ox (68 to 79 wt %) and a low content of acid oxides K ox (21 to 31 wt %), type 1 microspheres are formed. They consist of heterogeneous areas having a porous structure and crystalline components in which the content of CaO, SiO2, or Al2O3 differs by two to three times and the content of MgO differs by seven times. With a lower content of O ox (55 to 63 wt %) and an elevated content of K ox (37 to 45 wt %), type 2 microspheres are formed. They are more homogeneous in the composition and structure and consist of similar crystalline components. Having a close content of O ox (46 to 53 wt %) and K ox (47 to 54 wt %), type 3 microspheres, which are a dense matter consisting of amorphous substance with submicron- and nanostructure of crystalline components, are formed. The basic precursor in formation of high-calcium aluminosilicate microspheres is calcium from the organomineral matter of coals with various contribution of Mg, Fe, S, or Na from the coal organic matter and Al, Fe, S, or Si in the form of single mineral inclusions in a coal particle. On the basis of the available data, the effect was analyzed of the composition of a CaO-MgO-Al2O3-SiO2-FeO system on the melting and viscous properties of the matter in microspheres and formation of globules of different morphology. The results of this analysis will help to find a correlation with properties of microspheres in their use as functional microaggregates in cement or polymeric composite materials, or in the production of ceramic membranes or zeolite sorbents.
Multimodal flexible cystoscopy for creating co-registered panoramas of the bladder urothelium
NASA Astrophysics Data System (ADS)
Seibel, Eric J.; Soper, Timothy D.; Burkhardt, Matthew R.; Porter, Michael P.; Yoon, W. Jong
2012-02-01
Bladder cancer is the most expensive cancer to treat due to the high rate of recurrence. Though white light cystoscopy is the gold standard for bladder cancer surveillance, the advent of fluorescence biomarkers provides an opportunity to improve sensitivity for early detection and reduced recurrence resulting from more accurate excision. Ideally, fluorescence information could be combined with standard reflectance images to provide multimodal views of the bladder wall. The scanning fiber endoscope (SFE) of 1.2mm in diameter is able to acquire wide-field multimodal video from a bladder phantom with fluorescence cancer "hot-spots". The SFE generates images by scanning red, green, and blue (RGB) laser light and detects the backscatter signal for reflectance video of 500-line resolution at 30 frames per second. We imaged a bladder phantom with painted vessels and mimicked fluorescent lesions by applying green fluorescent microspheres to the surface. By eliminating the green laser illumination, simultaneous reflectance and fluorescence images can be acquired at the same field of view, resolution, and frame rate. Moreover, the multimodal SFE is combined with a robotic steering mechanism and image stitching software as part of a fully automated bladder surveillance system. Using this system, the SFE can be reliably articulated over the entire 360° bladder surface. Acquired images can then be stitched into a multimodal 3D panorama of the bladder using software developed in our laboratory. In each panorama, the fluorescence images are exactly co-registered with RGB reflectance.
Chen, Hua-Yan; Wei, Jing-Ru; Pan, Jiong-Xiu; Zhang, Wei; Dang, Fu-Quan; Zhang, Zhi-Qi; Zhang, Jing
2017-05-15
5-hydroxymethylcytosine (5hmC) is the sixth base of DNA. It is involved in active DNA demethylation and can be a marker of diseases such as cancer. In this study, we developed a simple and sensitive 2-(4-boronophenyl)quinoline-4-carboxylic acid modified poly (glycidyl methacrylate (PBAQA-PGMA) fluorescent probe to detect the 5hmC content of genomic DNA based on T4 β-glucosyltransferase-catalyzed glucosylation of 5hmC. The fluorescence-enhanced intensity recorded from the DNA sample was proportional to its 5-hydroxymethylcytosine content and could be quantified by fluorescence spectrophotometry. The developed probe showed good detection sensitivity and selectivity and a good linear relationship between the fluorescence intensity and the concentration of 5 hmC within a 0-100nM range. Compared with other fluorescence detection methods, this method not only could determine trace amounts of 5 hmC from genomic DNA but also could eliminate the interference of fluorescent dyes and the need for purification. It also could avoid multiple labeling. Because the PBAQA-PGMA probe could enrich the content of glycosyl-5-hydroxymethyl-2-deoxycytidine from a complex ground substance, it will broaden the linear detection range and improve sensitivity. The limit of detection was calculated to be 0.167nM after enrichment. Furthermore, the method was successfully used to detect 5-hydroxymethylcytosine from mouse tissues. Copyright © 2016 Elsevier B.V. All rights reserved.
Encapsulation of Naproxen in Lipid-Based Matrix Microspheres: Characterization and Release Kinetics
Bhoyar, PK; Morani, DO; Biyani, DM; Umekar, MJ; Mahure, JG; Amgaonkar, YM
2011-01-01
The objective of this study was to microencapsulate the anti-inflammatory drug (naproxen) to provide controlled release and minimizing or eliminating local side effect by avoiding the drug release in the upper gastrointestinal track. Naproxen was microencapsulated with lipid-like carnauba wax, hydrogenated castor oil using modified melt dispersion (modified congealable disperse phase encapsulation) technique. Effect of various formulation and process variables such as drug-lipid ratio, concentration of modifier, concentration of dispersant, stirring speed, stirring time, temperature of external phase, on evaluatory parameters such as size, entrapment efficiency, and in vitro release of naproxen were studied. The microspheres were characterized for particle size, scanning electron microscopy (SEM), FT-IR spectroscopy, drug entrapment efficiency, in vitro release studies, for in vitro release kinetics. The shape of microspheres was found to be spherical by SEM. The drug entrapment efficiency of various batches of microspheres was found to be ranging from 60 to 90 %w/w. In vitro drug release studies were carried out up to 24 h in pH 7.4 phosphate buffer showing 50-65% drug release. In vitro drug release from all the batches showed better fitting with the Korsmeyer-Peppas model, indicating the possible mechanism of drug release to be by diffusion and erosion of the lipid matrix. PMID:21731354
Encapsulation of naproxen in lipid-based matrix microspheres: characterization and release kinetics.
Bhoyar, P K; Morani, D O; Biyani, D M; Umekar, M J; Mahure, J G; Amgaonkar, Y M
2011-04-01
The objective of this study was to microencapsulate the anti-inflammatory drug (naproxen) to provide controlled release and minimizing or eliminating local side effect by avoiding the drug release in the upper gastrointestinal track. Naproxen was microencapsulated with lipid-like carnauba wax, hydrogenated castor oil using modified melt dispersion (modified congealable disperse phase encapsulation) technique. Effect of various formulation and process variables such as drug-lipid ratio, concentration of modifier, concentration of dispersant, stirring speed, stirring time, temperature of external phase, on evaluatory parameters such as size, entrapment efficiency, and in vitro release of naproxen were studied. The microspheres were characterized for particle size, scanning electron microscopy (SEM), FT-IR spectroscopy, drug entrapment efficiency, in vitro release studies, for in vitro release kinetics. The shape of microspheres was found to be spherical by SEM. The drug entrapment efficiency of various batches of microspheres was found to be ranging from 60 to 90 %w/w. In vitro drug release studies were carried out up to 24 h in pH 7.4 phosphate buffer showing 50-65% drug release. In vitro drug release from all the batches showed better fitting with the Korsmeyer-Peppas model, indicating the possible mechanism of drug release to be by diffusion and erosion of the lipid matrix.
Hutchens, Thomas C; Darafsheh, Arash; Fardad, Amir; Antoszyk, Andrew N; Ying, Howard S; Astratov, Vasily N; Fried, Nathaniel M
2012-06-01
Ophthalmic surgery may benefit from use of more precise fiber delivery systems during laser surgery. Some current ophthalmic surgical techniques rely on tedious mechanical dissection of tissue layers. In this study, chains of sapphire microspheres integrated into a hollow waveguide distal tip are used for erbium:YAG laser ablation studies in contact mode with ophthalmic tissues, ex vivo. The laser's short optical penetration depth combined with the small spot diameters achieved with this fiber probe may provide more precise tissue removal. One-, three-, and five-microsphere chain structures were characterized, resulting in FWHM diameters of 67, 32, and 30 μm in air, respectively, with beam profiles comparable to simulations. Single Er:YAG pulses of 0.1 mJ and 75-μs duration produced ablation craters with average diameters of 44, 30, and 17 μm and depths of 26, 10, and 8 μm, for one-, three-, and five-sphere structures, respectively. Microsphere chains produced spatial filtering of the multimode Er:YAG laser beam and fiber, providing spot diameters not otherwise available with conventional fiber systems. Because of the extremely shallow treatment depth, compact focused beam, and contact mode operation, this probe may have potential for use in dissecting epiretinal membranes and other ophthalmic tissues without damaging adjacent retinal tissue.
Method of fabrication of electrodes and electrolytes
Jankowski, Alan F.; Morse, Jeffrey D.
2004-01-06
Fuel cell stacks contain an electrolyte layer surrounded on top and bottom by an electrode layer. Porous electrodes are prepared which enable fuel and oxidant to easily flow to the respective electrode-electrolyte interface without the need for high temperatures or pressures to assist the flow. Rigid, inert microspheres in combination with thin-film metal deposition techniques are used to fabricate porous anodes, cathodes, and electrolytes. Microshperes contained in a liquid are randomly dispersed onto a host structure and dried such that the microsperes remain in position. A thin-film deposition technique is subsequently employed to deposit a metal layer onto the microsperes. After such metal layer deposition, the microspheres are removed leaving voids, i.e. pores, in the metal layer, thus forming a porous electrode. Successive repetitions of the fabrication process result in the formation of a continuous fuel cell stack. Such stacks may produce power outputs ranging from about 0.1 Watt to about 50 Watts.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Randall, N.J.; Beard, R.W.; Sutherland, I.A.
1988-03-01
Data obtained from a thermal system capable of measuring changes in organ temperature as well as tissue thermal clearance in the uterus and vagina have been compared with blood flow measured continuously with a transit-time ultrasound volume-flow sensor placed around the common internal iliac artery and intermittently with radioactive microspheres in the chronically instrumented nonpregnant sheep. Temperature changes in both the uterus and the vagina correlated well with blood flow changes measured by both techniques after intravenous administration of estradiol or norepinephrine. Thermal clearance did not correlate well with blood flow in the vagina or uterus. These methods may havemore » value in the investigation of blood flow patterns in various clinical situations such as the pelvic pain syndrome and early pregnancy.« less
Enhanced optical rotation and diminished depolarization in diffusive scattering from a chiral liquid
NASA Astrophysics Data System (ADS)
Silverman, M. P.; Strange, Wayne; Badoz, J.; Vitkin, I. A.
1996-02-01
Optical rotation and degree of polarization of linearly polarized light were observed by forward, lateral, and back scattering from solutions of D-glucose containing a dispersion of micron-size polystyrene spheres. Rotations increased linearly with glucose concentration at a rate determined by the microsphere concentration and were large even at optical thicknesses sufficiently great to extinguish transmission of the incident beam. Depolarization of light with increasing microsphere concentration occurred at a much slower rate in chiral glucose solution than in pure water. These experiments suggest new possibilities for studying turbid chiral media for which light transmission and specular reflection techniques are inappropriate.
Walt, David R.; Mandal, Tarun K.; Fleming, Michael S.
2004-04-13
The invention features core-shell microsphere compositions, hollow polymeric microspheres, and methods for making the microspheres. The microspheres are characterized as having a polymeric shell with consistent shell thickness.
Simultaneous Determination of Glass Transition Temperatures of Several Polymers
He, Jiang; Liu, Wei; Huang, Yao-Xiong
2016-01-01
Aims A simple and easy optical method is proposed for the determination of glass transition temperature (Tg) of polymers. Methods & Results Tg was determined using the technique of microsphere imaging to monitor the variation of the refractive index of polymer microsphere as a function of temperature. It was demonstrated that the method can eliminate most thermal lag and has sensitivity about six fold higher than the conventional method in Tg determination. So the determined Tg is more accurate and varies less with cooling/heating rate than that obtained by conventional methods. The most attractive character of the method is that it can simultaneously determine the Tg of several polymers in a single experiment, so it can greatly save experimental time and heating energy. Conclusion The method is not only applicable for polymer microspheres, but also for the materials with arbitrary shapes. Therefore, it is expected to be broadly applied to different fundamental researches and practical applications of polymers. PMID:26985670
Manufacture of poly(methyl methacrylate) microspheres using membrane emulsification
Bux, Jaiyana; Manga, Mohamed S.; Hunter, Timothy N.
2016-01-01
Accurate control of particle size at relatively narrow polydispersity remains a key challenge in the production of synthetic polymer particles at scale. A cross-flow membrane emulsification (XME) technique was used here in the preparation of poly(methyl methacrylate) microspheres at a 1–10 l h−1 scale, to demonstrate its application for such a manufacturing challenge. XME technology has previously been shown to provide good control over emulsion droplet sizes with careful choice of the operating conditions. We demonstrate here that, for an appropriate formulation, equivalent control can be gained for a precursor emulsion in a batch suspension polymerization process. We report here the influence of key parameters on the emulsification process; we also demonstrate the close correlation in size between the precursor emulsion and the final polymer particles. Two types of polymer particle were produced in this work: a solid microsphere and an oil-filled matrix microcapsule. This article is part of the themed issue ‘Soft interfacial materials: from fundamentals to formulation’. PMID:27298430
Ternary particles for effective vaccine delivery to the pulmonary system
NASA Astrophysics Data System (ADS)
Terry, Treniece La'shay
Progress in the fields of molecular biology and genomics has provided great insight into the pathogenesis of disease and the defense mechanisms of the immune system. This knowledge has lead to the classification of an array of abnormal genes, for which, treatment relies on cellular expression of proteins. The utility of DNA-based vaccines hold great promise for the treatment of genetically based and infectious diseases, which ranges from hemophilia, cystic fibrosis, and HIV. Synthetic delivery systems consisting of cationic polymers, such as polyethylenimine (PEI), are capable of condensing DNA into compact structures, maximizing cellular uptake of DNA and yielding high levels of protein expression. To date, short term expression is a major obstacle in the development of gene therapies and has halted their expansion in clinical applications. This study intends to develop a sustained release vaccine delivery system using PLA-PEG block copolymers encapsulating PEI:DNA polyplexes. To enhance the effectiveness of such DNA-based vaccines, resident antigen presenting cells, macrophages and dendritic cells, will be targeted within the alveoli regions of the lungs. Porous microspheres will be engineered with aerodynamic properties capable of achieving deep lung deposition. A fabrication technique using concentric nozzles will be developed to produce porous microspheres. It was observed that modifications in the dispersed to continuous phase ratios have the largest influence on particle size distributions, release rates and encapsulation efficiency which ranged form 80--95% with fourteen days of release. Amphiphilic block copolymers were also used to fabricate porous microspheres. The confirmation of PEG within the biodegradable polymer backbone was found to have a tremendous impact on the microsphere morphology and encapsulation efficiency which varied from 50--90%. Porous microspheres were capable of providing sustained gene expression when tested in vitro using the luciferase reporter gene plasmid DNA. Prolonged expression was obtained for 9 days. PLGA and PLA-PEG microspheres were administered in vivo by intra-tracheal instillation and produced an acute inflammatory response, as observed from the large presence of neutrophils. The response using PLA-PEG microspheres yielded a lower total cell count signifying the incorporation of PEG into the copolymer backbone enhances the biocompatibility of the delivery system.
Zhang, Huan; Chen, Lin; Li, Longfei; Yang, Yongzhen; Liu, Xuguang
2016-12-01
To investigate the influence of magnetic-thermosensitive carbon microspheres (MTCMSs) as a targeting drug carrier on serum albumins in vitro, in this study, bovine serum albumin (BSA) was chosen as a template protein to explore the interaction between serum proteins and MTCMSs. Fluorescence spectrophotometry, ultraviolet-visible absorbance (UV-vis) spectrophotometry and circular dichroism spectrometry were used to investigate the interaction between MTCMSs and BSA. Results indicate that BSA interacts with MTCMSs and the fluorescence intensity of BSA is quenched by 50% in a static quenching at 310 K when the concentration of MTCMSs reaches 30 mg/L. Thermodynamic parameters including free energy change (△G θ ), enthalpy change (△H θ ) and entropy change (△S θ ) were calculated. The results (△G θ < 0, △H θ < 0 and △S θ > 0) suggest a spontaneous process and the formation of a hydrogen bond between MTCMSs and BSA. UV-vis measurements reveal that the micro-environment of an amino acid residue is altered in the presence of MTCMSs. The α-helix content of BSA decreases by 4% and the β-sheet content increases by 3.2% with increasing concentrations of MTCMSs to 30 mg/L, illustrating a change in the skeletal structure of BSA. These results demonstrate that MTCMSs as a targeting drug carrier impact the structure of serum albumins. This work provides not only a theoretical basis of BSA adsorption onto MTCMSs, but also an understanding of safe drug carriers in biomedicine. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.
Adsorption of poly(ethylene glycol)-modified ribonuclease A to a poly(lactide-co-glycolide) surface.
Daly, Susan M; Przybycien, Todd M; Tilton, Robert D
2005-06-30
Protein adsorption is a source of variability in the release profiles of therapeutic proteins from biodegradable microspheres. We employ optical reflectometry and total internal reflection fluorescence to explore the extent and kinetics of ribonuclease A (RNase A) adsorption to spin-cast films of poly(lactide-co-glycolide) (PLG) and, in particular, to determine how covalent grafting of polyethylene glycol (PEG) to RNase A affects adsorption. Adsorption kinetics on PLG surfaces are surface-limited for RNase A but transport-limited for unconjugated PEG homopolymers and for PEG-modified RNase A, indicating that PEG anchors the conjugates to the surface during the transport-limited regime. PEG modification of RNase A decreases the total number of adsorbed molecules per unit area but increases the areal surface coverage because the grafted PEG chains exclude additional surface area. Total internal reflection fluorescence-based exchange measurements show that there is no exchange between adsorbed and solution-phase protein molecules. This indicates an unusually tenacious adsorption. Streaming current measurements indicate that the zeta potential of the PLG surface becomes increasingly negative as the film is exposed to water for several weeks, as expected. Aging of the PLG surface results in increased adsorption of unmodified RNase A but decreased adsorption of unconjugated PEG homopolymers and of PEG-RNase A conjugates, relative to the extent of adsorption on freshly prepared PLG surfaces. Adsorption results correlate well with an increase in the rate, total extent and preservation of bioactivity of RNase A released from PLG microspheres for the PEG-modified version of RNase A.
Biagini, R E; Sammons, D L; Smith, J P; Page, E H; Snawder, J E; Striley, C A F; MacKenzie, B A
2004-08-01
To evaluate potential exposure to Bacillis anthracis (Ba) spores in sampling/decontamination workers in the aftermath of an anthrax terror attack. Fifty six serum samples were obtained from workers involved in environmental sampling for Ba spores at the American Media, Inc. (AMI) building in Boca Raton, FL after the anthrax attack there in October 2001. Nineteen sera were drawn from individuals both pre-entry and several weeks after entrance into the building. Nine sera each were drawn from unique individuals at the pre-entry and follow up blood draws. Thirteen donor control sera were also evaluated. Individuals were surveyed for Ba exposure by measurement of serum Ba anti-protective antigen (PA) specific IgG antibodies using a newly developed fluorescent covalent microsphere immunoassay (FCMIA). Four sera gave positive anti-PA IgG results (defined as anti-PA IgG concentrations > or = the mean microg/ml anti-PA IgG from donor control sera (n = 13 plus 2 SD which were also inhibited > or = 85% when the serum was pre-adsorbed with PA). The positive sera were the pre-entry and follow up samples of two workers who had received their last dose of anthrax vaccine in 2000. It appears that the sampling/decontamination workers of the present study either had insufficient exposure to Ba spores to cause the production of anti-PA IgG antibodies or they were exposed to anthrax spores without producing antibody. The FCMIA appears to be a fast, sensitive, accurate, and precise method for the measurement of anti-PA IgG antibodies.
Wöhl-Bruhn, S; Heim, E; Schwoerer, A; Bertz, A; Harling, S; Menzel, H; Schilling, M; Ludwig, F; Bunjes, H
2012-10-15
Hydrogels are under investigation as long term delivery systems for biomacromolecules as active pharmaceutical ingredients. The release behavior of hydrogels can be tailored during the fabrication process. This study investigates the applicability of fluxgate magnetorelaxometry (MRX) as a tool to characterize the release properties of such long term drug delivery depots. MRX is based on the use of superparamagnetic core-shell nanoparticles as model substances. The feasibility of using superparamagnetic nanoparticles to study the degradation of and the associated release from hydrogel cylinders and hydrogel microspheres was a major point of interest. Gels prepared from two types of photo crosslinkable polymers based on modified hydroxyethylstarch, specifically hydroxyethyl starch-hydroxyethyl methacrylate (HES-HEMA) and hydroxyethyl starch-polyethylene glycol methacrylate (HES-P(EG)(6)MA), were analyzed. MRX analysis of the incorporated nanoparticles allowed to evaluate the influence of different crosslinking conditions during hydrogel production as well as to follow the increase in nanoparticle mobility as a result of hydrogel degradation during release studies. Conventional release studies with fluorescent markers (half-change method) were performed for comparison. MRX with superparamagnetic nanoparticles as model substances is a promising method to analyze pharmaceutically relevant processes such as the degradation of hydrogel drug carrier systems. In contrast to conventional release experiments MRX allows measurements in closed vials (reducing loss of sample and sampling errors), in opaque media and at low magnetic nanoparticle concentrations. Magnetic markers possess a better long-term stability than fluorescent ones and are thus also promising for the use in in vivo studies. Copyright © 2012 Elsevier B.V. All rights reserved.
Mathematical model of a DIC position sensing system within an optical trap
NASA Astrophysics Data System (ADS)
Wulff, Kurt D.; Cole, Daniel G.; Clark, Robert L.
2005-08-01
The quantitative study of displacements and forces of motor proteins and processes that occur at the microscopic level and below require a high level of sensitivity. For optical traps, two techniques for position sensing have been accepted and used quite extensively: quadrant photodiodes and an interferometric position sensing technique based on DIC imaging. While quadrant photodiodes have been studied in depth and mathematically characterized, a mathematical characterization of the interferometric position sensor has not been presented to the authors' knowledge. The interferometric position sensing method works off of the DIC imaging capabilities of a microscope. Circularly polarized light is sent into the microscope and the Wollaston prism used for DIC imaging splits the beam into its orthogonal components, displacing them by a set distance determined by the user. The distance between the axes of the beams is set so the beams overlap at the specimen plane and effectively share the trapped microsphere. A second prism then recombines the light beams and the exiting laser light's polarization is measured and related to position. In this paper we outline the mathematical characterization of a microsphere suspended in an optical trap using a DIC position sensing method. The sensitivity of this mathematical model is then compared to the QPD model. The mathematical model of a microsphere in an optical trap can serve as a calibration curve for an experimental setup.
A reproducible accelerated in vitro release testing method for PLGA microspheres.
Shen, Jie; Lee, Kyulim; Choi, Stephanie; Qu, Wen; Wang, Yan; Burgess, Diane J
2016-02-10
The objective of the present study was to develop a discriminatory and reproducible accelerated in vitro release method for long-acting PLGA microspheres with inner structure/porosity differences. Risperidone was chosen as a model drug. Qualitatively and quantitatively equivalent PLGA microspheres with different inner structure/porosity were obtained using different manufacturing processes. Physicochemical properties as well as degradation profiles of the prepared microspheres were investigated. Furthermore, in vitro release testing of the prepared risperidone microspheres was performed using the most common in vitro release methods (i.e., sample-and-separate and flow through) for this type of product. The obtained compositionally equivalent risperidone microspheres had similar drug loading but different inner structure/porosity. When microsphere particle size appeared similar, porous risperidone microspheres showed faster microsphere degradation and drug release compared with less porous microspheres. Both in vitro release methods investigated were able to differentiate risperidone microsphere formulations with differences in porosity under real-time (37 °C) and accelerated (45 °C) testing conditions. Notably, only the accelerated USP apparatus 4 method showed good reproducibility for highly porous risperidone microspheres. These results indicated that the accelerated USP apparatus 4 method is an appropriate fast quality control tool for long-acting PLGA microspheres (even with porous structures). Copyright © 2015 Elsevier B.V. All rights reserved.
Characterization of a Polyamine Microsphere and Its Adsorption for Protein
Wang, Feng; Liu, Pei; Nie, Tingting; Wei, Huixian; Cui, Zhenggang
2013-01-01
A novel polyamine microsphere, prepared from the water-in-oil emulsion of polyethylenimine, was characterized. The investigation of scanning electron microscopy showed that the polyamine microsphere is a regular ball with a smooth surface. The diameter distribution of the microsphere is 0.37–4.29 μm. The isoelectric point of the microsphere is 10.6. The microsphere can adsorb proteins through the co-effect of electrostatic and hydrophobic interactions. Among the proteins tested, the highest value of adsorption of microsphere, 127.8 mg·g−1 microsphere, was obtained with lipase. In comparison with other proteins, the hydrophobic force is more important in promoting the adsorption of lipase. The microsphere can preferentially adsorb lipase from an even mixture of proteins. The optimum temperature and pH for the selective adsorption of lipase by the microsphere was 35 °C and pH 7.0. PMID:23344018
Liu, Dong; Deng, Jianping; Yang, Wantai
2014-01-01
The first synthesis of porous, optically active, magnetic Fe3 O4 @poly(N-acryloyl-leucine) inverse core/shell composite microspheres is reported, in which the core is constructed of chiral polymer and the shell is constructed of Fe3 O4 NPs. The microspheres integrate three significant concepts, "porosity", "chirality", and "magneticity", in one single microspheric entity. The microspheres consist of Fe3 O4 nanoparticles and porous optically active microspheres, and thus combine the advantages of both magnetic nanoparticles and porous optically active microspheres. The pore size and specific surface area of the microspheres are characterized by N2 adsorption, from which it is found that the composite microspheres possess a desirable porous structure. Circular dichroism and UV-vis absorption spectroscopy measurements demonstrate that the microspheres exhibit the expected optical activity. The microspheres also have high saturation magnetization of 14.7 emu g(-1) and rapid magnetic responsivity. After further optimization, these novel microspheres may potentially find applications in areas such as asymmetric catalysis, chiral adsorption, etc. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
NASA Astrophysics Data System (ADS)
Lin, Bo; Li, Qiufeng; Liu, Baodong; Zhang, Sen; Deng, Chao
2016-04-01
Biochemistry-directed synthesis of functional nanomaterials has attracted great interest in energy storage, catalysis and other applications. The unique ability of biological systems to guide molecule self-assembling facilitates the construction of distinctive architectures with desirable physicochemical characteristics. Herein, we report a biochemistry-directed ``bottom-up'' approach to construct hollow porous microspheres of polyanion materials for sodium ion batteries. Two kinds of polyanions, i.e. Na3V2(PO4)3 and Na3.12Fe2.44(P2O7)2, are employed as cases in this study. The microalgae cell realizes the formation of a spherical ``bottom'' bio-precursor. Its tiny core is subjected to destruction and its tough shell tends to carbonize upon calcination, resulting in the hollow porous microspheres for the ``top'' product. The nanoscale crystals of the polyanion materials are tightly enwrapped by the highly-conductive framework in the hollow microsphere, resulting in the hierarchical nano-microstructure. The whole formation process is disclosed as a ``bottom-up'' mechanism. Moreover, the biochemistry-directed self-assembly process is confirmed to play a crucial role in the construction of the final architecture. Taking advantage of the well-defined hollow-microsphere architecture, the abundant interior voids and the highly-conductive framework, polyanion materials show favourable sodium-intercalation kinetics. Both materials are capable of high-rate long-term cycling. After five hundred cycles at 20 C and 10 C, Na3V2(PO4)3 and Na3.12Fe2.44(P2O7)2 retain 96.2% and 93.1% of the initial capacity, respectively. Therefore, the biochemistry-directed technique provides a low-cost, highly-efficient and widely applicable strategy to produce high-performance polyanion-based cathodes for sodium ion batteries.Biochemistry-directed synthesis of functional nanomaterials has attracted great interest in energy storage, catalysis and other applications. The unique ability of biological systems to guide molecule self-assembling facilitates the construction of distinctive architectures with desirable physicochemical characteristics. Herein, we report a biochemistry-directed ``bottom-up'' approach to construct hollow porous microspheres of polyanion materials for sodium ion batteries. Two kinds of polyanions, i.e. Na3V2(PO4)3 and Na3.12Fe2.44(P2O7)2, are employed as cases in this study. The microalgae cell realizes the formation of a spherical ``bottom'' bio-precursor. Its tiny core is subjected to destruction and its tough shell tends to carbonize upon calcination, resulting in the hollow porous microspheres for the ``top'' product. The nanoscale crystals of the polyanion materials are tightly enwrapped by the highly-conductive framework in the hollow microsphere, resulting in the hierarchical nano-microstructure. The whole formation process is disclosed as a ``bottom-up'' mechanism. Moreover, the biochemistry-directed self-assembly process is confirmed to play a crucial role in the construction of the final architecture. Taking advantage of the well-defined hollow-microsphere architecture, the abundant interior voids and the highly-conductive framework, polyanion materials show favourable sodium-intercalation kinetics. Both materials are capable of high-rate long-term cycling. After five hundred cycles at 20 C and 10 C, Na3V2(PO4)3 and Na3.12Fe2.44(P2O7)2 retain 96.2% and 93.1% of the initial capacity, respectively. Therefore, the biochemistry-directed technique provides a low-cost, highly-efficient and widely applicable strategy to produce high-performance polyanion-based cathodes for sodium ion batteries. Electronic supplementary information (ESI) available: Synthesis, morphology and physical characteristics of reference samples; pore size distribution of the hollow porous microsphere; morphology of the bio-directed Na3.12Fe2.44(P2O7)2 hollow microsphere; comparison of the Rct values for the Na3.12Fe2.44(P2O7)2 composites; TEM images of the cycled materials; lattice parameters and atomic ratio of the samples; calculation process of sodium intercalation coefficients based on the GITT results. See DOI: 10.1039/c6nr00680a
Enhanced autonomic shutdown of Li-ion batteries by polydopamine coated polyethylene microspheres
Baginska, Marta; Blaiszik, Benjamin J.; Rajh, Tijana; ...
2014-07-17
Thermally triggered autonomic shutdown of a Lithium-ion (Li-ion) battery is demonstrated using polydopamine (PDA)-coated polyethylene microspheres applied onto a battery anode. The microspheres are dispersed in a buffered 10 mM dopamine salt solution and the pH is raised to initiate the polymerization and coat the microspheres. Coated microspheres are then mixed with an aqueous binder, applied onto a battery anode surface, dried, and incorporated into Li-ion coin cells. FTIR and Raman spectroscopy are used to verify the presence of the polydopamine on the surface of the microspheres. Scanning electron microscopy is used to examine microsphere surface morphology and resulting anodemore » coating quality. Charge and discharge capacity, as well as impedance, are measured for Li-ion coin cells as a function of microsphere content. Autonomous shutdown is achieved by applying 1.7 mg cm –2 of PDA-coated microspheres to the electrode. Furthermore, the PDA coating significantly reduces the mass of microspheres for effective shutdown compared to our prior work with uncoated microspheres.« less
Ulex europaeus 1 lectin targets microspheres to mouse Peyer's patch M-cells in vivo.
Foster, N; Clark, M A; Jepson, M A; Hirst, B H
1998-03-01
The interaction of latex microspheres with mouse Peyer's patch membranous M-cells was studied in a mouse gut loop model after the microspheres were coated with a variety of agents. Carboxylated microspheres (diameter 0.5 micron) were covalently coated with lectins Ulex europaeus 1, Concanavalin A, Euonymus europaeus and Bandeiraea simplicifolia 1 isolectin-B4, human immunoglobulin A or bovine serum albumin. Of the treatments examined, only Ulex europaeus (UEA1) resulted in significant selective binding of microspheres to M-cells. UEA1-coated microspheres bound to M-cells at a level 100-fold greater than BSA-coated microspheres, but binding to enterocytes was unaffected. Incubation of UEA1-coated microspheres with alpha-L-fucose reduced M-cell binding to a level comparable with BSA-coated microspheres. This indicated that targeting by UEA1 was via a carbohydrate receptor on the M-cell surface. Adherence of UEA1-coated microspheres to M-cells occurred within 10 min of inoculation into mouse gut loops and UEA1-coated microspheres were transported to 10 microns below the apical surface of M-cells within 60 min of inoculation. UEA1-coated microspheres also targeted mouse Peyer's patch M-cells after intragastric administration. These results demonstrated that altering the surface chemistry of carboxylated polystyrene microspheres increased M-cell targeting, suggesting a strategy to enhance delivery of vaccine antigens to the mucosal immune system.
Goldberg, J A; Bradnam, M S; Kerr, D J; McKillop, J H; Bessent, R G; McArdle, C S; Willmott, N; George, W D
1987-12-01
As intra-arterial chemotherapy for liver metastases of colorectal origin becomes accepted, methods of further improving drug delivery to the tumour have been devised. Degradable microspheres have been shown to reduce regional blood flow by transient arteriolar capillary block, thereby improving uptake of a co-administered drug, when injected into the hepatic artery. In our study of five patients, we combined hepatic arterial perfusion scintigraphy (HAPS) and SPECT to assess the localization of approximately 1 X 10(5) labelled microspheres of human serum albumin (99Tcm MSA) in tumour. In addition, in three patients, we assessed the effect of an intra-arterial infusion of the vasoactive agent angiotension II during HAPS. Results were interpreted by comparing transaxial slices with corresponding slices of a tin colloid liver-spleen scan. Two of five patients showed good localization of 99Tcm MSA in tumour without an angiotensin II infusion. Of the three patients receiving angiotensin II, all showed good tumour targetting with the vasoconstrictor compared with only one of these three before its use. Thus, hepatic arterial infusion of angiotensin II greatly improves microsphere localization in tumour in some patients with colorectal liver metastases. This technique may be useful in the assessment of tumour targetting before and during locoregional therapy.
NASA Astrophysics Data System (ADS)
Schroer, M. A.; Gutt, C.; Grübel, G.
2014-07-01
Recently the analysis of scattering patterns by angular cross-correlation analysis (CCA) was introduced to reveal the orientational order in disordered samples with special focus to future applications on x-ray free-electron laser facilities. We apply this CCA approach to ultra-small-angle light-scattering data obtained from two-dimensional monolayers of microspheres. The films were studied in addition by optical microscopy. This combined approach allows to calculate the cross-correlations of the scattering patterns, characterized by the orientational correlation function Ψl(q), as well as to obtain the real-space structure of the monolayers. We show that CCA is sensitive to the orientational order of monolayers formed by the microspheres which are not directly visible from the scattering patterns. By mixing microspheres of different radii the sizes of ordered monolayer domains is reduced. For these samples it is shown that Ψl(q) quantitatively describes the degree of hexagonal order of the two-dimensional films. The experimental CCA results are compared with calculations based on the microscopy images. Both techniques show qualitatively similar features. Differences can be attributed to the wave-front distortion of the laser beam in the experiment. This effect is discussed by investigating the effect of different wave fronts on the cross-correlation analysis results. The so-determined characteristics of the cross-correlation analysis will be also relevant for future x-ray-based studies.
Hooda, Aashima; Nanda, Arun; Jain, Manish; Kumar, Vikash; Rathee, Permender
2012-12-01
The current study involves the development and optimization of their drug entrapment and ex vivo bioadhesion of multiunit chitosan based floating system containing Ranitidine HCl by ionotropic gelation method for gastroretentive delivery. Chitosan being cationic, non-toxic, biocompatible, biodegradable and bioadhesive is frequently used as a material for drug delivery systems and used to transport a drug to an acidic environment where it enhances the transport of polar drugs across epithelial surfaces. The effect of various process variables like drug polymer ratio, concentration of sodium tripolyphosphate and stirring speed on various physiochemical properties like drug entrapment efficiency, particle size and bioadhesion was optimized using central composite design and analyzed using response surface methodology. The observed responses were coincided well with the predicted values given by the optimization technique. The optimized microspheres showed drug entrapment efficiency of 74.73%, particle size 707.26 μm and bioadhesion 71.68% in simulated gastric fluid (pH 1.2) after 8 h with floating lag time 40s. The average size of all the dried microspheres ranged from 608.24 to 720.80 μm. The drug entrapment efficiency of microspheres ranged from 41.67% to 87.58% and bioadhesion ranged from 62% to 86%. Accelerated stability study was performed on optimized formulation as per ICH guidelines and no significant change was found in drug content on storage. Copyright © 2012 Elsevier B.V. All rights reserved.
Hutchens, Thomas C.; Darafsheh, Arash; Fardad, Amir; Antoszyk, Andrew N.; Ying, Howard S.; Astratov, Vasily N.
2012-01-01
Abstract. Ophthalmic surgery may benefit from use of more precise fiber delivery systems during laser surgery. Some current ophthalmic surgical techniques rely on tedious mechanical dissection of tissue layers. In this study, chains of sapphire microspheres integrated into a hollow waveguide distal tip are used for erbium:YAG laser ablation studies in contact mode with ophthalmic tissues, ex vivo. The laser’s short optical penetration depth combined with the small spot diameters achieved with this fiber probe may provide more precise tissue removal. One-, three-, and five-microsphere chain structures were characterized, resulting in FWHM diameters of 67, 32, and 30 μm in air, respectively, with beam profiles comparable to simulations. Single Er:YAG pulses of 0.1 mJ and 75-μs duration produced ablation craters with average diameters of 44, 30, and 17 μm and depths of 26, 10, and 8 μm, for one-, three-, and five-sphere structures, respectively. Microsphere chains produced spatial filtering of the multimode Er:YAG laser beam and fiber, providing spot diameters not otherwise available with conventional fiber systems. Because of the extremely shallow treatment depth, compact focused beam, and contact mode operation, this probe may have potential for use in dissecting epiretinal membranes and other ophthalmic tissues without damaging adjacent retinal tissue. PMID:22734790
Ji, Ye; Xu, Gong Ping; Zhang, Zhi Peng; Xia, Jing Jun; Yan, Jing Long; Pan, Shang Ha
2010-03-01
Autogenous bone grafts are widely used in the repair of bone defects. Growth factors such as bone morphogenetic protein 2 (BMP-2) can induce bone regeneration and enhance bone growth. The combination of an autogenous bone graft and BMP-2 may provide a better osteogenic effect than either treatment alone, but BMP-2 is easily inactivated in body fluid. The objective of this study was to develop a technique that can better preserve the in vivo activity of BMP-2 incorporated in bone grafts. In this study, we first prepared BMP-2/poly(lactic-co-glycolic acid) (PLGA) delayed-release microspheres, and then combined collagen, the delayed-release microspheres, and rat autologous bone particulates to form four groups of composite grafts with different combinations: collagen in group A; collagen combined with bone particulates in group B; collagen combined with BMP-2/PLGA delayed-release microspheres in group C; and collagen combined with both bone particulates and BMP-2/PLGA delayed-release microspheres in group D. The four groups of composite grafts were implanted into the gluteus maximus pockets in rats. The ectopic osteogenesis and ALP level in group D (experimental group) were compared with those in groups A, B, and C (control groups) to study whether it had higher osteogenic capability. Results showed that the composite graft design increased the utility of BMP-2 and reduced the required dose of BMP-2 and volume of autologous bone. The selection of bone particulate diameter had an impact on the osteogenetic potential of bone grafts. Collagen prevented the occurrence of aseptic inflammation and improved the osteoinductivity of BMP-2. These results showed that this composite graft design is effective and feasible for use in bone repair.
Effect of a dual inlet channel on cell loading in microfluidics.
Yun, Hoyoung; Kim, Kisoo; Lee, Won Gu
2014-11-01
Unwanted sedimentation and attachment of a number of cells onto the bottom channel often occur on relatively large-scale inlets of conventional microfluidic channels as a result of gravity and fluid shear. Phenomena such as sedimentation have become recognized problems that can be overcome by performing microfluidic experiments properly, such as by calculating a meaningful output efficiency with respect to real input. Here, we present a dual-inlet design method for reducing cell loss at the inlet of channels by adding a new " upstream inlet " to a single main inlet design. The simple addition of an upstream inlet can create a vertically layered sheath flow prior to the main inlet for cell loading. The bottom layer flow plays a critical role in preventing the cells from attaching to the bottom of the channel entrance, resulting in a low possibility of cell sedimentation at the main channel entrance. To provide proof-of-concept validation, we applied our design to a microfabricated flow cytometer system (μFCS) and compared the cell counting efficiency of the proposed μFCS with that of the previous single-inlet μFCS and conventional FCS. We used human white blood cells and fluorescent microspheres to quantitatively evaluate the rate of cell sedimentation in the main inlet and to measure fluorescence sensitivity at the detection zone of the flow cytometer microchip. Generating a sheath flow as the bottom layer was meaningfully used to reduce the depth of field as well as the relative deviation of targets in the z-direction (compared to the x-y flow plane), leading to an increased counting sensitivity of fluorescent detection signals. Counting results using fluorescent microspheres showed both a 40% reduction in the rate of sedimentation and a 2-fold higher sensitivity in comparison with the single-inlet μFCS. The results of CD4(+) T-cell counting also showed that the proposed design results in a 25% decrease in the rate of cell sedimentation and a 28% increase in sensitivity when compared to the single-inlet μFCS. This method is simple and easy to use in design, yet requires no additional time or cost in fabrication. Furthermore, we expect that this approach could potentially be helpful for calculating exact cell loading and counting efficiency for a small input number of cells, such as primary cells and rare cells, in microfluidic channel applications.
NASA Astrophysics Data System (ADS)
Jiang, J. F.; Zhao, Q.; Lin, M. Q.; Wang, Y. F.; Dang, S. M.; Sun, F. F.
2015-12-01
Terpolymeric microspheres were synthesized by the inverse suspension polymerization of functional monomers including AMPS, NVP, and AM. The morphology and size of the obtained microspheres were measured by scanning electron microscopy (SEM) and optical microscopy. Furthermore, the swelling performances of the obtained microspheres were measured with alaser particle analyzer (LPA), and the thermal stability of the microspheres obtained was measured by differential thermal analysis (DSC-TG) and high temperature experiments involving microsphere/water dispersion. The results revealed that the extreme value of the microsphere size distribution decreased from 280 μm to 20 μm as the stirring rate increased from 175 rpm to 500 rpm. At temperatures below 25°C, the maximum achieved swelling ratio of the microspheres was 21, and the thermal stability of the terpolymer microspheres was significantly higher than that of the dipolymer microspheres. The terpolymer/water dispersions were kept at 120°C for 19d before any damage was observed.
Microradiographic microsphere manipulator
Singleton, R.M.
A method and apparatus is disclosed for radiographic characterization of small hollow spherical members (microspheres), constructed of either optically transparent or opaque materials. The apparatus involves a microsphere manipulator which holds a batch of microspheres between two parallel thin plastic films for contact microradiographic characterization or projection microradiography thereof. One plastic film is translated relative to and parallel to the other to roll the microspheres through any desired angle to allow different views of the microspheres.
Microradiographic microsphere manipulator
Singleton, Russell M.
1980-01-01
A method and apparatus for radiographic characterization of small hollow spherical members (microspheres), constructed of either optically transparent or opaque materials. The apparatus involves a microsphere manipulator which holds a batch of microspheres between two parallel thin plastic films for contact microradiographic characterization or projection microradiography thereof. One plastic film is translated to relative to and parallel to the other to roll the microspheres through any desired angle to allow different views of the microspheres.
Lin, Guanquan; Chen, Huayao; Zhou, Hongjun; Zhou, Xinhua; Xu, Hua
2018-01-01
Using butyl methacrylate (BMA) and styrene (St) as monomers and divinylbenzene (DVB) as a crosslinking agent, P(St-BMA) microspheres were prepared by suspension polymerization. Tea tree oil (TTO) microspheres were prepared by adsorbing TTO on P(St-BMA) microspheres. The structure and surface morphology of P(St-BMA) microspheres and TTO microspheres were characterized by Fourier transformed infrared spectroscopy (FTIR), optical microscopy, and Thermogravimetric analysis (TGA). In doing so, the structural effect of P(St-BMA) microspheres on oil absorption and sustained release properties could be investigated. The results show that the surface of the P(St-BMA) microspheres in the process of TTO microsphere formation changed from initially concave to convex. The TTO microspheres significantly improved the stability of TTO, which was found to completely decompose as the temperature of the TTO increased from about 110 °C to 150 °C. The oil absorption behavior, which was up to 3.85 g/g, could be controlled by adjusting the monomer ratio and the amount of crosslinking agent. Based on Fickian diffusion, the sustained release behavior of TTO microspheres was consistent with the Korsmeyer-Pappas kinetic model. After 13 h of natural release, the anti-bacterial effect of the TTO microspheres was found to be significantly improved compared to TTO. PMID:29723967
Preparation and drug controlled release of porous octyl-dextran microspheres.
Hou, Xin; Liu, Yanfei
2015-01-01
In this work, porous octyl-dextran microspheres with excellent properties were prepared by two steps. Firstly, dextran microspheres were synthesized by reversed-phase suspension polymerization. Secondly, octyl-dextran microspheres were prepared by the reaction between dextran microspheres and ethylhexyl glycidyl ether and freezing-drying method. Porous structure of microspheres was formed through the interaction between octyl groups and organic solvents. The structure, morphology, dry density, porosity and equilibrium water content of porous octyl-dextran microspheres were systematically investigated. The octyl content affected the properties of microspheres. The results showed that the dry density of microspheres decreased from 2.35 to 1.21 g/ml, porosity increased from 80.68 to 95.05% with the octyl content increasing from 0.49 to 2.28 mmol/g. Meanwhile, the equilibrium water content presented a peak value (90.18%) when the octyl content was 2.25 mmol/g. Octyl-dextran microspheres showed high capacity. Naturally drug carriers play an important role in drug-delivery systems for their biodegradability, wide raw materials sources and nontoxicity. Doxorubicin (DOX) was used as a drug model to examine the drug-loading capacity of porous octyl-dextran microspheres. The drug-loading efficiency increased with the increase in microspheres/drug ratio, while the encapsulation efficiency decreased. When microspheres/drug mass ratio was 4/1, the drug-loading efficiency and encapsulation efficiency were 10.20 and 51.00%, respectively. The release rate of DOX increased as drug content and porosity increased. In conclusion, porous octyl-dextran microspheres were synthesized successfully and have the potential to serve as an effective delivery system in drug controlled release.
Fluorescence-Doped Particles for Simultaneous Temperature and Velocity Imaging
NASA Technical Reports Server (NTRS)
Danehy, Paul M.; Tiemsin, Pacita I.; Wohl, Chrostopher J.; Verkamp, Max; Lowe, T.; Maisto, P.; Byun, G.; Simpson, R.
2012-01-01
Polystyrene latex microspheres (PSLs) have been used for particle image velocimetry (PIV) and laser Doppler velocimetry (LDV) measurements for several decades. With advances in laser technologies, instrumentation, and data processing, the capability to collect more information about fluid flow beyond velocity is possible using new seed materials. To provide additional measurement capability, PSLs were synthesized with temperature-sensitive fluorescent dyes incorporated within the particle. These multifunctional PSLs would have the greatest impact if they could be used in large scale facilities with minimal modification to the facilities or the existing instrumentation. Consequently, several potential dyes were identified that were amenable to existing laser systems currently utilized in wind tunnels at NASA Langley Research Center as well as other wind and fluid (water) tunnels. PSLs incorporated with Rhodamine B, dichlorofluorescein (DCF, also known as fluorescein 548 or fluorescein 27) and other dyes were synthesized and characterized for morphology and spectral properties. The resulting particles were demonstrated to exhibit fluorescent emission, which would enable determination of both fluid velocity and temperature. They also would allow near-wall velocity measurements whereas laser scatter from surfaces currently prevents near-wall measurements using undoped seed materials. Preliminary results in a wind tunnel facility located at Virginia Polytechnic Institute and State University (Virginia Tech) have verified fluorescent signal detection and temperature sensitivity of fluorophore-doped PSLs.
Zhang, An-yang; Fan, Tian-yuan
2009-12-18
To investigate the preparation, optimization and in vitro properties of riboflavin sodium phosphate floating microspheres. The floating microspheres composed of riboflavin sodium phosphate and calcium alginate were prepared using ion gelatin-oven drying method. The properties of the microspheres were investigated, including the buoyancy, release, appearance and entrapment efficiency. The formulation was optimized by response surface methodology (RSM). The optimized microspheres were round. The entrapment efficiency was 57.49%. All the microspheres could float on the artificial gastric juice over 8 hours. The release of the drug from the microspheres complied with Fick's diffusion.
Wang, Ke; Wang, Yinjing; Zhao, Xu; Li, Yi; Yang, Tao; Zhang, Xue; Wu, Xiaoguang
2017-06-01
Hollow carbonated hydroxyapatite (HCHAp) microspheres as simvastatin (SV) sustained-release vehicles were fabricated through a novel and simple one-step biomimetic strategy. Firstly, hollow CaCO 3 microspheres were precipitated through the reaction of CaCl 2 with Na 2 CO 3 in the presence of aspartic acid and sodium dodecyl sulfate. Then, the as-prepared hollow CaCO 3 microspheres were transformed into HCHAp microspheres with a controlled anion-exchange method. The HCHAp microspheres were 3-5μm with a shell thickness of 0.5-1μm and were constructed of short needle nanoparticles. The HCHAp microspheres were then loaded with SV, exhibiting excellent drug-loading capacity and sustained release properties. These results present a new material synthesis strategy for HCHAp microspheres and suggest that the as-prepared HCHAp microspheres are promising for applications in drug delivery. Copyright © 2017 Elsevier B.V. All rights reserved.
Berchane, N S; Carson, K H; Rice-Ficht, A C; Andrews, M J
2007-06-07
The need to tailor release rate profiles from polymeric microspheres is a significant problem. Microsphere size, which has a significant effect on drug release rate, can potentially be varied to design a controlled drug delivery system with desired release profile. In this work the effects of microspheres mean diameter, polydispersity, and polymer degradation on drug release rate from poly(lactide-co-glycolide) (PLG) microspheres are described. Piroxicam containing PLG microspheres were fabricated at 20% loading, and at three different impeller speeds. A portion of the microspheres was then sieved giving five different size distributions. In vitro release kinetics were determined for each preparation. Based on these experimental results, a suitable mathematical theory has been developed that incorporates the effect of microsphere size distribution and polymer degradation on drug release. We show from in vitro release experiments that microsphere size has a significant effect on drug release rate. The initial release rate decreased with an increase in microsphere size. In addition, the release profile changed from first order to concave-upward (sigmoidal) as the microsphere size was increased. The mathematical model gave a good fit to the experimental release data. For highly polydisperse populations (polydispersity parameter b<3), incorporating the microsphere size distribution into the mathematical model gave a better fit to the experimental results than using the representative mean diameter. The validated mathematical model can be used to predict small-molecule drug release from PLG microsphere populations.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zhou, Xiaoliang; Yan, Zhengguang, E-mail: yanzg2004@gmail.com; Han, Xiaodong, E-mail: xdhan@bjut.edu.cn
2014-02-01
Graphical abstract: In situ growth of copper nanoparticles from hydrothermal copper-containing carbonaceous microspheres was induced by annealing or electron beam irradiation. Obtained micro-nano carbon/copper composite microspheres show electrochemical glucose sensing properties. - Highlights: • We synthesized carbonaceous microspheres containing non-nanoparicle copper species through a hydrothermal route. • By annealing or electron beam irradiation, copper nanoparticles would form from the carbonaceous microspheres in situ. • By controlling the annealing temperature, particle size of copper could be controlled in the range of 50–500 nm. • The annealed carbon/copper hierarchical composite microspheres were used to fabricate an electrochemical glucose sensor. - Abstract: Inmore » situ growth of copper nanocrystals from carbon/copper microspheres was observed in a well-controlled annealing or an electron beam irradiation process. Carbonaceous microspheres containing copper species with a smooth appearance were yielded by a hydrothermal synthesis using copper nitrate and ascorbic acid as reactants. When annealing the carbonaceous microspheres under inert atmosphere, copper nanoparticles were formed on carbon microspheres and the copper particle sizes can be increased to a range of 50–500 nm by altering the heating temperature. Similarly, in situ formation of copper nanocrystals from these carbonaceous microspheres was observed on the hydrothermal product carbonaceous microspheres with electron beam irradiation in a vacuum transmission electron microscopy chamber. The carbon/copper composite microspheres obtained through annealing were used to modify a glassy carbon electrode and tested as an electrochemical glucose sensor.« less
Contact microspherical nanoscopy: from fundamentals to biomedical applications
NASA Astrophysics Data System (ADS)
Astratov, V. N.; Maslov, A. V.; Brettin, A.; Blanchette, K. F.; Nesmelov, Y. E.; Limberopoulos, N. I.; Walker, D. E.; Urbas, A. M.
2017-02-01
The mechanisms of super-resolution imaging by contact microspherical or microcylindrical nanoscopy remain an enigmatic question since these lenses neither have an ability to amplify the near-fields like in the case of far-field superlens, nor they have a hyperbolic dispersion similar to hyperlenses. In this work, we present results along two lines. First, we performed numerical modeling of super-resolution properties of two-dimensional (2-D) circular lens in the limit of wavelength-scale diameters, λ <= D <= 2λ, and relatively high indices of refraction, n=2. Our preliminary results on imaging point dipoles indicate that the resolution is generally close to λ/4 however on resonance with whispering gallery modes it may be slightly higher. Second, experimentally, we used actin protein filaments for the resolution quantification in microspherical nanoscopy. The critical feature of our approach is based on using arrayed cladding layer with strong localized surface plasmon resonances. This layer is used for enhancing plasmonic near-field illumination of our objects. In combination with the magnification of virtual image, this technique resulted in the lateral resolution of actin protein filaments on the order of λ/7.
Wang, Hongbin; Liu, Lijia; Wang, Runwei; Yan, Xiao; Wang, Ziqi; Hu, Jiangtao; Chen, Haibiao; Jiang, Shang; Ni, Ling; Qiu, Hailong; Tang, Haitong; Wei, Yingjin; Zhang, Zongtao; Qiu, Shilun; Pan, Feng
2018-05-18
LiFePO 4 @C/reduced graphene oxide (rGO) hierarchical microspheres with superior electrochemical activity and a high tap density were first synthesized by using a Fe 3+ -based single inorganic precursor (LiFePO 4 OH@RF/GO; RF=resorcinol-formaldehyde, GO=graphene oxide) obtained from a template-free self-assembly synthesis followed by direct calcination. The synthetic process requires no physical mixing step. The phase transformation pathway from tavorite LiFePO 4 OH to olivine LiFePO 4 upon calcination was determined by means of the in situ high-temperature XRD technique. Benefitting from the unique structure of the material, these microspheres can be densely packed together, giving a high tap density of 1.3 g cm -3 , and simultaneously, defectless LiFePO 4 primary nanocrystals modified with a highly conductive surface carbon layer and ultrathin rGO provide good electronic and ionic kinetics for fast electron/Li + ion transport. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
Glass microspheres for medical applications
NASA Astrophysics Data System (ADS)
Conzone, Samuel David
Radioactive dysprosium lithium borate glass microspheres have been developed as biodegradable radiation delivery vehicles for the radiation synovectomy treatment of rheumatoid arthritis. Once injected into a diseased joint, the microspheres deliver a potent dose of radiation to the diseased tissue, while a non-uniform chemical reaction converts the glass into an amorphous, porous, hydrated dysprosium phosphate reaction product. The non-radioactive, lithium-borate component is dissolved from the glass (up to 94% weight loss), while the radioactive 165Dy reacts with phosphate anions in the body fluids, and becomes "chemically" trapped in a solid, dysprosium phosphate reaction product that has the same size as the un-reacted glass microsphere. Ethylene diamine tetraacetate (EDTA) chelation therapy can be used to dissolve the dysprosium phosphate reaction product after the radiation delivery has subsided. The dysprosium phosphate reaction product, which formed in vivo in the joint of a Sprague-Dawley rat, was dissolved by EDTA chelation therapy in <1 week, without causing any detectable joint damage. The combination of dysprosium lithium borate glass microspheres and EDTA chelation therapy provides an unique "tool" for the medical community, which can deliver a large dose (>100 Gy) of localized beta radiation to a treatment site within the body, followed by complete biodegradability. The non-uniform reaction process is a desirable characteristic for a biodegradable radiation delivery vehicle, but it is also a novel material synthesis technique that can convert a glass to a highly porous materials with widely varying chemical composition by simple, low-temperature, glass/solution reaction. The reaction product formed by nonuniform reaction occupies the same volume as the un-reacted glass, and after drying for 1 h at 300°C, has a specific surface area of ≈200 m2/g, a pore size of ≈30 nm, and a nominal crushing strength of ≈10 MPa. Finally, rhenium glass microspheres, composed of micron-sized, metallic rhenium particles dispersed within a magnesium alumino borate glass matrix were produced by sintering ReO2 powder and glass frit at 1050°C. A 50 mg injection of radioactive rhenium glass microspheres containing 3.7 GBq of 186Re and 8.5 GBq of 188Re could be used to deliver a 100 Gy dose to a cancerous tumor, while limiting the total body dose caused by rhenium dissolution to approximately 1 mGy.
Wang, Weina; Ma, Ruiyang; Wu, Qiuhua; Wang, Chun; Wang, Zhi
2013-06-07
In this paper, a magnetic microsphere-confined graphene adsorbent (Fe3O4@SiO2-G) was fabricated and used for the extraction of five polycyclic aromatic hydrocarbons (fluorene, anthracene, phenanthrene, fluoranthene and pyrene) from environmental water samples prior to high performance liquid chromatography with fluorescence detection. The Fe3O4@SiO2-G was characterized by various instrumental methods. Various experimental parameters that could affect the extraction efficiencies, such as the amount of Fe3O4@SiO2-G, the pH and ionic strength of sample solution, the extraction time and the desorption conditions, were investigated. Due to the high surface area and excellent adsorption capacity of the Fe3O4@SiO2-G, satisfactory extraction can be achieved with only 15mg of the adsorbent per 250mL solution and 5min extraction. Under the optimum conditions, a linear response was observed in the concentration range of 5-1500ngL(-1) for fluorene, 2.5-1500ngL(-1) for anthracene and 15-1500ngL(-1) for phenanthrene, fluoranthene and pyrene, with the correlation coefficients (r) ranging from 0.9897 to 0.9961. The limits of detection (S/N=3) of the method were between 0.5 and 5.0ngL(-1). The relative standard deviations (RSDs) were less than 5.6%. The recoveries of the method were in the range between 83.2% and 108.2%. The results indicated that this graphene-based magnetic nanocomposite had a great adsorptive ability toward the five polycyclic aromatic hydrocarbons from environmental water samples. Copyright © 2013 Elsevier B.V. All rights reserved.
Dumonceaux, Tim J.; Green, Margaret; Hammond, Christine; Perez, Edel; Olivier, Chrystel
2014-01-01
Phytoplasmas (‘Candidatus Phytoplasma’ spp.) are insect-vectored bacteria that infect a wide variety of plants, including many agriculturally important species. The infections can cause devastating yield losses by inducing morphological changes that dramatically alter inflorescence development. Detection of phytoplasma infection typically utilizes sequences located within the 16S–23S rRNA-encoding locus, and these sequences are necessary for strain identification by currently accepted standards for phytoplasma classification. However, these methods can generate PCR products >1400 bp that are less divergent in sequence than protein-encoding genes, limiting strain resolution in certain cases. We describe a method for accessing the chaperonin-60 (cpn60) gene sequence from a diverse array of ‘Ca.Phytoplasma’ spp. Two degenerate primer sets were designed based on the known sequence diversity of cpn60 from ‘Ca.Phytoplasma’ spp. and used to amplify cpn60 gene fragments from various reference samples and infected plant tissues. Forty three cpn60 sequences were thereby determined. The cpn60 PCR-gel electrophoresis method was highly sensitive compared to 16S-23S-targeted PCR-gel electrophoresis. The topology of a phylogenetic tree generated using cpn60 sequences was congruent with that reported for 16S rRNA-encoding genes. The cpn60 sequences were used to design a hybridization array using oligonucleotide-coupled fluorescent microspheres, providing rapid diagnosis and typing of phytoplasma infections. The oligonucleotide-coupled fluorescent microsphere assay revealed samples that were infected simultaneously with two subtypes of phytoplasma. These tools were applied to show that two host plants, Brassica napus and Camelina sativa, displayed different phytoplasma infection patterns. PMID:25551224
Francis, Andrew W.; Kagemann, Larry; Wollstein, Gadi; Ishikawa, Hiroshi; Folz, Steven; Overby, Darryl R.; Sigal, Ian A.; Wang, Bo; Schuman, Joel S.
2012-01-01
Purpose. To describe morphometric details of the human aqueous humor (AH) outflow microvasculature visualized with 360-degree virtual castings during active AH outflow in cadaver eyes and to compare these structures with corrosion casting studies. Methods. The conventional AH outflow pathways of donor eyes (n = 7) and eyes in vivo (n = 3) were imaged with spectral-domain optical coherence tomography (SD-OCT) and wide-bandwidth superluminescent diode array during active AH outflow. Digital image contrast was adjusted to isolate AH microvasculature, and images were viewed in a 3D viewer. Additional eyes (n = 3) were perfused with mock AH containing fluorescent tracer microspheres to compare microvasculature patterns. Results. Observations revealed components of the conventional outflow pathway from Schlemm's canal (SC) to the superficial intrascleral venous plexus (ISVP). The superficial ISVP in both our study and corrosion casts were composed of interconnected venules (10–50 μm) forming a hexagonal meshwork. Larger radial arcades (50–100 μm) drained the region nearest SC and converged with larger tortuous vessels (>100 μm). A 360-degree virtual casting closely approximated corrosion casting studies. Tracer studies corroborated our findings. Tracer decorated several larger vessels (50–100 μm) extending posteriorly from the limbus in both raw and contrast-enhanced fluorescence images. Smaller tracer-labeled vessels (30–40 μm) were seen branching between larger vessels and exhibited a similar hexagonal network pattern. Conclusions. SD-OCT is capable of detailed morphometric analysis of the conventional outflow pathway in vivo or ex vivo with details comparable to corrosion casting techniques. PMID:22499987
Viswanathan, N B; Thomas, P A; Pandit, J K; Kulkarni, M G; Mashelkar, R A
1999-03-08
Emulsification-solvent removal methods have been widely used for encapsulating bioactive macromolecules like proteins and polypeptides in biodegradable polymers. We report, a (water-in-oil)-in-oil emulsion technique wherein proteins and polypeptides differing in molecular weight and shape were encapsulated in polymers of current biomedical interest. When an oil was used as the processing medium in combination with a carefully selected mixed solvent system such that a stable (w/o1/o2 emulsion is formed and solvents are removed by a combination of extraction and evaporation, the entrapment efficiency was high and the product nonporous. The entrapment efficiency of globular proteins exceeded 90% while that of fibrous proteins was around 70%. Fracture studies revealed that the polymer matrix was dense. The mechanism of entrapment involved solvent-induced precipitation of the protein as the microspheres were being formed. The principle of the method will find use in preparation of non-porous polymer microparticles with reduced burst effect.
Preparation and release characteristics of polymer-coated and blended alginate microspheres.
Lee, D W; Hwang, S J; Park, J B; Park, H J
2003-01-01
To prevent a rapid drug release from alginate microspheres in simulated intestinal media, alginate microspheres were coated or blended with polymers. Three polymers were selected and evaluated such as HPMC, Eudragit RS 30D and chitosan, as both coating materials and additive polymers for controlling the drug release. This study focused on the release characteristics of polymer-coated and blended alginate microspheres, varying the type of polymer and its concentration. The alginate microspheres were prepared by dropping the mixture of drug and sodium alginate into CaCl(2) solution using a spray-gun. Polymer-coated microspheres were prepared by adding alginate microspheres into polymer solution with mild stirring. Polymer-blended microspheres were prepared by dropping the mixture of drug, sodium alginate and additive polymer with plasticizer into CaCl(2) solution. In vitro release test was carried out to investigate the release profiles in 500 ml of phosphate buffered saline (PBS, pH 7.4). As the amount of polymer in sodium alginate or coating solution increase, the drug release generally decreased. HPMC-blended microspheres swelled but withstood the disintegration, showing an ideal linear release profiles. Chitosan-coated microspheres showed smooth and round surface and extended the release of drug. In comparison with chitosan-coated microspheres, HPMC-blended alginate microspheres can be easily made and used for controlled drug delivery systems due to convenient process and controlled drug release.
Experimental study on microsphere assisted nanoscope in non-contact mode
NASA Astrophysics Data System (ADS)
Ling, Jinzhong; Li, Dancui; Liu, Xin; Wang, Xiaorui
2018-07-01
Microsphere assisted nanoscope was proposed in existing literatures to capture super-resolution images of the nano-structures beneath the microsphere attached on sample surface. In this paper, a microsphere assisted nanoscope working in non-contact mode is designed and demonstrated, in which the microsphere is controlled with a gap separated to sample surface. With a gap, the microsphere is moved in parallel to sample surface non-invasively, so as to observe all the areas of interest. Furthermore, the influence of gap size on image resolution is studied experimentally. Only when the microsphere is close enough to the sample surface, super-resolution image could be obtained. Generally, the resolution decreases when the gap increases as the contribution of evanescent wave disappears. To keep an appropriate gap size, a quantitative method is implemented to estimate the gap variation by observing Newton's rings around the microsphere, serving as a real-time feedback for tuning the gap size. With a constant gap, large-area image with high resolution can be obtained during microsphere scanning. Our study of non-contact mode makes the microsphere assisted nanoscope more practicable and easier to implement.
Microsphere-assisted super-resolution imaging with enlarged numerical aperture by semi-immersion
NASA Astrophysics Data System (ADS)
Wang, Fengge; Yang, Songlin; Ma, Huifeng; Shen, Ping; Wei, Nan; Wang, Meng; Xia, Yang; Deng, Yun; Ye, Yong-Hong
2018-01-01
Microsphere-assisted imaging is an extraordinary simple technology that can obtain optical super-resolution under white-light illumination. Here, we introduce a method to improve the resolution of a microsphere lens by increasing its numerical aperture. In our proposed structure, BaTiO3 glass (BTG) microsphere lenses are semi-immersed in a S1805 layer with a refractive index of 1.65, and then, the semi-immersed microspheres are fully embedded in an elastomer with an index of 1.4. We experimentally demonstrate that this structure, in combination with a conventional optical microscope, can clearly resolve a two-dimensional 200-nm-diameter hexagonally close-packed (hcp) silica microsphere array. On the contrary, the widely used structure where BTG microsphere lenses are fully immersed in a liquid or elastomer cannot even resolve a 250-nm-diameter hcp silica microsphere array. The improvement in resolution through the proposed structure is due to an increase in the effective numerical aperture by semi-immersing BTG microsphere lenses in a high-refractive-index S1805 layer. Our results will inform on the design of microsphere-based high-resolution imaging systems.
NASA Astrophysics Data System (ADS)
Aceves, A.; Pierson, J. J.; Millette, N.
2016-02-01
Mixotrophic plankton are capable of obtaining their energy through photosynthesis and phagocytosis, and have been observed to be common among marine and freshwater dinoflagellates. The role of mixotrophic dinoflagellates in the `microbial loop' has received little attention. Organisms that were only thought to introduce new carbon into the loop through photosynthesis may also consume fixed carbon by ingesting bacteria, making the `microbial loop' more complex that originally conceived. The nanodinoflagellate Heterocapsa rotundata was cultured under various light and nutrient regimes to investigate the role of phototrophy and phagotrophy during winter conditions in the Chesapeake Bay. We quantified grazing rates of H. rotundata on bacteria using two feeding methods, ingestion of polycarbonate microspheres and prey removal experiments. Ingestion of fluorescent microspheres by H. rotundata revealed their ability to phagocytize particles. Using flow cytometry we calculated grazing rates of H. rotundata on bacteria under various light intensities and ammonium concentrations and found that H. rotundata increased phagotrophy at lower light intensities and ammonium was positively correlated with the grazing rates of H. rotundata. We conclude that H. rotundata uses mixotrophy as a primary source for obtaining carbon during the winter when there is limited light and lower temperatures.
Yang, Liangbao; Han, Jun; Luo, Tao; Li, Minqiang; Huang, Jiarui; Meng, Fanli; Liu, Jinhuai
2009-01-05
Almost monodisperse ZnS microspheres have been synthesized on a large scale by a hydrothermal route, in which tungstosilicate acid (TSA) was used as a soft template. By controlling the reaction conditions, such as reaction temperature, pH value of the solutions, and the reaction medium, almost monodisperse microspheres can be synthesized. The structure of these microspheres is sensitive to the reaction conditions. The growth mechanism of these nearly monodisperse microspheres was examined. Oxygen sensing is realized from ZnS microspheres. The current through the ZnS microspheres under UV illumination increases as the oxygen concentration decreases.
Meng, Boyu; Li, Ling; Hua, Su; Wang, Qingsong; Liu, Chunhui; Xu, Xiangyang; Yin, Xiaojin
2010-09-15
The incomplete release of Endostar from PLGA microspheres was observed in our previous study. In the present study, we focused on the effect of medium-chain triglycerides (MCT) on the in vitro/in vivo release behavior of Endostar encapsulated PLGA microspheres, which were prepared by a water-in-oil-in-water (W/O/W) double-emulsion method with or without MCT. The in vitro accumulated release of Endostar from microspheres co-encapsulated with 30% MCT was found to be 79.04% after a 30-day incubation period in PBS (pH 7.4) at 37 degrees C. However, the accumulated release of Endostar from MCT-free microspheres was found to be only 32.22%. Pouches containing Endostar encapsulated PLGA microspheres were implanted subcutaneously in rats. The effect of MCT on the in vivo release showed a similar trend to the in vitro release. After 30 days, only 9.87% of the total encapsulated Endostar was retained in microspheres co-encapsulated with 30% MCT, while 42.25% of Endostar was retained in MCT-free microspheres. The co-encapsulation of MCT provided the microspheres with a porous surface, which significantly improved the in vitro/in vivo release of Endostar from PLGA microspheres. In addition, in vitro experiments showed that MCT co-encapsulated PLGA microspheres had more inter-connected pores, faster degradation of PLGA, and faster swelling of microspheres, which helped to explain the mechanism of the effect of MCT on improving the release of Endostar from PLGA microspheres. Copyright 2010 Elsevier B.V. All rights reserved.
Alcalá-Alcalá, Sergio; Benítez-Cardoza, Claudia G; Lima-Muñoz, Enrique J; Piñón-Segundo, Elizabeth; Quintanar-Guerrero, David
2015-07-15
This work presents an evaluation of the adsorption/infiltration process in relation to the loading of a model protein, α-amylase, into an assembled biodegradable polymeric system, free of organic solvents and made up of poly(D,L-lactide-co-glycolide) acid (PLGA). Systems were assembled in a friendly aqueous medium by adsorbing and infiltrating polymeric nanoparticles into porous microspheres. These assembled systems are able to load therapeutic amounts of the drug through adsorption of the protein onto the large surface area characteristic of polymeric nanoparticles. The subsequent infiltration of nanoparticles adsorbed with the protein into porous microspheres enabled the controlled release of the protein as a function of the amount of infiltrated nanoparticles, since the surface area available on the porous structure is saturated at different levels, thus modifying the protein release rate. Findings were confirmed by both the BET technique (N2 isotherms) and in vitro release studies. During the adsorption process, the pH of the medium plays an important role by creating an environment that favors adsorption between the surfaces of the micro- and nano-structures and the protein. Finally, assays of α-amylase activity using 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNP-G3) as the substrate and the circular dichroism technique confirmed that when this new approach was used no conformational changes were observed in the protein after release. Copyright © 2015 Elsevier B.V. All rights reserved.
NASA Astrophysics Data System (ADS)
Seok Lyoo, Won; Wook Cha, Jin; Young Kwak, Kun; Jae Lee, Young; Yong Jeon, Han; Sik Chung, Yong; Kyun Noh, Seok
2010-06-01
To prepare Poly(vinyl pivalate/vinyl acetate) [P(VPi/VAc)] microspheres with radiopacity, the suspension copolymerization approach in the presence of aqueous radiopaque nanoparticles was used. After, The P(VPi/VAc) microspheres with radiopacity were saponified in heterogeneous system, and then P(VPi/VAc) microspheres without aggregates were converted to s-PVA/P(VPi/VAc) microspheres of skin/core structure through the heterogeneous surface saponification. Radiopacity of microspheres was confirmed with Computed tomography (CT).
Mesoporous metal oxide microsphere electrode compositions and their methods of making
DOE Office of Scientific and Technical Information (OSTI.GOV)
Parans Paranthaman, Mariappan; Bi, Zhonghe; Bridges, Craig A.
Compositions and methods of making are provided for treated mesoporous metal oxide microspheres electrodes. The compositions include microspheres with an average diameter between about 200 nanometers and about 10 micrometers and mesopores on the surface and interior of the microspheres. The methods of making include forming a mesoporous metal oxide microsphere composition and treating the mesoporous metal oxide microspheres by at least annealing in a reducing atmosphere, doping with an aliovalent element, and coating with a coating composition.
Production of hollow aerogel microspheres
Upadhye, Ravindra S.; Henning, Sten A.
1993-01-01
A method is described for making hollow aerogel microspheres of 800-1200 .mu. diameter and 100-300 .mu. wall thickness by forming hollow alcogel microspheres during the sol/gel process in a catalytic atmosphere and capturing them on a foam surface containing catalyst. Supercritical drying of the formed hollow alcogel microspheres yields hollow aerogel microspheres which are suitable for ICF targets.
5-Fluorouracil:carnauba wax microspheres for chemoembolization: an in vitro evaluation.
Benita, S; Zouai, O; Benoit, J P
1986-09-01
5-Fluorouracil:carnauba wax microspheres were prepared using a meltable dispersion process with the aid of a surfactant as a wetting agent. It was noted that only hydrophilic surfactants were able to wet the 5-fluorouracil and substantially increased its content in the microspheres. No marked effect was observed in the particle size distribution of the solid microspheres as a function of the nature of the surfactant. Increasing the stirring rate in the preparation process decreased, first, the mean droplet size of the emulsified melted dispersion in the vehicle during the heating process, and, consequently, the mean particle size of the solidified microspheres during the cooling process. 5-Fluorouracil cumulative release from the microspheres followed first-order kinetics, as shown by nonlinear regression analysis. Although the kinetic results were not indicative of the true release mechanism from a single microsphere, it was believed that 5-fluorouracil release from the microspheres was probably governed by a dissolution process, rather than by a leaching process through the carnauba wax microspheres.
Topological Transformation of Defects in Nematic Liquid Crystals
NASA Astrophysics Data System (ADS)
Pagel, Zachary; Atherton, Timothy; Guasto, Jeffrey; Cebe, Peggy
A topological transformation around silica microsphere inclusions in nematic liquid crystal cells (LCC) is experimentally studied. Silica microspheres are coated to induce homeotropic LC anchoring to the spheres. Parallel rub directions of the alignment polymer during LCC construction create a splay wall that traps the microspheres. Application of an out-of-plane electric field then permits a transformation of hedgehog defects, reversing the orientation of the defect around microspheres. The transformation controllably reverses the microsphere's direction of travel during AC electrophoresis due to defect-dependent velocity anisotropy. A similar transformation is studied on chains of microspheres with hedgehog defects, where the defect orientation is reversed on the entire chain. Polarized and confocal microscopies are used to study the defect structures. Results contribute to recent developments in microsphere electrokinetics in nematic LCs, as the transformation adds an additional degree of control in the electrophoretic motion of microspheres and chains of microspheres with dipolar defects. The author thanks NSF Grant DMR-1608126 for funding reseearch and Tufts University for funding travel.
Methods for determination of optic nerve blood flow.
Glazer, L. C.
1988-01-01
A variety of studies have been conducted over the past two decades to determine if decreased optic nerve blood flow has a role in the etiology of glaucomatous nerve damage. Five basic methods have been employed in examining blood flow. Invasive studies, utilizing electrodes placed in the optic nerve head, represent one of the first attempts to measure blood flow. More recently, the methodologies have included axoplasmic flow analysis, microspheres, radioactive tracers such as iodoantipyrine, and laser doppler measurements. The results of these studies are inconclusive and frequently contradictory. When the studies are grouped by methodology, only the iodoantipyrine data are consistent. While each of the experimental techniques has limitations, iodoantipyrine appears to have better resolution than either invasive studies or microspheres. PMID:3284212
NASA Astrophysics Data System (ADS)
Darafsheh, Arash
2018-02-01
Microsphere-assisted imaging can be incorporated onto conventional light microscopes allowing wide-field and flourescence imaging with enhanced resolution. We demonstrated that imaging of specimens containing subdiffraction-limited features is achievable through high-index microspheres embedded in a transparent thin film placed over the specimen. We fabricated novel microsphere-embedded microscope slides composed of barium titanate glass microspheres (with diameter 10-100 μm and refractive index 1.9-2.2) embedded in a transparent polydimethylsiloxane (PDMS) elastomer layer with controllable thickness. We characterized the imaging performance of such microsphere-embedded devices in white-light microscopies, by measuring the imaging resolution, field-of-view, and magnification as a function of microsphere size. Our results inform on the design of novel optical devices, such as microsphere-embedded microscope slides for imaging applications.
Gao, Baojiao; Hu, Hongyan; Guo, Jianfeng; Li, Yanbin
2010-06-01
The crosslinked copolymeric microspheres (HEMA/NVP) of N-vinylpyrrolidone (NVP) and 2-hydroxyethyl methacrylate (HEMA) were prepared using inverse suspension polymerization method. Subsequently, the reaction of methacryloyl chloride with the hydroxyl groups on the surfaces of HEMA/NVP microspheres was performed, leading to the introduction of polymerisable double bonds onto the surfaces of microspheres HEMA/NVP. Afterward, methacrylic acid was allowed to be graft-polymerized on microspheres HEMA/NVP in the manner of "grafting from", resulting in the grafted microspheres PMAA-HEMA/NVP. The grafted microspheres PMAA-HEMA/NVP were fully characterized with several means. The graft-polymerization of MAA on microspheres HEMA/NVP was studied in detail, and the optimal reaction conditions were determined. Thereafter, the adsorption property of the grafted microspheres PMAA-HEMA/NVP for lysozyme as a basic protein model was preliminarily examined to explore the feasibility of removing deleterious basic protein such as density lipoprotein from blood. The experimental results indicate that the PMAA grafting degree on microspheres HEMA/NVP is limited because an enwinding polymer layer as a kinetic barrier on the surfaces of HEMA/NVP microspheres will be formed during the graft-polymerization, and block the graft-polymerization. In order to enhance PMAA grafting degree, reaction temperature, monomer concentration and the used amount of initiator should be effectively controlled. The experimental results also reveal that the grafted microspheres PMAA-HEMA/NVP possess very strong adsorption ability for lysozyme by right of strong electrostatic interaction. Copyright 2010 Elsevier B.V. All rights reserved.
Pluronic F127/chitosan blend microspheres for mucoadhesive drug delivery
NASA Astrophysics Data System (ADS)
Gu, W. Z.; Hu, X. F.
2017-01-01
Pluronic F127/chitosan blend microspheres were prepared via emulsification and cross-linking process using glutaraldehyde as a cross-linker. Compared with chitosan microspheres fabricated under the same experimental conditions, blend microspheres exhibited better physical stability and higher swelling capacity. Puerarin, a traditional Chinese medicine, was incorporated into microparticlesas the model drug. The in vitro release of puerarin from blend microspheres was reduced because of the improved compatibility of the drug with the matrices. According to the results from in vitro adhesion experiments, mucoadhesive behavior of blend microspheres on a mucosa-like surface was similar to that of chitosan microspheres, despite their good ability of anti-protein absorption in solution.
Effect of a freeze-dried CMC/PLGA microsphere matrix of rhBMP-2 on bone healing.
Schrier, J A; Fink, B F; Rodgers, J B; Vasconez, H C; DeLuca, P P
2001-10-07
The hypothesis of this research was that implants of poly(lactide-co-glycolide) (PLGA) microspheres loaded with bone morphogenetic protein-2 (rhBMP-2) and distributed in a freeze-dried carboxymethylcellulose (CMC) matrix would produce more new bone than would matrix implants of non-protein-loaded microspheres or matrix implants of only CMC. To test this hypothesis it was necessary to fashion microsphere-loaded CMC implants that were simple to insert, fit precisely into a defect, and would not elicit swelling. Microspheres were produced via a water-in-oil-in-water double-emulsion system and were loaded with rhBMP-2 by soaking them in a buffered solution of the protein at a concentration of 5.4 mg protein per gram of PLGA. Following recovery of the loaded microspheres by lyophilization, matrices for implantation were prepared by lyophilizing a suspension of the microspheres in 2% CMC in flat-bottom tissue culture plates. Similar matrices were made with 2% CMC and with 2% CMC containing blank microspheres. A full-thickness calvarial defect model in New Zealand white rabbits was used to assess bone growth. Implants fit the defect well, allowing for direct application. Six weeks postsurgery, defects were collected and processed for undecalcified histology. In vitro, 60% of the loaded rhBMP-2 released from devices or microspheres in 5 to 7 days, with the unembedded microspheres releasing faster than those embedded in CMC. In vivo, the rhBMP-2 microspheres greatly enhanced bone healing, whereas nonloaded PLGA microspheres in the CMC implants had little effect. The results showed that a lyophilized device of rhBMP-2/PLGA microspheres in CMC was an effective implantable protein-delivery system for use in bone repair.
Blatchley, E R; Shen, C; Scheible, O K; Robinson, J P; Ragheb, K; Bergstrom, D E; Rokjer, D
2008-02-01
Dyed microspheres have been developed as a new method for validation of ultraviolet (UV) reactor systems. When properly applied, dyed microspheres allow measurement of the UV dose distribution delivered by a photochemical reactor for a given operating condition. Prior to this research, dyed microspheres had only been applied to a bench-scale UV reactor. The goal of this research was to extend the application of dyed microspheres to large-scale reactors. Dyed microsphere tests were conducted on two prototype large-scale UV reactors at the UV Validation and Research Center of New York (UV Center) in Johnstown, NY. All microsphere tests were conducted under conditions that had been used previously in biodosimetry experiments involving two challenge bacteriophage: MS2 and Qbeta. Numerical simulations based on computational fluid dynamics and irradiance field modeling were also performed for the same set of operating conditions used in the microspheres assays. Microsphere tests on the first reactor illustrated difficulties in sample collection and discrimination of microspheres against ambient particles. Changes in sample collection and work-up were implemented in tests conducted on the second reactor that allowed for improvements in microsphere capture and discrimination against the background. Under these conditions, estimates of the UV dose distribution from the microspheres assay were consistent with numerical simulations and the results of biodosimetry, using both challenge organisms. The combined application of dyed microspheres, biodosimetry, and numerical simulation offers the potential to provide a more in-depth description of reactor performance than any of these methods individually, or in combination. This approach also has the potential to substantially reduce uncertainties in reactor validation, thereby leading to better understanding of reactor performance, improvements in reactor design, and decreases in reactor capital and operating costs.
Naproxen-imprinted xerogels in the micro- and nanospherical formsby emulsion technique.
Ornelas, Mariana; Azenha, Manuel; Pereira, Carlos; Silva, A Fernando
2015-11-27
Naproxen-imprinted xerogels in the microspherical and nanospherical forms were prepared by W/O emulsion and microemulsion, respectively. The work evolved from a sol–gel mixture previously reported for bulk synthesis. It was relatively simple to convert the original sol–gel mixture to one amenable to emulsion technique. The microspheres thus produced presented mean diameter of 3.7 μm, surface area ranging 220–340 m2/g, selectivity factor 4.3 (against ibuprofen) and imprinting factor 61. A superior capacity (9.4 μmol/g) was found, when comparing with imprints obtained from similar pre-gelification mixtures. However, slow mass transfer kinetics was deduced from column efficiency results. Concerning the nanospherical format, which constituted the first example of the production of molecularly imprinted xerogels in that format by microemulsion technique, adapting the sol–gel mixture was troublesome. In the end, nanoparticles with diameter in the order of 10 nm were finally obtained, exhibiting good indications of an efficient molecular imprinting process. Future refinements are necessary to solve serious aggregation issues, before moving to more accurate characterization of the binding characteristics or to real applications of the nanospheres.
Pulsed laser triggered high speed microfluidic switch
NASA Astrophysics Data System (ADS)
Wu, Ting-Hsiang; Gao, Lanyu; Chen, Yue; Wei, Kenneth; Chiou, Pei-Yu
2008-10-01
We report a high-speed microfluidic switch capable of achieving a switching time of 10 μs. The switching mechanism is realized by exciting dynamic vapor bubbles with focused laser pulses in a microfluidic polydimethylsiloxane (PDMS) channel. The bubble expansion deforms the elastic PDMS channel wall and squeezes the adjacent sample channel to control its fluid and particle flows as captured by the time-resolved imaging system. A switching of polystyrene microspheres in a Y-shaped channel has also been demonstrated. This ultrafast laser triggered switching mechanism has the potential to advance the sorting speed of state-of-the-art microscale fluorescence activated cell sorting devices.
Conjugate adaptive optics with remote focusing in multiphoton microscopy
NASA Astrophysics Data System (ADS)
Tao, Xiaodong; Lam, Tuwin; Zhu, Bingzhao; Li, Qinggele; Reinig, Marc R.; Kubby, Joel
2018-02-01
The small correction volume for conventional wavefront shaping methods limits their application in biological imaging through scattering media. In this paper, we take advantage of conjugate adaptive optics (CAO) and remote focusing (CAORF) to achieve three-dimensional (3D) scanning through a scattering layer with a single correction. Our results show that the proposed system can provide 10 times wider axial field of view compared with a conventional conjugate AO system when 16,384 segments are used on a spatial light modulator. We demonstrate two-photon imaging with CAORF through mouse skull. The fluorescent microspheres embedded under the scattering layers can be clearly observed after applying the correction.
NASA Technical Reports Server (NTRS)
Blumberg, Seth; Gajraj, Arivalagan; Pennington, Matthew W.; Meiners, Jens-Christian
2005-01-01
Tethered particle microscopy is a powerful tool to study the dynamics of DNA molecules and DNA-protein complexes in single-molecule experiments. We demonstrate that stroboscopic total internal reflection microscopy can be used to characterize the three-dimensional spatiotemporal motion of DNA-tethered particles. By calculating characteristic measures such as symmetry and time constants of the motion, well-formed tethers can be distinguished from defective ones for which the motion is dominated by aberrant surface effects. This improves the reliability of measurements on tether dynamics. For instance, in observations of protein-mediated DNA looping, loop formation is distinguished from adsorption and other nonspecific events.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Silva, Suresh de, E-mail: suresh.desilva@unsw.edu.au; Mackie, Simon; Aslan, Peter
BackgroundIntra-arterial brachytherapy with yttrium-90 ({sup 90}Y) resin microspheres (radioembolization) is a procedure to selectively deliver high-dose radiation to tumors. The purpose of this research was to compare the radioembolic effect of {sup 90}Y-radioembolization versus the embolic effect of bland microspheres in the porcine kidney model.MethodsIn each of six pigs, ~25–33 % of the kidney volume was embolized with {sup 90}Y resin microspheres and an equivalent number of bland microspheres in the contralateral kidney. Kidney volume was estimated visually from contrast-enhanced fluoroscopy imaging. Morphologic and histologic analysis was performed 8–9 weeks after the procedure to assess the locations of the microspheres and extentmore » of tissue necrosis from {sup 90}Y-radioembolization and bland embolization. A semi-quantified evaluation of the non-acute peri-particle and perivascular tissue reaction was conducted. All guidelines for the care and use of animals were followed.ResultsKidneys embolized with {sup 90}Y-radioembolization decreased in mass by 30–70 % versus the contralateral kidney embolized with bland microspheres. These kidneys showed significant necrosis/fibrosis, avascularization, and glomerular atrophy in the immediate vicinity of the {sup 90}Y resin microspheres. By contrast, glomerular changes were not observed, even with clusters of bland microspheres in afferent arterioles. Evidence of a foreign body reaction was recorded in some kidneys with bland microspheres, and subcapsular scarring/infarction only with the highest load (4.96 × 10{sup 6}) of bland microspheres.ConclusionThis study showed that radioembolization with {sup 90}Y resin microspheres produces localized necrosis/fibrosis and loss of kidney mass in a porcine kidney model. This result supports the study of {sup 90}Y resin microspheres for the localized treatment of kidney tumors.« less
Formulation and evaluation of Bacillus coagulans-loaded hypromellose mucoadhesive microspheres.
Alli, Sk Md Athar
2011-01-01
Development of a novel delivery system has been attempted to deliver viable probiotic cells into the gut for a prolonged period of time while maintaining high numbers of viable cells within the formulation throughout the shelf-life of the product and during the gastrointestinal transit. Core mucoadhesive microspheres of Bacillus coagulans were developed employing several grades of hypromellose, a mucoadhesive polymer, following coacervation and phase separation technique and were subsequently enteric-coated with hypromellose phthalate. Microspheres were evaluated for percent yield; entrapment efficiency; in vitro swelling; surface morphology; particle size, size distribution, and zeta potential; flow property, mucoadhesion property by the ex vivo mucoadhesive strength test and the in vitro wash off test; in vitro release profile and release kinetic; in vivo probiotic activity; and stability. The values for the kinetic constant and regression coefficient of model-dependent approaches and the difference factor (f(1)), the similarity factor (f(2)), and the Rescigno index (ξ(1) and ξ(2)) of model independent approaches were determined for comparing in vitro dissolution profiles. Freeze dried B. coagulans cells were successfully formulated as enteric-coated mucoadhesive microspheres with satisfactory physical structure and yield. The viability of B. coagulans was maintained in the simulated gastric conditions and during processing; in simulated intestinal conditions exhibiting mucoadhesion, and controlling and extending the viable cell release following zero-order; and was satisfactorily stable at room temperature. Test results depict statistically significant effects of the hypromellose grade and their concentration on the performance and release profile of formulations.
Sustained release effects of berberine-loaded chitosan microspheres on in vitro chondrocyte culture.
Zhou, Yan; Liu, Shiqing; Ming, Jianghua; Li, Yaming; Deng, Ming; He, Bin
2017-10-01
The low bioavailability and short biological half-life of berberine chloride (BBR) negatively affect the protective role of this compound against osteoarthritis (OA). The present study was performed to evaluate the effectiveness of sustained BBR release system. Novel BBR-loaded chitosan microspheres (BBR-loaded CMs) were successfully synthesized using an ionic cross-linking method for sustained release. The basic characteristics of the prepared microspheres were subsequently evaluated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) techniques, encapsulation efficiency (EE), and in vitro release experiments. BBR-loaded CMs displayed spherical forms to encapsulate a considerable quantity of BBR (100.8 ± 2.7 mg/g); these microspheres also exhibited an ideal releasing profile. The FT-IR spectra and XRD results revealed that BBR-loaded CMs were successfully synthesized via electrostatic interaction. In vitro experiments further showed that BBR-loaded CMs significantly inhibited sodium nitroprusside (SNP)-stimulated chondrocyte apoptosis as well as cytoskeletal remodeling, and led to increasing mitochondrial membrane potential and maintaining the nuclear morphology. BBR-loaded CMs exerted markedly higher anti-apoptotic activity in the treatment of OA, and markedly inhibited the protein expression levels of caspase-3, a disintegrin, and metalloproteinase with thrombospondin motifs (ADAMTS)-5 and matrix metalloproteinase (MMP)-13 induced by SNP in rat articular chondrocytes, compared with free BBR at equivalent concentration. Therefore, novel BBR-loaded CMs may offer potential for application in the treatment of OA.
Formulation and evaluation of Bacillus coagulans-loaded hypromellose mucoadhesive microspheres
Alli, Sk Md Athar
2011-01-01
Development of a novel delivery system has been attempted to deliver viable probiotic cells into the gut for a prolonged period of time while maintaining high numbers of viable cells within the formulation throughout the shelf-life of the product and during the gastrointestinal transit. Core mucoadhesive microspheres of Bacillus coagulans were developed employing several grades of hypromellose, a mucoadhesive polymer, following coacervation and phase separation technique and were subsequently enteric-coated with hypromellose phthalate. Microspheres were evaluated for percent yield; entrapment efficiency; in vitro swelling; surface morphology; particle size, size distribution, and zeta potential; flow property, mucoadhesion property by the ex vivo mucoadhesive strength test and the in vitro wash off test; in vitro release profile and release kinetic; in vivo probiotic activity; and stability. The values for the kinetic constant and regression coefficient of model-dependent approaches and the difference factor (f1), the similarity factor (f2), and the Rescigno index (ξ1 and ξ2) of model independent approaches were determined for comparing in vitro dissolution profiles. Freeze dried B. coagulans cells were successfully formulated as enteric-coated mucoadhesive microspheres with satisfactory physical structure and yield. The viability of B. coagulans was maintained in the simulated gastric conditions and during processing; in simulated intestinal conditions exhibiting mucoadhesion, and controlling and extending the viable cell release following zero-order; and was satisfactorily stable at room temperature. Test results depict statistically significant effects of the hypromellose grade and their concentration on the performance and release profile of formulations. PMID:21674019
Sasiak, A B; Bolgiano, B; Crane, D T; Hockley, D J; Corbel, M J; Sesardic, D
2000-11-22
The purpose of this study was to investigate the utility of various in vitro and in vivo methods to assess the stability of experimental vaccines containing tetanus toxoid (TT) within PLGA microspheres. In vitro, the breakdown of the encapsulating polymers into their acid components led to changes in the structure of TT, as determined by the physico-chemical methods, rendering it undetectable by capture ELISA and altering its structural integrity. The changes in TT were directly related to increasing acidity of the vaccine supernate. Purified toxoid (not encapsulated) exposed to low pH (2.5) underwent similar changes but re-neutralisation of buffer containing free toxoid, even after one week at pH 2.5 led to some re-folding of protein as determined by fluorescence spectroscopy and gel filtration chromatography. The microencapsulated vaccines were still able to generate an antibody response in mice even after prolonged pre-incubation at 37 degrees C and the apparent absence of detectable toxoid in the vaccine supernate. Electron microscopy demonstrated differences in the amount of degradation between different formulations of microspheres. Vaccines that had retained their spherical morphology after incubation in vitro for up to 28 days were able to induce protective antibodies response equal to that of freshly prepared vaccines, which indicates that the toxoid within intact microspheres remained immunogenic. Immunochemical and physico-chemical detection methods, performed on antigen released from PLGA vaccines in vitro, are valuable in providing information on product characteristics but may not be able to predict effectiveness and should be used with in vivo methods to evaluate the stability of such formulations.
Liu, Hongmei; Lu, Anxiang; Fu, Hailong; Li, Bingru; Yang, Meihua; Wang, Jihua; Luan, Yunxia
2018-06-12
A novel adsorbent is described for magnetic solid-phase extraction (MSPE) of the aflatoxins AFB 1 and AFB 2 (AFBs). Magnetic agarose microspheres (MAMs) were functionalized with an aptamer to bind the AFBs which then were quantified by HPLC and on-line post-column photochemical derivatization with fluorescence detection. Streptavidin-conjugated MAMs were synthesized first by a highly reproducible strategy. They possess strong magnetism and high surface area. The MAMs were characterized by transmission electron microscopy, scanning electron microscopy, optical microscopy, laser diffraction particle size analyzer, Fourier transform infrared spectrometry, vibrating sample magnetometry and laser scanning confocal microscopy. Then, the AFB-aptamers were immobilized on MAMs through biotin-streptavidin interaction. Finally, the MSPE is performed by suspending the aptamer-modified MAMs in the sample. They are then collected by an external magnetic field and the AFBs are eluted with methanol/buffer (20:80). Several parameters affecting the coupling, capturing and eluting efficiency were optimized. Under the optimized conditions, the method is fast, has good linearity, high selectivity, and sensitivity. The LODs are 25 pg·mL -1 for AFB 1 and 10 pg·mL -1 for AFB 2 . The binding capacity is 350 ± 8 ng·g -1 for AFB 1 and 384 ± 8 ng·g -1 for AFB 2 , and the precision of the assay is <8%. The method was successfully applied to the analysis of AFBs in spiked maize samples. Graphical abstract Schematic of novel aptamer functionalized magnetic agarose microspheres (Apt-MAM) as magnetic adsorbents for simultaneous and specific affinity capture of aflatoxins B 1 and B 2 (AFBs).
Koyama, Hidenori; Okada, Masahiro; Tanaka, Shinji; Shoji, Tetsuo; Emoto, Masanori; Furuzono, Tsutomu; Nishizawa, Yoshiki; Inaba, Masaaki
2012-01-01
Background Clinical trials demonstrate the effectiveness of cell-based therapeutic angiogenesis in patients with severe ischemic diseases; however, their success remains limited. Maintaining transplanted cells in place are expected to augment the cell-based therapeutic angiogenesis. We have reported that nano-hydroxyapatite (HAp) coating on medical devices shows marked cell adhesiveness. Using this nanotechnology, HAp-coated poly(l-lactic acid) (PLLA) microspheres, named nano-scaffold (NS), were generated as a non-biological, biodegradable and injectable cell scaffold. We investigate the effectiveness of NS on cell-based therapeutic angiogenesis. Methods and Results Bone marrow mononuclear cells (BMNC) and NS or control PLLA microspheres (LA) were intramuscularly co-implanted into mice ischemic hindlimbs. When BMNC derived from enhanced green fluorescent protein (EGFP)-transgenic mice were injected into ischemic muscle, the muscle GFP level in NS+BMNC group was approximate fivefold higher than that in BMNC or LA+BMNC groups seven days after operation. Kaplan-Meier analysis demonstrated that NS+BMNC markedly prevented hindlimb necrosis (P<0.05 vs. BMNC or LA+BMNC). NS+BMNC revealed much higher induction of angiogenesis in ischemic tissues and collateral blood flow confirmed by three-dimensional computed tomography angiography than those of BMNC or LA+BMNC groups. NS-enhanced therapeutic angiogenesis and arteriogenesis showed good correlations with increased intramuscular levels of vascular endothelial growth factor and fibroblast growth factor-2. NS co-implantation also prevented apoptotic cell death of transplanted cells, resulting in prolonged cell retention. Conclusion A novel and feasible injectable cell scaffold potentiates cell-based therapeutic angiogenesis, which could be extremely useful for the treatment of severe ischemic disorders. PMID:22529991
Direct Compositional Characterization of (U,Th)O2 Powders, Microspheres, and Pellets Using TXRF.
Dhara, Sangita; Prabhat, Parimal; Misra, N L
2015-10-20
A total reflection X-ray fluorescence (TXRF) analysis method for direct compositional characterization of sintered and green (U,Th)O2 samples in different forms (e.g., pellets, powders, and microspheres) without sample dissolution has been developed for the first time. The methodology involves transfer of only a few nanograms of the sample on the TXRF sample support by gently rubbing the samples on supports or taking their tiny uniform slurry in collodion on the sample support, drying them to make thin film, and measuring the TXRF spectra of the specimens thus prepared. This approach minimizes the matrix effects. Uranium determinations from the TXRF spectra of such specimens were made with respect to thorium, considering it as an internal standard. Samples having uranium atom percent (at%) from 0 to 100 in (U,Th)O2 were analyzed for uranium in comparison to thorium. The results showed an average precision of 2.6% (RSD, 2σ, n = 8). The TXRF-determined results deviated from expected values within 5%. The TXRF results were compared with those of biamperometry with good agreement. The lattice parameters of the solid solutions were calculated using their XRD patterns. A good correlation between lattice parameters and TXRF-determined U at% and between TXRF-determined U at% and expected U at%, calculated on the basis of preparation of (U,Th)O2 solid solutions, was obtained. The developed method is capable of analyzing (U,Th)O2 samples directly with almost negligible sample preparation and is well suited for radioactive samples. The present study suggests that this method can be extended for the determination of U,Th and Pu in other nuclear fuel materials (e.g., nitrides, carbides, etc.) in the form of pellets, powders, and microspheres after suitable modifications in sample handling procedure.
Zhang, Huan; Wang, Weimei; Li, Haoran; Peng, Yi; Zhang, Zhiqing
2018-01-01
Insulin-loaded microspheres were prepared by alternating deposition film layers that were composed of insulin and poly(vinyl sulfate) potassium on the surface of poly(lactic acid) (PLA) microspheres. The preparation of the insulin-loaded microspheres was optimized by an orthogonal test design, and the relationship between drug loading (DL) and film layers was studied. The particle size, DL and encapsulation efficiency of the obtained insulin-loaded microspheres with 10 films were 5.25 ± 0.15 µm, 111.33 ± 1.15 mg/g and 33.7 ± 0.19%, respectively. Following this, the physical characteristics of the insulin-loaded microspheres were investigated. The results from scanning electron microscopy and a laser particle size analyzer (LPSA) indicated the spherical morphology, rough surface and increasing particle sizes of the insulin-loaded microspheres, which were compared to those of PLA microspheres. An in vitro release study showed that the insulin-loaded microspheres were stable in HCl solution (pH 1.0) and released insulin slowly in phosphate-buffered solution (pH 6.8). Finally, the drug efficacy of the prepared insulin-loaded microspheres via oral administration was evaluated in rats with diabetes induced by streptozotocin, and an obvious dose-dependent hypoglycemic effect was observed. This preliminary data could illustrate the prospect of using microspheres for the oral delivery of insulin.
Chen, Xingtao; Lv, Guoyu; Zhang, Jue; Tang, Songchao; Yan, Yonggang; Wu, Zhaoying; Su, Jiacan; Wei, Jie
2014-01-01
A multi-(amino acid) copolymer (MAC) based on ω-aminocaproic acid, γ-aminobutyric acid, L-alanine, L-lysine, L-glutamate, and hydroxyproline was synthetized, and MAC microspheres encapsulating bovine serum albumin (BSA) were prepared by a double-emulsion solvent extraction method. The experimental results show that various preparation parameters including surfactant ratio of Tween 80 to Span 80, surfactant concentration, benzyl alcohol in the external water phase, and polymer concentration had obvious effects on the particle size, morphology, and encapsulation efficiency of the BSA-loaded microspheres. The sizes of BSA-loaded microspheres ranged from 60.2 μm to 79.7 μm, showing different degrees of porous structure. The encapsulation efficiency of BSA-loaded microspheres also ranged from 38.8% to 50.8%. BSA release from microspheres showed the classic biphasic profile, which was governed by diffusion and polymer erosion. The initial burst release of BSA from microspheres at the first week followed by constant slow release for the next 7 weeks were observed. BSA-loaded microspheres could degrade gradually in phosphate buffered saline buffer with pH value maintained at around 7.1 during 8 weeks incubation, suggesting that microsphere degradation did not cause a dramatic pH drop in phosphate buffered saline buffer because no acidic degradation products were released from the microspheres. Therefore, the MAC microspheres might have great potential as carriers for protein delivery. PMID:24855351
Synthesis and catalytic performance of SiO2@Ni and hollow Ni microspheres
NASA Astrophysics Data System (ADS)
Liu, Xin; Liu, Yanhua; Shi, Xueting; Yu, Zhengyang; Feng, Libang
2016-11-01
Nickel (Ni) catalyst has been widely used in catalytic reducing reactions such as catalytic hydrogenation of organic compounds and catalytic reduction of organic dyes. However, the catalytic efficiency of pure Ni is low. In order to improve the catalytic performance, Ni nanoparticle-loaded microspheres can be developed. In this study, we have prepared Ni nanoparticle-loaded microspheres (SiO2@Ni) and hollow Ni microspheres using two-step method. SiO2@Ni microspheres with raspberry-like morphology and core-shell structure are synthesized successfully using SiO2 microsphere as a template and Ni2+ ions are adsorbed onto SiO2 surfaces via electrostatic interaction and then reduced and deposited on surfaces of SiO2 microspheres. Next, the SiO2 cores are removed by NaOH etching and the hollow Ni microspheres are prepared. The NaOH etching time does no have much influence on the crystal structure, shape, and surface morphology of SiO2@Ni; however, it can change the phase composition evidently. The hollow Ni microspheres are obtained when the NaOH etching time reaches 10 h and above. The as-synthesized SiO2@Ni microspheres exhibit much higher catalytic performance than the hollow Ni microspheres and pure Ni nanoparticles in the catalytic reduction of methylene blue. Meanwhile, the SiO2@Ni catalyst has high stability and hence it can be recycled for reuse.
Size effect of optical silica microsphere pressure sensors
NASA Astrophysics Data System (ADS)
Jiao, Xinbing; Hao, Ruirui; Pan, Qian; Zhao, Xinwei; Bai, Xue
2018-07-01
Two types of optical pressure sensors with silica microspheres are proposed. The size effect of optical silica microsphere pressure sensors is studied by using a single-wavelength laser beam and polarimeters. The silica microspheres with diameters of 1.0 μm, 1.5 μm and 2.0 μm are prepared on garnet substrates by a self-assembly method. The pressure and the optical properties of the silica microspheres are measured by a resistance strain sensor and Thorlabs Stokes polarimeters as a function of the external direct current (DC) voltage. The optical silica microsphere sensor in transmission mode is suitable for pressure measuring. The results show that the pressure increases, while the diameter of the silica microspheres decreases. The maximum internal pressure can reach up to 7.3 × 107 Pa when the diameter of the silica microspheres is 1.0 μm.
Xu, Leimeng; Chen, Jiawei; Song, Jizhong; Li, Jianhai; Xue, Jie; Dong, Yuhui; Cai, Bo; Shan, Qingsong; Han, Boning; Zeng, Haibo
2017-08-09
Novel fluorescence with highly covert and reliable features is quite desirable to combat the sophisticated counterfeiters. Herein, we report a simultaneously triple-modal fluorescent characteristic of CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 by the excitation of thermal, ultraviolet (UV) and infrared (IR) light for the first time, which can be applied for the multiple modal anti-counterfeiting codes. The diphasic structure CsPbBr 3 @Cs 4 PbBr 6 nanocrystals (NCs) was synthesized via the typical reprecipitation method followed by uniformly encapsulation into silica microspheres. Cubic CsPbBr 3 is responsible for the functions of anti-counterfeiting, while Cs 4 PbBr 6 crystalline and SiO 2 are mainly to protect unstable CsPbBr 3 NCs from being destroyed by ambient conditions. The as-prepared CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 NCs possess improved stability and are capable of forming printable ink with organic binders for patterns. Interestingly, the fluorescence of diphasic CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 capsule patterns can be reversibly switched by the heating, UV, and IR light irradiation, which has been applied as triple-modal fluorescent anti-counterfeiting codes. The results demonstrate that the perovskite@silica capsules are highly promising for myriad applications in areas such as fluorescent anti-counterfeiting, optoelectronic devices, medical diagnosis, and biological imaging.
Wang, Haiying; Wang, Hong; Chen, Shaopei; Dzakah, Emmanuel E; Kang, Keren; Wang, Jihua; Wang, Jufang
2015-04-15
Procalcitonin (PCT) has been recognized as a biomarker in severe inflammation, infection and sepsis. PCT detection in serum requires sensitive and specific antibodies. In this study, we generated monoclonal antibodies (mAbs) and developed fluorescent immunochromatographic assay for PCT detection. Human recombinant PCT was used as immunogen. mAbs against PCT were developed and applied to fluorescent immunochromatographic assay for PCT detection in clinical samples. Out of 35 hybridoma cell lines secreting antibodies against the recombinant PCT, five sensitive and specific cell lines were selected and designated as F6, G2, C2, D2 and E5. All these antibodies have no cross reaction with calcitonin or calcitonin gene-related peptides (CGRP). After screening for pairing, mAb F6 was labeled with fluorescent microspheres and C2 was coated on a nitrocellulose membrane for immunochromatographic test. All 35 clinical samples were detected by the mAb F6-C2 test strips and the bioMérieux PCT assay. The test strips showed high specificity and sensitivity for PCT. Good correlation was observed between our immunochromatographic test strips and the bioMérieux PCT assay (R(2):0.986). These newly developed anti-PCT mAbs and fluorescent immunochromatographic assay can serve as important diagnostic tools for a fast, reliable and point-of-care testing for easy determination of PCT in serum and diagnosis of bacterial infection, inflammation or sepsis. Copyright © 2015 Elsevier B.V. All rights reserved.
The 400 microsphere per piece "rule" does not apply to all blood flow studies.
Polissar, N L; Stanford, D C; Glenny, R W
2000-01-01
Microsphere experiments are useful in measuring regional organ perfusion as well as heterogeneity of blood flow within organs and correlation of perfusion between organ pieces at different time points. A 400 microspheres/piece "rule" is often used in planning experiments or to determine whether experiments are valid. This rule is based on the statement that 400 microspheres must lodge in a region for 95% confidence that the observed flow in the region is within 10% of the true flow. The 400 microspheres precision rule, however, only applies to measurements of perfusion to a single region or organ piece. Examples, simulations, and an animal experiment were carried out to show that good precision for measurements of heterogeneity and correlation can be obtained from many experiments with <400 microspheres/piece. Furthermore, methods were developed and tested for correcting the observed heterogeneity and correlation to remove the Poisson "noise" due to discrete microsphere measurements. The animal experiment shows adjusted values of heterogeneity and correlation that are in close agreement for measurements made with many or few microspheres/piece. Simulations demonstrate that the adjusted values are accurate for a variety of experiments with far fewer than 400 microspheres/piece. Thus the 400 microspheres rule does not apply to many experiments. A "rule of thumb" is that experiments with a total of at least 15,000 microspheres, for all pieces combined, are very likely to yield accurate estimates of heterogeneity. Experiments with a total of at least 25,000 microspheres are very likely to yield accurate estimates of correlation coefficients.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zhao, Xinyu; School of Petrochemical Engineering, Shenyang University of Technology, Liaoyang 111003; Cao, Minhua, E-mail: caomh@bit.edu.cn
2013-06-01
Graphical abstract: MoO{sub 3} hollow microspheres were synthesized via a facile and template-free solvothermal route and subsequent heat treatment in air. The MoO{sub 3} hollow microspheres exhibit an improved lithium storage and gas-sensing performance. Highlights: ► Hollow MoO{sub 3} microspheres were synthesized by thermal oxidation of hollow MoO{sub 2}. ► The MoO{sub 3} hollow microspheres have a relatively high specific surface area. ► The MoO{sub 3} hollow microspheres exhibit improved lithium storage performance. ► The MoO{sub 3} hollow microspheres show good responses to ammonia gas. - Abstract: In this paper, MoO{sub 3} hollow microspheres were synthesized via a facile andmore » template-free solvothermal route and subsequent heat treatment in air. The MoO{sub 3} hollow microspheres have a relatively high specific surface area, and with such a feature, the as-synthesized MoO{sub 3} hollow microspheres have potential applications in Li-ion battery and gas-sensor. When tested as a Li-storage anode material, the MoO{sub 3} hollow microspheres show a higher discharge capacity of 1377.1 mA h g{sup −1} in the first discharge and a high reversible capacity of 780 mA h g{sup −1} after 100 cycles at a rate of 1 C. Furthermore, as a gas sensing material, the MoO{sub 3} hollow microspheres exhibit an improved sensitivity and short response/recovery time to trace levels of ammonia gas.« less
NASA Astrophysics Data System (ADS)
Yu, Bing; Cong, Hailin; Liu, Xuesong; Ren, Yumin; Wang, Jilei; Zhang, Lixin; Tang, Jianguo; Ma, Yurong; Akasaka, Takeshi
2013-09-01
An effective microfluidic method to fabricate monodisperse polyethylene glycol (PEG) hydrogel composite microspheres with tunable dimensions and properties is reported in this paper. A T-junction microfluidic chip equipped with rounded channels and online photopolymerization system is applied for the microsphere microfabrication. The shape and size of the microspheres are well controlled by the rounded channels and PEG prepolymer/silicon oil flow rate ratios. The obtained PEG/aspirin composite microspheres exhibit a sustained release of aspirin for a wide time range; the obtained PEG/Fe3O4 nanocomposite microspheres exhibit excellent magnetic properties; and the obtained binary PEG/dye composite microspheres show the ability to synchronously load two functional components in the same peanut-shaped or Janus hydrogel particles.
Fabrication of glass microspheres with conducting surfaces
Elsholz, William E.
1984-01-01
A method for making hollow glass microspheres with conducting surfaces by adding a conducting vapor to a region of the glass fabrication furnace. As droplets or particles of glass forming material pass through multiple zones of different temperature in a glass fabrication furnace, and are transformed into hollow glass microspheres, the microspheres pass through a region of conducting vapor, forming a conducting coating on the surface of the microspheres.
Fabrication of glass microspheres with conducting surfaces
Elsholz, W.E.
1982-09-30
A method for making hollow glass microspheres with conducting surfaces by adding a conducting vapor to a region of the glass fabrication furnace. As droplets or particles of glass forming material pass through multiple zones of different temperature in a glass fabrication furnace, and are transformed into hollow glass microspheres, the microspheres pass through a region of conducting vapor, forming a conducting coating on the surface of the microspheres.
Polarization Dependent Whispering Gallery Modes in Microspheres
NASA Technical Reports Server (NTRS)
Adamovsky, Grigory (Inventor); Wrbanek, Susan Y. (Inventor)
2016-01-01
A tunable resonant system is provided and includes a microsphere that receives an incident portion of a light beam generated via a light source, the light beam having a fundamental mode, a waveguide medium that transmits the light beam from the light source to the microsphere, and a polarizer disposed in a path of the waveguide between the light source and the microsphere. The incident portion of the light beam creates a fundamental resonance inside the microsphere. A change in a normalized frequency of the wavelength creates a secondary mode in the waveguide and the secondary mode creates a secondary resonance inside the microsphere.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Chen, Limiao, E-mail: chenlimiao@csu.edu.cn; College of Chemistry and Chemical Engineering, Central South University, Changsha 410083; Wu, Min
The semiconductor nanostructures decorated with noble metals have attracted increasing attention due to their interesting physical and chemical properties. In this work, urchin-like monoclinic (m-) LaVO{sub 4} microspheres were prepared by a hydrothermal method and used as a template to fabricate Ag nanoparticle-decorated m-LaVO{sub 4} composites. The morphology and structure were characterized by transmission electron microscope, high-resolution transmission electron microscope, scanning electron microscope, and energy-dispersive X-ray. It was found that Ag nanoparticles with narrow size distribution were uniformly loaded on urchin-like m-LaVO{sub 4} microspheres, and the resulted composite microspheres showed distinct surface plasmon absorption band compared to pure m-LaVO{sub 4}more » microspheres. Photocatalytic activities of as-prepared samples were examined by studying the degradation of methyl orange solutions under visible-light irradiation (> 400 nm). Results clearly showed that urchin-like m-LaVO{sub 4}/Ag microspheres possess much higher photocatalytic activity than pure m-LaVO{sub 4} microspheres and P25. - Highlights: • m-LaVO{sub 4}/Ag composites microspheres were fabricated by a hydrothermal method. • m-LaVO{sub 4} microspheres show higher photocatalytic activity than m-LaVO{sub 4} microspheres. • m-LaVO{sub 4}/Ag microspheres exhibit a good stability.« less
Bioactive and biodegradable silica biomaterial for bone regeneration.
Wang, Shunfeng; Wang, Xiaohong; Draenert, Florian G; Albert, Olga; Schröder, Heinz C; Mailänder, Volker; Mitov, Gergo; Müller, Werner E G
2014-10-01
Biosilica, a biocompatible, natural inorganic polymer that is formed by an enzymatic, silicatein-mediated reaction in siliceous sponges to build up their inorganic skeleton, has been shown to be morphogenetically active and to induce mineralization of human osteoblast-like cells (SaOS-2) in vitro. In the present study, we prepared beads (microspheres) by encapsulation of β-tricalcium phosphate [β-TCP], either alone (control) or supplemented with silica or silicatein, into the biodegradable copolymer poly(d,l-lactide-co-glycolide) [PLGA]. Under the conditions used, ≈5% β-TCP, ≈9% silica, and 0.32μg/mg of silicatein were entrapped into the PLGA microspheres (diameter≈800μm). Determination of the biocompatibility of the β-TCP microspheres, supplemented with silica or silicatein, revealed no toxicity in the MTT based cell viability assay using SaOS-2 cells. The adherence of SaOS-2 cells to the surface of silica-containing microspheres was higher than for microspheres, containing only β-TCP. In addition, the silica-containing β-TCP microspheres and even more pronounced, a 1:1 mixture of microspheres containing β-TCP and silica, and β-TCP and silicatein, were found to strongly enhance the mineral deposition by SaOS-2 cells. Using these microspheres, first animal experiments with silica/biosilica were performed in female, adult New Zealand White rabbits to study the effect of the inorganic polymer on bone regeneration in vivo. The microspheres were implanted into 5mm thick holes, drilled into the femur of the animals, applying a bilateral comparison study design (3 test groups with 4-8 animals each). The control implant on one of the two hind legs contained microspheres with only β-TCP, while the test implant on the corresponding leg consisted either of microspheres containing β-TCP and silica, or a 1:1 mixture of microspheres, supplemented with β-TCP and silica, and β-TCP and silicatein. The results revealed that tissue/bone sections of silica containing implants and implants, composed of a 1:1 mixture of silica-containing microspheres and silicatein-containing microspheres, show an enhanced regeneration of bone tissue around the microspheres, compared to the control implants containing only β-TCP. The formation of new bone induced by the microspheres is also evident from measurements of the stiffness/reduced Young's modulus of the regenerated bone tissue. The reduced Young's modulus of the regenerating bone tissue around the implants was markedly higher for the silica-containing microspheres (1.1MPa), and even more for the 1:1 mixture of the silica- and silicatein-containing microspheres (1.4MPa), compared to the β-TCP microsphere controls (0.4MPa). We propose that based on their morphogenetic activity on bone-forming cells in vitro and the results of the animal experiments presented here, silica/biosilica-based scaffolds are promising materials for bone repair/regeneration. Copyright © 2014 Elsevier Inc. All rights reserved.
Jamre, Mina; Shamsaei, Mojtaba; Erfani, Mostafa; Sadjadi, Sodeh; Ghannadi Maragheh, Mohammad
2018-04-12
Radioembolization with radioactive microspheres has been an effective method for the treatment of liver lesions. The aim of this study was to prepare carrier-free 188 Re loaded poly (L-lactic acid) (PLLA) microspheres through 188 Re sulfide colloidal nanoparticles ( 188 Re-SC nanoparticles). The formation of 188 Re-SC nanoparticles was confirmed by ultraviolet-visible spectrophotometry. The labeling yield of 188 Re-SC nanoparticles was verified using the RTLC method. Effects of synthesis parameters on morphology and size of prepared 188 Re-sulfide colloidal-PLLA microspheres ( 188 Re-SC-PLLA microspheres) were studied by scanning electron microscopy. In vitro stability of 188 Re-SC-PLLA microspheres was investigated in normal saline at room temperature and in human serum at 37°C. In vivo distribution studies and gamma camera imaging were performed in healthy BALB/c mice. The microspheres could be prepared with sizes between 13 and 48 μm (modal value 29 μm) and radiolabeling efficiency >99%. After incubation, the microspheres were found stable in vitro up to 72 hours. The biodistribution after intravenous injection in healthy BALB/c mice showed high accumulation in lung as a first capture pathway organ for microsphere followed by great retention over 48 hours for these microspheres. These data show that 188 Re-SC-PLLA microspheres are suitable candidate for clinical studies. Copyright © 2018 John Wiley & Sons, Ltd.
Microspheres and their methods of preparation
Bose, Anima B; Yang, Junbing
2015-03-24
Carbon microspheres are doped with boron to enhance the electrical and physical properties of the microspheres. The boron-doped carbon microspheres are formed by a CVD process in which a catalyst, carbon source and boron source are evaporated, heated and deposited onto an inert substrate.
Mesoporous metal oxide microsphere electrode compositions and their methods of making
Paranthaman, Mariappan Parans; Liu, Hansan; Brown, Gilbert M.; Sun, Xiao-Guang; Bi, Zhonghe
2016-12-06
Compositions and methods of making are provided for mesoporous metal oxide microspheres electrodes. The mesoporous metal oxide microsphere compositions comprise (a) microspheres with an average diameter between 200 nanometers (nm) and 10 micrometers (.mu.m); (b) mesopores on the surface and interior of the microspheres, wherein the mesopores have an average diameter between 1 nm and 50 nm and the microspheres have a surface area between 50 m.sup.2/g and 500 m.sup.2/g. The methods of making comprise forming composite powders. The methods may also comprise refluxing the composite powders in a basic solution to form an etched powder, washing the etched powder with an acid to form a hydrated metal oxide, and heat-treating the hydrated metal oxide to form mesoporous metal oxide microspheres.
Polymer-coated albumin microspheres as carriers for intravascular tumour targeting of cisplatin.
Verrijk, R; Smolders, I J; McVie, J G; Begg, A C
1991-01-01
We used a poly-lactide-co-glycolide polymer (PLAGA 50:50) to formulate cisplatin (cDDP) into microspheres designed for intravascular administration. Two systems were developed. PLAGA-coated albumin microspheres and microspheres consisting of PLAGA only. PLAGA-coated microspheres displayed a mean diameter of 31.8 +/- 0.9 microns and a payload of 7.5% cDDP (w/w). Solid PLAGA microspheres exhibited a mean diameter of 19.4 +/- 0.6 microns and a payload of 20% cDDP. Release characteristics and in vitro effects on L1210 leukemia and B16 melanoma cell lines were investigated. Both types of microsphere overcame the initial rapid release of cDDP (burst effect), and PLAGA-coated albumin microspheres also showed a lag phase of approximately 30 min before cDDP release began. PLAGA-coated albumin microspheres released most of their payload through diffusion, and the coating eventually cracked after 7 days' incubation in saline supplemented with 0.1% Tween at 37 degrees C, enabling the release of any cDDP remaining. Effects of platinum, pre-released from PLAGA-coated albumin microspheres on the in vitro growth of L1210 cells were comparable with those of standard formulations (dissolved) of cDDP. Material released from non-drug-loaded PLAGA microspheres had no effect on L1210 cell growth, suggesting the absence of cytotoxic compounds in the matrix. The colony-forming ability of B16 cells was also equally inhibited by standard cDDP and pre-released drug. These studies show that formulation of cDDP in PLAGA-based microspheres prevents the rapid burst effect of cDDP seen in previous preparations and offers an improved system of administration for hepatic artery infusion or adjuvant therapy, enabling better clinical handling and the promise of a higher ratio of tumour tissue to normal tissue.
Principles and applications of flow cytometry and cell sorting in companion animal medicine.
Wilkerson, Melinda J
2012-01-01
Flow cytometry measures multiple characteristic of single cells using light scatter properties and fluorescence properties of fluorescent probes with specificity to cellular constituents. The use of flow cytometry in the veterinary clinical laboratory has become more routine in veterinary diagnostic laboratories and institutions (http://www.vet.k-state.edu/depts/dmp/service/immunology/index.htm), and reference laboratories. The most common applications in small animal medicine includes quantitation of erythrocytes and leukocytes in automated hematology instruments, detection of antibodies to erythrocytes and platelets in cases of immune-mediated diseases, immunophenotyping of leukocytes and lymphocytes in immunodeficiency syndromes, or leukemias and lymphomas. DNA content analysis to identify aneuploidy or replicating cells in tumor preparations has not gained routine acceptance because of the variability of prognostic results. Other applications including cell sorting and multiplexing using microspheres are potential assays of the future once they become validated and the instrumentation footprint becomes more and more compact, less expensive, and easier to use.
Laser induced fluorescence technique for detecting organic matter in East China Sea
NASA Astrophysics Data System (ADS)
Chen, Peng; Wang, Tianyu; Pan, Delu; Huang, Haiqing
2017-10-01
A laser induced fluorescence (LIF) technique for fast diagnosing chromophoric dissolved organic matter (CDOM) in water is discussed. We have developed a new field-portable laser fluorometer for rapid fluorescence measurements. In addtion, the fluorescence spectral characteristics of fluorescent constituents (e.g., CDOM, chlorophyll-a) were analyzed with a spectral deconvolution method of bi-Gaussian peak function. In situ measurements by the LIF technique compared well with values measured by conventional spectrophotometer method in laboratory. A significant correlation (R2 = 0.93) was observed between fluorescence by the technique and absorption by laboratory spectrophotometer. Influence of temperature variation on LIF measurement was investigated in lab and a temperature coefficient was deduced for fluorescence correction. Distributions of CDOM fluorescence measured using this technique in the East China Sea coast were presented. The in situ result demonstrated the utility of the LIF technique for rapid detecting dissolved organic matter.
Xia, Hongjun; Wan, Guangping; Zhao, Junlong; Liu, Jiawei; Bai, Quan
2016-11-04
High performance liquid chromatography (HPLC) is a kind of efficient separation technology and has been used widely in many fields. Micro-sized porous silica microspheres as the most popular matrix have been used for fast separation and analysis in HPLC. In this paper, the monodisperse large-porous silica microspheres with controllable size and structure were successfully synthesized with polymer microspheres as the templates and characterized. First, the poly(glycidyl methacrylate-co-ethyleneglycol dimethacrylate) microspheres (P GMA-EDMA ) were functionalized with tetraethylenepentamine (TEPA) to generate amino groups which act as a catalyst in hydrolysis of tetraethyl orthosilicate (TEOS) to form Si-containing low molecular weight species. Then the low molecular weight species diffused into the functionalized P GMA-EDMA microspheres by induction force of the amino groups to form polymer/silica hybrid microspheres. Finally, the organic polymer templates were removed by calcination, and the large-porous silica microspheres were obtained. The compositions, morphology, size distribution, specific surface area and pore size distribution of the porous silica microspheres were characterized by infrared analyzer, scanning-electron microscopy, dynamic laser scattering, the mercury intrusion method and thermal gravimetric analysis, respectively. The results show that the agglomeration of the hybrid microspheres can be overcome when the templates were functionalized with TEPA as amination reagent, and the yield of 95.7% of the monodisperse large-porous silica microspheres can be achieved with high concentration of polymer templates. The resulting large-porous silica microspheres were modified with octadecyltrichlorosilane (ODS) and the chromatographic evaluation was performed by separating the proteins and the digest of BSA. The baseline separation of seven kinds of protein standards was achieved, and the column delivered a better performance when separating BSA digests comparing with the commercial one currently available. The high column efficiency and good reproducibility present that the large-porous silica microspheres obtained can be used as a matrix for peptide and protein separation. Copyright © 2016 Elsevier B.V. All rights reserved.
Albumin microspheres as an ocular delivery system for pilocarpine nitrate.
Rathod, Sudha; Deshpande, S G
2008-01-01
Pilocarpine nitrate loaded egg albumin microspheres were prepared by thermal denaturation process in the size range of 1-12 mum. A series of batches were prepared to study factors, which may affect the size and entrapment efficiency of drug in microspheres and optimized the process. Drug loaded microspheres so obtained were evaluated for their size, entrapment efficiency, release rate and biological response. Electron photomicrographs were taken (8000X) to study the morphological characteristics of microspheres. The entrapment and encapsulation of pilocarpine after process optimization was found to be 82.63% and 62.5% respectively. In vitro dissolution rate studies revealed that the release of drug from the microspheres followed spherical matrix mechanism. Biological response of microspheric suspension was measured by reduction in intraocular pressure in albino rabbit eyes and compared with marketed eye drops. Various pharmacokinetic parameters viz. onset of action, duration of action, Tmax and AUC were studied. A measurable difference was found in the mean miotic response, duration and AUC of pilocarpine nitrate microspheric suspension.
Aerosol Generation by Modern Flush Toilets.
Johnson, David; Lynch, Robert; Marshall, Charles; Mead, Kenneth; Hirst, Deborah
A microbe-contaminated toilet will produce bioaerosols when flushed. We assessed toilet plume aerosol from high efficiency (HET), pressure-assisted high efficiency (PAT), and flushometer (FOM) toilets with similar bowl water and flush volumes. Total and droplet nuclei "bioaerosols" were assessed. Monodisperse 0.25-1.9- μ m fluorescent microspheres served as microbe surrogates in separate trials in a mockup 5 m 3 water closet (WC). Bowl water seeding was approximately 10 12 particles/mL. Droplet nuclei were sampled onto 0.2- μ m pore size mixed cellulose ester filters beginning 15 min after the flush using open-face cassettes mounted on the WC walls. Pre- and postflush bowl water concentrations were measured. Filter particle counts were analyzed via fluorescent microscopy. Bowl headspace droplet count size distributions were bimodal and similar for all toilet types and flush conditions, with 95% of droplets < 2 μ m diameter and > 99% < 5 μ m. Up to 145,000 droplets were produced per flush, with the high-energy flushometer producing over three times as many as the lower energy PAT and over 12 times as many as the lowest energy HET despite similar flush volumes. The mean numbers of fluorescent droplet nuclei particles aerosolized and remaining airborne also increased with flush energy. Fluorescent droplet nuclei per flush decreased with increasing particle size. These findings suggest two concurrent aerosolization mechanisms-splashing for large droplets and bubble bursting for the fine droplets that form droplet nuclei.
Viles, C L; Sieracki, M E
1992-01-01
Accurate measurement of the biomass and size distribution of picoplankton cells (0.2 to 2.0 microns) is paramount in characterizing their contribution to the oceanic food web and global biogeochemical cycling. Image-analyzed fluorescence microscopy, usually based on video camera technology, allows detailed measurements of individual cells to be taken. The application of an imaging system employing a cooled, slow-scan charge-coupled device (CCD) camera to automated counting and sizing of individual picoplankton cells from natural marine samples is described. A slow-scan CCD-based camera was compared to a video camera and was superior for detecting and sizing very small, dim particles such as fluorochrome-stained bacteria. Several edge detection methods for accurately measuring picoplankton cells were evaluated. Standard fluorescent microspheres and a Sargasso Sea surface water picoplankton population were used in the evaluation. Global thresholding was inappropriate for these samples. Methods used previously in image analysis of nanoplankton cells (2 to 20 microns) also did not work well with the smaller picoplankton cells. A method combining an edge detector and an adaptive edge strength operator worked best for rapidly generating accurate cell sizes. A complete sample analysis of more than 1,000 cells averages about 50 min and yields size, shape, and fluorescence data for each cell. With this system, the entire size range of picoplankton can be counted and measured. Images PMID:1610183
Hernán Pérez de la Ossa, D; Ligresti, A; Gil-Alegre, M E; Aberturas, M R; Molpeceres, J; Di Marzo, V; Torres Suárez, A I
2012-08-10
Cannabinoids show promise for the treatment of various medical conditions such as emesis, anorexia, pain, cancer, multiple sclerosis, Parkinson's disease and glaucoma. However, their high lipohilicity and instability complicate their handling and dosing, and restrict their use as pharmaceuticals. The objective of the present work was to assess the feasibility of developing cannabinoid loaded poly-ε-caprolactone (PCL) microparticles prepared by the oil-in-water emulsion-solvent evaporation technique as a suitable dosage form for their administration. Spherical microparticles with a size range of 20-50 μm, and high entrapment efficiency (around 100%) were obtained. Cannabidiol (CBD) dissolved in the polymeric matrix of the microspheres was slowly released in vitro within 10 days. In vitro cell viability studies demonstrated the antitumoral activity of CBD released from microparticles. After 4 and 7 days of incubation, CBD in microspheres significantly inhibited the growth of MDA-MB-231 cells by 60% as compared to the 50% attained with free drug. The results suggest that PCL microparticles could be an alternative delivery system for long-term cannabinoid administration, showing potential therapeutic advantages over free drug. Copyright © 2012 Elsevier B.V. All rights reserved.
The osteogenic response of mesenchymal stem cells to an injectable PLGA bone regeneration system.
Curran, Judith M; Fawcett, Sandra; Hamilton, Lloyd; Rhodes, Nicholas P; Rahman, Cheryl V; Alexander, Morgan; Shakesheff, Kevin; Hunt, John A
2013-12-01
The enrichment of substrates/surfaces with selected functional groups, methyl (-CH3), allyl amine (-NH2), allyl alcohol (-OH) and acrylic acid (-COOH), can be used to trigger mesenchymal stem (MSC) cell differentiation into specified lineages, minimising the need for exogenous biological supplementation. We present the successful translation of this research phenomenon to an injectable two phase injectable PLGA system, utilising plasma techniques, for the repair of bone defects. Modified microspheres were characterised using water contact angel (WCA), X-ray Photon Spectroscopy (XPS) and scanning electron microscopy (SEM). When cultured in contact with MSCs in vitro, the ability of the modified particles, within the 2 phase system, to induce differentiation was characterised using quantitative assays for cell viability and histological analysis for key markers of differentiation throughout the entirety of the three dimensional scaffold. Biological analysis proved that selected modified microspheres have the ability to induce MSC osteogenic (-NH2 modified scaffolds) and chondrogenic (-OH modified scaffolds) differentiation throughout the entirety of the formed scaffold. Therefore optimised plasma modification of microspheres is an effective tool for the production of injectable systems for the repair of bone and cartilage defects. Copyright © 2013 Elsevier Ltd. All rights reserved.
NASA Astrophysics Data System (ADS)
Jones, S. K.; Winter, J. G.
2001-02-01
It is known that significant heating can be generated by magnetic hysteresis effects in small ferromagnetic particles exposed to a rapidly alternating magnetic field. If such particles can be made to infiltrate the vascular bed surrounding a tumour by intravascular infusion then it may be possible to generate sufficient heating to destroy the tumour by hyperthermia. One of the constraints on such a technique is the limited amount of magnetic material that can be delivered to a tumour via the intravascular route and the consequent heating that can be induced by this material. Here, we report on a series of experiments in which doses of microspheres containing different amounts of ferromagnetic material were infused into rabbit kidneys via the renal artery with the aim of testing whether adequate tissue heating could be achieved using realistic concentrations of the embolised material. Heating rates were measured for each infused quantity under similar conditions with the animal alive and dead to examine the role of blood flow in the heating process. The results show that tissue temperatures above the therapeutic threshold of 42 °C can be readily achieved using this method with clinically relevant concentrations of microspheres in living tissue.
Microporous polymeric 3D scaffolds templated by the layer-by-layer self-assembly.
Paulraj, Thomas; Feoktistova, Natalia; Velk, Natalia; Uhlig, Katja; Duschl, Claus; Volodkin, Dmitry
2014-08-01
Polymeric scaffolds serve as valuable supports for biological cells since they offer essential features for guiding cellular organization and tissue development. The main challenges for scaffold fabrication are i) to tune an internal structure and ii) to load bio-molecules such as growth factors and control their local concentration and distribution. Here, a new approach for the design of hollow polymeric scaffolds using porous CaCO3 particles (cores) as templates is presented. The cores packed into a microfluidic channel are coated with polymers employing the layer-by-layer (LbL) technique. Subsequent core elimination at mild conditions results in formation of the scaffold composed of interconnected hollow polymer microspheres. The size of the cores determines the feature dimensions and, as a consequence, governs cellular adhesion: for 3T3 fibroblasts an optimal microsphere size is 12 μm. By making use of the carrier properties of the porous CaCO3 cores, the microspheres are loaded with BSA as a model protein. The scaffolds developed here may also be well suited for the localized release of bio-molecules using external triggers such as IR-light. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Tian, Ailin; Qi, Jing; Liu, Yating; Wang, Fengkang; Ito, Yoichiro; Wei, Yun
2013-01-01
Separation of enantiomers still remains a challenge due to their identical physical and chemical properties in a chiral environment, and the research on specific chiral selector along with separation techniques continues to be conducted to resolve individual enantiomers. In our laboratory the promising magnetic chiral microspheres Fe3O4@SiO2@cellulose-2, 3-bis (3, 5-dimethylphenylcarbamate) have been developed to facilitate the resolution using both its magnetic property and chiral recognition ability. In our present studies this magnetic chiral selector was first purified by centrifuge field flow fractionation, and then used to separate benzoin racemate by a chromatographic method. Uniform-sized and masking-impurity-removed magnetic chiral selector was first obtained by field flow fractionation with ethanol through a spiral column mounted on the type-J planetary centrifuge, and using the purified magnetic chiral selector, the final chromatographic separation of benzoin racemate was successfully performed by eluting with ethanol through a coiled tube (wound around the cylindrical magnet to retain the magnetic chiral selector as a stationary phase) submerged in dry ice. In addition, an external magnetic field facilitates the recycling of the magnetic chiral selector. PMID:23891368
NASA Astrophysics Data System (ADS)
Wang, Guangshuo; Zhao, Dexing; Li, Nannan; Wang, Xuehan; Ma, Yingying
2018-06-01
In this study, poly (ε-caprolactone) (PCL) microspheres loading magnetic Fe3O4 nanoparticles and anti-cancer drug of doxorubicin hydrochloride (DOX) were successfully prepared by a modified solvent-evaporation method. The obtained magnetic composite microspheres exhibited dual features of magnetic resonance imaging and controlled drug delivery. The morphology, structure, thermal behavior and magnetic properties of the drug-loaded magnetic microspheres were investigated in detail by SEM, XRD, DSC and SQUID. The obtained composite microspheres showed superparamagnetic behavior and T2-weighted enhancement effect. The drug loading, encapsulation efficiency, releasing behavior and in vitro cytotoxicity of the drug-loaded composite microspheres were systematically investigated. It was found that the values of drug loading and encapsulation efficiency were 36.7% and 25.8%, respectively. The composite microspheres were sensitive to pH and released in a sustained way, and both the release curves under various pH conditions (4.0 and 7.4) were well satisfied with the biphase kinetics function. In addition, the magnetic response of the drug-loaded microspheres was studied and the results showed that the composite microspheres had a good magnetic stability and strong targeting ability.
Eudragit-coated dextran microspheres of 5-fluorouracil for site-specific delivery to colon.
Rai, Gopal; Yadav, Awesh K; Jain, Narendra K; Agrawal, Govind P
2016-01-01
Objective of the present investigation was to prepare and evaluate the potential of enteric coated dextran microspheres for colon targeting of 5-fluorouracil (5-FU). Dextran microspheres were prepared by emulsification-crosslinking method and the formulation variables studied included different molecular weights of dextran, drug:polymer ratio, volume of crosslinking agent, stirring speed and time. Enteric coating (Eudragit S-100) of dextran microspheres was performed by oil-in-oil solvent evaporation method using different coat:core ratios (4:1 or 8:1). Uncoated and coated dextran microspheres were characterized by particle size, surface morphology, entrapment efficiency, DSC, in vitro drug release in the presence of dextranase and 2% rat cecal contents. The release study of 5-FU from coated dextran microspheres was pH dependent. No release was observed at acidic pH; however, the drug was released quickly where Eudragit starts solublizing there was continuous release of drug from the microspheres. Organ distribution study was suggested that coated dextran microspheres retard the release of drug in gastric and intestinal pH environment and released of drug from microspheres in colon due to the degradation of dextran by colonic enzymes.
Microsphere-based gradient implants for osteochondral regeneration: a long-term study in sheep
Mohan, Neethu; Gupta, Vineet; Sridharan, Banu Priya; Mellott, Adam J; Easley, Jeremiah T; Palmer, Ross H; Galbraith, Richard A; Key, Vincent H; Berkland, Cory J; Detamore, Michael S
2015-01-01
Background: The microfracture technique for cartilage repair has limited ability to regenerate hyaline cartilage. Aim: The current study made a direct comparison between microfracture and an osteochondral approach with microsphere-based gradient plugs. Materials & methods: The PLGA-based scaffolds had opposing gradients of chondroitin sulfate and β-tricalcium phosphate. A 1-year repair study in sheep was conducted. Results: The repair tissues in the microfracture were mostly fibrous and had scattered fissures with degenerative changes. Cartilage regenerated with the gradient plugs had equal or superior mechanical properties; had lacunated cells and stable matrix as in hyaline cartilage. Conclusion: This first report of gradient scaffolds in a long-term, large animal, osteochondral defect demonstrated potential for equal or better cartilage repair than microfracture. PMID:26418471
Abadi, Shaivad Shabee Hulhasan; Moin, Afrasim; Veerabhadrappa, Gangadharappa Hosahalli
2016-01-01
Microparticles are polymeric bodies ranging 1-1000 µm that constitute a variety of forms such as microcapsules, microspheres, microcages, microshells, microrods, biosensors microparticles, radiolabeled microparticles, and so forth. This review focuses on general microparticles, mainly microcapsules and microspheres. Nonsteriodal anti-inflammatory drugs (NSAIDs) are one of the mostcommonly prescribed medications in the world. Most of the NSAIDs available have severe side effects. With increased awareness of NSAID-induced gastrointestinal (GI) side effects, safety has become a priority in treatment of arthritis and other inflammatory diseases with NSAIDs. A trend in NSAID development has been to improve therapeutic efficacy while reducing the severity of GI side effects by altering dosage through modified release to optimize drug delivery. One such approach is the use of fabricated microparticles such as microcapsules and microspheres as carriers of drugs. Microparticles provide delivery of macromolecules and micromolecules via different routes and effectively control the release profile of such drugs. Microcapsules and microspheres are compatible with most natural and synthetic polymers and can be used for several routes of administration, including parenteral, oral, nasal, intra-ocular, topical, and the like. Because of greater stability and multiple manufacturing techniques, microspheres and microcapsules are preferred as drug carriers over other colloidal drug delivery systems. Microparticles provide effective protection of the encapsulated agent against degradation by enzymatic activities, controlled and confined delivery of drugs from a few hours to months, and ingenious administration compared to alternative forms of controlled-release parenteral dosages, such as macro-sized implants. This comprehensive overview of fabricated microparticles describes microencapsulation technologies to produce microparticles for targeted therapy of arthritis and other inflammatory diseases which provide constant and prolonged therapeutic effects that reduce dosing frequency and thereby minimize potential adverse effects of NSAIDs such as GI irritation and insufficient patient compliance. The present review describes the latest developments in microparticulate drug delivery systems and the best alternatives for safe and effective microcapsular systems in a controlled manner for the delivery of NSAIDs.
Zhao, Jing; Zeng, Ming; Zheng, Kaiqiang; He, Xinhua; Xie, Minqiang; Fu, Xiaoyi
2017-01-01
Thermoresponsive P(NIPAM-AA)/Fe3O4/SiO2 microspheres with surface holes serving as carriers were prepared using p-Fe3O4/SiO2 microspheres with a thermoresponsive copolymer. The p-Fe3O4/SiO2 microspheres was obtained using a modified Pickering method and chemical etching. The surface pore size of p-Fe3O4/SiO2 microspheres was in the range of 18.3 nm~37.2 nm and the cavity size was approximately 60 nm, which are suitable for loading and transporting biological macromolecules. P(NIPAM-AA) was synthesized inside and outside of the p-Fe3O4/SiO2 microspheres via atom transfer radical polymerization of NIPAM, MBA and AA. The volume phase transition temperature (VPTT) of the specifically designed P(NIPAM-AA)/Fe3O4/SiO2 microspheres was 42.5 °C. The saturation magnetization of P(NIPAM-AA)/Fe3O4/SiO2 microspheres was 72.7 emu/g. The P(NIPAM-AA)/Fe3O4/SiO2 microspheres were used as carriers to study the loading and release behavior of BSA. This microsphere system shows potential for the loading of proteins as a drug delivery platform. PMID:28772770
Microsphere based improved sunscreen formulation of ethylhexyl methoxycinnamate.
Gogna, Deepak; Jain, Sunil K; Yadav, Awesh K; Agrawal, G P
2007-04-01
Polymethylmethacrylate (PMMA) microspheres of ethylhexyl methoxycinnamate (EHM) were prepared by emulsion solvent evaporation method to improve its photostability and effectiveness as sunscreening agent. Process parameters like stirring speed and aqueous polyvinyl alcohol (PVA) concentration were analyzed in order to optimize the formulations. Shape and surface morphology of the microspheres were examined using scanning electron microscopy. Particle size of the microspheres was determined using laser diffraction particle size analyzer. The PMMA microspheres of EHM were incorporated in water-removable cream base. The in vitro drug release of EHM in pH 7.4 was performed using dialysis membrane. Thin layer chromatography was performed to determine photostability of EHM inside the microspheres. The formulations were evaluated for sun protection factor (SPF) and minimum erythema dose (MED) in albino rats. Cream base formulation containing microspheres prepared using EHM:PMMA in ratio of 1:3 (C(3)) showed slowest drug (EHM) release and those prepared with EHM: PMMA in ratio of 1:1 showed fastest release. The cream base formulations containing EHM loaded microspheres had shown better SPF (more than 16.0) as compared to formulation C(d) that contained 3% free EHM as sunscreen agent and showed SPF 4.66. These studies revealed that the incorporation of EHM loaded PMMA microspheres into cream base had greatly increased the efficacy of sunscreen formulation approximately four times. Further, photostability was also shown to be improved in PMMA microspheres.
POE/PLGA composite microspheres: formation and in vitro behavior of double walled microspheres.
Yang, Yi-Yan; Shi, Meng; Goh, Suat-Hong; Moochhala, Shabbir M; Ng, Steve; Heller, Jorge
2003-03-07
The poly(ortho ester) (POE) and poly(D,L-lactide-co-glycolide) 50:50 (PLGA) composite microspheres were fabricated by a water-in-oil-in-water (w/o/w) double emulsion process. The morphology of the composite microspheres varied depending on POE content. When the POE content was 50, 60 or 70% in weight, the double walled microspheres with a dense core of POE and a porous shell of PLGA were formed. The formation of the double walled POE/PLGA microspheres was analysed. Their in vitro degradation behavior was characterized by scanning electron microscopy, gel permeation chromatography, Fourier-transform infrared microscopy and nuclear magnetic resonance spectroscopy (NMR). It was found that compared to the neat POE or PLGA microspheres, distinct degradation mechanism was achieved in the double walled POE/PLGA microspheres system. The degradation of the POE core was accelerated due to the acidic microenvironment produced by the hydrolysis of the outer PLGA layer. The formation of hollow microspheres became pronounced after the first week in vitro. 1H NMR spectra showed that the POE core was completely degraded after 4 weeks. On the other hand, the outer PLGA layer experienced slightly retarded degradation after the POE core disappeared. PLGA in the double walled microspheres kept more than 32% of its initial molecular weight over a period of 7 weeks.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Xu, Ming; Zhang, Wei-De, E-mail: zhangwd@scut.edu.cn
2015-07-15
Bi{sub 2}MoO{sub 6} hierarchical microspheres were successfully prepared through phase transformation from BiOI microspheres with the assistance of sodium citrate under hydrothermal condition. The possible formation mechanism for the conversion of BiOI to Bi{sub 2}MoO{sub 6} is discussed here. After being annealed at 300 °C for 2 h, the obtained Bi{sub 2}MoO{sub 6} microspheres exhibited remarkably enhanced photocatalytic activity towards the degradation of rhodamine B and phenol. The superior catalytic performance can be attributed to its larger surface area and higher crystallinity. In addition, Bi{sub 2}MoO{sub 6} microspheres are stable during the degradation reaction and can be used repeatedly. -more » Graphical abstract: Bi{sub 2}MoO{sub 6} hierarchical microspheres were successfully prepared through a facile partial anion exchange strategy using BiOI microspheres as self-sacrificing templates. The Bi{sub 2}MoO{sub 6} microspheres show high visible light photocatalytic activity. - Highlights: • Bi{sub 2}MoO{sub 6} microspheres were prepared via self-sacrificing template anion exchange. • Sodium citrate-assisted anion exchange for preparation of Bi{sub 2}MoO{sub 6} photocatalyst. • Bi{sub 2}MoO{sub 6} catalysts show high visible light photocatalytic activity.« less
High-density, microsphere-based fiber optic DNA microarrays.
Epstein, Jason R; Leung, Amy P K; Lee, Kyong Hoon; Walt, David R
2003-05-01
A high-density fiber optic DNA microarray has been developed consisting of oligonucleotide-functionalized, 3.1-microm-diameter microspheres randomly distributed on the etched face of an imaging fiber bundle. The fiber bundles are comprised of 6000-50000 fused optical fibers and each fiber terminates with an etched well. The microwell array is capable of housing complementary-sized microspheres, each containing thousands of copies of a unique oligonucleotide probe sequence. The array fabrication process results in random microsphere placement. Determining the position of microspheres in the random array requires an optical encoding scheme. This array platform provides many advantages over other array formats. The microsphere-stock suspension concentration added to the etched fiber can be controlled to provide inherent sensor redundancy. Examining identical microspheres has a beneficial effect on the signal-to-noise ratio. As other sequences of interest are discovered, new microsphere sensing elements can be added to existing microsphere pools and new arrays can be fabricated incorporating the new sequences without altering the existing detection capabilities. These microarrays contain the smallest feature sizes (3 microm) of any DNA array, allowing interrogation of extremely small sample volumes. Reducing the feature size results in higher local target molecule concentrations, creating rapid and highly sensitive assays. The microsphere array platform is also flexible in its applications; research has included DNA-protein interaction profiles, microbial strain differentiation, and non-labeled target interrogation with molecular beacons. Fiber optic microsphere-based DNA microarrays have a simple fabrication protocol enabling their expansion into other applications, such as single cell-based assays.
NASA Astrophysics Data System (ADS)
Saetchnikov, Vladimir A.; Tcherniavskaia, Elina A.; Saetchnikov, Anton V.; Schweiger, Gustav; Ostendorf, Andreas
2014-05-01
Experimental data on detection and identification of variety of biochemical agents, such as proteins, microelements, antibiotic of different generation etc. in both single and multi component solutions under varied in wide range concentration analyzed on the light scattering parameters of whispering gallery mode optical resonance based sensor are represented. Multiplexing on parameters and components has been realized using developed fluidic sensor cell with fixed in adhesive layer dielectric microspheres and data processing. Biochemical component identification has been performed by developed network analysis techniques. Developed approach is demonstrated to be applicable both for single agent and for multi component biochemical analysis. Novel technique based on optical resonance on microring structures, plasmon resonance and identification tools has been developed. To improve a sensitivity of microring structures microspheres fixed by adhesive had been treated previously by gold nanoparticle solution. Another technique used thin film gold layers deposited on the substrate below adhesive. Both biomolecule and nanoparticle injections caused considerable changes of optical resonance spectra. Plasmonic gold layers under optimized thickness also improve parameters of optical resonance spectra. Biochemical component identification has been also performed by developed network analysis techniques both for single and for multi component solution. So advantages of plasmon enhancing optical microcavity resonance with multiparameter identification tools is used for development of a new platform for ultra sensitive label-free biomedical sensor.
EFFECTS OF THE GRAM STAIN ON MICROSPHERES FROM THERMAL POLYAMINO ACIDS1
Fox, Sidney W.; Yuyama, Shuhei
1963-01-01
Fox, Sidney W. (The Florida State University, Tallahassee) and Shuhei Yuyama. Effects of the Gram stain on microspheres from thermal polyamino acids. J. Bacteriol. 85:279–283. 1963.—Microspheres produced from acid proteinoid accept the Gram stain. The stain is negative, but microspheres produced from mixtures containing a sufficient proportion of lysine proteinoid stain positive. Microspheres produced from mixtures containing the appropriate proportions contain individuals which stain positive and others which stain negative. Images PMID:13959050
Kraus, Jr., Robert H.; Zhou, Feng [Los Alamos, NM; Nolan, John P [Santa Fe, NM
2007-06-19
The present invention is directed to processes of separating, analyzing and/or collecting selected species within a target sample by use of magnetic microspheres including magnetic particles, the magnetic microspheres adapted for attachment to a receptor agent that can subsequently bind to selected species within the target sample. The magnetic microspheres can be sorted into a number of distinct populations, each population with a specific range of magnetic moments and different receptor agents can be attached to each distinct population of magnetic microsphere.
A technique for studying cardiac myosin dynamics using optical tweezers
NASA Astrophysics Data System (ADS)
Paolino, Michael; Migirditch, Sam; Nesmelov, Yuri; Hester, Brooke; Appalachian State Biophysics; Optical Sciences Facility Team
A primary protein involved in human muscle contraction is myosin, which exists in α- and β- isoforms. Myosin exerts forces on actin filaments when ATP is present, driving muscle contraction. A significant decrease in the population of cardiac α-myosin has been linked to heart failure. It is proposed that slow β-myosin in a failing heart could, through introduction of a drug, be made to mimic the action of α-myosin, thereby improving cardiac muscle performance. In working towards testing this hypothesis, the focus of this work is to develop a technique to measure forces exerted by myosin on actin using optical tweezers. An actin-myosin arrangement is constructed between two optically trapped polystyrene microspheres. The displacement of a microsphere is monitored when ATP is introduced, and the force responsible is measured. With this achieved, we can then modify the actin-myosin arrangement, for example with varying amounts of α- and β- myosin and test the effects on forces exerted. In this work, assemblies of actin and myosin molecules and preliminary force measurements are discussed. North Carolina Space Grant.
NASA Astrophysics Data System (ADS)
Tang, Yihao; Zhan, Shuai; Wang, Li; Zhang, Bin; Ding, Minghui
The pure anatase TiO2 hollow microspheres are synthesized by a one-step template-free hydrothermal route. By defining temperature and time limits, we produce TiO2 hollow microspheres with a fluoride-mediated self-transformation. The surface morphology of TiO2 hollow microspheres was studied by SEM. The hollow microspheres have diameters of about 800 nm and are remarkably uniform. The UV-light photocatalytic activity and the stability/multifunction of TiO2 hollow microspheres structure were evaluated by photocatalytic degradation of methylene blue and photocatalytic hydrogen evolution. The excellent photocatalytic activity is attributed to large specific surface area, more active sites, unique hollow structures, and improved light scattering.
Method of detecting luminescent target ions with modified magnetic microspheres
Shkrob, Ilya A; Kaminski, Michael D
2014-05-13
This invention provides methods of using modified magnetic microspheres to extract target ions from a sample in order to detect their presence in a microfluidic environment. In one or more embodiments, the microspheres are modified with molecules on the surface that allow the target ions in the sample to form complexes with specific ligand molecules on the microsphere surface. In one or more embodiments, the microspheres are modified with molecules that sequester the target ions from the sample, but specific ligand molecules in solution subsequently re-extract the target ions from the microspheres into the solution, where the complexes form independent of the microsphere surface. Once the complexes form, they are exposed to an excitation wavelength light source suitable for exciting the target ion to emit a luminescent signal pattern. Detection of the luminescent signal pattern allows for determination of the presence of the target ions in the sample.
Yan, Rui; Zhang, Yaoyao; Wang, Xiaohui; Xu, Jianxiong; Wang, Da; Zhang, Wangqing
2012-02-15
Synthesis of porous poly(styrene-co-acrylic acid) (PS-co-PAA) microspheres through one-step soap-free emulsion polymerization is reported. Various porous PS-co-PAA microspheres with the particle size ranging from 150 to 240 nm and with the pore size ranging from 4 to 25 nm are fabricated. The porous structure of the microspheres is confirmed by the transmission electron microscopy measurement and Brunauer-Emmett-Teller (BET) analysis. The reason for synthesis of the porous PS-co-PAA microspheres is discussed, and the phase separation between the encapsulated hydrophilic poly(acrylic acid) segment and the hydrophobic polystyrene domain within the PS-co-PAA microspheres is ascribed to the pore formation. The present synthesis of the porous PS-co-PAA microspheres is anticipated to be a new and convenient way to fabricate porous polymeric particles. Copyright © 2011 Elsevier Inc. All rights reserved.
Farooq, Umar; Khan, Samiullah; Nawaz, Shahid; Ranjha, Nazar Mohammad; Haider, Malik Salman; Khan, Muhammad Muzamil; Dar, Eshwa; Nawaz, Ahmad
2017-01-01
Abstract Eudragit E 100 and polycaprolactone (PCL) floating microspheres for enhanced gastric retention and drug release were successfully prepared by oil in water solvent evaporation method. Metronidazole benzoate, an anti-protozoal drug, was used as a model drug. Polyvinyl alcohol was used as an emulsifier. The prepared microspheres were observed for % recovery, % degree of hydration, % water uptake, % drug loading, % buoyancy and % drug release. The physico-chemical properties of the microspheres were studied by calculating encapsulation efficiency of microspheres and drug release kinetics. Drug release characteristics of microspheres were studied in simulated gastric fluid and simulated intestinal fluid i.e., at pH 1.2 and 7.4 respectively. Fourier transform infrared spectroscopy was used to reveal the chemical interaction between drug and polymers. Scanning electron microscopy was conducted to study the morphology of the synthesized microspheres. PMID:29491813
Monodisperse, polymeric microspheres produced by irradiation of slowly thawing frozen drops
NASA Technical Reports Server (NTRS)
Rhim, Won-Kyu (Inventor); Hyson, Michael T. (Inventor); Chung, Sang-Kun (Inventor); Colvin, Michael S. (Inventor); Chang, Manchium (Inventor)
1991-01-01
Monodisperse, polymeric microspheres are formed by injecting uniformly shaped droplets of radiation polymerizable monomers, preferably a biocompatible monomer, having covalent binding sites such as hydroxyethylmethacrylate, into a zone, impressing a like charge on the droplet so that they mutually repel each other, spheroidizing the droplets within the zone and collecting the droplets in a pool of cryogenic liquid. As the droplets enter the liquid, they freeze into solid, glassy microspheres, which vaporizes a portion of the cryogenic liquid to form a layer. The like-charged microspheres, suspended within the layer, move to the edge of the vessel holding the pool, are discharged, fall and are collected. The collected microspheres are irradiated while frozen in the cryogenic liquid to form latent free radicals. The frozen microspheres are then slowly thawed to activate the free radicals which polymerize the monomer to form evenly-sized, evenly-shaped, monodisperse polymeric microspheres.
Mesoporous metal oxide microsphere electrode compositions and their methods of making
Parans Paranthaman, Mariappan; Bi, Zhonghe; Bridges, Craig A; Brown, Gilbert M
2014-12-16
Compositions and methods of making are provided for treated mesoporous metal oxide microspheres electrodes. The compositions comprise (a) microspheres with an average diameter between 200 nanometers (nm) and 10 micrometers (.mu.m); (b) mesopores on the surface and interior of the microspheres, wherein the mesopores have an average diameter between 1 nm and 50 nm and the microspheres have a surface area between 50 m.sup.2/g and 500 m.sup.2/g, and wherein the composition has an electrical conductivity of at least 1.times.10.sup.-7 S/cm at 25.degree. C. and 60 MPa. The methods of making comprise forming a mesoporous metal oxide microsphere composition and treating the mesoporous metal oxide microspheres by at least one method selected from the group consisting of: (i) annealing in a reducing atmosphere, (ii) doping with an aliovalent element, and (iii) coating with a coating composition.
Method for the production of fabricated hollow microspheroids
Wickramanayake, Shan; Luebke, David R.
2015-06-09
The method relates to the fabrication of a polymer microspheres comprised of an asymmetric layer surrounding a hollow interior. The fabricated hollow microsphere is generated from a nascent hollow microsphere comprised of an inner core of core fluid surrounded by a dope layer of polymer dope, where the thickness of the dope layer is at least 10% and less than 50% of the diameter of the inner core. The nascent hollow microsphere is exposed to a gaseous environment, generating a vitrified hollow microsphere, which is subsequently immersed in a coagulation bath. Solvent exchange produces a fabricated hollow microsphere comprised of a densified outer skin surrounding a macroporous inner layer, which surrounds a hollow interior. In an embodiment, the polymer is a polyimide or a polyamide-imide, and the non-solvent in the core fluid and the coagulation bath is water. The fabricated hollow microspheres are particularly suited as solvent supports for gas separation processes.