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Sample records for simple fluorescence based

  1. Medically Relevant Assays with a Simple Smartphone and Tablet Based Fluorescence Detection System

    PubMed Central

    Wargocki, Piotr; Deng, Wei; Anwer, Ayad G.; Goldys, Ewa M.

    2015-01-01

    Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera) and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 g/mL and 0.938 g in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis. PMID:26007723

  2. Medically relevant assays with a simple smartphone and tablet based fluorescence detection system.

    PubMed

    Wargocki, Piotr; Deng, Wei; Anwer, Ayad G; Goldys, Ewa M

    2015-01-01

    Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera) and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 g/mL and 0.938 g in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis. PMID:26007723

  3. A simple levulinate-based ratiometric fluorescent probe for sulfite with a large emission shift.

    PubMed

    Liu, Caiyun; Wu, Huifang; Yang, Wen; Zhang, Xiaoling

    2014-01-01

    A simple 4-hydroxynaphthalimide-derived colorimetric and ratiometric fluorescent probe (1) containing a receptor of levulinate moiety was designed and synthesized to monitor sulfite. Probe 1 could quantificationally detect sulfite by a ratiometric fluorescence spectroscopy method with high selectivity and sensitivity. Specially, probe 1 exhibited a 100 nm red-shifted absorption spectrum along with the color changes from colorless to yellow, and 103 nm red-shifted emission spectra upon the addition of sulfite. Thus, 1 can serve as a "naked-eye" probe for sulfite. Further, the recognition mechanism of probe 1 for sulfite was confirmed using nuclear magnetic resonance and electrospray ionization mass spectrometry. Also, the preliminary practical application demonstrated that our proposed probe provided a promising method for the determination of sulfite. PMID:24813958

  4. Simple Adhesive-Tape-Based Sampling of Tomato Surfaces Combined with Rapid Fluorescence In Situ Hybridization for Salmonella Detection▿

    PubMed Central

    Bisha, Bledar; Brehm-Stecher, Byron F.

    2009-01-01

    A simple adhesive-tape-based method for sampling of tomato surfaces was combined with fluorescence in situ hybridization for rapid culture-independent detection of Salmonella strains. Tapes could also be placed face-down on selective agar for on-tape enrichment of captured Salmonella cells. Overlay of cell-charged tapes with small volumes of liquid enrichment media enabled subsequent detection of tape-captured Salmonella via flow cytometry. PMID:19124588

  5. A simple and sensitive label-free fluorescent approach for protein detection based on a Perylene probe and aptamer.

    PubMed

    Lv, Zhenzhen; Liu, Jinchuan; Bai, Wenhui; Yang, Shuming; Chen, Ailiang

    2015-02-15

    Highly sensitive detection of proteins is of great importance for effective clinical diagnosis and biomedical research. However, so far most detection methods rely on antibody-based immunoassays and are usually laborious and time-consuming with poor sensitivity. Here, we developed a simple and ultra-sensitive method to detect a biomarker protein-thrombin by taking advantage of the fluorescent probe Perylene tetracarboxylic acid diimide (PTCDI) derivatives and thrombin aptamer. The water-soluble dye PTCDI shows strong fluorescence in buffer solution for the existence of free dye monomer, but becomes weak after aggregation through self-assembly on nucleic acid aptamer. In the presence of thrombin, it specifically binds to thrombin aptamer which causes the conformational transition between aptamer and PTCDI and results in a significant fluorescence recovery. The results showed that as low as 40 pM of thrombin could be detected by this method. The high sensitivity of the developed sensing system mainly attributes to the ultra-sensitivity of the fluorescence intensity changes of PTCDI. With the specificity of aptamer, the assay exhibited high selectivity for thrombin against three other proteins (bovine serum albumin, lysozyme, mouse IgG) and 1% diluted fetal bovine serum. The detection method might be extended to sensitive detection of a variety of proteins for its advantages of isothermal conditions required, simple and rapid without multiple separation and washing steps. PMID:25310484

  6. Development and validation of a simple cell-based fluorescence assay for dipeptidyl peptidase 1 (DPP1) activity.

    PubMed

    Thong, Bob; Pilling, James; Ainscow, Edward; Beri, Raj; Unitt, John

    2011-01-01

    Dipeptidyl peptidase 1 (DPP1) (EC 3.4.14.1; also known as cathepsin C, cathepsin J, dipeptidyl aminopeptidase, and dipeptidyl aminotransferase) is a lysosomal cysteinyl protease of the papain family involved in the intracellular degradation of proteins. Isolated enzyme assays for DPP1 activity using a variety of synthetic substrates such as dipeptide or peptide linked to amino-methyl-coumarin (AMC) or other fluorophores are well established. There is, however, no report of a simple whole-cell-based assay for measuring lysosomal DPP1 activity other than the use of flow cytometry (fluorescence-activated cell sorting) or the use of invasive activity-based probes or the production of physiological products such as neutrophil elastase. The authors investigated a number of DPP1 fluorogenic substrates that have the potential to access the lysosome and enable the measurement of DPP1 enzyme activity in situ. They describe the development and evaluation of a simple noninvasive fluorescence assay for measuring DPP1 activity in fresh or cryopreserved human THP-1 cells using the substrate H-Gly-Phe-AFC (amino-fluoro-coumarin). This cell-based fluorescence assay can be performed in a 96-well plate format and is ideally suited for determining the cell potency of potential DPP1 enzyme inhibitors. PMID:21088147

  7. A simple and sensitive fluorescent sensor for methyl parathion based on L-tyrosine methyl ester functionalized carbon dots.

    PubMed

    Hou, Juying; Dong, Jing; Zhu, Haishuang; Teng, Xue; Ai, Shiyun; Mang, Minglin

    2015-06-15

    In this paper, a simple and sensitive fluorescent sensor for methyl parathion is developed based on L-tyrosine methyl ester functionalized carbon dots (Tyr-CDs) and tyrosinase system. The carbon dots are obtained by simple hydrothermal reaction using citric acid as carbon resource and L-tyrosine methyl ester as modification reagent. The carbon dots are characterized by transmission electron microscope, high resolution transmission electron microscopy, X-ray diffraction spectrum, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. The carbon dots show strong and stable photoluminescence with a quantum yield of 3.8%. Tyrosinase can catalyze the oxidation of tyrosine methyl ester on the surface of carbon dots to corresponding quinone products, which can quench the fluorescence of carbon dots. When organophosphorus pesticides (OPs) are introduced in system, they can decrease the enzyme activity, thus decrease the fluorescence quenching rate. Methyl parathion, as a model of OPs, was detected. Experimental results show that the enzyme inhibition rate is proportional to the logarithm of the methyl parathion concentration in the range 1.010(-10)-1.010(-4) M with the detection limit (S/N=3) of 4.810(-11) M. This determination method shows a low detection limit, wide linear range, good selectivity and high reproducibility. This sensing system has been successfully used for the analysis of cabbage, milk and fruit juice samples. PMID:25558870

  8. A Simple Fluorescence Probe Based on Aggregation-Induced Emission (AIE) Property for the Detection of Mg(2+) Ions.

    PubMed

    Bian, Yan-Jiang; Wang, Lu-Qiong; Cao, Fu-Xiang; Tang, Li-Jun

    2016-01-01

    A simple aggregation-induced emission-based fluorescence probe (1) for Mg(2+) was synthesized by condensation of benzene-1, 2-diamine with 5-bromo-2-hydroxybenzaldehyde, This compound shows favourable character of the AIE-active molecules. More importantly, after addition of Mg(2+) to probe (1) in acetonitrile, the solution changed from colorless to yellow colour solution under ultraviolet (UV) radiation obtained from hand-held UV lamp, this finding suggested that probe (1) can be used to detect Mg(2+) by colorimetric detection. Detection limit can reach 2.31נ10(-5) M(-1). The practical value of the selective and sensitive fluorescence indicators was confirmed by its application to detection of magnesium ion in acetonitrile. PMID:26547420

  9. Dual-channel detection of Cu2+ and F- with a simple Schiff-based colorimetric and fluorescent sensor

    NASA Astrophysics Data System (ADS)

    Na, Yu Jeong; Choi, Ye Won; Yun, Jin Yeong; Park, Kyung-Min; Chang, Pahn-Shick; Kim, Cheal

    2015-02-01

    A simple and easily synthesized colorimetric and fluorescent receptor 1, based on 4-diethylaminosalicylaldehyde moieties as a binding and signaling unit, has been synthesized and characterized. The receptor 1 has a selective colorimetric sensing ability for copper (II) ion by changing color from colorless to yellow in aqueous solution, and could be utilized to monitor Cu(II) over a wide pH range of 4-11. In addition, the detection limit (12 ?M) of 1 for Cu2+ is much lower than that (30 ?M) recommended by WHO in drinking water, and its copper complex could be reversible simply through treatment with a proper reagent such as EDTA. Moreover, receptor 1 exhibited both a color change from colorless to yellow and fluorescence enhancement with a red shift upon addition to F- in DMSO. The recognition mechanism was attributed to the intermolecular proton transfer between the hydroxyl group of the receptor and the fluoride.

  10. Simple and sensitive fluorescent and electrochemical trinitrotoluene sensors based on aqueous carbon dots.

    PubMed

    Zhang, Lingling; Han, Yujie; Zhu, Jinbo; Zhai, Yanling; Dong, Shaojun

    2015-02-17

    Aqueous N-rich carbon dots (CDs), prepared by the microwave-assisted pyrolysis method, are applied as a dual sensing platform for both the fluorescent and electrochemical detection of 2,4,6-trinitrotoluene (TNT). The fluorescent sensing platform is established on the strong TNT-amino interaction which can quench the photoluminescence of amino functionalized CDs through charge transfer. The resultant linear detection ranges from 10 nM to 1.5 ?M with a fast response time of 30 s. Glassy carbon electrode modified with CDs exhibits a fine capability for TNT reduction with the linear range from 5 nM to 30 ?M, better than that obtained by the fluorescent method. Moreover, the minimum distinguishable response concentration with respect to these two methods is down to the nanomolar level with a high specificity and sensitivity. PMID:25600090

  11. A novel and simple fluorescent and colorimetric primary chemosensor based on Congo-Red for sulfite and resultant complex as secondary fluorescent chemosensor towards carbonate ions: Fluorescent probe mimicking INHIBIT logic gate.

    PubMed

    Tavallali, Hossein; Deilamy-Rad, Gohar; Parhami, Abolfath; Lohrasbi, Sajedeh

    2016-03-01

    A simple receptor based on Congo-Red (CR) was prepared by complexation of CR into two equivalents of Cu (II) ([CR-(Cu)2]) and it has been designed for detection of sulfite and carbonate ions. This chemosensor exhibits high sensitivity for sulfite over other anions in aqueous buffer solution. It exhibits colorimetric 'naked eye' and fluorometric responses to SO3(2-) which results from the addition of SO3(2)(-) to CR diazo moiety. Hereupon, CO3(2-) greatly limits the fluorescence of the resultant sulfite-receptor complex via a hydrogen bonding interaction ([CR-(Cu)2]-SO3). This system can be applied for selective detection of CO3(2-) in the presence of other anions. The detection limits of SO3(2-), calculated by the colorimetric and fluorometric methods, were found to be 0.07 and 0.09µmolL(-)(1), respectively. The sulfite-receptor complex also displayed the ability to detect up to 0.06µmolL(-)(1) CO3(2-). The fluorescence output mimicked 'INHIBIT' logic gate function. The output was exhibited by the intramolecular charge transfer of the [CR-(Cu)2] probe, and was provided by chemical inputs (SO3(2-) and CO3(2-)). PMID:26717828

  12. Heterogeneous Solvatochromism of Fluorescent DNA-Stabilized Silver Clusters Precludes Use of Simple Onsager-Based Stokes Shift Models.

    PubMed

    Copp, Stacy M; Faris, Alexis; Swasey, Steven M; Gwinn, Elisabeth G

    2016-02-18

    The diverse optical and chemical properties of DNA-stabilized silver clusters (AgN-DNAs) have challenged the development of a common model for these sequence-tunable fluorophores. Although correlations between cluster geometry and fluorescence color have begun to shed light on how the optical properties of AgN-DNAs are selected, the exact mechanisms responsible for fluorescence remain unknown. To explore these mechanisms, we study four distinct purified AgN-DNAs in ethanol-water and methanol-water mixtures and find that the solvatochromic behavior of AgN-DNAs varies widely among different cluster species and differs markedly from prior results on impure material. Placing AgN-DNAs within the context of standard Lippert-Mataga solvatochromism models based on the Onsager reaction field, we show that such nonspecific solvent models are not universally applicable to AgN-DNAs. Instead, alcohol-induced solvatochromism of AgN-DNAs may be governed by changes in hydration of the DNA template, with spectral shifts resulting from cluster shape changes and/or dielectric changes in the local vicinity of the cluster. PMID:26831218

  13. Simple and Sensitive Molecularly Imprinted Polymer - Mn-Doped ZnS Quantum Dots Based Fluorescence Probe for Cocaine and Metabolites Determination in Urine.

    PubMed

    Chantada-Vázquez, María Pilar; Sánchez-González, Juan; Peña-Vázquez, Elena; Tabernero, María Jesús; Bermejo, Ana María; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

    2016-03-01

    A new molecularly imprinted polymer (MIP)-based fluorescent artificial receptor has been prepared by anchoring a selective MIP for cocaine (COC) on the surface of polyethylene glycol (PEG) modified Mn-doped ZnS quantum dots (QDs). The prepared material combines the high selectivity attributed to MIPs and the sensitive fluorescent property of the Mn-doped ZnS QDs. Simple and low cost methods have therefore been optimized for assessing cocaine abuse in urine by monitoring the fluorescence quenching when the template (COC) and also metabolites from COC [benzoylecgonine (BZE) and ecgonine methyl ester (EME)] are present. Fluorescence quenching was not observed when performing experiments with other drugs of abuse (and their metabolites) or when using nonimprinted polymer (NIP)-coated QDs. Under optimized operating conditions (1.5 mL of 200 mg L(-1) MIP-coated QDs solution, pH 5.5, and 15 min before fluorescence scanning) two analytical methods were developed/validated. One of the procedures (direct method) consisted of urine sample 1:20 dilution before fluorescence measurements. The method has been found to be fast, precise, and accurate, but the standard addition technique for performing the analysis was required because of the existence of matrix effect. The second procedure performed a solid phase extraction (SPE) first, avoiding matrix effect and allowing external calibration. The limits of detection of the methods were 0.076 mg L(-1) (direct method) and 0.0042 mg L(-1) (SPE based method), which are lower than the cutoff values for confirmative conclusions regarding cocaine abuse. PMID:26857857

  14. Simple pyridyl-salicylimine-based fluorescence "turn-on" sensors for distinct detections of Zn2+, Al3+ and OH- ions in mixed aqueous media.

    PubMed

    Shellaiah, Muthaiah; Wu, Yen-Hsing; Lin, Hong-Cheu

    2013-05-21

    Simple pyridyl-salicylimine derivatives (F1, F2 and F3) are reported for the first time as fluorescence "turn-on" sensors for distinct detections of Zn(2+), Al(3+) and OH(-) ions in mixed-aqueous media CH3CN/H2O with volume ratios of 6/4 and 3/7 (at pH = 7 and 25 C) via internal charge transfer (ICT), chelation enhanced fluorescence (CHEF), and deprotonation mechanisms. F1 and F2 show diverse turn-on sensing applications to Zn(2+), Al(3+) and OH(-) ions, but F3 exhibited the fluorescence turn-on sensing to Al(3+) and OH(-) ions in CH3CN/H2O (6/4; vol/vol). F1+Zn(2+) and F2+Zn(2+) complexes revealed the reversibilities and ratiometric displacements of Zn(2+) with ethylene diamine tetra acetic acid (EDTA) and Al(3+) ions, respectively, in CH3CN/H2O (6/4; vol/vol). On the other hand, F1, F2 and F3 in CH3CN/H2O (3/7; vol/vol) showed sensitivities only to Al(3+) ions but negligible selectivities to OH(-) ions. Stoichiometry of all sensor complexes were calculated as 1 : 1 by job's plots based on UV/Vis and PL titrations. The complex formation and binding sites of all sensor materials were well characterized by (1)H, (13)C NMR, and mass (FAB) spectral analysis. Detection limits were calculated from standard deviations and linear fitting calculations. The association constant (log K(a)) values of sensor complexes were evaluated from the fluorescence binding isotherms. The fluorescence decay constant (?) values were estimated from time resolved fluorescence studies. Time, temperature, pH and solvent concentration effects towards sensor responses were fully investigated in this report. PMID:23531941

  15. A highly selective colorimetric and fluorescent chemosensor for Al(III) based-on simple naphthol in aqueous solution.

    PubMed

    Liu, Zhaodi; Xu, Huajie; Sheng, Liangquan; Chen, Shuisheng; Huang, Deqian; Liu, Jie

    2016-03-15

    A colorimetric and fluorescent chemosensor (L) for Al(III) was synthesized and fully characterized. L could be both used as a colorimetric and fluorescent chemosensor for the detection of Al(3+) ions with low detection limit (8.87×10(-7)M) in CH3CN-H2O (1:1, v/v) solution. The binding ratio of L-Al(3+) was determined from the Job plot (absorption and fluorescence spectra) and MALDI-TOF MS data to be 1:1. The binding constant (Ka) of Al(3+) binding to L was calculated to be 4.8×10(5)M(-1) from a Benesi-Hildebrand plot. Moreover, the binding site of L with Al(3+) was determined by (1)H NMR titration experiment. PMID:26690670

  16. A highly selective colorimetric and fluorescent chemosensor for Al(III) based-on simple naphthol in aqueous solution

    NASA Astrophysics Data System (ADS)

    Liu, Zhaodi; Xu, Huajie; Sheng, Liangquan; Chen, Shuisheng; Huang, Deqian; Liu, Jie

    2016-03-01

    A colorimetric and fluorescent chemosensor (L) for Al(III) was synthesized and fully characterized. L could be both used as a colorimetric and fluorescent chemosensor for the detection of Al3 + ions with low detection limit (8.87 × 10- 7 M) in CH3CN-H2O (1:1, v/v) solution. The binding ratio of L-Al3 + was determined from the Job plot (absorption and fluorescence spectra) and MALDI-TOF MS data to be 1:1. The binding constant (Ka) of Al3 + binding to L was calculated to be 4.8 × 105 M- 1 from a Benesi-Hildebrand plot. Moreover, the binding site of L with Al3 + was determined by 1H NMR titration experiment.

  17. Selective fluorescence sensing of Cu(II) and Zn(II) using a simple Schiff base ligand: Naked eye detection and elucidation of photoinduced electron transfer (PET) mechanism

    NASA Astrophysics Data System (ADS)

    Ganguly, Aniruddha; Ghosh, Soumen; Kar, Samiran; Guchhait, Nikhil

    2015-05-01

    A simple Schiff base compound 2-((cyclohexylmethylimino)-methyl)-naphthalen-1-ol (2CMIMN1O) has been synthesized and characterized by 1H NMR, 13C NMR and FT-IR spectroscopic techniques. A significantly low emission yield of the compound has been rationalized in anticipation with photo-induced electron transfer (PET) from the imine receptor moiety to the naphthalene fluorophore unit. Consequently, an evaluation of the transition metal ion-induced modification of the fluorophore-receptor communication reveals the promising prospect of the title compound to function as a chemosensor for Cu2+ and Zn2+ ions selectively, through remarkable fluorescence enhancement as well as visual changes. While perturbation of the PET process has been argued to be the plausible mechanism behind the fluorescence enhancement, the selectivity for these two metal ions has been interpreted on the grounds of an appreciably strong binding interaction. Particularly notable aspects regarding the chemosensory activity of the compound is its ability to detect the aforesaid transition metal ions down to the level of micromolar concentration (detection limit being 2.74 and 2.27 ppm respectively), along with a simple and efficient synthetic procedure.

  18. Rapid and simple quantitation of methamphetamine by using a homogeneous time-resolved fluoroimmunoassay based on fluorescence resonance energy transfer from europium to Cy5.

    PubMed

    Kimura, Hiroko; Matsumoto, Kazuko; Mukaida, Masahiro

    2005-01-01

    A rapid and simple homogeneous time-resolved fluoroimmunoassay based on fluorescence resonance energy transfer from europium (Eu) to cyanine dye (Cy5) has been developed for the quantitation of methamphetamine. In this assay, Eu chelate was labeled to a conjugate of methamphetamine and bovine serum albumin (MA-BSA), as an energy donor, and Cy5 was labeled to anti-MA as an energy acceptor. The close proximity between the two labels in the immunocomplex permits energy transfer from the excited Eu(3+) donor. Therefore, by measuring the sensitized emission of Cy5 with the time-resolved assay, immunocomplex of MA-BSA and anti-MA can be measured in the homogeneous solution without separation steps within 30 min. By a competitive immunoassay, MA could be assayed in the range 0.1-1,000 ng/mL. The intra-assay variations were 5.4-14.8% at 5 different concentrations. When urine or serum samples were examined, the quenching of Eu fluorescence was observed, but the acceptor-to-donor ratio constantly depended upon the dilution of samples. Twenty urine samples were assayed, and the data showed a good correlation to those obtained by gas chromatography (r = 0.94). The homogeneous assay using Eu-Cy5 energy transfer is time-saving without any washing procedures and is suitable for screening drugs that are commonly abused. PMID:16356338

  19. Simple time-saving method for iron determination based on fluorescence quenching of an azaflavanon-3-ol compound.

    PubMed

    Başoğlu, Aysel; Tosun, Gonca; Ocak, Miraç; Alp, Hakan; Yaylı, Nurettin; Ocak, Ümmühan

    2015-03-18

    A simple and time-saving spectrofluorometric method developed using an azaflavanon-3-ol compound was used for the determination of iron in various food samples. Nitric acid and hydrogen peroxide were used for digestion of samples in a closed microwave system. The method was validated by analyzing two certified reference materials (CRM-SA-C Sandy Soil C and Mixed Polish Herbs INCT-MPH-2). Measurements were carried out using a modified standard addition method. The standard addition graph was linear until 21.6 mg/L in the determination of iron(III). Detection and quantification limits were 0.81 and 2.4 mg/L, respectively. Satisfactory accuracy was obtained for spinach, dill, mint, purslane, rocket, red lentils, dry beans, and two iron medicinal tablets. High recoveries were found for streamwater samples fortified at three different concentrations. The method is simple, time-saving, cost-effective, and suitable for the determination of the iron content of foods. PMID:25723252

  20. A simple and sensitive HPLC method based on pre-column fluorescence labelling for multiple classes of plant growth regulator determination in food samples.

    PubMed

    Li, Guoliang; Liu, Shucheng; Sun, Zhiwei; Xia, Lian; Chen, Guang; You, Jinmao

    2015-03-01

    The determination of trace plant growth regulator (PGR) has received more and more attentions in the field of phytophysiology and food safety. But the simple and sensitive method for simultaneously analysing multiple classes of PGR remains poorly investigated. In this study, a new pre-column fluorescence labelling method using 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methylbenzenesulfonate (BCETS) as the labelling reagent has been developed for simultaneous determination of seven PGRs (i.e., indole-3-acetic acid, 3-indolybutyric acid, 3-indolepropionic acid, jasmonic acid, gibberellin A3, 1-naphthylacetic acid and 2-naphthaleneacetic acid) by HPLC with fluorescent detection (FLD). The proposed method offered the LOD of 0.34-0.73 ng/mL for seven PGRs, which were significantly lower than the reported methods. The crude extract without complex pre-treatments and purification was directly labelled by BCETS and analysed by HPLC-FLD, which facilitates the high-throughput sample screening. This method was proven to be inexpensive, simple, selective, sensitive, accurate and reliable for trace PGR determination. PMID:25306326

  1. Genetic diversity analysis of sugarcane germplasm based on fluorescence-labeled simple sequence repeat markers and a capillary electrophoresis-based genotyping platform

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetic diversity analysis, which refers to the elaboration of total extent of genetic characteristics in the genetic makeup of a certain species, constitutes a classical strategy for the study of diversity, population genetic structure, and breeding practices. In this study, fluorescence-labeled se...

  2. A highly selective and simple fluorescent sensor for mercury (II) ion detection based on cysteamine-capped CdTe quantum dots synthesized by the reflux method.

    PubMed

    Ding, Xiaojie; Qu, Lingbo; Yang, Ran; Zhou, Yuchen; Li, Jianjun

    2015-06-01

    Cysteamine (CA)-capped CdTe quantum dots (QDs) (CA-CdTe QDs) were prepared by the reflux method and utilized as an efficient nano-sized fluorescent sensor to detect mercury (II) ions (Hg(2+) ). Under optimum conditions, the fluorescence quenching effect of CA-CdTe QDs was linear at Hg(2+) concentrations in the range of 6.0-450?nmol/L. The detection limit was calculated to be 4.0?nmol/L according to the 3? IUPAC criteria. The influence of 10-fold Pb(2+) , Cu(2+) and Ag(+) on the determination of Hg(2+) was?based on crude QDs). Furthermore, the detection sensitivity and selectivity were much improved relative to a sensor based on the CA-CdTe QDs probe, which was prepared using a one-pot synthetic method. This CA-CdTe QDs sensor system represents a new feasibility to improve the detection performance of a QDs sensor by changing the synthesis method. PMID:25263990

  3. A simple but efficient strategy to enhance hydrostability of intensely fluorescent Mg-based coordination polymer (CP) via forming a composite of CP with hydrophobic PVDF.

    PubMed

    Zhai, Lu; Zhang, Wen-Wei; Zuo, Jing-Lin; Ren, Xiao-Ming

    2016-02-16

    A coordination polymer (CP) of Mg(2+) with 1,3,5-benzenetricarboxylate (BTC(3-)) was synthesized using a solvothermal method. The Mg-CP, with a formula of Mg3(BTC)(HCOO)3(DMF)3, crystallizes in the trigonal space group P3[combining macron], with cell parameters of a = b = 13.972(5) , c = 8.090(5) and V = 1367.6(11) (3), and shows a lamella structure built from planar rosette-type hexanuclear architectures. The Mg-CP emits intense blue fluorescence arising from ?* ? ? transition of intra-ligand of BTC(3-) with 21.69% quantum yield, yet it exhibits poor stability to water. The composites of Mg-CP with hydrophobic polyvinylidene fluoride (PVDF) were sequentially prepared by mechanically mixed, tableted and annealed processes, which showed good compatibility between Mg-CP and PVDF, high hydrostability, and intense blue emission. This study suggests a simple but efficient method to solve the drawbacks of some functional CPs unstable to water and to promote them as practical applications in the field of functional materials. PMID:26790523

  4. Highly selective detection of Cr(VI) in water matrix by a simple 1,8-naphthalimide-based turn-on fluorescent sensor.

    PubMed

    Zhang, Zhiyuan; Sha, Chunming; Liu, Aifeng; Zhang, Zhenyu; Xu, Dongmei

    2015-03-01

    4-Bis-(2-hydroxyethyl)amino-N-n-butyl-1,8-naphthalimide was developed into a turn-on fluorescent sensor for Cr(VI). In the mixture of N,N-dimethylformamide and deionized water (1:9, v/v), the sensor could highly selectively distinguish CrO4(2-) from Cr(3+), Na(+), K(+), Mg(2+), Ca(2+), Mn(2+), Fe(2+), Fe(3+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), Pb(2+), NO(3-), SO4(2-), PO4(3-), IO3(-), ClO(-) and Cl(-) by a 6-fold fluorescence enhancement. The working concentration of CrO4(2-) was from 0 to 90 ?M with a detection limit of 3.6??10(-7) mol/L. The detection could be carried out in water matrix and within a wide pH range (approximately from 2 to 12), and a large number of environmentally and biologically relevant ions including Cr(3+) showed no significant interferences with the detection. The sensing mechanism was explored by reversibility and LC/MS analysis, as well as Job's plot experiment, and the results suggested that the recognition was based on the oxidation of the primary alcohol in the structure of the sensor by the Cr(VI) sources. PMID:25686770

  5. A simple PCR-based fluorometric system for detection of mutant fusion DNAs using a quencher-free fluorescent DNA probe and graphene oxide.

    PubMed

    Roh, Kyoungmin; Kim, Dong-Min; Lee, Eun Hee; Kim, Hyoseon; Park, Hyung Soon; Jang, Ja-Hyun; Hwang, Sang-Hyun; Kim, Dong-Eun

    2015-04-25

    We propose a facile fluorometric system for detection of gene mutations using graphene oxide (GO). A fluorescent probe DNA anneals to a specific mutant gene and is degraded by the 5'? 3' exonuclease activity of Taq polymerase during PCR, and the released fluorophore retains fluorescence after addition of GO without quenching. PMID:25797656

  6. A simple and sensitive fluorescence based biosensor for the determination of uric acid using H2O2-sensitive quantum dots/dual enzymes.

    PubMed

    Azmi, Nur Ellina; Ramli, Noor Izaanin; Abdullah, Jaafar; Abdul Hamid, Mohammad Azmi; Sidek, Hamidah; Abd Rahman, Samsulida; Ariffin, Nurhayati; Yusof, Nor Azah

    2015-05-15

    A novel optical detection system consisting of combination of uricase/HRP-CdS quantum dots (QDs) for the determination of uric acid in urine sample is described. The QDs was used as an indicator to reveal fluorescence property of the system resulting from enzymatic reaction of uricase and HRP (horseradish peroxidase), which is involved in oxidizing uric acid to allaintoin and hydrogen peroxide. The hydrogen peroxide produced was able to quench the QDs fluorescence, which was proportional to uric acid concentration. The system demonstrated sufficient activity of uricase and HRP at a ratio of 5U:5U and pH 7.0. The linearity of the system toward uric acid was in the concentration range of 125-1000 M with detection limit of 125 M. PMID:25113659

  7. Fluorescence-Based Sensors

    NASA Astrophysics Data System (ADS)

    Orellana, Guillermo

    The natural luminescent phenomena (from the Latin words "lumen" and "essentia", i.e., "made of light") such as northern lights (aurora borealis), marine brightness, glow-worms, shining putrid fish scales, "bluish"- appearing water when contained in certain wooden cups (quinine fluorescence), some stones heated at high temperatures with reducing agents (BaS phosphorescence), or light emitted while crushing sugar (triboluminescence) already fascinated our ancestors. Nowadays we understand that ultraviolet and visible emission of light originates from a competitive deactivation pathway of the lowest electronic excited state of atoms and molecules that produces the so called luminescence (the sub-terms fluorescence and phosphorescence just designate whether the return of the excited to the ground state is an "allowed" or "forbidden" process, namely it is fast or slow, the loosely-defined border between them being a 1-?s-1 rate constant). Actually, luminescence is the only method to generate light in the known Universe regardless it is powered by the nuclear reactions in the stars, the ohmical heating in bulbs, an electric discharge, the absorption of light or a (bio)chemical reaction (chemiluminescence).

  8. Evaluating Sense Codon Reassignment with a Simple Fluorescence Screen.

    PubMed

    Biddle, Wil; Schmitt, Margaret A; Fisk, John D

    2015-12-22

    Understanding the interactions that drive the fidelity of the genetic code and the limits to which modifications can be made without breaking the translational system has practical implications for understanding the molecular mechanisms of evolution as well as expanding the set of encodable amino acids, particularly those with chemistries not provided by Nature. Because 61 sense codons encode 20 amino acids, reassigning the meaning of sense codons provides an avenue for biosynthetic modification of proteins, furthering both fundamental and applied biochemical research. We developed a simple screen that exploits the absolute requirement for fluorescence of an active site tyrosine in green fluorescent protein (GFP) to probe the pliability of the degeneracy of the genetic code. Our screen monitors the restoration of the fluorophore of GFP by incorporation of a tyrosine in response to a sense codon typically assigned another meaning in the genetic code. We evaluated sense codon reassignment at four of the 21 sense codons read through wobble interactions in Escherichia coli using the Methanocaldococcus jannaschii orthogonal tRNA/aminoacyl tRNA synthetase pair originally developed and commonly used for amber stop codon suppression. By changing only the anticodon of the orthogonal tRNA, we achieved sense codon reassignment efficiencies between 1% (Phe UUU) and 6% (Lys AAG). Each of the orthogonal tRNAs preferentially decoded the codon traditionally read via a wobble interaction in E. coli with the exception of the orthogonal tRNA with an AUG anticodon, which incorporated tyrosine in response to both the His CAU and His CAC codons with approximately equal frequencies. We applied our screen in a high-throughput manner to evaluate a 10(9)-member combined tRNA/aminoacyl tRNA synthetase library to identify improved sense codon reassigning variants for the Lys AAG codon. A single rapid screen with the ability to broadly evaluate reassignable codons will facilitate identification and improvement of the combinations of sense codons and orthogonal pairs that display efficient reassignment. PMID:26536053

  9. Binding strength of porphyrin-gold nanoparticle hybrids based on number and type of linker moieties and a simple method to calculate inner filter effects of gold nanoparticles using fluorescence spectroscopy.

    PubMed

    Shaikh, Ahson J; Rabbani, Faiz; Sherazi, Tauqir A; Iqbal, Zafar; Mir, Sadullah; Shahzad, Sohail A

    2015-02-19

    Gold nanoparticle-porphyrin assemblies were formed by binding functionalized porphyrins to gold nanoparticles (Au-NPs). Spectroscopic properties of hybrids and binding strength of porphyrins to Au-NPs were observed based on number and type of linker moieties using fluorescence spectroscopy. Binding appears to be dependent on number rather than type of linker moieties present on the porphyrin molecules, as tetraaminophenyl porphyrin shows the highest binding among the molecules we studied and causes agglomeration of nanoparticles due to presence of four linker groups. The inner filter effects of Au-NPs are considerably high due to their high extinction coefficient and cause large errors in the evaluation of quenching efficiencies. We have described a very simple method to calculate the inner filter effects of Au-NPs by first loading them with porphyrins and then replacing them with nonfluorescent ligands. The difference in the fluorescence of unbound porphyrins in the presence and absence of Au-NPs describes their inner filter effects. PMID:25611751

  10. Simple and rapid determination of the enzyme kinetics of HIV-1 reverse transcriptase and anti-HIV-1 agents by a fluorescence based method.

    PubMed

    Silprasit, Kun; Thammaporn, Ratsupa; Tecchasakul, Supanna; Hannongbua, Supa; Choowongkomon, Kiattawee

    2011-02-01

    Highly sensitive fluorescence dye, PicoGreen, can detect a small amount of polynucleotide duplex. The DNA/RNA duplex is a reaction product of HIV-1 reverse transcriptase (HIV-RT), which demonstrated that the HIV-1 RT kinetics and anti-HIV agent efficiency could be determined rapidly by a fluorometric method using PicoGreen dye. A small amount of dye is enough to test a minimal quantity of substrate to rapidly determine the enzyme kinetic properties K(m) and V(max). This method was applied to screen HIV-RT inhibitors. The enzymatic inhibition of 11 dipyridodiazepinone derivative inhibitors was compared. The IC(50) of all compounds was determined and it was found that two showed high inhibition efficiency. In addition, this method is applied for screening of many natural agents belonging to diverse sources. The proposed fluorometric method using PicoGreen dye is an easy, rapid, and sensitive detection method to determine HIV-RT activity, monitor enzyme kinetics and perform high-throughput screening of anti-HIV-1 RT agents. PMID:21167206

  11. Highly sensitive and simple fluorescence staining of proteins in sodium dodecyl sulfate-polyacrylamide-based gels by using hydrophobic tail-mediated enhancement of fluorescein luminescence.

    PubMed

    Kang, Chulhun; Kim, Hyun Jung; Kang, Donghoon; Jung, Duk Young; Suh, Myungkoo

    2003-10-01

    Fluorescein has an extremely low luminescence intensity in acidic aqueous media. However, when it was bound to proteins, subsequent increase of luminescence intensity took place. Furthermore, when a hydrophobic tail, such as aliphatic hydrocarbons, was introduced to fluorescein, more dramatic increase of luminescence intensity was observed upon binding to proteins. In the present study, by utilizing this luminescence enhancement, three hydrophobic fluorescein dyes (5-dodecanoyl amino fluorescein, 5-hexadecanoyl amino fluorescein, and 5-octadecanoyl amino fluorescein) were examined as noncovalent fluorescent stains of protein bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Effective incorporation of the dyes to proteins in gels was accomplished either simply by adding dyes at the protein fixation step, or by treating gels with a staining solution after the fixation. The sensitivity of this staining method using the fluorescein derivatives was approximately 1 ng/band for most proteins. For some cases, protein bands containing as low as 0.1 ng were successfully visualized. In addition, the detection sensitivity showed much less protein-to-protein variation than silver staining. This new staining method was also successfully applied to two-dimensional electrophoresis of rat brain proteins. Its overall sensitivity was comparable to that of silver staining. PMID:14595675

  12. Polyelectrolyte-Based Fluorescent Sensors

    NASA Astrophysics Data System (ADS)

    Ogawa, K.; Achyuthan, K. E.; Chemburu, S.; Ji, E.; Liu, Y.; Lopez, G. P.; Schanze, S.; Whitten, D. G.

    Chemo- and biosensors based on fluorescent polyelectrolytes are discussed. The amplified quenching of conjugated polyelectrolytes by oppositely charged quenchers can be used in bioassays to monitor activities of protease, phospholipase A2, and phospholipase C by varying substrates. Enhanced fluorescence yields from cyanine dye J aggregates formed by supramolecular self-assembly between the dyes and polysaccharides can be utilized in assays to monitor glycosidase activity. In addition solution based polyelectrolyte sensors, amplified quenching properties of surface grafted conjugated polyelectrolytes by energy transfer and electron transfer type quenchers are compared.

  13. A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.

    PubMed

    Zeller, Perrine; Ploux, Olivier; Mjean, Annick

    2016-03-01

    Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria. PMID:26780689

  14. Fluorescent sensors based on boronic acids

    NASA Astrophysics Data System (ADS)

    Cooper, Christopher R.; James, Tony D.

    1999-05-01

    Sensor systems have long been needed for detecting the presence in solution of certain chemically or biologically important species. Sensors are used in a wide range of applications from simple litmus paper that shows a single color change in acidic or basic environments to complex biological assays that use enzymes, antibodies and antigens to display binding events. With this work the use of boronic acids in the design and synthesis of sensors for saccharides (diols) will be presented. The fluorescent sensory systems rely on photoinduced electron transfer (PET) to modulate the observed fluorescence. When saccharides form cyclic boronate esters with boronic acids, the Lewis acidity of the boronic acid is enhanced and therefore the Lewis acid-base interaction between the boronic acid and a neighboring amine is strengthened. The strength of this acid-base interaction modulates the PET from the amine (acting as a quencher) to anthracene (acting as a fluorophore). These compounds show increased fluorescence at neutral pH through suppression of the PET from nitrogen to anthracene on saccharide binding. The general strategy for the development of saccharide selective systems will be discussed. The potential of the boronic acid based systems will be illustrated using the development of glucose and glucosamine selective fluorescent sensors as examples.

  15. Lens-based fluorescence nanoscopy.

    PubMed

    Eggeling, Christian; Willig, Katrin I; Sahl, Steffen J; Hell, Stefan W

    2015-05-01

    The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (~200nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous developments for studying the living cell, as the move from microscopy to super-resolution microscopy or 'nanoscopy' offers opportunities to study problems in biophysical and biomedical research at a new level of detail. This review describes the principles and modalities of present fluorescence nanoscopes, as well as their potential for biophysical and cellular experiments. All the existing nanoscopy variants separate neighboring features by transiently preparing their fluorescent molecules in states of different emission characteristics in order to make the features discernible. Usually these are fluorescent 'on' and 'off' states causing the adjacent molecules to emit sequentially in time. Each of the variants can in principle reach molecular spatial resolution and has its own advantages and disadvantages. Some require specific transitions and states that can be found only in certain fluorophore subfamilies, such as photoswitchable fluorophores, while other variants can be realized with standard fluorescent labels. Similar to conventional far-field microscopy, nanoscopy can be utilized for dynamical, multi-color and three-dimensional imaging of fixed and live cells, tissues or organisms. Lens-based fluorescence nanoscopy is poised for a high impact on future developments in the life sciences, with the potential to help solve long-standing quests in different areas of scientific research. PMID:25998828

  16. A simple procedure to improve the surface passivation for single molecule fluorescence studies

    NASA Astrophysics Data System (ADS)

    Pan, Hai; Xia, Yifan; Qin, Meng; Cao, Yi; Wang, Wei

    2015-07-01

    The single-molecule fluorescence technique is becoming a general and mature tool to probe interactions and dynamics of biomolecules with ultra high precision and accuracy. However, nonspecific adsorption of biomolecules to the flow cells remains a major experimental riddle for the study of many complex biological systems, especially those exhibiting low binding affinity and presenting with weakly populated intermediates. Many novel surface passivation methods have been introduced to reduce nonspecific interactions. Here, we present an effective and inexpensive method to significantly reduce nonspecific binding of biomolecules in conventional poly (ethylene glycol) (PEG)-based surface passivation protocols, without additional exogenous effects. In particular, we propose a simple 10 min Tween-20 treatment for the PEG passivated surface, which could further increase the hydrophilicity of the surface and thus promote passivation efficacy by about 5 to 10 times. We anticipate that this new procedure will find broad practical applications and extend the current reaches of single-molecule fluorescence studies.

  17. Selection of fluorescent DNA dyes for real-time LAMP with portable and simple optics.

    PubMed

    Seyrig, Gregoire; Stedtfeld, Robert D; Tourlousse, Dieter M; Ahmad, Farhan; Towery, Keara; Cupples, Alison M; Tiedje, James M; Hashsham, Syed A

    2015-12-01

    Loop-mediated isothermal amplification (LAMP) is increasingly used for point-of-care nucleic acid based diagnostics. LAMP can be monitored in real-time by measuring the increase in fluorescence of DNA binding dyes. However, there is little information comparing the effect of various fluorescent dyes on signal to noise ratio (SNR) or threshold time (Tt). This information is critical for implementation with field deployable diagnostic tools that require small, low power consumption, robust, and inexpensive optical components with reagent saving low volume reactions. In this study, SNR and Tt during real-time LAMP was evaluated with eleven fluorescent dyes. Of all dyes tested, SYTO-82, SYTO-84, and SYTOX Orange resulted in the shortest Tt, and SYTO-81 had the widest range of working concentrations. The optimized protocol detected 10 genome copies of Mycobacterium tuberculosis in less than 10min, 10 copies of Giardia intestinalis in ~20min, and 10 copies of Staphylococcus aureus or Salmonella enterica in less than 15min. Results demonstrate that reaction efficiency depends on both dye type and concentration and the selected polymerase. The optimized protocol was evaluated in the Gene-Z device, a hand-held battery operated platform characterized via simple and low cost optics, and a multiple assay microfluidic chip with micron volume reaction wells. Compared to the more conventional intercalating dye (SYBR Green), reliable amplification was only observed in the Gene-Z when using higher concentrations of SYTO-81. PMID:26554941

  18. Ultrasound-modulated fluorescence based on fluorescent microbubbles

    NASA Astrophysics Data System (ADS)

    Liu, Yuan; Feshitan, Jameel A.; Wei, Ming-Yuan; Borden, Mark A.; Yuan, Baohong

    2014-08-01

    Ultrasound-modulated fluorescence (UMF) imaging has been proposed to provide fluorescent contrast while maintaining ultrasound resolution in an optical-scattering medium (such as biological tissue). The major challenge is to extract the weakly modulated fluorescent signal from a bright and unmodulated background. UMF was experimentally demonstrated based on fluorophore-labeled microbubble contrast agents. These contrast agents were produced by conjugating N-hydroxysuccinimide (NHS)-ester-attached fluorophores on the surface of amine-functionalized microbubbles. The fluorophore surface concentration was controlled so that a significant self-quenching effect occurred when no ultrasound was applied. The intensity of the fluorescent emission was modulated when microbubbles were oscillated by ultrasound pulses, presented as UMF signal. Our results demonstrated that the UMF signals were highly dependent on the microbubbles' oscillation amplitude and the initial surface fluorophore-quenching status. A maximum of 42% UMF modulation depth was achieved with a single microbubble under an ultrasound peak-to-peak pressure of 675 kPa. Further, UMF was detected from a 500-?m tube filled with contrast agents in water and scattering media with ultrasound resolution. These results indicate that ultrasound-modulated fluorescent microbubble contrast agents can potentially be used for fluorescence-based molecular imaging with ultrasound resolution in the future.

  19. Ultrasound-modulated fluorescence based on fluorescent microbubbles

    PubMed Central

    Liu, Yuan; Feshitan, Jameel A.; Wei, Ming-Yuan; Borden, Mark A.; Yuan, Baohong

    2014-01-01

    Abstract. Ultrasound-modulated fluorescence (UMF) imaging has been proposed to provide fluorescent contrast while maintaining ultrasound resolution in an optical-scattering medium (such as biological tissue). The major challenge is to extract the weakly modulated fluorescent signal from a bright and unmodulated background. UMF was experimentally demonstrated based on fluorophore-labeled microbubble contrast agents. These contrast agents were produced by conjugating N-hydroxysuccinimide (NHS)-ester-attached fluorophores on the surface of amine-functionalized microbubbles. The fluorophore surface concentration was controlled so that a significant self-quenching effect occurred when no ultrasound was applied. The intensity of the fluorescent emission was modulated when microbubbles were oscillated by ultrasound pulses, presented as UMF signal. Our results demonstrated that the UMF signals were highly dependent on the microbubbles oscillation amplitude and the initial surface fluorophore-quenching status. A maximum of ?42% UMF modulation depth was achieved with a single microbubble under an ultrasound peak-to-peak pressure of 675kPa. Further, UMF was detected from a 500-?m tube filled with contrast agents in water and scattering media with ultrasound resolution. These results indicate that ultrasound-modulated fluorescent microbubble contrast agents can potentially be used for fluorescence-based molecular imaging with ultrasound resolution in the future. PMID:25104407

  20. Fluorescent sensors based on bacterial fusion proteins

    NASA Astrophysics Data System (ADS)

    Prats Mateu, Batirtze; Kainz, Birgit; Pum, Dietmar; Sleytr, Uwe B.; Toca-Herrera, Jos L.

    2014-06-01

    Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins.

  1. Multiphoton excitation fluorescence correlation spectroscopy of fluorescent DNA base analogs

    NASA Astrophysics Data System (ADS)

    Katilius, Evaldas; Woodbury, Neal W.

    2004-06-01

    Two- and three-photon excitation was used to investigate the properties of two fluorescent DNA base analogs: 2-aminopurine and 6-methylisoxanthopterin. 2-aminopurine is a widely used fluorescent analog of the DNA base adenine. Three-photon excitation of 2-aminopurine is achievable by using intense femtosecond laser pulses in 850-950 nm spectral region. Interestingly, the three-photon excitation spectrum is blue-shifted relative to the three-times-wavelength single-photon excitation spectrum. The maximum of the absorbance band in the UV is at 305 nm, while the three-photon excitation spectrum has a maximum at around 880 nm. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2-aminopurine for DNA-protein interaction studies. However, due to relatively small three-photon absorption cross-section, a good signal-to-noise fluorescence correlation curves take very long time to obtain. Fluorescence properties of 6-methylisoxanthopterin, the fluorescent analog of guanine, were investigated using two-photon excitation. This molecule has the lowest energy absorption band centered around 350 nm, thus, two-photon excitation is attainable using 700 to 760 nm output of Ti-sapphire laser. The excitation spectrum of this molecule in the infrared well matches the doubled-wavelength single-photon excitation spectrum in the UV. The high fluorescence quantum yield of 6-methylisoxanthopterin allows efficient fluorescence correlation measurements and makes this molecule a very good candidate for using in in vitro DNA-protein interaction studies.

  2. A simple dental caries detection system using full spectrum of laser-induced fluorescence

    NASA Astrophysics Data System (ADS)

    Rocha-Cabral, Renata Maciel; Mendes, Fausto Medeiros; Maldonado, Edison Puig; Zezell, Denise Maria

    2015-06-01

    Objectives: to develop an apparatus for the detection of early caries lesions in enamel using the full extent of the tooth fluorescence spectrum, through the integration of a laser diode, fiber optics, filters and one portable spectrometer connected to a computer, all commercially available; to evaluate the developed device in clinical and laboratory tests, and compare its performance with commercial equipment. Methods: clinical examinations were performed in patients with indication for exodontics of premolars. After examinations, the patients underwent surgery and the teeth were stored individually. The optical measurements were repeated approximately two months after extraction, on the same sites previously examined, then histological analysis was carried out. Results: the spectral detector has presented high specificity and moderate sensitivity when applied to differentiate between healthy and damaged tissues, with no significant differences from the performance of the commercial equipment. The developed device is able to detect initial damages in enamel, with depth of approximately 300 μm. Conclusions: we successfully demonstrated the development of a simple and portable system based in laser-induced fluorescence for caries detection, assembled from common commercial parts. As the spectral detector acquires a complete recording of the spectrum from each tissue, it is possible to use it for monitoring developments of caries lesions.

  3. A simple and sensitive approach for ochratoxin A detection using a label-free fluorescent aptasensor.

    PubMed

    Lv, Zhenzhen; Chen, Ailiang; Liu, Jinchuan; Guan, Zheng; Zhou, Yu; Xu, Siyuan; Yang, Shuming; Li, Cheng

    2014-01-01

    Ochratoxin A(OTA) is found to be one of the predominant contaminating mycotoxins in a wide variety of food commodities. To avoid the risk of OTA consumption, the detection and quantitation of OTA level are of great significance. Based on the fact that ssDNA aptamer has the ability to form a double-strand structure with its complementary sequence, a simple and rapid aptamer-based label-free approach for highly sensitive and selective fluorescence detection of OTA was developed by using ultra-sensitive double-strand DNA specific dyes PicoGreen. The results showed that as low as 1 ng/mL of OTA could be detected with a dynamic range of more than 5 orders of magnitude which satisfies the requirements for OTA maximum residue limit in various food regulated by European Commission. With the specificity of aptamer, the assay exhibited high selectivity for OTA against two other analogues (N-acetyl-l-phenylalanine and zearalenone). We also tested the aptasensor practicability using real sample of 1% beer spiked with a series of concentration of OTA and the results show good tolerance to matrix effect. All detections could be achieved in less than 30 min, which provides a simple, quick and sensitive detection method for OTA screening in food safety and could be easily extend to other small molecular chemical compounds detection which aptamer has been selected. PMID:24465818

  4. A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates.

    PubMed

    Koshi, Yoichiro; Nakata, Eiji; Yamane, Hiroki; Hamachi, Itaru

    2006-08-16

    Because sugar and its derivatives play important roles in various biological phenomena, the rapid and high-throughput analysis of various glycoconjugates is keenly desirable. We describe herein the construction of a novel fluorescent lectin array for saccharide detection using a supramolecular hydrogel matrix. In this array, the fluorescent lectins were noncovalently fixed under semi-wet conditions to suppress the protein denaturation. It is demonstrated by fluorescence titration and fluorescence lifetime experiments that the immobilized lectins act as a molecular recognition scaffold in the hydrogel matrix, similar to that in aqueous solution. That is, a bimolecular fluorescence quenching and recovery (BFQR) method can successfully operate under both conditions. This enables one to fluorescently read-out a series of saccharides on the basis of the recognition selectivity and affinity of the immobilized lectins without tedious washing processes and without labeling the target saccharides. Simple and high-throughput sensing and profiling were carried out using the present lectin array for diverse glycoconjugates, which not only included a simple glucose, but also oligosaccharides, and glycoproteins, and, furthermore, the pattern recognition and profiling of several types of cell lysates were also accomplished. PMID:16895406

  5. A simple and compact fluorescence detection system for capillary electrophoresis and its application to food analysis.

    PubMed

    Zhai, Haiyun; Yuan, Kaisong; Yu, Xiao; Chen, Zuanguang; Liu, Zhenping; Su, Zihao

    2015-10-01

    A novel fluorescence detection system for CE was described and evaluated. Two miniature laser pointers were used as the excitation source. A Y-style optical fiber was used to transmit the excitation light and a four-branch optical fiber was used to collect the fluorescence. The optical fiber and optical filter were imported into a photomultiplier tube without any extra fixing device. A simplified PDMS detection cell was designed with guide channels through which the optical fibers were easily aligned to the detection window of separation capillary. According to different requirements, laser pointers and different filters were selected by simple switching and replacement. The fluorescence from four different directions was collected at the same detecting point. Thus, the sensitivity was enhanced without peak broadening. The fluorescence detection system was simple, compact, low-cost, and highly sensitive, with its functionality demonstrated by the separation and determination of red dyes and fluorescent whitening agents. The detection limit of rhodamine 6G was 7.7 nM (S/N = 3). The system was further applied to determine illegal food dyes. The CE system is potentially eligible for food safety analysis. PMID:26109527

  6. A fluorescence high-temperature sensor based on fluorescence lifetime

    NASA Astrophysics Data System (ADS)

    Wu, Jinling; Wang, Yutian; Wang, Xinian

    2006-11-01

    A kind of fluorescence optic-fiber temperature sensor is devised based on the alexandrite crystal. In this system, a new optic- fiber probe fabrication techniques is proposed. This system is particularly adapted to the temperature measurement in the range of room temperature to 650°C. During the cause of experimentation, using the PLD-PMTR (termed the Pulse Modulated Phase-locked detection with Two References) signal processing scheme. This temperature measurement method is proved to be effective and useful for its highly resolution and precision. It ensured the detected fluorescence signal to noise ratio was high enough to be measurable when the temperature is raised to 650°C.

  7. Multiphoton excitation of fluorescent DNA base analogs.

    PubMed

    Katilius, Evaldas; Woodbury, Neal W

    2006-01-01

    Multiphoton excitation was used to investigate properties of the fluorescent DNA base analogs, 2-aminopurine (2AP) and 6-methylisoxanthopterin (6MI). 2-aminopurine, a fluorescent analog of adenine, was excited by three-photon absorption. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2AP for DNA-protein interaction studies. However, high excitation power and long integration times needed to acquire high signal-to-noise fluorescence correlation curves render three-photon excitation FCS of 2AP not very useful for studying DNA base dynamics. The fluorescence properties of 6-methylisoxanthopterin, a guanine analog, were investigated using two-photon excitation. The two-photon absorption cross-section of 6MI was estimated to be about 2.5 x 10(-50) cm(4)s (2.5 GM units) at 700 nm. The two-photon excitation spectrum was measured in the spectral region from 700 to 780 nm; in this region the shape of the two-photon excitation spectrum is very similar to the shape of single-photon excitation spectrum in the near-UV spectral region. Two-photon excitation of 6MI is suitable for fluorescence correlation measurements. Such measurements can be used to study DNA base dynamics and DNA-protein interactions over a broad range of time scales. PMID:16965161

  8. Multiphoton excitation of fluorescent DNA base analogs

    NASA Astrophysics Data System (ADS)

    Katilius, Evaldas; Woodbury, Neal W.

    2006-07-01

    Multiphoton excitation was used to investigate properties of the fluorescent DNA base analogs, 2-aminopurine (2AP) and 6-methylisoxanthopterin (6MI). 2-aminopurine, a fluorescent analog of adenine, was excited by three-photon absorption. Fluorescence correlation measurements were attempted to evaluate the feasibility of using three-photon excitation of 2AP for DNA-protein interaction studies. However, high excitation power and long integration times needed to acquire high signal-to-noise fluorescence correlation curves render three-photon excitation FCS of 2AP not very useful for studying DNA base dynamics. The fluorescence properties of 6-methylisoxanthopterin, a guanine analog, were investigated using two-photon excitation. The two-photon absorption cross-section of 6MI was estimated to be about 2.510-50 cm4s (2.5 GM units) at 700 nm. The two-photon excitation spectrum was measured in the spectral region from 700 to 780 nm; in this region the shape of the two-photon excitation spectrum is very similar to the shape of single-photon excitation spectrum in the near-UV spectral region. Two-photon excitation of 6MI is suitable for fluorescence correlation measurements. Such measurements can be used to study DNA base dynamics and DNA-protein interactions over a broad range of time scales.

  9. Handheld Fluorescence Microscopy based Flow Analyzer.

    PubMed

    Saxena, Manish; Jayakumar, Nitin; Gorthi, Sai Siva

    2016-03-01

    Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second. PMID:26715517

  10. A simple and general route for monofunctionalization of fluorescent and magnetic nanoparticles using peptides.

    PubMed

    Clarke, Samuel; Tamang, Sudarsan; Reiss, Peter; Dahan, Maxime

    2011-04-29

    Nanoparticles are now utilized in many diverse biological and medical applications. Despite this, it remains challenging to tailor their surface for specific molecular targeting while maintaining high biocompatibility. To address this problem, we evaluate a phytochelatin-related peptide surface coating to produce functional and biocompatible nanoparticles (NPs) based on fluorescent InP/ZnS and CdSe/ZnS or superparamagnetic FePt and Fe(3)O(4). Using a combination of transmission electron microscopy, size-exclusion chromatography and gel electrophoresis (GE), we demonstrate the excellent colloidal properties of the peptide-coated NPs (pNPs) and the compact nature of the coating (?4nm thickness). We develop a simple protocol for the monofunctionalization of the pNPs with targeting biomolecules, by combining covalent conjugation with GE purification. We then employ functionalized InP/ZnS pNPs in a live-cell, single-molecule imaging application to specifically target and detect individual proteins in the cell membrane. These findings showcase the versatility of the peptides for preparing compact NPs of various compositions and sizes, which are easily functionalized, and suitable for a broad range of biomedical applications. PMID:21411925

  11. a Simple, Cost Effective Raman-Fluorescence Spectrometer for Use in Laboratory and Field Experiments

    NASA Astrophysics Data System (ADS)

    Marshall, Frank E.; Pride, Michael A.; Rojo, Michellle; Brinker, Katelyn R.; Walker, Zachary; Storrie-Lombardi, Michael; Mormile, Melanie R.; Grubbs, G. S., II

    2015-06-01

    Research, design, construction, and operation of a portable mixed Raman and Fluorescence type spectrometer implemented by the Missouri University of Science and Technology's Mars Rover Design Team will be presented. This spectrometer has been built for the team's annual competition. The spectrometer, completely built by undergraduates, is designed to use a 50 mW, 532 nm constant waveform laser to probe a sample of soil to find bacteria or bio-markers. However, initial tests of the spectrometer were carried out in a laboratory environment making the spectrometer also suitable for simple undergraduate physical chemistry or chemical physics laboratory experiments. The final cost of the device is roughly 2100, weighs 1.4 kg, and is 22.9 cm x 22.6 cm in size. Integrating the spectrometer with a computer database, results from the competition, complications of fitting mixed Raman-Fluorescence spectra, and future ideas/improvements will also be discussed.

  12. A fluorescent chemosensor based on naphthol for detection of Zn(2.).

    PubMed

    Lin, Hsiang-Yi; Chen, Ting-Yu; Liu, Chih-Kai; Wu, An-Tai

    2016-02-01

    A simple naphthol-based fluorescent receptor 1 was prepared and evaluated for its fluorescence response to heavy metal ions. Receptor 1 exhibits an 'off-on-type' mode with high selectivity in the presence of Zn(2+) ion. The selectivity of 1 for Zn(2+) is the consequence of combined effects of chelation-enhanced fluorescence (CHEF), C?=?N isomerization and ?-? stacking interaction between the two naphthalene rings. Copyright 2015 John Wiley & Sons, Ltd. PMID:26032465

  13. Implantable fluorescence-based glucose sensor development

    NASA Astrophysics Data System (ADS)

    Ibey, Bennett L.; Yadavalli, Vamsi K.; Thomas, Hope R.; Rounds, Rebecca M.; Pishko, Michael V.; Cote, Gerard L.

    2005-03-01

    An implantable sensor is being created that allows measurement of blood glucose through fluorescent detection of an embedded chemical assay. The sensor is based on the competitive binding reaction between the protein Concanavalin A and various saccharide molecules, specifically a glycodendrimer and glucose. Previous studies have shown the ability of an embedded chemical assay using Con A and dextran with shorter wavelength dyes to both sense changes in glucose and generate sufficient fluorescent emission to pass through the dermal tissue. However, due to the chemical constituents of the assay, multivalent binding was evident resulting in poor spectral change due to glucose within the biological range. Use of a glycodendrimer and longer wavelength dyes has improved the sensor"s spectral change due to glucose and the overall signal to noise ratio of the sensor. In this work, a description of this sensor and the results obtained from it will be presented showing a large dynamic range of fluorescence with glucose.

  14. Ultrasensitive turn-on fluorescent detection of trace thiocyanate based on fluorescence resonance energy transfer.

    PubMed

    Song, Juan; Wu, Fang-Ying; Wan, Yi-Qun; Ma, Li-Hua

    2015-01-01

    Thiocyanate (SCN(-)) is a small anion byproduct of cyanide metabolism. Several methods have been reported to measure SCN(-) above the micromolar level. However, SCN(-) is derived from many sources such as cigarettes, waste water, food and even car exhaust and its effect is cumulative, which makes it necessary to develop methods for the detection of trace SCN(-). In this paper, a simple and ultrasensitive turn-on fluorescence assay of trace SCN(-) is established based on the fluorescence resonance energy transfer (FRET) between gold nanoparticles (AuNPs) and fluorescein. The detection limit is 0.09 nM, to the best of our knowledge, which has been the lowest detection LOD ever without the aid of costly instrumentation. The fluorescence of fluorescein is significantly quenched when it is attached to the surface of AuNPs. Upon the addition of SCN(-), the fluorescence is turned on due to the competition action between SCN(-) and fluorescein towards the surface of AuNPs. Under an optimum pH, AuNPs size and concentration, incubation time, the fluorescence enhancement efficiency [(IF-I0)/I0] displays a linear relationship with the concentration of SCN(-) in the range of 1.0 nM to 40.0 nM. The fluorescein-AuNP sensor shows absolutely high selectivity toward SCN(-) than other 16 anions. The common metal ions, amino acids and sugars have no obvious interference effects. The accuracy and precision were evaluated based on the recovery experiments. The cost effective sensing system is successfully applied for the determination of SCN(-) in milk products and saliva samples. PMID:25476353

  15. Optical oxygen sensor based on time-resolved fluorescence

    NASA Astrophysics Data System (ADS)

    Chu, Cheng-Shane; Chu, Ssu-Wei

    2015-07-01

    A new, simple signal processing, low-cost technique for the fabrication of a portable oxygen sensor based on time-resolved fluorescence is described. The sensing film uses the oxygen sensing dye platinum meso-tetra (pentfluorophenyl) porphyrin (PtTFPP) embedded in a polymer matrix. The experimental results reveal that the PtTFPP-doped oxygen sensor has a sensitivity of 2.2 in the 0-100% range. A preparation procedure for coating the photodiodes with the oxygen sensor film that produces repetitive and reliable sensing devices is proposed. The developed time-resolved optical oxygen sensor is portable, low-cost, has simple signal processing, and lacks optical filter elements. It is a cost-effective alternative to traditional electrochemical-based oxygen sensors and provides a platform for other optical based sensors.

  16. A simple and sensitive HPLC fluorescence method for determination of tadalafil in mouse plasma

    PubMed Central

    Farthing, Christine A; Farthing, Don E; Koka, Saisudha; Larus, Terri; Fakhry, Itaf; Xi, Lei; Kukreja, Rakesh C; Sica, Domenic; Gehr, Todd WB

    2010-01-01

    A simple and sensitive high-performance liquid chromatographic (HPLC) method utilizing fluorescence detection was developed for the determination of the phosphodiesterase type 5 inhibitor tadalafil in mouse plasma. This method utilizes a simple sample preparation (protein precipitation) with high recovery of tadalafil (?98%), which eliminates the need for an internal standard. For constituent separation, the method utilized a monolithic C18 column and a flow rate of 1.0 ml/min with a mobile phase gradient consisting of aqueous trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and acetonitrile. The method calibration was linear for tadalafil in mouse plasma from 100 to 2000 ng/mL (r > 0.999) with a detection limit of approximately 40 ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275 nm with monitoring of the emission wavelength at 335 nm. The intra- and inter-day precision (relative standard deviation, RSD) values for tadalafil in mouse plasma were less than 14%, and the accuracy (percent error) was within -14% of the nominal concentration. The method was utilized on mouse plasma samples from research evaluating the potential cardioprotective effects of tadalafil on mouse heart tissue exposed to doxorubicin, a chemotherapeutic drug with reported cardiotoxic effects. PMID:20801090

  17. A simple and highly sensitive fluorescence assay for microRNAs.

    PubMed

    Shen, Wei; Yeo, Kiat Huei; Gao, Zhiqiang

    2015-03-21

    Herein, we have reported a simple and highly sensitive fluorescence assay for the detection of microRNAs (miRNAs). The assay uses a duplex-specific nuclease (DSN) to amplify the fluorescence signal and magnetic beads (MBs) to completely remove the unreacted DNA detection probes. Briefly, fluorescein-capped DNA detection probes were first conjugated to the MBs. The use of the MBs produced a very low background signal since all the unreacted DNA probes can be conveniently removed from the solution by using a permanent magnet. During the assaying process, target miRNA strands hybridized with the DNA capture probes to form miRNA-DNA heteroduplexes. The DSN then selectively cleaved the DNA probes in the miRNA-DNA duplexes and release the target miRNA strands back into the solution, thereby establishing a target recycling amplification mechanism - a cumulative signal amplification process. A much-amplified fluorescence signal was obtained in the presence of traces of the target miRNA. In addition, a negligible background signal was conveniently attained by the complete removal of the unreacted DNA detection probes so that minute change in the fluorescence signal can be unambiguously detected. The negligible background signal in association with the accumulative signal amplification significantly lowered the detection limit and broadened the dynamic range of the assay. Moreover, the high specificity of the DSN to perfectly matched duplexes endowed this assay with good selectivity when analyzing target miRNAs with high sequence similarities. Successful attempts were made in applying the proposed assay to detect let-7a in total RNA extracted from cultured cells. PMID:25655238

  18. Simple, Script-Based Science Processing Archive

    NASA Technical Reports Server (NTRS)

    Lynnes, Christopher; Hegde, Mahabaleshwara; Barth, C. Wrandle

    2007-01-01

    The Simple, Scalable, Script-based Science Processing (S4P) Archive (S4PA) is a disk-based archival system for remote sensing data. It is based on the data-driven framework of S4P and is used for data transfer, data preprocessing, metadata generation, data archive, and data distribution. New data are automatically detected by the system. S4P provides services such as data access control, data subscription, metadata publication, data replication, and data recovery. It comprises scripts that control the data flow. The system detects the availability of data on an FTP (file transfer protocol) server, initiates data transfer, preprocesses data if necessary, and archives it on readily available disk drives with FTP and HTTP (Hypertext Transfer Protocol) access, allowing instantaneous data access. There are options for plug-ins for data preprocessing before storage. Publication of metadata to external applications such as the Earth Observing System Clearinghouse (ECHO) is also supported. S4PA includes a graphical user interface for monitoring the system operation and a tool for deploying the system. To ensure reliability, S4P continuously checks stored data for integrity, Further reliability is provided by tape backups of disks made once a disk partition is full and closed. The system is designed for low maintenance, requiring minimal operator oversight.

  19. Simple interface of high-performance liquid chromatography-atomic fluorescence spectrometry hyphenated system for speciation of mercury based on photo-induced chemical vapour generation with formic acid in mobile phase as reaction reagent.

    PubMed

    Yin, Yongguang; Liu, Jingfu; He, Bin; Shi, Jianbo; Jiang, Guibin

    2008-02-15

    Photo-induced chemical vapour generation (CVG) with formic acid in mobile phase as reaction reagent was developed as interface to on-line couple HPLC with atomic fluorescence spectrometry for the separation and determination of inorganic mercury, methylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg). In the developed procedure, formic acid in mobile phase was used to decompose organomercuries and reduce Hg(2+) to mercury cold vapour under UV irradiation. Therefore, no post-column reagent was used and the flow injection system in traditional procedure is omitted. A number of operating parameters including pH of mobile phase, concentration of formate, flow rate of mobile phase, length of PTFE reaction coil, flow rate of carrier gas and Na(2)S(2)O(3) in sample matrix were optimized. The limits of detection at the optimized conditions were 0.085, 0.033, 0.029 and 0.038 microg L(-1) for inorganic mercury, MeHg, EtHg and PhHg, respectively. The developed method was validated by determination of certified reference material DORM-2 and was further applied in analyses of seafood samples from Yantai port, China. The UV-CVG with formic acid simplifies the instrumentation and reduces the analytical cost significantly. PMID:18184614

  20. Quick and simple estimation of bacteria using a fluorescent paracetamol dimer-Au nanoparticle composite

    NASA Astrophysics Data System (ADS)

    Sahoo, Amaresh Kumar; Sharma, Shilpa; Chattopadhyay, Arun; Ghosh, Siddhartha Sankar

    2012-02-01

    Rapid, simple and sensitive detection of bacterial contamination is critical for safeguarding public health and the environment. Herein, we report an easy method of detection as well as enumeration of the bacterial cell number on the basis of fluorescence quenching of a non-antibacterial fluorescent nanocomposite, consisting of paracetamol dimer (PD) and Au nanoparticles (NPs), in the presence of bacteria. The composite was synthesized by reaction of paracetamol (p-hydroxyacetanilide) with HAuCl4. The Au NPs of the composite were characterized using UV-Vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and selected area electron diffraction analysis. The paracetamol dimer in the composite showed emission peak at 435 nm when excited at 320 nm. The method successfully detected six bacterial strains with a sensitivity of 100 CFU mL-1. The Gram-positive and Gram-negative bacteria quenched the fluorescence of the composite differently, making it possible to distinguish between the two. The TEM analysis showed interaction of the composite with bacteria without any apparent damage to the bacteria. The chi-square test established the accuracy of the method. Quick, non-specific and highly sensitive detection of bacteria over a broad range of logarithmic dilutions within a short span of time demonstrates the potential of this method as an alternative to conventional methods.Rapid, simple and sensitive detection of bacterial contamination is critical for safeguarding public health and the environment. Herein, we report an easy method of detection as well as enumeration of the bacterial cell number on the basis of fluorescence quenching of a non-antibacterial fluorescent nanocomposite, consisting of paracetamol dimer (PD) and Au nanoparticles (NPs), in the presence of bacteria. The composite was synthesized by reaction of paracetamol (p-hydroxyacetanilide) with HAuCl4. The Au NPs of the composite were characterized using UV-Vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and selected area electron diffraction analysis. The paracetamol dimer in the composite showed emission peak at 435 nm when excited at 320 nm. The method successfully detected six bacterial strains with a sensitivity of 100 CFU mL-1. The Gram-positive and Gram-negative bacteria quenched the fluorescence of the composite differently, making it possible to distinguish between the two. The TEM analysis showed interaction of the composite with bacteria without any apparent damage to the bacteria. The chi-square test established the accuracy of the method. Quick, non-specific and highly sensitive detection of bacteria over a broad range of logarithmic dilutions within a short span of time demonstrates the potential of this method as an alternative to conventional methods. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr11837h

  1. A simple and rapid protocol for measuring neutral lipids in algal cells using fluorescence.

    PubMed

    Storms, Zachary J; Cameron, Elliot; de la Hoz Siegler, Hector; McCaffrey, William C

    2014-01-01

    Algae are considered excellent candidates for renewable fuel sources due to their natural lipid storage capabilities. Robust monitoring of algal fermentation processes and screening for new oil-rich strains requires a fast and reliable protocol for determination of intracellular lipid content. Current practices rely largely on gravimetric methods to determine oil content, techniques developed decades ago that are time consuming and require large sample volumes. In this paper, Nile Red, a fluorescent dye that has been used to identify the presence of lipid bodies in numerous types of organisms, is incorporated into a simple, fast, and reliable protocol for measuring the neutral lipid content of Auxenochlorella protothecoides, a green alga. The method uses ethanol, a relatively mild solvent, to permeabilize the cell membrane before staining and a 96 well micro-plate to increase sample capacity during fluorescence intensity measurements. It has been designed with the specific application of monitoring bioprocess performance. Previously dried samples or live samples from a growing culture can be used in the assay. PMID:24961928

  2. Economic and simple system to combine single-spot photolysis and whole-field fluorescence imaging

    NASA Astrophysics Data System (ADS)

    Jaafari, Nadia; Henson, Mark; Graham, Jeremy; Canepari, Marco

    2013-06-01

    In recent years, the use of light emitting diodes (LEDs) has become commonplace in fluorescence microscopy. LEDs are economical and easy to couple to commercial microscopes, and they provide powerful and stable light that can be triggered by transistor-transistor logic pulses in the range of tens of microseconds or shorter. LEDs are usually installed on the epifluorescence port of the microscope to obtain whole-field illumination, which is ideal for fluorescence imaging. In contrast, photolysis or channelrhodopsin stimulation often requires localized illumination, typically achieved using lasers. Here we show that insertion of a long-pass (>411 nm) filter with an appropriately sized pinhole in the epifluorescence pathway, combined with dual UV/visible illumination, can produce efficient whole-field visible illumination and spot UV illumination of 15 to 20 μm. We tested our system by performing calcium imaging experiments combined with L-glutamate or N-methyl-D-aspartic acid (NMDA) photorelease in hippocampal neurons from brain slices or dissociated cultures, demonstrating the ability to obtain local activation of NMDA receptors exclusively in the illuminated spot. The very inexpensive and simple system that we report here will allow many laboratories with limited budgets to run similar experiments in a variety of physiological applications.

  3. A Fluorescence Based Dissolved Oxygen Sensor

    NASA Astrophysics Data System (ADS)

    McFarlane, Ronald; Hamilton, M. Coreen

    1987-10-01

    A sensor based on fluorescence quenching has been built to detect oxygen activity in gas and water. The sensor consists of a xenon flash bulb as a light source; an excitation wavelength band pass filter; a dichroic beam splitter; collimating and focussing lenses; a plastic clad silica (PCS) rod with the fluorophore immobilized at the tip of it; an emission wavelength band pass filter; a photomultiplier tube (PMT); a monitor PIN photodiode detector; and interface electronics to couple a computer to the rest of the sensor. The device demonstrates a reversible change in fluorescence quenching for changes in oxygen activity. The fluorescence signal seen by the PMT varies over a factor of 3, being highest at 0 oxygen activity and lowest at atmospheric oxygen activity. The device exhibits a 63 % response time of less than 1 second for gases and less than 10 seconds for oxygen dissolved in water. The noise floor of the sensor is approximately 1%. The present embodiment of the device was designed to allow the sensor to operate in the marine environment. The optical components, computer, batteries, and power supply circuitry are mounted on a rack that is enclosed in a pressure housing. The immobilized fluorophore is exposed to sea water. The light travels along the PCS rod, through a pressure seal, to the rest of the system. Present investigations are centered around long term stability of the fluorophore and constituents of the real ocean that will interfere with the quenching mechanism.

  4. The toolbox of fluorescence standards: flexible calibration tools for the standardization of fluorescence-based measurements

    NASA Astrophysics Data System (ADS)

    Resch-Genger, Ute; Hoffmann, K.; Würth, C.; Behnke, T.; Hoffmann, A.; Pfeifer, D.; Engel, A.

    2010-04-01

    To improve the reliability of fluorescence data in the life and material sciences and to enable accreditation of fluorescence techniques, standardization concepts are required that guarantee and improve the comparability of fluorescence measurements. At the core of such concepts are simple and evaluated fluorescence standards for the consideration of instrument-specific spectral and intensity distortions of measured signals and for instrument performance validation (IPV). Similarly in need are fluorescence intensity standards for the quantification from measured intensities and for signal referencing, thereby accounting for excitation light-induced intensity fluctuations. These standards should be preferably certified, especially for use in regulated areas like medical diagnostics. This encouraged us to develop liquid and solid standards for different fluorescence parameters and techniques for use under routine measurement conditions in different formates. Special emphasis was dedicated to the determination and control of the spectral responsivity of detection systems, wavelength accuracy, homogeneity of illumination, and intensity referencing for e.g. spectrofluorometers, fluorescence sensors and confocal laser scanning fluorescence microscopes. Here, we will present design concepts and examples for mono- and multifunctional fluorescence standards that provide traceability to radiometric units and present a first step towards a toolbox of standards.

  5. A capillary-based probe for in situ detection of enhanced fluorescence signals

    NASA Astrophysics Data System (ADS)

    Long, F.; Xiao, R.; Zhu, A. N.; Shi, H. C.; Wang, S. Q.

    2013-07-01

    A simple, compact, and high sensitivity capillary-based probe for the in situ detection of fluorescence signals with high sensitivity is demonstrated. A home-made single-multi-mode fiber coupler that is coaxially aligned with the capillary-based probe provides for the transmission of excitation light and the collection and transmission of fluorescence. We propose a conceptually straightforward theoretical model to optimize the factors affecting the fluorescence-capture capability of the capillary-based probe. The fluorescence signal detected by fiber-optic spectroscopy non-linearly increases with the length of the capillary-based probe. In addition, the thicker the capillary tube wall is, the less the fluorescence signals determined are. The performance of the proposed probe is evaluated experimentally by measuring the fluorescence spectra of Cy5.5 dye and blue-green algae. The experimental results show that the proposed probe provides more than a ten-fold increase in fluorescence signal compared with direct measurements by a flat-tipped multi-mode fiber probe. The advantages of the capillary-based probe, which include its simple and compact structure, excellent light collection efficiency, requirement of small sample volume, and recoverability of samples, allow its wide application to in situ detection in the medical, forensic, biological, geological, and environmental fields with high sensitivity.

  6. Site-specific analysis of protein hydration based on unnatural amino acid fluorescence.

    PubMed

    Amaro, Mariana; Brezovsk, Jan; Kov?ov, Silvia; Skora, Jan; Bedn?, David; N?mec, Vclav; Likov, Veronika; Kurumbang, Nagendra Prasad; Beerens, Koen; Chaloupkov, Radka; Paruch, Kamil; Hof, Martin; Damborsk, Ji?

    2015-04-22

    Hydration of proteins profoundly affects their functions. We describe a simple and general method for site-specific analysis of protein hydration based on the in vivo incorporation of fluorescent unnatural amino acids and their analysis by steady-state fluorescence spectroscopy. Using this method, we investigate the hydration of functionally important regions of dehalogenases. The experimental results are compared to findings from molecular dynamics simulations. PMID:25815779

  7. Fluorescent Protein Based FRET Pairs with Improved Dynamic Range for Fluorescence Lifetime Measurements

    PubMed Central

    George Abraham, Bobin; Sarkisyan, Karen S.; Mishin, Alexander S.; Santala, Ville; Tkachenko, Nikolai V.; Karp, Matti

    2015-01-01

    Fluorescence Resonance Energy Transfer (FRET) using fluorescent protein variants is widely used to study biochemical processes in living cells. FRET detection by fluorescence lifetime measurements is the most direct and robust method to measure FRET. The traditional cyan-yellow fluorescent protein based FRET pairs are getting replaced by green-red fluorescent protein variants. The green-red pair enables excitation at a longer wavelength which reduces cellular autofluorescence and phototoxicity while monitoring FRET. Despite the advances in FRET based sensors, the low FRET efficiency and dynamic range still complicates their use in cell biology and high throughput screening. In this paper, we utilized the higher lifetime of NowGFP and screened red fluorescent protein variants to develop FRET pairs with high dynamic range and FRET efficiency. The FRET variations were analyzed by proteolytic activity and detected by steady-state and time-resolved measurements. Based on the results, NowGFP-tdTomato and NowGFP-mRuby2 have shown high potentials as FRET pairs with large fluorescence lifetime dynamic range. The in vitro measurements revealed that the NowGFP-tdTomato has the highest Frster radius for any fluorescent protein based FRET pairs yet used in biological studies. The developed FRET pairs will be useful for designing FRET based sensors and studies employing Fluorescence Lifetime Imaging Microscopy (FLIM). PMID:26237400

  8. Simple non-fluorescent polarity labeling of microtubules for molecular motor assays.

    PubMed

    Soppina, Virupakshi; Rai, Arpan; Mallik, Roop

    2009-06-01

    Transport of intracellular organelles along the microtubule cytoskeleton occurs in a bidirectional manner due to opposing activity of microtubule-associated motor proteins of the kinesin and dynein families. Regulation of this opposing activity and the resultant motion is believed to generate a polarized distribution of many organelles within the cell. The bidirectional motion can be reconstituted on in vitro assembled microtubules using organelles extracted from cells. This provides an opportunity to understand the regulation of intracellular transport through quantitative analysis of the motion of organelles in a controlled environment. Such analysis requires the use of polarity-labeled microtubules to resolve the plus and minus components of bidirectional motion. However, existing methods of in vitro microtubule polarity labeling are unsuitable for high-resolution recording of motion. Here we present a simple and reliable method that uses avidin-coated magnetic beads to prepare microtubules labeled at the minus end. The microtubule polarity can be identified without any need for fluorescence excitation. We demonstrate video-rate high-resolution imaging of single cellular organelles moving along plus and minus directions on labeled microtubules. Quantitative analysis of this motion indicates that these organelles are likely to be driven by multiple dynein motors in vivo. PMID:19594454

  9. Simple Method for Fluorescence DNA In Situ Hybridization to Squashed Chromosomes

    PubMed Central

    Larracuente, Amanda M.; Ferree, Patrick M.

    2015-01-01

    DNA in situ hybridization (DNA ISH) is a commonly used method for mapping sequences to specific chromosome regions. This approach is particularly effective at mapping highly repetitive sequences to heterochromatic regions, where computational approaches face prohibitive challenges. Here we describe a streamlined protocol for DNA ISH that circumvents formamide washes that are standard steps in other DNA ISH protocols. Our protocol is optimized for hybridization with short single strand DNA probes that carry fluorescent dyes, which effectively mark repetitive DNA sequences within heterochromatic chromosomal regions across a number of different insect tissue types. However, applications may be extended to use with larger probes and visualization of single copy (non-repetitive) DNA sequences. We demonstrate this method by mapping several different repetitive sequences to squashed chromosomes from Drosophila melanogaster neural cells and Nasonia vitripennis spermatocytes. We show hybridization patterns for both small, commercially synthesized probes and for a larger probe for comparison. This procedure uses simple laboratory supplies and reagents, and is ideal for investigators who have little experience with performing DNA ISH. PMID:25591075

  10. Simple and sensitive determination of five quinolones in food by liquid chromatography with fluorescence detection.

    PubMed

    Ramos, Macarena; Aranda, Angela; Garcia, Elena; Reuvers, Thea; Hooghuis, Henny

    2003-06-15

    A simple and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the determination of five different quinolones: enrofloxacin, ciprofloxacin, sarafloxacin, oxolinic acid and flumequine in pork and salmon muscle. The method includes one extraction and clean-up step for the five quinolones together which are detected in two separated HPLC runs by means of their fluorescence. The proposed analytical method involves homogenizing of the tissue sample with 0.05 M phosphate buffer, pH 7.4 and clean-up by Discovery DS-18 cartridges. For chromatographic separation a Symmetry C(18) column is used in two different runs: (1) ciprofloxacin, enrofloxacin and sarafloxacin with acetonitrile-0.02 M phosphate buffer pH 3.0 (18:82) as mobile phase and the detector at excitation wavelength: 280 nm and emission wavelength 450 nm; and (2) oxolinic acid and flumequine with acetonitrile-0.02 M phosphate buffer pH 3.0 (34:66) as mobile phase and excitation wavelength: 312 nm and emission wavelength: 366 nm. Detection limit was as low as 5 ng g(-1), except for sarafloxacin which had a limit of 10 ng g(-1). Standard curves using blank muscle tissues spiked at different levels showed a good linear correlation coefficient, r(2) higher than 0.999 for all quinolones. PMID:12742128

  11. A highly selective quinoline-based fluorescent sensor for Zn(II)

    NASA Astrophysics Data System (ADS)

    Kim, Hyun; Kang, Juhye; Kim, Kyung Beom; Song, Eun Joo; Kim, Cheal

    2014-01-01

    A quinoline-based simple receptor (bis(2-quinolinylmethyl)benzylamine = 1) as a Zn2+ selective fluorescent chemosensor showed a large fluorescent enhancement with a blue shift in the presence of Zn2+ which is attributed to a chelation enhanced fluorescence (CHEF) effect with inhibition of a photoinduced electron transfer (PET) process of 1. In particular, this receptor could clearly distinguish Zn2+ from Cd2+. The binding mode of 1 and Zn2+ was found to be a 1:1 and confirmed by Job plot, 1H NMR titration and ESI-mass spectrometry analysis.

  12. A highly selective quinoline-based fluorescent sensor for Zn(II).

    PubMed

    Kim, Hyun; Kang, Juhye; Kim, Kyung Beom; Song, Eun Joo; Kim, Cheal

    2014-01-24

    A quinoline-based simple receptor (bis(2-quinolinylmethyl)benzylamine = 1) as a Zn(2+) selective fluorescent chemosensor showed a large fluorescent enhancement with a blue shift in the presence of Zn(2+) which is attributed to a chelation enhanced fluorescence (CHEF) effect with inhibition of a photoinduced electron transfer (PET) process of 1. In particular, this receptor could clearly distinguish Zn(2+) from Cd(2+). The binding mode of 1 and Zn(2+) was found to be a 1:1 and confirmed by Job plot, (1)H NMR titration and ESI-mass spectrometry analysis. PMID:24161851

  13. Simple process of hybrid white quantum dot/organic light-emitting diodes by using quantum dot plate and fluorescence

    NASA Astrophysics Data System (ADS)

    Lee, Ho Won; Lee, Ki-Heon; Lee, Jae Woo; Kim, Jong-Hoon; Yang, Heesun; Kim, Young Kwan

    2015-02-01

    In this work, the simple process of hybrid quantum dot (QD)/organic light-emitting diode (OLED) was proposed to apply a white illumination light by using QD plate and organic fluorescence. Conventional blue fluorescent OLEDs were firstly fabricated and then QD plates of various concentrations, which can be controlled of UV-vis absorption and photoluminescence spectrum, were attached under glass substrate of completed blue devices. The suggested process indicates that we could fabricate the white device through very simple process without any deposition of orange or red organic emitters. Therefore, this work would be demonstrated that the potential simple process for white applications can be applied and also can be extended to additional research on light applications.

  14. Sensitive turn-on fluorescent detection of melamine based on fluorescence resonance energy transfer.

    PubMed

    Guo, Liangqia; Zhong, Jianhai; Wu, Jinmei; Fu, FengFu; Chen, Guonan; Chen, Yongxuan; Zheng, Xiaoyan; Lin, Song

    2011-04-21

    We here report a novel fluorescent method for the detection of melamine based on the high fluorescence quenching ability of gold nanoparticles. The fluorescence was significantly quenched via fluorescence resonance energy transfer when fluorescein molecules were attached to the surface of gold nanoparticles by electrostatic interaction. Upon addition of melamine, the fluorescence was enhanced due to the competitive adsorption of gold nanoparticles between melamine and fluorescein. Under the optimum conditions, the fluorescence enhancement efficiency [(I-I(0))/I(0)] showed a linear relationship with the concentration of melamine in the range of 1.0 × 10(-7) mol L(-1)~4.0 × 10(-6) mol L(-1), and the detection limit was calculated to be 1.0 × 10(-9) mol L(-1). The proposed method showed several advantages such as high sensitivity, short analysis time, low cost and ease of operation. PMID:21359305

  15. Quantum dots-based label-free fluorescence sensor for sensitive and non-enzymatic detection of caffeic acid.

    PubMed

    Xiang, Xia; Shi, Jianbin; Huang, Fenghong; Zheng, Mingming; Deng, Qianchun

    2015-08-15

    We have developed a label-free fluorescence sensor for caffeic acid (CA) by the use of CdTe:Zn(2+) quantum dots (CdTe:Zn(2+) QDs) as an output signal. The principle of sensor is based on the fluorescence quenching and binding properties of Fe(2+) toward QDs and CA, respectively. To provide a fluorescence turn-on mode for CA detection, Fe(2+) is first mixed with QDs solution, leading to a low fluorescence emission. With the addition of CA, the fluorescence of QDs is recovered due to the strong binding interaction between CA and Fe(2+). Thus, a QDs-based label-free fluorescence sensor, designed in a simple mix-and-detect format, is established for CA detection. This study demonstrated here not only offers simple, sensitive and non-enzymatic detection method for CA, but also brings to light a new application of QDs in the food analysis. PMID:25966400

  16. A simple and sensitive UFLC-fluorescence method for endocrine disrupters determination in marine waters.

    PubMed

    Lisboa, Normando S; Fahning, Cristiane S; Cotrim, Gabriel; dos Anjos, Jeancarlo P; de Andrade, Jailson B; Hatje, Vanessa; da Rocha, Gisele O

    2013-12-15

    The present study proposes a fast and simple analytical methodology employing C18 SPE cartridges (for preconcentration and clean-up), and a ultra-fast liquid chromatography coupled to fluorescence detector (UFLC-FLD) for determination of the following endocrine disrupters (ED): bisphenol A (BPA), 4-n-nonylphenol (4NNP), 4-n-octylphenol (4NOP), 4-t-octylphenol (4TOP), estriol (E3), estrone (E1), 17?-estradiol (E2) and 17?-ethynylestradiol (EE2) in seawater. The proposed method was developed, optimized and validated. Separation was done by a total running time of 10 min in a Shim-pack XR-ODS C-18 (2.0 mm ID 50 mm) chromatographic column, mobile phases were acetonitrile/ultra-pure water under gradient programming; eluent flow rate at 0.120 mL min(-1); column temperature set at 60 C; emission wavelength of 306 nm and excitation wavelength of 280 nm. The method was validated through assessment of the following parameters: linear range, linearity, selectiveness, precision, recovery test, limit of detection (LOD), and limit of quantification (LOQ). Recoveries ranged from 91% (for EE2) to 104% (for 4NNP) and also was found a suitable repeatability (RSD <4.5%) for all considered compounds. LOD and LOQ ranged from 2.0 ng L(-1) (EE2) to 23 ng L(-1) (E1) and 9.3 ng L(-1) (EE2) to 96 ng L(-1) (E1), respectively. The analytical method using SPE UFLC-FLD was applied to seawater samples collected from Todos os Santos Bay (BTS), Brazil to determine the concentration of eight ED. PMID:24209326

  17. Fluorescent nanoparticles based on AIE fluorogens for bioimaging

    NASA Astrophysics Data System (ADS)

    Yan, Lulin; Zhang, Yan; Xu, Bin; Tian, Wenjing

    2016-01-01

    Fluorescent nanoparticles (FNPs) have recently attracted increasing attention in the biomedical field because of their unique optical properties, easy fabrication and outstanding performance in imaging. Compared with conventional molecular probes including small organic dyes and fluorescent proteins, FNPs based on aggregation-induced emission (AIE) fluorogens have shown significant advantages in tunable emission and brightness, good biocompatibility, superb photo- and physical stability, potential biodegradability and facile surface functionalization. In this review, we summarize the latest advances in the development of fluorescent nanoparticles based on AIE fluorogens including polymer nanoparticles and silica nanoparticles over the past few years, and the various biomedical applications based on these fluorescent nanoparticles are also elaborated.

  18. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE: INDUCED BY RADIATION, CHEMICALS AND ENZYMES

    EPA Science Inventory

    A simple and rapid assay to detect DNA damage is reported. This assay is based on the ability of certain dyes to fluoresce upon intercalation with dsDNA. Damage caused by ultraviolet (UV) radiation, chemicals or restriction enzymes is detected using this assay. UV radiation at...

  19. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE INDUCED BY RADIATION, CHEMICAL MUTAGENS AND ENZYMES

    EPA Science Inventory

    A simple and rapid assay to detect DNA damage is reported. This novel assay is based on changes in melting/annealing behavior and facilitated using certain dyes that increase their fluorescence upon association with double stranded (ds)DNA. Damage caused by ultraviolet (UV) ra...

  20. A FLUORESCENCE BASED ASSAY FOR DNA DAMAGE INDUCED BY STYRENE OXIDE

    EPA Science Inventory

    A rapid and simple assay to detect DNA damage to calf thymus DNA caused by styrene oxide (SO) is reported. This assay is based on changes observed in the melting and annealing behavior of the damaged DNA. The melting annealing process was monitored using a fluorescence indicat...

  1. Development of Highly Fluorescent Materials Based on Thiophenylimidazole Dyes

    NASA Technical Reports Server (NTRS)

    Santos, Javier; Bu, Xiu R.; Mintz, Eric A.; Meador, Michael A. (Technical Monitor)

    2000-01-01

    Organic fluorescent materials are expected to find many potential applications in optical devices and photo-functionalized materials. Although many investigations have been focused on heterocyclic compounds such as coumarins, bipyridines, rhodamines, and pyrrole derivatives, little is known for fluorescent imidazole materials. We discovered that one particular class of imidazole derivatives is highly fluorescent. A series of monomeric and polymeric based fluorescent dyes were prepared containing a thiophene unit at the second position of the imidazole ring. Dependence of fluorescence efficiency on parameters such as solvent polarity and substituent groups has been investigated. It was found that a formyl group at the 2-position of the thiophene ring dramatically enhance fluorescence properties. Ion recognition probes indicated their potential as sensor materials. These fluorophores have flexibility for introduction of versatile substituent groups that could improve the fluorescence efficiency and sensor properties.

  2. Fluorescent-protein-based probes: general principles and practices.

    PubMed

    Ai, Hui-Wang

    2015-01-01

    An important application of fluorescent proteins is to derive genetically encoded fluorescent probes that can actively respond to cellular dynamics such as pH change, redox signaling, calcium oscillation, enzyme activities, and membrane potential. Despite the large diverse group of fluorescent-protein-based probes, a few basic principles have been established and are shared by most of these probes. In this article, the focus is on these general principles and strategies that guide the development of fluorescent-protein-based probes. A few examples are provided in each category to illustrate the corresponding principles. Since these principles are quite straightforward, others may adapt them to create fluorescent probes for their own interest. Hopefully, the development of the ever-growing family of fluorescent-protein-based probes will no longer be limited to a small number of laboratories specialized in senor development, leading to the situation that biological studies will be bettered assisted by genetically encoded sensors. PMID:25326886

  3. A planarized 9-phenylanthracene: a simple electron-donating building block for fluorescent materials.

    PubMed

    Iwahara, Hideaki; Kushida, Tomokatsu; Yamaguchi, Shigehiro

    2016-01-01

    A 9-phenylanthracene, constrained in a coplanar fashion by two methylene tethers, was synthesized and its optoelectronic properties were investigated. The rigid planarization increased its electron-donating character and induced a bathochromic shift of its absorption, as well as an increased molar absorption coefficient and intense fluorescence. These properties render this compound a promising building block for fluorescent materials. PMID:26515716

  4. Simple and rapid determination of homozygous transgenic mice via in vivo fluorescence imaging

    PubMed Central

    Li, Wei; Xiao, Gaofang; Li, Yanqing; Xie, Raoying; Huang, Hailu; Zhong, Lin; Wu, Qinghong; Wang, Wanshan; Huang, Wenhua; Yao, Kaitai; Xiao, Dong; Sun, Yan

    2015-01-01

    Setting up breeding programs for transgenic mouse strains require to distinguish homozygous from the heterozygous transgenic animals. The combinational use of the fluorescence reporter transgene and small animal in-vivo imaging system might allow us to rapidly and visually determine the transgenic mice homozygous for transgene(s) by the in vivo fluorescence imaging. RLG, RCLG or Rm17LG transgenic mice ubiquitously express red fluorescent protein (RFP). To identify homozygous RLG transgenic mice, whole-body fluorescence imaging for all of newborn F2-generation littermates produced by mating of RFP-positive heterozygous transgenic mice (F1-generation) derived from the same transgenic founder was performed. Subsequently, the immediate data analysis of the in vivo fluorescence imaging was carried out, which greatly facilitated us to rapidly and readily distinguish RLG transgenic individual(s) with strong fluorescence from the rest of F2-generation littermates, followed by further determining this/these RLG individual(s) showing strong fluorescence to be homozygous, as strongly confirmed by mouse mating. Additionally, homozygous RCLG or Rm17LG transgenic mice were also rapidly and precisely distinguished by the above-mentioned optical approach. This approach allowed us within the shortest time period to obtain 10, 8 and 2 transgenic mice homozygous for RLG, RCLG and Rm17LG transgene, respectively, as verified by mouse mating, indicating the practicality and reliability of this optical method. Taken together, our findings fully demonstrate that the in vivo fluorescence imaging offers a visual, rapid and reliable alternative method to the traditional approaches (i.e., mouse mating and real-time quantitative PCR) in identifying homozygous transgenic mice harboring fluorescence reporter transgene under the control of a ubiquitous promoter in the situation mentioned in this study. PMID:26472024

  5. Simple and rapid determination of homozygous transgenic mice via in vivo fluorescence imaging.

    PubMed

    Lin, Xiaolin; Jia, Junshuang; Qin, Yujuan; Lin, Xia; Li, Wei; Xiao, Gaofang; Li, Yanqing; Xie, Raoying; Huang, Hailu; Zhong, Lin; Wu, Qinghong; Wang, Wanshan; Huang, Wenhua; Yao, Kaitai; Xiao, Dong; Sun, Yan

    2015-11-17

    Setting up breeding programs for transgenic mouse strains require to distinguish homozygous from the heterozygous transgenic animals. The combinational use of the fluorescence reporter transgene and small animal in-vivo imaging system might allow us to rapidly and visually determine the transgenic mice homozygous for transgene(s) by the in vivo fluorescence imaging. RLG, RCLG or Rm17LG transgenic mice ubiquitously express red fluorescent protein (RFP). To identify homozygous RLG transgenic mice, whole-body fluorescence imaging for all of newborn F2-generation littermates produced by mating of RFP-positive heterozygous transgenic mice (F1-generation) derived from the same transgenic founder was performed. Subsequently, the immediate data analysis of the in vivo fluorescence imaging was carried out, which greatly facilitated us to rapidly and readily distinguish RLG transgenic individual(s) with strong fluorescence from the rest of F2-generation littermates, followed by further determining this/these RLG individual(s) showing strong fluorescence to be homozygous, as strongly confirmed by mouse mating. Additionally, homozygous RCLG or Rm17LG transgenic mice were also rapidly and precisely distinguished by the above-mentioned optical approach. This approach allowed us within the shortest time period to obtain 10, 8 and 2 transgenic mice homozygous for RLG, RCLG and Rm17LG transgene, respectively, as verified by mouse mating, indicating the practicality and reliability of this optical method. Taken together, our findings fully demonstrate that the in vivo fluorescence imaging offers a visual, rapid and reliable alternative method to the traditional approaches (i.e., mouse mating and real-time quantitative PCR) in identifying homozygous transgenic mice harboring fluorescence reporter transgene under the control of a ubiquitous promoter in the situation mentioned in this study. PMID:26472024

  6. Phytoplankton photocompensation from space-based fluorescence measurements

    NASA Astrophysics Data System (ADS)

    Morrison, J. Ruairidh; Goodwin, Deborah S.

    2010-03-01

    Recent satellite-derived observations linked global scale phytoplankton fluorescence variability with iron stress and hinted at photophysiological responses associated with changing light levels. These photocompensation reactions, the sum of photoacclimation and photoadaptation, were examined with climatological data for the Gulf of Maine. Significant seasonal variability was observed in the fluorescence quantum yield that was unrelated to patterns of biomass. Up to 89% of the variability in the fluorescence quantum yield was explained by a physiology-based photocompensation model. Spatial variability in seasonal patterns was associated with differing hydrodynamic regimes. This variability in the quantum yield demonstrates that satellite-based fluorescence is inappropriate for phytoplankton biomass determinations. More importantly, the work presented here provides the modeling foundation for fluorescence-based investigations of temporal and spatial variability in phytoplankton physiology associated with growth irradiance. These space-based physiological observations have the potential to decrease uncertainties in future ocean color derived primary productivity estimates.

  7. Disposable nitrate-selective optical sensor based on fluorescent dye

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A simple, disposable thin-film optical nitrate sensor was developed. The sensor was fabricated by applying a nitrate-selective polymer membrane on the surface of a thin polyester film. The membrane was composed of polyvinylchloride (PVC), plasticizer, fluorescent dye, and nitrate-selective ionophore...

  8. Characterization of Flavin-Based Fluorescent Proteins: An Emerging Class of Fluorescent Reporters

    PubMed Central

    Mukherjee, Arnab; Schroeder, Charles M.

    2013-01-01

    Fluorescent reporter proteins based on flavin-binding photosensors were recently developed as a new class of genetically encoded probes characterized by small size and oxygen-independent maturation of fluorescence. Flavin-based fluorescent proteins (FbFPs) address two major limitations associated with existing fluorescent reporters derived from the green fluorescent protein (GFP)namely, the overall large size and oxygen-dependent maturation of fluorescence of GFP. However, FbFPs are at a nascent stage of development and have been utilized in only a handful of biological studies. Importantly, a full understanding of the performance and properties of FbFPs as a practical set of biological probes is lacking. In this work, we extensively characterize three FbFPs isolated from Pseudomonas putida, Bacillus subtilis, and Arabidopsis thaliana, using in vitro studies to assess probe brightness, oligomeric state, maturation time, fraction of fluorescent holoprotein, pH tolerance, redox sensitivity, and thermal stability. Furthermore, we validate FbFPs as stable molecular tags using in vivo studies by constructing a series of FbFP-based transcriptional constructs to probe promoter activity in Escherichia coli. Overall, FbFPs show key advantages as broad-spectrum biological reporters including robust pH tolerance (411), thermal stability (up to 60C), and rapid maturation of fluorescence (<3 min.). In addition, the FbFP derived from Arabidopsis thaliana (iLOV) emerged as a stable and nonperturbative reporter of promoter activity in Escherichia coli. Our results demonstrate that FbFP-based reporters have the potential to address key limitations associated with the use of GFP, such as pH-sensitive fluorescence and slow kinetics of fluorescence maturation (1040 minutes for half maximal fluorescence recovery). From this view, FbFPs represent a useful new addition to the fluorescent reporter protein palette, and our results constitute an important framework to enable researchers to implement and further engineer improved FbFP-based reporters with enhanced brightness and tighter flavin binding, which will maximize their potential benefits. PMID:23741385

  9. Mosaic-Detector-Based Fluorescence Spectral Imager

    NASA Technical Reports Server (NTRS)

    Son, Kyung-Ah; Moon, Jeong

    2007-01-01

    A battery-powered, pen-sized, portable instrument for measuring molecular fluorescence spectra of chemical and biological samples in the field has been proposed. Molecular fluorescence spectroscopy is among the techniques used most frequently in laboratories to analyze compositions of chemical and biological samples. Heretofore, it has been possible to measure fluorescence spectra of molecular species at relative concentrations as low as parts per billion (ppb), with a few nm spectral resolution. The proposed instrument would include a planar array (mosaic) of detectors, onto which a fluorescence spectrum would be spatially mapped. Unlike in the larger laboratory-type molecular fluorescence spectrometers, mapping of wavelengths to spatial positions would be accomplished without use of relatively bulky optical parts. The proposed instrument is expected to be sensitive enough to enable measurement of spectra of chemical species at relative concentrations <1 ppb, with spectral resolution that could be tailored by design to be comparable to a laboratory molecular fluorescence spectrometer. The proposed instrument (see figure) would include a button-cell battery and a laser diode, which would generate the monochromatic ultraviolet light needed to excite fluorescence in a sample. The sample would be held in a cell bounded by far-ultraviolet-transparent quartz or optical glass. The detector array would be, more specifically, a complementary metal oxide/ semiconductor or charge-coupled- device imaging photodetector array, the photodetectors of which would be tailored to respond to light in the wavelength range of the fluorescence spectrum to be measured. The light-input face of the photodetector array would be covered with a matching checkerboard array of multilayer thin film interference filters, such that each pixel in the array would be sensitive only to light in a spectral band narrow enough so as not to overlap significantly with the band of an adjacent pixel. The wavelength interval between adjacent pixels (and, thus, the spectral resolution) would typically be chosen by design to be approximately equal to the width of the total fluorescence wavelength range of interest divided by the number of pixels. The unitary structure comprising the photodetector array overlaid with the matching filter array would be denoted a hyperspectral mosaic detector (HMD) array.

  10. A fluorescence-based assay for monitoring helicase activity.

    PubMed

    Raney, K D; Sowers, L C; Millar, D P; Benkovic, S J

    1994-07-01

    A continuous fluorescence-based assay is described for measuring helicase-mediated unwinding of duplex DNA. The assay utilizes an oligonucleotide substrate containing the fluorescent adenine analog, 2-aminopurine, at regular intervals. 2-Aminopurine forms a Watson-Crick-type base pair with thymine and does not distort normal B-form DNA. Fluorescence of the 2-aminopurines within this oligonucleotide is quenched 2-fold upon its hybridization to a complementary strand. Unwinding of this substrate by the T4 dda helicase restores the fluorescence of the 2-aminopurines and is easily followed using stopped-flow or steady-state fluorescence spectroscopy. The flourescence-based assay provides rate data comparable to that obtained from conventional discontinuous assays using labeled substrates and additionally furnishes a means for following a single turnover. This assay should prove useful for defining the mechanism by which helicases unwind duplex DNA. PMID:8022830

  11. Radiative transport-based frequency-domain fluorescence tomography

    NASA Astrophysics Data System (ADS)

    Joshi, Amit; Rasmussen, John C.; Sevick-Muraca, Eva M.; Wareing, Todd A.; McGhee, John

    2008-04-01

    We report the development of radiative transport model-based fluorescence optical tomography from frequency-domain boundary measurements. The coupled radiative transport model for describing NIR fluorescence propagation in tissue is solved by a novel software based on the established Attila particle transport simulation platform. The proposed scheme enables the prediction of fluorescence measurements with non-contact sources and detectors at a minimal computational cost. An adjoint transport solution-based fluorescence tomography algorithm is implemented on dual grids to efficiently assemble the measurement sensitivity Jacobian matrix. Finally, we demonstrate fluorescence tomography on a realistic computational mouse model to locate nM to M fluorophore concentration distributions in simulated mouse organs.

  12. Eco-friendly carbon-nanodot-based fluorescent paints for advanced photocatalytic systems

    NASA Astrophysics Data System (ADS)

    Young Park, So; Uk Lee, Hyun; Lee, Young-Chul; Choi, Saehae; Hyun Cho, Dae; Sik Kim, Hee; Bang, Sunghee; Seo, Soonjoo; Chang Lee, Soon; Won, Jonghan; Son, Byung-Chul; Yang, Mino; Lee, Jouhahn

    2015-07-01

    Fluorescent carbon nanomaterials, especially zero-dimensional (0D) carbon nanodots (CDs), are widely used in broad biological and optoelectronic applications. CDs have unique characteristics such as strong fluorescence, biocompatibility, sun-light response, and capability of mass-production. Beyond the previous green CD obtained from harmful natural substances, we report a new type of fluid-based fluorescent CD paints (C-paints) derived from polyethylene glycol (PEG; via simple ultrasound irradiation at room temperatures) and produced in quantum yields of up to ~14%. Additionally, C-paints possess a strong, UV- and visible-light-responsive photoluminescent (PL) property. Most especially, C-paints, by incorporation into a photocatalytic system, show additional roles in the emission of fluorescent light for activation of TiO2 nanoparticles (NPs) and the resultant detoxification of most organic dyes, thus further enabling embarkation in advanced water purification.

  13. Fluorescence-based resource for semi-automated genomic analyses

    SciTech Connect

    Kiser, M.B.; Dragwa, C.; Jedlicka, A.E.

    1994-09-01

    To facilitate the practical application of highly efficient semi-automated methods for general application in genomic analyses, we have developed a fluorescence-based marker resource. Ninety highly polymorphic simple tandem repeat markers were combined to provide a rapid, accurate, and highly efficient initial genome-wide screening system. These markers are spaced on average every 33 recombination units, with a mean heterozygosity of 81% (range 65-94%), covering 22 autosomes and the X and Y chromosomes. Less than 3% of the genome lies beyond 30 cM of the nearest marker. Markers were placed in a vertical ladder that we have termed a SET according to the size of the PCR fragments they produce during electrophoresis. Each SET was designed to avoid overlap between loci during gel separations to assure accuracy when scoring genotypes. We have constructed 15 SETS of markers. Three SETS, each labelled with one of three fluors, were combined into what we have termed a GROUP, which is co-electrophoresed with internal size standards that are labelled with a fourth flour. Five GROUPS of markers were assembled that contain a total of 15 SETS of markers. Each GROUP cover 18 regions of the genome that can be detected simultaneously, since this genomic analysis system is fully compatible with automated fragment analyzers using simultaneous four-color fluorescence-based detection systems. This allows for multiplex detection and a throughput of 1,944 genotypes daily per instrument. This system will be highly beneficial in a number of clinical and research applications including: linkage, cancer genetics, forensics, and cytogenetics.

  14. A sensitive fluorescent sensor for selective determination of dichlorvos based on the recovered fluorescence of carbon dots-Cu(II) system.

    PubMed

    Hou, Juying; Dong, Guangjuan; Tian, Zhengbin; Lu, Jiutian; Wang, Qianqian; Ai, Shiyun; Wang, Minglin

    2016-07-01

    In this paper, a simple and sensitive fluorescent sensor for dichlorvos was first constructed based on carbon dots-Cu(II) system. These carbon dots were obtained by simple hydrothermal reaction of feather. The fluorescence of these carbon dots can be selectively quenched by Cu(2+) ion. When acetylcholinesterase and acetylthiocholine were introduced into the system, thiocholine came into being, which can react with Cu(2+) ion and restore the fluorescence of the system. The reaction mechanism between Cu(2+) ion and thiocholine was confirmed by X-ray photoelectron spectroscopy. As one kind of acetylcholinesterase inhibitor, organophosphorus pesticides can be detected based on this sensing system. As an example of organophosphorus pesticides, dichlorvos was detected with a linear range of 6.0×10(-9)-6.0×10(-8)M. This sensing system has been successfully used for the analysis of cabbage and fruit juice samples. PMID:26920268

  15. A simple integrated microfluidic device for the multiplexed fluorescence-free detection of Salmonella enterica.

    PubMed

    Strachan, Briony C; Sloane, Hillary S; Houpt, Eric; Lee, Jacob C; Miranian, Daniel C; Li, Jingyi; Nelson, Daniel A; Landers, James P

    2016-02-01

    Rapid, inexpensive and simplistic nucleic acid testing (NAT) is pivotal in delivering biotechnology solutions at the point-of-care (POC). We present a poly(methylmethacrylate) (PMMA) microdevice where on-board infrared-mediated PCR amplification is seamlessly integrated with a particle-based, visual DNA detection for specific detection of bacterial targets in less than 35 minutes. Fluidic control is achieved using a capillary burst valve laser-ablated in a novel manner to confine the PCR reagents to a chamber during thermal cycling, and a manual torque-actuated pressure system to mobilize the fluid from the PCR chamber to the detection reservoir containing oligonucleotide-adducted magnetic particles. Interaction of amplified products specific to the target organism with the beads in a rotating magnetic field allows for near instantaneous (<30 s) detection based on hybridization-induced aggregation (HIA) of the particles and simple optical analysis. The integration of PCR with this rapid, sequence-specific DNA detection method on a single microdevice presents the possibility of creating POC NAT systems that are low cost, easy-to-use, and involve minimal external hardware. PMID:26658961

  16. Molecular spies for bioimaging--fluorescent protein-based probes.

    PubMed

    Miyawaki, Atsushi; Niino, Yusuke

    2015-05-21

    Convergent advances in optical imaging and genetic engineering have fueled the development of new technologies for biological visualization. Those technologies include genetically encoded indicators based on fluorescent proteins (FPs) for imaging ions, molecules, and enzymatic activities "to spy on cells," as phrased by Roger Tsien, by sneaking into specific tissues, cell types, or subcellular compartments, and reporting on specific intracellular activities. Here we review the current range of unimolecular indicators whose working principle is the conversion of a protein conformational change into a fluorescence signal. Many of the indicators have been developed from fluorescence resonance energy transfer- and single-FP-based approaches. PMID:26000848

  17. A simple-structured acridine derivative as a fluorescent enhancement chemosensor for the detection of Pd2+ in aqueous media.

    PubMed

    Zhou, Yanmei; Huang, Qi; Zhang, Qingyou; Min, Yinghao; Wang, Enze

    2015-02-25

    4,5-Bis(hydroxymethyl) acridine (sensor 1) has been discovered and synthesized as a simple-structured Pd(2+) fluorescent probe. Sensor 1 showed highly selective recognition toward Pd(2+) over other examined metal ions in aqueous solution. Under the optimized condition, fluorescence intensity was linearly proportional to the concentration of Pd(2+) in the 0-1 ?M concentration range with detection limits of 0.021 ?M. The EDTA-adding and stoichiometry experiments indicated that sensor 1 was a reversible chemosensor for Pd(2+) with a 2:1 ligand/metal complex at neutral pH. Moreover, the sensor 1 was also successfully applied to determination of Pd(2+) in water samples and palladium-containing catalyst, which made it attractive for sensing applications. PMID:25194318

  18. A simple-structured acridine derivative as a fluorescent enhancement chemosensor for the detection of Pd2+ in aqueous media

    NASA Astrophysics Data System (ADS)

    Zhou, Yanmei; Huang, Qi; Zhang, Qingyou; Min, Yinghao; Wang, Enze

    2015-02-01

    4,5-Bis(hydroxymethyl) acridine (sensor 1) has been discovered and synthesized as a simple-structured Pd2+ fluorescent probe. Sensor 1 showed highly selective recognition toward Pd2+ over other examined metal ions in aqueous solution. Under the optimized condition, fluorescence intensity was linearly proportional to the concentration of Pd2+ in the 0-1 ?M concentration range with detection limits of 0.021 ?M. The EDTA-adding and stoichiometry experiments indicated that sensor 1 was a reversible chemosensor for Pd2+ with a 2:1 ligand/metal complex at neutral pH. Moreover, the sensor 1 was also successfully applied to determination of Pd2+ in water samples and palladium-containing catalyst, which made it attractive for sensing applications.

  19. Novel pyrazoline-based selective fluorescent probe for the detection of hydrazine

    NASA Astrophysics Data System (ADS)

    Zheng, Xiao-Xin; Wang, Sheng-Qing; Wang, Hao-Yan; Zhang, Rong-Rong; Liu, Jin-Ting; Zhao, Bao-Xiang

    2015-03-01

    A novel pyrazoline-based fluorescent probe, 2-[4-(3,5-diphenyl-4,5-dihydro-pyrazol-1-yl)-benzylidene]-malononitrile, with a simple structure and low detection limit (6.16 10-6 M) for the detection of hydrazine is designed and synthesized. The probe responds selectively to hydrazine over other molecules with marked fluorescence enhancement. The probe can detect hydrazine effectively at pH 5.0-9.0 with a special emission wavelength at 520 nm. Moreover, the probe can be used to detect hydrazine from variety of natural source water.

  20. Compound parabolic concentrator optical fiber tip for FRET-based fluorescent sensors

    NASA Astrophysics Data System (ADS)

    Ul Hassan, Hafeez; Nielsen, Kristian; Aasmul, Soren; Bang, Ole

    2015-09-01

    The Compound Parabolic Concentrator (CPC) optical fiber tip shape has been proposed for intensity based fluorescent sensors working on the principle of FRET (Frster Resonance Energy Transfer). A simple numerical Zemax model has been used to optimize the CPC tip geometry for a step-index multimode polymer optical fiber for an excitation and emission wavelength of 550 nm and 650nm, respectively. The model suggests an increase of a factor of 1.6 to 4 in the collected fluorescent power for an ideal CPC tip, as compared to the plane-cut fiber tip for fiber lengths between 5 and 45mm.

  1. A ratiometric fluorescent quantum dots based biosensor for organophosphorus pesticides detection by inner-filter effect.

    PubMed

    Yan, Xu; Li, Hongxia; Han, Xiaosong; Su, Xingguang

    2015-12-15

    In this work, we develop a novel and sensitive sensor for the detection of organophosphorus pesticides based on the inner-filter effect (IFE) between gold nanoparticles (AuNPs) and ratiometric fluorescent quantum dots (RF-QDs). The RF-QDs has been designed by hybridizing two differently colored CdTe QDs, in which the red emissive QDs entrapped in the silica sphere acting as the reference signal, and the green emissive QDs covalently attached on the silica surface serving as the response signal.The fluorescence of RF-QDs could be quenched by AuNPs based on IFE. Protamine could effectively turn on the fluorescence due to the electrostatic attraction between protamine and AuNPs. Trypsin can easily hydrolyze protamine, leading to the quench of the fluorescence. Then, the fluorescence could be recovered again by the addition of parathion-methyl (PM) which could inhibit the activity of trypsin. By measuring the fluorescence of RF-QDs, the inhibition efficiency of PM to trypsin activity was evaluated. Under the optimized conditions, the inhibition efficiency was proportional to the logarithm of PM concentration in the range of 0.04-400 ng mL(-1), with a detection limit of 0.018 ng mL(-1). Furthermore, the simple and convenient method had been used for PM detection in environmental and agricultural samples with satisfactory results. PMID:26143468

  2. Fluorescence tomography of biological tissue based on ultrasound tagging technique

    NASA Astrophysics Data System (ADS)

    Kobayashi, Masaki; Mizumoto, Takashi; Duc, Trinh Quang; Takeda, Motohiro

    2007-07-01

    We report a study for the development of tomographic imaging technique of fluorescence in biological tissue for assays of biological function. Ultrasonic modulation of light based on the acousto-optic effect (so called ultrasound 'tagging') is applied for imaging of fluorescence distribution in the light-scattering media. Sound-field characteristics that affect the light by modulating its amplitude through variation of the refractive index in the medium were determined. With using focused ultrasound, selectively modulated fluorescence on a depth-axis of the medium can be detected. Ultrasound tagging technique applied measuring the optical absorption in light scattering media is well known, and it is principally based on the modulation of speckle pattern. On the contrary, in the case of fluorescence, displacement of scattering particles and variation of the refractive index that is induced by density distribution in a sound field might produce the intensity modulation of scattered light. We have experimentally shown that ultrasound tagging technique is also available for fluorescence measurement. In this paper, we demonstrate the result of tomographic images of fluorescence in dense scattering media using porcine muscle as a biological tissue, and bovine adipose. Tissue samples had the dimension of 40 x 40 mm in section and fluorophore which had the 3mm size was embedded in the center of the tissue. The localized image of the fluorophore was determined with the spatial resolution of focus size of the ultrasound, suggesting the applicability of this technique for visualization of fluorescent probes in deep portion of living body.

  3. Towards a spectrum-based bar code for identification of weakly fluorescent microparticles

    NASA Astrophysics Data System (ADS)

    Petrek, Zden?k; Wiedemann, Jens; Schwille, Petra

    2014-03-01

    Spectrally resolved detection of fluorescent probes can be used to identify multiple labeled target molecules in an unknown mixture. We study how the spectral shape, the experimental noise, and the number of spectral detection channels affect the success of identification of weakly fluorescent beads on basis of their emission spectra. The proposed formalism allows to estimate the performance of the spectral identification procedure with a given set of spectral codes on the basis of the reference spectra only. We constructed a simple prism-based setup for spectral detection and demonstrate that seven distinct but overlapping spectral codes realized by combining up to three fluorescent dyes bound to a single bead in a barcode-based manner can be reliably identified. The procedure allows correct identification even in the presence of known autofluorescence background stronger than the actual signal.

  4. Innovative molecular-based fluorescent nanoparticles for multicolor single particle tracking in cells

    NASA Astrophysics Data System (ADS)

    Daniel, Jonathan; Godin, Antoine G.; Palayret, Matthieu; Lounis, Brahim; Cognet, Laurent; Blanchard-Desce, Mireille

    2016-03-01

    Based on an original molecular-based design, we present bright and photostable fluorescent organic nanoparticles (FONs) showing excellent colloidal stability in various aqueous environments. Complementary near-infrared emitting and green emitting FONs were prepared using a simple, fast and robust protocol. Both types of FONs could be simultaneously imaged at the single-particle level in solution as well as in biological environments using a monochromatic excitation and a dual-color fluorescence microscope. No evidence of acute cytotoxicity was found upon incubation of live cells with mixed solutions of FONs, and both types of nanoparticles were found internalized in the cells where their motion could be simultaneously tracked at video-rate up to minutes. These fluorescent organic nanoparticles open a novel non-toxic alternative to existing nanoparticles for imaging biological structures, compatible with live-cell experiments and specially fitted for multicolor single particle tracking.

  5. Fluorescent protein-based redox probes.

    PubMed

    Meyer, Andreas J; Dick, Tobias P

    2010-09-01

    Redox biochemistry is increasingly recognized as an integral component of cellular signal processing and cell fate decision making. Unfortunately, our capabilities to observe and measure clearly defined redox processes in the natural context of living cells, tissues, or organisms are woefully limited. The most advanced and promising tools for specific, quantitative, dynamic and compartment-specific observations are genetically encoded redox probes derived from green fluorescent protein (GFP). Within only few years from their initial introduction, redox-sensitive yellow FP (rxYFP), redox-sensitive GFPs (roGFPs), and HyPer have generated enormous interest in applying these novel tools to monitor dynamic redox changes in vivo. As genetically encoded probes, these biosensors can be specifically targeted to different subcellular locations. A critical advantage of roGFPs and HyPer is their ratiometric fluorogenic behavior. Moreover, the probe scaffold of redox-sensitive fluorescent proteins (rxYFP and roGFPs) is amenable to molecular engineering, offering fascinating prospects for further developments. In particular, the engineering of redox relays between roGFPs and redox enzymes allows control of probe specificity and enhancement of sensitivity. Genetically encoded redox probes enable the functional analysis of individual proteins in cellular redox homeostasis. In addition, redox biosensor transgenic model organisms offer extended opportunities for dynamic in vivo imaging of redox processes. PMID:20088706

  6. Fluorescence quenching of Rhodamine B base by two amines

    NASA Astrophysics Data System (ADS)

    Bakkialakshmi, S.; Selvarani, P.; Chenthamarai, S.

    2013-03-01

    Fluorescence quenching of Rhodamine B base (RhB) in DMF solution has been studied at different concentrations of the amine Triethyl amine (TEA) and n-butyl amine (NBA) at room temperature. It has been observed that the fluorescence intensity of RhB decrease with increase in the concentration of the TEA and NBA. It has been observed that the quenching due to amines proceeds via dynamic quenching process. The rate constants for the quenching process have been calculated using Stern-Volmer equation. Time resolved fluorescence study and 1H NMR spectral study have also been carried out and discussed.

  7. Tryptophan-based chromophore in fluorescent proteins can be anionic

    PubMed Central

    Sarkisyan, Karen S.; Yampolsky, Ilia V.; Solntsev, Kyril M.; Lukyanov, Sergey A.; Lukyanov, Konstantin A.; Mishin, Alexander S.

    2012-01-01

    Cyan fluorescent proteins (CFP) with tryptophan66-based chromophore are widely used for live cell imaging. In contrast to green and red fluorescent proteins, no charged states of the CFP chromophore have been described. Here, we studied synthetic CFP chromophore and found that its indole group can be deprotonated rather easily (pKa 12.4).We then reproduced this effect in the CFP mCerulean by placing basic amino acids in the chromophore microenvironment. As a result, green-emitting variant with an anionic chromophore and key substitution Val61Lys was obtained. This is the first evidence strongly suggesting that tryptophan-based chromophores in fluorescent proteins can exist in an anionic charged state. Switching between protonated and deprotonated Trp66 in fluorescent proteins represents a new unexplored way to control their spectral properties. PMID:22934131

  8. Non-contact Fluorescence-based Thermometry

    NASA Astrophysics Data System (ADS)

    Allison, Stephen W.; Gillies, George T.

    1997-03-01

    There are an increasing number of applications which are being addressed utilizing solid-state fluorescence to measure temperature. This spectroscopic approach exploits the competition between radiative and nonradiative processes for relaxation from an excited electronic state as well as thermalization and other physical mechanisms. Any laser or phosphor material can be used in this way over a characteristic temperature range. For example, oxides and aluminates activated with either rare earths or metal dopants are particularly good for high temperatures. The method is applicable from a few K to greater than 1900 K. The history of this approach is reviewed including a survey of materials, instrumentation designs and applications. Examples involve centrifuges, high speed motors and turbine engines. It is especially useful for moving surfaces and high-temperature environments, and optically hazy situations. The potential for mK resolution between the copper and aluminum triple points is excellent.

  9. Radioiodine detector based on laser induced fluorescence

    DOEpatents

    McDonald, Jimmie R. (Upper Marlboro, MD); Baronavski, Andrew P. (Alexandria, VA)

    1980-01-01

    The invention involves the measurement of the concentration of the radioisotope .sup.129 I.sub.2 in the presence of a gas. The invention uses a laser to excite a sample of the .sup.129 I.sub.2 in a sample gas chamber and a reference sample of a known concentration of .sup.129 I.sub.2 in a reference gas chamber. The .sup.129 I.sub.2 in the sample and reference gas chamber each gives off fluorescence emissions which are received by photomultipliers which provide signals to a detector. The detector uses a ratioing technique to determine the concentration of .sup.129 I.sub.2 in the sample gas chamber.

  10. Development of fluorescent lead II sensor based on an anthracene derived chalcone

    NASA Astrophysics Data System (ADS)

    Prabhu, J.; Velmurugan, K.; Nandhakumar, R.

    2015-06-01

    A simple anthracene based chalcone as a fluorescent chemosensor 1, capable of detecting Pb2+ in aqueous media, has been synthesized by the reaction between pyridine 2-carboxaldehyde and 9-acetyl anthracene. The Pb2+ recognition processes follows a photo induced electron transfer (PET) mechanism and are scarcely influenced by other coexisting metal ions. In addition, determination of lead in a variety of samples was also determined.

  11. Development of fluorescent lead II sensor based on an anthracene derived chalcone.

    PubMed

    Prabhu, J; Velmurugan, K; Nandhakumar, R

    2015-06-01

    A simple anthracene based chalcone as a fluorescent chemosensor 1, capable of detecting Pb(2+) in aqueous media, has been synthesized by the reaction between pyridine 2-carboxaldehyde and 9-acetyl anthracene. The Pb(2+) recognition processes follows a photo induced electron transfer (PET) mechanism and are scarcely influenced by other coexisting metal ions. In addition, determination of lead in a variety of samples was also determined. PMID:25744532

  12. Development of fluorescent lead II sensor based on an anthracene derived chalcone.

    TOXLINE Toxicology Bibliographic Information

    Prabhu J; Velmurugan K; Nandhakumar R

    2015-06-05

    A simple anthracene based chalcone as a fluorescent chemosensor 1, capable of detecting Pb(2+) in aqueous media, has been synthesized by the reaction between pyridine 2-carboxaldehyde and 9-acetyl anthracene. The Pb(2+) recognition processes follows a photo induced electron transfer (PET) mechanism and are scarcely influenced by other coexisting metal ions. In addition, determination of lead in a variety of samples was also determined.

  13. A Simple Visualization of Double Bond Properties: Chemical Reactivity and UV Fluorescence

    ERIC Educational Resources Information Center

    Grayson, Scott M.

    2012-01-01

    A simple, easily visualized thin-layer chromatography (TLC) staining experiment is presented that highlights the difference in reactivity between aromatic double bonds and nonaromatic double bonds. Although the stability of aromatic systems is a major theme in organic chemistry, the concept is rarely reinforced "visually" in the undergraduate…

  14. A Simple Visualization of Double Bond Properties: Chemical Reactivity and UV Fluorescence

    ERIC Educational Resources Information Center

    Grayson, Scott M.

    2012-01-01

    A simple, easily visualized thin-layer chromatography (TLC) staining experiment is presented that highlights the difference in reactivity between aromatic double bonds and nonaromatic double bonds. Although the stability of aromatic systems is a major theme in organic chemistry, the concept is rarely reinforced "visually" in the undergraduate

  15. Field portable mobile phone based fluorescence microscopy for detection of Giardia lamblia cysts in water samples

    NASA Astrophysics Data System (ADS)

    Ceylan Koydemir, Hatice; Gorocs, Zoltan; McLeod, Euan; Tseng, Derek; Ozcan, Aydogan

    2015-03-01

    Giardia lamblia is a waterborne parasite that causes an intestinal infection, known as giardiasis, and it is found not only in countries with inadequate sanitation and unsafe water but also streams and lakes of developed countries. Simple, sensitive, and rapid detection of this pathogen is important for monitoring of drinking water. Here we present a cost-effective and field portable mobile-phone based fluorescence microscopy platform designed for automated detection of Giardia lamblia cysts in large volume water samples (i.e., 10 ml) to be used in low-resource field settings. This fluorescence microscope is integrated with a disposable water-sampling cassette, which is based on a flow-through porous polycarbonate membrane and provides a wide surface area for fluorescence imaging and enumeration of the captured Giardia cysts on the membrane. Water sample of interest, containing fluorescently labeled Giardia cysts, is introduced into the absorbent pads that are in contact with the membrane in the cassette by capillary action, which eliminates the need for electrically driven flow for sample processing. Our fluorescence microscope weighs ~170 grams in total and has all the components of a regular microscope, capable of detecting individual fluorescently labeled cysts under light-emitting-diode (LED) based excitation. Including all the sample preparation, labeling and imaging steps, the entire measurement takes less than one hour for a sample volume of 10 ml. This mobile phone based compact and cost-effective fluorescent imaging platform together with its machine learning based cyst counting interface is easy to use and can even work in resource limited and field settings for spatio-temporal monitoring of water quality.

  16. Wide field-of-view Talbot grid-based microscopy for multicolor fluorescence imaging.

    PubMed

    Pang, Shuo; Han, Chao; Erath, Jessey; Rodriguez, Ana; Yang, Changhuei

    2013-06-17

    The capability to perform multicolor, wide field-of-view (FOV) fluorescence microscopy imaging is important in screening and pathology applications. We developed a microscopic slide-imaging system that can achieve multicolor, wide FOV, fluorescence imaging based on the Talbot effect. In this system, a light-spot grid generated by the Talbot effect illuminates the sample. By tilting the excitation beam, the Talbot-focused spot scans across the sample. The images are reconstructed by collecting the fluorescence emissions that correspond to each focused spot with a relay optics arrangement. The prototype system achieved an FOV of 12 10 mm(2) at an acquisition time as fast as 23 s for one fluorescence channel. The resolution is fundamentally limited by spot size, with a demonstrated full-width at half-maximum spot diameter of 1.2 ?m. The prototype was used to nimage green fluorescent beads, double-stained human breast cancer SK-BR-3 cells, Giardia lamblia cysts, and the Cryptosporidium parvum oocysts. This imaging method is scalable and simple for implementation of high-speed wide FOV fluorescence microscopy. PMID:23787643

  17. Wide field-of-view Talbot grid-based microscopy for multicolor fluorescence imaging

    PubMed Central

    Pang, Shuo; Han, Chao; Erath, Jessey; Rodriguez, Ana; Yang, Changhuei

    2013-01-01

    The capability to perform multicolor, wide field-of-view (FOV) fluorescence microscopy imaging is important in screening and pathology applications. We developed a microscopic slide-imaging system that can achieve multicolor, wide FOV, fluorescence imaging based on the Talbot effect. In this system, a light-spot grid generated by the Talbot effect illuminates the sample. By tilting the excitation beam, the Talbot-focused spot scans across the sample. The images are reconstructed by collecting the fluorescence emissions that correspond to each focused spot with a relay optics arrangement. The prototype system achieved an FOV of 12 10 mm2 at an acquisition time as fast as 23 s for one fluorescence channel. The resolution is fundamentally limited by spot size, with a demonstrated full-width at half-maximum spot diameter of 1.2 ?m. The prototype was used to image green fluorescent beads, double-stained human breast cancer SK-BR-3 cells, Giardia lamblia cysts, and the Cryptosporidium parvum oocysts. This imaging method is scalable and simple for implementation of high-speed wide FOV fluorescence microscopy. PMID:23787643

  18. A Simple Inquiry-Based Lab for Teaching Osmosis

    ERIC Educational Resources Information Center

    Taylor, John R.

    2014-01-01

    This simple inquiry-based lab was designed to teach the principle of osmosis while also providing an experience for students to use the skills and practices commonly found in science. Students first design their own experiment using very basic equipment and supplies, which generally results in mixed, but mostly poor, outcomes. Classroom "talk…

  19. A Simple Inquiry-Based Lab for Teaching Osmosis

    ERIC Educational Resources Information Center

    Taylor, John R.

    2014-01-01

    This simple inquiry-based lab was designed to teach the principle of osmosis while also providing an experience for students to use the skills and practices commonly found in science. Students first design their own experiment using very basic equipment and supplies, which generally results in mixed, but mostly poor, outcomes. Classroom "talk

  20. Motor Oil Classification Based on Time-Resolved Fluorescence

    PubMed Central

    Mu, Taotao; Chen, Siying; Zhang, Yinchao; Guo, Pan; Chen, He; Meng, Fandong

    2014-01-01

    A time-resolved fluorescence (TRF) technique is presented for classifying motor oils. The system is constructed with a third harmonic Nd:YAG laser, a spectrometer, and an intensified charge coupled device (ICCD) camera. Steady-state and time-resolved fluorescence (TRF) measurements are reported for several motor oils. It is found that steady-state fluorescence is insufficient to distinguish the motor oil samples. Then contour diagrams of TRF intensities (CDTRFIs) are acquired to serve as unique fingerprints to identify motor oils by using the distinct TRF of motor oils. CDTRFIs are preferable to steady-state fluorescence spectra for classifying different motor oils, making CDTRFIs a particularly choice for the development of fluorescence-based methods for the discrimination and characterization of motor oils. The two-dimensional fluorescence contour diagrams contain more information, not only the changing shapes of the LIF spectra but also the relative intensity. The results indicate that motor oils can be differentiated based on the new proposed method, which provides reliable methods for analyzing and classifying motor oils. PMID:24988439

  1. A New Type Of Fiber Optic Biosensor Based On The Intrinsic Fluorescence Of Immobilized Flavoproteins

    NASA Astrophysics Data System (ADS)

    Wolfbeis, Otto S.; Trettnak, Wolfgang

    1990-02-01

    We describe a new biosensor for monitoring the concentration of enzyme glucose, lactate, and other substrates that are metabolized by an oxidation process. The method is based on the finding that enzymes having FAD as a prosthetic group change their fluorescence during interaction with a substrate. Typical enzymes that have been studied include glucose oxidase (GOD), lactate mono-oxygenase (LMO), and cholesterol oxidase (ChOD). Their fluorescence is monitored via fiber optic light guides at wavelengths above 500 nm, following fluorescence excitation at around 410 - 450 nm. The relative fluorescence intensities of the enzymes vary to a large extent, being highest for LMO, and rather low for ChOD. Typical detection limits are in the 0.5 mM range for lactate and 1.5 mM for glucose at ambient oxygen pressure. A characteristic feature of this sensor is the narrow dynamic range which usually does not exceed 3 mM. This can be explained in terms of enzyme kinetics and diffusional processes. Unlike optical biosensors based on measurement of the intrinsic fluorescence of NADH, this sensor type has the advantages of full reversibility (because reduced FAD-based enzymes accept oxygen as a second substrate) and analytical wavelengths that are compatible with plastic or glass fiber optics. It is fairly simple in construction because the enzyme acts as both the recognition and transduction element. The method also has been applied successfully in an flow injection analysis-like type of arrangement.

  2. Selective fluorescence detection of monosaccharides using a material composite formed between graphene oxide and boronate-based receptors.

    PubMed

    Sun, Xiaolong; Zhu, Bin; Ji, Ding-Kun; Chen, Qibin; He, Xiao-Peng; Chen, Guo-Rong; James, Tony D

    2014-07-01

    We have developed a novel class of simple materials for sensing monosaccharides by the functionalization of graphene oxide (GO) with boronate-based fluorescence probes (BA1 and BA2). The composite materials were characterized by atomic force microscopy, Raman spectroscopy, and UV-vis/fluorescence spectroscopy. The strong fluorescence of the BA probes is quenched in the presence of GO through fluorescence resonance energy transfer. The BA@GO composite sensors formed provide a useful platform for fluorogenic detection of monosaccharides based on the strong affinity between the boronic acid receptor and monosaccharides. The BA@GO composite sensor displayed a "turn-on" fluorescence response with a good linear relationship toward fructose over a range of other saccharides. PMID:24918717

  3. Simple refractometer based on in-line fiber interferometers

    NASA Astrophysics Data System (ADS)

    Esteban, .; Martnez Manuel, R.; Shlyagin, M. G.

    2015-09-01

    A very simple but accurate optical fiber refractometer based on the Fresnel reflection in the fiber tip and two in-line low-reflective mirrors for light intensity referencing is reported. Each mirror was generated by connecting together 2 fiber sections with FC/PC and FC/APC connectors using the standard FC/PC mating sleeve. For the sensor interrogation, a standard DFB diode laser pumped with a sawtooth-wave current was used. A resolution of 6 x 10-4 was experimentally demonstrated using different liquids. A simple sensor construction and the use of low cost components make the reported system interesting for many applications.

  4. Transparency-based microplates for fluorescence quantification.

    PubMed

    Cheong, Brandon Huey-Ping; Diep, Vu; Ng, Tuck Wah; Liew, Oi Wah

    2012-03-01

    Microplates for use in resource-limited laboratories should ideally not require processes that involve substantial large-scale production in order to be viable. We describe and demonstrate here an approach of using a silicone sheet with holes, conveniently cut out precisely using an inexpensive cutting plotter to correspond with regions where liquid is to be dispensed, and attaching it to a transparency to create very thin well arrays. With this, the contact angle hysteresis behavior of liquid could be harnessed to produce taller drop shapes so that the fiber probe used could read in the emitted light more effectively. Experimentation conducted revealed fluorescence measurements that were significantly more sensitive than standard microplates, notwithstanding that smaller volumes of liquid were needed. This was achieved using both the fiber optic and imaging evaluation modes. The two methods investigated, one with a lid placed and one without, showed the latter to produce marginally more sensitive readings as opposed to improved immunity from the environment with the former. These favorable measurement characteristics were found to be achievable with an estimated production cost of AU $0.40 and fabrication times of 3.5 min (96 wells) and 6.5 min (384 wells) per plate. PMID:22266206

  5. Simple, rapid detection of influenza A (H1N1) viruses using a highly sensitive peptide-based molecular beacon.

    PubMed

    Lim, Eun-Kyung; Guk, Kyeonghye; Kim, Hyeran; Chung, Bong-Hyun; Jung, Juyeon

    2015-12-15

    A peptide-based molecular beacon (PEP-MB) was prepared for the simple, rapid, and specific detection of H1N1 viruses using a fluorescence resonance energy transfer (FRET) system. The PEP-MB exhibited minimal fluorescence in its "closed" hairpin structure. However, in the presence of H1N1 viruses, the specific recognition of the hemagglutinin (HA) protein of H1 strains by the PEP-MB causes the beacon to assume an "open" structure that emits strong fluorescence. The PEP-MB could detect H1N1 viruses within 15 min or even 5 min and can exhibit strong fluorescence even at low viral concentrations, with a detection limit of 4 copies. PMID:26509476

  6. A chelation enhanced selective fluorescence sensing of Hg2+ by a simple quinoline substituted tripodal amide receptor.

    PubMed

    Ahamed, B Nisar; Ghosh, Pradyut

    2011-12-14

    Two tris(2-aminoethyl)amine (tren) based tripodal amide fluoroionophores, 1 and 2, functionalized with quinoline (chelating fluorophore) and naphthalene (non-chelating fluorophore) respectively, are synthesized in good yields. Fluoroionophore 1 shows a selective UV-Vis spectral shift in the case of Hg(2+) in acetonitrile among different metal ions like Li(+), Na(+), Ca(2+), Mg(2+), Cr(2+), Mn(2+), Fe(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), Hg(2+), Pb(2+), and Ag(+). On the other hand, fluoroionophore 2 shows no selectivity towards any of the above metal ions in the UV-Vis study. Furthermore, 1 shows a selective chelation induced fluorescence enhancement in the presence of Hg(2+) whereas 2 shows the enhancement of fluorescence with most of the metal ions via a photoinduced charge transfer mechanism. The naked eye detection of Hg(2+) in an acetonitrile solution of 1 shows a greenish fluorescence upon UV light irradiation. The isolated Hg(2+) complex of 1, 3, shows a similar UV-Vis and fluorescence spectral output as observed from in situ spectroscopic studies of 1 in the presence of Hg(2+). Infra-red (IR) and (1)H- NMR studies also reveal the interaction of Hg(2+) with the quinoline nitrogen atoms as well as with the amide functionality. PMID:21984526

  7. mb-FLIM: model-based fluorescence lifetime imaging

    NASA Astrophysics Data System (ADS)

    Zhao, Qiaole; Young, Ian Ted; Schouten, Raymond; Stallinga, Sjoerd; Jalink, Kees; de Jong, Sander

    2012-03-01

    We have developed a model-based, parallel procedure to estimate fluorescence lifetimes. Multiple frequencies are present in the excitation signal. Modeling the entire fluorescence and measurement process produces an analytical ratio of polynomials in the lifetime variable ?. A non-linear model-fitting procedure is then used to estimate ?. We have analyzed this model-based approach by simulating a 10 ?M fluorescein solution (? = 4 ns) and all relevant noise sources. We have used real LED data to drive the simulation. Using 240 ?s of data, we estimate ? = 3.99 ns. Preliminary experiments on real fluorescent images taken from fluorescein solutions (measured ? = 4.1 ns), green plastic test slides (measured ? = 3.0 ns), and GFP in U2OS (osteosarcoma) cells (measured ? = 2.1 ns) demonstrate that this model-based measurement technique works.

  8. A fluorescence-based assay for nanogram quantification of proteins using a protein binding ligand.

    PubMed

    Shrivastava, H Yamini; Nair, Balachandran Unni

    2003-01-01

    Fluorescence emission has been investigated in the context of estimation of proteins at nanogram levels. A Schiff base ligand with donor-acceptor substituents has been utilized as a fluorescent probe. The potency of this ligand is that it possesses the binding sites for both hydrophobic as well as hydrophilic groups in the proteins. The fluorescence emission of the probe was enhanced in the presence of nanogram levels of protein, which clearly signifies that even the least concentration of the protein is sufficient to perturb the environment around the probe. We demonstrate here that the fluorescence characteristic of the probe can be utilized to estimate even nanogram levels (66 ng-1 microgram mL(-1)) of protein. The major limitation of the currently available standard methods is the range of protein estimation, which terminates at microgram level and the interference due to the specificity of the amino acids, which vary from proteins to proteins. This fluorescence emission-based method is free from interference from any type of buffers, ionic strength of the medium and any specific amino acid residue and is a simple, rapid, single-step, sensitive method of estimation which can be applied to different classes of proteins. PMID:12520454

  9. Picosecond fluorescence of simple photosynthetic membranes: Evidence of spectral inhomogeneity and directed energy transfer

    NASA Astrophysics Data System (ADS)

    Pullerits, T?nu; Freiberg, Arvi

    1991-01-01

    The picosecond time-domain singlet excitation transfer and trapping kinetics in photosynthetic membranes in case of low excitation intensities is studied by numerical integration of the appropriate master equation. The essential features of our two-dimensional-lattice random walk model are spectral heterogeneity of the light-harvesting antenna, inclusion of temperature effects, nonabsolute excitation trap, correlation between spectral and spatial parameters. A reasonably good agreement between theoretical and experimental fluorescence decay kinetics for purple photosynthetic bacterium Rhodospirillum rubrum is achieved only by assuming relatively large spectral inhomogeneity. From this comparison the average excitation lifetime on the lattice site is estimated to be 5-8 ps at the effective nearest neighbour lattice distance of 32 . If the model is correct, the relatively slow hopping rate determines that excitation transfer and trapping in R. rubrum at active photosynthesis conditions is a diffusion-limited process. The invariably present spectral disorder of photosynthetic systems promoting directed energy transfer serves for higher light-utilizing efficiency.

  10. A simple, rapid and low-cost staining method for gel-electrophoresis separated phosphoproteins via the fluorescent purpurin dye.

    PubMed

    Cong, Weitao; Shen, Jiayi; Xuan, Yuanhu; Zhu, Xinliang; Ni, Maowei; Zhu, Zhongxin; Hong, Guoying; Lu, Xianghong; Jin, Litai

    2014-12-01

    A novel fluorescence detection method for phosphoproteins in 1-D and 2-D SDS-PAGE by using purpurin is developed in this study. Phosphoproteins as low as 4-8 ng could be specifically detected by purpurin within 60 min, and the detection limit is similar to or better than that of Pro-Q Diamond staining. Only 2 steps (staining and destaining) are needed for purpurin staining without requiring excessive fixing and washing steps, and for single use, $0.8 is enough for purpurin staining. By comprehensively comparing with Pro-Q Diamond staining, it is concluded that purpurin staining is a simple, rapid and low-cost staining method for a broad application to the research of phosphoproteins. PMID:25325196

  11. A simple turn on fluorescent sensor for the selective detection of thiamine using coconut water derived luminescent carbon dots.

    PubMed

    Purbia, Rahul; Paria, Santanu

    2016-05-15

    In this study microwave-assisted hydrothermal method was used to prepare highly luminescent carbon dots (1-6nm size) within a minute from tender coconut (Cocos nucifera) water. The synthesized carbon dots (C-dots) exhibit emission of blue and green lights while excited at 390 and 450nm wavelengths, respectively. As an application, these C-dots were tested for a simple "turn on" fluorescent sensor for rapid detection of thiamine (vitamin B1). The detection of thiamine in human body is very important to prevent various diseases such as beriberi, neurological disorders, optic neuropathy, etc. The fluorescence emission intensity of C-dots quenches after addition of Cu(2+) ion and then again increases selectively (turn on) after the addition of thiamine. The fluorescence emission intensity enhancement of Cu(2+) ion modified C-dots in the presence of thiamine exhibits a linear relationship within the thiamine concentration range of 10-50μM. The limit of detection was found to be 280nM from this study. The selectivity of the detection was also tested in the presence of different organic molecules and inorganic ions (Ca(2+), Mg(2+), Na(+), K(+), Cl(-), SO4(2-), and NO3(-)) which are present in blood serum and urine and found to be almost no interference in the detection. Finally, to see the applicability in real samples a commercial vitamin capsule was tested and found less than 3% error in the detected concentration. The C-dots were also used for bioimaging of fungus and the results show they are also suitable for this application too. PMID:26745793

  12. A simple radioisotope X-ray fluorescence method for provenance studies of archaeological ceramics employing principal component analysis

    NASA Astrophysics Data System (ADS)

    LaBrecque, J. J.; Vaz, J. E.; Cruxent, J. M.; Rosales, P. A.

    1998-01-01

    A simple radioisotope X-ray fluorescence method of 1000 s irradiation of the samples by a 109Cd source combined with principal component analysis is described for determining the relative mass fractions of trace elements in majolica ceramics for provenance classification. Six provenances from Europe using 29 samples as standards and 12 unknown samples were investigated and characterized using selected trace elements as the variables. The unknown samples were previously assigned, but not definitively, by stylistic analysis and/or thermoluminescence measurements to the provenances of Teruel (Spain) and Holland. Because of a moderate fluorescence time, only the four net peak intensities of Pb, Rb, Sr and Zr could be used as variables. We have also studied the effect of not including the Pb variable, since the clay matrixes could have been contaminated in the glazing process or when the Pb-Sn enamel was removed. It is shown in both cases that the results were more consistent with the stylistic analysis and thermoluminescence measurements when the Pb concentration variable was not considered. A comparison of principal component analysis employing the three elements was similar to plotting of the relative mass fractions on a triangle graph.

  13. Highly sensitive fluorescence and SERS detection of azide through a simple click reaction of 8-chloroquinoline and phenylacetylene.

    PubMed

    Zeng, Qing; Ye, Lingling; Ma, Lu; Yin, Wenqing; Li, Tingsheng; Liang, Aihui; Jiang, Zhiliang

    2015-05-01

    In 0.19 mol/L acetic acid (HAc), a click reaction of 8-chloroquinoline/azide/phenylacetylene take places in aqueous solution without Cu(I) as a catalyst. 8-Chloroquinoline (CQN) exhibited a strong fluorescence peak at 430 nm that was quenched linearly as the concentration of azide increased from 20 to 1000 ng/mL. This quenching was due to consumption of CQN in the click reaction and a decrease in the number of efficiently excited photons due to the presence of triazole-quinoline ramification molecules with strong hydrophobicity. Using blue nanosilver sol as the substrate, CQN absorbed onto the surface of nanosilver particles, showing a strong surface-enhanced Raman scattering (SERS) peak at 1585 cm(-1) that decreased linearly as the azide concentration increased from 8 to 500 ng/mL; the detection limit was 4 ng/mL. Thus, two new, simple and sensitive fluorescence and SERS methods have been developed for the determination of azide via the click reaction. PMID:25045121

  14. Fluorescence lifetime-based glucose sensor using NADH

    NASA Astrophysics Data System (ADS)

    von Ketteler, A.; Siegberg, D.; Herten, D. P.; Horn, C.; Petrich, W.

    2012-03-01

    Fluorescence lifetime-based glucose sensing does not depend on fluctuations of the intensity of the light source, light scattering, or changes in the transmission of optical components. Here we demonstrate the sensing of glucose based on the fluorescence lifetime properties of dihydro nicotinamide adenine dinucleotide (NADH), which is reduced from NAD in the presence of glucose and glucose dehydrogenase. In particular we use the difference in the fluorescence properties of free and protein-bound NADH and calculate an average fluorescence lifetime, which arises from the two short lifetimes ?1=0.28ns and ?2=0.60ns (representing free NADH) and the longer lifetime of ?3=2.9ns (for the protein-bound NADH). While initial results were derived from measurements in aqueous solution, we also demonstrate the suitability of this method for determining the concentration of glucose in blood using test strips. We find that the average fluorescence lifetime changes linearly by a factor of 0.17 per 100mg/dl change in glucose concentration. As an alternative the ratio between free and protein-bound components Rs/l may also be used for quantification. Rs/l increases by a factor of 0.74 per 100mg/dl change in glucose concentration.

  15. Direct detection of sulfide ions [S2-] in aqueous media based on fluorescence quenching of functionalized CdS QDs at trace levels: analytical applications to environmental analysis.

    PubMed

    Gore, Anil H; Vatre, Sandip B; Anbhule, Prashant V; Han, Sung-Hwan; Patil, Shivajirao R; Kolekar, Govind B

    2013-03-01

    A novel, simple but highly selective fluorescent probe is developed for the direct detection of sulfide ions [S(2-)] based on the fluorescence quenching of the functionalized CdS QDs in aqueous solution at trace levels and successfully applied for quantitation of S(2-) from water samples in a complex matrix exclusive of pretreatment by standard addition method. PMID:23334151

  16. A graphene oxide-based platform for the assay of RNA synthesis by RNA polymerase using a fluorescent peptide nucleic acid probe.

    PubMed

    Park, Joon Soo; Baek, Ahruem; Park, Il-Soo; Jun, Bong-Hyun; Kim, Dong-Eun

    2013-10-14

    We report a simple, direct fluorometric assay based on graphene oxide (GO) for RNA polymerase-mediated RNA synthesis. In principle, fluorescent peptide nucleic acid (PNA) probes were designed, and annealed with RNA products and the resultant RNA-PNA hybrids induced the recovery of fluorescence intensity of the PNA probes adsorbed onto the GO surface. PMID:23995852

  17. Wireless implantable electronic platform for chronic fluorescent-based biosensors.

    PubMed

    Valdastri, Pietro; Susilo, Ekawahyu; Förster, Thilo; Strohhöfer, Christof; Menciassi, Arianna; Dario, Paolo

    2011-06-01

    The development of a long-term wireless implantable biosensor based on fluorescence intensity measurement poses a number of technical challenges, ranging from biocompatibility to sensor stability over time. One of these challenges is the design of a power efficient and miniaturized electronics, enabling the biosensor to move from bench testing to long term validation, up to its final application in human beings. In this spirit, we present a wireless programmable electronic platform for implantable chronic monitoring of fluorescent-based autonomous biosensors. This system is able to achieve extremely low power operation with bidirectional telemetry, based on the IEEE802.15.4-2003 protocol, thus enabling over three-year battery lifetime and wireless networking of multiple sensors. During the performance of single fluorescent-based sensor measurements, the circuit drives a laser diode, for sensor excitation, and acquires the amplified signals from four different photodetectors. In vitro functionality was preliminarily tested for both glucose and calcium monitoring, simply by changing the analyte-binding protein of the biosensor. Electronics performance was assessed in terms of timing, power consumption, tissue exposure to electromagnetic fields, and in vivo wireless connectivity. The final goal of the presented platform is to be integrated in a complete system for blood glucose level monitoring that may be implanted for at least one year under the skin of diabetic patients. Results reported in this paper may be applied to a wide variety of biosensors based on fluorescence intensity measurement. PMID:21385666

  18. Simple Method to Enhance the Photostability of the Fluorescence Reporter R6G for Prolonged Single Molecule Studies

    PubMed Central

    Guo, Lin; Gai, Feng

    2013-01-01

    For fluorescence-based single-molecule studies, photobleaching of the dye reporter often limits the time window over which individual molecules can be followed. As such, many strategies, for example using a cocktail of chemical reagents, have been developed to decrease the rate of photobleaching. Herein, we introduce a new and highly effective method to enhance the photostability of one of the commonly used fluorescent dyes, rhodamine 6G (R6G). We show that micron-sized polydimethylsiloxane (PDMS) wells, when the PDMS surface is properly treated, not only provide a confined environment for single-molecule detection, but can also significantly increase the survival time of individual R6G molecules before photobleaching. Moreover, our results suggest, consistent with several previous studies, that R6G photobleaching involves a radical state. PMID:23641719

  19. Graphene oxide based fluorescent aptasensor for adenosine deaminase detection using adenosine as the substrate.

    PubMed

    Xing, Xiao-Jing; Liu, Xue-Guo; Yue-He; Luo, Qing-Ying; Tang, Hong-Wu; Pang, Dai-Wen

    2012-01-01

    We present a novel fluorescent aptasensor for simple and accurate detection of adenosine deaminase (ADA) activity and inhibition on the basis of graphene oxide (GO) using adenosine (AD) as the substrate. This aptasensor consists of a dye-labeled single-stranded AD specific aptamer, GO and AD. The fluorescence intensity of the dye-labeled AD specific aptamer is quenched very efficiently by GO as a result of strong ?-? stacking interaction and excellent electronic transference of GO. In the presence of AD, the fluorescence of the GO-based probe is recovered since the competitive binding of AD and GO with the dye-labeled aptamer prevents the adsorption of dye-labeled aptamer on GO. When ADA was introduced to this GO-based probe solution, the fluorescence of the probe was quenched owing to ADA can convert AD into inosine which has no affinity to the dye-labeled aptamer, thus allowing quantitative investigation of ADA activity. The as-proposed sensor is highly selective and sensitive for the assay of ADA activity with a detection limit of 0.0129U/mL in clean buffer, which is more than one order of magnitude lower than the previous reports. Meanwhile, a good linear relationship with the correlation coefficient of R=0.9922 was obtained by testing 5% human serum containing a series of concentrations of ADA. Additionally, the inhibition effect of erythro-9-(2-hydroxy-3-nonyl) adenine on ADA activity was investigated in this design. The GO-based fluorescence aptasensor not only provides a simple, cost-effective and sensitive platform for the detection of ADA and its inhibitor but also shows great potential in the diagnosis of ADA-relevant diseases and drug development. PMID:22613226

  20. Simple method of DNA stretching on glass substrate for fluorescence image and spectroscopy

    NASA Astrophysics Data System (ADS)

    Neupane, Guru P.; Dhakal, Krishna P.; Lee, Hyunsoo; Guthold, Martin; Joseph, Vincent S.; Hong, Jong-Dal; Kim, Jeongyong

    2013-05-01

    Study of biological molecule DNA has contributed to developing many breaking thoughts and wide applications in multidisciplinary fields, such as genomic, medical, sensing and forensic fields. Stretching of DNA molecules is an important supportive tool for AFM or spectroscopic studies of DNA in a single molecular level. In this article, we established a simple method of DNA stretching (to its full length) that occurred on a rotating negatively-charged surface of glass substrate. The isolation of a single DNA molecule was attained by the two competitive forces on DNA molecules, that is, the electrostatic attraction developed between the positively charged YOYO-1 stained DNA and the negatively charged substrate, and the centrifugal force of the rotating substrate, which separates the DNA aggregates into the single molecule. Density of stretched DNA molecules was controlled by selecting the specific parameters such as spinning time and rates, loading volume of DNA-dye complex solution etc. The atomic force microscopy image exhibited a single DNA molecule on the negatively-charged substrate in an isolated state. Further, the photoluminescence spectra of a single DNA molecule stained with YOYO-1 were achieved using the method developed in the present study, which is strongly believed to effectively support the spectroscopic analysis of DNA in a single molecular level.

  1. Magnetite nanoparticles for biosensor model based on bacteria fluorescence

    NASA Astrophysics Data System (ADS)

    Poita, A.; Creanga, D.-E.; Airinei, A.; Tupu, P.; Goiceanu, C.; Avadanei, O.

    2009-06-01

    Fluorescence emission of pyoverdine - the siderophore synthesized by iron scavenger bacteria - was studied using in vitro cultures of Pseudomonas aeruginosa with the aim to design a biosensor system for liquid sample iron loading. Diluted suspensions of colloidal magnetite nanoparticles were supplied in the culture medium (10 microl/l and 100 microl/l) to simulate magnetic loading with iron oxides of either environmental waters or human body fluids. The electromagnetic exposure to radiofrequency waves of bacterial samples grown in the presence of magnetic nanoparticles was also carried out. Cell density diminution but fluorescence stimulation following 10 microl/l ferrofluid addition and simultaneous exposure to radiofrequency waves was evidenced. The inhibitory influence of 100 microl/l ferrofluid combined with RF exposure was evidenced by fluorescence data. Mathematical model was proposed to approach quantitatively the dynamics of cell density and fluorescence emission in relation with the consumption of magnetite nanoparticle supplied medium. The biosensor scheme was shaped based on the response to iron loading of bacterial sample fluorescence.

  2. Fluorescence based explosive detection: from mechanisms to sensory materials.

    PubMed

    Sun, Xiangcheng; Wang, Ying; Lei, Yu

    2015-11-21

    The detection of explosives is one of the current pressing concerns in global security. In the past few decades, a large number of emissive sensing materials have been developed for the detection of explosives in vapor, solution, and solid states through fluorescence methods. In recent years, great efforts have been devoted to develop new fluorescent materials with various sensing mechanisms for detecting explosives in order to achieve super-sensitivity, ultra-selectivity, as well as fast response time. This review article starts with a brief introduction on various sensing mechanisms for fluorescence based explosive detection, and then summarizes in an exhaustive and systematic way the state-of-the-art of fluorescent materials for explosive detection with a focus on the research in the recent 5 years. A wide range of fluorescent materials, such as conjugated polymers, small fluorophores, supramolecular systems, bio-inspired materials and aggregation induced emission-active materials, and their sensing performance and sensing mechanism are the centerpiece of this review. Finally, conclusions and future outlook are presented and discussed. PMID:26335504

  3. A Cell-Based Fluorescent Assay to Detect the Activity of Shiga Toxin and Other Toxins That Inhibit Protein Synthesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Escherichia coli O157:H7, a major cause of food-borne illness, produces Shiga toxins that block protein synthesis by inactivating the ribosome. In this chapter we describe a simple cell-based fluorescent assay to detect Shiga toxins and inhibitors of toxin activity. The assay can also be used to d...

  4. Performance validation of EMCCD and ICCD based near-infrared fluorescence imaging systems on a fluorescence solid phantom

    NASA Astrophysics Data System (ADS)

    Zhu, Banghe; Sevick-Muraca, Eva M.

    2012-03-01

    Near infrared (NIR) fluorescence imaging has been successfully applied for non-invasive assessment of both lymphatic architecture and function as well as potential disease markers of lymphatic dysfunction in clinical studies with intradermal injection of indocyanine green (ICG). For new "first-in-humans" NIR fluorescence imaging agents that need to be employed at far lower quantities, NIR fluorescence imaging devices with high measurement sensitivity are most favorable. However, the measurement sensitivity of NIR fluorescence imaging devices is limited by various parameters, including quantum efficiency of CCD chip, noise sources in the CCD camera, and the leakage of excitation light through optical filters. In this contribution, we present a quantum dot-based fluorescence solid phantom and its use for characterization of excitation light leakage and measurement sensitivity in both the intensified CCD (ICCD) and Electron Multiplying CCD (EMCCD) based NIR fluorescence imaging devices. The stability of the constructed quantum dot-based fluorescence solid phantom was first demonstrated and used to demonstrate higher measurement sensitivity compared of the ICCD as opposed to the EMCCD based NIR fluorescence imaging device when integration time were maintained less than 1.0 s. The phantom was used to assess the calculated transmission ratio, R, to minimize noise owing to excitation light leakage and show optimized filtering capabilities. The constructed quantum dot based solid phantom and the methodology for measuring parameters of transmission ratio and SNR can be used as a standard and quantifiable metric for installation and operational qualification of all NIR fluorescence imaging devices.

  5. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye.

    PubMed

    Nath, Nidhi; Godat, Becky; Zimprich, Chad; Dwight, Stephen J; Corona, Cesear; McDougall, Mark; Urh, Marjeta

    2016-04-01

    Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer antibody therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of antibody drug conjugates (ADCs), efficient removal of surface receptors from cancer cells and changing the pharmacokinetics profile of the antibody drugs are some of key ways that internalization impacts the therapeutic efficacy of the antibodies. Over the years, several techniques have been used to study antibody internalization including radiolabels, fluorescent microscopy, flow cytometry and cellular toxicity assays. While these methods allow analysis of internalization, they have limitations including a multistep process and limited throughput and are generally endpoint assays. Here, we present a new homogeneous method that enables time and concentration dependent measurements of antibody internalization. The method uses a new hydrophilic and bright pH sensor dye (pHAb dye), which is not fluorescent at neutral pH but becomes highly fluorescent at acidic pH. For receptor mediated antibody internalization studies, antibodies against receptors are conjugated with the pHAb dye and incubated with the cells expressing the receptors. Upon binding to the receptor, the dyes conjugated to the antibody are not fluorescent because of the neutral pH of the media, but upon internalization and trafficking into endosomal and lysosomal vesicles the pH drops and dyes become fluorescent. The enabling attributes of the pHAb dyes are the hydrophilic nature to minimize antibody aggregation and bright fluorescence at acidic pH which allows development of simple plate based assays using a fluorescent reader. Using two different therapeutic antibodies - Trastuzumab (anti-HER2) and Cetuximab (anti-EGFR) - we show labeling with pHAb dye using amine and thiol chemistries and impact of chemistry and dye to antibody ration on internalization. We finally present two new approaches using the pHAb dye, which will be beneficial for screening a large number of antibody samples during early monoclonal development phase. PMID:26851520

  6. Broccoli: rapid selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution.

    PubMed

    Filonov, Grigory S; Moon, Jared D; Svensen, Nina; Jaffrey, Samie R

    2014-11-19

    Genetically encoded fluorescent ribonucleic acids (RNAs) have diverse applications, including imaging RNA trafficking and as a component of RNA-based sensors that exhibit fluorescence upon binding small molecules in live cells. These RNAs include the Spinach and Spinach2 aptamers, which bind and activate the fluorescence of fluorophores similar to that found in green fluorescent protein. Although additional highly fluorescent RNA-fluorophore complexes would extend the utility of this technology, the identification of novel RNA-fluorophore complexes is difficult. Current approaches select aptamers on the basis of their ability to bind fluorophores, even though fluorophore binding alone is not sufficient to activate fluorescence. Additionally, aptamers require extensive mutagenesis to efficiently fold and exhibit fluorescence in living cells. Here we describe a platform for rapid generation of highly fluorescent RNA-fluorophore complexes that are optimized for function in cells. This procedure involves selection of aptamers on the basis of their binding to fluorophores, coupled with fluorescence-activated cell sorting (FACS) of millions of aptamers expressed in Escherichia coli. Promising aptamers are then further optimized using a FACS-based directed evolution approach. Using this approach, we identified several novel aptamers, including a 49-nt aptamer, Broccoli. Broccoli binds and activates the fluorescence of (Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-1,2-dimethyl-1H-imidazol-5(4H)-one. Broccoli shows robust folding and green fluorescence in cells, and increased fluorescence relative to Spinach2. This reflects, in part, improved folding in the presence of low cytosolic magnesium concentrations. Thus, this novel fluorescence-based selection approach simplifies the generation of aptamers that are optimized for expression and performance in living cells. PMID:25337688

  7. Fluorescent detection of ATP based on signaling DNA aptamer attached silica nanoparticles

    NASA Astrophysics Data System (ADS)

    Wang, Yanyan; Wang, Yusong; Liu, Bin

    2008-10-01

    Novel methods for rapid, sensitive and low-cost biomolecule detection have attracted particular interest because of their wide use in medical diagnostics, food inspection and biomedical research applications. In this work, we report a simple and efficient silica nanoparticle (NP)-based fluorescent assay for ATP detection. It takes advantage of the washing and separation properties of NPs and the structure-switch property of DNA aptamers, resulting in fluorescence change of the supernatant in the presence of targets. A linear response for ATP detection was observed from 0 to 6 mM with a detection limit of ~34 µM. This detection strategy could be generalized to other aptamer-based detection systems.

  8. A peptide-based fluorescent chemosensor for multianalyte detection.

    PubMed

    Wang, Peng; Liu, Lixuan; Zhou, Panpan; Wu, Wenyu; Wu, Jiang; Liu, Weisheng; Tang, Yu

    2015-10-15

    A novel multifunctional peptide fluorescent chemosensor (DP-3) with a lysine backbone and double sides conjugated with histidine and dansyl groups has been designed and synthesized by solid phase synthesis. This chemosensor is a promising analytical tool for detecting Zn(2+), Cu(2+), and S(2-) based on different mechanisms in 100% aqueous solutions, and intracellular biosensing has been successfully actualized. The peptide beacon structure of DP-3 makes it more stable and capable of achieving multianalyte detection, especially for sulfide ions. Until now, there have been few examples of using a peptide fluorescent chemosensor to detect anions with a continuous method. As designed, DP-3 exhibits excellent cell permeation and low biotoxicity and displays high selectivity and sensitivity, with Zn(2+) and Cu(2+) detection limits of 82 nM and 78 nM, respectively. This study raises the new possibility of a highly selective peptide fluorescent chemosensor for multifunctional detection, including cation and anions, by different mechanisms in environmental and biological systems. We expect that this work will inspire the development of a multifunctional beacon peptide-based fluorescent chemosensor library using modifiable lateral and terminal groups for a variety of practical applications in physiological and pathological events. PMID:25957834

  9. Colorful Protein-Based Fluorescent Probes for Collagen Imaging

    PubMed Central

    Aper, Stijn J. A.; van Spreeuwel, Ariane C. C.; van Turnhout, Mark C.; van der Linden, Ardjan J.; Pieters, Pascal A.; van der Zon, Nick L. L.; de la Rambelje, Sander L.; Bouten, Carlijn V. C.; Merkx, Maarten

    2014-01-01

    Real-time visualization of collagen is important in studies on tissue formation and remodeling in the research fields of developmental biology and tissue engineering. Our group has previously reported on a fluorescent probe for the specific imaging of collagen in live tissue in situ, consisting of the native collagen binding protein CNA35 labeled with fluorescent dye Oregon Green 488 (CNA35-OG488). The CNA35-OG488 probe has become widely used for collagen imaging. To allow for the use of CNA35-based probes in a broader range of applications, we here present a toolbox of six genetically-encoded collagen probes which are fusions of CNA35 to fluorescent proteins that span the visible spectrum: mTurquoise2, EGFP, mAmetrine, LSSmOrange, tdTomato and mCherry. While CNA35-OG488 requires a chemical conjugation step for labeling with the fluorescent dye, these protein-based probes can be easily produced in high yields by expression in E. coli and purified in one step using Ni2+-affinity chromatography. The probes all bind specifically to collagen, both in vitro and in porcine pericardial tissue. Some first applications of the probes are shown in multicolor imaging of engineered tissue and two-photon imaging of collagen in human skin. The fully-genetic encoding of the new probes makes them easily accessible to all scientists interested in collagen formation and remodeling. PMID:25490719

  10. Fluorescent properties of merocyanines based on 1,3-indandione

    NASA Astrophysics Data System (ADS)

    Kulinich, A. V.; Mikitenko, E. K.; Ishchenko, A. A.

    2015-07-01

    We have studied the fluorescent properties of vinylogous series of merocyanine dyes based on 1,3-indandione and heterocycles of different electron-donating powers. Using a set of solvents with polarities lying in a wide range, we have analyzed the dependence of their solvatofluorochromism on the key structural parameters—the donor heterocyclic group and the length of the polymethine chain. It has been found that the signs of solvatochromism and solvatofluorochromism of merocyanines under study coincide. However, the solvent more weakly affects the position and the shape of their fluorescence bands than in the case of absorption spectra, especially, for negatively solvatochromic derivatives of 1,3-diphenylbenzimidazole. We have found that, upon passage from polar aprotic solvents to alcohols, the fluorescence quantum yields of dyes under investigation (irrespective of the sign of their solvatochromism) decrease. We have performed quantum-chemical calculations of merocyanine molecules by the DFT/B3LYP/6-31G(d,p) and TDDFT methods, including calculations taking into account the polarity of the medium by the PCM method. Based on the analysis of electronic transitions, we have explained the fluorescence quenching of indandione merocyanines in alcohol, which is unusual for carbonyl-containing intraionic dyes.

  11. The Fluorescence Enhancement of Mercury Detected in Food Based on Rhodamine Derivatives.

    PubMed

    Fan, Cai-ling; Xie, Pu-hui; Cui, Shu-min; Yang, Li-na; Sun, Qing; Ai, Zhi-lu

    2015-05-01

    Recently, the problem of food security is more and more serious, and people pay attention to mercury because of the toxic of it. A new approach for the determination of mercury content in foodstuff is devised. In this paper, first, we design and synthesis a new kind of fluorescent probe whose matrix based on rhodamine B, hydrazine hydrate and hydroxy benzaldehyde. Through the analysis of H-NMR spectra of the synthesized product L1, we confirm that the synthetic substance is the adjacent carboxyl benzaldehyde hydrazone structure generation of rhodamine B. Then, we measure the fluorescence signal intensity of the probe with different concentrations of mercury ions fully upon complexation by fluorescence spectrometer and we can study the relationship between the mercury ion concentration and the fluorescence intensity and draw the standard working curve. Following, It's time to discuss the microwave digestion processing of tea, after digestion we use the synthetic probe Li for determination of mercury content in tea. The experimental results show that the maximum excitation wavelength of the probe and coordination compound are 568. 05 and 560. 00 nm, the maximum emission wavelength are 587. 94 and 580. 00 nm. Then we can find the best testing conditions to improve the degree of accuracy, that is: room temperature, 50% the methanol solution, 3. 0 mL pH 4. 0 buffer solution, in the extent of 30 min. The experimental results show that Na+, K+, Ca2+, Cu2+, Zn2+, Al3+ have little impact on the fluorescence intensity of the:probe. Fe3+, Mg2+, Ba2+ has a weak enhancement to the fluorescence intensity of the probe. While a low concentrations of Hg2+ have an obviously enhanced effect on the fluorescence intensity of the probe. In contrast to other metal ions, the probe for Hg2+ has a good selectivity. Linear relationship between the magnitude of increase in fluorescence intensity and concentration of mercury ion was in the range of 5~20 ng . L-1 with detection limit (3S/N) of 1. 9 ng . L-1. The proposed method was applied to determination of mercury ion in samples of tea and sausage and the obtained result and sample recovery were all satisfactory. The methods of analysis instrument has the advantages of simple structure, sensitivity, high accuracy, good selectivity and less volume of simple, without the need for enrichment, being very practical. PMID:26415448

  12. Simple structured hybrid WOLEDs based on incomplete energy transfer mechanism: from blue exciplex to orange dopant

    NASA Astrophysics Data System (ADS)

    Zhang, Tianyou; Zhao, Bo; Chu, Bei; Li, Wenlian; Su, Zisheng; Yan, Xingwu; Liu, Chengyuan; Wu, Hairuo; Gao, Yuan; Jin, Fangming; Hou, Fuhua

    2015-05-01

    Exciplex is well known as a charge transfer state formed between electron-donating and electron-accepting molecules. However, exciplex based organic light emitting diodes (OLED) often performed low efficiencies relative to pure phosphorescent OLED and could hardly be used to construct white OLED (WOLED). In this work, a new mechanism is developed to realize efficient WOLED with extremely simple structure by redistributing the energy of triplet exciplex to both singlet exciplex and the orange dopant. The micro process of energy transfer could be directly examined by detailed photoluminescence decay measurement and time resolved photoluminescence analysis. This strategy overcomes the low reverse intersystem crossing efficiency of blue exciplex and complicated device structure of traditional WOLED, enables us to achieve efficient hybrid WOLEDs. Based on this mechanism, we have successfully constructed both exciplex-fluorescence and exciplex-phosphorescence hybrid WOLEDs with remarkable efficiencies.

  13. Simple structured hybrid WOLEDs based on incomplete energy transfer mechanism: from blue exciplex to orange dopant.

    PubMed

    Zhang, Tianyou; Zhao, Bo; Chu, Bei; Li, Wenlian; Su, Zisheng; Yan, Xingwu; Liu, Chengyuan; Wu, Hairuo; Gao, Yuan; Jin, Fangming; Hou, Fuhua

    2015-01-01

    Exciplex is well known as a charge transfer state formed between electron-donating and electron-accepting molecules. However, exciplex based organic light emitting diodes (OLED) often performed low efficiencies relative to pure phosphorescent OLED and could hardly be used to construct white OLED (WOLED). In this work, a new mechanism is developed to realize efficient WOLED with extremely simple structure by redistributing the energy of triplet exciplex to both singlet exciplex and the orange dopant. The micro process of energy transfer could be directly examined by detailed photoluminescence decay measurement and time resolved photoluminescence analysis. This strategy overcomes the low reverse intersystem crossing efficiency of blue exciplex and complicated device structure of traditional WOLED, enables us to achieve efficient hybrid WOLEDs. Based on this mechanism, we have successfully constructed both exciplex-fluorescence and exciplex-phosphorescence hybrid WOLEDs with remarkable efficiencies. PMID:25975371

  14. Simple structured hybrid WOLEDs based on incomplete energy transfer mechanism: from blue exciplex to orange dopant

    PubMed Central

    Zhang, Tianyou; Zhao, Bo; Chu, Bei; Li, Wenlian; Su, Zisheng; Yan, Xingwu; Liu, Chengyuan; Wu, Hairuo; Gao, Yuan; Jin, Fangming; Hou, Fuhua

    2015-01-01

    Exciplex is well known as a charge transfer state formed between electron-donating and electron-accepting molecules. However, exciplex based organic light emitting diodes (OLED) often performed low efficiencies relative to pure phosphorescent OLED and could hardly be used to construct white OLED (WOLED). In this work, a new mechanism is developed to realize efficient WOLED with extremely simple structure by redistributing the energy of triplet exciplex to both singlet exciplex and the orange dopant. The micro process of energy transfer could be directly examined by detailed photoluminescence decay measurement and time resolved photoluminescence analysis. This strategy overcomes the low reverse intersystem crossing efficiency of blue exciplex and complicated device structure of traditional WOLED, enables us to achieve efficient hybrid WOLEDs. Based on this mechanism, we have successfully constructed both exciplex-fluorescence and exciplex-phosphorescence hybrid WOLEDs with remarkable efficiencies. PMID:25975371

  15. Colorimetric and Fluorescent Sensing of SCN- Based on meso-Tetraphenylporphyrin/meso-Tetraphenylporphyrin Cobalt(II) System

    PubMed Central

    Zhang, Ying; Wang, Hua; Yang, Rong H.

    2007-01-01

    An approach for colorimetric and fluorescent sensing of thiocyanate (SCN-) has been proposed based on the competitive-displacement strategy between meso-tetraphenylporphyrin (TPP) and meso-tetraphenylporphyrin cobalt(II) (CoTPP). In THF-water solution, TPP emits strong fluorescence at 651 nm; however, the fluorescence was quenched stepwise by CoTPP, and then restored by SCN-, the detection limit is 6.0 10-4 M. The recognition of SCN- could also be easily achieved by visual way since the assembly system showed significant color change by the anion. Both the fluorescence and the color change of the system exhibits remarkably high selectivity to SCN- over a large series of anions. The interaction mechanisms among TPP, CoTPP and SCN- were primarily investigated by fluorescence lifetime. The quenching of TPP fluorescence is attributed to the formation of TPP/CoTPP aggregates, and the fluorescence restoration is due to the binding of CoTPP with SCN-, releasing the free TPP. This simple system has the potential to be used as a latent fluorescent sensing approach for SCN- for environmental analysis.

  16. Cell uptake and localization studies of squaramide based fluorescent probes.

    PubMed

    Sampedro, Angel; Villalonga-Planells, Ruth; Vega, Manuel; Ramis, Guillem; Fernndez de Mattos, Silvia; Villalonga, Priam; Costa, Antoni; Rotger, Carmen

    2014-08-20

    Cell internalization is a major issue in drug design. Although squaramide-based compounds are receiving much attention because of their interesting bioactivity, cell uptake and trafficking within cells of this type of compounds are still unknown. In order to monitor the cell internalization process of cyclosquaramide compounds we have prepared two fluorescent probes by covalently linking a fluorescent dye (BODIPY derivative or fluorescein) to a noncytotoxic cyclosquaramide framework. These two probes (C2-BDP and C2-FITC) rapidly internalize across live cell membranes through endocytic receptor-mediated mechanisms. Due to its higher fluorescence and photochemical stability, C2-BDP is a superior dye than C2-FITC. C2-BDP remains sequestered in late endosomes allowing their fast and selective imaging in various live cell types. Cyclosquaramide-cell membrane interactions facilitate cell uptake and have been investigated by binding studies in solution as well as in live cells. Cyclosquaramide 1 (C2-BDP) can be used as a highly fluorescent probe for the rapid and selective imaging of late endosomes in live cells. PMID:25036647

  17. Immunosensor systems with the Langmuir-film-based fluorescence detection

    SciTech Connect

    Chudinova, G K; Nagovitsyn, I A; Savranskii, V V; Karpov, R E

    2003-09-30

    A method is developed for detecting protein antigens for fluorescent immunoassay using a model system based on the technique for preparation of Langmuir films. Fluorescein isothiocyanate and donor-acceptor energy-transfer pairs of markers (the Yb complex of tetraphenyl porphyrin - benzoyl trifluoroacetoneisothiocyanate and derivatives of tetra(carboxyphenyl) porphyrin - cyanine dye containing a five-membered polyene chain), which were nor studied earlier, were used as markers for detecting the binding of an antigen on the surface of Langmuir films of antibodies. Fluorescence was detected in the near-IR region (for the first pair) and in the visible spectral range (for the second pair). To reduce the nonspecific sorption of a protein (antigen), a method was proposed for the preparation of a nonpolar surface by applying an even number of layers of stearic acid as a substrate for the Langmuir - Blodgett film. A high sensitivity of model systems to a protein antigen in solution was achieved ({approx}10{sup -11} M), the assay time being 6 - 8 min. The model system with the first donor - acceptor pair was tested in analysis of the blood plasma. The fluorescence of the Dy{sup 3+}, Tm{sup 3+}, and Yb{sup 3+} complexes of tetraphenyl porphyrin sensitised by diketonate complexes of lanthanides was studied for the first time and the enhancement of the IR fluorescence of these complexes in a Langmuir film was demonstrated. (papers devoted to the memory of academician a m prokhorov)

  18. Neurotransmitter imaging in living cells based on native fluorescence detection

    SciTech Connect

    Tan, W.; Yeung, E.S. |; Parpura, V.; Haydon, P.G.

    1995-08-01

    A UV laser-based optical microscope and CCD detection system with high sensitivity has been developed to image neurotransmitters in living cells. We demonstrate the detection of serotonin that has been taken up into individual living glial cells (astrocytes) based on its native fluorescence. We found that the fluorescence intensity of astrocytes increased by up to 10 times after serotonin uptake. The temporal resolution of this detection system at 10{sup -4} M serotonin is as fast as 50 ms, and the spatial resolution is diffraction limited. This UV laser microscope imaging system shows promise for studies of spatial-temporal dynamics of neurotransmitter levels in living neurons and glia. 19 refs., 5 figs., 1 tab.

  19. A simple, rapid, and high-throughput fluorescence polarization immunoassay for simultaneous detection of organophosphorus pesticides in vegetable and environmental water samples

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A simple, rapid, and high-throughput fluorescent polarization immunoassay (FPIA) for simultaneous determination of organophosphorus pesticides (OPs) was developed. Three haptens were labeled with a fluorescein probe and used as tracers to develop a homogenous FPIA using a broad-specificity monoclon...

  20. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography-part 2: image reconstruction.

    PubMed

    Correia, Teresa; Koch, Maximilian; Ale, Angelique; Ntziachristos, Vasilis; Arridge, Simon

    2016-02-21

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. We propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. Furthermore, structural information can be incorporated into the image reconstruction with PAD-WT to improve image quality and resolution. In this case, the weights used to average voxels in the image are calculated using the structural image, instead of the fluorescence image. The regularisation strength depends on both structural and fluorescence images, which guarantees that the method can preserve fluorescence information even when it is not structurally visible in the anatomical images. In part 1, we tested the method using a denoising problem. Here, we use simulated and in vivo mouse fDOT data to assess the algorithm performance. Our results show that the proposed PAD-WT method provides high quality and noise free images, superior to those obtained using AD. PMID:26808190

  1. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography—part 2: image reconstruction

    NASA Astrophysics Data System (ADS)

    Correia, Teresa; Koch, Maximilian; Ale, Angelique; Ntziachristos, Vasilis; Arridge, Simon

    2016-02-01

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. We propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. Furthermore, structural information can be incorporated into the image reconstruction with PAD-WT to improve image quality and resolution. In this case, the weights used to average voxels in the image are calculated using the structural image, instead of the fluorescence image. The regularisation strength depends on both structural and fluorescence images, which guarantees that the method can preserve fluorescence information even when it is not structurally visible in the anatomical images. In part 1, we tested the method using a denoising problem. Here, we use simulated and in vivo mouse fDOT data to assess the algorithm performance. Our results show that the proposed PAD-WT method provides high quality and noise free images, superior to those obtained using AD.

  2. Simple, Scalable, Script-Based Science Processor (S4P)

    NASA Technical Reports Server (NTRS)

    Lynnes, Christopher; Vollmer, Bruce; Berrick, Stephen; Mack, Robert; Pham, Long; Zhou, Bryan; Wharton, Stephen W. (Technical Monitor)

    2001-01-01

    The development and deployment of data processing systems to process Earth Observing System (EOS) data has proven to be costly and prone to technical and schedule risk. Integration of science algorithms into a robust operational system has been difficult. The core processing system, based on commercial tools, has demonstrated limitations at the rates needed to produce the several terabytes per day for EOS, primarily due to job management overhead. This has motivated an evolution in the EOS Data Information System toward a more distributed one incorporating Science Investigator-led Processing Systems (SIPS). As part of this evolution, the Goddard Earth Sciences Distributed Active Archive Center (GES DAAC) has developed a simplified processing system to accommodate the increased load expected with the advent of reprocessing and launch of a second satellite. This system, the Simple, Scalable, Script-based Science Processor (S42) may also serve as a resource for future SIPS. The current EOSDIS Core System was designed to be general, resulting in a large, complex mix of commercial and custom software. In contrast, many simpler systems, such as the EROS Data Center AVHRR IKM system, rely on a simple directory structure to drive processing, with directories representing different stages of production. The system passes input data to a directory, and the output data is placed in a "downstream" directory. The GES DAAC's Simple Scalable Script-based Science Processing System is based on the latter concept, but with modifications to allow varied science algorithms and improve portability. It uses a factory assembly-line paradigm: when work orders arrive at a station, an executable is run, and output work orders are sent to downstream stations. The stations are implemented as UNIX directories, while work orders are simple ASCII files. The core S4P infrastructure consists of a Perl program called stationmaster, which detects newly arrived work orders and forks a job to run the appropriate executable (registered in a configuration file for that station). Although S4P is written in Perl, the executables associated with a station can be any program that can be run from the command line, i.e., non-interactively. An S4P instance is typically monitored using a simple Graphical User Interface. However, the reliance of S4P on UNIX files and directories also allows visibility into the state of stations and jobs using standard operating system commands, permitting remote monitor/control over low-bandwidth connections. S4P is being used as the foundation for several small- to medium-size systems for data mining, on-demand subsetting, processing of direct broadcast Moderate Resolution Imaging Spectroradiometer (MODIS) data, and Quick-Response MODIS processing. It has also been used to implement a large-scale system to process MODIS Level 1 and Level 2 Standard Products, which will ultimately process close to 2 TB/day.

  3. A fluorescence enhancement-based sensor for hydrogen sulfate ion.

    PubMed

    Yang, Shih-Tse; Liao, De-Jhong; Chen, Shau-Jiun; Hu, Ching-Han; Wu, An-Tai

    2012-04-01

    Sugar-aza-crown ether-based cavitand 1 can act as a selective turn-on fluorescence sensor for hydrogen sulfate ion in methanol among a series of tested anions. Spectroscopic studies, particularly NMR spectroscopy, revealed that the C-H hydrogen bonding between 1,2,3-triazole ring of cavitand 1 and hydrogen sulfate ion is crucial for the high selectivity of the receptor for hydrogen sulfate. PMID:22363932

  4. A simple and effective method to synthesize fluorescent nanoparticles using tryptophan and light and their lethal effect against bacteria.

    PubMed

    Tomita, Rafael Jun; de Matos, Ricardo Almeida; Vallim, Marcelo Afonso; Courrol, Lilia Coronato

    2014-11-01

    A simple, environmentally friendly and cost-effective method was used to synthesize silver nanoparticles using tryptophan and light. To prepare the nanoparticles, the following components were used: deionized water, silver nitrate, light and tryptophan. The effects of the tryptophan concentration and light exposure time on the formation of tryptophan silver nanoparticles (Tnnps) were studied. The synthesized Tnnps were characterized using transmission electron microscopy (TEM), absorption and fluorescence spectroscopy and zeta potential measurements. The synthesized Tnnps were nearly spherical, with sizes of approximately 17 nm. In addition, the antibacterial activity of Tnnps was determined by monitoring the growth curves of strains of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, Serratia marcescens, and Enterococcus faecalis using the microdilution test. The Minimum Inhibitory Concentration (MIC) for 4 of 5 tested bacteria was determined to be between 20.0 and 17.5 ?g/mL for 48 h and between 22.5 and 20.0 ?g/mL for 72 h. PMID:25129701

  5. Simple, Rapid Mycobacterium ulcerans Disease Diagnosis from Clinical Samples by Fluorescence of Mycolactone on Thin Layer Chromatography

    PubMed Central

    Wadagni, Anita; Frimpong, Michael; Phanzu, Delphin Mavinga; Ablordey, Anthony; Kacou, Emmanuel; Gbedevi, Mirabelle; Marion, Estelle; Xing, Yalan; Babu, Vaddela Sudheer; Phillips, Richard Odame; Wansbrough-Jones, Mark; Kishi, Yoshito; Asiedu, Kingsley

    2015-01-01

    Introduction Mycobacterium ulcerans infection, known as Buruli ulcer, is a disease of the skin and subcutaneous tissues which is an important but neglected tropical disease with its major impact in rural parts of West and Central Africa where facilities for diagnosis and management are poorly developed. We evaluated fluorescent thin layer chromatography (f-TLC) for detection of mycolactone in the laboratory using samples from patients with Buruli ulcer and patients with similar lesions that gave a negative result on PCR for the IS2404 repeat sequence of M. ulcerans Methodology/Principal findings Mycolactone and DNA extracts from fine needle aspiration (FNA), swabs and biopsy specimen were used to determine the sensitivity and specificity of f-TLC when compared with PCR for the IS2404. For 71 IS2404 PCR positive and 28 PCR negative samples the sensitivity was 73.2% and specificity of 85.7% for f-TLC. The sensitivity was similar for swabs (73%), FNAs (75%) and biopsies (70%). Conclusions We have shown that mycolactone can be detected from M. ulcerans infected skin tissue by f-TLC technique. The technique is simple, easy to perform and read with minimal costs. In this study it was undertaken by a member of the group from each endemic country. It is a potentially implementable tool at the district level after evaluation in larger field studies. PMID:26583925

  6. Paper-based upconversion fluorescence resonance energy transfer biosensor for sensitive detection of multiple cancer biomarkers.

    PubMed

    Xu, Sai; Dong, Biao; Zhou, Donglei; Yin, Ze; Cui, Shaobo; Xu, Wen; Chen, Baojiu; Song, Hongwei

    2016-01-01

    A paper-based upconversion fluorescence resonance energy transfer assay device is proposed for sensitive detection of CEA. The device is fabricated on a normal filter paper with simple nano-printing method. Upconversion nanoparticles tagged with specific antibodies are printed to the test zones on the test paper, followed by the introduction of assay antigen. Upconversion fluorescence measurements are directly conducted on the test zones after the antigen-to-antibody reactions. Furthermore, a multi-channel test paper for simultaneous detection of multiple cancer biomarkers was established by the same method and obtained positive results. The device showed high anti-interfere, stability, reproducible and low detection limit (0.89 ng/mL), moreover it is very easy to fabricate and operate, which is a promising prospect for a clinical point-of-care test. PMID:27001460

  7. A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences

    NASA Astrophysics Data System (ADS)

    Klamp, Tobias; Yahiatne, Idir; Lampe, Andr; Schttpelz, Mark; Sauer, Markus

    2011-03-01

    The sensitive and rapid detection of pathogenic DNA is of tremendous importance in the field of diagnostics. We demonstrate the ability of detecting and quantifying single- and double-stranded pathogenic DNA with picomolar sensitivity in a bead-based fluorescence assay. Selecting appropriate capturing and detection sequences enables rapid (2 h) and reliable DNA quantification. We show that synthetic sequences of S. pneumoniae and M. luteus can be quantified in very small sample volumes (20 ?L) across a linear detection range over four orders of magnitude from 1 nM to 1 pM, using a miniaturized wide-field fluorescence microscope without amplification steps. The method offers single molecule detection sensitivity without using complex setups and thus volunteers as simple, robust, and reliable method for the sensitive detection of DNA and RNA sequences.

  8. A coumarin-indole based colorimetric and 'turn on' fluorescent probe for cyanide

    NASA Astrophysics Data System (ADS)

    Xu, Yu; Dai, Xi; Zhao, Bao-Xiang

    2015-03-01

    A novel coumarin-indole based chemodosimeter with a simple structure was designed and prepared via a condensation reaction in high yield. The probe exhibited very high selectivity towards cyanide on both fluorescence and UV-vis spectra, which allowed it to quantitatively detect and imaging cyanide ions in organic-aqueous solution by either fluorescence enhancement or colorimetric changes. Confirmed by 1H NMR and HRMS spectra, the detection mechanism was proved to be related with the Michael addition reaction induced by cyanide ions, which blocked the intramolecular charge transfer (ICT) of the probe. Moreover, the probe was able to be utilized efficiently in a wide pH range (7.5-10) with negligible interference from other anions and a low detection limit of 0.51 μM. Application in 5 kinds of natural water source and accurate detection of cyanide in tap water solvent system also indicated the high practical significance of the probe.

  9. Paper-based upconversion fluorescence resonance energy transfer biosensor for sensitive detection of multiple cancer biomarkers

    PubMed Central

    Xu, Sai; Dong, Biao; Zhou, Donglei; Yin, Ze; Cui, Shaobo; Xu, Wen; Chen, Baojiu; Song, Hongwei

    2016-01-01

    A paper-based upconversion fluorescence resonance energy transfer assay device is proposed for sensitive detection of CEA. The device is fabricated on a normal filter paper with simple nano-printing method. Upconversion nanoparticles tagged with specific antibodies are printed to the test zones on the test paper, followed by the introduction of assay antigen. Upconversion fluorescence measurements are directly conducted on the test zones after the antigen-to-antibody reactions. Furthermore, a multi-channel test paper for simultaneous detection of multiple cancer biomarkers was established by the same method and obtained positive results. The device showed high anti-interfere, stability, reproducible and low detection limit (0.89 ng/mL), moreover it is very easy to fabricate and operate, which is a promising prospect for a clinical point-of-care test. PMID:27001460

  10. Construction of an off-on fluorescence system based on carbon dots for trace pyrophosphate sensing.

    PubMed

    Yue, Qiaoli; Hou, Yining; Yue, Shuzhen; Du, Kaimin; Shen, Tongfei; Wang, Lei; Xu, Shuling; Li, Haibo; Liu, Jifeng

    2015-05-01

    A novel and simple fluorescence Off-On system is proposed for selective pyrophosphate (PPi) sensing in an aqueous solution. The method is constructed based on the strong blue emission of carbon dots (CDs) owing to its outstanding photoluminescence and easy synthesis, which has shown exciting potential in analytical and biological field. The fluorescence of CDs can be remarkably quenched by some transition metal ions such as Cu(2+), Ni(2+), Mn(2+) and Co(2+) due to the coordination reaction between metal ions and the carboxylic groups on the surface of CDs. When PPi was introduced to CDs-metal ion system the fluorescence of CDs was recovered regularly. The increment of fluorescence intensity was proportional with the concentration of PPi in the range of 1-200?M and correspondingly the limit of detection was calculated as 0.32?M according to the recommendation of IUPAC as 3.29 S B/m. The possible mechanism was discussed for the detection of PPi and the quenching reaction between CDs and metal ions. Furthermore, the proposed system was successfully used to monitor the content of PPi in water samples from artificial wetland. PMID:25731813

  11. Determination of adenine based on the fluorescence recovery of the L-Tryptophan-Cu2+ complex

    NASA Astrophysics Data System (ADS)

    Duan, Ruilin; Li, Chunyan; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Yuan, Yusheng; Hu, Xiaoli

    2016-01-01

    A simple and sensitive method for determination of adenine was developed based on fluorescence quenching and recovery of L-Tryptophan (L-Trp). The fluorescence of L-Trp could efficiently quenched by copper ion compared with other common metal ions. Upon addition of adenine (Ade) in L-Trp-Cu(II) system, the fluorescence was reoccurred. Under the optimum conditions, the recovery fluorescence intensity was linearly correlated with the concentration of adenine in the range from 0.34 to 25.0 μmol L-1, with a correlation coefficient (R2) of 0.9994. The detection limit (3σ/k) was 0.046 μmol L-1, indicating that this method could applied to detect trace adenine. In this study, amino acids including L-Trp, D-Trp, L-Tyr, D-Tyr, L-Phe, D-Phe were investigated and only L-Trp could well chelated copper ion. Additionally, the mechanism of quench and recovery also were discussed and the method was successfully applied to detect the adenine in DNA with satisfactory results.

  12. A novel fluorescence assay for inorganic pyrophosphatase based on modulated aggregation of graphene quantum dots.

    PubMed

    Zhu, Xueli; Liu, Jinwen; Peng, Haiyang; Jiang, Jianhui; Yu, Ruqin

    2016-01-01

    A simple and highly sensitive fluorometric method has been developed for inorganic pyrophosphatase (PPase) activity detection based on the disaggregation and aggregation of graphene quantum dots (GQDs). Copper ions can trigger the severe aggregation of GQDs with rich carboxyl groups, which results in effective fluorescence quenching. While, with the addition of pyrophosphate (PPi), the quenched fluorescence is effectively recovered owing to the strong interaction between PPi and Cu(2+). Furthermore, under the catalytic hydrolysis of PPase, the complex of PPi-Cu(2+)-PPi is rapidly disassembled, and the fluorescence is re-quenched. This method is highly sensitive and selective for PPase detection, with a linear correlation between the fluorescence intensity and the PPase concentration in the range from 1 to 200 mU mL(-1) with a detection limit down to 1 mU mL(-1) (S/N = 3). Additionally, the inhibition effect of NaF on the PPase activity is also studied. Thus, the proposed method may hold a potential application in the diagnosis of PPase-related diseases and screening of PPase inhibitors, to evaluate the function and inhibition of PPase in biological systems. PMID:26581179

  13. Determination of adenine based on the fluorescence recovery of the L-Tryptophan-Cu(2+) complex.

    PubMed

    Duan, Ruilin; Li, Chunyan; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Yuan, Yusheng; Hu, Xiaoli

    2016-01-01

    A simple and sensitive method for determination of adenine was developed based on fluorescence quenching and recovery of L-Tryptophan (L-Trp). The fluorescence of L-Trp could efficiently quenched by copper ion compared with other common metal ions. Upon addition of adenine (Ade) in L-Trp-Cu(II) system, the fluorescence was reoccurred. Under the optimum conditions, the recovery fluorescence intensity was linearly correlated with the concentration of adenine in the range from 0.34 to 25.0?molL(-1), with a correlation coefficient (R(2)) of 0.9994. The detection limit (3?/k) was 0.046?molL(-1), indicating that this method could applied to detect trace adenine. In this study, amino acids including L-Trp, D-Trp, L-Tyr, D-Tyr, L-Phe, D-Phe were investigated and only L-Trp could well chelated copper ion. Additionally, the mechanism of quench and recovery also were discussed and the method was successfully applied to detect the adenine in DNA with satisfactory results. PMID:26225734

  14. High efficient solar tracker based on a simple shutter structure

    NASA Astrophysics Data System (ADS)

    Chen, Jin-Jia; Liu, Te-Shu; Huang, Kuang-Lung; Lin, Po-Chih

    2013-09-01

    In many photovoltaic (PV) or sunlight-illumination systems, solar trackers are always essential to obtain high energy/flux concentration efficiency, and that would lead to increase cost and extra power consumption due to the complex structure and heavy weight of the trackers. To decrease the cost while without sacrificing efficiency, a Fresnellens concentrator incorporated with a simple and cheap shutter, which consists of high reflective mirrors instead of conventional trackers, is proposed in this paper to provide solar tracking during the daytime. Thus, the time-variant and slant-incident sunlight rays can be redirected to vertically incident upon the surface of the Fresnel lens by appropriately arranging mirrors and swinging them to the proper slant angles with respect to the orientation of sunlight. The computer simulation results show that power concentration efficiency over 90%, as compared with the efficiency of directly normal incident sunlight, can be achieved with the mirror reflectance of 0.97 and for any solar incident angle within +/-75 degrees to the normal of the Fresnel lens. To verify the feasibility and performance of the concentrator with the proposed shutter, a sunlight illumination system based on this novel structure is demonstrated. Both computer simulation and practical measurement results for the prototype of the sunlight illumination system are also given to compare with. The results prove the simple and high efficient shutter applicable to general PV or sunlight-illumination systems for solar tracking.

  15. Wide and scalable field-of-view Talbot-grid-based fluorescence microscopy

    PubMed Central

    Pang, Shuo; Han, Chao; Kato, Mihoko; Sternberg, Paul W.; Yang, Changhuei

    2013-01-01

    Here we report a low-cost and simple wide field-of-view (FOV) on-chip fluorescence-imaging platform, termed fluorescence Talbot microscopy (FTM), which utilizes the Talbot self-imaging effect to enable efficient fluorescence imaging over a large and directly scalable FOV. The FTM prototype has a resolution of 1.2 ?m and an FOV of 3.9 mm 3.5 mm. We demonstrate the imaging capability of FTM on fluorescently labeled breast cancer cells (SK-BR-3) and human embryonic kidney 293 (HEK) cells expressing green fluorescent protein. PMID:23202123

  16. Nanoparticle-based energy transfer for rapid and simple detection of protein glycosylation

    SciTech Connect

    Oh, Eunkeu; Lee, Dohoon; Kim, Young-Pil; Cha, Seung YOUP; Oh, Doo BEYONG; Kim, Jungbae; Kang, Hyun AH; Kim, Hak SUNG

    2006-12-04

    Glycan moiety of glycoproteins plays an essential role in its biological activity in vivo, and the analysis of glycosylation is of great importance in the development of protein therapeutics. In this study, we report a rapid and simple detection of protein glycosylation based on the fluorescence resonance energy transfer (FRET) between concanavalin A-conjugated gold nanoparticles (ConA-AuNPs) and dextran-conjugated quantum dots (Dex-QDs). The increased photoluminescence (PL) signals of Dex-QDs due to the competitive inhibition of glycoproteins were well correlated with the glycosylation chain length of glucose oxidases as well as the mannosylation degree of bovine serum albumin (BSA). The parallel analysis of the diversely mannosylated BSAs using an image analyzer further demonstrated the potential of this new technique in high-throughput screening of glycoprotein and carbohydrate therapeutics.

  17. Simple-random-sampling-based multiclass text classification algorithm.

    PubMed

    Liu, Wuying; Wang, Lin; Yi, Mianzhu

    2014-01-01

    Multiclass text classification (MTC) is a challenging issue and the corresponding MTC algorithms can be used in many applications. The space-time overhead of the algorithms must be concerned about the era of big data. Through the investigation of the token frequency distribution in a Chinese web document collection, this paper reexamines the power law and proposes a simple-random-sampling-based MTC (SRSMTC) algorithm. Supported by a token level memory to store labeled documents, the SRSMTC algorithm uses a text retrieval approach to solve text classification problems. The experimental results on the TanCorp data set show that SRSMTC algorithm can achieve the state-of-the-art performance at greatly reduced space-time requirements. PMID:24778587

  18. Monitoring methionine sulfoxide with stereospecific mechanism-based fluorescent sensors

    PubMed Central

    Tarrago, Lionel; Péterfi, Zalán; Lee, Byung Cheon; Michel, Thomas; Gladyshev, Vadim N.

    2015-01-01

    Methionine can be reversibly oxidized to methionine sulfoxide (MetO) under physiological and pathophysiological conditions, but its use as a redox marker suffers from the lack of tools to detect and quantify MetO within cells. In this work, we created a pair of complementary stereospecific genetically-encoded mechanism-based ratiometric fluorescent sensors of MetO by inserting a circularly yellow fluorescent protein between yeast methionine sulfoxide reductases and thioredoxins. The two sensors, named MetSOx and MetROx for their ability to detect S and R-forms of MetO, respectively, were utilized for targeted analysis of protein oxidation, regulation and repair, as well as for monitoring MetO in bacterial and mammalian cells, analyzing compartment-specific changes in MetO, and examining responses to physiological stimuli. PMID:25799144

  19. Monitoring methionine sulfoxide with stereospecific mechanism-based fluorescent sensors.

    PubMed

    Tarrago, Lionel; Pterfi, Zaln; Lee, Byung Cheon; Michel, Thomas; Gladyshev, Vadim N

    2015-05-01

    Methionine can be reversibly oxidized to methionine sulfoxide (MetO) under physiological and pathophysiological conditions, but its use as a redox marker suffers from the lack of tools to detect and quantify MetO within cells. In this work, we created a pair of complementary stereospecific genetically encoded mechanism-based ratiometric fluorescent sensors of MetO by inserting a circularly permuted yellow fluorescent protein between yeast methionine sulfoxide reductases and thioredoxins. The two sensors, respectively named MetSOx and MetROx for their ability to detect S and R forms of MetO, were used for targeted analysis of protein oxidation, regulation and repair as well as for monitoring MetO in bacterial and mammalian cells, analyzing compartment-specific changes in MetO and examining responses to physiological stimuli. PMID:25799144

  20. Hybridization assay based on evanescent fluorescence excitation and collection

    NASA Astrophysics Data System (ADS)

    Sumner, James J.; Mmerole, Robert U.; Stratis-Cullum, Dimitra N.; Yi, Hyunmin; Bentley, William E.; Gillespie, James B.

    2003-08-01

    There is a great need for high throughput and sensitive sensors for genetic analysis. These sensors can be used for varied purposes from monitoring gene expression in organims to speciation of possible pathogens. Consequently, an instrument capable of these tasks would be a great benefit for food and water safety, medical diagnostics and defense of military and civilian populations from biological threats. This work examines the development of a hybridization-based biosensor using a novel tapered fiber optic rpobe. The immobilization of single-stranded, synthetic ologinucleotides utilizing aminoproplytriethoxysilane and glutaraldehyde was implemented on the fiber optic sensor. Hybridization takes place with a complementary analyte sequence followed by a fluorescent, labeled signaling probe to form a sandwich assay. Following hybridization, the fiber is interrogated with a diode laser source and the resulting fluorescence signal is detected using a miniature spectrometer.

  1. A study of boronic acid based fluorescent glucose sensors.

    PubMed

    Kawanishi, T; Romey, M A; Zhu, P C; Holody, M Z; Shinkai, S

    2004-09-01

    Boronic acid based anthracene dyes were designed, synthesized, and immobilized to solid phase, creating a continuous glucose sensor. Glucose sensitivities of dyes can decrease drastically after immobilization, therefore how to immobilize a dye to solid phase without changing the dye property is a key issue in developing the sensor. The glucose sensitivity of the simplest 1st generation sensor, which is based on an immobilized mono-phenylboronate/single-arm type, came short of the sensitivity requirement for practical use, because of the very moderate fluorescence intensity change over the physiological glucose range. However, the 2nd generation, an immobilized bis-phenylboronate/double-arm type sensor, which contained two boronate groups in the dye moiety in expectation of a large intensity change, brought about considerable improvement on its glucose sensitivity. We tried to introduce functional groups onto an anthracene ring in order to improve the dies' fluorescence properties. Acetyl or carboxyl substitution on anthracene contributed to shift the fluorescence wavelength into the more visible range (red-shift) and a divergence of wavelength between an excitation peak and an emission peak. This improvement is advantageous to the design of an optical detection system. Furthermore, single arm immobilization to this carboxyl group, thus linking directly to the fluorophore led to a 3rd generation sensor, an immobilized bis-phenylboronate/single-arm type, that was twice as sensitive as that of the 2nd generation sensor, presumably due to increased mobility of the dye moiety. The results of our study advance closer toward a clinically useful continuous fluorescent glucose sensor. PMID:15617258

  2. Simple and highly sensitive determination of free fatty acids in human serum by high performance liquid chromatography with fluorescence detection.

    PubMed

    Iwata, T; Inoue, K; Nakamura, M; Yamaguchi, M

    1992-01-01

    A highly sensitive and simple reversed phase high performance liquid chromatographic (HPLC) method for the quantitative determination of free fatty acids in human serum is presented. The method is based on the direct derivatization of serum fatty acids with 6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone-3-propionylcarboxylic acid hydrazide. The derivatization reaction proceeds in aqueous solution in the presence of pyridine and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at 37 degrees C. The resulting derivatives are separated within 75 min on a reversed phase column (YMC Pack C8) with a gradient elution of aqueous acetonitrile and detected fluorimetrically. The detection limits are 2.5-5 fmol in a 10 microL injection volume. The sensitivity permits precise determination of free fatty acids in 5 microL serum. The method is simple and is without the conventional liquid-liquid extraction steps of serum fatty acids. PMID:1525484

  3. Chemical Cytometry: Fluorescence-Based Single-Cell Analysis

    NASA Astrophysics Data System (ADS)

    Cohen, Daniella; Dickerson, Jane A.; Whitmore, Colin D.; Turner, Emily H.; Palcic, Monica M.; Hindsgaul, Ole; Dovichi, Norman J.

    2008-07-01

    Cytometry deals with the analysis of the composition of single cells. Flow and image cytometry employ antibody-based stains to characterize a handful of components in single cells. Chemical cytometry, in contrast, employs a suite of powerful analytical tools to characterize a large number of components. Tools have been developed to characterize nucleic acids, proteins, and metabolites in single cells. Whereas nucleic acid analysis employs powerful polymerase chain reaction-based amplification techniques, protein and metabolite analysis tends to employ capillary electrophoresis separation and ultrasensitive laser-induced fluorescence detection. It is now possible to detect yoctomole amounts of many analytes in single cells.

  4. A simple microviscometric approach based on Brownian motion tracking

    NASA Astrophysics Data System (ADS)

    Hnyluchov, Zuzana; Bjalon?kov, Petra; Karas, Pavel; Mravec, Filip; Halasov, Tereza; Peka?, Miloslav; Kubala, Luk; Vte?ek, Jan

    2015-02-01

    Viscosityan integral property of a liquidis traditionally determined by mechanical instruments. The most pronounced disadvantage of such an approach is the requirement of a large sample volume, which poses a serious obstacle, particularly in biology and biophysics when working with limited samples. Scaling down the required volume by means of microviscometry based on tracking the Brownian motion of particles can provide a reasonable alternative. In this paper, we report a simple microviscometric approach which can be conducted with common laboratory equipment. The core of this approach consists in a freely available standalone script to process particle trajectory data based on a Newtonian model. In our study, this setup allowed the sample to be scaled down to 10 ?l. The utility of the approach was demonstrated using model solutions of glycerine, hyaluronate, and mouse blood plasma. Therefore, this microviscometric approach based on a newly developed freely available script can be suggested for determination of the viscosity of small biological samples (e.g., body fluids).

  5. A simple microviscometric approach based on Brownian motion tracking.

    PubMed

    Hnyluchov, Zuzana; Bjalon?kov, Petra; Karas, Pavel; Mravec, Filip; Halasov, Tereza; Peka?, Miloslav; Kubala, Luk; Vte?ek, Jan

    2015-02-01

    Viscosity-an integral property of a liquid-is traditionally determined by mechanical instruments. The most pronounced disadvantage of such an approach is the requirement of a large sample volume, which poses a serious obstacle, particularly in biology and biophysics when working with limited samples. Scaling down the required volume by means of microviscometry based on tracking the Brownian motion of particles can provide a reasonable alternative. In this paper, we report a simple microviscometric approach which can be conducted with common laboratory equipment. The core of this approach consists in a freely available standalone script to process particle trajectory data based on a Newtonian model. In our study, this setup allowed the sample to be scaled down to 10 ?l. The utility of the approach was demonstrated using model solutions of glycerine, hyaluronate, and mouse blood plasma. Therefore, this microviscometric approach based on a newly developed freely available script can be suggested for determination of the viscosity of small biological samples (e.g., body fluids). PMID:25725855

  6. A simple plasmid-based transient gene expression method using High Five cells.

    PubMed

    Shen, Xiao; Pitol, Ana K; Bachmann, Virginie; Hacker, David L; Baldi, Lucia; Wurm, Florian M

    2015-12-20

    The High Five (H5) cell line, derived from the lepidopteran Trichoplusia ni, is one of the major insect cell hosts for the production of recombinant proteins using the baculovirus expression vector system (BEVS). Here, we describe a simple polyethylenimine (PEI)-based transient gene expression (TGE) process for the rapid production of recombinant proteins from suspension-adapted H5 cells. The method was optimized using two model proteins, enhanced green fluorescent protein (EGFP) and human tumor necrosis factor receptor-Fc fusion protein (TNFR-Fc). After screening several promoter and enhancer combinations for high levels of TNFR:Fc production, an expression vector containing the Autographa californica multicapsid nucleopolyhedrovirus immediate early 1 (ie1) promoter and homologous region 5 (hr5) enhancer was selected. Cells were transfected at a density of 210(6) cells/mL by direct addition of DNA and PEI. Under optimized conditions, a 90% transfection efficiency (percentage of EGFP-positive cells) was obtained. In addition, we observed volumetric TNFR-Fc yields over 150?g/mL within 4 days of transfection. The method was found to be reproducible and scalable to 300mL. This plasmid-based transient transfection process is a simple and efficient alternative to the BEVS for recombinant protein production in H5 cells. PMID:26476358

  7. Fluorescence-based detection of single-nucleotide changes in RNA using graphene oxide and DNAzyme.

    PubMed

    Hong, Chaesun; Kim, Dong-Min; Baek, Ahruem; Chung, Hyewon; Jung, Woong; Kim, Dong-Eun

    2015-04-01

    We report a simple fluorometric method for detection of single-nucleotide changes in RNA using graphene oxide (GO) and RNA-cleaving DNAzyme. The fluorescent DNA probe (F-DNA) was annealed to RNA fragments generated by RNA cleavage with DNAzyme specific to mutant RNA. The F-DNA-RNA duplex attenuated the quenching of F-DNA fluorescence by GO. PMID:25714982

  8. CINCH (confocal incoherent correlation holography) super resolution fluorescence microscopy based upon FINCH (Fresnel incoherent correlation holography)

    PubMed Central

    Siegel, Nisan; Storrie, Brian; Bruce, Marc

    2016-01-01

    FINCH holographic fluorescence microscopy creates high resolution super-resolved images with enhanced depth of focus. The simple addition of a real-time Nipkow disk confocal image scanner in a conjugate plane of this incoherent holographic system is shown to reduce the depth of focus, and the combination of both techniques provides a simple way to enhance the axial resolution of FINCH in a combined method called “CINCH”. An important feature of the combined system allows for the simultaneous real-time image capture of widefield and holographic images or confocal and confocal holographic images for ready comparison of each method on the exact same field of view. Additional GPU based complex deconvolution processing of the images further enhances resolution. PMID:26839443

  9. Photon upconversion in homogeneous fluorescence-based bioanalytical assays.

    PubMed

    Soukka, Tero; Rantanen, Terhi; Kuningas, Katri

    2008-01-01

    Upconverting phosphors (UCPs) are very attractive reporters for fluorescence resonance energy transfer (FRET)-based bioanalytical assays. The large anti-Stokes shift and capability to convert near-infrared to visible light via sequential absorption of multiple photons enable complete elimination of autofluorescence, which commonly impairs the performance of fluorescence-based assays. UCPs are ideal donors for FRET, because their very narrow-banded emission allows measurement of the sensitized acceptor emission, in principle, without any crosstalk from the donor emission at a wavelength just tens of nanometers from the emission peak of the donor. In addition, acceptor dyes emitting at visible wavelengths are essentially not excited by near-infrared, which further emphasizes the unique potential of upconversion FRET (UC-FRET). These characteristics result in favorable assay performance using detection instrumentation based on epifluorometer configuration and laser diode excitation. Although UC-FRET is a recently emerged technology, it has already been applied in both immunoassays and nucleic acid hybridization assays. The technology is also compatible with optically difficult biological samples, such as whole blood. Significant advances in assay performance are expected using upconverting lanthanide-doped nanocrystals, which are currently under extensive research. UC-FRET, similarly to other fluorescence techniques based on resonance energy transfer, is strongly distance dependent and may have limited applicability, for example in sandwich-type assays for large biomolecules, such as viruses. In this article, we summarize the essentials of UC-FRET, describe its current applications, and outline the expectations for its future potential. PMID:18596348

  10. A Simple, Scalable, Script-based Science Processor

    NASA Technical Reports Server (NTRS)

    Lynnes, Christopher

    2004-01-01

    The production of Earth Science data from orbiting spacecraft is an activity that takes place 24 hours a day, 7 days a week. At the Goddard Earth Sciences Distributed Active Archive Center (GES DAAC), this results in as many as 16,000 program executions each day, far too many to be run by human operators. In fact, when the Moderate Resolution Imaging Spectroradiometer (MODIS) was launched aboard the Terra spacecraft in 1999, the automated commercial system for running science processing was able to manage no more than 4,000 executions per day. Consequently, the GES DAAC developed a lightweight system based on the popular Per1 scripting language, named the Simple, Scalable, Script-based Science Processor (S4P). S4P automates science processing, allowing operators to focus on the rare problems occurring from anomalies in data or algorithms. S4P has been reused in several systems ranging from routine processing of MODIS data to data mining and is publicly available from NASA.

  11. Cyanine-based probe\\tag-peptide pair fluorescence protein imaging and fluorescence protein imaging methods

    DOEpatents

    Mayer-Cumblidge, M. Uljana; Cao, Haishi

    2013-01-15

    A molecular probe comprises two arsenic atoms and at least one cyanine based moiety. A method of producing a molecular probe includes providing a molecule having a first formula, treating the molecule with HgOAc, and subsequently transmetallizing with AsCl.sub.3. The As is liganded to ethanedithiol to produce a probe having a second formula. A method of labeling a peptide includes providing a peptide comprising a tag sequence and contacting the peptide with a biarsenical molecular probe. A complex is formed comprising the tag sequence and the molecular probe. A method of studying a peptide includes providing a mixture containing a peptide comprising a peptide tag sequence, adding a biarsenical probe to the mixture, and monitoring the fluorescence of the mixture.

  12. A colorimetric and fluorescent cyanide chemosensor based on dicyanovinyl derivatives: Utilization of the mechanism of intramolecular charge transfer blocking

    NASA Astrophysics Data System (ADS)

    Li, Qiao; Cai, Yi; Yao, Hong; Lin, Qi; Zhu, Yuan-Rong; Li, Hui; Zhang, You-Ming; Wei, Tai-Bao

    2015-02-01

    Chemosensor (CS1) was designed and synthesized by simple green chemistry procedure. CS1 exhibited both colorimetric and fluorescence turn-off responses for cyanide (CN-) ion in aqueous solution. The probe showed an immediate visible color changes from yellow to colorless and green fluorescence disappearance when CN- was added. The mechanism of chemosensor reaction with CN- was studied using 1HH NMR and 13C NMR spectroscopies and mass spectrometry. Moreover, test strips based on the sensor were fabricated, which served as convenient and efficient CN- test kits.

  13. Fluorescent nanosensor for probing histone acetyltransferase activity based on acetylation protection and magnetic graphitic nanocapsules.

    PubMed

    Han, Yitao; Li, Pei; Xu, Yiting; Li, Hao; Song, Zhiling; Nie, Zhou; Chen, Zhuo; Yao, Shouzhuo

    2015-02-18

    Protein acetylation catalyzed by histone acetyltransferases (HATs) is significant in biochemistry and pharmacology because of its crucial role in epigenetic gene regulations. Herein, an antibody-free fluorescent nanosensor is developed for the facile detection of HAT activity based on acetylation protection against exopeptidase cleavage and super-quenching ability of nanomaterials. It is shown for the first time that HAT-catalyzed acetylation could protect the peptide against exopeptidase digestion. FITC-tagged acetylated peptide causes the formation of a nano-quenchers/peptide nano-complex resulting in fluorescence quenching, while the unacetylated peptide is fully degraded by exopeptidase to release the fluorophore and restore fluorescence. Four kinds of nano-quenchers, including core-shell magnetic graphitic nanocapsules (MGN), graphene oxide (GO), single-walled carbon nanotubes (SWCNTs), and gold nanoparticles (AuNPs), are comprehensively compared. MGN shows the best selectivity to recognize the acetylated peptide and the lowest detection limit because of its excellent quenching efficiency and magnetic enrichment property. With this MGN-based nanosensor, HAT p300 is detected down to 0.1 nM with wide linear range from 0.5 to 100 nM. This sensor is feasible to assess HAT inhibition and detect p300 activity in cell lysate. The proposed nanosensor is simple, sensitive, and cost-effective for HAT assay, presenting a promising toolkit for epigenetic research and HAT-targeted drug discovery. PMID:25277402

  14. Effects of Mechanical Constraint on the Performance of Fluorescent Hydrogel-based Fiber Optic Sensors

    NASA Astrophysics Data System (ADS)

    Jukl, Jennifer Marie

    Although biosensor technology is a broad and well-studied field, the progress of many novel sensor technologies faces challenges. These challenges range from simple design considerations to fundamental issues with the concept or approach. One of the most active fields of sensor research integrates fiber optics with specially engineered fluorescent molecules. This type of sensor typically utilizes a porous polymer or porous glass substrate to entrap the fluorescent (or fluorescently-tagged) molecule. Porous polymer hydrogels are generally favored due to their ease of fabrication, low cost, adaptability, and biocompatibility. While hydrogels are ideal for both functional molecule suspension and fluid diffusion, their porosity and hydrophilicity are not always advantageous. The largest drawback of these properties is the hydrogel swelling they produce and the resulting geometric changes. This project investigated the limitations of fluorescent hydrogel-based sensors and the effects of unpredictable structural changes hydrogels undergo during typical, unrestrained swelling. The significance of covalent incorporation of the sensing fluorophore into the hydrogel matrix is also explored. Leaching tests were conducted using polyacrylamide (PAm) hydrogels which were impregnated with one of two pH sensitive fluorophores, one which bonded covalently with the hydrogel matrix during polymerization (fluorescein o-acrylate), and one which did not (fluorescein sodium). Once determined to be effective, the covalently bonding fluorophore was used to create constrained-dimension fluorescent pH sensors. These sensors were tested for effectiveness and reproducibility. All data was collected using a laboratory grade optical fibers, a USB spectrometer, and SpectraSuite software (Ocean Optics, 2010) unless otherwise specified.

  15. Ditopic pillar[5]arene-based fluorescence enhancement material mediated by [c2]daisy chain formation.

    PubMed

    Wang, Kai; Wang, Chun-Yu; Zhang, Yumo; Zhang, Sean Xiao-An; Yang, Bing; Yang, Ying-Wei

    2014-08-28

    Ditopic pillarene-based fluorescence enhancement material induced by [c2]daisy chain formation has been successfully fabricated. The highly fluorescent smart material exhibits great adaptivity and can act as a multi-responsive supramolecular sensor. PMID:25008450

  16. Cell-Based Lipid Flippase Assay Employing Fluorescent Lipid Derivatives.

    PubMed

    Jensen, Maria S; Costa, Sara; Gnther-Pomorski, Thomas; Lpez-Marqus, Rosa L

    2016-01-01

    P-type ATPases in the P4 subfamily (P4-ATPases) are transmembrane proteins unique for eukaryotes that act as lipid flippases, i.e., to translocate phospholipids from the exofacial to the cytofacial monolayer of cellular membranes. While initially characterized as aminophospholipid translocases, studies of individual P4-ATPase family members from fungi, plants, and animals show that P4-ATPases differ in their substrate specificities and mediate transport of a broader range of lipid substrates. Here, we describe an assay based on fluorescent lipid derivatives to monitor and characterize lipid flippase activities in the plasma membrane of cells, using yeast as an example. PMID:26695048

  17. Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone

    SciTech Connect

    Moester, Martiene J.C.; Schoeman, Monique A.E.; Oudshoorn, Ineke B.; Percuros BV, Leiden ; Beusekom, Mara M. van; Mol, Isabel M.; Percuros BV, Leiden ; Kaijzel, Eric L.; Löwik, Clemens W.G.M.; Rooij, Karien E. de; Percuros BV, Leiden

    2014-01-03

    Highlights: •We validate a simple and fast method of quantification of in vitro mineralization. •Fluorescently labeled agents can detect calcium deposits in the mineralized matrix of cell cultures. •Fluorescent signals of the probes correlated with Alizarin Red S staining. -- Abstract: Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments. KS483 cells were cultured under osteogenic conditions in the presence of compounds that either stimulate or inhibit osteoblast differentiation and thereby matrix mineralization. After 21 days of differentiation, fluorescence of stained cultures was quantified with a near-infrared imager and compared to Alizarin Red S quantification. Fluorescence of both probes closely correlated to Alizarin Red S staining in both inhibiting and stimulating conditions. In addition, both compounds displayed specificity for mineralized nodules. We therefore conclude that this method of quantification of bone mineralization using fluorescent compounds is a good alternative for the Alizarin Red S staining.

  18. Expression-Enhanced Fluorescent Proteins Based on Enhanced Green Fluorescent Protein for Super-resolution Microscopy.

    PubMed

    Duwé, Sam; De Zitter, Elke; Gielen, Vincent; Moeyaert, Benjamien; Vandenberg, Wim; Grotjohann, Tim; Clays, Koen; Jakobs, Stefan; Van Meervelt, Luc; Dedecker, Peter

    2015-10-27

    "Smart fluorophores", such as reversibly switchable fluorescent proteins, are crucial for advanced fluorescence imaging. However, only a limited number of such labels is available, and many display reduced biological performance compared to more classical variants. We present the development of robustly photoswitchable variants of enhanced green fluorescent protein (EGFP), named rsGreens, that display up to 30-fold higher fluorescence in E. coli colonies grown at 37 °C and more than 4-fold higher fluorescence when expressed in HEK293T cells compared to their ancestor protein rsEGFP. This enhancement is not due to an intrinsic increase in the fluorescence brightness of the probes, but rather due to enhanced expression levels that allow many more probe molecules to be functional at any given time. We developed rsGreens displaying a range of photoswitching kinetics and show how these can be used for multimodal diffraction-unlimited fluorescence imaging such as pcSOFI and RESOLFT, achieving a spatial resolution of ∼70 nm. By determining the first ever crystal structures of a negative reversibly switchable FP derived from Aequorea victoria in both the "on"- and "off"-conformation we were able to confirm the presence of a cis-trans isomerization and provide further insights into the mechanisms underlying the photochromism. Our work demonstrates that genetically encoded "smart fluorophores" can be readily optimized for biological performance and provides a practical strategy for developing maturation- and stability-enhanced photochromic fluorescent proteins. PMID:26308583

  19. Sensitive turn-on fluorescent detection of tartrazine based on fluorescence resonance energy transfer.

    PubMed

    Huang, Sheng Tian; Shi, Yan; Li, Nian Bing; Luo, Hong Qun

    2012-01-18

    We introduce a sensitive, rapid, label-free and general fluorescent method for the determination of tartrazine by competitive binding to reduced graphene oxide (rGO) against fluorescein, and the fluorescence recovery upon fluorescein desorption from rGO provides a quantitative readout for tartrazine, giving a detection limit of 0.53 ng mL(-1). PMID:22121502

  20. Rapid fluorescence-based measurement of toxicity in anaerobic digestion.

    PubMed

    Chen, Jian Lin; Ortiz, Raphael; Xiao, Yeyuan; Steele, Terry W J; Stuckey, David C

    2015-05-15

    A rapid fluorescence measurement based on resazurin reduction was developed and applied for the detection of toxicants/inhibitors to anaerobic digestion metabolism. By initially using a pure facultative anaerobic strain, Enterococcus faecalis as a model organism, this technique proved to be fast and sensitive when detecting the model toxicant, pentachlorophenol (PCP). The technique revealed significant metabolic changes in Enterococcus faecalis with a PCP spike ranging from 0.05 to 100 mg/L, and could detect PCP's toxicity to E. faecalis at a concentration of only 0.05 mg/L in 8 min. Furthermore, by extending this technique to a mixed anaerobic sludge, not only could the effect of 0.05-100 mg/L PCP be determined on anaerobic digestion metabolism within 10 min, but also its rate of biogas production. These results suggest that a resazurin-based fluorescence measurement can potentially be incorporated into a microfluidic system to develop a biosensor for the real-time monitoring, control and early warning of toxicant/inhibitor loads in the influent to an anaerobic digestion system. PMID:25768985

  1. A VBA-based Simulation for Teaching Simple Linear Regression

    ERIC Educational Resources Information Center

    Jones, Gregory Todd; Hagtvedt, Reidar; Jones, Kari

    2004-01-01

    In spite of the name, simple linear regression presents a number of conceptual difficulties, particularly for introductory students. This article describes a simulation tool that provides a hands-on method for illuminating the relationship between parameters and sample statistics.

  2. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography-part 1: technical principles.

    PubMed

    Correia, Teresa; Arridge, Simon

    2016-02-21

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. In this two-part paper, we propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. The proposed method combines the nonlocal means (NLM), AD and wavelet shrinkage methods, which are image processing methods. Therefore, in this first paper, we used a denoising test problem to analyse the performance of the new method. Our results show that the proposed PAD-WT method provides better results than the AD or NLM methods alone. The efficacy of the method for fDOT image reconstruction problem is evaluated in part 2. PMID:26808057

  3. Patch-based anisotropic diffusion scheme for fluorescence diffuse optical tomography—part 1: technical principles

    NASA Astrophysics Data System (ADS)

    Correia, Teresa; Arridge, Simon

    2016-02-01

    Fluorescence diffuse optical tomography (fDOT) provides 3D images of fluorescence distributions in biological tissue, which represent molecular and cellular processes. The image reconstruction problem is highly ill-posed and requires regularisation techniques to stabilise and find meaningful solutions. Quadratic regularisation tends to either oversmooth or generate very noisy reconstructions, depending on the regularisation strength. Edge preserving methods, such as anisotropic diffusion regularisation (AD), can preserve important features in the fluorescence image and smooth out noise. However, AD has limited ability to distinguish an edge from noise. In this two-part paper, we propose a patch-based anisotropic diffusion regularisation (PAD), where regularisation strength is determined by a weighted average according to the similarity between patches around voxels within a search window, instead of a simple local neighbourhood strategy. However, this method has higher computational complexity and, hence, we wavelet compress the patches (PAD-WT) to speed it up, while simultaneously taking advantage of the denoising properties of wavelet thresholding. The proposed method combines the nonlocal means (NLM), AD and wavelet shrinkage methods, which are image processing methods. Therefore, in this first paper, we used a denoising test problem to analyse the performance of the new method. Our results show that the proposed PAD-WT method provides better results than the AD or NLM methods alone. The efficacy of the method for fDOT image reconstruction problem is evaluated in part 2.

  4. Introducing a fluorescence-based standard to quantify protein partitioning into membranes.

    PubMed

    Thomas, Franziska A; Visco, Ilaria; Petrek, Zden?k; Heinemann, Fabian; Schwille, Petra

    2015-11-01

    The affinity of peripheral membrane proteins for a lipid bilayer can be described using the partition coefficient (KP). Although several methods to determine KP are known, all possess limitations. To address some of these issues, we developed both: a versatile method based on single molecule detection and fluorescence imaging for determining KP, and a simple measurement standard employing hexahistidine-tagged enhanced green fluorescent protein (eGFP-His6) and free standing membranes of giant unilamellar vesicles (GUVs) functionalized with NTA(Ni) lipids as binding sites. To ensure intrinsic control, our method features two measurement modes. In the single molecule mode, fluorescence correlation spectroscopy (FCS) is applied to quantify free and membrane associated protein concentrations at equilibrium and calculate KP. In the imaging mode, confocal fluorescence images of GUVs are recorded and analyzed with semi-automated software to extract protein mean concentrations used to derive KP. Both modes were compared by determining the affinity of our standard, resulting in equivalent KP values. As observed in other systems, eGFP-His6 affinity for membranes containing increasing amounts of NTA(Ni) lipids rises in a stronger-than-linear fashion. We compared our dual approach with a FCS-based assay that uses large unilamellar vesicles (LUVs), which however fails to capture the stronger-than-linear trend for our NTA(Ni)-His6 standard. Hence, we determined the KP of the MARCKS effector domain with our FCS approach on GUVs, whose results are consistent with previously published data using LUVs. We finally provide a practical manual on how to measure KP and understand it in terms of molecules per lipid surface. PMID:26342678

  5. A sensitive detection of T4 polynucleotide kinase activity based on ?-cyclodextrin polymer enhanced fluorescence combined with an exonuclease reaction.

    PubMed

    Song, Chunxia; Yang, Xiaohai; Wang, Kemin; Wang, Qing; Liu, Jianbo; Huang, Jin; He, Leiliang; Liu, Pei; Qing, Zhihe; Liu, Wei

    2015-02-01

    A strategy for T4 polynucleotide kinase activity detection was proposed based on a ?-cyclodextrin polymer (poly?-CD) and an exonuclease reaction. The fluorescence of pyrene enhanced by more than 10 times in the presence of poly?-CD, and a simple detection of T4 PNK was achieved with a detection limit of 0.02 units per mL. PMID:25519768

  6. A simple, low-cost, remote fiber-optic micro volume fluorescence flowcell for capillary flow-injection analysis.

    PubMed

    Hart, Sean J; Jiji, Renee D

    2002-10-01

    A small volume flowcell for fluorescence detection in capillary flow injection (CFI) analysis has been created by using a low cost, commercially available fluidic device. Fluorescence detection is achieved using an optical fiber to deliver excitation light to the sample flowing through the device and another optical fiber to collect fluorescence emission. The flowcell is a standard fluidic cross with a swept volume of 721 nL. Optical fibers were oriented at right angles using standard sleeves and ferrules to set their position near the cross intersection. Multiple excitation sources were used including a low power UV laser and blue and UV light emitting diodes (LED). The full emission spectrum detection limits, using the laser, for fluorescein and bovine serum albumin (BSA) were 0.30 ppb and 2.1 x 10(-4)% (w/w), respectively. Two fluidic crosses were used in series for multi-wavelength fluorescence excitation using fiber-optically coupled LED. PMID:12373382

  7. Monitoring water supplies for weaponized bacteria and bacterial toxins using rapid fluorescence-based viability and affinity assays

    NASA Astrophysics Data System (ADS)

    Van Tassell, Roger L.; Evans, Mishell

    2004-03-01

    The rapid detection of weaponized bacteria and toxins is a major problem during a biological attack. Although sensitive detection formats exist for many biowarfare agents, they often require advanced training and complex procedures. Luna has developed simple, rapid means for determining the presence of pathogens and bacterial toxins in water supplies using fluorescence-based assays that can be adapted for field use. The batteries of rapid assays are designed for i) determining cell viability and bacterial loads by exploiting metabolic markers (e.g., acid-production, redox potentials, etc) and ii) detecting bacterial toxins using fluorescent, polymerized affinity liposomes (fluorosomes). The viability assays were characterized using E. coli, S. aureus and the anthrax simulant, B. globigii. The viability assays detected bacterial loads of ~ 104 CFU/ml and with simple filtration ~ 100CFU/ml could be detected. The affinity fluorosomes were characterized using cholera toxin (CT). Affinity liposomes displaying GM1 and anti-CT antibodies could detect CT at fluorescence system, Luna characterized the binding of affinity fluorosomes to respective targets and determined the responses of bacterial loads in the fluorescent viability assays. Using this two-tiered approach, Luna demonstrated that water susceptible to sabotage could be easily monitored and confirmed for specific agents using simple, general and specific fluorescence-based detection schemes based on metabolism and ligand-target interactions.

  8. Physically-based in silico light sheet microscopy for visualizing fluorescent brain models

    PubMed Central

    2015-01-01

    Background We present a physically-based computational model of the light sheet fluorescence microscope (LSFM). Based on Monte Carlo ray tracing and geometric optics, our method simulates the operational aspects and image formation process of the LSFM. This simulated, in silico LSFM creates synthetic images of digital fluorescent specimens that can resemble those generated by a real LSFM, as opposed to established visualization methods producing visually-plausible images. We also propose an accurate fluorescence rendering model which takes into account the intrinsic characteristics of fluorescent dyes to simulate the light interaction with fluorescent biological specimen. Results We demonstrate first results of our visualization pipeline to a simplified brain tissue model reconstructed from the somatosensory cortex of a young rat. The modeling aspects of the LSFM units are qualitatively analysed, and the results of the fluorescence model were quantitatively validated against the fluorescence brightness equation and characteristic emission spectra of different fluorescent dyes. AMS subject classification Modelling and simulation PMID:26329404

  9. Glycol Chitosan-Based Fluorescent Theranostic Nanoagents for Cancer Therapy

    PubMed Central

    Rhee, Jin-Kyu; Park, Ok Kyu; Lee, Aeju; Yang, Dae Hyeok; Park, Kyeongsoon

    2014-01-01

    Theranostics is an integrated nanosystem that combines therapeutics with diagnostics in attempt to develop new personalized treatments with enhanced therapeutic efficacy and safety. As a promising therapeutic paradigm with cutting-edge technologies, theranostic agents are able to simultaneously deliver therapeutic drugs and diagnostic imaging agents and also monitor the response to therapy. Polymeric nanosystems have been intensively explored for biomedical applications to diagnose and treat various cancers. In recent years, glycol chitosan-based nanoagents have been developed as dual-purpose materials for simultaneous diagnosis and therapy. They have shown great potential in cancer therapies, such as chemotherapeutics and nucleic acid and photodynamic therapies. In this review, we summarize the recent progress and potential applications of glycol chitosan-based fluorescent theranostic nanoagents for cancer treatments and discuss their possible underlying mechanisms. PMID:25522316

  10. Capillary electrophoresis based on continuous-wave multiphoton excitation fluorescence

    NASA Astrophysics Data System (ADS)

    Chen, Sheng; Luo, Qingming; Zeng, Shaoqun; Chen, Tongsheng; Gong, Hui; Fu, Ling

    2003-12-01

    High performance capillary electrophoresis(HPCE) has been widely applied to the field of chemistry and life science for its advantages of high separation efficiency, short analysis times and low sample volume requirements. But how to improve the detection capability of capillary electrophoresis was ever a key problem. Multi-photon excitation (MPE) fluorescence detection could solve this problem, however this method needed expensive femtosecond mod-locked laser, which restricted the improvement of capillary electrophoresis. In this paper, we implemented a set of MPE-CE system based on a continuous wave (CW), successfully exploited the CW multi-photon excitation to achieve detection of capillary electrophoresis. In our current MPE-CE system we used Ti:sapphire laser to prove MPE-CE could work in CW mode. From this point, we would substitute the laser with laser diode (LD) in future. This is really a new but also practical idea in MPE-CE system based on CW.

  11. Coordination polymer based on cyano: Synthesis, crystal structure, and fluorescence

    SciTech Connect

    Fang Zhenlan; He Jiangang; Ju Qiang; Wu Xiaoyuan; Lu Canzhong

    2010-07-15

    One novel 2-D polythreading framework named as [Cu{sub 3}(CN){sub 3}(NH{sub 3})] (1), was obtained through the self-assembling of CuCN under hydrothermal reaction. It is remarkable that there is a 26-membered [Cu{sub 10}(CN){sub 8}] decanuclear metallamacrocycle with the effective size of ca. 16.8x6.83 A{sup 2} along the a-axis. These 2-D layers stack in an ..ABAB...staggered fashion, with the lateral {l_brace}(CN)Cu{sub 3}(NH{sub 3}){r_brace} moieties of each layer inserting into the voids of the decanuclear metallamacrocycles from two adjacent layers. Optical diffuse reflectance spectrum and the result of DFT calculation reveal that 1 is potential direct semiconducting material. In the solid state at room temperature, 1 shows bright yellow fluorescence under ultraviolet light illumination. Its emissive excited state is primarily attributed to the LMCT, LLCT and {sup 3}[MMLCT] excited state, based on the result of DFPT calculation. - Graphical abstract: One novel 2-D polythreading framework [Cu{sub 3}(CN){sub 3}(NH{sub 3})] (1) obtained through the self-assembling of CuCN under hydrothermal reaction shows bright yellow fluorescence at room temperature.

  12. Wafer-scale aluminum plasmonics for fluorescence based biodetection

    NASA Astrophysics Data System (ADS)

    Farhang, Arash; George, Matthew C.; Williamson, Brent; Black, Mike; Wangensteen, Ted; Fraser, James; Petrova, Rumyana; Prestgard, Kent

    2015-08-01

    Moxtek has leveraged existing capabilities in wafer-scale patterning of sub-wavelength wire grid polarizers into the fabrication of 1D and 2D periodic aluminum plasmonic structures. This work will discuss progress in 200 mm diameter wafer-scale fabrication, with detailed emphasis within the realm of microarray based fluorescence detection. Aluminum nanohole arrays in a hexagonal lattice are first numerically investigated. The nanohole array geometry and periodicity are specifically tuned to coincide both with the excitation of the fluorophore Cy3, and to provide a high field enhancement within the nanoholes where labeled biomolecules are captured. This is accomplished through numerical modelling, nanofabrication, SEM imaging, and optical characterization. A 200mm diameter wafer, patterned with the optically optimized nanohole array, is cut into standard 1x3 inch microscope slide pieces and then subsequently printed with various antigens at 9 different concentrations. A sandwich bioassay is then carried out, using the corresponding conjugate antibodies in order to demonstrate specificity. The nanohole array exhibit a 3-4 times total fluorescence enhancement of Cy3, when compared to a leading commercial microarray glass slide.

  13. Optical biopsy fiber-based fluorescence spectroscopy instrumentation

    NASA Astrophysics Data System (ADS)

    Katz, Alvin; Ganesan, Singaravelu; Yang, Yuanlong; Tang, Gui C.; Budansky, Yury; Celmer, Edward J.; Savage, Howard E.; Schantz, Stimson P.; Alfano, Robert R.

    1996-04-01

    Native fluorescence spectroscopy of biomolecules has emerged as a new modality to the medical community in characterizing the various physiological conditions of tissues. In the past several years, many groups have been working to introduce the spectroscopic methods to diagnose cancer. Researchers have successfully used native fluorescence to distinguish cancerous from normal tissue samples in rat and human tissue. We have developed three generations of instruments, called the CD-scan, CD-ratiometer and CD-map, to allow the medical community to use optics for diagnosing tissue. Using ultraviolet excitation and emission spectral measurements on both normal and cancerous tissue of the breast, gynecology, colon, and aerodigestive tract can be separated. For example, from emission intensities at 340 nm to 440 nm (300 nm excitation), a statistically consistent difference between malignant tissue and normal or benign tissue is observed. In order to utilize optical biopsy techniques in a clinical setting, the CD-scan instrument was developed, which allows for rapid and reliable in-vitro and in-vivo florescence measurements of the aerodigestive tract with high accuracy. The instrumentation employs high sensitivity detection techniques which allows for lamp excitation, small diameter optical fiber probes; the higher spatial resolution afforded by the small diameter probes can increase the ability to detect smaller tumors. The fiber optic probes allow for usage in the aerodigestive tract, cervix and colon. Needle based fiber probes have been developed for in-vivo detection of breast cancer.

  14. A new turn-off fluorescence probe based on graphene quantum dots for detection of Au(III) ion

    NASA Astrophysics Data System (ADS)

    Amjadi, Mohammad; Shokri, Roghayeh; Hallaj, Tooba

    2016-01-01

    In this work, a new turn-off fluorescence probe based on the graphene quantum dots (GQDs) was designed for detection and quantification of Au(III) ion. GQDs were prepared by two simple carbonization methods using glucose (g-GQDs) and citric acid (c-GQDs) as carbon sources. The effect of some metal ions on the fluorescence intensity of the prepared GQDs was studied. It was found that the fluorescence of both GQDs is significantly quenched by Au(III) ions but the sensitivity and analytical performances are different for two prepared GQDs. Using g-GQDs, a new analytical method was developed for the determination of Au(III) in the concentration range of 1.0-80 ?M, with a detection limit of 0.5 ?M. The developed method was applied to the determination of Au(III) in water and plasma samples with satisfactory results.

  15. A new turn-off fluorescence probe based on graphene quantum dots for detection of Au(III) ion.

    PubMed

    Amjadi, Mohammad; Shokri, Roghayeh; Hallaj, Tooba

    2016-01-15

    In this work, a new turn-off fluorescence probe based on the graphene quantum dots (GQDs) was designed for detection and quantification of Au(III) ion. GQDs were prepared by two simple carbonization methods using glucose (g-GQDs) and citric acid (c-GQDs) as carbon sources. The effect of some metal ions on the fluorescence intensity of the prepared GQDs was studied. It was found that the fluorescence of both GQDs is significantly quenched by Au(III) ions but the sensitivity and analytical performances are different for two prepared GQDs. Using g-GQDs, a new analytical method was developed for the determination of Au(III) in the concentration range of 1.0-80?M, with a detection limit of 0.5?M. The developed method was applied to the determination of Au(III) in water and plasma samples with satisfactory results. PMID:26452097

  16. Microplate bioassay for nisin in foods, based on nisin-induced green fluorescent protein fluorescence.

    PubMed

    Reunanen, J; Saris, P E J

    2003-07-01

    A plasmid coding for the nisin two-component regulatory proteins, NisK and NisR, was constructed; in this plasmid a gfp gene (encoding the green fluorescent protein) was placed under control of the nisin-inducible nisF promoter. The plasmid was transformed into non-nisin-producing Lactococcus lactis strain MG1614. The new strain could sense extracellular nisin and transduce it to green fluorescent protein fluorescence. The amount of fluorescence was dependent on the nisin concentration, and it could be measured easily. By using this strain, an assay for quantification of nisin was developed. With this method it was possible to measure as little as 2.5 ng of pure nisin per ml in culture supernatant, 45 ng of nisin per ml in milk, 0.9 microg of nisin in cheese, and 1 microg of nisin per ml in salad dressings. PMID:12839802

  17. Frequency-domain flow cytometry: fluorescence-lifetime-based sensing technology for analyzing cells and chromosomes labeled with fluorescent probes

    NASA Astrophysics Data System (ADS)

    Steinkamp, John A.; Crissman, Harry A.; Lehnert, Bruce E.; Lehnert, Nancy M.; Deka, Chiranjit

    1997-05-01

    A flow cytometer has been developed that combines flow cytometry (FCM) and fluorescence lifetime spectroscopy measurement principles to provide unique capabilities for making frequency-domain, excited-state lifetime measurements on cells/chromosomes labeled with fluorescent probes, while preserving conventional FCM capabilities. Cells are analyzed as they intersect a high-frequency, intensity-modulated (sine-wave) laser excitation beam. Fluorescence signals are processed by (1) low-pass filtering to obtain conventional FCM dc-excited signals and (2) phase-sensitive detection electronics to resolve heterogeneous fluorescence based on differences in lifetimes expressed as phase-shifts and to quantify fluorescence lifetimes in real time. Processed signals are displayed as frequency distribution histograms and bivariate contour diagrams. Recent examples of biological applications include: (1) lifetime histograms recorded on autofluorescent human lung fibroblasts, murine thymus cells labeled with antibodies conjugated to fluorophores for studying fluorescence quenching as a function of antibody dilution and F/P ratio, and on cultured cells, nuclei, and chromosomes stained with DNA-binding fluorochromes and (2) phase-resolved, fluorescence signal- intensity histograms recorded on autofluorescent HLFs labeled with immunofluorescence markers and on murine thymus cells labeled with Red 613-antiThy 1.2 and propidium iodide (PI positive `dead' cells) to demonstrate the resolution of signals from highly overlapping emission spectra. This technology will increase the number of fluorescent markers usable in multilabeling studies and lifetimes can be used as spectroscopic probes to study the interaction of markers with their targets, each other, and the surrounding microenvironment.

  18. Autojoin: A Simple Rule Based Query Service for Complex Databases

    NASA Astrophysics Data System (ADS)

    Gaffney, N. I.; Gardner, L.; Brandt, M.

    Most databases used today are no longer flat. While the power of using these more complex data stores is well known, construction of queries can be quite a complex task. Currently this often requires detailed knowledge of the database structure and schema. As we move towards a VO paradigm, users cannot be expected to know the structure of databases, but will need to query them. Databases will need to provide query engines to complete queries automatically given only what the user wants to have returned and any qualifications they place on the query. For years StarView, a database query and data retrieval tool for the Space Telescope Science Institute, relied on a complex third party LISP-based program (QUICK) to construct valid SQL queries for the one database it could query. This limited our ability to support StarView as we could not easily add new rules to the system without completely rebuilding the query engine. Furthermore, QUICK did not have the ability to create SQL that would join tables in different databases (but hosted on the same server). Finally, the cost of upgrading to a new version of QUICK was prohibitively high. Our solution was to develop a rather simple database table driven Perl CGI program which is able to take as its input a skeleton SQL program. This may come from a program or other web page. In the query only the SELECT and user qualified WHERE clause are specified; no FROM or WHERE clause join information is included. The service then returns a fully qualified and syntactically correct query for the host database SQL program that can be used to get the information the user needs. Thus, an additional layer of abstraction for dealing with databases is created, freeing the user from having to know how tables are related in the database. In this paper we discuss the design and algorithm used to make Autojoin work as well as discuss how, when combined with a robust and searchable description of all the fields that can be publicly queried in the database, it allows users to tailor their questions to the database with ease and a high rate of success.

  19. Development of an image processing support system based on fluorescent dye to prevent elderly people with dementia from wandering.

    PubMed

    Nishigaki, Yutaka; Tanaka, Kentaro; Kim, Juhyon; Nakajima, Kazuki

    2013-01-01

    The wandering of elderly people with dementia is a significant behavioral problem and is a heavy burden on caregivers in residential and nursing homes. Thus, warning systems have been developed to prevent elderly people with dementia from leaving the premises. Some of these systems use radio waves. However, systems based on radio waves present several practical problems. For instance, the transmitter must be carried and may become lost; in addition, the battery of the transmitter must be changed. To solve these problems, we developed a support system that prevents elderly people with dementia from wandering. The system employs image processing technology based on fluorescent dye. The composition of the support system can be described as follows: fluorescent dye is painted in a simple shape on the clothes of an elderly person. The fluorescent color becomes visible by irradiation with a long wavelength of ultraviolet light. In the present paper, the relationship between the color of the dye and the cloth was investigated. A 3D video camera was used to acquire a 3D image and detect the simple shape. As a preliminary experiment, 3 colors (red, green and blue) of fluorescent dye were applied to cloths of 9 different colors. All fluorescent colors were detected on 6 of the cloths, but red and blue dye could not be detected on the other 3 cloths. In contrast, green dye was detectable on all 9 of the cloths. Additionally, we determined whether green dye could be detected in an actual environment. A rectangular shaped patch of green fluorescent dye was painted on the shoulder area of a subject, from the scapula to the clavicle. As a result, the green dye was detected on all 9 different colored cloths. PMID:24111431

  20. Fluorescence-based measurement of water-dissolved nitrate ions

    NASA Astrophysics Data System (ADS)

    Sharma, Ashutosh; Street, Nicolas J.

    1999-02-01

    A novel method for measuring ion concentration is reported based on a photochemical reaction between reduced nitrate ions and 2-amino-1-napthalene sulphonic acid. Reduced nitrate ions, in the form of nitrite, react with phot excited 2-amino-1- naphthalene sulphonic acid in acidic media resulting in the formation of a fluorescent product. The photochemical reaction was found selective to nitrite ions, with interference only from sulphide, sulphite, thiosulphate and iron (III) among various water dissolved ionic species investigated. The reported reaction offers a method to measure both nitrate and nitrite ion concentrations. The calibration plot was linear over the investigated range of 0 1 - 12 (mu) M and a detection limit of 24 nM plus or minus 2.4 nM. The method which was used to investigate nitrite ion concentrations in North London tap water and commercially available bottled water was found to be suitable for nitrate and nitrite ion measurement in such aqueous media.

  1. A simple but highly selective and sensitive fluorescence reporter for toxic CdII ion via excimer formation

    NASA Astrophysics Data System (ADS)

    Samanta, Anuva; Guchhait, Nikhil; Bhattacharya, Subhash Chandra

    2014-09-01

    High selectivity and fluorescence sensitivity of a well-recognized antiexcitotoxic and anticonvulsant drug, kynurenic acid (KA), toward CdII ion has been demonstrated by UV-vis, fluorescence, 1H NMR spectroscopy in combination with computational calculations. Upon complexation with CdII, KA exhibits a distinct excimer emission at 528 nm along with monomer emission at 402 nm. The sensing ability of drug KA toward CdII ion is distinctly different from sensing for HgII or CuII ion. KA can form dimer by intermolecular O-H⋯O hydrogen bonding between the -COOH groups. Presence of CdII metal ions promotes dimer formation in the excited state which exhibits excimer fluorescence.

  2. CuInS2 quantum dots-based fluorescence turn off/on probe for detection of melamine.

    PubMed

    Liu, Siyu; Hu, Junjie; Zhang, Hao; Su, Xingguang

    2012-11-15

    In this paper, a sensitive and simple method for the determination of melamine (MA) was developed based on the fluorescence changes of the water-soluble CuInS(2) quantum dots (QDs). The water-soluble CuInS(2) QDs capped by mercaptopropionic acid (MPA) was directly synthesized by hydrothermal method based on our previous report. The fluorescence emission of CuInS(2) QDs was quenched by the oxidation of the surface of the QDs with H(2)O(2), and the quenched fluorescence of CuInS(2) QDs could be recovered upon the addition of small amounts of MA, which might be due to the surface passivation of the CuInS(2) QDs by MA. The other amino acids such as glycine and lysine had no effect on the quenched fluorescence of CuInS(2) QDs. Under optimum conditions, there was a good linear relationship between the fluorescence intensity of CuInS(2) QDs and the concentration range of MA from 1.0×10(-8) to 1.0×10(-5) mol/L with a detection limit as low as 5 nM. The proposed method was successfully applied to detect trace MA in raw milk with satisfactory results. Compared with previous reports, the proposed method manifested several advantages such as high sensitivity, short analysis time, low cost and ease of operation. PMID:23158336

  3. A naphthalimide-based fluorescent sensor for halogenated solvents.

    PubMed

    Dai, Li; Wu, Di; Qiao, Qinglong; Yin, Wenting; Yin, Jun; Xu, Zhaochao

    2016-01-26

    A fluorescent sensor for halogenated solvents termed is reported. shows strong fluorescence in most halogenated solvents (QE > 0.1) but weak fluorescence (QE<0.01) in most non-halogenated solvents. In chlorinated solvents, the fluorescence intensity decreased with the reduction of chlorine content. On the contrary, in brominated solvents the fluorescence intensity increased with the reduction of bromine content. It is worth mentioning that displayed fluorescence emission centered at 520 nm in CCl4 with a quantum yield of 0.607, at 556 nm in CHCl3 with a quantum yield of 0.318, at 584 nm in CH2Cl2 with a quantum yield of 0.128, whereas in CHBr3 was centered at 441 nm with a quantum yield of 0.012. was shown to have the ability to differentiate CCl4, CHCl3, CH2Cl2 and CHBr3 halogenated solvents. PMID:26691881

  4. Latest methods of fluorescence-based protein crystal identification

    PubMed Central

    Meyer, Arne; Betzel, Christian; Pusey, Marc

    2015-01-01

    Successful protein crystallization screening experiments are dependent upon the experimenter being able to identify positive outcomes. The introduction of fluorescence techniques has brought a powerful and versatile tool to the aid of the crystal grower. Trace fluorescent labeling, in which a fluorescent probe is covalently bound to a subpopulation (<0.5%) of the protein, enables the use of visible fluorescence. Alternatively, one can avoid covalent modification and use UV fluorescence, exploiting the intrinsic fluorescent amino acids present in most proteins. By the use of these techniques, crystals that had previously been obscured in the crystallization drop can readily be identified and distinguished from amorphous precipitate or salt crystals. Additionally, lead conditions that may not have been obvious as such under white-light illumination can be identified. In all cases review of the screening plate is considerably accelerated, as the eye can quickly note objects of increased intensity. PMID:25664782

  5. Simple, benign, aqueous-based amination of polycarbonate surfaces

    DOE PAGESBeta

    VanDelinder, Virginia; Wheeler, David R.; Small, Leo J.; Brumbach, Michael T.; Spoerke, Erik D.; Henderson, Ian; Bachand, George D.

    2015-03-18

    Here we report a simple, safe, environmentally-friendly aqueous method that uses diamines to functionalize a polycarbonate surface with amino groups. We demonstrate the ability of this facile method to serve as a foundation upon which other functionalities may be attached, including anti-fouling coatings and oriented membrane proteins. The use of water as the solvent for the functionalization ensures that solvent induced swelling does not affect the optical or mechanical properties of the polycarbonate.

  6. MIDP-based Realization of a Simple Phone Contact Book

    NASA Astrophysics Data System (ADS)

    Niu, Yan; Xia, Heng; Huan, Lele

    This paper describes the architecture of J2ME and MIDP specification, use the Java language to implement a simple cell phone contact book system, to complete a contact to add, delete, modify, query functions. Different from existing phone contacts, it can run any MIDP-enabled mobile phones, avoid the question of using tool software into and out of phone contact book after user change phone.

  7. Simple, Benign, Aqueous-Based Amination of Polycarbonate Surfaces

    SciTech Connect

    VanDelinder, Virginia; Wheeler, David R.; Small, Leo J.; Brumbach, Michael T.; Spoerke, Erik D.; Henderson, Ian; Bachand, George D.

    2015-03-18

    Here we report a simple, safe, environmentally-friendly aqueous method that uses diamines to functionalize a polycarbonate surface with amino groups. We demonstrate the ability of this facile method to serve as a foundation upon which other functionalities may be attached, including anti-fouling coatings and oriented membrane proteins. The use of water as the solvent for the functionalization ensures that solvent induced swelling does not affect the optical or mechanical properties of the polycarbonate.

  8. Fluorogen-based reporters for fluorescence imaging: a review

    NASA Astrophysics Data System (ADS)

    Jullien, Ludovic; Gautier, Arnaud

    2015-12-01

    Fluorescence bioimaging has recently jumped into a new area of spatiotemporal resolution and sensitivity thanks to synergistic advances in both optical physics and probe/biosensor design. This review focuses on the recent development of genetically encodable fluorescent reporters that bind endogenously present or exogenously applied fluorogenic chromophores (so-called fluorogens) and activate their fluorescence. We highlight the innovative engineering and design that gave rise to these new natural and synthetic fluorescent reporters, and describe some of the emerging applications in imaging and biosensing.

  9. Heavy water: a simple solution to increasing the brightness of fluorescent proteins in super-resolution imaging.

    PubMed

    Ong, Wei Qiang; Citron, Y Rose; Schnitzbauer, Joerg; Kamiyama, Daichi; Huang, Bo

    2015-09-11

    Photoactivatable fluorescent proteins (PA-FPs) are widely used in live single-molecule super-resolution imaging but emit substantially fewer photons than organic dyes do. Herein, we show that in heavy water (D2O) instead of H2O, common PA-FPs emit 26-54% more photons, effectively improving the localization precision in super-resolution imaging. PMID:26226621

  10. Fluorescence Rise Time Measurements for High Temperature Fluorescence-Based Thermometry

    SciTech Connect

    Allison, S.W.

    2005-03-24

    Certain ceramic-like phosphor materials exhibit bright fluorescence with a pronounced temperature dependence over a range which spans the cryogenic to 1700 C, depending on the specific phosphor. To measure temperature, a surface, for instance a turbine blade, is coated with the material. An optical system, sometimes including optical fibers, conveys stimulating light and collects the emission for analysis. Either emission intensity or decay time may indicate temperature. Previously fielded tests have involved surfaces such as blades, vanes, pistons, in-take valves, sheets of galvanneal steel, etc. The fluorescent coatings may be applied to small parts via sputtering methods or to large areas by mixture with inorganic binders. Presented here are results characterizing fluorescence rise times as a means of determining temperature from ambient to 700 C for Y{sub 2}O{sub 3}:Eu.

  11. Fluorescence-based visualization of autophagic activity predicts mouse embryo viability

    NASA Astrophysics Data System (ADS)

    Tsukamoto, Satoshi; Hara, Taichi; Yamamoto, Atsushi; Kito, Seiji; Minami, Naojiro; Kubota, Toshiro; Sato, Ken; Kokubo, Toshiaki

    2014-03-01

    Embryo quality is a critical parameter in assisted reproductive technologies. Although embryo quality can be evaluated morphologically, embryo morphology does not correlate perfectly with embryo viability. To improve this, it is important to understand which molecular mechanisms are involved in embryo quality control. Autophagy is an evolutionarily conserved catabolic process in which cytoplasmic materials sequestered by autophagosomes are degraded in lysosomes. We previously demonstrated that autophagy is highly activated after fertilization and is essential for further embryonic development. Here, we developed a simple fluorescence-based method for visualizing autophagic activity in live mouse embryos. Our method is based on imaging of the fluorescence intensity of GFP-LC3, a versatile marker for autophagy, which is microinjected into the embryos. Using this method, we show that embryonic autophagic activity declines with advancing maternal age, probably due to a decline in the activity of lysosomal hydrolases. We also demonstrate that embryonic autophagic activity is associated with the developmental viability of the embryo. Our results suggest that embryonic autophagic activity can be utilized as a novel indicator of embryo quality.

  12. Synthesis and fluorescence study of 7-azaindole in DNA oligonucleotides replacing a purine base

    PubMed Central

    Wang, Ke; Stringfellow, Sandra; Dong, Shiming; Yu, Hongtao

    2013-01-01

    The fluorescence spectroscopy of 7-azaindole (7aIn) incorporated in DNA oligonucleotides is investigated. Incorporation of 7aIn into DNA oligonucleotides is accomplished through standard solid-phase phosphoramidite chemistry. Fluorescence emission of the 7aIn chromophore shifts slightly to the red (from 386 nm to 388 nm) upon glycosylation at the N ? 1 position, but its relative fluorescence quantum yield increases 23 times, from 0.023 to 0.53. Upon incorporation into DNA, the fluorescence emission of 7aIn is greatly quenched with fluorescence quantum yields of 0.020 and 0.016 in single and double strand DNA, respectively. The fluorescence emission for 7aIn in DNA oligonucleotides shifts to the blue with an emission maximum at 379 nm. Both the strong fluorescence quenching and the blue shift of the emission spectrum signify that 7aIn is stacked with neighboring DNA bases in both single and double strand DNA. As the duplex DNA melts due to temperature increase, the fluorescence of the 7aIn chromophore increases, indicating the transition from the less fluorescent duplex DNA to the more fluorescent single strand DNA. Since this fluorescent 7aIn is a structural analog of purine, its fluorescence property may be utilized as a probe for studying nucleic acid structure and dynamics. PMID:12396042

  13. Synthesis and fluorescence study of 7-azaindole in DNA oligonucleotides replacing a purine base

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Stringfellow, Sandra; Dong, Shiming; Jiao, Yugao; Yu, Hongtao

    2002-10-01

    The fluorescence spectroscopy of 7-azaindole ( 7aIn) incorporated in DNA oligonucleotides is investigated. Incorporation of 7aIn into DNA oligonucleotides is accomplished through standard solid-phase phosphoramidite chemistry. Fluorescence emission of the 7aIn chromophore shifts slightly to the red (from 386 nm to 388 nm) upon glycosylation at the N-1 position, but its relative fluorescence quantum yield increases 23 times, from 0.023 to 0.53. Upon incorporation into DNA, the fluorescence emission of 7aIn is greatly quenched with fluorescence quantum yields of 0.020 and 0.016 in single and double strand DNA, respectively. The fluorescence emission for 7aIn in DNA oligonucleotides shifts to the blue with an emission maximum at 379 nm. Both the strong fluorescence quenching and the blue shift of the emission spectrum signify that 7aIn is stacked with neighboring DNA bases in both single and double strand DNA. As the duplex DNA melts due to temperature increase, the fluorescence of the 7aIn chromophore increases, indicating the transition from the less fluorescent duplex DNA to the more fluorescent single strand DNA. Since this fluorescent 7aIn is a structural analog of purine, its fluorescence property may be utilized as a probe for studying nucleic acid structure and dynamics.

  14. Rapid and quantitative detection of 4(5)-methylimidazole in caramel colours: A novel fluorescent-based immunochromatographic assay.

    PubMed

    Wu, Xinlan; Huang, Minghui; Yu, Shujuan; Kong, Fansheng

    2016-01-01

    A novel fluorescence-based immunochromatographic assay (ICA) for rapid detecting 4(5)-methylimidazole (4-MI) is presented in this study. In our work, the conjugates of fluorescent microspheres (FMs) and 4-MI monoclonal antibody were used as probe for ICA. Under optimal conditions, a standard curve of ICA-based detection of 4-MI was developed, linear detection ranged from 0.50 to 32.0 mg/L. The cross-reactivities were observed less than 3.93% by detecting 6 selected structural analogues of 4-MI. The recoveries of 4-MI in caramels detection were ranged from 82.85% to 102.31%, with the coefficient of variation (n = 3) below 9.06%. Quantitative comparison of the established fluorescence-based ICA with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) analysis of real caramel colour samples indicated a good correlation among the methods. Therefore, our developed fluorescence-based ICA method shows great potential for simple, rapid, sensitive, and cost-effective quantitative detection of 4-MI in food safety control. PMID:26213047

  15. Latest methods of fluorescence-based protein crystal identification

    SciTech Connect

    Meyer, Arne; Betzel, Christian

    2015-01-28

    Fluorescence, whether intrinsic or by using trace fluorescent labeling, can be a powerful aid in macromolecule crystallization. Its use in screening for crystals is discussed here. Successful protein crystallization screening experiments are dependent upon the experimenter being able to identify positive outcomes. The introduction of fluorescence techniques has brought a powerful and versatile tool to the aid of the crystal grower. Trace fluorescent labeling, in which a fluorescent probe is covalently bound to a subpopulation (<0.5%) of the protein, enables the use of visible fluorescence. Alternatively, one can avoid covalent modification and use UV fluorescence, exploiting the intrinsic fluorescent amino acids present in most proteins. By the use of these techniques, crystals that had previously been obscured in the crystallization drop can readily be identified and distinguished from amorphous precipitate or salt crystals. Additionally, lead conditions that may not have been obvious as such under white-light illumination can be identified. In all cases review of the screening plate is considerably accelerated, as the eye can quickly note objects of increased intensity.

  16. A novel fluorescence-based array biosensor: principle and application to DNA hybridization assays.

    PubMed

    Schultz, E; Galland, R; Du Bouëtiez, D; Flahaut, T; Planat-Chrétien, A; Lesbre, F; Hoang, A; Volland, H; Perraut, F

    2008-02-28

    A novel fluorescence-based array biosensor targeted for field applications, such as environmental monitoring, has been developed, and successfully applied to DNA hybridization assays. The purpose was to meet the demand for automated, portable but easy-to-maintain systems allowing continuous flow monitoring of surface reactions. The biosensor presented here can be distinguished from the existing systems by the optical method used, which provides an enhanced simplicity and robustness, and enables a simple maintenance by potentially unskilled personnel. The system is based on a conventional microscope slide which acts both as transducer and biological array sensor. The excited fluorescence is guided by total internal reflection into the slide to the detector which is directly interfaced to the slide. Each region of the sensor array is successively optically interrogated, and the detection of the corresponding fluorescent emission synchronized. A real-time three-analyte analysis is thus feasible without any mechanical scanning movement or optical imaging systems as generally used in the existing instruments. The ability of the biosensor to operate in continuous flow for several tens of hours has been demonstrated. The biosensor has been assessed in terms of stability, and slide-to-slide reproducibility, which is found to be less than 3.7%, thus far below the standard biological reproducibility. DNA hybridization assays were performed to estimate a limit of detection, which was found to be 16 mol/microm(2), and to determine the reaction kinetics associated to the DNA model used. The developed biosensor is thus shown to be able to predict reaction kinetics, and to monitor in real time surface reactions between targets and probes. PMID:18207730

  17. Gold Nanocluster-Assisted Fluorescent Detection for Hydrogen Peroxide and Cholesterol Based on the Inner Filter Effect of Gold Nanoparticles.

    PubMed

    Chang, Heng-Chia; Ho, Ja-an Annie

    2015-10-20

    We developed a simple, sensitive inner filter effect (IFE)-based fluorescent assay for sensing H2O2 and cholesterol. In the process, poly(vinylpyrrolidone)-protected gold nanoparticles (PVP-AuNPs) and fluorescent BSA-protected gold nanoclusters (BSA-AuNCs) were used as an IFE absorber/fluorophore pair. PVP-AuNPs can be a powerful absorber to influence the emission of the fluorophore, BSA-AuNCs, in the IFE-based fluorescent assays. That is due to the high extinction coefficient of AuNPs and the complementary overlap between the surface plasmon resonance (SPR) absorption of PVP-AuNPs and the excitation of BSA-AuNCs. The PVP-Au seeds, produced by directly mixing PVP with HAuCl4, were able to catalyze H2O2 to enlarge AuNPs. The SPR absorption of PVP-AuNPs was enhanced with an increased concentration of H2O2 and, subsequently, induced significant fluorescence quenching of BSA-AuNCs. The IFE-based fluorescent assay enabled the detection of H2O2 and generation of H2O2 in the presence of O2/cholesterol and cholesterol oxidase (ChOx) by the fluorescence response of BSA-AuNCs. The present IFE-based approach can detect H2O2 ranging from 1 to 100 ?M with a detection limit of 0.8 ?M and cholesterol ranging from 1 to 100 ?M with a detection limit of 1.4 ?M. PMID:26379119

  18. Fluorescent glutathione probe based on MnO2-phenol formaldehyde resin nanocomposite.

    PubMed

    Wang, Xudong; Wang, Dan; Guo, Yali; Yang, Chengduan; Liu, Xiaoyu; Iqbal, Anam; Liu, Weisheng; Qin, Wenwu; Yan, Dan; Guo, Huichen

    2016-03-15

    MnO2-phenol formaldehyde resin (MnO2-PFR) nanocomposite is successfully prepared by a simple chemical reduction process. The resultant MnO2-PFR nanocomposite is well characterized. The absorption band of non-fluorescent MnO2 nanosheets overlaps well with the fluorescence emission of PFR nanoparticles. The green fluorescence of PFR in this nanocomposite can be effectively quenched by fluorescence resonance energy transfer from PFR to MnO2. In the presence of glutathione (GSH), the fluorescence of PFR could be recovered due to MnO2 was reduced to Mn(2+) by GSH. The nanocomposite can be use for detecting glutathione in blood serum. PMID:26426853

  19. Carbon dots-based fluorescent probe for "off-on" sensing of Hg(II) and I(.).

    PubMed

    He, Jiangling; Zhang, Haoran; Zou, Jinliang; Liu, Yingliang; Zhuang, Jianle; Xiao, Yong; Lei, Bingfu

    2016-05-15

    Herein, we report a simple, one-step reflux method for synthesis of photoluminescent carbon dots (CDs) using citric acid as the carbon source and diethylenetriamine (DETA) as the surface passivation reagent along with a high quantum yield (82.40%), the fluorescence intensity of the CDs was found to be effectively quenched by Hg(II) ions. Upon addition of I(-) to the CDs/Hg(II) complex dispersion, the fluorescence intensity of the CDs was significantly recovered. Furthermore, we developed an "off-on" fluorescence assay for the detection of I(-) using CDs/Hg(II) as a fluorescence probe. This probe enables the selective detection of Hg(II) with a linear range of 0-80μM and a limit of detection is 0.201µM and a limit of detection about I(-) is 0.234µM with a linear range of 0-70μM. Most importantly, the sensors can be successfully applied to the determination of Hg(II) and I(-) in real lake water and urine of cattles, the "off-on" sensor demonstrates high selectivity, repeatability, stability, which offer this CDs-based "off-on" fluorescent sensor a promising platform for environmental and biological sensing applications. PMID:26748370

  20. Bayesian-based deconvolution fluorescence microscopy using dynamically updated nonstationary expectation estimates

    PubMed Central

    Wong, Alexander; Wang, Xiao Yu; Gorbet, Maud

    2015-01-01

    Fluorescence microscopy is widely used for the study of biological specimens. Deconvolution can significantly improve the resolution and contrast of images produced using fluorescence microscopy; in particular, Bayesian-based methods have become very popular in deconvolution fluorescence microscopy. An ongoing challenge with Bayesian-based methods is in dealing with the presence of noise in low SNR imaging conditions. In this study, we present a Bayesian-based method for performing deconvolution using dynamically updated nonstationary expectation estimates that can improve the fluorescence microscopy image quality in the presence of noise, without explicit use of spatial regularization. PMID:26054051

  1. Feature based registration of fluorescent LSCM imagery using region centroids

    NASA Astrophysics Data System (ADS)

    Lee, Sang-Chul; Bajcsy, Peter

    2005-04-01

    We present a novel semi-automated registration technique for 3D volume reconstruction from fluorescent laser scanning confocal microscope (LSCM) imagery. The developed registration procedure consists of (1) highlighting segmented regions as salient feature candidates, (2) defining two region correspondences by a user, (3) computing a pair of region centroids, as control points for registration, and (4) transforming images according to estimated transformation parameters determined by solving a set of linear equations with input control points. The presented semi-automated method is designed based on our observations that (a) an accurate point selection is much harder for a human than an accurate region (segment) selection, (b) a centroid selection of any region is less accurate by a human than by a computer, and (c) registration based on structural shape of a region rather than on intensity-defined point is more robust to noise and to morphological deformation of features across stacks. We applied the method to image mosaicking and image alignment registration steps and evaluated its performance with 20 human subjects on LSCM images with stained blood vessels. Our experimental evaluation showed significant benefits of automation for 3D volume reconstruction in terms of achieved accuracy, consistency of results and performance time. In addition, the results indicate that the differences between registration accuracy obtained by experts and by novices disappear with an advanced automation while the absolute registration accuracy increases.

  2. Photosystem II Does Not Possess a Simple Excitation Energy Funnel: Time-Resolved Fluorescence Spectroscopy Meets Theory

    PubMed Central

    2013-01-01

    The experimentally obtained time-resolved fluorescence spectra of photosystem II (PS II) core complexes, purified from a thermophilic cyanobacterium Thermosynechococcus vulcanus, at 5180 K are compared with simulations. Dynamic localization effects of excitons are treated implicitly by introducing exciton domains of strongly coupled pigments. Exciton relaxations within a domain and exciton transfers between domains are treated on the basis of Redfield theory and generalized Frster theory, respectively. The excitonic couplings between the pigments are calculated by a quantum chemical/electrostatic method (Poisson-TrEsp). Starting with previously published values, a refined set of site energies of the pigments is obtained through optimization cycles of the fits of stationary optical spectra of PS II. Satisfactorily agreement between the experimental and simulated spectra is obtained for the absorption spectrum including its temperature dependence and the linear dichroism spectrum of PS II core complexes (PS II-CC). Furthermore, the refined site energies well reproduce the temperature dependence of the time-resolved fluorescence spectrum of PS II-CC, which is characterized by the emergence of a 695 nm fluorescence peak upon cooling down to 77 K and the decrease of its relative intensity upon further cooling below 77 K. The blue shift of the fluorescence band upon cooling below 77 K is explained by the existence of two red-shifted chlorophyll pools emitting at around 685 and 695 nm. The former pool is assigned to Chl45 or Chl43 in CP43 (Chl numbering according to the nomenclature of Loll et al. Nature2005, 438, 1040) while the latter is assigned to Chl29 in CP47. The 695 nm emitting chlorophyll is suggested to attract excitations from the peripheral light-harvesting complexes and might also be involved in photoprotection. PMID:23537277

  3. Novel Chalcone-Based Fluorescent Human Histamine H3 Receptor Ligands as Pharmacological Tools

    PubMed Central

    Tomasch, Miriam; Schwed, J. Stephan; Weizel, Lilia; Stark, Holger

    2012-01-01

    Novel fluorescent chalcone-based ligands at human histamine H3 receptors (hH3R) have been designed, synthesized, and characterized. Compounds described are non-imidazole analogs of ciproxifan with a tetralone motif. Tetralones as chemical precursors and related fluorescent chalcones exhibit affinities at hH3R in the same concentration range like the reference antagonist ciproxifan (hH3R pKi value of 7.2). Fluorescence characterization of our novel ligands shows emission maxima about 570?nm for yellow fluorescent chalcones and ?600?nm for the red fluorescent derivatives. Interferences to cellular autofluorescence could be excluded. All synthesized chalcone compounds could be used to visualize hH3R proteins in stably transfected HEK-293 cells using confocal laser scanning fluorescence microscopy. These novel fluorescent ligands possess high potential to be used as pharmacological tools for hH3R visualization in different tissues. PMID:22470321

  4. Neoplasm diagnostics based on fluorescence of polymethine dyes

    NASA Astrophysics Data System (ADS)

    Samtsov, Michael P.; Voropay, Eugene S.; Chalov, Vadim N.; Zhavrid, Edvard A.

    2002-05-01

    Investigated polymethine dye TICS has near IR bands of fluorescence and absorption within the transparency region of biological tissues. It can be detected up to 1.5 cm from the surface of the skin. The intensity of a fluorescence signal of TICS is linear for doses up to 2 mg/kg in both tumor and muscle tissue. The ratio of an intensity of light induced fluorescence in tumor tissue to one in muscle tissue is up to 3.6 for rapidly growing tumors. The retention time of TICS is 7 days in all tissues. TICS can be used in the detection of tumor boundaries and tumor internal structure.

  5. pH sensitivity of FRET reporters based on cyan and yellow fluorescent proteins.

    PubMed

    Betolngar, Dahdjim-Benot; Erard, Marie; Pasquier, Hlne; Bousmah, Yasmina; Diop-Sy, Awa; Guiot, Elvire; Vincent, Pierre; Mrola, Fabienne

    2015-05-01

    It is generally acknowledged that the popular cyan and yellow fluorescent proteins carried by genetically encoded reporters suffer from strong pH sensitivities close to the physiological pH range. We studied the consequences of these pH responses on the intracellular signals of model Frster resonant energy transfer (FRET) tandems and FRET-based reporters of cAMP-dependent protein kinase activity (AKAR) expressed in the cytosol of living BHK cells, while changing the intracellular pH by means of the nigericin ionophore. Although the simultaneous pH sensitivities of the donor and the acceptor may mask each other in some cases, the magnitude of the perturbations can be very significant, as compared to the functional response of the AKAR biosensor. Replacing the CFP donor by the spectrally identical, but pH-insensitive Aquamarine variant (pK1/2 = 3.3) drastically modifies the biosensor pH response and gives access to the acid transition of the yellow acceptor. We developed a simple model of pH-dependent FRET and used it to describe the expected pH-induced changes in fluorescence lifetime and ratiometric signals. This model qualitatively accounts for most of the observations, but reveals a complex behavior of the cytosolic AKAR biosensor at acid pHs, associated to additional FRET contributions. This study underlines the major and complex impact of pH changes on the signal of FRET reporters in the living cell. PMID:25814274

  6. Immunosensor based on fluorescence quenching matrix of the conducting polymer polypyrrole.

    PubMed

    Ramanavicius, A; Ryskevic, N; Oztekin, Y; Kausaite-Minkstimiene, A; Jursenas, S; Baniukevic, J; Kirlyte, J; Bubniene, U; Ramanaviciene, A

    2010-12-01

    In this study, the combination of autofluorescent proteins and fluorescence quenching polymers was shown to be a design which can increase the selectivity and sensitivity of immunosensors. With this objective, the conducting polymer polypyrrole (Ppy) was used as a matrix for immobilization of proteins, which enables biological recognition of the analyte, and as a fluorescence quencher, which increases the selectivity of fluorescence-based detection. In this study, bovine leukemia virus proteins gp51 were immobilized within the Ppy matrix and formed a polymeric layer with affinity for antibodies against protein gp51 (anti-gp51). The anti-gp51 antibodies are present at high levels in the blood serum of cattle infected by bovine leukemia virus. Secondary antibodies labeled with horseradish peroxidase (HRP) were used as specific fluorescent probes for detection of a particular target, because the fluorescence of HRP was readily detectable at the required sensitivity. The Ppy was used as fluorescent background, because its fluorescence was almost undetectable when excited by near UV light at 325 nm. Moreover the Ppy quenched the fluorescence of some fluorescent agents including fluorescein-5(6)-isothiocyanate (fluorescein), rhodamine B, and HRP by almost 100% when these fluorescent agents were adsorbed on the surface of Ppy. It is predicted that Ppy-induced fluorescence quenching could be used in the design of immunosensors to increase selectivity and sensitivity. PMID:20941482

  7. Magnetic quantum tunneling: insights from simple molecule-based magnets.

    PubMed

    Hill, Stephen; Datta, Saiti; Liu, Junjie; Inglis, Ross; Milios, Constantinos J; Feng, Patrick L; Henderson, John J; del Barco, Enrique; Brechin, Euan K; Hendrickson, David N

    2010-05-28

    This perspectives article takes a broad view of the current understanding of magnetic bistability and magnetic quantum tunneling in single-molecule magnets (SMMs), focusing on three families of relatively simple, low-nuclearity transition metal clusters: spin S = 4 Ni(II)(4), Mn(III)(3) (S = 2 and 6) and Mn(III)(6) (S = 4 and 12). The Mn(III) complexes are related by the fact that they contain triangular Mn(III)(3) units in which the exchange may be switched from antiferromagnetic to ferromagnetic without significantly altering the coordination around the Mn(III) centers, thereby leaving the single-ion physics more-or-less unaltered. This allows for a detailed and systematic study of the way in which the individual-ion anisotropies project onto the molecular spin ground state in otherwise identical low- and high-spin molecules, thus providing unique insights into the key factors that control the quantum dynamics of SMMs, namely: (i) the height of the kinetic barrier to magnetization relaxation; and (ii) the transverse interactions that cause tunneling through this barrier. Numerical calculations are supported by an unprecedented experimental data set (17 different compounds), including very detailed spectroscopic information obtained from high-frequency electron paramagnetic resonance and low-temperature hysteresis measurements. Comparisons are made between the giant spin and multi-spin phenomenologies. The giant spin approach assumes the ground state spin, S, to be exact, enabling implementation of simple anisotropy projection techniques. This methodology provides a basic understanding of the concept of anisotropy dilution whereby the cluster anisotropy decreases as the total spin increases, resulting in a barrier that depends weakly on S. This partly explains why the record barrier for a SMM (86 K for Mn(6)) has barely increased in the 15 years since the first studies of Mn(12)-acetate, and why the tiny Mn(3) molecule can have a barrier approaching 60% of this record. Ultimately, the giant spin approach fails to capture all of the key physics, although it works remarkably well for the purely ferromagnetic cases. Nevertheless, diagonalization of the multi-spin Hamiltonian matrix is necessary in order to fully capture the interplay between exchange and local anisotropy, and the resultant spin-state mixing which ultimately gives rise to the tunneling matrix elements in the high symmetry SMMs (ferromagnetic Mn(3) and Ni(4)). The simplicity (low-nuclearity, high-symmetry, weak disorder, etc.) of the molecules highlighted in this study proves to be of crucial importance. Not only that, these simple molecules may be considered among the best SMMs: Mn(6) possesses the record anisotropy barrier, and Mn(3) is the first SMM to exhibit quantum tunneling selection rules that reflect the intrinsic symmetry of the molecule. PMID:20405069

  8. Fluorescence diffuse optical tomography: a wavelet-based model reduction

    NASA Astrophysics Data System (ADS)

    Frassati, Anne; DaSilva, Anabela; Dinten, Jean-Marc; Georges, Didier

    2007-07-01

    Fluorescence diffuse optical tomography is becoming a powerful tool for the investigation of molecular events in small animal studies for new therapeutics developments. Here, the stress is put on the mathematical problem of the tomography, that can be formulated in terms of an estimation of physical parameters appearing as a set of Partial Differential Equations (PDEs). The Finite Element Method has been chosen here to resolve the diffusion equation because it has no restriction considering the geometry or the homogeneity of the system. It is nonetheless well-known to be time and memory consuming, mainly because of the large dimensions of the involved matrices. Our principal objective is to reduce the model in order to speed up the model computation. For that, a new method based on a multiresolution technique is chosen. All the matrices appearing in the discretized version of the PDEs are projected onto an orthonormal wavelet basis, and reduced according to the multiresolution method. With the first order resolution, this compression leads to the reduction of a factor 2x2 of the initial dimension, the inversion of the matrices is approximately 4 times faster. A validation study on a phantom was conducted to evaluate the feasibility of this reduction method.

  9. ALA-based fluorescent diagnosis of malignant oral lesions in the presence of bacterial porphyrin formation

    NASA Astrophysics Data System (ADS)

    Schleier, P.; Berndt, A.; Zinner, K.; Zenk, W.; Dietel, W.; Pfister, W.

    2006-02-01

    The aminolevulinic acid (5-ALA) -based fluorescence diagnosis has been found to be promising for an early detection and demarcation of superficial oral squamous cell carcinomas (OSCC). This method has previously demonstrated high sensitivity, however this clinical trial showed a specificity of approximately 62 %. This specificity was mainly restricted by tumor detection in the oral cavity in the presence of bacteria. After topical ALA application in the mouth of patients with previously diagnosed OSSC, red fluorescent areas were observed which did not correlate to confirm histological findings. Swabs and plaque samples were taken from 44 patients and cultivated microbiologically. Fluorescence was investigated (OMA-system) from 32 different bacteria strains found naturally in the oral cavity. After ALA incubation, 30 of 32 strains were found to synthesize fluorescent porphyrins, mainly Protoporphyrin IX. Also multiple fluorescent spectra were obtained having peak wavelengths of 636 nm and around 618 nm - 620 nm indicating synthesis of different porphyrins, such as the lipophylic Protoporphyrin IX (PpIX) and hydrophylic porphyrins (water soluble porphyrins, wsp). Of the 32 fluorescent bacterial strains, 18 produced wsp, often in combination with PpIX, and 5 produced solely wsp. These results clarify that ALA-based fluorescence diagnosis without consideration or suppression of bacteria fluorescence may lead to false-positive findings. It is necessary to suppress bacteria fluorescence with suitable antiseptics before starting the procedure. In this study, when specific antiseptic pre-treatment was performed bacterial associated fluorescence was significantly reduced.

  10. Fluorescence lifetime-based sensing of sodium by an optode.

    PubMed

    Schwarze, Thomas; Mller, Holger; Ast, Sandra; Steinbrck, Drte; Eidner, Sascha; Geissler, Felix; Kumke, Michael U; Holdt, Hans-Jrgen

    2014-11-25

    We report a 1,2,3-triazol fluoroionophore for detecting Na(+) that shows in vitro enhancement in the Na(+)-induced fluorescence intensity and decay time. The Na(+)-selective molecule 1 was incorporated into a hydrogel as a part of a fiber optical sensor. This sensor allows the direct determination of Na(+) in the range of 1-10 mM by measuring reversible fluorescence decay time changes. PMID:25277847

  11. Determination of flumioxazin residue in food samples through a sensitive fluorescent sensor based on click chemistry.

    PubMed

    Lu, Lijun; Yang, Linlin; Cai, Huijian; Zhang, Lan; Lin, Zhenyu; Guo, Longhua; Qiu, Bin; Chen, Guonan

    2014-11-01

    A sensitive and selective fluorescent sensor for flumioxazin was designed based on the formation of strong fluorescence compound (1,2,3-triazole compounds) via the reaction of the alkynyl group in flumioxazin with 3-azido-7-hydroxycoumarin, a weak-fluorescent compound, through the Cu(+)-catalysed azide-alkyne cycloaddition (CuAAC) reaction. The fluorescence increase factor (represented by F/F0) of the system exhibited a good linear relationship with the concentrations of flumioxazin in the range of 0.25-6.0 ?g/L with a detection limit of 0.18 ?g/L (S/N=3). Also, the proposed fluorescent sensor demonstrated good selectivity for flumioxazin assay even in the presence of high concentration of other pesticides. Based on such high sensitivity and selectivity, the proposed fluorescent sensor has been applied to test the flumioxazin residue in some vegetable and water samples with satisfied results. PMID:24874382

  12. A simple chip free-flow electrophoresis for monosaccharide sensing via supermolecule interaction of boronic acid functionalized quencher and fluorescent dye.

    PubMed

    Yin, Xiao-Yang; Dong, Jing-Yu; Wang, Hou-Yu; Li, Si; Fan, Liu-Yin; Cao, Cheng-Xi

    2013-08-01

    Here, a simple micro free-flow electrophoresis (?FFE) was developed for fluorescence sensing of monosaccharide via supermolecule interaction of synthesized boronic acid functionalized benzyl viologen (?-BBV) and fluorescent dye. The ?FFE contained two open electrode cavities and an ion-exchange membrane was sandwiched between two polymethylmethacrylate plates. The experiments demonstrated the following merits of developed ?FFE: (i) up to 90.5% of voltage efficiency due to high conductivity of ion-exchange membrane; (ii) a strong ability against influence of bubble produced in two electrodes due to open design of electrode cavities; and (iii) reusable and washable separation chamber (45 mm 17 mm 100 ?m, 77 ?L) avoiding the discard of ?FFE due to blockage of solute precipitation in chamber. Remarkably, the ?FFE was first designed for the sensing of monosaccharide via the supermolecule interaction of synthesized ?-BBV, fluorescent dye, and monosaccharide. Under the optimized conditions, the minimum concentration of monosaccharide that could be detected was 1 10(-11) M. Finally, the developed device was used for the detection of 0.3 mM glucose spiked in human urine. All of the results demonstrated the feasibility of monosaccharide detection via the ?FFE. PMID:23712879

  13. Complexation induced fluorescence and acid-base properties of dapoxyl dye with γ-cyclodextrin: a drug-binding application using displacement assays.

    PubMed

    Pal, Kaushik; Mallick, Suman; Koner, Apurba L

    2015-06-28

    Host-guest complexation of dapoxyl sodium sulphonate (DSS), an intramolecular charge transfer dye with water-soluble and non-toxic macrocycle γ-cyclodextrin (γ-CD), has been investigated in a wide pH range. Steady-state absorption, fluorescence and time-resolved fluorescence measurements confirm the positioning of DSS into the hydrophobic cavity of γ-CD. A large fluorescence enhancement ca. 30 times, due to 1 : 2 complex formation and host-assisted guest-protonation have been utilised for developing a method for the utilisation of CD based drug-delivery applications. A simple fluorescence-displacement based approach is implemented at physiological pH for the assessment of binding strength of pharmaceutically useful small drug molecules (ibuprofen, paracetamol, methyl salicylate, salicylic acid, aspirin, and piroxicam) and six important antibiotic drugs (resazurin, thiamphenicol, chloramphenicol, ampicillin, kanamycin, and sorbic acid) with γ-CD. PMID:26028009

  14. Visual fluorescence detection of H2O2 and glucose based on "molecular beacon"-hosted Hoechst dyes.

    PubMed

    Lu, Ling-Fei; Li, Yan-Yun; Zhang, Min; Shi, Guoyue

    2015-05-21

    In this work, a label-free molecular beacon (MB)-like biosensor is designed for the determination of H2O2 and glucose based on the fluorescence regulation of Hoechst dyes hosted by the designed AT-rich single-stranded DNA (ssDNA), in which Hg(2+) and cysteine (Cys) act as activators. The designed AT-rich ssDNA (ATprobe) can be directed to form a hairpin with an Hg(2+)-induced T-Hg(2+)-T complex, which provides a medium for enhancing the fluorescence of Hoechst dyes significantly. On the other hand, Cys can effectively grab Hg(2+) from the T-Hg(2+)-T complex by thiol-Hg(2+) interactions, destructing the hairpin and then switching the Hoechst dyes to the fluorescence "off" state. Combined with these properties, we have demonstrated its application for label-free fluorescence "turn on" detection of H2O2. The sensing mechanism is based on the specific reaction between H2O2 and Cys catalyzed by I(-), the resulting disulfide reverses the Cys-mediated fluorescence decrease of the MB-hosted Hoechst dyes. The approach achieves a low detection limit of 0.1 μM for H2O2. Moreover, this method is further applied to the noninvasive detection of glucose in artificial saliva and urine samples, combining with glucose oxidase (GOx) for the oxidation of glucose and formation of H2O2. Compared to traditional methods, the proposed design is cost-effective, simple to prepare and manipulate without fluorescence labeling or chemical modification. PMID:25868604

  15. Unusual non-fluorescent broad spectrum siderophore activity (SID EGYII) by Pseudomonas aeruginosa strain EGYII DSM 101801 and a new insight towards simple siderophore bioassay.

    PubMed

    Embaby, Amira M; Heshmat, Yasmin; Hussein, Ahmed

    2016-12-01

    Present study highlights an unusual non-fluorescent hydroxamate broad spectrum siderophore (SID EGYII) activity from Pseudomonas aeruginosa strain EGYII DSM 101801, a soil bacterial isolate, along with simple low cost effective siderophore bioassay. Detection of SID EGYII activity qualitatively was proved by masking this activity against Erwinia amylovora strain EGY1 DSM 101800, an indicator strain, in well-cut diffusion assay containing 100 µM FeCl3. SID EGYII activity was expressed quantitatively as arbitrary units [Siderophore arbitrary units (SAU)] 380 SAU/mL against E. amylovora strain EGY1 DSM 101800. Maximal SID EGYII activity was achieved upon growing P. aeruginosa strain EGYII DSM 101801 in PYB broth at 180 rpm for 24 h. SID EGYII displayed a broad spectrum antimicrobial activity against some human pathogens (i.e., Gram-positive bacteria, Gram-negative bacteria and yeasts) and a fireblight plant pathogen. Interestingly, transformants of Escherichia coli JM109 (DE3)pSID/EGYII harboring P. aeruginosa strain EGYII DSM 101801 plasmid demonstrated a perceivable antimicrobial activity against E. amylovora strain EGY1 DSM 101800. The broad spectrum antimicrobial activity of the unusual non-fluorescent SID EGYII would underpin its high potential in targeting bacterial pathogens posing probable threats to human health and agricultural economy. The present simple low cost effective bioassay is a new insight towards an alternative to the expensive cumbersome siderophore Chrome Azurol S assay. PMID:27015845

  16. A simple one-step method to prepare fluorescent carbon dots and their potential application in non-invasive glioma imaging

    NASA Astrophysics Data System (ADS)

    Ruan, Shaobo; Qian, Jun; Shen, Shun; Zhu, Jianhua; Jiang, Xinguo; He, Qin; Gao, Huile

    2014-08-01

    Fluorescent carbon dots (CD) possess impressive potential in bioimaging because of their low photobleaching, absence of optical blinking and good biocompatibility. However, their relatively short excitation/emission wavelengths restrict their application in in vivo imaging. In the present study, a kind of CD was prepared by a simple heat treatment method using glycine as the only precursor. The diameter of CD was lower than 5 nm, and the highest emission wavelength was 500 nm. However, at 600 nm, there was still a relatively strong fluorescent emission, suggesting CD could be used for in vivo imaging. Additionally, several experiments demonstrated that CD possessed good serum stability and low cytotoxicity. In vitro, CD could be taken up into C6 glioma cells in a time- and concentration-dependent manner, with both endosomes and mitochondria involved. In vivo, CD could be used for non-invasive glioma imaging because of its high accumulation in the glioma site of the brain, which was demonstrated by both in vivo imaging and ex vivo tissue imaging. Furthermore, the fluorescent distribution in tissue slices also showed CD distributed in glioma with high intensity, while with a low intensity in normal brain tissue. In conclusion, CD were prepared using a simple method with relatively long excitation and emission wavelengths and could be used for non-invasive glioma imaging.Fluorescent carbon dots (CD) possess impressive potential in bioimaging because of their low photobleaching, absence of optical blinking and good biocompatibility. However, their relatively short excitation/emission wavelengths restrict their application in in vivo imaging. In the present study, a kind of CD was prepared by a simple heat treatment method using glycine as the only precursor. The diameter of CD was lower than 5 nm, and the highest emission wavelength was 500 nm. However, at 600 nm, there was still a relatively strong fluorescent emission, suggesting CD could be used for in vivo imaging. Additionally, several experiments demonstrated that CD possessed good serum stability and low cytotoxicity. In vitro, CD could be taken up into C6 glioma cells in a time- and concentration-dependent manner, with both endosomes and mitochondria involved. In vivo, CD could be used for non-invasive glioma imaging because of its high accumulation in the glioma site of the brain, which was demonstrated by both in vivo imaging and ex vivo tissue imaging. Furthermore, the fluorescent distribution in tissue slices also showed CD distributed in glioma with high intensity, while with a low intensity in normal brain tissue. In conclusion, CD were prepared using a simple method with relatively long excitation and emission wavelengths and could be used for non-invasive glioma imaging. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr02657h

  17. Synthesis and properties of novel base-discriminating fluorescent (BDF) nucleosides.

    PubMed

    Saito, Yoshio; Hanawa, Kazuo; Hayashi, Keigo; Motegi, Kaori; Okaoto, Akimitsu; Saito, Isao

    2005-01-01

    We designed a new type of pyrene-labeled base-discrimination fluorescent (BDF) nucleosides (Py)U, (Py)C, (8Py)A and (MePy)dA, which emitted strong fluorescence only when the bases opposite the BDF base are A, G, T and C, respectively. The DNA probes containing four different BDF bases enable us to distinguish single base alterations by simply mixing with a sample solution of target DNA. PMID:17150679

  18. Two Schiff-base fluorescent sensors for selective sensing of aluminum (III): Experimental and computational studies

    NASA Astrophysics Data System (ADS)

    Qin, Jing-Can; Cheng, Xiao-ying; Fang, Ran; Wang, Ming-fang; Yang, Zheng-yin; Li, Tian-rong; Li, Yong

    2016-01-01

    Two Schiff-base fluorescent sensors have been synthesized, which both can act as fluorescent probes for Al3+, upon addition of Al3+, they exhibit a large fluorescence enhancement which might be attributed to the formation of 1:1 ligand-Al complexes which inhibit photoinduced electron transfer (PET) progress, and that the proposed binding modes of the sensors and Al3+ are identified by theoretical calculations.

  19. Two Schiff-base fluorescent sensors for selective sensing of aluminum (III): Experimental and computational studies.

    PubMed

    Qin, Jing-Can; Cheng, Xiao-Ying; Fang, Ran; Wang, Ming-Fang; Yang, Zheng-Yin; Li, Tian-Rong; Li, Yong

    2016-01-01

    Two Schiff-base fluorescent sensors have been synthesized, which both can act as fluorescent probes for Al(3+), upon addition of Al(3+), they exhibit a large fluorescence enhancement which might be attributed to the formation of 1:1 ligand-Al complexes which inhibit photoinduced electron transfer (PET) progress, and that the proposed binding modes of the sensors and Al(3+) are identified by theoretical calculations. PMID:26232579

  20. Fluorescent PET probes based on perylene-3,4,9,10-tetracarboxylic tetraesters.

    PubMed

    Dubey, Rajeev K; Knorr, Gergely; Westerveld, Nick; Jager, Wolter F

    2016-01-27

    Perylene-3,4,9,10-tetracarboxylic tetraester-based fluorescent PET probes with aniline receptors attached either at the peri- or the bay-positions have been synthesized. By attaching aniline receptors at the bay position, pH-sensitive "light-up" probes, with fluorescence quantum yields ΦF > 0.75 and fluorescent enhancements FE > 500 in ethanol, have been obtained. PMID:26740333

  1. A fluorescent biosensor based on carbon dots-labeled oligodeoxyribonucleotide and graphene oxide for mercury (II) detection.

    PubMed

    Cui, Xin; Zhu, Lei; Wu, Jing; Hou, Yu; Wang, Peiyao; Wang, Zhenni; Yang, Mei

    2015-01-15

    As the newest two members of the carbon materials family, carbon dots (CDs) and graphene oxide (GO) possess many excellent optical properties resulting in a wide range of applications. In this work, we successfully synthesized CDs with a high-quantum-yield, and labeled them on oligodeoxyribonucleotide (ODN). The fluorescence of resultant CDs-labeled oligodeoxyribonucleotide (ODN-CDs) was quenched by GO via fluorescence resonance energy transfer. In the presence of Hg(2+), the fluorescence was recovered by the release of ODN-CDs from GO due to the formation of T-Hg(2+)-T duplex. In the light of this theory, we designed a simple, highly sensitive and selective fluorometric Hg(2+) sensor based on CDs-labeled oligodeoxyribonucleotide and GO without complicated, costly and time-consuming operations. Under the optimal conditions, a linear relationship was obtained between relative fluorescence intensity and the concentration of Hg(2+) in the range of 5-200 nM (R(2)=0.974). The present GO-based sensor system is highly selective toward Hg(2+) over a wide range of metal ions and has a detection limit of 2.6 nM. This method is reliable, and has been successfully applied for the detection of Hg(2+) in practical samples. PMID:25137567

  2. Label-free fluorescent sensor for lead ion detection based on lead(II)-stabilized G-quadruplex formation.

    PubMed

    Zhan, Shenshan; Wu, Yuangen; Luo, Yanfang; Liu, Le; He, Lan; Xing, Haibo; Zhou, Pei

    2014-10-01

    A label-free fluorescent DNA sensor for the detection of lead ions (Pb(2+)) based on lead(II)-stabilized G-quadruplex formation is proposed in this article. A guanine (G)-rich oligonucleotide, T30695, was used as a recognition probe, and a DNA intercalator, SYBR Green I (SG), was used as a signal reporter. In the absence of Pb(2+), the SG intercalated with the single-stranded random-coil T30695 and emitted strong fluorescence. While in the presence of Pb(2+), the random-coil T30695 would fold into a G-quadruplex structure and the SG could barely show weak fluorescence, and the fluorescence intensity was inversely proportional to the involving amount of Pb(2+). Based on this, a selective lead ion sensor with a limit of detection of 3.79 ppb (parts per billion) and a detection range from 0 to 600 ppb was constructed. Because detection for real samples was also demonstrated to be reliable, this simple, low-cost, sensitive, and selective sensor holds good potential for Pb(2+) detection in real environmental samples. PMID:24486320

  3. Simple membrane-based model of the Min oscillator

    NASA Astrophysics Data System (ADS)

    Petrášek, Zdeněk; Schwille, Petra

    2015-04-01

    Min proteins in E. coli bacteria organize into a dynamic pattern oscillating between the two cell poles. This process identifies the middle of the cell and enables symmetric cell division. In an experimental model system consisting of a flat membrane with effectively infinite supply of proteins and energy source, the Min proteins assemble into travelling waves. Here we propose a simple one-dimensional model of the Min dynamics that, unlike the existing models, reproduces the sharp decrease of Min concentration when the majority of protein detaches from the membrane, and even the narrow MinE maximum immediately preceding the detachment. The proposed model thus provides a possible mechanism for the formation of the MinE ring known from cells. The model is restricted to one dimension, with protein interactions described by chemical kinetics allowing at most bimolecular reactions, and explicitly considering only three, membrane-bound, species. The bulk solution above the membrane is approximated as being well-mixed, with constant concentrations of all species. Unlike other models, our proposal does not require autocatalytic binding of MinD to the membrane. Instead, it is assumed that two MinE molecules are necessary to induce the dissociation of the MinD dimer and its subsequent detachment from the membrane. We investigate which reaction schemes lead to unstable homogeneous steady states and limit cycle oscillations, and how diffusion affects their stability. The suggested model qualitatively describes the shape of the Min waves observed on flat membranes, and agrees with the experimental dependence of the wave period on the MinE concentration. These results highlight the importance of MinE presence on the membrane without being bound to MinD, and of the reactions of Min proteins on the membrane.

  4. A sensitive quantum dots-based "OFF-ON" fluorescent sensor for ruthenium anticancer drugs and ctDNA.

    PubMed

    Huang, Shan; Zhu, Fawei; Qiu, Hangna; Xiao, Qi; Zhou, Quan; Su, Wei; Hu, Baoqing

    2014-05-01

    In this contribution, a simple and sensitive fluorescent sensor for the determination of both the three ruthenium anticancer drugs (1 to 3) and calf thymus DNA (ctDNA) was established based on the CdTe quantum dots (QDs) fluorescence "OFF-ON" mode. Under the experimental conditions, the fluorescence of CdTe QDs can be effectively quenched by ruthenium anticancer drugs because of the surface binding of these drugs on CdTe QDs and the subsequent photoinduced electron transfer (PET) process from CdTe QDs to ruthenium anticancer drugs, which render the system into fluorescence "OFF" status. The system can then be "ON" after the addition of ctDNA which brought the restoration of CdTe QDs fluorescence intensity, since ruthenium anticancer drugs broke away from the surface of CdTe QDs and inserted into double helix structure of ctDNA. The fluorescence quenching effect of the CdTe QDs-ruthenium anticancer drugs systems was mainly concentration dependent, which could be used to detect three ruthenium anticancer drugs. The limits of detection were 5.5 10(-8) M for ruthenium anticancer drug 1, 7.0 10(-8) M for ruthenium anticancer drug 2, and 7.9 10(-8) M for ruthenium anticancer drug 3, respectively. The relative restored fluorescence intensity was directly proportional to the concentration of ctDNA in the range of 1.0 10(-8) M ? 3.0 10(-7) M, with a correlation coefficient (R) of 0.9983 and a limit of detection of 1.1 10(-9) M. The relative standard deviation (RSD) for 1.5 10(-7) M ctDNA was 1.5% (n = 5). There was almost no interference to some common chemical compounds, nucleotides, amino acids, and proteins. The proposed method was applied to the determination of ctDNA in three synthetic samples with satisfactory results. The possible reaction mechanism of CdTe QDs fluorescence "OFF-ON" was further investigated. This simple and sensitive approach possessed some potential applications in the investigation of interaction between drug molecules and DNA. PMID:24657609

  5. Recent progress in design of protein-based fluorescent biosensors and their cellular applications.

    PubMed

    Tamura, Tomonori; Hamachi, Itaru

    2014-12-19

    Protein-based fluorescent biosensors have emerged as key bioanalytical tools to visualize and quantify a wide range of biological substances and events in vitro, in cells, and even in vivo. On the basis of the construction method, the protein-based fluorescent biosensors can be principally classified into two classes: (1) genetically encoded fluorescent biosensors harnessing fluorescent proteins (FPs) and (2) semisynthetic biosensors comprised of protein scaffolds and synthetic fluorophores. Recent advances in protein engineering and chemical biology not only allowed the further optimization of conventional biosensors but also facilitated the creation of novel biosensors based on unique strategies. In this review, we survey the recent studies in the development and improvement of protein-based fluorescent biosensors and highlight the successful applications to live cell and in vivo imaging. Furthermore, we provide perspectives on possible future directions of the technique. PMID:25317665

  6. Fluorescent nanodiamonds for ultrasensitive detection

    NASA Astrophysics Data System (ADS)

    Kimball, Joseph; Shumilov, Dmytro; Maliwa, Badri; Zerda, T. W.; Rout, Bibhu; Fudala, Rafal; Raut, Sangram; Gryczynski, Ignacy; Simanek, Eric; Borejdo, Julian; Rich, Ryan; Akopova, Irina; Gryczynski, Zygmunt

    2014-03-01

    Fluorescent nanodiamonds (NDs) are new and emerging nanomaterials that have potential to be used as fluorescence imaging agents and also as a highly versatile platform for the controlled functionalization and delivery of a wide spectrum of therapeutic agents. We will utilize two experimental methods, TIRF, a relatively simple method based on total internal reflection fluorescence and SPRF, fluorescence enhanced by resonance coupling with surface plasmons. We estimate that the SPRF method will be 100 times sensitive than currently available similar detectors based on detectors. The ultimate goal of this research is to develop microarray platforms that could be used for sensitive, fast and inexpensive gene sequencing and protein detection.

  7. Simple hobby computer-based off-gas analysis system

    SciTech Connect

    Forrest, E.H.; Jansen, N.B.; Flickinger, M.C.; Tsao, G.T.

    1981-02-01

    An Apple II computer has been adapted to monitor fermentation offgas in laboratory and pilot scale fermentors. It can calculate oxygen uptake rates, carbon dioxide evolution rates, respiratory quotient as well as initiating recalibration procedures. In this report the computer-based off-gas analysis system is described.

  8. Reaction analysis on Yb(3+) and DNA based on quantum dots: The design of a fluorescent reversible off-on mode.

    PubMed

    Wang, Linlin; Song, Jing; Liu, Shaopu; Hao, Chenxia; Kuang, Nianxi; He, Youqiu

    2015-11-01

    Even though various strategies have reported for DNA detection, development of a simple, time-saving and specific fluorescent sensing platform still remains a desired goal. In this work, a quantum dots (QDs) based fluorescent reversible "off-on" mode was developed for sensitively recognition of herring sperm DNA (hsDNA). Firstly, in the "turn off" stage, the fluorescence of glutathione (GSH) capped CdTe QDs could be effectively quenched by ytterbium ion (Yb(3+)) was due to the occurrence of the electron transfer between Yb(3+) and the photoexcited QDs. And then, in the following "turn on" stage, with the effective binding reaction of Yb(3+) to hsDNA, the fluorescence intensity of GSH-capped CdTe QDs enhanced. Under the optimal conditions, the linear range of fluorescence versus the concentration of hsDNA was 0.010-12 ?g/mL, and the detection limit was 3.033 ng/mL. In addition, the reaction mechanism among GSH-capped CdTe QDs, Yb(3+) and hsDNA were investigated by fluorescence spectroscopy, UV-vis spectrophotometry, fluorescence lifetime measurement and viscosity measurements. This analytical fluorescent reversible "off-on" pattern offered a way with good sensitivity and selectivity for the detection of hsDNA. PMID:26164248

  9. Gold Nanoparticles-Coated SU-8 for Sensitive Fluorescence-Based Detections of DNA

    PubMed Central

    Cao, Cuong; Birtwell, Sam W.; Hgberg, Jonas; Morgan, Hywel; Wolff, Anders; Bang, Dang Duong

    2012-01-01

    SU-8 epoxy-based negative photoresist has been extensively employed as a structural material for fabrication of numerous biological microelectro-mechanical systems (Bio-MEMS) or lab-on-a-chip (LOC) devices. However, SU-8 has a high autofluorescence level that limits sensitivity of microdevices that use fluorescence as the predominant detection workhorse. Here, we show that deposition of a thin gold nanoparticles layer onto the SU-8 surface significantly reduces the autofluorescence of the coated SU-8 surface by as much as 81% compared to bare SU-8. Furthermore, DNA probes can easily be immobilized on the Au surface with high thermal stability. These improvements enabled sensitive DNA detection by simple DNA hybridization down to 1 nM (a two orders of magnitude improvement) or by solid-phase PCR with sub-picomolar sensitivity. The approach is simple and easy to perform, making it suitable for various Bio-MEMs and LOC devices that use SU-8 as a structural material.

  10. Dissolved oxygen sensing based on fluorescence quenching of ceria nanoparticles

    NASA Astrophysics Data System (ADS)

    Shehata, Nader; Meehan, Kathleen; Leber, Donald

    2012-10-01

    The development of oxygen sensors has positively impacted the fields of medical science, bioengineering, environmental monitoring, solar cells, industrial process control, and a number of military applications. Fluorescent quenching sensors have an inherent high sensitivity, chemical selectivity, and stability when compared to other types of sensors. While cerium oxide thin films have been used to monitor oxygen in the gas phase, the potential of cerium oxide (ceria) nanoparticles as the active material in sensor for oxygen gas has only recently been investigated. Ceria nanoparticles are one of the most unique nanomaterials that are being studied today due to the diffusion and reactivity of its oxygen vacancies, which contributes to its high oxygen storage capability. The reactivity of the oxygen vacancies, which is also related to conversion of cerium ion from the Ce+4 to Ce+3 state, affects the fluorescence properties of the ceria nanoparticles. Our research demonstrates that the ceria nanoparticles (~7 nm in diameter) have application as a fluorescence quenching sensor to measure dissolved oxygen in water. We have found a strong inverse correlation between the amplitude of the fluorescence emission (?excitation = 430 nm and ?peak = 520 nm) and the dissolved oxygen concentration between 5 - 13 mg/L. The Stern-Volmer constant, which is an indication of the sensitivity of gas sensing is 184 M-1 for the ceria nanoparticles. The results show that ceria nanoparticles can be used in an improved, robust fluorescence sensor for dissolved oxygen in a liquid medium.

  11. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy

    PubMed Central

    Tyson, Adam L.; Hilton, Stephen T.; Andreae, Laura C.

    2015-01-01

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  12. Rapid, simple and inexpensive production of custom 3D printed equipment for large-volume fluorescence microscopy.

    PubMed

    Tyson, Adam L; Hilton, Stephen T; Andreae, Laura C

    2015-10-30

    The cost of 3D printing has reduced dramatically over the last few years and is now within reach of many scientific laboratories. This work presents an example of how 3D printing can be applied to the development of custom laboratory equipment that is specifically adapted for use with the novel brain tissue clearing technique, CLARITY. A simple, freely available online software tool was used, along with consumer-grade equipment, to produce a brain slicing chamber and a combined antibody staining and imaging chamber. Using standard 3D printers we were able to produce research-grade parts in an iterative manner at a fraction of the cost of commercial equipment. 3D printing provides a reproducible, flexible, simple and cost-effective method for researchers to produce the equipment needed to quickly adopt new methods. PMID:25797056

  13. A novel, simple and efficient dye laser with low amplified spontaneous emission background for analytical fluorescence and ionization spectroscopy

    SciTech Connect

    Matveev, Oleg I.; Omenetto, Nicolo'

    1995-04-01

    A new, simple, compact and efficient, grazing- incidence type of dye laser is suggested which has a low level of Amplified Spontaneous Emission. By using a Coumarin dye (LD 5000) pumped with a 20 mJ XeCl excimer laser, and a diffraction grating with 3000 grooves/mm, an efficiency of 11%, a spectral bandwidth of 0.6 cm{sup -1} and a tuning range from 458 to 517 nm have been obtained.

  14. Designation of rapid detection system for chlorophyll fluorescence parameters based on LED irradiation

    NASA Astrophysics Data System (ADS)

    Li, Zhengming; Ji, Jianwei; Xu, Minghu

    2013-03-01

    Adopting high-power light-emitting diode (LED) as excitation light source, the study designed a rapid detection system for fluorescence parameters based on MINIPAM. The system uses a microcomputer as the core of the programmable power supply to provide constant current drive of the LED array, and the LED array as a fluorescence excitation light source produces light photochemical system needed. It also uses MINIPAM to detect the fluorescence, analyzing the fluorescence parameters of the mathematical model, studying the plant photosystem& light response curve. The System is of great significance in the evaluation of chlorophyll photosynthesis ability and the plant physiological stress response and the appropriate mechanism.

  15. Fluorescence lifetime imaging from time resolved measurements using a shape-based approach.

    PubMed

    Alvarez, Diego; Medina, Paúl; Moscoso, Miguel

    2009-05-25

    We present a novel fluorescent tomography algorithm to estimate the spatial distribution of fluorophores and the fluorescence lifetimes from surface time resolved measurements. The algorithm is a hybridization of the level set technique for recovering the distributions of distinct fluorescent markers with a gradient method for estimating their lifetimes. This imaging method offers several advantages compared to more traditional pixel-based techniques as, for example, well defined boundaries and a better resolution of the images. The numerical experiments show that our imaging method gives rise to accurate reconstructions in the presence of data noise and fluorescence background even for complicated fluorophore distributions in several-centimiter-thick biological tissue. PMID:19466134

  16. Cytometric sorting based on the fluorescence lifetime of spectrally overlapping signals

    PubMed Central

    Cao, Ruofan; Pankayatselvan, Varayini; Houston, Jessica P.

    2013-01-01

    Flow cytometry is a well-established and powerful high-throughput fluorescence measurement tool that also allows for the sorting and enrichment of subpopulations of cells expressing unique fluorescence signatures. Owing to the reliance on intensity-only signals, flow cytometry sorters cannot easily discriminate between fluorophores that spectrally overlap. In this paper we demonstrate a new method of cell sorting using a fluorescence lifetime-dependent methodology. This approach, referred to herein as phase-filtered cell sorting (PFCS), permits sorting based on the average fluorescence lifetime of a fluorophore by separating fluorescence signals from species that emit differing average fluorescence lifetimes. Using lifetime-dependent hardware, cells and microspheres labeled with fluorophores were sorted with purities up to 90%. PFCS is a practical approach for separating populations of cells that are stained with spectrally overlapping fluorophores or that have interfering autofluorescence signals. PMID:23787669

  17. A turn-on fluorescent probe for hypochlorous acid based on the oxidation of diphenyl telluride.

    PubMed

    Venkatesan, Parthiban; Wu, Shu-Pao

    2015-02-21

    A fluorescent probe HCTe was developed for rapid detection of hypochlorous acid based on the specific HOCl-promoted oxidation of diphenyl telluride. The reaction is accompanied by an 82-fold increase in the fluorescence quantum yield (from 0.009 to 0.75). The fluorescence turn-on mechanism is achieved by the suppression of photoinduced electron transfer (PET) from the diphenyl telluride group to BODIPY. The fluorescence intensity of the reaction between HOCl and HCTe is linear in the HOCl concentration range of 1 to 10 ?M with a detection limit of 41.3 nM (S/N = 3). In addition, confocal fluorescence microscopy imaging using RAW264.7 macrophages demonstrated that HCTe could be an efficient fluorescent probe for HOCl detection in living cells. PMID:25580477

  18. Development of and Clinical Experience with a Simple Device for Performing Intraoperative Fluorescein Fluorescence Cerebral Angiography: Technical Notes

    PubMed Central

    ICHIKAWA, Tsuyoshi; SUZUKI, Kyouichi; WATANABE, Yoichi; SATO, Taku; SAKUMA, Jun; SAITO, Kiyoshi

    2016-01-01

    To perform intraoperative fluorescence angiography (FAG) under a microscope without an integrated FAG function with reasonable cost and sufficient quality for evaluation, we made a small and easy to use device for fluorescein FAG (FAG filter). We investigated the practical use of this FAG filter during aneurysm surgery, revascularization surgery, and brain tumor surgery. The FAG filter consists of two types of filters: an excitatory filter and a barrier filter. The excitatory filter excludes all wavelengths except for blue light and the barrier filter passes long waves except for blue light. By adding this FAG filter to a microscope without an integrated FAG function, light from the microscope illuminating the surgical field becomes blue, which is blocked by the barrier filter. We put the FAG filter on the objective lens of the operating microscope correctly and fluorescein sodium was injected intravenously or intra-arterially. Fluorescence (green light) from vessels in the surgical field and the dyed tumor were clearly observed through the microscope and recorded by a memory device. This method was easy and could be performed in a short time (about 10 seconds). Blood flow of small vessels deep in the surgical field could be observed. Blood flow stagnation could be evaluated. However, images from this method were inferior to those obtained by currently commercially available microscopes with an integrated FAG function. In brain tumor surgery, a stained tumor on the brain surface could be observed using this method. FAG could be performed with a microscope without an integrated FAG function easily with only this FAG filter. PMID:26597335

  19. Development of and Clinical Experience with a Simple Device for Performing Intraoperative Fluorescein Fluorescence Cerebral Angiography: Technical Notes.

    PubMed

    Ichikawa, Tsuyoshi; Suzuki, Kyouichi; Watanabe, Yoichi; Sato, Taku; Sakuma, Jun; Saito, Kiyoshi

    2016-03-15

    To perform intraoperative fluorescence angiography (FAG) under a microscope without an integrated FAG function with reasonable cost and sufficient quality for evaluation, we made a small and easy to use device for fluorescein FAG (FAG filter). We investigated the practical use of this FAG filter during aneurysm surgery, revascularization surgery, and brain tumor surgery. The FAG filter consists of two types of filters: an excitatory filter and a barrier filter. The excitatory filter excludes all wavelengths except for blue light and the barrier filter passes long waves except for blue light. By adding this FAG filter to a microscope without an integrated FAG function, light from the microscope illuminating the surgical field becomes blue, which is blocked by the barrier filter. We put the FAG filter on the objective lens of the operating microscope correctly and fluorescein sodium was injected intravenously or intra-arterially. Fluorescence (green light) from vessels in the surgical field and the dyed tumor were clearly observed through the microscope and recorded by a memory device. This method was easy and could be performed in a short time (about 10 seconds). Blood flow of small vessels deep in the surgical field could be observed. Blood flow stagnation could be evaluated. However, images from this method were inferior to those obtained by currently commercially available microscopes with an integrated FAG function. In brain tumor surgery, a stained tumor on the brain surface could be observed using this method. FAG could be performed with a microscope without an integrated FAG function easily with only this FAG filter. PMID:26597335

  20. Fluorescent sensing of anions with acridinedione based neutral PET chemosensor

    NASA Astrophysics Data System (ADS)

    Thiagarajan, Viruthachalam; Ramamurthy, Perumal

    2007-07-01

    Newly synthesised fluorescent chemosensor ADDTU contains the thiourea receptor connected to the acridinedione (ADD) fluorophore via a covalent bond, giving rise to a fluorophore-receptor motif. In this fluorescent chemosensor, the anion recognition takes place at the receptor site which result in the concomitant changes in the photophysical properties of a ADD fluorophore by modulation of photoinduced electron transfer (PET) process. The binding ability of these sensor with the anions F -, Cl -, Br -, I -, HSO 4-, ClO 4-, AcO -, H 2PO 4- and BF 4- (as their tetrabutylammounium salts) in acetonitrile were investigated using UV-vis, steady state and time-resolved emission techniques. ADDTU system allows for the selective fluorescent sensing of AcO -, H 2PO 4- and F - over other anions in acetonitrile.

  1. Array-based sensing of purine derivatives with fluorescent dyes.

    PubMed

    Kstereli, Ziya; Severin, Kay

    2015-09-21

    Natural and synthetic purine derivatives such as caffeine, theophylline, 6-mercaptopurine and 8-chlorotheophylline are important drugs. Due to the structural similarity of these compounds, it is intrinsically difficult to prepare chemosensors for their selective optical detection. Here, we describe a sensor array which can be used to differentiate pharmacologically important purine derivatives with good accuracy. The array is composed of four polysufonated fluorescent dyes, all of which can bind purines via?-stacking interactions. The complexation of the analytes results in partial quenching of the fluorescence. The fluorescence response of the four dyes provides a characteristic signal pattern, enabling the identification of thirteen purine derivatives at low millimolar concentration. Furthermore, it is possible to use the array for obtaining information about the quantity and purity of purine samples. PMID:26223489

  2. A simple data base for identification of risk profiles

    SciTech Connect

    Munganahalli, D.

    1996-12-31

    Sedco Forex is a drilling contractor that operates approximately 80 rigs on land and offshore worldwide. The HSE management system developed by Sedco Forex is an effort to prevent accidents and minimize losses. An integral part of the HSE management system is establishing risk profiles and thereby minimizing risk and reducing loss exposures. Risk profiles are established based on accident reports, potential accident reports and other risk identification reports (RIR) like the Du Pont STOP system. A rig could fill in as many as 30 accident reports, 30 potential accident reports and 500 STOP cards each year. Statistics are important for an HSE management system, since they are indicators of success or failure of HSE systems. It is however difficult to establish risk profiles based on statistical information, unless tools are available at the rig site to aid with the analysis. Risk profiles are then used to identify important areas in the operation that may require specific attention to minimize the loss exposure. Programs to address the loss exposure can then be identified and implemented with either a local or corporate approach. In January 1995, Sedco Forex implemented a uniform HSE Database on all the rigs worldwide. In one year companywide, the HSE database would contain information on approximately 500 accident and potential accident reports, and 10,000 STOP cards. This paper demonstrates the salient features of the database and describes how it has helped in establishing key risk profiles. It also shows a recent example of how risk profiles have been established at the corporate level and used to identify the key contributing factors to hands and finger injuries. Based on this information, a campaign was launched to minimize the frequency of occurrence and associated loss attributed to hands and fingers accidents.

  3. Modulation of a solid-state reversible fluorescent photoswitching based on a controllable photochromic pyrazolones

    SciTech Connect

    Liu, Hu; Guo, Jixi; Jia, Dianzeng; Guo, Mingxi; Le, Fuhe; Liu, Lang; Wu, Dongling; Li, Feng

    2014-08-15

    A novel solid-state reversible fluorescence photoswitching system (FPS) based on photochromism of photochromic pyrazolones has been developed by employing phosphor Sr{sub 2}P{sub 2}O{sub 7} co-doped with europium ion and chlorine ion (Sr{sub 2}P{sub 2}O{sub 7}–EC) and 1,3-diphenyl-4-(3-chlorobenzal)-5-hydroxypyrazole-4-phenylsemicarbazone (1a) as the fluorescence dye and the photochromic compound, respectively. With carefully selected components, the absorption band of the keto-form photochromic pyrazolones well overlaps with the emission peak of Sr{sub 2}P{sub 2}O{sub 7}–EC. The fluorescence emission intensity of Sr{sub 2}P{sub 2}O{sub 7}–EC is efficiently modulated by the photoisomerization of 1a with controlling the exposure time in the solid state. The fluorescence photoswitching system displayed high fluorescence quenching efficiency and remarkable fatigue resistance. It can be repeated 7 cycles without observable the changes of emission intensity. A fluorescence quenching efficiency can be achieved with a reversible colour change from white to yellow. - Graphical abstract: A novel fluorescence photoswitching system based on doping inorganic fluorescence dye into photochromic pyrazolones was constructed successfully. Its fluorescence emission could be efficiently modulated by the photoisomerization of pyrazolones. - Highlights: • A solid-state fluorescence photoswitching material was prepared. • Photoswitching is due to energy transfer between pyrazolone and fluorescence dye. • It exhibits excellent fluorescence contrast and fatigue resistance in the solid state.

  4. A Fluorescent Sensor for Dinitrobenzoic Acid Based on a Cyanuric Acid and Xanthene Skeleton

    PubMed Central

    Muiz, Francisco M.; Simn, Luis; Sez, Silvia; Raposo, Csar; Alczar, Victoria; Morn, Joaqun R.

    2008-01-01

    A new fluorescent sensor based on a dimethylxanthene skeleton has been synthesized. Because of its oxyanion hole structure, this receptor includes a suitable cavity for the association of carboxylic acids. The receptors fluorescence is quenched upon addition of dinitrobenzoic acid.

  5. A chromenoquinoline-based fluorescent off-on thiol probe for bioimaging.

    PubMed

    Kand, Dnyaneshwar; Kalle, Arunasree Marasanapalli; Varma, Sreejith Jayasree; Talukdar, Pinaki

    2012-03-11

    A new chromenoquinoline-based fluorescent off-on thiol probe 2 is reported. In aqueous buffer solutions at physiological pH, the probe exhibited 223-fold enhancement in fluorescence intensity by a Michael addition of cysteine to the maleimide appended to a chromenoquinoline. Cell permeability and live cell imaging of thiols are also demonstrated. PMID:22301487

  6. SIL-based confocal fluorescence microscope for investigating individual nanostructures

    NASA Astrophysics Data System (ADS)

    Krajnik, Bartosz; Schulte, Tim; Piątkowski, Dawid; Czechowski, Nikodem; Hofmann, Eckhard; Mackowski, Sebastian

    2011-04-01

    We developed a fluorescence confocal microscope equipped with a hemispherical solid immersion lens (SIL) and apply it to study the optical properties of light-harvesting complexes. We demonstrate that the collection efficiency of the SIL-equipped microscope is significantly improved, as is the spatial resolution, which reaches 600 nm. This experimental setup is suitable for detailed studies of physical phenomena in hybrid nanostructures. In particular, we compare the results of fluorescence intensity measurements for a light-harvesting peridinin-chlorophyll-protein (PCP) complex with and without the SIL.

  7. Fluorescence-Based Sensor for Monitoring Activation of Lunar Dust

    NASA Technical Reports Server (NTRS)

    Wallace, William T.; Jeevarajan, Antony S.

    2012-01-01

    This sensor unit is designed to determine the level of activation of lunar dust or simulant particles using a fluorescent technique. Activation of the surface of a lunar soil sample (for instance, through grinding) should produce a freshly fractured surface. When these reactive surfaces interact with oxygen and water, they produce hydroxyl radicals. These radicals will react with a terephthalate diluted in the aqueous medium to form 2-hydroxyterephthalate. The fluorescence produced by 2-hydroxyterephthalate provides qualitative proof of the activation of the sample. Using a calibration curve produced by synthesized 2-hydroxyterephthalate, the amount of hydroxyl radicals produced as a function of sample concentration can also be determined.

  8. Fluorescence molecular tomographic image reconstruction based on reduced measurement data

    NASA Astrophysics Data System (ADS)

    Zou, Wei; Wang, Jiajun; Feng, David Dagan; Fang, Erxi

    2015-07-01

    The analysis of fluorescence molecular tomography is important for medical diagnosis and treatment. Although the quality of reconstructed results can be improved with the increasing number of measurement data, the scale of the matrices involved in the reconstruction of fluorescence molecular tomography will also become larger, which may slow down the reconstruction process. A new method is proposed where measurement data are reduced according to the rows of the Jacobian matrix and the projection residual error. To further accelerate the reconstruction process, the global inverse problem is solved with level-by-level Schur complement decomposition. Simulation results demonstrate that the speed of the reconstruction process can be improved with the proposed algorithm.

  9. Quantitative Detection Method of Hydroxyapatite Nanoparticles Based on Eu(3+) Fluorescent Labeling in Vitro and in Vivo.

    PubMed

    Xie, Yunfei; Perera, Thalagalage Shalika Harshani; Li, Fang; Han, Yingchao; Yin, Meizhen

    2015-11-01

    One major challenge for application of hydroxyapatite nanoparticles (nHAP) in nanomedicine is the quantitative detection method. Herein, we exploited one quantitative detection method for nHAP based on the Eu(3+) fluorescent labeling via a simple chemical coprecipitation method. The trace amount of nHAP in cells and tissues can be quantitatively detected on the basis of the fluorescent quantitative determination of Eu(3+) ions in nHAP crystal lattice. The lowest concentration of Eu(3+) ions that can be quantitatively detected is 0.5 nM using DELFIA enhancement solution. This methodology can be broadly applicable for studying the tissue distribution and metabolization of nHAP in vivo. PMID:26495748

  10. Simple communication using a SSVEP-based BCI

    NASA Astrophysics Data System (ADS)

    Sanchez, Guillermo; Diez, Pablo F.; Avila, Enrique; Laciar Leber, Eric

    2011-12-01

    Majority of Brain-Computer Interface (BCI) for communication purposes are speller, i.e., the user has to select letter by letter. In this work, is proposed a different approach where the user can select words from a word set designed in order to answer a wide range of questions. The word selection process is commanded by a Steady-state visual evoked potential (SSVEP) based-BCI that allows selecting a word in an average time of 26 s with accuracies of 92% on average. This BCI is focus in the first stages on rehabilitation or even in first moments of some diseases (such as stroke), when the person is eager to communicate with family and doctors.

  11. Microfluidics-Based Selection of Red-Fluorescent Proteins with Decreased Rates of Photobleaching

    PubMed Central

    Dean, Kevin M.; Lubbeck, Jennifer L.; Davis, Lloyd M.; Regmi, Chola K.; Chapagain, Prem P.; Gerstman, Bernard S.; Jimenez, Ralph; Palmer, Amy E.

    2014-01-01

    Fluorescent proteins offer exceptional labeling specificity in living cells and organisms. Unfortunately, their photophysical properties remain far from ideal for long-term imaging of low-abundance cellular constituents, in large part because of their poor photostability. Despite widespread engineering efforts, improving the photostability of fluorescent proteins remains challenging due to lack of appropriate high-throughput selection methods. Here, we use molecular dynamics guided mutagenesis in conjunction with a recently developed microfluidic-based platform, which sorts cells based on their fluorescence photostability, to identify red fluorescent proteins with decreased photobleaching from a HeLa cell-based library. The identified mutant, named Kriek, has 2.5- and 4-fold higher photostability than its progenitor, mCherry, under widefield and confocal illumination, respectively. Furthermore, the results provide insight into mechanisms for enhancing photostability and their connections with other photophysical processes, thereby providing direction for ongoing development of fluorescent proteins with improved single-molecule and low-copy imaging capabilities. Insight, innovation, integration Fluorescent proteins enable imaging in situ, throughout the visible spectrum, with superb molecular specificity and single-molecule sensitivity. Unfortunately, when compared to leading small-molecule fluorophores (e.g., Cy3), fluorescent proteins, suffer from accelerated photobleaching and poor integrated photon output. This results from a lack of appropriate high-throughput methods for improving the photostability of fluorescent proteins, as well as a poor molecular understanding of fluorescent protein photobleaching. Here, we report the first application of a recently developed microfluidic cell-sorter to identify fluorescent proteins from a mCherry-derived library with improved photostability. The results provide insight into fluorescent protein photophysics, greatly accelerate identification of improved mutants, and can be applied to both genetically encoded and small-molecule fluorophores. PMID:25477249

  12. Neural Bases of Categorization of Simple Speech and Nonspeech Sounds

    PubMed Central

    Husain, Fatima T.; Fromm, Stephen J.; Pursley, Randall H.; Hosey, Lara A.; Braun, Allen R.; Horwitz, Barry

    2016-01-01

    Categorization is fundamental to our perception and understanding of the environment. However, little is known about the neural bases underlying the categorization of sounds. Using human functional magnetic resonance imaging (fMRI) we compared the brain responses to a category discrimination task with an auditory discrimination task using identical sets of sounds. Our stimuli differed along two dimensions: a speech–nonspeech dimension and a fast–slow temporal dynamics dimension. All stimuli activated regions in the primary and nonprimary auditory cortices in the temporal cortex and in the parietal and frontal cortices for the two tasks. When comparing the activation patterns for the category discrimination task to those for the auditory discrimination task, the results show that a core group of regions beyond the auditory cortices, including inferior and middle frontal gyri, dorsomedial frontal gyrus, and intraparietal sulcus, were preferentially activated for familiar speech categories and for novel nonspeech categories. These regions have been shown to play a role in working memory tasks by a number of studies. Additionally, the categorization of nonspeech sounds activated left middle frontal gyrus and right parietal cortex to a greater extent than did the categorization of speech sounds. Processing the temporal aspects of the stimuli had a greater impact on the left lateralization of the categorization network than did other factors, particularly in the inferior frontal gyrus, suggesting that there is no inherent left hemisphere advantage in the categorical processing of speech stimuli, or for the categorization task itself. PMID:16281285

  13. A base-modified PNA-graphene oxide platform as a turn-on fluorescence sensor for the detection of human telomeric repeats

    NASA Astrophysics Data System (ADS)

    Sabale, Pramod M.; George, Jerrin Thomas; Srivatsan, Seergazhi G.

    2014-08-01

    Given the biological and therapeutic significance of telomeres and other G-quadruplex forming sequences in human genome, it is highly desirable to develop simple methods to study these structures, which can also be implemented in screening formats for the discovery of G-quadruplex binders. The majority of telomere detection methods developed so far are laborious and use elaborate assay and instrumental setups, and hence, are not amenable to discovery platforms. Here, we describe the development of a simple homogeneous fluorescence turn-on method, which uses a unique combination of an environment-sensitive fluorescent nucleobase analogue, the superior base pairing property of PNA, and DNA-binding and fluorescence quenching properties of graphene oxide, to detect human telomeric DNA repeats of varying lengths. Our results demonstrate that this method, which does not involve a rigorous assay setup, would provide new opportunities to study G-quadruplex structures.Given the biological and therapeutic significance of telomeres and other G-quadruplex forming sequences in human genome, it is highly desirable to develop simple methods to study these structures, which can also be implemented in screening formats for the discovery of G-quadruplex binders. The majority of telomere detection methods developed so far are laborious and use elaborate assay and instrumental setups, and hence, are not amenable to discovery platforms. Here, we describe the development of a simple homogeneous fluorescence turn-on method, which uses a unique combination of an environment-sensitive fluorescent nucleobase analogue, the superior base pairing property of PNA, and DNA-binding and fluorescence quenching properties of graphene oxide, to detect human telomeric DNA repeats of varying lengths. Our results demonstrate that this method, which does not involve a rigorous assay setup, would provide new opportunities to study G-quadruplex structures. Electronic supplementary information (ESI) available. Figures, tables, experimental procedures and NMR spectra. See DOI: 10.1039/c4nr00878b

  14. Integrated microspectrometer for fluorescence based analysis in a microfluidic format.

    PubMed

    Hu, Zhixiong; Glidle, Andrew; Ironside, Charles N; Sorel, Marc; Strain, Michael J; Cooper, Jon; Yin, Huabing

    2012-08-21

    We have demonstrated a monolithic integrated arrayed waveguide grating (AWG) microspectrometer microfluidic platform capable of fluorescence spectroscopic analysis. The microspectrometer in this proof of concept study has a small (1 cm 1 cm) footprint and 8 output channels centred on different wavelengths. We show that the signals from the output channels detected on a camera chip can be used to recreate the complete fluorescence spectrum of an analyte. By making fluorescence measurements of (i) mixed quantum dot solutions, (ii) an organic fluorophore (Cy5) and (iii) the propidium iodide (PI)-DNA assay, we illustrate the unique advantages of the AWG platform for simultaneous, quantitative multiplex detection and its capability to detect small spectroscopic shifts. Although the current system is designed for fluorescence spectroscopic analysis, in principle, it can be implemented for other types of analysis, such as Raman spectroscopy. Fabricated using established semiconductor industry methods, this miniaturised platform holds great potential to create a handheld, low cost biosensor with versatile detection capability. PMID:22648688

  15. Development of a Green Fluorescent Protein-Based Laboratory Curriculum

    ERIC Educational Resources Information Center

    Larkin, Patrick D.; Hartberg, Yasha

    2005-01-01

    A laboratory curriculum has been designed for an undergraduate biochemistry course that focuses on the investigation of the green fluorescent protein (GFP). The sequence of procedures extends from analysis of the DNA sequence through PCR amplification, recombinant plasmid DNA synthesis, bacterial transformation, expression, isolation, and

  16. Fluorescence based spectral assessment of pork meat freshness

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Development of sensitive, nondestructive measurement methods for meat freshness is necessary to ensure safe distribution of meat products in the continually growing meat market. Fluorescence spectral technology has been shown to be a promising measurement method for quality and safety evaluation of ...

  17. Development of a Green Fluorescent Protein-Based Laboratory Curriculum

    ERIC Educational Resources Information Center

    Larkin, Patrick D.; Hartberg, Yasha

    2005-01-01

    A laboratory curriculum has been designed for an undergraduate biochemistry course that focuses on the investigation of the green fluorescent protein (GFP). The sequence of procedures extends from analysis of the DNA sequence through PCR amplification, recombinant plasmid DNA synthesis, bacterial transformation, expression, isolation, and…

  18. DNA origami-based standards for quantitative fluorescence microscopy.

    PubMed

    Schmied, Jrgen J; Raab, Mario; Forthmann, Carsten; Pibiri, Enrico; Wnsch, Bettina; Dammeyer, Thorben; Tinnefeld, Philip

    2014-01-01

    Validating and testing a fluorescence microscope or a microscopy method requires defined samples that can be used as standards. DNA origami is a new tool that provides a framework to place defined numbers of small molecules such as fluorescent dyes or proteins in a programmed geometry with nanometer precision. The flexibility and versatility in the design of DNA origami microscopy standards makes them ideally suited for the broad variety of emerging super-resolution microscopy methods. As DNA origami structures are durable and portable, they can become a universally available specimen to check the everyday functionality of a microscope. The standards are immobilized on a glass slide, and they can be imaged without further preparation and can be stored for up to 6 months. We describe a detailed protocol for the design, production and use of DNA origami microscopy standards, and we introduce a DNA origami rectangle, bundles and a nanopillar as fluorescent nanoscopic rulers. The protocol provides procedures for the design and realization of fluorescent marks on DNA origami structures, their production and purification, quality control, handling, immobilization, measurement and data analysis. The procedure can be completed in 1-2 d. PMID:24833175

  19. Simple plant-based design strategies for volatile organic pollutants

    SciTech Connect

    Narayanan, M.; Erickson, L.E.; Davis, L.C.

    1999-12-31

    Vegetation which enhances in-situ biodegradation of organic compounds can play a key role in the bioremediation of such contaminants in polluted soils and groundwater. Plants may act directly on some contaminants by degrading them, but their main effect is to enhance microbial populations in the thizosphere. Microbially mediated transformations are thus indirectly facilitated by root exudates which nourish the indigenous microorganisms. Plants may also be viewed as a solar driven pump-and-treat system which can contain a plume and reduce the spread of contaminated water. Laboratory investigations carried out in a growth chamber with alfalfa plants provide evidence for the (microbially mediated) biodegradation of organic compounds such as toluene, phenol and TCE. Alfalfa plants tolerate concentrations of these organics in contaminated water up to 100 mg/L. They facilitate transfer of the contaminants from the saturated to the vadose zone. For volatile organic compounds such as TCE, vegetation provides a controlled release of compounds and hence assures dilution of the TCE evapotranspired into the atmosphere from contaminated soils. Using a range of calculated plausible scenarios, it is shown that intermedia transfer caused by volatilization associated with plants is most unlikely to lead to exceedance of standards for gas phase contamination, for most volatile contaminants. Possible action level exceedances might occur with highly toxic substances including vinyl chloride and carbon tetrachloride, if they re present in ground water at levels above kilogram amounts in a single plume of a few hectares, and released by vigorously growing plants under hot dry conditions. Information needed for the calculation and design of plant-based bioremediation systems for typical sites is discussed in this paper.

  20. Simple adaptive sparse representation based classification schemes for EEG based brain-computer interface applications.

    PubMed

    Shin, Younghak; Lee, Seungchan; Ahn, Minkyu; Cho, Hohyun; Jun, Sung Chan; Lee, Heung-No

    2015-11-01

    One of the main problems related to electroencephalogram (EEG) based brain-computer interface (BCI) systems is the non-stationarity of the underlying EEG signals. This results in the deterioration of the classification performance during experimental sessions. Therefore, adaptive classification techniques are required for EEG based BCI applications. In this paper, we propose simple adaptive sparse representation based classification (SRC) schemes. Supervised and unsupervised dictionary update techniques for new test data and a dictionary modification method by using the incoherence measure of the training data are investigated. The proposed methods are very simple and additional computation for the re-training of the classifier is not needed. The proposed adaptive SRC schemes are evaluated using two BCI experimental datasets. The proposed methods are assessed by comparing classification results with the conventional SRC and other adaptive classification methods. On the basis of the results, we find that the proposed adaptive schemes show relatively improved classification accuracy as compared to conventional methods without requiring additional computation. PMID:26378500

  1. Application of fluorescence-based semi-automated allelotyping to the molecular characterization of tumors

    SciTech Connect

    Jedlicka, A.E.; DiSilvestre, D.; Holroyd, K.J.

    1994-09-01

    In cancer genetics, identifying loss of heterozygosity (LOH) defines candidate regions which warrant further analyses to determine the presence of tumor suppressor genes. In addition, demonstrating LOH has potential utility for improving the pathologic classification of tumors. Molecular methods that improve the efficiency and accuracy of LOH studies will be helpful in both clinical and research applications. Here we demonstrate a fluorescence-based semi-automated alleotyping method for studies of LOH in cancer, using gliomas as an example. Gliomas are tumors arising from neuroglia, the supporting tissue intermingled with essential elements of the brain and spinal cord. Since this method utilizes PCR-based highly polymorphic simple sequence repeat markers, it is suitable for small and archival tumor specimens. We collected tumor tissue from a variety of gliomas, and DNA was extracted. White blood cells from the same individuals served as a source of {open_quotes}control{close_quotes} DNA. We PCR amplified markers from tumor and genomic DNA to detect molecular alterations in six people. Simultaneous analysis of 14 loci near gene candidates on chromosomes 5, 7, 9, 10, 11, and 22, were evaluated. Strikingly, in most cases there was allelic loss in brain tumor compared to genomic DNA for at least one of these loci. In addition, alleles of lesser intensity were also shown at a few loci of the tumor DNA, suggesting possible genetic instability. We conclude from these data that fluorescent semi-automated allelotyping is a quantitative and efficient process for determining and analyzing LOH in gliomas, and possibly other tumors. These methods will facilitate the identification of candidate loci critical in the development and progression of tumors.

  2. Applicability of fluorescence-based sensors to the determination of kinetic parameters for O? in oxygenases.

    PubMed

    Di Russo, Natali V; Bruner, Steven D; Roitberg, Adrian E

    2015-04-15

    Optical methods for O2 determination based on dynamic fluorescence quenching have been applied to measure oxygen uptake rates in cell culture and to determine intracellular oxygen levels. Here we demonstrate the applicability of fluorescence-based probes in determining kinetic parameters for O2 using as an example catalysis by a cofactor-independent oxygenase (DpgC). Fluorescence-based sensors provide a direct assessment of enzyme-catalyzed O2 consumption using commercially available, low-cost instrumentation that is easily customizable and, thus, constitutes a convenient alternative to the widely used Clark-type electrode, especially in cases where chemical interference is expected to be problematic. PMID:25637681

  3. Thiazole Orange Dimers in DNA: Fluorescent Base Substitutions with Hybridization Readout.

    PubMed

    Berndl, Sina; Dimitrov, Stoichko D; Menacher, Florian; Fiebig, Torsten; Wagenknecht, Hans-Achim

    2016-02-01

    By using (S)-2-amino-1,3-propanediol as a linker, thiazole orange (TO) was incorporated in a dimeric form into DNA. The green fluorescence (λ=530 nm) of the intrastrand TO dimer is quenched, whereas the interstrand TO dimer shows a characteristic redshifted orange emission (λ=585 nm). Steady-state optical spectroscopic methods reveal that the TO dimer fluorescence is independent of the sequential base contexts. Time-resolved pump-probe measurements and excitation spectra reveal the coexistence of conformations, including mainly stacked TO dimers and partially unstacked ones, which yield exciton and excimer contributions to the fluorescence, respectively. The helicity of the DNA framework distorts the excitonic coupling. In particular, the interstrand TO dimer could be regarded as an excitonically interacting base pair with fluorescence readout for DNA hybridization. Finally, the use of this fluorescent readout was representatively demonstrated in molecular beacons. PMID:26773846

  4. Simple method of determination of copper, mercury and lead in potable water with preliminary pre-concentration by total reflection X-ray fluorescence spectrometry

    NASA Astrophysics Data System (ADS)

    Ho?y?ska, B.; Ostachowicz, B.; W?grzynek, D.

    1996-06-01

    Total reflection X-ray fluorescence spectrometry and chemical pre-concentration procedures have been applied for the analysis of trace concentrations of copper, mercury, and lead in drinking water samples. A simple total reflection module has been used in X-ray measurements. The elements under investigation were pre-concentrated by complexation using a mixture of carbamates followed by solvent extraction with methyl isobutyl ketone. The preconcentration procedure was tested with the use of twice-distilled water samples and samples of mineral and tap water spiked with known additions of copper, mercury, and lead. The obtained recovery and precision values are presented. The minimum detection limits for the determination of these elements in mineral and tap water samples were found to be 40 ng l -1, 60 ng l -1, and 60 ng l -1, respectively.

  5. Fluorescence switch of dye-infiltrated SiO2 inverse opal based on acid-base vapors or light

    NASA Astrophysics Data System (ADS)

    Zhang, Y. Q.; Wang, J. X.; Shang, Y. L.; Song, Y. L.; Jiang, L.

    2011-03-01

    The acid-base vapors/light double responsive dye-infiltrated SiO2 inverse opal photonic crystals (PCs) were fabricated by sacrificial template method and a subsequent infiltration of spiropyran derivative dye molecules. The fluorescence of ring-open dye molecules infiltrated in PCs can be switched on/off based on different fluorescence properties of spiropyran dye under stimuli of acid-base vapors or light, when PCs with suitable stopband were selected. The fluorescence switch behavior based on PCs has potential applications in data storage, color displays, chemical and biological sensors.

  6. Convenient and selective off-on detection nitric oxide in solution and thin film with quinoline based fluorescence sensor

    NASA Astrophysics Data System (ADS)

    Yu, Miao; Wang, Wei; Zhang, Ning

    Quinoline based fluorescence sensor (1) was synthesized and characterized with mass spectra (MS), 1H nuclear magnetic resonance (1H NMR) spectrometer, elemental analyses, and infrared (IR) spectra. Following fluorescence experiments demonstrate 1 can coordinate with copper ions, and lead to fluorescence completely quenched. The 1-copper complex was used as a turn-on fluorescence biosensor to convenient and highly effective detect nitric oxide (NO) over other radicals in solution and PCL-based thin film. The finding would enable the quinoline based fluorescence probe to be an off-on convenient NO fluorescence probe.

  7. Exciton energy transfer-based quantum dot fluorescence sensing array: "chemical noses" for discrimination of different nucleobases.

    PubMed

    Liu, Jianbo; Li, Gui; Yang, Xiaohai; Wang, Kemin; Li, Li; Liu, Wei; Shi, Xing; Guo, Yali

    2015-01-20

    A novel exciton energy transfer-based fluorescence sensing array for the discrimination of different nucleobases was developed through target nucleobase-triggered self-assembly of quantum dots (QDs). Four QD nanoprobes with different ligand receptors, including mercaptoethylamine, N-acetyl-l-cysteine, 2-dimethyl-aminethanethiol, and thioglycolic acid, were created to detect and identify nucleobase targets. These QDs served as both selective recognition scaffolds and signal transduction elements for a biomolecule target. The extent of particle assembly, induced by the analyte-triggered self-assembly of QDs, led to an exciton energy transfer effect between interparticles that gave a readily detectable fluorescence quenching and distinct fluorescence response patterns. These patterns are characteristic for each nucleobase and can be quantitatively differentiated by linear discriminate analysis. Furthermore, a fingerprint-based barcode was established to conveniently discriminate the nucleobases. This pattern sensing was successfully used to identify nucleobase samples at unknown concentrations and five rare bases. In this "chemical noses" strategy, the robust characteristics of QD nanoprobes, coupled with the diversity of surface functionality that can be readily obtained using nanoparticles, provides a simple and label-free biosensing approach that shows great promise for biomedical applications. PMID:25495103

  8. A graphitic carbon nitride based fluorescence resonance energy transfer detection of riboflavin.

    PubMed

    Han, Jing; Zou, Hong Yan; Gao, Ming Xuan; Huang, Cheng Zhi

    2016-02-01

    Fluorescence resonance energy transfer (FRET), which occurs between two luminescent chromophores, can greatly improve the selectivity and sensitivity of a fluorescent assay when a ratiometric signaling with the fluorescence enhancement of the acceptor at the expense of the donor is adopted. In this study, a fluorescence ratiometric detection (FRD) of riboflavin (RF) has been made based on FRET, as the strong overlap occurred between the emission spectrum of graphitic carbon nitride (g-C3N4) and absorption spectrum of RF, in which g-C3N4 acts as the energy donor and RF as the energy acceptor. With increasing concentration of RF, the fluorescence intensity of g-C3N4 emission at 444 nm decreased and the fluorescence peak at 523 nm for RF increased regularly, making the fluorescence intensity ratio of 523 nm to 444 nm linearly dependent on the concentration of RF in the range from 0.4 μM to 10 μM, giving a limit of the detection of 170 nM. This method can be used to quantify RF in complex systems such as milk and drink, showing that the novel FRET-based fluorescence ratiometric detection can enable an attractive assay platform for analytes of interest. PMID:26653450

  9. A Simple and Sensitive HPLC Method for Fluorescence Quantitation of Doxorubicin in Micro-volume Plasma: Applications to Pharmacokinetic Studies in Rats

    PubMed Central

    Daeihamed, Marjan; Haeri, Azadeh; Dadashzadeh, Simin

    2015-01-01

    A validated HPLC method was developed to determine the doxorubicin concentration in a small volume of rat plasma (60 µL) with convenient fluorescence detection. Sample preparation includes a simple one-step liquid-liquid extraction using a minimum amount of organic solvent, with extraction recovery more than 95%. The analysis was accomplished using PerfectSil C18 column maintained at 35 °C and a mobile phase consisted of acetonitrile and water (32:68, v/v; pH=2.6). The flow-rate was kept at 1 mL/min and the column effluent was monitored with a fluorescence detector at an excitation and emission wavelength of 470 and 555 nm, respectively. The detection limit was 5 ng/mL. No analytical interference was observed from endogenous components in the rat plasma. This method was feasibly applied to the pharmacokinetic study of 5 mg/Kg of doxorubicin after the intravenous administration to rats. PMID:26185503

  10. Fluorescence-based video profile beam diagnostics: Theory and experience

    SciTech Connect

    Sandoval, D.; Gilpatrick, D.; Shinas, M.; Garcia, R.; Yuan, V.; Zander, M.

    1994-05-01

    Inelastic collisions between accelerated particles and residual gas in the accelerator vessel can cause the residual gas to fluoresce. The gas fluorescence intensity is proportional to the current density of the particle beam. This process provides the foundation for a video diagnostic system to measure the profile and position of accelerated particle beams. This, in fact, has proven to be a useful diagnostic at several installations. This paper describes the light production process resulting from beam -- residual gas interactions and gives formulas for estimating the beam radiance for various conditions. Ground Test Accelerator (GTA) radiance calculations will be used as an example. In addition, measurement experiences with the GTA video diagnostics system will be discussed.

  11. Spectrally resolved fluorescence correlation spectroscopy based on global analysis.

    PubMed

    Previte, Michael J R; Pelet, Serge; Kim, Ki Hean; Buehler, Christoph; So, Peter T C

    2008-05-01

    Multicolor fluorescence correlation spectroscopy has been recently developed to study chemical interactions of multiple chemical species labeled with spectrally distinct fluorophores. In the presence of spectral overlap, there exists a lower detectability limit for reaction products with multicolor fluorophores. In addition, the ability to separate bound product from reactants allows thermodynamic properties such as dissociation constants to be measured for chemical reactions. In this report, we utilize a spectrally resolved two-photon microscope with single-photon counting sensitivity to acquire spectral and temporal information from multiple chemical species. Further, we have developed a global fitting analysis algorithm that simultaneously analyzes all distinct auto- and cross-correlation functions from 15 independent spectral channels. We have demonstrated that the global analysis approach allows the concentration and diffusion coefficients of fluorescent particles to be resolved despite the presence of overlapping emission spectra. PMID:18351754

  12. Fluorescence-based video profile beam diagnostics: Theory and experience

    SciTech Connect

    Sandoval, D.P.; Garcia, R.C.; Gilpatrick, J.D.; Shinas, M.A.; Wright, R.; Yuan, V.; Zander, M.E. )

    1994-10-10

    Inelastic collisions between accelerated particles and residual gas in the accelerator vessel can cause the residual gas to fluoresce. The gas fluorescence intensity is proportional to the current density of the particle beam. This process provides the foundation for a video diagnostics system to measure the profile and position of accelerated particle beams. This, in fact, has proven to be a useful diagnostic at several installations. This paper describes the light production process resulting from beam-residual gas interactions and gives formulas for estimating the beam radiance for various conditions. Ground Test Accelerator (GTA) radiance calculations will be used as an example. In addition, measurement experiences with the GTA video diagnostics system will be discussed.

  13. Development of an aptasensor based on a fluorescent particles-modified aptamer for ochratoxin A detection.

    PubMed

    Hayat, Akhtar; Mishra, Rupesh K; Catanante, Gaelle; Marty, Jean Louis

    2015-10-01

    The presented work reports a generic fluorescent aptasensing design employing carboxy-modified fluorescent particles as a signal-generating probe and magnetic particles as a solid separation support. Carboxy-modified fluorescent particles were used to modify the aptamer and act as a signal-generating probe. Magnetic beads were used as an immobilization surface to perform the function of a solid separation support. As a proof of concept, the assay was used to detect ochratoxin A (OTA). Fluorescent detection based on the displacement and competition format was performed, and the obtained results were compared. The competition-based assays were characterized with improved analytical characteristics as compared to those based on the displacement principle. The competitive fluorescent assays showed a high sensitivity where the detection limit and IC50 were 0.005 and 7.2 nM respectively. The aptasensing platform was finally demonstrated for the detection of OTA in a beer sample. However, this is a generic approach that can be very easily extended to other matrixes to determine OTA. Additionally, the proposed concept of fluorescent particles as a signal-generating probe in combination with magnetic particles can also be integrated to other fluorescent-based affinity assays. PMID:26277188

  14. When Simple Harmonic Motion is not That Simple: Managing Epistemological Complexity by Using Computer-based Representations

    NASA Astrophysics Data System (ADS)

    Parnafes, Orit

    2010-12-01

    Many real-world phenomena, even "simple" physical phenomena such as natural harmonic motion, are complex in the sense that they require coordinating multiple subtle foci of attention to get the required information when experiencing them. Moreover, for students to develop sound understanding of a concept or a phenomenon, they need to learn to get the same type of information across different contexts and situations (diSessa and Sherin 1998; diSessa and Wagner 2005). Rather than simplifying complex situations, or creating a linear instructional sequence in which students move from one context to another, this paper demonstrates the use of computer-based representations to facilitate developing understanding of complex physical phenomena. The data is collected from 8 studies in which pairs of students are engaged in an exploratory activity, trying to understand the dynamic behavior of a simulation and, at the same time, to attribute meaning to it in terms of the physical phenomenon it represents. The analysis focuses on three episodes. The first two episodes demonstrate the epistemological complexity involved in attempting to make sense of natural harmonic oscillation. A third episode demonstrates the process by which students develop understanding in this complex perceptual and conceptual territory, through the mediation (Vygotsky 1978) of computer-based representations designed to facilitate understanding in this topic.

  15. Selective recognition of Ni2+ ion based on fluorescence enhancement chemosensor

    NASA Astrophysics Data System (ADS)

    Ganjali, M. R.; Hosseini, M.; Motalebi, M.; Sedaghat, M.; Mizani, F.; Faridbod, F.; Norouzi, P.

    2015-04-01

    A new enhancing fluorescent chemosensor was introduced for selective and sensitive determination of nickel ions based on 2-(1-H-benzo[d]imidazol-2yl)-N-phenyl hydrazine carbothioamide (L). L has an intrinsic fluorescent emission which enhances in presence of nickel ions in CH3CN/H2O (70:30, v/v) solution. The fluorescence enhancement of L is attributed to a 1:1 complex formation between L and Ni2+ ion which has been used for selective detection of Ni2+ ion. At the optimum conditions, the fluorescence intensity of L at 352 nm enhances linearly by the concentration of nickel ion from 1.6 10-5 to 1.6 10-7 M and detection limit of 7.9 10-8 M. The new fluorescent probe exhibited high selectivity to Ni2+ ion over the other common mono, di-and trivalent cations.

  16. Fluorescent nanogel based on four-arm PEG-PCL copolymer with porphyrin core for bioimaging.

    PubMed

    Dong, Xia; Wei, Chang; Lu, Li; Liu, Tianjun; Lv, Feng

    2016-04-01

    Four-arm PEG-PCL copolymer with porphyrin core (POR-PEG-PCL) exhibits beneficial fluorescence ability in vivo. To further develop an application of thermosensitive porphyrin hydrogel based on four-arm PEG-PCL copolymer as a drug carrier, a POR-PEG-PCL nanogel was tracked and located to tumor tissue with porphyrin as a fluorescence tag via intravenous injection. The structure and function of the nanogel were evaluated by TEM, DLS, H-NMR, UV-vis and fluorescence spectra. The fluorescent nanogel was monitored by an in vivo imaging system with hepatoma tumor-bearing mice. Good biocompatibility and safety in vitro and in vivo show that the POR-PEG-PCL nanogel is a potential drug carrier that targets tumor tissues with fluorescence bioimaging. PMID:26838843

  17. A new boronic acid fluorescent sensor based on fluorene for monosaccharides at physiological pH

    NASA Astrophysics Data System (ADS)

    Hosseinzadeh, Rahman; Mohadjerani, Maryam; Pooryousef, Mona; Eslami, Abbas; Emami, Saeed

    2015-06-01

    Fluorescent boronic acids are very useful fluorescent sensor for detection of biologically important saccharides. Herein we synthesized a new fluorene-based fluorescent boronic acid that shows significant fluorescence changes upon addition of saccharides at physiological pH. Upon addition of fructose, sorbitol, glucose, galactose, ribose, and maltose at different concentration to the solution of 7-(dimethylamino)-9,9-dimethyl-9H-fluoren-2-yl-2-boronic acid (7-DMAFBA, 1), significant decreases in fluorescent intensity were observed. It was found that this boronic acid has high affinity (Ka = 3582.88 M-1) and selectivity for fructose over glucose at pH = 7.4. The sensor 1 showed a linear response toward D-fructose in the concentrations ranging from 2.5 10-5 to 4 10-4 mol L-1 with the detection limit of 1.3 10-5 mol L-1.

  18. Angular shaping of fluorescence from synthetic opal-based photonic crystal.

    PubMed

    Boiko, Vitalii; Dovbeshko, Galyna; Dolgov, Leonid; Kiisk, Valter; Sildos, Ilmo; Loot, Ardi; Gorelik, Vladimir

    2015-01-01

    Spectral, angular, and temporal distributions of fluorescence as well as specular reflection were investigated for silica-based artificial opals. Periodic arrangement of nanosized silica globules in the opal causes a specific dip in the defect-related fluorescence spectra and a peak in the reflectance spectrum. The spectral position of the dip coincides with the photonic stop band. The latter is dependent on the size of silica globules and the angle of observation. The spectral shape and intensity of defect-related fluorescence can be controlled by variation of detection angle. Fluorescence intensity increases up to two times at the edges of the spectral dip. Partial photobleaching of fluorescence was observed. Photonic origin of the observed effects is discussed. PMID:25852393

  19. Angular shaping of fluorescence from synthetic opal-based photonic crystal

    NASA Astrophysics Data System (ADS)

    Boiko, Vitalii; Dovbeshko, Galyna; Dolgov, Leonid; Kiisk, Valter; Sildos, Ilmo; Loot, Ardi; Gorelik, Vladimir

    2015-02-01

    Spectral, angular, and temporal distributions of fluorescence as well as specular reflection were investigated for silica-based artificial opals. Periodic arrangement of nanosized silica globules in the opal causes a specific dip in the defect-related fluorescence spectra and a peak in the reflectance spectrum. The spectral position of the dip coincides with the photonic stop band. The latter is dependent on the size of silica globules and the angle of observation. The spectral shape and intensity of defect-related fluorescence can be controlled by variation of detection angle. Fluorescence intensity increases up to two times at the edges of the spectral dip. Partial photobleaching of fluorescence was observed. Photonic origin of the observed effects is discussed.

  20. [Photosynthetic Parameters Inversion Algorithm Study Based on Chlorophyll Fluorescence Induction Kinetics Curve].

    PubMed

    Qiu, Xiao-han; Zhang, Yu-jun; Yin, Gao-fang; Shi, Chao-yi; Yu, Xiao-ya; Zhao, Nan-jing; Liu, Wen-qing

    2015-08-01

    The fast chlorophyll fluorescence induction curve contains rich information of photosynthesis. It can reflect various information of vegetation, such as, the survival status, the pathological condition and the physiology trends under the stress state. Through the acquisition of algae fluorescence and induced optical signal, the fast phase of chlorophyll fluorescence kinetics curve was fitted. Based on least square fitting method, we introduced adaptive minimum error approaching method for fast multivariate nonlinear regression fitting toward chlorophyll fluorescence kinetics curve. We realized Fo (fixedfluorescent), Fm (maximum fluorescence yield), σPSII (PSII functional absorption cross section) details parameters inversion and the photosynthetic parameters inversion of Chlorella pyrenoidosa. And we also studied physiological variation of Chlorella pyrenoidosa under the stress of Cu(2+). PMID:26672292

  1. A rapid fluorescence-based assay for classification of iNKT cell activating glycolipids.

    PubMed

    Arora, Pooja; Venkataswamy, Manjunatha M; Baena, Andres; Bricard, Gabriel; Li, Qian; Veerapen, Natacha; Ndonye, Rachel; Park, Jeong Ju; Lee, Ji Hyung; Seo, Kyung-Chang; Howell, Amy R; Chang, Young-Tae; Illarionov, Petr A; Besra, Gurdyal S; Chung, Sung-Kee; Porcelli, Steven A

    2011-04-13

    Structural variants of ?-galactosylceramide (?GC) that activate invariant natural killer T cells (iNKT cells) are being developed as potential immunomodulatory agents for a variety of applications. Identification of specific forms of these glycolipids that bias responses to favor production of proinflammatory vs anti-inflammatory cytokines is central to current efforts, but this goal has been hampered by the lack of in vitro screening assays that reliably predict the in vivo biological activity of these compounds. Here we describe a fluorescence-based assay to identify functionally distinct ?GC analogues. Our assay is based on recent findings showing that presentation of glycolipid antigens by CD1d molecules localized to plasma membrane detergent-resistant microdomains (lipid rafts) is correlated with induction of interferon-? secretion and Th1-biased cytokine responses. Using an assay that measures lipid raft residency of CD1d molecules loaded with ?GC, we screened a library of ?200 synthetic ?GC analogues and identified 19 agonists with potential Th1-biasing activity. Analysis of a subset of these novel candidate Th1 type agonists in vivo in mice confirmed their ability to induce systemic cytokine responses consistent with a Th1 type bias. These results demonstrate the predictive value of this novel in vitro assay for assessing the in vivo functionality of glycolipid agonists and provide the basis for a relatively simple high-throughput assay for identification and functional classification of iNKT cell activating glycolipids. PMID:21425779

  2. A Rapid Fluorescence-Based Assay for Classification of iNKT Cell Activating Glycolipids

    PubMed Central

    2011-01-01

    Structural variants of ?-galactosylceramide (?GC) that activate invariant natural killer T cells (iNKT cells) are being developed as potential immunomodulatory agents for a variety of applications. Identification of specific forms of these glycolipids that bias responses to favor production of proinflammatory vs anti-inflammatory cytokines is central to current efforts, but this goal has been hampered by the lack of in vitro screening assays that reliably predict the in vivo biological activity of these compounds. Here we describe a fluorescence-based assay to identify functionally distinct ?GC analogues. Our assay is based on recent findings showing that presentation of glycolipid antigens by CD1d molecules localized to plasma membrane detergent-resistant microdomains (lipid rafts) is correlated with induction of interferon-? secretion and Th1-biased cytokine responses. Using an assay that measures lipid raft residency of CD1d molecules loaded with ?GC, we screened a library of ?200 synthetic ?GC analogues and identified 19 agonists with potential Th1-biasing activity. Analysis of a subset of these novel candidate Th1 type agonists in vivo in mice confirmed their ability to induce systemic cytokine responses consistent with a Th1 type bias. These results demonstrate the predictive value of this novel in vitro assay for assessing the in vivo functionality of glycolipid agonists and provide the basis for a relatively simple high-throughput assay for identification and functional classification of iNKT cell activating glycolipids. PMID:21425779

  3. Photobleaching-based quantitative analysis of fluorescence resonance energy transfer inside single living cell.

    PubMed

    Wang, Longxiang; Chen, Tongsheng; Qu, Junle; Wei, Xunbin

    2010-01-01

    The current advances of fluorescence microscopy and new fluorescent probes make fluorescence resonance energy transfer (FRET) a powerful technique for studying protein-protein interactions inside living cells. It is very hard to quantitatively analyze FRET efficiency using intensity-based FRET imaging microscopy due to the presence of autofluorescence and spectral crosstalks. In this study, we for the first time developed a novel photobleaching-based method to quantitatively detect FRET efficiency (Pb-FRET) by selectively photobleaching acceptor. The Pb-FRET method requires two fluorescence detection channels: a donor channel (CH ( 1 )) to selectively detect the fluorescence from donor, and a FRET channel (CH ( 2 )) which normally includes the fluorescence from both acceptor and donor due to emission spectral crosstalk. We used the Pb-FRET method to quantitatively measure the FRET efficiency of SCAT3, a caspase-3 indicator based on FRET, inside single living cells stably expressing SCAT3 during STS-induced apoptosis. At 0, 6 and 12 h after STS treatment, the FRET efficiency of SCAT3 obtained by Pb-FRET inside living cells was verified by two-photon excitation (TPE) fluorescence lifetime imaging microscopy (FLIM). The temporal resolution of Pb-FRET method is in second time-scale for ROI photobleaching, even in microsecond time-scale for spot photobleaching. Our results demonstrate that the Pb-FRET method is independent of photobleaching degree, and is very useful for quantitatively monitoring protein-protein interactions inside single living cell. PMID:19588234

  4. Highly sensitive fluorescent probe for clenbuterol hydrochloride detection based on its catalytic oxidation of eosine Y by NaIO4.

    PubMed

    Liu, Jiaming; Liu, Zhen-bo; Huang, Qitong; Lin, Chang-Qing; Lin, Xiaofeng

    2014-09-01

    A highly sensitive fluorescent probe for clenbuterol hydrochloride (CLB) detection has been first designed based on its catalytic effect on NaIO4 oxidating eosine Y (R). And this environment-friendly, simple, rapid, selective and sensitive fluorescent probe has been utilized to detect CLB in the practical samples with the results consisting with those obtained by GC/MS. The structures of R and CLB were characterized by infrared spectra. The mechanism of the proposed assay for the detection of CLB was also discussed. PMID:25155629

  5. A label-free fluorescent molecular beacon based on DNA-Ag nanoclusters for the construction of versatile Biosensors.

    PubMed

    Cao, Qiao; Teng, Ye; Yang, Xuan; Wang, Jin; Wang, Erkang

    2015-12-15

    In this paper, we developed a simple, low-cost and sensitive DNA sequences detection biosensor based on a label-free molecular beacon (MB) whose DNA hairpin structure terminal has a guanine-rich sequence that can enhance fluorescence of silver nanoclusters (Ag NCs). Without hybridization between hairpin probe and target DNA, the Ag NCs presented bright fluorescence for the proximity of guanine-rich sequences (GRSs). After binding with target DNA, the hairpin shape was destroyed which results in a decrease of the Ag NCs fluorescence intensity. With this biosensor, we detected three disease-related genes that were the human immunodeficiency virus (HIV) gene, hepatitis B virus (HBV) gene and human T-lymphotropic virus type I (HTLV-I) gene. The detection limits based on S/N of 3 were 4.4 nM, 6.8 nM and 8.5 nM for HIV gene, HBV gene and HTLV-I gene, respectively. Our sensor was also of high selectivity and could distinguish even one nucleotide mismatched target. PMID:26159151

  6. Solvent-assistant self-assembly of an AIE+TICT fluorescent Schiff base for the improved ammonia detection.

    PubMed

    Han, Tianyu; Wei, Wei; Yuan, Jing; Duan, Yuai; Li, Yaping; Hu, Liangyu; Dong, Yuping

    2016-04-01

    Solvent-assistant self-assembly of an AIE+TICT fluorescent Schiff base into one-dimensional nanofilaments has been developed. The orientation of the assemblies can be controlled by a simple dewetting process: the filaments are interweaved when the self-assembly process is performed on a horizontal substrate, while tilting the substrate to a tiny angle results in the formation of highly oriented ones with long-range order as verified by microscopic examination. The compound shows remarkable fluorescent response to ammonia gas based on a TICT-LE transition. The self-assembled film presents higher detection sensitivity compared with the non-assembled test paper: the former enables 4.75 times faster response time and 6.86 times lower detection limit than the latter. Furthermore, the former demonstrates better selectivity toward ammonia gas in the presence of various organic amines. The sensing devices also enjoy the advantage of cyclic utilization. The fluorescence of the fumed devices can be converted back into the original state when they are heated at 100°C for 5min, as thermal treatment can desorb the ammonia gas that adsorbed in the sensing devices. PMID:26838387

  7. Silver nanoclusters-based fluorescence assay of protein kinase activity and inhibition.

    PubMed

    Shen, Congcong; Xia, Xiaodong; Hu, Shengqiang; Yang, Minghui; Wang, Jianxiu

    2015-01-01

    A simple and sensitive fluorescence method for monitoring the activity and inhibition of protein kinase (PKA) has been developed using polycytosine oligonucleotide (dC12)-templated silver nanoclusters (Ag NCs). Adenosine-5'-triphosphate (ATP) was found to enhance the fluorescence of Ag NCs, while the hydrolysis of ATP to adenosine diphosphate (ADP) by PKA decreased the fluorescence of Ag NCs. Compared to the existing methods for kinase activity assay, the developed method does not involve phosphorylation of the substrate peptides, which significantly simplifies the detection procedures. The method exhibits high sensitivity, good selectivity, and wide linear range toward PKA detection. The inhibition effect of kinase inhibitor H-89 on the activity of PKA was also studied. The sensing protocol was also applied to the assay of drug-stimulated activation of PKA in HeLa cell lysates. PMID:25517425

  8. Reference Materials for Fluorescence Based on Inorganic Glass

    NASA Astrophysics Data System (ADS)

    Levin, A. D.; Pribytkov, V. A.; Nagaev, A. I.; Sadagov, A. Yu.

    Reference materials (RM) for relative spectral correction of emission spectra and day-to-day sensitivity monitoring of spectrofluorimeters were developed. The 2 kinds of inorganic glass were used as RM material - custom developed Cu+ -ion doped phosphate glass and colored optical glass SZS-17 (blue-green). RM can be either cuvette-shaped or in the form of flat plate and installed in sample compartment of the instrument. Flat plate geometry allows to minimize the dependency of RM fluorescence intensity from the characteristics of instrument's optical circuit due to inner filter effect.

  9. Highly sensitive naphthalimide-based fluorescence polarization probe for detecting cancer cells.

    PubMed

    Jia, Ti; Fu, Congying; Huang, Chusen; Yang, Haotian; Jia, Nengqin

    2015-05-13

    Fluorescence polarization (FP)-based signal is a self-referencing fluorescence signal, and it is less dependent on dye concentration and environmental interferences, which makes FP measurement an attractive alternative sensing technology to fluorescence intensity-based detection. However, most of the fluorescence polarization probes were constructed by introducing fluorescein, rhodamine, and cyanine dyes, which have relatively shorter excited-state lifetimes compared with BODIPY and naphthalimide dyes. Herein, a first naphthalimide based fluorescence polarization probe (BIO) was designed and synthesized for selective and direct detection of cancer cells. The relatively longer excited-state lifetimes and high photostability of naphthalimide makes BIO more sensitive and accuracy in quantitative determination of HeLa cells in homogeneous solution without cell lysis and further separation steps. The detection limit of BIO for HeLa cells was about 85 cells mL(-1), the linear range was from 2.5 10(2) cells mL(-1) to 1 10(6) cells mL(-1) and the response time is no more than 25 min. Moreover, due to the relatively high photostability of naphthalimide, BIO was particularly suitable for live cell imaging under continuous irradiation with confocal microscopy, and the specific interaction of BIO with CD44-overexpressing cell lines was clearly visualized. Importantly, this BIO based sensing platform offers a direct and real-time tool for cancer cell diagnosis when complemented with the use of naphthalimide-based fluorescence polarization probe. PMID:25898141

  10. Prospects for fluorescence based imaging/visualization of hydrodynamic systems on the National Ignition Facility

    SciTech Connect

    Suter, L. J., LLNL

    1998-06-04

    The next generation of large, high power lasers, such as the National Ignition Facility (NIF) [1] in the United States, Laser Mega Joule [2] in France or Helen Successor [3] in the United Kingdom offer the prospect of x-ray fluorescence based diagnosis of hydrodynamic experiments The x-ray fluorescence could be pumped by at least two techniques One technique is to use a sizable fraction of these facilities` high power to efficiently make multi-kilovolt x-rays which, in turn, causes dopants placed in experimental packages to fluoresce We call this ``externally pumped x-ray fluorescence`` The second technique is to use the sizable multi-kilovolt photon background that we expect to be present in many hohlraum based experiments, while the driving laser is on, to pump x-ray fluorescence The fluorescing medium could be a dopant in an experimental package or, possibly, a relatively thick slab of material in the hohlraum wall which could serve as a backlighter We call this ``hohlraum hot-corona pumped fluorescence``.

  11. Probes for biomolecules detection based on RET-enhanced fluorescence polarization.

    PubMed

    Ren, Dahai; Wang, Jun; Wang, Bin; You, Zheng

    2016-05-15

    Fluorescent probes based on the principle of resonance energy transfer (RET) or the principle of fluorescence polarization (FP) are already used to detect biomolecules independently. However, there were no in-depth studies about the impact of RET on FP. Also, very few studies gave a comprehensive analysis on how to effectively design such a fluorescent probe. Based on the principle of resonance energy transfer (RET), we constructed fluorescent probes (SA-488-sub-nanogold) using streptavidin labeled Alexa488 (SA-488), nanogold and biotinylated substrate peptide (biotin-subpeptide). The influence of the structure and the ingredients of the substrate peptide were discussed. After SA-488 was combined with the biotin-subpeptide and the nanogold, its fluorescence intensity (FI) would be suppressed due to the energy transfer, leading to an increase in its volume and mass. The suppression of the FI led to a decrease in SA-488's effective concentration, and the increase in the volume or mass prolonged the SA-488's rotational relaxation time. Both changes increased SA-488's polarization in the solution. Therefore, the FP performance of the probe is enhanced by the RET. Using the probe, trypsin and biotin were detected by the change in both fluorescence intensity and fluorescence polarization, showing higher reliability, higher sensitivity, and a lower detection limit. PMID:26774994

  12. Novel and remarkable enhanced-fluorescence system based on gold nanoclusters for detection of tetracycline.

    PubMed

    Yang, Xiaoming; Zhu, Shanshan; Dou, Yao; Zhuo, Yan; Luo, Yawen; Feng, Yuanjiao

    2014-05-01

    Tetracycline and Eu(3+), while coexisting, usually appear as a complex by chelating. This complex shows low fluorescence intensity, leading to its limitation of analytical goals. Gold nanoclusters (AuNCs), emerging as novel nano-material, are attracting increasing attentions in multiple fields. Herein, gold nanoclusters first function as a fluorescence-enhanced reagent rather than a conventional fluorescent-probe, and a dramatic enhanced-fluorescence system was built based on Eu(3+)-Tetracycline complex (EuTC) by introducing gold nanoclusters. Simultaneously, three types of gold nanoclusters were employed for exploring various conditions likely affecting the system, which demonstrate that no other gold nanoclusters than DNA-templated gold nanoclusters enormously caused fluorescence-enhancement of EuTC. Moreover, this enhanced-fluorescence system permitted available detection of tetracycline (TC) in a linear range of 0.01-5 ?M, with a detection limit of 4 nM at a signal-to-noise ratio of 3. Significantly, the practicality of this method for detection of TC in human urine and milk samples was validated, demonstrating its advantages of simplicity, sensitivity and low cost. Interestingly, this system described here is probably promising for kinds of applications based on its dramatically enhanced-fluorescence. PMID:24720959

  13. Label free selective detection of estriol using graphene oxide-based fluorescence sensor

    NASA Astrophysics Data System (ADS)

    Kushwaha, H. S.; Sao, Reshma; Vaish, Rahul

    2014-07-01

    Water-soluble and fluorescent Graphene oxide (GO) is biocompatible, easy, and economical to synthesize. Interestingly, GO is also capable of quenching fluorescence. On the basis of its fluorescence and quenching abilities, GO has been reported to serve as an energy acceptor in a fluorescence resonance energy transfer (FRET) sensor. GO-based FRET biosensors have been widely reported for sensing of proteins, nucleic acid, ATP (Adenosine triphosphate), etc. GO complexes with fluorescent dyes and enzymes have been used to sense metal ions. Graphene derivatives have been used for sensing endocrine-disrupting chemicals like bisphenols and chlorophenols with high sensitivity and good reproducibility. On this basis, a novel GO based fluorescent sensor has been successfully designed to detect estriol with remarkable selectivity and sensitivity. Estriol is one of the three estrogens in women and is considered to be medically important. Estriol content of maternal urine or plasma acts as an important screening marker for estimating foetal growth and development. In addition, estriol is also used as diagnostic marker for diseases like breast cancer, osteoporosis, neurodegenerative and cardiovascular diseases, insulin resistance, lupus erythematosus, endometriosis, etc. In this present study, we report for the first time a rapid, sensitive with detection limit of 1.3 nM, selective and highly biocompatible method for label free detection of estriol under physiological conditions using fluorescence assay.

  14. Boronate-Based Fluorescent Probes for Imaging Cellular Hydrogen Peroxide

    PubMed Central

    Miller, Evan W.; Albers, Aaron E.; Chang, Christopher J.; Pralle, Arnd; Isacoff, Ehud Y.

    2006-01-01

    The syntheses, properties, and biological applications of the Peroxysensor family, a new class of fluorescent probes for hydrogen peroxide, are presented. These reagents utilize a boronate deprotection mechanism to provide high selectivity and optical dynamic range for detecting H2O2 in aqueous solution over similar reactive oxygen species (ROS) including superoxide, nitric oxide, tert-butyl hydroperoxide, hypochlorite, singlet oxygen, ozone, and hydroxyl radical. Peroxyresorufin-1 (PR1), Peroxyfluor-1 (PF1), and Peroxyxanthone-1 (PX1) are first-generation probes that respond to H2O2 by an increase in red, green, and blue fluorescence, respectively. The boronate dyes are cell-permeable and can detect micromolar changes in H2O2 concentrations in living cells, including hippocampal neurons, using confocal microscopy and two-photon microscopy. The unique combination of ROS selectivity, membrane permeability, and a range of available excitation/emission colors establishes the potential value of PR1, PF1, PX1, and related probes for interrogating the physiology and pathology of cellular H2O2. PMID:16305254

  15. Molecular self assembly on optical fiber-based fluorescence sensor

    NASA Astrophysics Data System (ADS)

    Ayyagari, Madhu S. R.; Gao, Harry H.; Bihari, Bipin; Chittibabu, Kethinni G.; Kumar, Jayant; Marx, Kenneth A.; Kaplan, David L.; Tripathy, Sukant K.

    1994-03-01

    We discuss the molecular self-assembly on optical fibers in which a novel method for protein attachment to the sensing tip of the fiber is used. Our objective is to assemble a conjugated polythiophene copolymer as an attachment vehicle. Subsequent attachment of the photodynamic phycobiliprotein serves as the fluorescence probe element. Following our earlier experiments from Langmuir-Blodgett deposition of these polymeric materials as thin films on glass substrates, we extended the technique to optical fibers. First, the bare fiber surface is silanized with a C18 silane compound. The copolymer (3-undecylthiophene-co-3- methanolthiophene, biotinylated at the methanol moiety) assembly on the fiber is carried out presumable through van der Waals interactions between the hydrophobic fiber surface and the undecyl alkyl chains on the polymer backbone. A conjugated Str-PE (streptavidin covalently attached to phycoerythrin) complex is then attached to the copolymer via the conventional biotin-streptavidin interaction. The conjugated polymer not only supports the protein but, in principle, may help to transduce the signal generated by phycoerythrin to the fiber. Our results from fluorescence intensity measurements proved the efficacy of this system. An improved methodology is also sought to more strongly attach the conjugated copolymer to the fiber surface, and a covalent scheme is developed to polymerize and biotinylate polythiophene in situ on the fiber surface.

  16. A bio-aerosol detection technique based on tryptophan intrinsic fluorescence measurement

    NASA Astrophysics Data System (ADS)

    Cai, Shuyao; Zhang, Pei; Zhu, Linglin; Zhao, Yongkai; Huang, Huijie

    2011-12-01

    Based on the measurement of intrinsic fluorescence, a set of bio-aerosol including virus aerosols detection instrument is developed, with which a method of calibration is proposed using tryptophan as the target. The experimental results show a good linear relationship between the fluorescence voltage of the instrument and the concentration of the tryptophan aerosol. An excellent correlation (R2>=0.99) with the sensitivity of 4000PPL is obtained. The research demonstrates the reliability of the bio-aerosol detection by measuring the content of tryptophan. Further more the feasibility of prejudgment to the species of bio-aerosol particles with the multi-channel fluorescence detection technology is discussed.

  17. A rhodamine-based fluorescent probe for Cu(II) determination in aqueous solution.

    PubMed

    Mao, Jie; Cheng, Junhua; Wang, Xuchun; Wang, Shui; Cheng, Nianshou; Wang, Junfeng

    2015-03-01

    An 'off-on' rhodamine-based fluorescence probe for the selective detection of Cu(II) has been designed, exploiting the guest-induced structure transform mechanism. This system shows a sharp Cu(II)-selective fluorescence enhancement response in an aqueous system under physiological pH, and possesses high selectivity against a background of environmentally and biologically relevant metal ions. Under optimum conditions, the fluorescence intensity enhancement of this system is linearly proportional to the Cu(II) concentration from 50?nM to 6.0??M with a detection limit of 29 nM. PMID:25045042

  18. A dansyl-rhodamine ratiometric fluorescent probe for Hg2+ based on FRET mechanism.

    PubMed

    Xie, Puhui; Guo, Fengqi; Wang, Lingyu; Yang, Sen; Yao, Denghui; Yang, Guoyu

    2015-03-01

    Based on resonance energy transfer (FRET) from dansyl to rhodamine 101, a new fluorescent probe (compound 1) containing rhodamine 101 and a dansyl unit was synthesized for detecting Hg(2+) through ratiometric sensing in DMSO aqueous solutions. This probe shows a fast, reversible and selective response toward Hg(2+) in a wide pH range. Hg(2+) induced ring-opening reactions of the spirolactam rhodamine moiety of 1, leading to the formation of fluorescent derivatives that can serve as the FRET acceptors. Very large stokes shift (220nm) was observed in this case. About 97-fold increase in fluorescence intensity ratio was observed upon its binding with Hg(2+). PMID:25597044

  19. Micro-optical lens array for fluorescence detection in droplet-based microfluidics.

    PubMed

    Lim, Jiseok; Gruner, Philipp; Konrad, Manfred; Baret, Jean-Christophe

    2013-04-21

    We demonstrate the design and integration of droplet-based microfluidic devices with microoptical element arrays for enhanced detection of fluorescent signals. We show that the integration of microlenses and mirror surfaces in these devices results in an 8-fold increase in the fluorescence signal and in improved spatial resolution. Using an array of microlenses, massively parallel detection of droplets containing fluorescent dyes was achieved, leading to detection throughputs of about 2000 droplets per second and per lens, parallelized over 625 measurement points. PMID:23455606

  20. Phenylboronic acid functionalized reduced graphene oxide based fluorescence nano sensor for glucose sensing.

    PubMed

    Basiruddin, S K; Swain, Sarat K

    2016-01-01

    Reduced graphene has emerged as promising tools for detection based application of biomolecules as it has high surface area with strong fluorescence quenching property. We have used the concept of fluorescent quenching property of reduced graphene oxide to the fluorescent probes which are close vicinity of its surface. In present work, we have synthesized fluorescent based nano-sensor consist of phenylboronic acid functionalized reduced graphene oxide (rGO-PBA) and di-ol modified fluorescent probe for detection of biologically important glucose molecules. This fluorescent graphene based nano-probe has been characterized by high resolution transmission electron microscope (HRTEM), Atomic force microscope (AFM), UV-visible, Photo-luminescence (PL) and Fourier transformed infrared (FT-IR) spectroscopy. Finally, using this PBA functionalized reduced GO based nano-sensor, we were able to detect glucose molecule in the range of 2 mg/mL to 75 mg/mL in aqueous solution of pH7.4. PMID:26478292

  1. A fluorescence-based helicase assay: application to the screening of G-quadruplex ligands.

    PubMed

    Mendoza, Oscar; Gueddouda, Nassima Meriem; Boul, Jean-Baptiste; Bourdoncle, Anne; Mergny, Jean-Louis

    2015-06-23

    Helicases, enzymes that unwind DNA or RNA structure, are present in the cell nucleus and in the mitochondrion. Although the majority of the helicases unwind DNA or RNA duplexes, some of these proteins are known to resolve unusual structures such as G-quadruplexes (G4) in vitro. G4 may form stable barrier to the progression of molecular motors tracking on DNA. Monitoring G4 unwinding by these enzymes may reveal the mechanisms of the enzymes and provides information about the stability of these structures. In the experiments presented herein, we developed a reliable, inexpensive and rapid fluorescence-based technique to monitor the activity of G4 helicases in real time in a 96-well plate format. This system was used to screen a series of G4 structures and G4 binders for their effect on the Pif1 enzyme, a 5' to 3' DNA helicase. This simple assay should be adaptable to analysis of other helicases and G4 structures. PMID:25765657

  2. A fluorescence-based helicase assay: application to the screening of G-quadruplex ligands

    PubMed Central

    Mendoza, Oscar; Gueddouda, Nassima Meriem; Boul, Jean-Baptiste; Bourdoncle, Anne; Mergny, Jean-Louis

    2015-01-01

    Helicases, enzymes that unwind DNA or RNA structure, are present in the cell nucleus and in the mitochondrion. Although the majority of the helicases unwind DNA or RNA duplexes, some of these proteins are known to resolve unusual structures such as G-quadruplexes (G4) in vitro. G4 may form stable barrier to the progression of molecular motors tracking on DNA. Monitoring G4 unwinding by these enzymes may reveal the mechanisms of the enzymes and provides information about the stability of these structures. In the experiments presented herein, we developed a reliable, inexpensive and rapid fluorescence-based technique to monitor the activity of G4 helicases in real time in a 96-well plate format. This system was used to screen a series of G4 structures and G4 binders for their effect on the Pif1 enzyme, a 5? to 3? DNA helicase. This simple assay should be adaptable to analysis of other helicases and G4 structures. PMID:25765657

  3. Preparation, regulation and biological application of a Schiff base fluorescence probe.

    PubMed

    Yin, Ninghua; Diao, Haipeng; Liu, Wen; Wang, Jingru; Feng, Liheng

    2016-01-15

    A facile fluorescence switch with Schiff base units was designed and achieved by nucleophilic addition and dehydration reaction. The fluorescence of the probe can be regulated by metal ions (Al(3+) and Cu(2+)). The whole process shows that the weak fluorescence of the probe enhances with the addition of Al(3+), and then the strong fluorescence of the probe/Al(3+) ensemble reduces by introducing Cu(2+). Meanwhile, the solution color changes of the probe with metal ions can be observed under 365nm UV-vis light from weak light, pale green, green, pale green to weak light. Noticeably, the photo regulation processes of the probe by metal ions can be realized in the biological system and applied in cells imaging. The work provides a new strategy for designing facile regulation probe and develops a new application for Schiff base derivatives. PMID:26282317

  4. Fluorescent Ensemble Based on Bispyrene Fluorophore and Surfactant Assemblies: Sensing and Discriminating Proteins in Aqueous Solution.

    PubMed

    Fan, Junmei; Ding, Liping; Bo, Yu; Fang, Yu

    2015-10-14

    A particular bispyrene fluorophore (1) with two pyrene moieties covalently linked via a hydrophilic spacer was synthesized. Fluorescence measurements reveal that the fluorescence emission of 1 could be well modulated by a cationic surfactant, dodecyltrimethylammonium bromide (DTAB). Protein sensing studies illustrate that the selected ensemble based on 1/DTAB assemblies exhibits ratiometric responses to nonmetalloproteins and turn-off responses to metalloproteins, which can be used to differentiate the two types of proteins. Moreover, negatively charged nonmetalloproteins can be discriminated from the positively charged ones according to the difference in ratiometric responses. Fluorescence sensing studies with control bispyrenes indicate that the polarity of the spacer connecting two pyrene moieties plays an important role in locating bispyrene fluorophore in DTAB assemblies, which further influences its sensing behaviors to noncovalent interacting proteins. This study sheds light on the influence of the probe structure on the sensing performance of a fluorescent ensemble based on probe and surfactant assemblies. PMID:26414441

  5. Preparation, regulation and biological application of a Schiff base fluorescence probe

    NASA Astrophysics Data System (ADS)

    Yin, Ninghua; Diao, Haipeng; Liu, Wen; Wang, Jingru; Feng, Liheng

    2016-01-01

    A facile fluorescence switch with Schiff base units was designed and achieved by nucleophilic addition and dehydration reaction. The fluorescence of the probe can be regulated by metal ions (Al3 + and Cu2 +). The whole process shows that the weak fluorescence of the probe enhances with the addition of Al3 +, and then the strong fluorescence of the probe/Al3 + ensemble reduces by introducing Cu2 +. Meanwhile, the solution color changes of the probe with metal ions can be observed under 365 nm UV-vis light from weak light, pale green, green, pale green to weak light. Noticeably, the photo regulation processes of the probe by metal ions can be realized in the biological system and applied in cells imaging. The work provides a new strategy for designing facile regulation probe and develops a new application for Schiff base derivatives.

  6. A functionalized gold nanoparticles and Rhodamine 6G based fluorescent sensor for high sensitive and selective detection of mercury(II) in environmental water samples.

    PubMed

    Chen, Jinlong; Zheng, AiFang; Chen, AiHong; Gao, Yingchun; He, Chiyang; Kai, Xiaoming; Wu, Genhua; Chen, Youcun

    2007-09-01

    A gold-nanoparticles (Au NPs)-Rhodamine 6G (Rh6G) based fluorescent sensor for detecting Hg(II) in aqueous solution has been developed. Water-soluble and monodisperse gold nanoparticles (Au NPs) has been prepared facilely and further modified with thioglycolic acid (TGA). Free Rh6G dye was strongly fluorescent in bulk solution. The sensor system composing of Rh6G and Au NPs fluoresce weakly as result of fluorescence resonance energy transfer (FRET) and collision. The fluorescence of Rh6G and Au NPs based sensor was gradually recovered due to Rh6G units departed from the surface of functionalized Au NPs in the presence of Hg(II). Based on the modulation of fluorescence quenching efficiency of Rh6G-Au NPs by Hg(II) at pH 9.0 of teraborate buffer solution, a simple, rapid, reliable and specific turn-on fluorescent assay for Hg(II) was proposed. Under the optimum conditions, the fluorescence intensity of sensor is proportional to the concentration of Hg(II). The calibration graphs are linear over the range of 5.0x10(-10) to 3.55x10(-8) mol L(-1), and the corresponding limit of detection (LOD) is low as 6.0x10(-11) mol L(-1). The relative standard deviation of 10 replicate measurements is 1.5% for 2.0x10(-9) mol L(-1) Hg(II). In comparison with conventional fluorimetric methods for detection of mercury ion, the present nanosensor endowed with higher sensitivity and selectivity for Hg(II) in aqueous solution. Mercury(II) of real environmental water samples was determined by our proposed method with satisfactory results that were obtained by atomic absorption spectroscopy (AAS). PMID:17765073

  7. Selective fluorescence sensors for detection of nitroaniline and metal Ions based on ligand-based luminescent metal-organic frameworks

    NASA Astrophysics Data System (ADS)

    Yu, Zongchao; Wang, Fengqin; Lin, Xiangyi; Wang, Chengmiao; Fu, Yiyuan; Wang, Xiaojun; Zhao, Yongnan; Li, Guodong

    2015-12-01

    Metal-organic frameworks (MOFs) are porous crystalline materials with high potential for applications in fluorescence sensors. In this work, two solvent-induced Zn(II)-based metal-organic frameworks, Zn3L3(DMF)2 (1) and Zn3L3(DMA)2(H2O)3 (2) (L=4,4?-stilbenedicarboxylic acid), were investigated as selective sensing materials for detection of nitroaromatic compounds and metal ions. The sensing experiments show that 1 and 2 both exhibit selective fluorescence quenching toward nitroaniline with a low detection limit. In addition, 1 exhibits high selectivity for detection of Fe3+ and Al3+ by significant fluorescence quenching or enhancement effect. While for 2, it only exhibits significant fluorescence quenching effect for Fe3+. The results indicate that 1 and 2 are both promising fluorescence sensors for detecting and recognizing nitroaniline and metal ions with high sensitivity and selectivity.

  8. Microfluidics using a thiol-acrylate resin for fluorescence-based pathogen detection assays.

    PubMed

    Zhang, W; Tullier, M P; Patel, K; Carranza, A; Pojman, J A; Radadia, A D

    2015-11-01

    We demonstrate thiol-acrylate microfluidics prepared via soft lithography for single-step protein immobilization and fluorescence-based pathogen detection. Such microfluidics are formed via room temperature curing, and bonded without oxygen plasma. The background fluorescence of the resin was found to be similar to PDMS for several filter sets. We also show that thiol-acrylate devices are able to bond to gold-coated surfaces, which allows for integration with microfabricated sensors. PMID:26371689

  9. BODIPY based colorimetric fluorescent probe for selective thiophenol detection: theoretical and experimental studies.

    PubMed

    Kand, Dnyaneshwar; Mishra, Pratyush Kumar; Saha, Tanmoy; Lahiri, Mayurika; Talukdar, Pinaki

    2012-09-01

    A BODIPY-based selective thiophenol probe capable of discriminating aliphatic thiols is reported. The fluorescence off-on effect upon reaction with thiol is elucidated with theoretical calculations. The sensing of thiophenol is associated with a color change from red to yellow and 63-fold enhancement in green fluorescence. Application of the probe for selective thiophenol detection is demonstrated by live cell imaging. PMID:22751002

  10. An unnatural amino acid based fluorescent probe for phenylalanine ammonia lyase.

    PubMed

    Tian, Zhenlin; Zhu, Weiping; Xu, Yufang; Qian, Xuhong

    2014-08-21

    A fluorescent probe (2a-LP) based on an unnatural amino acid (UAA) is developed for the detection of phenylalanine ammonia lyase (PAL). In the presence of PAL, 2a-LP is catalytically deaminated to ortho-amino-transcinnamic acid (o-a-CA), which shows a remarkable offon fluorescence signal. Thus, the probe 2a-LP enables direct visualization of the PAL activity in tomato under UV illumination and has potential in vitro assays. PMID:24971756

  11. Room-temperature single-photon sources based on nanocrystal fluorescence in photonic/plasmonic nanostructures

    NASA Astrophysics Data System (ADS)

    Lukishova, S. G.; Winkler, J. M.; Bissell, L. J.; Mihaylova, D.; Liapis, Andreas C.; Shi, Z.; Goldberg, D.; Menon, V. M.; Boyd, R. W.; Chen, G.; Prasad, P.

    2014-10-01

    Results are presented here towards robust room-temperature SPSs based on fluorescence in nanocrystals: colloidal quantum dots, color-center diamonds and doped with trivalent rare-earth ions (TR3+). We used cholesteric chiral photonic bandgap and Bragg-reflector microcavities for single emitter fluorescence enhancement. We also developed plasmonic bowtie nanoantennas and 2D-Si-photonic bandgap microcavities. The paper also provides short outlines of other technologies for room-temperature single-photon sources.

  12. A flash-lamp based device for fluorescence detection and identification of individual pollen grains

    NASA Astrophysics Data System (ADS)

    Kiselev, Denis; Bonacina, Luigi; Wolf, Jean-Pierre

    2013-03-01

    We present a novel optical aerosol particle detector based on Xe flash lamp excitation and spectrally resolved fluorescence acquisition. We demonstrate its performances on three natural pollens acquiring in real-time scattering intensity at two wavelengths, sub-microsecond time-resolved scattering traces of the particles' passage in the focus, and UV-excited fluorescence spectra. We show that the device gives access to a rather specific detection of the bioaerosol particles.

  13. "Turn-off" fluorescent data array sensor based on double quantum dots coupled with chemometrics for highly sensitive and selective detection of multicomponent pesticides.

    PubMed

    Fan, Yao; Liu, Li; Sun, Donglei; Lan, Hanyue; Fu, Haiyan; Yang, Tianming; She, Yuanbin; Ni, Chuang

    2016-04-15

    As a popular detection model, the fluorescence "turn-off" sensor based on quantum dots (QDs) has already been successfully employed in the detections of many materials, especially in the researches on the interactions between pesticides. However, the previous studies are mainly focused on simple single track or the comparison based on similar concentration of drugs. In this work, a new detection method based on the fluorescence "turn-off" model with water-soluble ZnCdSe and CdSe QDs simultaneously as the fluorescent probes is established to detect various pesticides. The fluorescence of the two QDs can be quenched by different pesticides with varying degrees, which leads to the differences in positions and intensities of two peaks. By combining with chemometrics methods, all the pesticides can be qualitative and quantitative respectively even in real samples with the limit of detection was 2 × 10(-8) mol L(-1) and a recognition rate of 100%. This work is, to the best of our knowledge, the first report on the detection of pesticides based on the fluorescence quenching phenomenon of double quantum dots combined with chemometrics methods. What's more, the excellent selectivity of the system has been verified in different mediums such as mixed ion disruption, waste water, tea and water extraction liquid drugs. PMID:27016442

  14. A Model-based approach for microvasculature structure distortion correction in two-photon fluorescence microscopy images.

    PubMed

    Dao, Lam; Glancy, Brian; Lucotte, Bertrand; Chang, Lin-Ching; Balaban, Robert S; Hsu, Li-Yueh

    2015-11-01

    This paper investigates a postprocessing 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 modelling 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 subvolumes 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 (3D) 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

  15. A ratiometric fluorescent probe for aluminum ions based-on monomer/excimer conversion and its applications to real samples.

    PubMed

    Xie, Huiting; Wu, Yinglong; Huang, Jing; Zeng, Fang; Wu, Hao; Xia, Xitao; Yu, Changmin; Wu, Shuizhu

    2016-05-01

    Excessive amount of aluminum is detrimental to growing plants or animals and people are likely to suffer from various diseases upon long-term exposure to aluminum ions. Therefore, sensitive and selective detection of trace amounts of Al(3+) in real samples is of great importance. Herein, a ratiometric fluorescent probe for detecting aluminum ions based on pyrene-1-butyric acid (Py-L-COOH) was developed, which function via monomer/excimer conversion. In the presence of Al(3+), the original monomer-state Py-L-COOH molecules coordinate with Al(3+) and form excimer, thus changing the emission wavelength from 350-400nm to 450-500nm and achieving the ratiometric detection for Al(3+). The probe responds to Al(3+) quickly and can be operable in aqueous media with a very low detection limit of 0.29µM. This system is capable of detecting Al(3+) in real samples and shows high selectivity. Furthermore, the probe exhibits low cytotoxicity and can be used in fluorescence imaging in Hela cell lines. The approach may provide an effective simple probe for aluminum ions determination with application to real samples, as well as offering insights for designing facile ratiometric fluorescent sensors. PMID:26946004

  16. Fluorescence detection in capillary arrays based on galvanometer step scanning.

    PubMed

    Xue, G; Yeung, E S

    2001-10-01

    A computer-controlled galvanometer scanner is adapted for scanning a focused laser beam across a 96-capillary array for laser-induced fluorescence detection. The signal at a single photomultiplier tube is temporally sorted to distinguish among the capillaries. The limit of detection for fluoresceins is 3 x 10(-11) M (S/N = 3) for 5 mW of total laser power scanned at 4 Hz. The observed cross-talk among capillaries is 0.2%. Advantages include the efficient utilization of light due to the high duty-cycle of step scan, good detection performance due to the reduction of stray light, ruggedness due to the small mass of the galvanometer mirror, low cost due to the simplicity of components, and flexibility due to the independent paths for excitation and emission. PMID:11669531

  17. Fluorescent sensor based on a novel conjugated polyfluorene derivative.

    PubMed

    Gao, Weiqiang; Yan, Mei; Ge, Shenguang; Liu, Xiaoxia; Yu, Jinghua

    2012-09-01

    A novel water-soluble polyfluorene derivative, poly[(9,9-bis(3'-((N,N-dimethylamino)N-ethylammonium)propyl)-2,7-fluorene)-alt-2,7-(9,9-p-divinylbenzene)]dibromide (P-2) was synthesized by the palladium-catalyzed Suzuki coupling reaction and it's quaternized ammonium polyelectrolyte derivatives was obtained through a postpolymerization treatment on the terminal amino groups. The electrochemical and optical properties of the copolymers was fully investigated. The results showed that the new polyfluorene derivative had high electronic conductivity and strong fluorescence, therefore it had good potential to be used in chemical and biological sensors, as shown in optical sensing of bovine albumin (BSA) in this study. PMID:22634413

  18. Fluorescent sensor based on a novel conjugated polyfluorene derivative

    NASA Astrophysics Data System (ADS)

    Gao, Weiqiang; Yan, Mei; Ge, Shenguang; Liu, Xiaoxia; Yu, Jinghua

    2012-09-01

    A novel water-soluble polyfluorene derivative, poly[(9,9-bis(3'-((N,N-dimethylamino)N-ethylammonium)propyl)-2,7-fluorene)-alt-2,7-(9,9-p-divinylbenzene)]dibromide (P-2) was synthesized by the palladium-catalyzed Suzuki coupling reaction and it's quaternized ammonium polyelectrolyte derivatives was obtained through a postpolymerization treatment on the terminal amino groups. The electrochemical and optical properties of the copolymers was fully investigated. The results showed that the new polyfluorene derivative had high electronic conductivity and strong fluorescence, therefore it had good potential to be used in chemical and biological sensors, as shown in optical sensing of bovine albumin (BSA) in this study.

  19. Fluorescent-based chemical sensor for organophosphate detection

    NASA Astrophysics Data System (ADS)

    Urek, pela Korent; Lobnik, Aleksandra

    2011-05-01

    We present a new optical sensor for the detection of organophosphates by incorporating fluorescent indicator dye into sol-gel material. We used different configurations of immobilization matrices such as thin film and spherical nanoparticles. The sensor thin films were prepared by using acid-catalyzed sol-gel process and the spherical nanoparticles by modified Stber method. The effects of configuration matrices on the sensor's characteristic were studied. The use of dye-doped nanoparticles improved the detection limit from 0.69 ?M to 17 nM, response time from 600 s to 12 s, precision and sensitivity, but reduced the sensor's working rage from 6.910-7 M - 6.910-3 M to 1.7510-8M - 2.310-7 M.

  20. Effects of alcohols on fluorescence intensity and color of a discharged-obelin-based biomarker.

    PubMed

    Alieva, Roza R; Belogurova, Nadezhda V; Petrova, Alena S; Kudryasheva, Nadezhda S

    2014-05-01

    Photoproteins are responsible for bioluminescence of marine coelenterates; bioluminescent and fluorescent biomarkers based on photoproteins are useful for monitoring of calcium-dependent processes in medical investigations. Here, we present the analysis of intensity and color of light-induced fluorescence of Ca(2+)-discharged photoprotein obelin in the presence of alcohols (ethanol and glycerol). Complex obelin spectra obtained at different concentrations of the alcohols at 350- and 280-nm excitation (corresponding to polypeptide-bound coelenteramide and tryptophan absorption regions) were deconvoluted into Gaussian components; fluorescent intensity and contributions of the components to experimental spectra were analyzed. Five Gaussian components were found in different spectral regions-ultraviolet (tryptophan emission), blue-green (coelenteramide emission), and red (hypothetical indole-coelenteramide exciplex emission). Inhibition coefficients and contributions of the components to experimental fluorescent spectra showed that presence of alcohols increased contributions of ultraviolet, violet, and red components, but decreased contributions of components in the blue-green region. The effects were related to (1) changes of proton transfer efficiency in fluorescent S*1 state of coelenteramide in the obelin active center and (2) formation of indole-coelenteramide exciplex at 280-nm photoexcitation. The data show that variation of fluorescence color and intensity in the presence of alcohols and dependence of emission spectra on excitation wavelength should be considered while applying the discharged obelin as a fluorescence biomarker. PMID:24618986

  1. Synthesis and Fluorescence Properties of Pyrimidine-Based Diboron Complexes with Donor-?-Acceptor Structures.

    PubMed

    Kubota, Yasuhiro; Kasatani, Kouhei; Niwa, Takahiro; Sato, Hiroyasu; Funabiki, Kazumasa; Matsui, Masaki

    2016-01-01

    Pyrimidine-based diboron complexes bearing ?-iminoenolate ligands and phenyl groups as bulky substituents on the boron atoms were synthesized as novel fluorescent dyes, and their fluorescence properties were investigated in solution and in the solid state. The diboron complexes with donor-?-acceptor structures showed positive solvatochromism in the fluorescence spectra. The cyano derivative exhibited the most dramatic redshift of the fluorescence maximum Fmax with increasing solvent polarity (from 551?nm in n-hexane to 710?nm in acetonitrile). The diboron complexes showed solid-state fluorescence in the range of 578-706?nm with fluorescence quantum yields of 0.06-0.28. Additionally, the trifluoromethyl derivative exhibited solvent-inclusion solid-state fluorescence. The trifluoromethyl derivative formed toluene-inclusion and ethyl acetate-inclusion crystals. The toluene-inclusion crystal (Fmax =668?nm, ?f =0.16) showed a blueshifted Fmax and higher ?f value compared to the original trifluoromethyl derivative (Fmax =694?nm, ?f =0.08) in the solid state. On the other hand, the Fmax (709?nm) and ?f (0.04) values of the ethyl acetate-inclusion crystal were redshifted and lower, respectively. PMID:26670268

  2. A fluorescence-based hydrolytic enzyme activity assay for quantifying toxic effects of Roundup to Daphnia magna.

    PubMed

    rsted, Michael; Roslev, Peter

    2015-08-01

    Daphnia magna is a widely used model organism for aquatic toxicity testing. In the present study, the authors investigated the hydrolytic enzyme activity of D. magna after exposure to toxicant stress. In vivo enzyme activity was quantified using 15 fluorogenic enzyme probes based on 4-methylumbelliferyl or 7-amino-4-methylcoumarin. Probing D. magna enzyme activity was evaluated using short-term exposure (24-48 h) to the reference chemical K2 Cr2 O7 or the herbicide formulation Roundup. Toxicant-induced changes in hydrolytic enzyme activity were compared with changes in mobility (International Organization for Standardization standard 6341). The results showed that hydrolytic enzyme activity was quantifiable as a combination of whole body fluorescence of D. magna and the fluorescence of the surrounding water. Exposure of D. magna to lethal and sublethal concentrations of Roundup resulted in loss of whole body enzyme activity and release of cell constituents, including enzymes and DNA. Roundup caused comparable inhibition of mobility and alkaline phosphatase activity with median effective concentration values at 20?C of 8.7?mg active ingredient (a.i.)/L to 11.7?mg?a.i./L. Inhibition of alkaline phosphatase activity by Roundup was lowest at 14?C and greater at 20?C and 26?C. The results suggest that the fluorescence-based hydrolytic enzyme activity assay (FLEA assay) can be used as an index of D. magna stress. Combining enzyme activity with fluorescence measurements may be applied as a simple and quantitative supplement for toxicity testing with D. magna. PMID:25809520

  3. Diketopyrrolopyrrole: brilliant red pigment dye-based fluorescent probes and their applications.

    PubMed

    Kaur, Matinder; Choi, Dong Hoon

    2015-01-01

    The development of fluorescent probes for the detection of biologically relevant species is a burgeoning topic in the field of supramolecular chemistry. A number of available dyes such as rhodamine, coumarin, fluorescein, and cyanine have been employed in the design and synthesis of new fluorescent probes. However, diketopyrrolopyrrole (DPP) and its derivatives have a distinguished role in supramolecular chemistry for the design of fluorescent dyes. DPP dyes offer distinctive advantages relative to other organic dyes, including high fluorescence quantum yields and good light and thermal stability. Significant advancements have been made in the development of new fluorescent probes based on DPP in recent years as a result of tireless research efforts by the chemistry scientific community. In this tutorial review, we highlight the recent progress in the development of DPP-based fluorescent probes for the period spanning 2009 to the present time and the applications of these probes to recognition of biologically relevant species including anions, cations, reactive oxygen species, thiols, gases and other miscellaneous applications. This review is targeted toward providing the readers with deeper understanding for the future design of DPP-based fluorogenic probes for chemical and biological applications. PMID:25186723

  4. Comparison of fluorescence-based semi-automated genotyping of multiple microsatellite loci with autoradiographic techniques

    SciTech Connect

    Schwengel, D.A.; Jedlicka, A.E.; Levitt, R.C.

    1994-07-01

    The practical application of highly efficient fluorescence-based methods for the semi-automated genotyping of polymerase chain reaction-based microsatellite markers will depend on the development of robust protocols that provide accurate and reproducible data. In the present report the authors compare the accuracy of a fluorescence-based protocol with a benchmark radiolabeling method that depends on a known sequence ladder or amplified DNA from reference individuals for sizing by autoradiography. Three microsatellite markers, IGF (mfd1), D4S174 (mfd 59), and D5S211 (mfd 154), with products overlapping in size were each labeled with a different fluorophore and run simultaneously with an internal size standard in a single electrophoretic lane. The size of each allele was compared for these markers by using both techniques for five larger CEPH families (884, 1331, 1333, and 1362). Of 462 possible alleles, four discrepancies (0.8%) were identified when the two approaches were compared. The authors conclude that the fluorescence-based protocol is at least as accurate as the standard radiolabeling technique since none of the sizing errors arose as a result of the fluorescence-based technique. They describe the adaptation of the fluorescence-based protocol to the simultaneous analysis of up to 24 microsatellite loci per electrophoretic lane. These highly accurate and efficient semi-automated techniques will be useful in high-resolution genomic analyses. 18 refs., 4 tabs., 3 figs.

  5. Assessment of dental demineralization of yellow race based on fluorescence spectrum

    NASA Astrophysics Data System (ADS)

    Zhan, Zhenlin; Chen, Chuanguo; Li, Xuwei; Zhang, Xianzeng; Xie, Shusen

    2014-11-01

    The goal of this study was to evaluate the demineralization status at different acid-etch time based on fluorescence spectrum. Human molars in vitro of yellow race were cut into tooth sections and then they were immersed in 0.3% citric acid to simulate the oral natural demineralization. According to the acid-etch time, samples were randomly divided into three groups: I:20 min, II:40 min, and III:60 min. The normal untreated specimen was set as control group. The fluorescence spectra before and after treatment were measured and analyzed. The result showed that fluorescence spectrum could be efficiently used to monitor the demineralization status of human dental tissue. The relative fluorescence intensities of dental tissue excited respectively with 260, 330 and 400 nm decreased with the increase of acid-etch time, though there was no new constituent formed after demineralization.

  6. Bioimaging of nitric oxide with fluorescent indicators based on the rhodamine chromophore.

    PubMed

    Kojima, H; Hirotani, M; Nakatsubo, N; Kikuchi, K; Urano, Y; Higuchi, T; Hirata, Y; Nagano, T

    2001-05-01

    Diaminofluoresceins are widely used for detection and imaging of nitric oxide (NO), but for biological applications, they have the disadvantages that the fluorescence of the fluorescein chromophore is pH-sensitive and overlaps the autofluorescence of cells. We have developed a membrane-permeable fluorescent indicator for NO based on the rhodamine chromophore, DAR-4M AM, which can be excited with 550-nm light. The fluorescence quantum yield of the product after reaction with NO is 840 times higher than that of DAR-4M. The detection limit of NO was 7 nM, and the fluorescence showed no pH dependency above pH 4. DAR-4M AM was successfully applied to practical bioimaging of NO produced in bovine aortic endothelial cells. PMID:11354477

  7. A rhodamine based fluorescent probe for Hg2+ and its application to cellular imaging.

    PubMed

    Yan, Fanyong; Cao, Donglei; Yang, Ning; Wang, Meng; Dai, Linfeng; Li, Chuying; Chen, Li

    2013-04-01

    A new rhodamine-based fluorescent probe (Rh-F) for detection of Hg(2+) ions was synthesized, which could bind Hg(2+) in aqueous ethanol (7:3, v/v) at pH 7.0 with detectable change in color and fluorescence. The response is based on a ring opening reaction and formation of a 1:1 complex, while ring-opening process of spirolactam enables large fluorescent enhancement and colorimetric change upon the addition of Hg(2+). The response is reversible and linear in the range between 200nM and 1000nM, with a detection limit of 4.2nM. Selectivity and competition experiments with various other metal ion revealed that Rh-F possesses highly selective fluorescent response to Hg(2+). Furthermore, the probe was successfully applied to fluorescent imaging of Hg(2+) in L-929 cells confirm that Rh-F can be used as a fluorescent probe for monitoring Hg(2+) in living cells. PMID:23353763

  8. Long wavelength fluorescence based biosensors for in vivo continuous monitoring of metabolites

    NASA Astrophysics Data System (ADS)

    Thomas, Joseph; Ambroise, Arounaguiry; Birchfield, Kara; Cai, Wensheng; Sandmann, Christian; Singh, Sarabjit; Weidemaier, Kristin; Pitner, J. Bruce

    2006-02-01

    The early stage development studies of novel implantable continuous metabolite sensor systems for glucose, lactate and fatty acids are discussed. These sensors utilize non-enzymatic "reagentless" sensor systems based on NIR fluorophore-labeled binding proteins. For in vivo applications, NIR fluorescence based systems (beyond 600 nm) have the added benefit of reduced interference from background scattering, tissue and serum absorption and cell auto-fluorescence. The long wavelength emission facilitates implanted sensor disks to transmit fluorescence to an external reader through wireless connections and the resulting fluorescence signals can be correlated to metabolite concentrations. We have developed a prototype optical system that uses a bifurcated optical fiber to transmit excitation and read emission at the surface of the skin. With this system, fluorescence signals were read over time through animal skin. The changes in glucose concentration were studied using immobilized sensor proteins and were compared to non-immobilized sensors in solution. For sensors in solution, no response delay was observed. For immobilized systems, the fluorescence response showed a delay corresponding to the diffusion time for the metabolite to equilibrate within the sensor.

  9. Characterization of photophysical and base-mimicking properties of a novel fluorescent adenine analogue in DNA

    PubMed Central

    Dierckx, Anke; Dinr, Peter; El-Sagheer, Afaf H.; Kumar, Joshi Dhruval; Brown, Tom; Grtli, Morten; Wilhelmsson, L. Marcus

    2011-01-01

    To increase the diversity of fluorescent base analogues with improved properties, we here present the straightforward click-chemistry-based synthesis of a novel fluorescent adenine-analogue triazole adenine (AT) and its photophysical characterization inside DNA. AT shows promising properties compared to the widely used adenine analogue 2-aminopurine. Quantum yields reach >20% and >5% in single- and double-stranded DNA, respectively, and show dependence on neighbouring bases. Moreover, AT shows only a minor destabilization of DNA duplexes, comparable to 2-aminopurine, and circular dichroism investigations suggest that AT only causes minimal structural perturbations to normal B-DNA. Furthermore, we find that AT shows favourable base-pairing properties with thymine and more surprisingly also with normal adenine. In conclusion, AT shows strong potential as a new fluorescent adenine analogue for monitoring changes within its microenvironment in DNA. PMID:21278417

  10. Seminaphthofluorescein-Based Fluorescent Probes for Imaging Nitric Oxide in Live Cells

    PubMed Central

    Pluth, Michael D.; Chan, Maria R.; McQuade, Lindsey E.; Lippard, Stephen J.

    2011-01-01

    Fluorescent turn-on probes for nitric oxide based on seminaphthofluorescein scaffolds were prepared and spectroscopically characterized. The Cu(II) complexes of these fluorescent probes react with NO under anaerobic conditions to yield a 20- to 45-fold increase in integrated emission. The seminaphthofluorescein-based probes emit at longer wavelengths than the parent FL1 and FL2 fluorescein-based generations of NO probes, maintaining emission maxima between 550 and 625 nm. The emission profiles depend on the excitation wavelength; maximum fluorescence turn-on is achieved at excitations between 535575 nm. The probes are highly selective for NO over other biologically relevant reactive nitrogen and oxygen species including NO3 ?, NO2 ?, HNO, ONOO?, NO2, OCl?, and H2O2. The seminaphthofluorescein-based probes can be used to visualize endogenously produced NO in live cells as demonstrated using Raw 264.7 macrophages. PMID:21895023

  11. Fluorescence anisotropy-based structure-switching aptamer assay using a peptide nucleic acid (PNA) probe.

    PubMed

    Goux, Emma; Lespinasse, Quentin; Guieu, Valérie; Perrier, Sandrine; Ravelet, Corinne; Fiore, Emmanuelle; Peyrin, Eric

    2016-03-15

    This study describes for the first time the feasibility of using peptide nucleic acids (PNAs) as an alternative to the DNA probes in structure-switching aptamer fluorescence polarisation assays. The effects of experimental parameters such as the length of the PNA strand, the nature of dye and the buffer conditions on the assay performances are first explored using two different methodologies based on the competition between the PNA/aptamer hydribridisation and the target/aptamer complexation. d-ATP can be detected from 1 to 25μM in a linear range and a detection limit (LOD) of 3μM can be reached. For this target, this lowers by a factor >5 the LOD reported with conventional DNA-based fluorescent structure switching aptamer-based assays and by a factor 3 the LOD observed with non-competitive fluorescent sensing platform indicating the usefulness of the PNA-based approach. PMID:26455538

  12. When Simple Harmonic Motion Is Not that Simple: Managing Epistemological Complexity by Using Computer-Based Representations

    ERIC Educational Resources Information Center

    Parnafes, Orit

    2010-01-01

    Many real-world phenomena, even "simple" physical phenomena such as natural harmonic motion, are complex in the sense that they require coordinating multiple subtle foci of attention to get the required information when experiencing them. Moreover, for students to develop sound understanding of a concept or a phenomenon, they need to learn to get

  13. A simple method for the multi-elemental analysis of organic fertilizer by slurry sampling and total reflection X-ray fluorescence.

    PubMed

    Resende, Luciene V; Nascentes, Clsia C

    2016-01-15

    A simple and fast method for the multi-elemental determination of 18 inorganic constituents (P, S, Cl, K, Ca, Ti, Cr, V, Mn, Fe, Ni, Cu, Zn, Br, Rb, Sr, Ba and Pb) in organic fertilizers employing slurry sampling and total reflection X-ray fluorescence (TXRF) is presented. A 2(3) factorial design with a central point was employed to optimize the slurry sampling procedure. The internal standard and instrumental conditions were optimized by univariate studies. The selectivity of the method to determining Se, As, Pb, Cr, Ni and Cd was assessed. The accuracy was evaluated by the analysis of four standard reference materials (SRM). The recoveries varied from 72% to 114%. For most of the elements, good agreement was achieved between the certified value and the value measured in the SRM. The relative standard deviation (RSD %) ranged from 0.5% to 14%. The evaluated method was applied to the determination of analytes in the press cake of palm, castor, curcas, sunflower, fodder turnip, white lupin, rapeseed and pequi, and their potential to be used as organic fertilizer was evaluated in accordance with Brazilian legislation. PMID:26592637

  14. Simple method for the simultaneous isolation and determination of fumonisin B1 and its metabolite aminopentol-1 in swine liver by liquid chromatography--fluorescence detection.

    PubMed

    Pagliuca, Giampiero; Zironi, Elisa; Ceccolini, Alberto; Matera, Riccardo; Serrazanetti, Gian Paolo; Piva, Andrea

    2005-05-01

    An analytical method based on high-performance liquid chromatography (HPLC) combined with fluorescence detection (FL) has been developed for the simultaneous determination of fumonisin B1 (FB1) and its totally hydrolized metabolite aminopentol-1 (AP1) in pig liver. The sample preparation is based on a single solid phase extraction (SPE). o-Phthalaldehyde (OPA) was used for pre-column derivatization before the programmed reversed-phase analysis on phenylhexyl column. The developed method shows good repeatibility for inter- and intra-day precision as well as adequate linearity of calibration curves (r2 was 0.9855 for FB1 and 0.9831 for AP1). Average recoveries from the matrix were 93.6% for FB1 and 95.3% for AP1. The limit of quantification (LOQ) in swine liver was 75 microg/kg for FB1 and 42 microg/kg for AP1. PMID:15797527

  15. Cyanine-based probe\\tag-peptide pair for fluorescence protein imaging and fluorescence protein imaging methods

    DOEpatents

    Mayer-Cumblidge, M. Uljana; Cao, Haishi

    2010-08-17

    A molecular probe comprises two arsenic atoms and at least one cyanine based moiety. A method of producing a molecular probe includes providing a molecule having a first formula, treating the molecule with HgOAc, and subsequently transmetallizing with AsCl.sub.3. The As is liganded to ethanedithiol to produce a probe having a second formula. A method of labeling a peptide includes providing a peptide comprising a tag sequence and contacting the peptide with a biarsenical molecular probe. A complex is formed comprising the tag sequence and the molecular probe. A method of studying a peptide includes providing a mixture containing a peptide comprising a peptide tag sequence, adding a biarsenical probe to the mixture, and monitoring the fluorescence of the mixture.

  16. A fast reconstruction method for fluorescence molecular tomography based on improved iterated shrinkage

    NASA Astrophysics Data System (ADS)

    Han, Dong; Tian, Jie; Qin, Chenghu; Zhang, Bo; Liu, Kai; Ma, Xibo

    2011-03-01

    Fluorescence molecular tomography (FMT) has become a promising imaging modality for in vivo small animal molecular imaging, and has many successful applications. This is partly due to the wealth of the fluorescent probes. By labeling the regions of interest with fluorescent probes, FMT can achieve non-invasive investigation of the biological process by localizing the targeted probes based on certain inverse mathematical models. However, FMT is usually an illposed problem, and some form of regularization should be included to stabilize the problem, which can be considered as the a priori information of the fluorescent probe bio-distribution. When FMT is used for the early detection of tumors, an important characteristic is the sparsity of the fluorescent sources. This is because tumors are usually very small and sparse at this stage. Considering this, general sparsity-promoting Lp-norm regularization is utilized in this paper. The iterated shrinkage based reconstruction method is adopted to solve the general Lp regularization problem. However, the original iterated shrinkage method is proved to have a linear convergence rate, and a large number of iterations are needed to obtain satisfactory results. In this paper, an improved iterated shrinkage based FMT reconstruction algorithm is proposed. By using the solutions from two previous iterations to determine the current solution, the convergence rate can be greatly increased. Heterogeneous simulation experiment shows that the proposed method can obtain comparable results with greatly reduced number of iterations compared with the original iterated shrinkage based method, which makes it a practical reconstruction algorithm.

  17. Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform

    NASA Astrophysics Data System (ADS)

    Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

    2015-11-01

    Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr05839b

  18. Data of a fluorescent imaging-based analysis of anti-cancer drug effects on three-dimensional cultures of breast cancer cells

    PubMed Central

    Itou, Junji; Tanaka, Sunao; Li, Wenzhao; Matsumoto, Yoshiaki; Sato, Fumiaki; Toi, Masakazu

    2015-01-01

    Three-dimensional (3D) cell culture is a powerful tool to study cell growth under 3D condition. To perform a simple test for anti-cancer drugs in 3D culture, visualization of non-proliferated cells is required. We propose a fluorescent imaging-based assay to analyze cancer cell proliferation in 3D culture. We used a pulse-labeling technique with a photoconvertible fluorescent protein Kaede to identify non-proliferated cells. This assay allows us to observe change in cell proliferation in 3D culture by simple imaging. Using this assay, we obtained the data of the effects of anti-cancer drugs, 5-fluorouracil and PD0332991 in a breast cancer cell line, MCF-7.

  19. 10 CFR 429.35 - Bare or covered (no reflector) medium base compact fluorescent lamps.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 3 2014-01-01 2014-01-01 false Bare or covered (no reflector) medium base compact....35 Bare or covered (no reflector) medium base compact fluorescent lamps. (a) Sampling plan for selection of units for testing. (1) The requirements of 429.11 are applicable to bare or covered...

  20. 10 CFR 429.35 - Bare or covered (no reflector) medium base compact fluorescent lamps.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 3 2012-01-01 2012-01-01 false Bare or covered (no reflector) medium base compact....35 Bare or covered (no reflector) medium base compact fluorescent lamps. (a) Sampling plan for selection of units for testing. (1) The requirements of 429.11 are applicable to bare or covered...

  1. 10 CFR 429.35 - Bare or covered (no reflector) medium base compact fluorescent lamps.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 3 2013-01-01 2013-01-01 false Bare or covered (no reflector) medium base compact....35 Bare or covered (no reflector) medium base compact fluorescent lamps. (a) Sampling plan for selection of units for testing. (1) The requirements of 429.11 are applicable to bare or covered...

  2. Development of a Fluorescence Polarization-Based Diagnostic Assay for Equine Infectious Anemia Virus

    PubMed Central

    Tencza, Sarah Burroughs; Islam, Kazi R.; Kalia, Vandana; Nasir, Mohammad S.; Jolley, Michael E.; Montelaro, Ronald C.

    2000-01-01

    The control of equine infectious anemia virus (EIAV) infections of horses has been over the past 20 years based primarily on the identification and elimination of seropositive horses, predominantly by a standardized agar gel immunodiffusion (AGID) assay in centralized reference laboratories. This screening for EIAV-seropositive horses has been to date hindered by the lack of a rapid diagnostic format that can be easily employed in the field. We describe here the development of a rapid solution-phase assay for the presence of serum antibodies to EIAV based on fluorescence polarization (FP) (patent pending). Peptides derived from antigenic regions of EIAV core and envelope proteins were initially screened for their utility as probes in an FP assay to select the best peptide antigen candidates. The FP assay was optimized to detect the presence of EIAV-specific antibodies by a change in the FP of a fluorescein-labeled immunoreactive peptide diagnostic antigen. The most sensitive and specific peptide probe was a peptide corresponding to the immunodominant region of the EIAV transmembrane protein, gp45. This probe was tested for its reactivity in the optimized FP assay with 151 AGID-positive horse sera and 106 AGID-negative serum samples. The results of these studies demonstrated that the FP assay reactivity correlated with reported AGID results in 106 of 106 negative serum samples (100% specificity) and in 135 of 151 positive serum samples (89.4% sensitivity). The FP assay was also found to have a very low background reactivity and to readily detect antibodies produced early in infection (?3 weeks postinfection). The developed EIAV FP assay is rapid (5 to 20 min) and simple to perform and is equally suitable for use both in the field and in the diagnostic laboratory, thus providing the basis of an improved commercial diagnostic assay for EIAV infection of horses. PMID:10790112

  3. Multicolor Fluorescence Writing Based on Host-Guest Interactions and Force-Induced Fluorescence-Color Memory.

    PubMed

    Matsunaga, Yuki; Yang, Jye-Shane

    2015-06-26

    A new strategy is reported for multicolor fluorescence writing on thin solid films with mechanical forces. This concept is illustrated by the use of a green-fluorescent pentiptycene derivative 1, which forms variably colored fluorescent exciplexes: a change from yellow to red was observed with anilines, and fluorescence quenching (a change to black) occurred in the presence of benzoquinone. Mechanical forces, such as grinding and shearing, induced a crystalline-to-amorphous phase transition in both the pristine and guest-adsorbed solids that led to a change in the fluorescence color (mechanofluorochromism) and a memory of the resulting color. Fluorescence drawings of five or more colors were created on glass or paper and could be readily erased by exposure to air and dichloromethane fumes. The structural and mechanistic aspects of the observations are also discussed. PMID:25982228

  4. [Lake algae chemotaxonomy technology based on fluorescence excitation emission matrix and parallel factor analysis].

    PubMed

    Chen, Xiao-Na; Han, Xiu-Rong; Su, Rong-Guo; Shi, Xiao-Yong

    2014-03-01

    An in vivo three-dimensional fluorescence method for the determination of algae community structure was developed by parallel factor (PARAFAC) analysis and CHEMTAX. The PARAFAC model was applied to fluorescence excitation-emission matrix (EEM) of 23 algae species and 12 fluorescent components were identified according to the residual sum of squares and specificity of the composition profiles of fluorescent. Based on the 12 fluorescent components, the algae species at different growth stages were correctly classified at the division level using Bayesian discriminant analysis (BDA). Then the reference fluorescent component ratio matrix was constructed for CHEMTAX, and the EEM-PARAFAC-CHEMTAX method was developed to differentiate taxonomic groups of algae. When the fluorometric method was used for 531 single-species samples, the average correct discrimination ratio (CDR) was 99.1% and the correct discrimination ratios (CDRs) were 100% at the division level except Chlorophyta, the CDR of which was 97.5%. The CDRs for 95 mixtures were above 98.5% for the dominant algae species and above 90.5% for the subdominant algae species, with average relative contents of 69.7% and 26.4%, respectively. This technique would be of great aid when low-cost and rapid analysis is needed for samples in a large batch. PMID:24881379

  5. Highly sensitive and selective fluorescent assay for guanine based on the Cu(2+)/eosin Y system.

    PubMed

    Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling

    2016-05-15

    A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu(2+)/eosin Y. Cu(2+) interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu(2+)/eosin Y system, guanine reacted with Cu(2+) to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5nmolL(-1) and a linear range of 3.3-116nmolL(-1). The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe. PMID:26971024

  6. Use of Time-Resolved Fluorescence to Monitor Bioactive Compounds in Plant Based Foodstuffs.

    PubMed

    Lemos, M Adlia; Srnikov, Katarna; Bot, Francesca; Anese, Monica; Hungerford, Graham

    2015-01-01

    The study of compounds that exhibit antioxidant activity has recently received much interest in the food industry because of their potential health benefits. Most of these compounds are plant based, such as polyphenolics and carotenoids, and there is a need to monitor them from the field through processing and into the body. Ideally, a monitoring technique should be non-invasive with the potential for remote capabilities. The application of the phenomenon of fluorescence has proved to be well suited, as many plant associated compounds exhibit fluorescence. The photophysical behaviour of fluorescent molecules is also highly dependent on their microenvironment, making them suitable probes to monitor changes in pH, viscosity and polarity, for example. Time-resolved fluorescence techniques have recently come to the fore, as they offer the ability to obtain more information, coupled with the fact that the fluorescence lifetime is an absolute measure, while steady state just provides relative and average information. In this work, we will present illustrative time-resolved measurements, rather than a comprehensive review, to show the potential of time-resolved fluorescence applied to the study of bioactive substances. The aim is to help assess if any changes occur in their form, going from extraction via storage and cooking to the interaction with serum albumin, a principal blood transport protein. PMID:26132136

  7. Wideband fluorescence-based thermometry by neural network recognition: Photothermal application with 10 ns time resolution

    NASA Astrophysics Data System (ADS)

    Liu, Liwang; Zhong, Kuo; Munro, Troy; Alvarado, Salvador; Cte, Renaud; Creten, Sebastiaan; Fron, Eduard; Ban, Heng; Van der Auweraer, Mark; Roozen, N. B.; Matsuda, Osamu; Glorieux, Christ

    2015-11-01

    Neural network recognition of features of the fluorescence spectrum of a thermosensitive probe is exploited in order to achieve fluorescence-based thermometry with an accuracy of 200 mK with 100 MHz bandwidth, and with high robustness against fluctuations of the probe laser intensity used. The concept is implemented on a rhodamine B dyed mixture of copper chloride and glycerol, and the temperature dependent fluorescence is investigated in the temperature range between 234 K and 311 K. The spatial dependence of the calibrated amplitude and phase of photothermally induced temperature oscillations along the axis of the excitation laser are determined at different modulation frequencies. The spatial and frequency dependence of the extracted temperature signals is well fitted by a 1D multi-layer thermal diffusion model. In a time domain implementation of the approach, the gradual temperature rise due to the accumulation of the DC component of the heat flux supplied by repetitive laser pulses as well the immediate transient temperature evolution after each single pulse is extracted from acquired temporal sequences of fluorescence spectra induced by a CW green laser. A stroboscopic implementation of fluorescence thermometry, using a pulsed fluorescence evoking probe laser, is shown to achieve remote detection of temperature changes with a time resolution of 10 ns.

  8. Modulation of a fluorescence switch based on photochromic spirooxazine in composite organic nanoparticles

    NASA Astrophysics Data System (ADS)

    Sheng, Xiaohai; Peng, Aidong; Fu, Hongbing; Liu, Yuanyuan; Zhao, Yongsheng; Ma, Ying; Yao, Jiannian

    2007-04-01

    We describe a versatile and convenient approach to achieve fluorescence modulation by the preparation of composite nanoparticles (CNPs), based on photochromic 5-methoxy-1,3,3-trimethyl-9'-hydroxyspiroindolinenaphthoxazine (SO), fluorescent 4-(dicyanomethylene)-2-methyl-6-(p-dimethyl-aminostyryl)-4H-pyran (DCM), and emissive-assistant 1,3-bis(pyrene) propane (BPP) molecules, employing doping techniques. The mechanism of the fluorescence switch is the intermolecular energy transfer as supported by both steady-state and time-resolved spectroscopy results. The addition of BPP not only enhances the contrast of the fluorescence signal between the 'ON' and 'OFF' state, but also provides a convenient way to tune the excitation wavelength for reading the fluorescence. High-contrast ON/OFF (20:1) fluorescence switching is successfully implemented in the CNPs and also in a more practical PVA film loaded with the CNPs. This system may represent an alternative to the covalent system in potentially rewritable high-density optical data or image storage utilizing luminescence intensity readout schemes.

  9. FMN-Based Fluorescent Proteins as Heavy Metal Sensors Against Mercury Ions.

    PubMed

    Ravikumar, Yuvaraj; Nadarajan, Saravanan Prabhu; Lee, Chong-Soon; Jung, Seunho; Bae, Dong-Ho; Yun, Hyungdon

    2016-03-28

    Bacterial light-oxygen-voltage-sensing photoreceptor-derived flavin mononucleotide (FMN)- based fluorescent proteins act as a promising distinct class of fluorescent proteins utilized for various biomedical and biotechnological applications. The key property of its independency towards oxygen for its chromophore maturation has greatly helped this protein to outperform the other fluorescent proteins such as GFP and DsRed for anaerobic applications. Here, we describe the feasibility of FMN-containing fluorescent protein FbFP as a metal-sensing probe by measuring the fluorescence emission changes of a protein with respect to the concentration of metal ions. In the present study, we demonstrated the mercury-sensing ability of FbFP protein and the possible amino acids responsible for metal binding. A ratiometric approach was employed here in order to exploit the fluorescence changes observed at two different emission maxima with respect to Hg(2+) at micromolar concentration. The engineered variant FbFPC56I showed high sensitivity towards Hg(2+) and followed a good linear relationship from 0.1 to 3 µM of Hg(2+). Thus, further engineering with a rational approach would enable the FbFP to be developed as a novel and highly selective and sensitive biosensor for other toxic heavy metal ions as well. PMID:26699753

  10. Use of Time-Resolved Fluorescence to Monitor Bioactive Compounds in Plant Based Foodstuffs

    PubMed Central

    Lemos, M. Adília; Sárniková, Katarína; Bot, Francesca; Anese, Monica; Hungerford, Graham

    2015-01-01

    The study of compounds that exhibit antioxidant activity has recently received much interest in the food industry because of their potential health benefits. Most of these compounds are plant based, such as polyphenolics and carotenoids, and there is a need to monitor them from the field through processing and into the body. Ideally, a monitoring technique should be non-invasive with the potential for remote capabilities. The application of the phenomenon of fluorescence has proved to be well suited, as many plant associated compounds exhibit fluorescence. The photophysical behaviour of fluorescent molecules is also highly dependent on their microenvironment, making them suitable probes to monitor changes in pH, viscosity and polarity, for example. Time-resolved fluorescence techniques have recently come to the fore, as they offer the ability to obtain more information, coupled with the fact that the fluorescence lifetime is an absolute measure, while steady state just provides relative and average information. In this work, we will present illustrative time-resolved measurements, rather than a comprehensive review, to show the potential of time-resolved fluorescence applied to the study of bioactive substances. The aim is to help assess if any changes occur in their form, going from extraction via storage and cooking to the interaction with serum albumin, a principal blood transport protein. PMID:26132136

  11. A PDMS-Based Cylindrical Hybrid Lens for Enhanced Fluorescence Detection in Microfluidic Systems

    PubMed Central

    Lin, Bor-Shyh; Yang, Yu-Ching; Ho, Chong-Yi; Yang, Han-Yu; Wang, Hsiang-Yu

    2014-01-01

    Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS) to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 ?g/mL and 0.05 ?g/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light. PMID:24531300

  12. Two fluorescent Schiff base sensors for Zn(2+): the Zn(2+)/Cu(2+) ion interference.

    PubMed

    Jiménez-Sánchez, Arturo; Ortíz, Benjamín; Ortiz Navarrete, Vianney; Farfán, Norberto; Santillan, Rosa

    2015-09-01

    Two simple and low cost 2,4-di-tert-butyl-6-[(1-hydroxycyclohexylmethylimino)methyl]phenol (L1) and 2-[{(1-hydroxycyclohexyl)methylimino}methyl]phenol (L2) Schiff base sensors exhibiting selectivity for Zn(2+) in water:methanol (95:5, v/v, 10 mM HEPES) are described. L1 and L2 display an "off-on" fluorescence effect forming the L1·Zn and L2·Zn complexes, respectively. In the case of L1·Zn, the emission response is quenched by the addition of Cu(2+) forming the respective L1·Cu complex; in spite of that, the fluorescence signal can be completely restored only by the addition of tartrate anions (C4H4O6(2-)) forming again L1·Znvia the "off-on" displacement approach. However, in the case of L2·Zn no Cu(2+) interference is observed, which is a typical problem for Zn(2+) sensors. Here we describe that a very subtle structural change in the ligand during transition from the enol-imine tautomer in L1 to the keto-enamine tautomer in L2 is enough to modulate the Zn(2+)/Cu(2+) selectivity. Also, the Zn(2+)vs. Cd(2+) discrimination for L1 and L2 is proved. Moreover, we found that the interaction between both L·Zn complexes and tartrate anions completely restored the free ligands by the ligand substitution mechanism even in a more efficient association than phosphate anions. Further, a second colorimetric response channel upon addition of Fe(2+) was observed for L1 and L2. Then, TD-DFT theoretical calculations were conducted in order to study the efficiency of the sensors to give different responses in the presence of such metal ions. Finally, the L2 sensor successfully detects Zn(2+) in Jurkat cells cultured with and without Zn(2+) enriched medium. PMID:26192046

  13. Sandwich Immunoassays of Multicomponent Subtrace Pathogenic DNA Based on Magnetic Fluorescent Encoded Nanoparticles

    PubMed Central

    Liu, Yaxu; Zhang, Xuanjun; E, Yifeng; Fang, Fang; Kuang, Guangkai; Wang, Guannan

    2016-01-01

    A novel magnetic fluorescent encoded nanoimmunoassay system for multicomponent detection and separation of the subtrace pathogenic DNA (hepatitis B virus surface gene, HBV; hepatitis A virus poly the protein gene, HAV) was established based on new type of magnetic fluorescent encoded nanoparticles and sandwich immunoassay principle. This method combines multifunctional nanoparticles, immunoassay technique, fluorescence labeling, and magnetic separation of multicomponent technology. It has many advantages such as high sensitivity, low detection limit, easy operation, and great potential for development. The results of this work show that, based on nanoimmunoassay system, it could quantitatively detect the multicomponent trace pathogenic HAV and HBV DNA, as well as detection limit up to 0.1 pM and 0.12 pM. Furthermore, with the improvement of the performances of magnetic fluorescent encoded nanoparticles, the sensitivity will be further improved. In this experiment, a new nanoimmunoassay system based on magnetic fluorescent encoded nanoparticles was established, which will provide a new way for the immunoassay and separation of multicomponent biomolecules. PMID:26881227

  14. Fluorescent directed heteroduplex analysis enhances PCR-based DQA1 and DQB1 genotyping

    SciTech Connect

    Zimmerman, P.A.; Mansfield, E.S.; Miyasaki, T.

    1994-09-01

    We previously showed how directed heteroduplex analysis (DHDA) simplifies DQA1 and DQB1 genotyping and have used the technique to identify a new DQA1 allele (DQA{sup *}0502, which has a single nucleotide difference from DQA1{sup *}0501). In DHDA, labeled probes are mixed with unlabeled PCR products amplified from patient genomic DNA. After controlled re-annealing, allelic heteroduplexes are resolved on polyacrylamide gels (5%, 2.7 M urea). To utilize fluorescence imaging for detecting the heteroduplexes in HLA-typing, probes are labeled by PCR amplification using locus-specific generic primers and gels scanned using the Fluorimager{trademark} 575 (Molecular Dynamics, Inc.). We generate 2-color DHDA probes using locus-specific PCR primers 5{prime}-end labeled with the fluorochromes FAM (positive-strand primer) and JOE (negative-strand primer) (Perkin-Elmer). Genotypic analysis within families obtained from the CEPH repository have been performed by fluorescence-based DHDA. Results to date show 100% concordance between DHDA and sequence-specific oligonucleotide probe (SSOP) genotyping. Fluorescence-based DHDA is performed with fewer probes than SSOP (1 set of locus-specific probes for DHDA and 10 SSOP probes for DQA1 typing or 13 SSOP probes for DQB1 typing). In addition, fluorescent DHDA allows rapid assessment of genotype, aproximately four hours from receipt of sample to typing result. These results suggest that fluorescent DHDA may facilitate DNA-based HLA-typing within the time constraints required for solid organ transplantation.

  15. A rapid technique for classifying phytoplankton fluorescence spectra based on self-organizing maps.

    PubMed

    Aymerich, Ismael F; Piera, Jaume; Soria-Frisch, Aureli; Cros, Lluïsa

    2009-06-01

    Fluorescence spectroscopy has been demonstrated to be a powerful tool for characterizing phytoplankton communities in marine environments. Using different fluorescence spectra techniques, it is now possible to discriminate the major phytoplankton groups. However, most of the current techniques are based on fluorescence excitation measurements, which require stimulation at different wavelengths and thus considerable time to obtain the complete spectral profile. This requirement may be an important constraint for several mobile oceanographic platforms, such as vertical profilers or autonomous underwater vehicles, which require rapid-acquisition instruments. This paper presents a novel technique for classifying fluorescence spectra based on self-organizing maps (SOMs), one of the most popular artificial neural network (ANN) methods. The method is able to achieve phytoplankton discrimination using only fluorescence emission spectra (single wavelength excitation), thus reducing the acquisition time. The discrimination capabilities of SOM using excitation and emission spectra are compared. The analysis shows that the SOM has a good performance using excitation spectra, whereas data preprocessing is required in order to obtain similar discrimination capabilities using emission spectra. The final results obtained using emission spectra indicate that the discrimination is properly achieved even between algal groups, such as diatoms and dinoflagellates, which cannot be discriminated with previous methods. We finally point out that although techniques based on excitation spectra can achieve a better taxonomic accuracy, there are some applications that require faster acquisition processes. Acquiring emission spectra is almost instantaneous, and techniques such as SOM can achieve good classification performance using appropriately preprocessed data. PMID:19531300

  16. Sandwich Immunoassays of Multicomponent Subtrace Pathogenic DNA Based on Magnetic Fluorescent Encoded Nanoparticles.

    PubMed

    Liu, Yaxu; Zhang, Xuanjun; E, Yifeng; Fang, Fang; Kuang, Guangkai; Wang, Guannan

    2016-01-01

    A novel magnetic fluorescent encoded nanoimmunoassay system for multicomponent detection and separation of the subtrace pathogenic DNA (hepatitis B virus surface gene, HBV; hepatitis A virus poly the protein gene, HAV) was established based on new type of magnetic fluorescent encoded nanoparticles and sandwich immunoassay principle. This method combines multifunctional nanoparticles, immunoassay technique, fluorescence labeling, and magnetic separation of multicomponent technology. It has many advantages such as high sensitivity, low detection limit, easy operation, and great potential for development. The results of this work show that, based on nanoimmunoassay system, it could quantitatively detect the multicomponent trace pathogenic HAV and HBV DNA, as well as detection limit up to 0.1?pM and 0.12?pM. Furthermore, with the improvement of the performances of magnetic fluorescent encoded nanoparticles, the sensitivity will be further improved. In this experiment, a new nanoimmunoassay system based on magnetic fluorescent encoded nanoparticles was established, which will provide a new way for the immunoassay and separation of multicomponent biomolecules. PMID:26881227

  17. Fluorescence-Based Bioassays for the Detection and Evaluation of Food Materials

    PubMed Central

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-01-01

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR)-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA)-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials. PMID:26473869

  18. Fluorescence-based bioassays for the detection and evaluation of food materials.

    PubMed

    Nishi, Kentaro; Isobe, Shin-Ichiro; Zhu, Yun; Kiyama, Ryoiti

    2015-01-01

    We summarize here the recent progress in fluorescence-based bioassays for the detection and evaluation of food materials by focusing on fluorescent dyes used in bioassays and applications of these assays for food safety, quality and efficacy. Fluorescent dyes have been used in various bioassays, such as biosensing, cell assay, energy transfer-based assay, probing, protein/immunological assay and microarray/biochip assay. Among the arrays used in microarray/biochip assay, fluorescence-based microarrays/biochips, such as antibody/protein microarrays, bead/suspension arrays, capillary/sensor arrays, DNA microarrays/polymerase chain reaction (PCR)-based arrays, glycan/lectin arrays, immunoassay/enzyme-linked immunosorbent assay (ELISA)-based arrays, microfluidic chips and tissue arrays, have been developed and used for the assessment of allergy/poisoning/toxicity, contamination and efficacy/mechanism, and quality control/safety. DNA microarray assays have been used widely for food safety and quality as well as searches for active components. DNA microarray-based gene expression profiling may be useful for such purposes due to its advantages in the evaluation of pathway-based intracellular signaling in response to food materials. PMID:26473869

  19. Portable in situ fluorescence cytometry of microscale cell-based assays

    NASA Astrophysics Data System (ADS)

    Tatosian, Daniel A.; Shuler, Michael L.; Kim, Donghyun

    2005-07-01

    A portable fluorescence cytometric system has been developed for characterizing chemical concentration and cellular status in microscale cell culture analog (°CCA) devices. Based on discrete optical components, the system provides a modular platform for real-time image measurements applicable to a variety of cell-based microassays. As a feasibility study, we investigated the real-time dynamics of daunorubicin uptake with cultured mouse L-cells in a °CCA compartment. Time course results measured by the portable fluorescence cytometric system confirmed that in the °CCA devices daunorubicin accumulation is proportional to the liquid turnover rate.

  20. Fluorescence and excited state dynamics of the deprotonated Schiff base retinal in proteorhodopsin.

    PubMed

    Bhl, Elena; Braun, Markus; Lakatos, Andrea; Glaubitz, Clemens; Wachtveitl, Josef

    2015-09-01

    The UV light absorbing species of proteorhodopsin with deprotonated Schiff base retinal was investigated using steady-state fluorescence and femtosecond pump-probe spectroscopy. Compared to the all-trans retinal with protonated Schiff base, the deprotonated chromophore absorbs at 365 nm and exhibits a blue-shifted fluorescence spectrum. The unusually long-lived excited state decays bi-exponentially with time constants of 8 ps and 130 ps to form a deprotonated 13-cis retinal as the primary photo-product. PMID:26083266

  1. A highly sensitive and class-specific fluorescence polarisation assay for sulphonamides based on dihydropteroate synthase.

    PubMed

    Wang, Zhanhui; Liang, Xiao; Wen, Kai; Zhang, Suxia; Li, Chenglong; Shen, Jianzhong

    2015-08-15

    We describe a fluorescence polarisation assay based on the use of dihydropteroate synthase (DHPS) and a fluorescence probe for multi-sulphonamide detection. Dihydropteridine pyrophosphate (DHPPP) was synthesised and acts as the first substrate for DHPS. Under optimised conditions, the half-maximal inhibitory concentrations (IC50) of the assay were less than 100 ng mL(-1) for at least 29 sulphonamides and the time needed for the detection was less than 20 min. More importantly, the assay revealed quite uniform affinities for all of the individual sulphonamides tested, which has never before been achieved in an antibody-based assay. PMID:25775967

  2. Conventional and photonic crystal fiber based two-photon fluorescence biosensing

    NASA Astrophysics Data System (ADS)

    Myaing, Mon Thiri

    Optical fiber probes are widely used in the biomedical field for applications such as optical microscopy, endoscopy, and optical biopsy. Due to their flexibility and small size, optical fibers offer a minimally invasive light interface for imaging and spectroscopic analysis of internal tissue. The development of fluorescent probes for studies of biological processes has increased the importance of developing optical methods for quantitative, in vivo diagnosis. In this dissertation, we discuss the development of a novel two-photon optical fiber fluorescence (TPOFF) probe for real time, in vivo, quantitative fluorescence measurements in biological samples. In order to understand and optimize two-photon excitation through an optical fiber, pulse propagation effects must be considered. We found a simple phenomenological scaling behavior for the energy dependence of the pulse width for negatively pre-chirped pulses propagating in a normally dispersive fiber. As a consequence of this scaling behavior, the dependence of two-photon fluorescence (TPF) on the pulse intensity becomes sub-quadratic. The TPOFF probe employs a scheme where the same single-mode fiber (SMF) is used for both the excitation and collection of TPF. Using this fiber probe, we show quantification of tumor fluorescence both ex vivo and in vivo. In ex vivo measurements of tumors developed from cells expressing the green fluorescence protein (GFP), the TPOFF probe detected fluorescence from tumors with as little as 0.3% GFP cells. These results were similar to flow cytometry analysis of isolated cells from the tumors. The TPOFF measurements of GFP tumors in live, anesthetized mice showed a linear relationship between the measured fluorescence and the percentage of GFP expressing cells. The TPOFF probe was also used in targeted binding experiments of Herceptin antibody and folic acid-dendrimer nanoparticle conjugates. To improve the sensitivity of the TPOFF probe, a double-clad photonic crystal fiber (DCF) was employed. This fiber combines the advantages of both single mode fibers (high excitation efficiencies) and multimode fibers (high collection efficiencies). When we compare the through-fiber TPF signal from a Rhodamine dye gel collected by an SMF and DCF, we observe over an order of magnitude signal enhancement.

  3. Facile and Sensitive Fluorescence Sensing of Alkaline Phosphatase Activity with Photoluminescent Carbon Dots Based on Inner Filter Effect.

    PubMed

    Li, Guoliang; Fu, Huili; Chen, Xuejie; Gong, Peiwei; Chen, Guang; Xia, Lian; Wang, Hua; You, Jinmao; Wu, Yongning

    2016-03-01

    A simple and sensitive fluorescent assay for detecting alkaline phosphatase (ALP) based on the inner filter effect (IFE) has been proven, which is conceptually different from the previously reported ALP fluorescent assays. In this sensing platform, N-doped carbon dots (CDs) with a high quantum yield of 49% were prepared by one-pot synthesis and were directly used as a fluorophore in IFE. p-Nitrophenylphosphate (PNPP) was employed to act as an ALP substrate, and its enzyme catalytic product (p-nitrophenol (PNP)) was capable of functioning as a powerful absorber in IFE to influence the excitation of fluorophore (CDs). When in the presence of ALP, PNPP was transformed into PNP and induced the absorption band transition from 310 to 405 nm, which resulted in the complementary overlap between the absorption of PNP and the excitation of CDs. Because of the competitive absorption, the excitation of CDs was significantly weakened, resulting in the quenching of CDs. The present IFE-based sensing strategy showed a good linear relationship from 0.01 to 25 U/L (R(2) = 0.996) and provided an exciting detection limit of 0.001 U/L (signal-to-noise ratio of 3). The proposed sensing approach was successfully applied to ALP sensing in serum samples, ALP inhibitor investigation and phosphatase cell imaging. The presented IFE-based CDs fluorescence sensing strategy gives new insight on the development of the facile and sensitive optical probe for enzyme activity assay because the surface modification or the linking between the receptor and the fluorophore is no longer required. PMID:26820049

  4. A Simple System for Observing Dynamic Phase Equilibrium via an Inquiry-Based Laboratory or Demonstration

    ERIC Educational Resources Information Center

    Cloonan, Carrie A.; Andrew, Julie A.; Nichol, Carolyn A.; Hutchinson, John S.

    2011-01-01

    This article describes an activity that can be used as an inquiry-based laboratory or demonstration for either high school or undergraduate chemistry students to provide a basis for understanding both vapor pressure and the concept of dynamic phase equilibrium. The activity includes a simple setup to create a closed system of only water liquid and

  5. Two Simple Classroom Demonstrations for Scanning Probe Microscopy Based on a Macroscopic Analogy

    ERIC Educational Resources Information Center

    Hajkova, Zdenka; Fejfar, Antonin; Smejkal, Petr

    2013-01-01

    This article describes two simple classroom demonstrations that illustrate the principles of scanning probe microscopy (SPM) based on a macroscopic analogy. The analogy features the bumps in an egg carton to represent the atoms on a chemical surface and a probe that can be represented by a dwarf statue (illustrating an origin of the prefix…

  6. Two Simple Classroom Demonstrations for Scanning Probe Microscopy Based on a Macroscopic Analogy

    ERIC Educational Resources Information Center

    Hajkova, Zdenka; Fejfar, Antonin; Smejkal, Petr

    2013-01-01

    This article describes two simple classroom demonstrations that illustrate the principles of scanning probe microscopy (SPM) based on a macroscopic analogy. The analogy features the bumps in an egg carton to represent the atoms on a chemical surface and a probe that can be represented by a dwarf statue (illustrating an origin of the prefix

  7. A Simple System for Observing Dynamic Phase Equilibrium via an Inquiry-Based Laboratory or Demonstration

    ERIC Educational Resources Information Center

    Cloonan, Carrie A.; Andrew, Julie A.; Nichol, Carolyn A.; Hutchinson, John S.

    2011-01-01

    This article describes an activity that can be used as an inquiry-based laboratory or demonstration for either high school or undergraduate chemistry students to provide a basis for understanding both vapor pressure and the concept of dynamic phase equilibrium. The activity includes a simple setup to create a closed system of only water liquid and…

  8. A simple fluorometric assay for DNA exonuclease activity based on graphene oxide.

    PubMed

    Lee, Jieon; Min, Dal-Hee

    2012-05-01

    A new assay platform for DNA exonuclease activity is developed based on the preferential binding of single-stranded DNA (ssDNA) over double-stranded DNA (dsDNA) to graphene oxide. This approach allows a simple and quantitative activity measurement in a short time at low cost. PMID:22413127

  9. Evaluation of a Fluorescence-Based Method for Antibabesial Drug Screening

    PubMed Central

    Guswanto, Azirwan; Sivakumar, Thillaiampalam; Rizk, Mohamed Abdo; Elsayed, Shimaa Abd Elsalam; Youssef, Mohamed Ahmed; ElSaid, ElSaid El Shirbini; Yokoyama, Naoaki

    2014-01-01

    In vitro evaluation of chemotherapeutic agents against Babesia and Theileria parasites has become routine, and the effectiveness of these chemicals is usually determined by comparing the parasitemia dynamics of untreated and treated parasites. Although microscopy is widely used to calculate parasitemia, several disadvantages are associated with this technique. The present study evaluated a fluorescence-based method using SYBR green I stain (SG I) to screen antibabesial agents in in vitro cultures of Babesia bovis. The linearity between relative fluorescence units (RFU) and parasitemia was found to be well correlated with a 0.9944 goodness-of-fit (r2) value. Subsequently, 50% inhibitory concentration (IC50) values were calculated for 3 antiprotozoan agents, diminazene aceturate, nimbolide, and gedunin, by this method. For diminazene aceturate and nimbolide, the IC50s determined by the fluorescence-based method (408 nM and 8.13 ?M, respectively) and microscopy (400.3 nM and 9.4 ?M, respectively) were in agreement. Furthermore, the IC50 of gedunin determined by the fluorescence-based method (19 ?M) was similar to the recently described microscopy-based value (21.7 ?M) for B. bovis. Additionally, the Z? factor (0.80 to 0.90), signal-to-noise (S/N) ratio (44.15 to 87.64), coefficient of variation at the maximum signal (%CVmax) (0.50 to 2.85), and coefficient of variation at the minimum signal (%CVmin) (1.23 to 2.21) calculated for the fluorescence method using diminazene aceturate were comparable to those previously determined in malaria research for this assay. These findings suggest that the fluorescence-based method might be useful for antibabesial drug screening and may have potential to be developed into a high-throughput screening (HTS) assay. PMID:24914124

  10. Evaluation of a fluorescence-based method for antibabesial drug screening.

    PubMed

    Guswanto, Azirwan; Sivakumar, Thillaiampalam; Rizk, Mohamed Abdo; Elsayed, Shimaa Abd Elsalam; Youssef, Mohamed Ahmed; ElSaid, ElSaid El Shirbini; Yokoyama, Naoaki; Igarashi, Ikuo

    2014-08-01

    In vitro evaluation of chemotherapeutic agents against Babesia and Theileria parasites has become routine, and the effectiveness of these chemicals is usually determined by comparing the parasitemia dynamics of untreated and treated parasites. Although microscopy is widely used to calculate parasitemia, several disadvantages are associated with this technique. The present study evaluated a fluorescence-based method using SYBR green I stain (SG I) to screen antibabesial agents in in vitro cultures of Babesia bovis. The linearity between relative fluorescence units (RFU) and parasitemia was found to be well correlated with a 0.9944 goodness-of-fit (r(2)) value. Subsequently, 50% inhibitory concentration (IC50) values were calculated for 3 antiprotozoan agents, diminazene aceturate, nimbolide, and gedunin, by this method. For diminazene aceturate and nimbolide, the IC(50)s determined by the fluorescence-based method (408 nM and 8.13 ?M, respectively) and microscopy (400.3 nM and 9.4 ?M, respectively) were in agreement. Furthermore, the IC50 of gedunin determined by the fluorescence-based method (19 ?M) was similar to the recently described microscopy-based value (21.7 ?M) for B. bovis. Additionally, the Z' factor (0.80 to 0.90), signal-to-noise (S/N) ratio (44.15 to 87.64), coefficient of variation at the maximum signal (%CVmax) (0.50 to 2.85), and coefficient of variation at the minimum signal (%CVmin) (1.23 to 2.21) calculated for the fluorescence method using diminazene aceturate were comparable to those previously determined in malaria research for this assay. These findings suggest that the fluorescence-based method might be useful for antibabesial drug screening and may have potential to be developed into a high-throughput screening (HTS) assay. PMID:24914124

  11. High-efficiency white organic light-emitting diodes based on a blue thermally activated delayed fluorescent emitter combined with green and red fluorescent emitters.

    PubMed

    Higuchi, Takahiro; Nakanotani, Hajime; Adachi, Chihaya

    2015-03-25

    A new device architecture for highly efficient white organic light-emitting diodes is proposed, using a molecule exhibiting blue thermally activated delayed fluorescence as a common source of singlet excitons for molecules emitting red and green light based on conventional fluorescence. The device, with an optimum combination of materials, shows a maximum external quantum efficiency of over 12% without using phosphorescent emitters. PMID:25664428

  12. Evaluation of slide based cytometry (SBC) for concentration measurements of fluorescent dyes in solution

    NASA Astrophysics Data System (ADS)

    Pierzchalski, Arkadiusz; Marecka, Monika; Mller, Hans-Willy; Bocsi, Jzsef; Trnok, Attila

    2009-02-01

    Flow cytometers (FCM) are built for particle measurements. In principle, concentration measurement of a homogeneous solution is not possible with FCM due to the lack of a trigger signal. In contrast to FCM slide based cytometry systems could act as tools for the measurement of concentrations using volume defined cell counting chambers. These chambers enable to analyze a well defined volume. Sensovation AG (Stockach, Germany) introduced an automated imaging system that combines imaging with cytometric features analysis. Aim of this study was to apply this imaging system to quantify the fluorescent molecule concentrations. The Lumisens (Sensovation AG) slide-based technology based on fluorescence digital imaging microscopy was used. The instrument is equipped with an inverted microscope, blue and red LEDs, double band-pass filters and a high-resolution cooled 16-bit digital camera. The instrument was focussed on the bottom of 400?m deep 6 chamber slides (IBIDI GmbH, Martinsried, Germany) or flat bottom 96 well plates (Greiner Bio One GmbH, Frickenhausen, Germany). Fluorescent solutions were imaged under 90% pixel saturation in a broad concentration range (FITC: 0.0002-250 ?g/ml, methylene blue (MethB): 0.0002-250 ?g/ml). Exposition times were recorded. Images were analysed by the iCys (CompuCyte Corp., Cambridge, MA, USA) image analysis software with the phantom contour function. Relative fluorescence intensities were calculated from mean fluorescence intensities per phantom contours divided by the exposition time. Solution concentrations could be distinguished over a broad dynamic range of 3.5 to 5.5 decades log (range FITC: 0.0002-31.25?g/ml, MethB: 0.0076-31.25?g/ml) with a good linear relationship between dye concentration and relative fluorescence intensity. The minimal number of fluorescent molecules per pixel as determined by the mean fluorescence intensity and the molecular weight of the fluorochrome were about 800 molecules FITC and ~2.000 MethB. The novel slide-based imaging system is suitable for detection of fluorescence differences over a broad range of concentrations. This approach may lead to novel assays for measuring concentration differences in cell free solutions and cell cultures e.g. in secretion assays.

  13. Reductive fluorescence quenching of the photoexcited free base meso-tetrakis (pentafluorophenyl) porphyrin by amines.

    PubMed

    Prashanthi, Suthari; Kumar, P Hemant; Wang, Li; Perepogu, Arun Kumar; Bangal, Prakriti Ranjan

    2010-03-01

    Steady state and time resolved fluorescence quenching behaviors of meso-Tetrakis (pentafluorophenyl) porphyrin (H(2)F(20)TPP) in presence of different aliphatic and aromatic amines have been executed in homogeneous dichloromethane (DCM) solution. At room temperature in DCM, free base (H(2)F(20)TPP) shows fluorescence with two distinct peaks at 640 and 711 nm and natural lifetime tauf=9.8 ns which are very similar to that of meso-tetraphenyl porphyrin (TPP). Unlike TPP, addition of both aliphatic and aromatic amines to a solution containing H(2)F(20)TPP results in an efficient decrease in fluorescence intensity without altering the shape and peak position of fluorescence emission. Upon addition of amines there was no change in optical absorption spectra of H(2)F(20)TPP. The fluorescence quenching rate constants ranged from 1 x 10(9) to 4 x 10(9) s(-1), which are one order below to the diffusion control limit, and temperature dependent quenching rate constants yield the activation energies which are found to be order of 0.1 eV. Femto second transient absorption studies reveal the existence of amine cation radical and porphyrin anion radicals with very short decay time (15 ps). The fluorescence quenching reaction follows Stern-Volmer kinetics. Steady state and time-resolved data are interpreted within general kinetic scheme of Marcus semi-classical model which attributes bimolecular electron transfer process between amines and the lowest excited singlet state of H(2)F(20)TPP. Calculated internal reorganization energies are found to be in between 0.04 and 0.22 ev. Variation of electron transfer rate as function of free energy change (DeltaG(0)) points the ET reactions in the present systems are in Marcus normal region. This is the first example of reductive fluorescence quenching of free base neutral porphyrins in homogeneous organic solvent ever known. PMID:20063117

  14. A "turn-on" fluorescent sensor for ultrasensitive detection of melamine based on a new fluorescence probe and AuNPs.

    PubMed

    Lu, Qiujun; Zhao, Jiangna; Xue, Shanyan; Yin, Peng; Zhang, Youyu; Yao, Shouzhuo

    2015-02-21

    In this study, we synthesized a new fluorescence probe which was used to detect melamine by coupling with gold nanoparticles (AuNPs). The new fluorescence probe has good optical stability and high fluorescence intensity, which can greatly improve the detection sensitivity. Compared to the traditional fluorophore, it is less dependent on the pH value. It has a very strong fluorescence emission peak at 550 nm, which has larger overlap with the absorption peak of AuNPs. When the probe incubates with the AuNPs, the fluorescence of the probe can be effectively quenched by AuNPs. Adding melamine into a probe-AuNPs mixture caused aggregation of AuNPs and released the adsorbed probe; the fluorescence intensity of the probe was recovered. By measuring the changes of the fluorescence intensity of the probe, the detection of melamine can be realized. Under optimized conditions, the linear response to melamine is in the range of 1.0 10(-8)-4.0 10(-6) mol L(-1) and lowers the detection limit down to 3.0 nmol L(-1) with the sensor. This method can detect melamine in milk and milk-based productions. PMID:25512948

  15. DNA methyltransferase activity detection based on fluorescent silver nanocluster hairpin-shaped DNA probe with 5'-C-rich/G-rich-3' tails.

    PubMed

    Liu, Wenting; Lai, Han; Huang, Rong; Zhao, Chuntao; Wang, Yimo; Weng, Xiaocheng; Zhou, Xiang

    2015-06-15

    DNA methylation has received a large amount of attention due to its close relationship to a wide range of biological phenomena, such as gene activation, gene imprinting, and chromatin stability. Herein, we have designed a hairpin-shaped DNA probe with 5'-C-rich/G-rich-3' tails and developed a simple and reliable fluorescence turn-off assay for DNA adenine methylation (Dam) methyltransferase (MTase) detection combining site recognition and the fluorescence enhancement of DNA-templated silver nanoclusters (DNA-AgNCs) by guanine-rich DNA sequences. A designed hairpin probe with 5' CCCTTACCCC and 3' GGGTGGGGTGGGGTGGGG displays a bright red emission after reacting with AgNO3 and NaBH4. In the presence of Dam MTase, the methylation-sensitive restriction endonuclease Dpn I which has the same recognition site with the Dam MTase can split the probe, freeing the G-rich sequence from the C-rich sequence, thus quenching the fluorescence of DNA-AgNCs. Compared to traditional fluorescent-based methods, this strategy is simple and inexpensive. A linear response to concentrations of Dam MTase which range from 1 U/mL to 100 U/mL and a detection limit of 1 U/mL are obtained without any amplification steps. In addition, we also demonstrate the method can be used for evaluation and screening of inhibitors for Dam MTase. PMID:25682501

  16. "Turn-off-on" fluorescent sensor for (N-methyl-4-pyridyl) porphyrin -DNA and G-quadruplex interactions based on ZnCdSe quantum dots.

    PubMed

    Zhao, Dan; Fan, Yao; Gao, Fang; Yang, Tian-ming

    2015-08-12

    As a new detection model, the reversible fluorescence "turn-off-on" sensor based on quantum dots (QDs) has already been successfully employed in the detections of many biochemical materials, especially in the researches on the interactions between anticancer drugs. The previous studies, however, mainly focused on simple-structured oligonucleotides and Calf thymus DNA. G-quadruplex, an important target for anti-cancer drug with special secondary structure, has been stimulating increasing research interests. In this paper, we report a new detection method based on the fluorescence "turn-off-on" model with water-soluble ZnCdSe QDs as the fluorescent probe, to analyze the interactions between anticancer drug (N-methyl-4-pyridyl) porphyrin (TMPyP) and nucleic acid, especially the G-quadruplex. The fluorescence of QDs can be quenched by TMPyP via photo-induced electron transfer and fluorescence resonance energy transfer, while on the other hand, the combination between TMPyP and G-quadruplex releases QDs from their quenchers and thus recovers the fluorescence. Most importantly, the fluorescence "turn-off-on" model has been employed, for the first time, to analyze the impacts of special factors on the interaction between TMPyP and G-quadruplex. The excellent selectivity of the system has been verified in the studies of the interactions between TMPyP and different DNAs (double-stranded DNA, single-stranded G-quadruplex, and different types of G-quadruplexes) in Na(+) or K(+)-containing buffer. PMID:26320968

  17. Time resolved laser induced fluorescence measurements: Considerations when using Nd:YAG based system

    NASA Astrophysics Data System (ADS)

    Rabasovic, Maja S.; Sevic, Dragutin; Terzic, Mira; Marinkovic, Bratislav P.

    2012-05-01

    Time-resolved laser-induced fluorescence (TR-LIF) and the laser induced breakdown spectroscopy (LIBS) have been shown to be methods which are fast and sensitive to provide information about the constituents in analyzed samples. TR-LIF and LIBS have similar hardware requirements. In this paper, we analyze some characteristics of TR-LIF/LIBS system implemented in our laboratory, considering the fact that the excitation part of the system is based on Nd:YAG laser and Optical Parametric Oscillator (OPO). The laser is more than powerful enough (365 mJ at 1064 nm, variable OPO output >5 mJ) for LIBS, but somehow slow (the length of fundamental laser harmonic output pulse is about 5 ns) for fluorescence measurements in our present area of interest, namely plants and food products. Fortunately, the pulse length of tunable OPO output (320-475 nm) is less then 1 ns, so by means of a correct deconvolution procedure it is possible to measure the fluorescence lifetimes in the range as small as a few nanoseconds. The fluorescence detection part of our system is based on picosecond streak camera. Using the fluorescent dyes (Rhodamine B and Fluorescein) ethanol solutions we verified the analyzing capabilities of our TR-LIF system.

  18. Highly selective and sensitive nanoprobes for cyanide based on gold nanoclusters with red fluorescence emission

    NASA Astrophysics Data System (ADS)

    Zhang, Guomei; Qiao, Yunyun; Xu, Ting; Zhang, Caihong; Zhang, Yan; Shi, Lihong; Shuang, Shaomin; Dong, Chuan

    2015-07-01

    We report a novel and environmentally friendly fluorescent probe for detecting the cyanide ion (CN-) using l-amino acid oxidase (LAAOx)-protected Au nanoclusters (LAAOx@AuNCs) with red emission. The fluorescence-based sensing behaviour of LAAOx@AuNCs towards anions was investigated in buffered aqueous media. Among the anions studied, CN- was found to effectively quench the fluorescence emission of AuNCs based on CN- induced Au core decomposition. Excellent sensitivity and selectivity toward the detection of CN- in aqueous solution were observed. The CN- detection limit was determined to be approximately 180 nM, which is 15 times lower than the maximum level (2700 nM) of CN- in drinking water permitted by the World Health Organization (WHO). A linear relationship between the fluorescence intensity and CN- concentration was observed in two ranges of CN- concentration, including 3.2 10-6 to 3.4 10-5 mol L-1 and 3.81 10-5 to 1.04 10-4 mol L-1. The high sensitivity and selectivity to CN- among the 17 types of anions make the AuNCs good candidates for use in fluorescent nanoprobes of CN-.

  19. Development of Ultrasound-switchable Fluorescence Imaging Contrast Agents based on Thermosensitive Polymers and Nanoparticles

    PubMed Central

    Cheng, Bingbing; Wei, Ming-Yuan; Liu, Yuan; Pitta, Harish; Xie, Zhiwei; Hong, Yi; Nguyen, Kytai T.; Yuan, Baohong

    2015-01-01

    In this work we first introduced a recently developed high-resolution, deep-tissue imaging technique, ultrasound-switchable fluorescence (USF). The imaging principles based on two types of USF contrast agents were reviewed. To improve USF imaging techniques further, excellent USF contrast agents were developed based on high-performance thermoresponsive polymers and environment-sensitive fluorophores. Herein, such contrast agents were synthesized and characterized with five key parameters: (1) peak excitation and emission wavelengths (λex and λem), (2) the fluorescence intensity ratio between on and off states (IOn/IOff), (3) the fluorescence lifetime ratio between on and off states (τOn/τOff), (4) the temperature threshold to switch on fluorophores (Tth), and (5) the temperature transition bandwidth (TBW). We mainly investigated fluorescence intensity and lifetime changes of four environment-sensitive dyes [7-(2-Aminoethylamino)-N,N-dimethyl-4-benzofurazansulfonamide (DBD-ED), St633, Sq660, and St700] as a function of temperature, while the dye was attached to poly(N-isopropylacrylamide) linear polymers or encapsulated in nanoparticles. Six fluorescence resonance energy transfer systems were invented in which both the donor (DBD-ED or ST425) and the acceptor (Sq660) were adopted. Our results indicate that three Förster resonance energy transfer systems, where both IOn/IOff and τOn/τOff are larger than 2.5, are promising for application in future surface tissue bioimaging by USF technique. PMID:26052192

  20. Fabrication of an integrated 670nm VCSEL-based sensor for miniaturized fluorescence sensing

    NASA Astrophysics Data System (ADS)

    O'Sullivan, Thomas D.; Munro, Elizabeth; Harris, James S.; Levi, Ofer

    2010-02-01

    Integrated optical semiconductor sensors are a promising technology for both lab-on-a-chip and molecular imaging applications due to their low cost, small size, high sensitivity, and flexible designs. We present the design and fabrication of a GaAs-based monolithically integrated fluorescence sensor incorporating 670nm VCSELs and PIN photodetectors. This is the first integrated, VCSEL-based fluorescence sensor with excitation at a far-red wavelength and is specifically designed for in vivo sensing applications. In addition, we discuss considerations to simultaneously achieve high power VCSELs and low dark current PIN photodetectors required for sensitive fluorescence detection. These fabricated sensors incorporate 670nm VCSELs emitting 2.0mW at room temperature (RT) with adjacent detectors exhibiting RT dark less than 2pA/mm2 (100mV reverse bias). Fluorescence emission filters suitable for transmitting Cy5.5 fluorescent dye emission were integrated with the photodetectors. The sensor detects Cy5.5 molecules in vitro at 5nM concentration with linear response for concentrations up to 25?M. These miniature sensors are suitable for portable diagnostic assays and in vivo rodent studies.

  1. Comparison of classification algorithms based on fluorescence data for the diagnosis of atherosclerosis

    NASA Astrophysics Data System (ADS)

    Yova, Dido M.; Gonis, Helen; Loukas, Spyros; Kassis, Kyriakos A.; Koukoutsis, Elias; Papaodysseus, Constantinos N.

    1996-01-01

    Fluorescence spectroscopy has been reported as a very promising approach for the discrimination between healthy and atherosclerotic arteries, as far as both, the spectral shape and the intensity of the corresponding spectra seem to be useful parameters for the diagnosis, at specific wavelengths. Nevertheless there are some difficulties in the precise diagnosis, mainly between the different categories of atherosclerotic arteries (fibrous, calcified, heavy calcified). These difficulties are based on the one hand on biophysical factors, such as the necessity for the preknowledge of tissue fluorophores or the complexity of tissue optics. On the other hand, different spectral classification algorithms have been used, such as multivariate linear regression, decision plane analysis and Bayesian decision analysis, each one with certain disadvantages. In this work, two different classification algorithms were developed and evaluated. During the first procedure, simple dimensionless functions were formed by the ratio of the intensities at selected wavelengths and the logistic model was used for statistical analysis. Decision surfaces were drawn and it was estimated that the probability of correct classification is 88%. The algorithm correctly discriminates 97% of healthy from diseased samples and 80% of fibrous from calcified coronary arteries. During the second procedure, a proper ratio was selected in the sense that the ratio groups of the populations P1 and P2 might be separated with an essential considerable veracity probability. The separability was confirmed by testing the validity of specific statistical hypotheses. The demonstration has been made by means of the Kolmogorov-Smirnov goodness of fit method. Therefore by applying statistical methods on proper parameters obtained from the specimens spectra, it has been able to automatically classify the arterial specimens into healthy (normal), fibrous, calcified and heavily calcified, with more than 99.9% probability (less than 0.1% confidence interval). The different classifications algorithms are thoroughly discussed and evaluated.

  2. Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay

    PubMed Central

    Xing, Yun-Peng; Liu, Chun; Zhou, Xiao-Hong; Shi, Han-Chang

    2015-01-01

    This work was the first to report that the kanamycin-binding DNA aptamer (5?-TGG GGG TTG AGG CTA AGC CGA-3?) can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNATO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59?nM with a linear working range of 0.1??M to 20??M for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future. PMID:25634469

  3. Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay

    NASA Astrophysics Data System (ADS)

    Xing, Yun-Peng; Liu, Chun; Zhou, Xiao-Hong; Shi, Han-Chang

    2015-01-01

    This work was the first to report that the kanamycin-binding DNA aptamer (5'-TGG GGG TTG AGG CTA AGC CGA-3') can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNA-TO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59 nM with a linear working range of 0.1 ?M to 20 ?M for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future.

  4. Simultaneous detection and removal of mercury ions in aqueous solution with fluorescent conjugated polymer-based sensor ensemble.

    PubMed

    Kwon, Na Young; Kim, Daigeun; Son, Ji Hye; Jang, Geun Seok; Lee, Jung Hyo; Lee, Taek Seung

    2011-07-15

    A water-soluble, sulfur-containing fluorescent conjugated polymer exhibits a visible fluorescence color change for detection of mercury in the presence of thymine. A new concept provides the design of a sensor ensemble using a simple combination method. This strategy avoids the need for complicated design and synthesis of a recognition group, eliminating the tedious synthetic efforts for the preparation of a sensor material. PMID:21661074

  5. Local SIMPLE multi-atlas-based segmentation applied to lung lobe detection on chest CT

    NASA Astrophysics Data System (ADS)

    Agarwal, M.; Hendriks, E. A.; Stoel, B. C.; Bakker, M. E.; Reiber, J. H. C.; Staring, M.

    2012-02-01

    For multi atlas-based segmentation approaches, a segmentation fusion scheme which considers local performance measures may be more accurate than a method which uses a global performance measure. We improve upon an existing segmentation fusion method called SIMPLE and extend it to be localized and suitable for multi-labeled segmentations. We demonstrate the algorithm performance on 23 CT scans of COPD patients using a leave-one- out experiment. Our algorithm performs significantly better (p < 0.01) than majority voting, STAPLE, and SIMPLE, with a median overlap of the fissure of 0.45, 0.48, 0.55 and 0.6 for majority voting, STAPLE, SIMPLE, and the proposed algorithm, respectively.

  6. Coprecipitative preconcentration and x-ray fluorescence determination of rare earths based on characteristic K-radiation

    SciTech Connect

    Bhagavathy, V.; Sai, P.S.T.; Prasada Rao, T.; Damodaran, A.D.

    1989-01-01

    This paper describes a method for the determination of rare earths at ppb level by coprecipitative preconcentration using iron (III) hydroxide as collector. The precipitates are collected by vacuum filtration onto filter paper containing ethyl cellulose powder. After drying the residue was powdered and pressed into pellets for quantitation by energy dispersive x-ray fluorescence (EDXRF) via their characteristic K-x ray lines. Various parameters that influence coprecipitative preconcentration of rare earths on iron (III) hydroxide were optimized. Further, a numerical method based on simple least square procedure was developed for smoothing and differentiation of the EDXRF data which had been digitized and averaged. By carrying out the least square calculations on a computer via convolution of the EDXRF data (obtained after coprecipitative preconcentration) with properly chosen integers facilitates the determination of as low as 10 ppb each of rare earths.

  7. A reversible DNA-silver nanoclusters-based molecular fluorescence switch and its use for logic gate operation.

    PubMed

    Huang, Zhenzhen; Ren, Jinsong; Qu, Xiaogang

    2012-03-01

    Molecule-like silver nanoclusters (AgNCs) with few to tens of atoms are highly sensitive to the sequence and structure of DNA stabilizers. In this paper, a novel pH-triggered reversible molecular fluorescence switch is developed by taking advantage of the DNA-dependent fluorescence pH response of AgNCs. The DNA-AgNCs fluorescence switch simultaneously addresses concerns of simple construction strategy, efficient design and organic-solvent-free operation. Moreover, the excellent photostability and biocompatibility of AgNCs provide great potential for application of the DNA-AgNCs fluorescence switch in the development of functional molecular devices. Specifically, we apply the DNA-AgNCs fluorescence switch combined with the DNA sequence-dependent pH response pattern of AgNCs for construction of molecular logic gates. PMID:22286835

  8. Synthesis of polymeric fluorescent brightener based on coumarin and its performances on paper as light stabilizer, fluorescent brightener and surface sizing agent

    NASA Astrophysics Data System (ADS)

    Zhang, Guanghua; Zheng, Hua; Guo, Mingyuan; Du, Lun; Liu, Guojun; Wang, Peng

    2016-03-01

    In this work, a novel polymeric fluorescent brightener based on coumarin (PFBC) was synthesized, using three-step synthetic route, from 7-amino-4-methylcoumarin, coumarin monomer (FBC), Acrylamide (AM) and methacrylatoethyl trimethyl ammonium chloride (DMC). The structure of PFBC was characterized by FT-IR, 1HNMR and GPC. PFBC was applied to paper fiber as light stabilizer, fluorescent brightener and surface sizing agent and its performances were evaluated by measuring the UV-vis, fluorescence, thermal stability, the cationic degree, surface strength and smoothness of paper, the brightness degree of paper and the PC value of paper. Results showed that PFBC had better solubility in water than that of FBC, by measuring the optical properties. Through the surface sizing experiment and UV aging experiment, PFBC not only enhanced the surface strength and smoothness of paper as a surface sizing agent, but also had better effect on anti-UV aging than that of FBC as light stabilizer and fluorescent brightener.

  9. A distance-dependent metal-enhanced fluorescence sensing platform based on molecular beacon design.

    PubMed

    Zhou, Zhenpeng; Huang, Hongduan; Chen, Yang; Liu, Feng; Huang, Cheng Zhi; Li, Na

    2014-02-15

    A new metal-enhanced fluorescence (MEF) based platform was developed on the basis of distance-dependent fluorescence quenching-enhancement effect, which combined the easiness of Ag-thiol chemistry with the MEF property of noble-metal structures as well as the molecular beacon design. For the given sized AgNPs, the fluorescence enhancement factor was found to increase with a d(6) dependency in agreement with fluorescence resonance energy transfer mechanism at shorter distance and decrease with a d(-3) dependency in agreement with plasmonic enhancement mechanism at longer distance between the fluorophore and the AgNP surface. As a proof of concept, the platform was demonstrated by a sensitive detection of mercuric ions, using thymine-containing molecular beacon to tune silver nanoparticle (AgNP)-enhanced fluorescence. Mercuric ions were detected via formation of a thymine-mercuric-thymine structure to open the hairpin, facilitating fluorescence recovery and AgNP enhancement to yield a limit of detection of 1 nM, which is well below the U.S. Environmental Protection Agency regulation of the Maximum Contaminant Level Goal (10nM) in drinking water. Since the AgNP functioned as not only a quencher to reduce the reagent blank signal but also an enhancement substrate to increase fluorescence of the open hairpin when target mercuric ions were present, the quenching-enhancement strategy can greatly improve the detection sensitivity and can in principle be a universal approach for various targets when combined with molecular beacon design. PMID:24080216

  10. Nucleic acid based fluorescent sensor for mercury detection

    DOEpatents

    Lu, Yi; Liu, Juewen

    2013-02-05

    A nucleic acid enzyme comprises an oligonucleotide containing thymine bases. The nucleic acid enzyme is dependent on both Hg.sup.2+and a second ion as cofactors, to produce a product from a substrate. The substrate comprises a ribonucleotide, a deoxyribonucleotide, or both.

  11. A turn-off fluorescent biosensor for the rapid and sensitive detection of uranyl ion based on molybdenum disulfide nanosheets and specific DNAzyme

    NASA Astrophysics Data System (ADS)

    Zhang, HongYan; Ruan, YaJuan; Lin, Ling; Lin, Minggui; Zeng, Xiaoxue; Xi, Zhiming; Fu, FengFu

    2015-07-01

    A novel fluorescent biosensor for detecting uranyl ion (UO22+) in aqueous environment has been developed based on the specific recognition of DNAzyme and the fluorescence quenching ability of molybdenum disulfide (MoS2) nanosheets. The DNAzyme contains a DNA enzyme strand and a 6-carboxylfluorescein (FAM)-labeled DNA substrate strand. We demonstrated that MoS2 nanosheets have low affinity to the substrate-enzyme complex DNAzyme. Whereas, in the presence of UO22+, UO22+ can specifically cleave DNAzyme to release FAM-labeled single-strand DNA and the released FAM-labeled single-strand DNA can be firmly adsorbed on the surface of MoS2 nanosheets, which resulted in an obvious decrease of fluorescence intensity. This provided a sensing platform for the rapid, simple and sensitive fluorescent detection of UO22+. By using the sensing platform, a sensitive and selective fluorescent method for the rapid detection of UO22+ has been developed. In comparison with previous biosensor, the proposed method has obvious analytical advantage such as relatively high sensitivity and good stability, short analytical time and low cost. It can be used to detect as low as 2.14 nM of UO22+ in aqueous environment with a recovery of 96-102% and a RSD < 5% (n = 6). The success of this study provides a promising alternative for the rapid and on-site detection of UO22+ in environmental monitoring.

  12. A fluorescent, photochromic and thermochromic trifunctional material based on a layered metal-viologen complex.

    PubMed

    Wan, Fang; Qiu, Li-Xia; Zhou, Liang-Liang; Sun, Yan-Qiong; You, Yi

    2015-11-14

    The azide anion as an energy acceptor and an electron donor has been introduced into a metal-viologen compound to form a 2D layered viologen-based trifunctional material, which exhibits the rare discolored function of reversible photochromism and thermochromism. Interestingly, its fluorescence can be switched by visible light irradiation and heating in air. PMID:26445888

  13. A Palette of Fluorescent Thiophene-Based Ligands for the Identification of Protein Aggregates.

    PubMed

    Shirani, Hamid; Linares, Mathieu; Sigurdson, Christina J; Lindgren, Mikael; Norman, Patrick; Nilsson, K Peter R

    2015-10-19

    By replacing the central thiophene unit of an anionic pentameric oligothiophene with other heterocyclic moities, a palette of pentameric thiophene-based ligands with distinct fluorescent properties were synthesized. All ligands displayed superior selectivity towards recombinant amyloid fibrils as well as disease-associated protein aggregates in tissue sections. PMID:26388448

  14. A Palette of Fluorescent Thiophene-Based Ligands for the Identification of Protein Aggregates

    PubMed Central

    Shirani, Hamid; Linares, Mathieu; Sigurdson, Christina J; Lindgren, Mikael; Norman, Patrick; Nilsson, K Peter R

    2015-01-01

    By replacing the central thiophene unit of an anionic pentameric oligothiophene with other heterocyclic moities, a palette of pentameric thiophene-based ligands with distinct fluorescent properties were synthesized. All ligands displayed superior selectivity towards recombinant amyloid fibrils as well as disease-associated protein aggregates in tissue sections. PMID:26388448

  15. Evaluation of Sustained BMP-2 Release Profiles Using a Novel Fluorescence-Based Retention Assay

    PubMed Central

    Jang, Jun-Hyeog

    2015-01-01

    The purpose of this study was to develop and characterize a novel fluorescence-based retention assay for the evaluation of the release profile of bone morphogenetic protein-2 (BMP-2) released from bone graft carrier. In this study, we evaluated the binding, release kinetics, and delivery efficacies of BMP-2 incorporated into hydroxyapatite (HA) bone grafts. The evaluation of the release profile of BMP-2 from HA bone grafts using a fluorescence-based retention assay revealed initial burst releases from the HA bone grafts followed by long sustained releases up to 14 weeks. The sustained biological activity of the released BMP-2 from HA bone grafts over the full 14-week period supports a long sustained mechanism via fluorescence-based retention assay. Thus, the results from this study show that BMP-2 could be incorporated into HA bone grafts for sustained release over a prolonged period of time with retention of bioactivity and our fluorescence-based retention assay, which is principally detecting the retention profile of BMP-2 in HA bone grafts, is more accurate than conventionally collecting the released BMP-2 for evaluation of BMP-2 release profiles. PMID:25901352

  16. A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase

    PubMed Central

    2015-01-01

    A fluorescent reagentless biosensor for ATP has been developed, based on malonyl-coenzyme A synthetase from Rhodopseudomonas palustris as the protein scaffold and recognition element. Two 5-iodoacetamidotetramethylrhodamines were covalently bound to this protein to provide the readout. This adduct couples ATP binding to a 3.7-fold increase in fluorescence intensity with excitation at 553 nm and emission at 575 nm. It measures ATP concentrations with micromolar sensitivity and is highly selective for ATP relative to ADP. Its ability to monitor enzymatic ATP production or depletion was demonstrated in steady-state kinetic assays in which ATP is a product or substrate, respectively. PMID:26355992

  17. New approach to breast tumor detection based on fluorescence x-ray analysis

    PubMed Central

    Hayashi, Yasuhiko; Okuyama, Fumio

    2010-01-01

    A new technical approach to breast-tumor detection is proposed. The technique is based on fluorescence x-ray analysis, and can identify a miniature malignant tumor within the breast. The primary beam intensity needed in fluorescence x-ray analysis is on a lower order of magnitude than that used in mammography. Thus, the newly-proposed technique would enable detection of a still tiny breast cancer while dramatically lowering the radiation dose. Field-emission x-ray sources might be a key for translating this concept into a medical technique. PMID:20930932

  18. Selective Chemical Labeling of Proteins with Small Fluorescent Molecules Based on Metal-Chelation Methodology

    PubMed Central

    Soh, Nobuaki

    2008-01-01

    Site-specific chemical labeling utilizing small fluorescent molecules is a powerful and attractive technique for in vivo and in vitro analysis of cellular proteins, which can circumvent some problems in genetic encoding labeling by large fluorescent proteins. In particular, affinity labeling based on metal-chelation, advantageous due to the high selectivity/simplicity and the small tag-size, is promising, as well as enzymatic covalent labeling, thereby a variety of novel methods have been studied in recent years. This review describes the advances in chemical labeling of proteins, especially highlighting the metal-chelation methodology.

  19. Fluorescent color factor calculation using dBASE-II.

    PubMed

    King, R L; Carter, H A; Birckbichler, P J

    1986-06-01

    A software system utilizing dBASE-II operating on a dual-drive Apple II+ computer is described. Color factors and retention times for 15 amino acids and epsilon-(gamma-glutamyl)lysine dipeptide are calculated following high performance liquid chromatography. The software package produces a listing of acceptable limits for these parameters calculated as plus and minus 2 standard deviations of the mean. The code is distributed in source form. PMID:3450360

  20. A fluorescence-based method for rapid and direct determination of polybrominated diphenyl ethers in water

    SciTech Connect

    Shan, Huimei; Liu, Chongxuan; Wang, Zheming; Ma, Teng; Shang, Jianying; Pan, Duoqiang

    2015-01-01

    A new method was developed for rapid and direct measurement of polybrominated diphenyl ethers (PBDEs) in aqueous samples using fluorescence spectroscopy. The fluorescence spectra of tri- to deca-BDE (BDE 28, 47, 99, 153, 190, and 209) commonly found in environment were measured at variable emission and excitation wavelengths. The results revealed that the PBDEs have distinct fluorescence spectral profiles and peak positions that can be exploited to identify these species and determine their concentrations in aqueous solutions. The detection limits as determined in deionized water spiked with PBDEs are 1.71-5.82 ng/L for BDE 28, BDE 47, BDE 190, and BDE 209 and 45.55–69.95 ng/L for BDE 99 and BDE 153. The effects of environmental variables including pH, humic substance, and groundwater chemical composition on PBDEs measurements were also investigated. These environmental variables affected fluorescence intensity, but their effect can be corrected through linear additivity and separation of spectral signal contribution. Compared with conventional GC-based analytical methods, the fluorescence spectroscopy method is more efficient as it only uses a small amount of samples (2-4 mL), avoids lengthy complicated concentration and extraction steps, and has a low detection limit of a few ng/L.

  1. Multicolor-based discrimination of 21 short tandem repeats and amelogenin using four fluorescent universal primers.

    PubMed

    Asari, Masaru; Okuda, Katsuhiro; Hoshina, Chisato; Omura, Tomohiro; Tasaki, Yoshikazu; Shiono, Hiroshi; Matsubara, Kazuo; Shimizu, Keiko

    2016-02-01

    The aim of this study was to develop a cost-effective genotyping method using high-quality DNA for human identification. A total of 21 short tandem repeats (STRs) and amelogenin were selected, and fluorescent fragments at 22loci were simultaneously amplified in a single-tube reaction using locus-specific primers with 24-base universal tails and four fluorescent universal primers. Several nucleotide substitutions in universal tails and fluorescent universal primers enabled the detection of specific fluorescent fragments from the 22loci. Multiplex polymerase chain reaction (PCR) produced intense FAM-, VIC-, NED-, and PET-labeled fragments ranging from 90 to 400bp, and these fragments were discriminated using standard capillary electrophoretic analysis. The selected 22loci were also analyzed using two commercial kits (the AmpFLSTR Identifiler Kit and the PowerPlex ESX 17 System), and results for two loci (D19S433 and D16S539) were discordant between these kits due to mutations at the primer binding sites. All genotypes from the 100 samples were determined using 2.5ng of DNA by our method, and the expected alleles were completely recovered. Multiplex 22-locus genotyping using four fluorescent universal primers effectively reduces the costs to less than 20% of genotyping using commercial kits, and our method would be useful to detect silent alleles from commercial kit analysis. PMID:26505528

  2. A fluorescence-based bioassay for antibacterials and its application in screening natural product extracts.

    PubMed

    Michels, Katharina; Heinke, Ramona; Schne, Pia; Kuipers, Oscar P; Arnold, Norbert; Wessjohann, Ludger A

    2015-12-01

    The reliable assessment of the biological activity of a minor component embedded in a complex matrix of several hundred compounds is a difficult but common task in the search for natural product-based antibiotics, for example, by bioassay-guided fractionation. To quantify the antibiotic properties, it is necessary to assess the cell viability. Direct measurements use CFU counts, OD measurements or detection via fluorescent or reducible dyes. However, natural extracts often already possess intrinsic dye, fluorescent, reducing or protein denaturing properties, or they contain insoluble compounds or general protein-binding (tanning) polyphenols as disturbing features, while at the same time very little of the selective antibiotic sought after is present. A promising alternative is provided by intrinsically produced bright fluorescent proteins. In this paper, a rapid, robust and concentration-dependent assay for screening antibiotics with genetically modified mutants of Bacillus subtilis 168 (PabrB-iyfp) is presented. The Gram-positive bacteria exhibit a native fluorescence during their exponential growth phase due to the expression of improved yellow fluorescent protein. To demonstrate the applicability in the field of natural product research, several compounds and extracts were screened for antibacterial activity, with an emphasis on those from the fungal genus Hygrophorus (waxy caps). PMID:26152282

  3. Fluorescent reversible regulation based on the interactions of topotecan hydrochloride, neutral red and quantum dots.

    PubMed

    Wang, Linlin; Shen, Yizhong; Liu, Shaopu; Yang, Jidong; Liang, Wanjun; Li, Dan; He, Youqiu

    2014-10-24

    The interactions of topotecan hydrochloride (THC), neutral red (NR) and thioglycolic acid (TGA) capped CdTe/CdS quantum dots (QDs) built a solid base for the controlling of the fluorescent reversible regulation of the system. This study was developed by means of ultraviolet-visible (UV-vis) absorption, fluorescence (FL), resonance Rayleigh scattering (RRS) spectroscopy and transmission electron microscopy (TEM). Corresponding experimental results revealed that the fluorescence of TGA-CdTe/CdS QDs could be effectively quenched by NR, while the RRS of the QDs enhanced gradually with the each increment of NR concentration. After the addition of THC, the strong covalent conjugation between NR and THC which was in carboxylate state enabled NR to be dissociated from the surface of TGA-CdTe/CdS QDs to form more stable complex with THC, thereby enhancing the fluorescence of the TGA-CdTe/CdS QDs-NR system. What is more, through analyzing the optical properties and experimental data of the reaction between TGA-CdTe/CdS QDs and NR, the possible reaction mechanism of the whole system was discussed. This combination of multiple spectroscopic techniques could contribute to the investigation for the fluorescent reversible regulation of QDs and a method could also be established to research the interactions between camptothecin drugs and dyes. PMID:25459722

  4. A Fluorescence-Based Method for Rapid and Direct Determination of Polybrominated Diphenyl Ethers in Water

    PubMed Central

    Shan, Huimei; Ma, Teng; Shang, Jianying; Pan, Duoqiang

    2015-01-01

    A new method was developed for rapid and direct measurement of polybrominated diphenyl ethers (PBDEs) in aqueous samples using fluorescence spectroscopy. The fluorescence spectra of tri- to deca-BDE (BDE 28, 47, 99, 153, 190, and 209) commonly found in environment were measured at variable emission and excitation wavelengths. The results revealed that the PBDEs have distinct fluorescence spectral profiles and peak positions that can be exploited to identify these species and determine their concentrations in aqueous solutions. The detection limits as determined in deionized water spiked with PBDEs are 1.71–5.82 ng/L for BDE 28, BDE 47, BDE 190, and BDE 209 and 45.55–69.95 ng/L for BDE 99 and BDE 153. The effects of environmental variables including pH, humic substance, and groundwater chemical composition on PBDEs measurements were also investigated. These environmental variables affected fluorescence intensity, but their effect can be corrected through linear additivity and separation of spectral signal contribution. Compared with conventional GC-based analytical methods, the fluorescence spectroscopy method is more efficient as it only uses a small amount of samples (2–4 mL), avoids lengthy complicated concentration and extraction steps, and has a low detection limit of a few ng/L. PMID:25705548

  5. 2D fluorescence spectroscopy for monitoring ion-exchange membrane based technologies - Reverse electrodialysis (RED).

    PubMed

    Pawlowski, Sylwin; Galinha, Claudia F; Crespo, João G; Velizarov, Svetlozar

    2016-01-01

    Reverse electrodialysis (RED) is one of the emerging, membrane-based technologies for harvesting salinity gradient energy. In RED process, fouling is an undesirable operation constraint since it leads to a decrease of the obtainable net power density due to increasing stack electric resistance and pressure drop. Therefore, early fouling detection is one of the main challenges for successful RED technology implementation. In the present study, two-dimensional (2D) fluorescence spectroscopy was used, for the first time, as a tool for fouling monitoring in RED. Fluorescence excitation-emission matrices (EEMs) of ion-exchange membrane surfaces and of natural aqueous streams were acquired during one month of a RED stack operation. Fouling evolvement on the ion-exchange membrane surfaces was successfully followed by 2D fluorescence spectroscopy and quantified using principal components analysis (PCA). Additionally, the efficiency of cleaning strategy was assessed by measuring the membrane fluorescence emission intensity before and after cleaning. The anion-exchange membrane (AEM) surface in contact with river water showed to be significantly affected due to fouling by humic compounds, which were found to cross through the membrane from the lower salinity (river water) to higher salinity (sea water) stream. The results obtained show that the combined approach of using 2D fluorescence spectroscopy and PCA has a high potential for studying fouling development and membrane cleaning efficiency in ion exchange membrane processes. PMID:26497936

  6. Fluorescent reversible regulation based on the interactions of topotecan hydrochloride, neutral red and quantum dots

    NASA Astrophysics Data System (ADS)

    Wang, Linlin; Shen, Yizhong; Liu, Shaopu; Yang, Jidong; Liang, Wanjun; Li, Dan; He, Youqiu

    2015-02-01

    The interactions of topotecan hydrochloride (THC), neutral red (NR) and thioglycolic acid (TGA) capped CdTe/CdS quantum dots (QDs) built a solid base for the controlling of the fluorescent reversible regulation of the system. This study was developed by means of ultraviolet-visible (UV-vis) absorption, fluorescence (FL), resonance Rayleigh scattering (RRS) spectroscopy and transmission electron microscopy (TEM). Corresponding experimental results revealed that the fluorescence of TGA-CdTe/CdS QDs could be effectively quenched by NR, while the RRS of the QDs enhanced gradually with the each increment of NR concentration. After the addition of THC, the strong covalent conjugation between NR and THC which was in carboxylate state enabled NR to be dissociated from the surface of TGA-CdTe/CdS QDs to form more stable complex with THC, thereby enhancing the fluorescence of the TGA-CdTe/CdS QDs-NR system. What is more, through analyzing the optical properties and experimental data of the reaction between TGA-CdTe/CdS QDs and NR, the possible reaction mechanism of the whole system was discussed. This combination of multiple spectroscopic techniques could contribute to the investigation for the fluorescent reversible regulation of QDs and a method could also be established to research the interactions between camptothecin drugs and dyes.

  7. Enzymatic cascade based fluorescent DNAzyme machines for the ultrasensitive detection of Cu(II) ions.

    PubMed

    He, Jing-Lin; Zhu, Shuang-Li; Wu, Ping; Li, Pan-Pan; Li, Ting; Cao, Zhong

    2014-10-15

    A novel enzymatic cascade based fluorescent DNAzyme machine has been developed for the amplified detection of copper (Cu(2+)) ions. This is the first attempt to carry out the combination of the self-cleaving DNAzyme and the polymerase/endonuclease reaction cycles involving cleaved substrate extension. In the presence of Cu(2+) ions, the enzyme strand carries out catalytic reactions to hydrolytic cleavage of the substrate strand. The cleaved DNAzyme substrates act as primers and trigger the Klenow Fragment polymerization. Nb.BbvCI endonuclease cuts the double-stranded niking site and thus opens a new site for a new replication. The replication regenerates the complete dsDNA to initiate another cycle of nicking, polymerization and displacement. Finally the fluorescence dye, SG, inserts into the DNA double helix to generate a distinguishable fluorescence enhancement. The Cu(2+) ions act as the activator for enzymatic cascade amplification generating multiple duplex structures in the nascent product. An increasing fluorescence is observed with increasing Cu(2+) ions concentration. A good nonlinear correlation (R=0.9997) was obtained between fluorescence intensity and the cubic logarithm of the Cu(2+) ions concentration over the range 0.50-200 nM. This nonlinear response phenomenon results in an efficient improvement of the sensitivity of our current proposed assay. The activation of such enzymatic cascades through analyte-DNAzyme interactions is not only valuable to activate the cooperation of enzyme networks, but also has a substantial impact on the development of amplified DNAzyme sensors. PMID:24787125

  8. Fluorescence suppression using wavelength modulated Raman spectroscopy in fiber-probe-based tissue analysis.

    PubMed

    Praveen, Bavishna B; Ashok, Praveen C; Mazilu, Michael; Riches, Andrew; Herrington, Simon; Dholakia, Kishan

    2012-07-01

    In the field of biomedical optics, Raman spectroscopy is a powerful tool for probing the chemical composition of biological samples. In particular, fiber Raman probes play a crucial role for in vivo and ex vivo tissue analysis. However, the high-fluorescence background typically contributed by the auto fluorescence from both a tissue sample and the fiber-probe interferes strongly with the relatively weak Raman signal. Here we demonstrate the implementation of wavelength-modulated Raman spectroscopy (WMRS) to suppress the fluorescence background while analyzing tissues using fiber Raman probes. We have observed a significant signal-to-noise ratio enhancement in the Raman bands of bone tissue, which have a relatively high fluorescence background. Implementation of WMRS in fiber-probe-based bone tissue study yielded usable Raman spectra in a relatively short acquisition time (∼30  s), notably without any special sample preparation stage. Finally, we have validated its capability to suppress fluorescence on other tissue samples such as adipose tissue derived from four different species. PMID:22894519

  9. Development of fluorescence based handheld imaging devices for food safety inspection

    NASA Astrophysics Data System (ADS)

    Lee, Hoyoung; Kim, Moon S.; Chao, Kuanglin; Lefcourt, Alan M.; Chan, Diane E.

    2013-05-01

    For sanitation inspection in food processing environment, fluorescence imaging can be a very useful method because many organic materials reveal unique fluorescence emissions when excited by UV or violet radiation. Although some fluorescence-based automated inspection instrumentation has been developed for food products, there remains a need for devices that can assist on-site inspectors performing visual sanitation inspection of the surfaces of food processing/handling equipment. This paper reports the development of an inexpensive handheld imaging device designed to visualize fluorescence emissions and intended to help detect the presence of fecal contaminants, organic residues, and bacterial biofilms at multispectral fluorescence emission bands. The device consists of a miniature camera, multispectral (interference) filters, and high power LED illumination. With WiFi communication, live inspection images from the device can be displayed on smartphone or tablet devices. This imaging device could be a useful tool for assessing the effectiveness of sanitation procedures and for helping processors to minimize food safety risks or determine potential problem areas. This paper presents the design and development including evaluation and optimization of the hardware components of the imaging devices.

  10. A fluorescence-based method for rapid and direct determination of polybrominated diphenyl ethers in water

    DOE PAGESBeta

    Shan, Huimei; Liu, Chongxuan; Wang, Zheming; Ma, Teng; Shang, Jianying; Pan, Duoqiang

    2015-01-01

    A new method was developed for rapid and direct measurement of polybrominated diphenyl ethers (PBDEs) in aqueous samples using fluorescence spectroscopy. The fluorescence spectra of tri- to deca-BDE (BDE 28, 47, 99, 153, 190, and 209) commonly found in environment were measured at variable emission and excitation wavelengths. The results revealed that the PBDEs have distinct fluorescence spectral profiles and peak positions that can be exploited to identify these species and determine their concentrations in aqueous solutions. The detection limits as determined in deionized water spiked with PBDEs are 1.71-5.82 ng/L for BDE 28, BDE 47, BDE 190, and BDEmore » 209 and 45.55–69.95 ng/L for BDE 99 and BDE 153. The effects of environmental variables including pH, humic substance, and groundwater chemical composition on PBDEs measurements were also investigated. These environmental variables affected fluorescence intensity, but their effect can be corrected through linear additivity and separation of spectral signal contribution. Compared with conventional GC-based analytical methods, the fluorescence spectroscopy method is more efficient as it only uses a small amount of samples (2-4 mL), avoids lengthy complicated concentration and extraction steps, and has a low detection limit of a few ng/L.« less

  11. Anion recognition by simple chromogenic and chromo-fluorogenic salicylidene Schiff base or reduced-Schiff base receptors

    NASA Astrophysics Data System (ADS)

    Dalapati, Sasanka; Jana, Sankar; Guchhait, Nikhil

    2014-08-01

    This review contains extensive application of anion sensing ability of salicylidene type Schiff bases and their reduced forms having various substituents with respect to phenolic sbnd OH group. Some of these molecular systems behave as receptor for recognition or sensing of various anions in organic or aqueous-organic binary solvent mixture as well as in the solid supported test kits. Development of Schiff base or reduced Schiff base receptors for anion recognition event is commonly based on the theory of hydrogen bonding interaction or deprotonation of phenolic -OH group. The process of charge transfer (CT) or inhibition of excited proton transfer (ESIPT) or followed by photo-induced electron transfer (PET) lead to naked-eye color change, UV-vis spectral change, chemical shift in the NMR spectra and fluorescence spectral modifications. In this review we have tried to discuss about the anion sensing properties of Schiff base or reduced Schiff base receptors.

  12. A Fluorescent Thermometer Based on a Pyrene-Labeled Thermoresponsive Polymer

    PubMed Central

    Pietsch, Christian; Vollrath, Antje; Hoogenboom, Richard; Schubert, Ulrich S.

    2010-01-01

    Thermoresponsive polymers that undergo a solubility transition by variation of the temperature are important materials for the development of smart materials. In this contribution we exploit the solubility phase transition of poly(methoxy diethylene glycol methacrylate), which is accompanied by a transition from hydrophilic to hydrophobic, for the development of a fluorescent thermometer. To translate the polymer phase transition into a fluorescent response, the polymer was functionalized with pyrene resulting in a change of the emission based on the microenvironment. This approach led to a soluble polymeric fluorescent thermometer with a temperature range from 11 C to 21 C. The polymer phase transition that occurs during sensing is studied in detail by dynamic light scattering. PMID:22163636

  13. Assessment of fluorescein-based fluorescent dyes for tracing Neotyphodium endophytes in planta.

    PubMed

    Card, Stuart D; Tapper, Brian A; Lloyd-West, Catherine; Wright, Kathryn M

    2013-01-01

    Fluorescent dyes were assessed for their ability to stain viable hyphae of the fungi Neotyphodium lolii and N. coenophialum, symbiotic endophytes of the Pooideae grasses Lolium perenne and Festuca arundinacea, respectively. The fluorescein-based fluorophores; fluorescein diacetate (FDA), 5(6)-carboxy-fluorescein diacetate (CFDA), 5-chloromethylfluorescein diacetate (CMFDA) and the chitin-binding stain, Calcofluor while M2R, were assessed for staining of endophyte hyphae in vitro from axenic fungal cultures and in planta, including epidermal leaf sheath peels, nodes, ovaries, embryos and meristems. CMFDA produced the greatest intensity of staining of fungal hyphae and gave excellent contrast in planta compared to the plant cells. Compared to the other dyes, CMFDA was also the least affected by photo bleaching and continued to fluoresce up to 2 h after initial excitation. None of the fluorescent dyes stained fungal hyphae in seed. PMID:22802389

  14. Classical low-coherence interferometry based on broadband parametric fluorescence and amplification.

    PubMed

    Le Gout, Julien; Venkatraman, Dheera; Wong, Franco N C; Shapiro, Jeffrey H

    2009-09-28

    We demonstrate that single-mode broadband amplified spontaneous parametric downconversion, combined with optical parametric amplification, can be used as a classical source of phase-sensitive cross-correlated beams. We first study the single spatial mode emission and the spectral brightness properties of the parametric fluorescence, produced in periodically poled MgO-doped lithium niobate. Using the same single-pass bulk-crystal configuration for a pulsed optical parametric amplifier, we achieve a gain of approximately 20 dB at an average pump power of 2W, and explain the pulse narrowing observed at the output of both parametric fluorescence and amplification in the regime of high gain. Combining these two nonlinear processes, we measured optical coherence tomography signals with standard InGaAs photodiodes, thus realizing the first classical interferometer based on amplified parametric fluorescence. The results suggest their utility for demonstrating phase-conjugate optical coherence tomography. PMID:19907576

  15. A single-wavelength-emitting ratiometric probe based on phototriggered fluorescence switching of graphene quantum dots.

    PubMed

    Qu, Zhi-bei; Zhang, Min; Zhou, Tianshu; Shi, Guoyue

    2014-10-13

    Ratiometric fluorescent probes are of great importance in research, because a built-in correction for environmental effects can be provided to reduce background interference. However, the traditional ratiometric fluorescent probes require two luminescent materials with different emission bands. Herein a novel ratiometric probe based on a single-wavelength-emitting material is reported. The probe works by regulating the luminescent property of graphene quantum dots with UV illumination as activator. The ratiometric sensor shows high sensitivity and specificity for iron ions. Moreover, the ratiometric sensor was successfully employed to monitor ferritin levels in Sprague Dawley rats with chemical-induced acute liver damage. The proposed single-wavelength ratiometric fluorescent probe may greatly broaden the applicability of ratiometric sensors in diagnostic devices, medical applications, and analytical chemistry. PMID:25187434

  16. A dithienosilole-based fluorescent chemosensor for multiple logic operations at the molecular level.

    PubMed

    Zhang, Chen; Sun, Caixia; Lu, Yahong; Wang, Junxing; He, Xingxing; Lu, Junting; Yin, Shouchun; Qiu, Huayu

    2015-11-01

    A chemosensor consisting of two terpyridines covalently linked to a dithienosilole unit (1) has been synthesized, and its optical and metal sensing properties have been investigated. Due to the metal-organic coordination function, 1 can bind with many transition metal ions and display different fluorescence responses that cause it to function as a "turn-off" fluorescent chemosensor. A significant bathochromic shift in the fluorescence spectra is observed in the presence of Zn(2+). Meanwhile, the emission of 1 is weakened upon exposure to Ag(+) and Fe(2+) and completely quenched by Ni(2+), Co(2+), and Cu(2+). Based on the observed results, several logic gates, such as XNOR, INHIBIT, and IMPLICATION, have been achieved by controlling the chemical inputs. PMID:26099823

  17. Hyper-spectral modulation fluorescent imaging using double acousto-optical tunable filter based on TeO2-crystals

    NASA Astrophysics Data System (ADS)

    Zaytsev, Kirill I.; Perchik, Alexey V.; Chernomyrdin, Nikita V.; Kudrin, Konstantin G.; Reshetov, Igor V.; Yurchenko, Stanislav O.

    2015-01-01

    We have proposed a method for hyper-spectral fluorescent imaging based on acousto-optical filtering. The object of interest was pumped using ultraviolet radiation of mercury lamp equipped with monochromatic excitation filter with the window of transparency centered at 365 nm. Double TeO2-based acousto-optical filter, tunable in range from 430 to 780 nm and having 2 nm bandwidth of spectral transparency, was used in order to detect quasimonochromatic images of object fluorescence. Modulating of ultraviolet pump intensity was used in order to reduce an impact of non-fluorescent background on the sample fluorescent imaging. The technique for signal-to-noise ratio improvement, based on fluorescence intensity estimation via digital processing of modulated video sequence of fluorescent object, was introduced. We have implemented the proposed technique for the test sample studying and we have discussed its possible applications.

  18. Label-Free Detection of Sequence-Specific DNA Based on Fluorescent Silver Nanoclusters-Assisted Surface Plasmon-Enhanced Energy Transfer.

    PubMed

    Ma, Jin-Liang; Yin, Bin-Cheng; Le, Huynh-Nhu; Ye, Bang-Ce

    2015-06-17

    We have developed a label-free method for sequence-specific DNA detection based on surface plasmon enhanced energy transfer (SPEET) process between fluorescent DNA/AgNC string and gold nanoparticles (AuNPs). DNA/AgNC string, prepared by a single-stranded DNA template encoded two emitter-nucleation sequences at its termini and an oligo spacer in the middle, was rationally designed to produce bright fluorescence emission. The proposed method takes advantage of two strategies. The first one is the difference in binding properties of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) toward AuNPs. The second one is SPEET process between fluorescent DNA/AgNC string and AuNPs, in which fluorescent DNA/AgNC string can be spontaneously adsorbed onto the surface of AuNPs and correspondingly AuNPs serve as "nanoquencher" to quench the fluorescence of DNA/AgNC string. In the presence of target DNA, the sensing probe hybridized with target DNA to form duplex DNA, leading to a salt-induced AuNP aggregation and subsequently weakened SPEET process between fluorescent DNA/AgNC string and AuNPs. A red-to-blue color change of AuNPs and a concomitant fluorescence increase were clearly observed in the sensing system, which had a concentration dependent manner with specific DNA. The proposed method achieved a detection limit of ?2.5 nM, offering the following merits of simple design, convenient operation, and low experimental cost because of no chemical modification, organic dye, enzymatic reaction, or separation procedure involved. PMID:26024337

  19. A fluorescent chemosensor for sodium based on photoinduced electron transfer.

    PubMed

    He, Huarui; Mortellaro, Mark A; Leiner, Marc J P; Young, Susanne T; Fraatz, Robert J; Tusa, James K

    2003-02-01

    A new optical sensor suitable for practical measurement of sodium in serum and whole blood samples is described. The optical sensor is based on a novel PET (photoinduced electron transfer) fluoroionophore immobilized in a hydrophilic polymer layer. The design concept of the fluoroionophore follows the receptor-spacer-fluorophore approach to sensor design using intramolecular PET-based signal transduction. Key to the development of this sensor is the identification of a nitrogen-containing, sodium-binding ionophore, coupled with a fluorophore having the correct spectral and electron-accepting properties. The slope of the sensor is approximately 0.5%/mM in the typical clinically significant range of 120-160 mM. This sensor has been implemented into a disposable cartridge, used for a commercially available critical care analyzer (Roche OPTI CCA) with precision better than +/- 1 mM (1 SD). The sensor displays excellent stability against hydrolysis and oxidation, leading to slope changes <5% after 9 months wet storage at 30 degrees C. On the basis of this design concept, fluoroionophores for other cations such as potassium, calcium and magnesium can be prepared by substitution of the ionophore. PMID:12585483

  20. Glass-based fluorescence reference materials used for optical and biophotonic applications

    NASA Astrophysics Data System (ADS)

    Engel, A.; Ottermann, C.; Resch-Genger, U.; Hoffmann, K.; Schweizer, S.; Selling, J.; Spaeth, J.-M.; Rupertus, V.

    2006-04-01

    Fluorescence techniques are known for their high sensitivity and are widely used as analytical tools and detection methods for product and process control, material sciences, environmental and biotechnical analysis, molecular genetics, cell biology, medical diagnostics, and drug screening. For routine measurements by fluorescence techniques the existence of an improved quality assurance is one of the basic needs. According to DIN/ISO 17025 certified standards are used for fluorescence diagnostics having the drawback of giving relative values only. Typical requirements onto fluorescence reference materials or standards deal with the verification of the instrument performance as well as the improvement of the data comparability. Especially for biomedical applications fluorescence labels are used for the detection of proteins. In particular these labels consist of nano crystalline materials like CdS and CdSe. The field of Non-Cadmium containing materials is under investigation. In order to evaluate whether glass based materials can be used as standards it is necessary to calculate absolute values like absorption/excitation cross sections or relative quantum yields. This can be done using different quantities of dopands in glass, glass ceramics or crystals. The investigated materials are based on different types of glass, silicate, phosphate and boron glass, which play a dominant role for the absorption and emission mechanism. Additional to the so-called elementary fluorescence properties induced by raw earth elements the formation of defects lead to higher cross sections additionally. The main investigations deal with wavelength accuracy and lifetime of doped glasses, glass ceramics and crystalline samples. Moreover intensity patterns, homogeneity aspects and photo stability will be discussed.

  1. Paper-based fluorescence resonance energy transfer assay for directly detecting nucleic acids and proteins.

    PubMed

    Li, Hua; Fang, Xueen; Cao, Hongmei; Kong, Jilie

    2016-06-15

    Paper-based fluorescence resonance energy transfer assay (FRET) is gaining great interest in detecting macro-biological molecule. It is difficult to achieve conveniently and fast detection for macro-biological molecule. Herein, a graphene oxide (GO)-based paper chip (glass fiber) integrated with fluorescence labeled single-stranded DNA (ssDNA) for fast, inexpensive and direct detection of biological macromolecules (proteins and nucleic acids) has been developed. In this paper, we employed the Cy3/FAM-labeled ssDNA as the reporter and the GO as quencher and the original glass fiber paper as data acquisition substrates. The chip which was designed and fabricated by a cutting machine is a miniature biosensor that monitors fluorescence recovery from resonance energy transfer. The hybridization assays and fluorescence detection were all simplified, and the surface of the chip did not require immobilization or washing. A Nikon Eclipse was employed as excited resource and a commercial digital camera was employed for capturing digital images. This paper-based microfluidics chip has been applied in the detection of proteins and nucleic acids. The biosensing capability meets many potential requirements for disease diagnosis and biological analysis. PMID:26807518

  2. Fluorescent chemosensor for pyridine based on N-doped carbon dots.

    PubMed

    Campos, B B; Abellán, C; Zougagh, M; Jimenez-Jimenez, J; Rodríguez-Castellón, E; Esteves da Silva, J C G; Ríos, A; Algarra, M

    2015-11-15

    Fluorescent carbon dots (CDs) and its nitrogen doped (N-CDs) nanoparticles have been synthesized from lactose as precursor using a bottom-up hydrothermal methodology. The synthesized nanoparticles have been characterized by elemental analysis, FTIR, Raman, TEM, DLS, XPS, and steady-state and life-time fluorescence. The synthesized carbon nanoparticles, CDs and N-CDs, have a size at about 7.7±2.4 and 50±15nm, respectively, and quantum yields of 8% (CDs) and 11% (N-CDs). These techniques demonstrated the effectiveness of the synthesis procedure and the functionalization of the CDs surface with amine and amide groups in the presence of NH3 in aqueous media. The effect of excitation wavelength and pH on the luminescent properties was studied. Under the optimal conditions, the nitrogen doped nanoparticles can be used as pyridine sensor in aqueous media because they show an enhancement of its fluorescence with a good linear relationship. The analytical method is simple, reproducible and very sensitive for pyridine determination. PMID:26225491

  3. Ratiometric fluorescence detection of silver ions using thioflavin T-based organic/inorganic hybrid supraparticles.

    PubMed

    Li, Yan-Yun; Zhang, Min; Lu, Ling-Fei; Zhu, Anwei; Xia, Fei; Zhou, Tianshu; Shi, Guoyue

    2015-09-01

    In this work, we present a new type of functional organic/inorganic hybrid supraparticle that spontaneously assembles from silver ions (Ag(+)), iodide ions (I(-)) and thioflavin T (ThT) under aqueous solution conditions. ThT alone in aqueous solution was weakly fluorescent with an emission band at 494 nm, which was related to the monomer. However, in the above-mentioned hybrid supraparticle (i.e., ThT@AgI SP) structure, the ThT monomer can form a dimer with a new emission band. The new band shifted to 546 nm and the emission intensity increased. We further present a facile strategy of reversible fluorescence switching of ThT by a simple cation (Ag(+)) and anions (I(-) and S(2-)), which can be employed for the ratiometric fluorescence detection of Ag(+) with high sensitivity and selectivity. The linear range of detecting Ag(+) was from 100 nM to 10 ?M, with a limit of detection as low as approximately 50 nM. Moreover, it can be successfully applied for the operation of a logic gate system and to the sensing of Ag(+) in real water samples. PMID:26212864

  4. Highly sensitive fluorescent immunosensor for detection of influenza virus based on Ag autocatalysis.

    PubMed

    Li, Yanxia; Hong, Mei; Qiu, Bin; Lin, Zhenyu; Chen, Yiting; Cai, Zongwei; Chen, Guonan

    2014-04-15

    A versatile, ultrasensitive immunosensor for detection of influenza virus was designed by combining silver nanoparticles (Ag NPs) labeled antibodies with indirect fluorescence. A new technology using Ag-S covalent binding was applied for antibody labeling. Influenza A (H1N1) virus, as a subtype of influenza A virus that was the most common cause of human influenza (flu), was acted as the target antigen using sandwich type-immunoreactions on the high binding ELISA plates. The antibody-labeled Ag NPs were then released by acid solution to produce Ag(+) which can catalyze o-phenylenediamine (OPDA) oxidation to produce fluorescence for highly sensitive detection. Under the optimal conditions, it shows good linear relationship between fluorescence intensity and the logarithm of the concentration of H1N1 over the range of 1.010(-12)-1.010(-8) g mL(-1) with a detection limit (LOD, 3?) of 1.010(-13) g mL(-1). Results indicated that the proposed method give a good sensitivity and simple operation for detecting the influenza virus. This work also provided a promising potential for antigen detection by Ag NPs labeled, and the steps were easy to handle. PMID:24292140

  5. Fluorescence resonance energy transfer-based ratiometric fluorescent assay for highly sensitive and selective determination of sulfide anions.

    PubMed

    Liang, Meijuan; Chen, Yonglei; Zhang, Haijuan; Niu, Xiaoying; Xu, Laifang; Ren, Cuiling; Chen, Xingguo

    2015-10-01

    A novel and effective ratiometric fluorescence strategy was developed for rapidly, sensitively and selectively probing sulfide anions (S(2-)). A dual-emission nanosensor was prepared by covalently attaching fluorescent carbon nanoparticles (CNPs) to gold nanoclusters (Au NCs), triggering the sensing mechanism of fluorescence resonance energy transfer (FRET) from CNPs (donor) to Au NCs (acceptor). Once S(2-) was added, considerable fluorescence recovery of CNPs and quenching of Au NCs were observed due to the inhibition of FRET progress via the formation of Au2S. The ratiometric probe showed good, specific S(2-) sensing behavior and high sensitivity with a detection limit of 18 nM. Significantly, the assay was successfully employed to determine the S(2-) content in biological and water samples, presenting immense promise in the biological and environmental fields. PMID:26317130

  6. A novel method for image denoising of fluorescence molecular imaging based on fuzzy C-Means clustering

    NASA Astrophysics Data System (ADS)

    An, Yu; Liu, Jie; Ye, Jinzuo; Mao, Yamin; Yang, Xin; Jiang, Shixin; Chi, Chongwei; Tian, Jie

    2015-03-01

    As an important molecular imaging modality, fluorescence molecular imaging (FMI) has the advantages of high sensitivity, low cost and ease of use. By labeling the regions of interest with fluorophore, FMI can noninvasively obtain the distribution of fluorophore in-vivo. However, due to the fact that the spectrum of fluorescence is in the section of the visible light range, there are mass of autofluorescence on the surface of the bio-tissues, which is a major disturbing factor in FMI. Meanwhile, the high-level of dark current for charge-coupled device (CCD) camera and other influencing factor can also produce a lot of background noise. In this paper, a novel method for image denoising of FMI based on fuzzy C-Means clustering (FCM) is proposed, because the fluorescent signal is the major component of the fluorescence images, and the intensity of autofluorescence and other background signals is relatively lower than the fluorescence signal. First, the fluorescence image is smoothed by sliding-neighborhood operations to initially eliminate the noise. Then, the wavelet transform (WLT) is performed on the fluorescence images to obtain the major component of the fluorescent signals. After that, the FCM method is adopt to separate the major component and background of the fluorescence images. Finally, the proposed method was validated using the original data obtained by in vivo implanted fluorophore experiment, and the results show that our proposed method can effectively obtain the fluorescence signal while eliminate the background noise, which could increase the quality of fluorescence images.

  7. A fluorescence detection of D-penicillamine based on Cu2+-induced fluorescence quenching system of protein-stabilized gold nanoclusters

    NASA Astrophysics Data System (ADS)

    Wang, Peng; Li, Bang Lin; Li, Nian Bing; Luo, Hong Qun

    2015-01-01

    In this contribution, a luminescent gold nanoclusters which were synthesized by bovine serum albumin as novel fluorescent probes were successfully utilized for the determination of D-penicillamine for the first time. Cupric ion was employed to quench the strong fluorescence of the gold nanoclusters, whereas the addition of D-penicillamine caused obvious restoration of fluorescence intensity of the Cu2+-gold nanoclusters system. Under optimum conditions, the increment in fluorescence intensity of Cu2+-gold nanoclusters system caused by D-penicillamine was linearly proportional to the concentration of D-penicillamine in the range of 2.0 10-5-2.39 10-4 M. The detection limit for D-penicillamine was 5.4 10-6 M. With the off-on fluorescence signal at 650 nm approaching the near-infrared region, the present sensor for D-penicillamine detection had high sensitivity and low spectral interference. Furthermore, the novel gold nanoclusters-based fluorescent sensor has been applied to the determination of D-penicillamine in real biological samples with satisfactory results.

  8. Development of a novel fluorescence probe capable of assessing the cytoplasmic entry of siderophore-based conjugates.

    PubMed

    Kim, Hyeon Seok; Song, Woon Young; Kim, Hak Joong

    2015-01-01

    A novel fluorescence probe capable of assessing the cytoplasmic entry of siderophore-based conjugates was synthesized and evaluated by photochemical characterization and cell-based assays. The specific responsiveness to the cytoplasmic entry of the probe was implemented by adopting a disulfide linker, whose cleavage under the reducing conditions of the cytoplasm induced the display of a distinctive fluorescence signal. PMID:25374128

  9. Aptamers-based sandwich assay for silver-enhanced fluorescence multiplex detection.

    PubMed

    Wang, Ying; Li, Hui; Xu, Danke

    2016-01-28

    In this work, aptamers-modified silver nanoparticles (AgNPs) were prepared as capture substrate, and fluorescent dyes-modified aptamers were synthesized as detection probes. The sandwich assay was based on dual aptamers, which was aimed to accomplish the highly sensitive detection of single protein and multiplex detection of proteins on one-spot. We found that aptamers-modified AgNPs based microarray was much superior to the aptamer based microarray in fluorescence detection of proteins. The result shows that the detection limit of the sandwich assay using AgNPs probes for thrombin or platelet-derived growth factor-BB (PDGF-BB) is 80 or 8 times lower than that of aptamers used directly. For multiplex detection of proteins, the detection limit was 625pM for PDGF-BB and 21pM for thrombin respectively. The sandwich assay based on dual aptamers and AgNPs was sensitive and specific. PMID:26755149

  10. Fluorescent Aptamer Sensors

    NASA Astrophysics Data System (ADS)

    Chen, Hui William; Kim, Youngmi; Meng, Ling; Mallikaratchy, Prabodhika; Martin, Jennifer; Tang, Zhiwen; Shangguan, Dihua; O'Donoghue, Meghan; Tan, Weihong

    Aptamers are single-stranded nucleic acid probes that can be evolved to have high specificity and affinity for different targets. These targets include biomar-ker proteins, small molecules, and even whole live cells that express a variety of surface proteins of interest. Aptamers offer several advantages over protein-based molecular probes such as low immunogenic activity, flexible modification, and in vitro synthesis. In addition, aptamers used as molecular probes can be made with easy signaling for binding with their corresponding targets. There are a few different fluorescence-based signal transduction mechanisms, such as direct fluorophore labeling, fluorescence resonance energy transfer (FRET), fluorescence quenching, fluorescence anisotropy, and light-switching excimers. These signaling processes in combination with various labeling strategies of nucleic acid aptamers contribute to simple, rapid, sensitive, and selective biological assays. In this chapter, we discuss the optical signaling of aptamers for single proteins such as ?-thrombin and platelet-derived growth factor (PDGF). We also present detailed discussion about fluorescent aptamers developed from cell-based systematic evolution of ligands by exponential enrichment (SELEX) for the recognition of different target tumor cells.

  11. Ultrathin oligonucleotide layers for fluorescence-based DNA sensors

    NASA Astrophysics Data System (ADS)

    Furch, M.; Ueberfeld, J.; Hartmann, Andreas; Bock, Daniel; Seeger, Stefan

    1996-11-01

    Preliminary investigations into the design of an affinity sensor using evanescent wave technology concentrate upon the means of immobilization of the receptor molecules. In this work DNA served as the selective recognition element. The molecular principle of a sequence-selective biosensor for DNA is based on a sandwich-hybridization assay wherein the analyte, a single-stranded (ss)DNA, bound specifically to both an immobilized capture probe and a dye-labeled oligonucleotide in free solution. The efficiency of the capture array depends on the density of highly organized oligonucleotides on the waveguide surface and correlates therefore directly with the specificity and the sensitivity of the sensor. In the present approach using the Langmuir- Blodgett technique cinnamoylbutylether-cellulose monolayers were transferred onto optical fibers or planar waveguides. These films served as matrices for the immobilization of biotinylated oligonucleotides via streptavidin. For the first time streptavidin was immobilized by that manner. The specificity of the streptavidin layer or the following bounded nucleic acid molecules were controlled by an enzyme- linked immunosorbent assay (ELISA). Finally, this application has also shown to be suitable for the detection of Salmonella, which is an important pathogen associated with acute gastroenteritidis and food borne diseases.

  12. Investigations of thermal conductivity of simple van der Waals crystal-based nanocomposites

    NASA Astrophysics Data System (ADS)

    Nikonkov, R. V.; Stachowiak, P.; Romanova, T. V.; Je?owski, A.; Sumarokov, V. V.

    2015-06-01

    The experimental setup for obtaining and determination of the thermal conductivity of simple van der Waals crystal-based nanocomposites is described. Preliminary thermal conductivity results of measurements carried out in the temperature range 1-40 K on two samples of methane crystals containing nanoparticles of hydroxyapatite are presented. These results confirm usability of the setup and its suitability as a proper experimental method for investigations of the thermal conductivity of the nanocomposites.

  13. Propeller design - a simple system based on model propeller test data III

    NASA Technical Reports Server (NTRS)

    Wieck, Fred E

    1926-01-01

    This report, the third of a series of four, describes a simple system for designing propellers of a standard form. In this report, the system is based on tests of a family of model propellers of standard Navy form, the data from which have been extended by means of calculations to cover the complete range likely to be found in practice. However, it can be worked out for any family having propellers of one general form.

  14. Simple BOTDA temperature sensor based on distributed Brillouin phase-shift measurements within a Sagnac interferometer

    NASA Astrophysics Data System (ADS)

    López-Gil, Alexia; Angulo-Vinuesa, Xabier; Dominguez-López, Alejandro; Martín-López, Sonia; González-Herráez, Miguel

    2015-09-01

    In this work we demonstrate an extremely simple BOTDA scheme capable of delivering distributed Brillouin Phase Shift measurements along an optical fiber. It is based on exploiting the non-reciprocity of the Stimulated Brillouin Scattering effect. This non-reciprocity is easily characterized by means of a suitably tuned Sagnac Interferometer. The technique is advantageous as, in comparison with previous methods, no complex modulation, no sharp filtering and no highbandwidth detection is needed. Theoretical and experimental proofs of the concept are given.

  15. A simple three-input DNA-based system works as a full-subtractor

    PubMed Central

    Lin, Hung-Yin; Chen, Jian-Zhou; Li, Hao-Yi; Yang, Chia-Ning

    2015-01-01

    Over the past decade, DNA has demonstrated remarkable potential in fabrication of molecular logic and arithmetic systems. In this work, a simple DNA-based system mimicking a full-subtractor that handles three inputs including one minuend and two subtrahends for eight input/output conditions is successfully designed. The whole system is established by one gate molecule and three input sequences, all made of single-stranded DNA sequences. PMID:26095534

  16. A simple modern correctness condition for a space-based high-performance multiprocessor

    NASA Technical Reports Server (NTRS)

    Probst, David K.; Li, Hon F.

    1992-01-01

    A number of U.S. national programs, including space-based detection of ballistic missile launches, envisage putting significant computing power into space. Given sufficient progress in low-power VLSI, multichip-module packaging and liquid-cooling technologies, we will see design of high-performance multiprocessors for individual satellites. In very high speed implementations, performance depends critically on tolerating large latencies in interprocessor communication; without latency tolerance, performance is limited by the vastly differing time scales in processor and data-memory modules, including interconnect times. The modern approach to tolerating remote-communication cost in scalable, shared-memory multiprocessors is to use a multithreaded architecture, and alter the semantics of shared memory slightly, at the price of forcing the programmer either to reason about program correctness in a relaxed consistency model or to agree to program in a constrained style. The literature on multiprocessor correctness conditions has become increasingly complex, and sometimes confusing, which may hinder its practical application. We propose a simple modern correctness condition for a high-performance, shared-memory multiprocessor; the correctness condition is based on a simple interface between the multiprocessor architecture and a high-performance, shared-memory multiprocessor; the correctness condition is based on a simple interface between the multiprocessor architecture and the parallel programming system.

  17. Winter wheat GPC estimation with fluorescence-based sensor measurements of canopy

    NASA Astrophysics Data System (ADS)

    Song, Xiaoyu; Wang, Jihua; Gu, Xiaohe; Xu, Xingang

    2015-10-01

    This study focused on the wheat grain protein content (GPC) estimation based on wheat canopy chlorophyll parameters which acquired by hand-held instrument, Multiplex 3. Nine fluorescence spectral indices from Multiplex sensor were used in this study. The wheat GPC estimation experiment was conducted in 2012 at the National Experiment Station for Precision Agriculture in Changping district, Beijing. A square with area of 1.1 ha was selected and divided to 110 small plots by 1010m in this study. In each plot, four 1-m2 area distributed in the square were selected for canopy fluorescence spectral measurements, physiological and biochemical analyses. Measurements were performed five times at wheat raising, jointing, heading stage, milking and ripening stage, respectively. The wheat plant samples for each plot were then collected after the measurement and sent to Lab for leaf N concentration (LNC) and canopy nitrogen density (CND) analyzed. GPC sampling for each plot was collected manually during the harvested season. Then, statistical analysis were performed to detect the correlation between fluorescence spectral indices and wheat CND for each growth stage, as well as GPC. The results indicate that two Nitrogen Balance Indices, NBI_G and NBI_R were more sensitive to wheat GPC than other fluorescence spectral indices at milking stage and ripening stage. Five linear regression models with GPC and fluorescence indices at different winter wheat growth stages were then established. The R2 of GPC estimated model increased form 0.312 at raising stage to 0.686 at ripening stage. The study reveals that canopy-level fluorescence spectral parameters were better indicators for the wheat group activity and could be demonstrated to be good indicators for winter wheat GPC estimation.

  18. Double-cladding-fiber-based detection system for intravascular mapping of fluorescent molecular probes

    NASA Astrophysics Data System (ADS)

    Razansky, R. Nika; Rozental, Amir; Mueller, Mathias S.; Deliolanis, Nikolaos; Jaffer, Farouc A.; Koch, Alexander W.; Ntziachristos, Vasilis

    2011-03-01

    Early detection of high-risk coronary atherosclerosis remains an unmet clinical challenge. We have previously demonstrated a near-infrared fluorescence catheter system for two-dimensional intravascular detection of fluorescence molecular probes [1]. In this work we improve the system performance by introducing a novel high resolution sensor. The main challenge of the intravascular sensor is to provide a highly focused spot at an application relevant distance on one hand and a highly efficient collection of emitted light on the other. We suggest employing a double cladding optical fiber (DCF) in combination with focusing optics to provide a sensor with both highly focused excitation light and highly efficient fluorescent light collection. The excitation laser is coupled into the single mode core of DCF and guided through a focusing element and a right angle prism. The resulting side-fired beam exhibits a small spot diameter (50 ?m) throughout a distance of up to 2 mm from the sensor. This is the distance of interest for intravascular coronary imaging application, determined by an average human coronary artery diameter. At the blood vessel wall, an activatable fluorescence molecular probe is excited in the diseased lesions. Next light of slightly shifted wavelength emits only in the places of the inflammations, associated with dangerous plaques [2]. The emitted light is collected by the cladding of the DCF, with a large collection angle (NA=0.4). The doublecladding acts as multimodal fiber and guides the collected light to the photo detection elements. The sensor automatically rotates and pulled-back, while each scanned point is mapped according to the amount of detected fluorescent emission. The resulting map of fluorescence activity helps to associate the atherosclerotic plaques with the inflammation process. The presented detection system is a valuable tool in the intravascular plaque detection and can help to differentiate the atherosclerotic plaques based on their biological activity, identify the ones that prone to rupture and therefore require more medical attention.

  19. Green fluorescent protein-based monitoring of endoplasmic reticulum redox poise

    PubMed Central

    Birk, Julia; Ramming, Thomas; Odermatt, Alex; Appenzeller-Herzog, Christian

    2013-01-01

    Pathological endoplasmic reticulum (ER) stress is tightly linked to the accumulation of reactive oxidants, which can be both upstream and downstream of ER stress. Accordingly, detrimental intracellular stress signals are amplified through establishment of a vicious cycle. An increasing number of human diseases are characterized by tissue atrophy in response to ER stress and oxidative injury. Experimental monitoring of stress-induced, time-resolved changes in ER reduction-oxidation (redox) states is therefore important. Organelle-specific examination of redox changes has been facilitated by the advent of genetically encoded, fluorescent probes, which can be targeted to different subcellular locations by means of specific amino acid extensions. These probes include redox-sensitive green fluorescent proteins (roGFPs) and the yellow fluorescent protein-based redox biosensor HyPer. In the case of roGFPs, variants with known specificity toward defined redox couples are now available. Here, we review the experimental framework to measure ER redox changes using ER-targeted fluorescent biosensors. Advantages and drawbacks of plate-reader and microscopy-based measurements are discussed, and the power of these techniques demonstrated in the context of selected cell culture models for ER stress. PMID:23781233

  20. Green fluorescent protein-based monitoring of endoplasmic reticulum redox poise.

    PubMed

    Birk, Julia; Ramming, Thomas; Odermatt, Alex; Appenzeller-Herzog, Christian

    2013-01-01

    Pathological endoplasmic reticulum (ER) stress is tightly linked to the accumulation of reactive oxidants, which can be both upstream and downstream of ER stress. Accordingly, detrimental intracellular stress signals are amplified through establishment of a vicious cycle. An increasing number of human diseases are characterized by tissue atrophy in response to ER stress and oxidative injury. Experimental monitoring of stress-induced, time-resolved changes in ER reduction-oxidation (redox) states is therefore important. Organelle-specific examination of redox changes has been facilitated by the advent of genetically encoded, fluorescent probes, which can be targeted to different subcellular locations by means of specific amino acid extensions. These probes include redox-sensitive green fluorescent proteins (roGFPs) and the yellow fluorescent protein-based redox biosensor HyPer. In the case of roGFPs, variants with known specificity toward defined redox couples are now available. Here, we review the experimental framework to measure ER redox changes using ER-targeted fluorescent biosensors. Advantages and drawbacks of plate-reader and microscopy-based measurements are discussed, and the power of these techniques demonstrated in the context of selected cell culture models for ER stress. PMID:23781233

  1. Nonlinear emission of quinolizinium-based dyes with application in fluorescence lifetime imaging.

    PubMed

    Marcelo, Gema; Pinto, Sandra; Caeque, Tatiana; Mariz, Ins F A; Cuadro, Ana M; Vaquero, Juan J; Martinho, Jos M G; Maas, Ermelinda M S

    2015-03-19

    Charged molecules based on the quinolizinum cation have potential applications as labels in fluorescence imaging in biological media under nonlinear excitation. A systematic study of the linear and nonlinear photophysics of derivatives of the quinolizinum cation substituted by either dimethylaniline or methoxyphenyl electron donors is performed. The effects of donor strength, conjugation length, and symmetry in the two-photon emission efficiency are analyzed in detail. The best performing nonlinear fluorophore, with two-photon absorption cross sections of 1140 GM and an emission quantum yield of 0.22, is characterized by a symmetric D-?-A(+)-?-D architecture based on the methoxyphenyl substituent. Application of this molecule as a fluorescent marker in optical microscopy of living cells revealed that, under favorable conditions, the fluorophore can be localized in the cytoplasmatic compartment of the cell, staining vesicular shape organelles. At higher dye concentrations and longer staining times, the fluorophore can also penetrate into the nucleus. The nonlinearly excited fluorescence lifetime imaging shows that the fluorophore lifetime is sensitive to its location in the different cell compartments. Using fluorescence lifetime microscopy, a multicolor map of the cell is drafted with a single dye. PMID:25135761

  2. Compact flashlamp-based fluorescence imager for use under ambient-light conditions

    NASA Astrophysics Data System (ADS)

    Lanni, Frederick; Pane, David A.; Weinstein, Shmuel J.; Waggoner, Alan S.

    2007-03-01

    A low-power, lightweight, multiwavelength fluorescence imager based on the use of a compact xenon flashlamp, bandpass filters, gated charge-coupled device camera, and digital image processing was developed for use on an autonomous rover vehicle. The imager discriminates against ambient light by use of microsecond excitation pulses along with synchronized camera operation to limit the time period in which ambient-light photocounts are accumulated, and digital image subtraction to remove background counts. In a 10 cm square field of view, weak fluorescence, equivalent to 0.05pmol fluorescein/mm2, can be quantified against a white-light background equivalent to shaded sunlight. For application in autonomous search for organisms in extreme environments such as in situ desert rock or soil, the instrument was equipped with a set of fluorescence excitation filters (380, 450, 545, and 600nm) and emission filters (460, 510, 620, and 740nm) suitable for detection of chlorophyll, applied stains for protein, DNA, lipid and carbohydrate, and autofluorescence. True-color images were obtained through red-green-blue imaging filters (630, 535, and 470nm) used with white-light flashes. Automated focusing on ground features was based on the R-band image and was carried out prior to fluorescence image acquisition.

  3. Compact flashlamp-based fluorescence imager for use under ambient-light conditions.

    PubMed

    Lanni, Frederick; Pane, David A; Weinstein, Shmuel J; Waggoner, Alan S

    2007-03-01

    A low-power, lightweight, multiwavelength fluorescence imager based on the use of a compact xenon flashlamp, bandpass filters, gated charge-coupled device camera, and digital image processing was developed for use on an autonomous rover vehicle. The imager discriminates against ambient light by use of microsecond excitation pulses along with synchronized camera operation to limit the time period in which ambient-light photocounts are accumulated, and digital image subtraction to remove background counts. In a 10 cm square field of view, weak fluorescence, equivalent to 0.05 pmol fluorescein/mm(2), can be quantified against a white-light background equivalent to shaded sunlight. For application in autonomous search for organisms in extreme environments such as in situ desert rock or soil, the instrument was equipped with a set of fluorescence excitation filters (380, 450, 545, and 600 nm) and emission filters (460, 510, 620, and 740 nm) suitable for detection of chlorophyll, applied stains for protein, DNA, lipid and carbohydrate, and autofluorescence. True-color images were obtained through red-green-blue imaging filters (630, 535, and 470 nm) used with white-light flashes. Automated focusing on ground features was based on the R-band image and was carried out prior to fluorescence image acquisition. PMID:17411186

  4. Image overlay solution based on threshold detection for a compact near infrared fluorescence goggle system

    PubMed Central

    Gao, Shengkui; Mondal, Suman B.; Zhu, Nan; Liang, RongGuang; Achilefu, Samuel; Gruev, Viktor

    2015-01-01

    Abstract. Near infrared (NIR) fluorescence imaging has shown great potential for various clinical procedures, including intraoperative image guidance. However, existing NIR fluorescence imaging systems either have a large footprint or are handheld, which limits their usage in intraoperative applications. We present a compact NIR fluorescence imaging system (NFIS) with an image overlay solution based on threshold detection, which can be easily integrated with a goggle display system for intraoperative guidance. The proposed NFIS achieves compactness, light weight, hands-free operation, high-precision superimposition, and a real-time frame rate. In addition, the miniature and ultra-lightweight light-emitting diode tracking pod is easy to incorporate with NIR fluorescence imaging. Based on experimental evaluation, the proposed NFIS solution has a lower detection limit of 25nM of indocyanine green at 27fps and realizes a highly precise image overlay of NIR and visible images of mice in vivo. The overlay error is limited within a 2-mm scale at a 65-cm working distance, which is highly reliable for clinical study and surgical use. PMID:25607724

  5. Fluorescence-based determination of the copper concentration in drinking water

    NASA Astrophysics Data System (ADS)

    Hötzer, Benjamin; Scheu, Timo; Jung, Gregor; Castritius, Stefan

    2013-05-01

    Copper is a heavy metal, which is used in heat and electrical conductors and in a multitude of alloys in the technical context. Moreover, it is a trace element that is essential for the life of organisms but can cause toxic effects in elevated concentrations. Maximum limits in water and beverages exist. Here, the decrease of the fluorescence lifetime of green fluorescent protein (GFP) by Förster resonance energy transfer is used to measure the copper ion concentration in drinking water. Therefore, a system is developed that is based on a GFP sample in a predefined concentration. The GFP mutant can be excited with blue light. For binding of copper ions, a His-tag is included in the GFP. After measuring the fluorescence lifetime of pure GFP, the copper determination of the sample is performed by lifetime measurement. Therefore, the lifetime can be assigned to the copper concentration of the GFP-doped drinking water sample. In summary, a method for the quantification of copper ions based on changes of the fluorescence lifetime of GFP is developed, and the measurement of the copper concentration in water samples is performed.

  6. Fluoromodule-based reporter/probes designed for in vivo fluorescence imaging.

    PubMed

    Zhang, Ming; Chakraborty, Subhasish K; Sampath, Padma; Rojas, Juan J; Hou, Weizhou; Saurabh, Saumya; Thorne, Steve H; Bruchez, Marcel P; Waggoner, Alan S

    2015-10-01

    Optical imaging of whole, living animals has proven to be a powerful tool in multiple areas of preclinical research and has allowed noninvasive monitoring of immune responses, tumor and pathogen growth, and treatment responses in longitudinal studies. However, fluorescence-based studies in animals are challenging because tissue absorbs and autofluoresces strongly in the visible light spectrum. These optical properties drive development and use of fluorescent labels that absorb and emit at longer wavelengths. Here, we present a far-red absorbing fluoromodule-based reporter/probe system and show that this system can be used for imaging in living mice. The probe we developed is a fluorogenic dye called SC1 that is dark in solution but highly fluorescent when bound to its cognate reporter, Mars1. The reporter/probe complex, or fluoromodule, produced peak emission near 730 nm. Mars1 was able to bind a variety of structurally similar probes that differ in color and membrane permeability. We demonstrated that a tool kit of multiple probes can be used to label extracellular and intracellular reporter-tagged receptor pools with 2 colors. Imaging studies may benefit from this far-red excited reporter/probe system, which features tight coupling between probe fluorescence and reporter binding and offers the option of using an expandable family of fluorogenic probes with a single reporter gene. PMID:26348895

  7. Fiber based in-vivo imaging of epithelial FAD fluorescence: experiments and simulations

    NASA Astrophysics Data System (ADS)

    Kanakaraj, Bala Nivetha; Narayanan Unni, Sujatha

    2015-03-01

    Fluorescence from endogenous fluorophores has been emerging as a promising biomarker for tissue discrimination resulting a noninvasive screening methodology to understand the biochemical and morphological variations in tissues associated with cancer development. We have developed a scan based fiber optic probe system to image increased flavin adenine dinucleotide (FAD) fluorescence from epithelial tissues under conditions mimicking dysplasia surrounded by normal tissues. Experiments were conducted on optical phantoms mimicking epithelial tissues excited by 450nm LED source. The spectral emission from the sample is collected via optical fibers and the imaging is performed by scanning the sample using a translation stage at desired resolution. Monte Carlo simulations were also performed by devising an optical model corresponding to epithelial tissue and the results were correlated with experimental fluorescence measurements. This whole field imaging approach could be useful for in vivo assessment of tissue pathologies based on auto fluorescence and can give a better quantitative approach for estimation of tissue properties by correlating the experimental and simulated data.

  8. Aptasensor for ampicillin using gold nanoparticle based dual fluorescence-colorimetric methods.

    PubMed

    Song, Kyung-Mi; Jeong, Euiyoung; Jeon, Weejeong; Cho, Minseon; Ban, Changill

    2012-02-01

    A gold nanoparticle based dual fluorescence-colorimetric method was developed as an aptasensor to detect ampicillin using its single-stranded DNA (ssDNA) aptamer, which was discovered by a magnetic bead-based SELEX technique. The selected aptamers, AMP4 (5'-CACGGCATGGTGGGCGTCGTG-3'), AMP17 (5'-GCGGGCGGTTGTATAGCGG-3'), and AMP18 (5'-TTAGTTGGGGTTCAGTTGG-3'), were confirmed to have high sensitivity and specificity to ampicillin (K(d), AMP7 = 9.4 nM, AMP17 = 13.4 nM, and AMP18 = 9.8 nM, respectively). The 5'-fluorescein amidite (FAM)-modified aptamer was used as a dual probe for observing fluorescence differences and color changes simultaneously. The lower limits of detection for this dual method were a 2 ng/mL by fluorescence and a 10 ng/mL by colorimetry for ampicillin in the milk as well as in distilled water. Because these detection limits were below the maximum residue limit of ampicillin, this aptasensor was sensitive enough to detect antibiotics in food products, such as milk and animal tissues. In addition, this dual aptasensor will be a more accurate method for antibiotics in food products as it concurrently uses two detection methods: fluorescence and colorimetry. PMID:22222912

  9. Fast-Response Calmodulin-Based Fluorescent Indicators Reveal Rapid Intracellular Calcium Dynamics

    PubMed Central

    Helassa, Nordine; Zhang, Xiao-hua; Conte, Ianina; Scaringi, John; Esposito, Elric; Bradley, Jonathan; Carter, Thomas; Ogden, David; Morad, Martin; Török, Katalin

    2015-01-01

    Faithful reporting of temporal patterns of intracellular Ca2+ dynamics requires the working range of indicators to match the signals. Current genetically encoded calmodulin-based fluorescent indicators are likely to distort fast Ca2+ signals by apparent saturation and integration due to their limiting fluorescence rise and decay kinetics. A series of probes was engineered with a range of Ca2+ affinities and accelerated kinetics by weakening the Ca2+-calmodulin-peptide interactions. At 37 °C, the GCaMP3-derived probe termed GCaMP3fast is 40-fold faster than GCaMP3 with Ca2+ decay and rise times, t1/2, of 3.3 ms and 0.9 ms, respectively, making it the fastest to-date. GCaMP3fast revealed discreet transients with significantly faster Ca2+ dynamics in neonatal cardiac myocytes than GCaMP6f. With 5-fold increased two-photon fluorescence cross-section for Ca2+ at 940 nm, GCaMP3fast is suitable for deep tissue studies. The green fluorescent protein serves as a reporter providing important novel insights into the kinetic mechanism of target recognition by calmodulin. Our strategy to match the probe to the signal by tuning the affinity and hence the Ca2+ kinetics of the indicator is applicable to the emerging new generations of calmodulin-based probes. PMID:26527405

  10. Fluoromodule-based reporter/probes designed for in vivo fluorescence imaging

    PubMed Central

    Zhang, Ming; Chakraborty, Subhasish K.; Sampath, Padma; Rojas, Juan J.; Hou, Weizhou; Saurabh, Saumya; Thorne, Steve H.; Bruchez, Marcel P.; Waggoner, Alan S.

    2015-01-01

    Optical imaging of whole, living animals has proven to be a powerful tool in multiple areas of preclinical research and has allowed noninvasive monitoring of immune responses, tumor and pathogen growth, and treatment responses in longitudinal studies. However, fluorescence-based studies in animals are challenging because tissue absorbs and autofluoresces strongly in the visible light spectrum. These optical properties drive development and use of fluorescent labels that absorb and emit at longer wavelengths. Here, we present a far-red absorbing fluoromodule–based reporter/probe system and show that this system can be used for imaging in living mice. The probe we developed is a fluorogenic dye called SC1 that is dark in solution but highly fluorescent when bound to its cognate reporter, Mars1. The reporter/probe complex, or fluoromodule, produced peak emission near 730 nm. Mars1 was able to bind a variety of structurally similar probes that differ in color and membrane permeability. We demonstrated that a tool kit of multiple probes can be used to label extracellular and intracellular reporter–tagged receptor pools with 2 colors. Imaging studies may benefit from this far-red excited reporter/probe system, which features tight coupling between probe fluorescence and reporter binding and offers the option of using an expandable family of fluorogenic probes with a single reporter gene. PMID:26348895

  11. Nicking enzyme-assisted biosensor for Salmonella enteritidis detection based on fluorescence resonance energy transfer.

    PubMed

    Song, Yang; Li, Wenkai; Duan, Yingfen; Li, Zhongjie; Deng, Le

    2014-05-15

    Salmonella enteritidis (S. enteritidis) outbreaks continue to occur, and have increased public awareness of this pathogen. Nicking endonuclease Nb.BbvC I is widely used for the detection of biomolecules and displays activity for specific double-stranded DNA (dsDNA). In this study, we developed a biosensor to detect S. enteritidis based on fluorescence resonance energy transfer (FRET) using nicking enzyme and carbon nanoparticles (CNPs). Because of the quenching effect of black hole quencher 1 (BHQ 1), the CNPs do not fluoresce in the reaction system. When the target bacteria are added, the nicking enzyme recognizes and cleaves the dsDNA fabricated by the interaction between probe and target. As a result, the CNPs dissociate from BHQ 1 and emit strong fluorescence. Using the nicking enzyme, the fluorescence signals of the biosensor are greatly amplified. The biosensor exhibited a linear relationship with the concentration of S. enteritidis ranging from 10(2) to 3 10(3)CFU/mL in water and from 1.5 10(2) to 3 10(3)CFU/mL in milk. The present results indicate that our FRET-based detection system can be widely employed for the effective detection of pathogens. PMID:24434495

  12. Image overlay solution based on threshold detection for a compact near infrared fluorescence goggle system

    NASA Astrophysics Data System (ADS)

    Gao, Shengkui; Mondal, Suman B.; Zhu, Nan; Liang, RongGuang; Achilefu, Samuel; Gruev, Viktor

    2015-01-01

    Near infrared (NIR) fluorescence imaging has shown great potential for various clinical procedures, including intraoperative image guidance. However, existing NIR fluorescence imaging systems either have a large footprint or are handheld, which limits their usage in intraoperative applications. We present a compact NIR fluorescence imaging system (NFIS) with an image overlay solution based on threshold detection, which can be easily integrated with a goggle display system for intraoperative guidance. The proposed NFIS achieves compactness, light weight, hands-free operation, high-precision superimposition, and a real-time frame rate. In addition, the miniature and ultra-lightweight light-emitting diode tracking pod is easy to incorporate with NIR fluorescence imaging. Based on experimental evaluation, the proposed NFIS solution has a lower detection limit of 25 nM of indocyanine green at 27 fps and realizes a highly precise image overlay of NIR and visible images of mice in vivo. The overlay error is limited within a 2-mm scale at a 65-cm working distance, which is highly reliable for clinical study and surgical use.

  13. A new fluorescent chemosensor for Al3+ ion based on schiff base naphthalene derivatives

    NASA Astrophysics Data System (ADS)

    Azadbakht, Reza; Rashidi, Somaye

    2014-06-01

    A new naphthalene derivative receptor (H2L) was synthesized. The chemosensor (H2L) exhibited a strong fluorescence enhancement in the presence of trace amounts of Al3+, attributable to chelation-enhanced fluorescence (CHEF) effect, which also displayed high selectivity over a series of other metal cations (Na+, K+, Cs+, Mg2+, Ba2+, Pb2+, Cr3+, Mn2+, Fe3+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+ and Ag+) in ethanol.

  14. Development of a competitive fluorescence-based synaptosome binding assay for brevetoxins

    PubMed Central

    McCall, Jennifer R.; Jacocks, Henry M.; Baden, Daniel G.; Bourdelais, Andrea J.

    2012-01-01

    Brevetoxins are a family of ladder-frame polyether toxins produced during blooms of the marine dinoflagellate Karenia brevis. Inhalation of brevetoxins aerosolized by wind and wave action can lead to asthma-like symptoms in beach goers. Consumption of either shellfish or finfish exposed to K. brevis blooms can lead to the development of neurotoxic shellfish poisoning. The toxic effects of brevetoxins are due to activation of voltage-sensitive sodium channels (VSSCs) in cell membranes. Binding of brevetoxin analogs and competitors to site 5 on these channels has historically been measured using a radioligand competition assay that is fraught with difficulty, including slow analysis time, production of radioactive waste, and cumbersome and expensive methods associated with the generation of radioactive labeled ligands. In this study, we describe the development of a novel fluorescent synaptosome binding assay for the brevetoxin receptor. BODIPY-conjugated to PbTx-2 was used as the labeled ligand. The BODIPY-PbTx-2 conjugate was found to displace [3H]-PbTx-3 from its binding site on VSSCs on rat brain synaptosomes with an equilibrium inhibition constant of 0.11 nM. We have shown that brevetoxin A and B analogs are all able to compete for binding with the fluorescent ligand. Most importantly, this assay was validated against the current site 5 receptor binding assay standard, the radioligand receptor assay for the brevetoxin receptor using [3H]-PbTx-3 as the labeled ligand. The fluorescence based assay yielded equilibrium inhibition constants comparable to the radioligand assay for all brevetoxin analogs. The fluorescence based assay was quicker, far less expensive, and did not generate radioactive waste or need radioactive facilities. As such, this fluorescence-based assay can be used to replace the current radioligand assay for site 5 on voltage-sensitive sodium channels and will be a vital tool for future experiments examining the binding affinity of various ligands for site 5 on sodium channels. PMID:22984362

  15. Development of a competitive fluorescence-based synaptosome binding assay for brevetoxins.

    PubMed

    McCall, Jennifer R; Jacocks, Henry M; Baden, Daniel G; Bourdelais, Andrea J

    2012-09-01

    Brevetoxins are a family of ladder-frame polyether toxins produced during blooms of the marine dinoflagellate Karenia brevis. Inhalation of brevetoxins aerosolized by wind and wave action can lead to asthma-like symptoms in beach goers. Consumption of either shellfish or finfish exposed to K. brevis blooms can lead to the development of neurotoxic shellfish poisoning. The toxic effects of brevetoxins are due to activation of voltage-sensitive sodium channels (VSSCs) in cell membranes. Binding of brevetoxin analogs and competitors to site 5 on these channels has historically been measured using a radioligand competition assay that is fraught with difficulty, including slow analysis time, production of radioactive waste, and cumbersome and expensive methods associated with the generation of radioactive labeled ligands. In this study, we describe the development of a novel fluorescent synaptosome binding assay for the brevetoxin receptor. BODIPY()-conjugated to PbTx-2 was used as the labeled ligand. The BODIPY()-PbTx-2 conjugate was found to displace [(3)H]-PbTx-3 from its binding site on VSSCs on rat brain synaptosomes with an equilibrium inhibition constant of 0.11 nM. We have shown that brevetoxin A and B analogs are all able to compete for binding with the fluorescent ligand. Most importantly, this assay was validated against the current site 5 receptor binding assay standard, the radioligand receptor assay for the brevetoxin receptor using [(3)H]-PbTx-3 as the labeled ligand. The fluorescence based assay yielded equilibrium inhibition constants comparable to the radioligand assay for all brevetoxin analogs. The fluorescence based assay was quicker, far less expensive, and did not generate radioactive waste or need radioactive facilities. As such, this fluorescence-based assay can be used to replace the current radioligand assay for site 5 on voltage-sensitive sodium channels and will be a vital tool for future experiments examining the binding affinity of various ligands for site 5 on sodium channels. PMID:22984362

  16. MATtrack: A MATLAB-Based Quantitative Image Analysis Platform for Investigating Real-Time Photo-Converted Fluorescent Signals in Live Cells.

    PubMed

    Courtney, Jane; Woods, Elena; Scholz, Dimitri; Hall, William W; Gautier, Virginie W

    2015-01-01

    We introduce here MATtrack, an open source MATLAB-based computational platform developed to process multi-Tiff files produced by a photo-conversion time lapse protocol for live cell fluorescent microscopy. MATtrack automatically performs a series of steps required for image processing, including extraction and import of numerical values from Multi-Tiff files, red/green image classification using gating parameters, noise filtering, background extraction, contrast stretching and temporal smoothing. MATtrack also integrates a series of algorithms for quantitative image analysis enabling the construction of mean and standard deviation images, clustering and classification of subcellular regions and injection point approximation. In addition, MATtrack features a simple user interface, which enables monitoring of Fluorescent Signal Intensity in multiple Regions of Interest, over time. The latter encapsulates a region growing method to automatically delineate the contours of Regions of Interest selected by the user, and performs background and regional Average Fluorescence Tracking, and automatic plotting. Finally, MATtrack computes convenient visualization and exploration tools including a migration map, which provides an overview of the protein intracellular trajectories and accumulation areas. In conclusion, MATtrack is an open source MATLAB-based software package tailored to facilitate the analysis and visualization of large data files derived from real-time live cell fluorescent microscopy using photoconvertible proteins. It is flexible, user friendly, compatible with Windows, Mac, and Linux, and a wide range of data acquisition software. MATtrack is freely available for download at eleceng.dit.ie/courtney/MATtrack.zip. PMID:26485569

  17. MATtrack: A MATLAB-Based Quantitative Image Analysis Platform for Investigating Real-Time Photo-Converted Fluorescent Signals in Live Cells

    PubMed Central

    Courtney, Jane; Woods, Elena; Scholz, Dimitri; Hall, William W.; Gautier, Virginie W.

    2015-01-01

    We introduce here MATtrack, an open source MATLAB-based computational platform developed to process multi-Tiff files produced by a photo-conversion time lapse protocol for live cell fluorescent microscopy. MATtrack automatically performs a series of steps required for image processing, including extraction and import of numerical values from Multi-Tiff files, red/green image classification using gating parameters, noise filtering, background extraction, contrast stretching and temporal smoothing. MATtrack also integrates a series of algorithms for quantitative image analysis enabling the construction of mean and standard deviation images, clustering and classification of subcellular regions and injection point approximation. In addition, MATtrack features a simple user interface, which enables monitoring of Fluorescent Signal Intensity in multiple Regions of Interest, over time. The latter encapsulates a region growing method to automatically delineate the contours of Regions of Interest selected by the user, and performs background and regional Average Fluorescence Tracking, and automatic plotting. Finally, MATtrack computes convenient visualization and exploration tools including a migration map, which provides an overview of the protein intracellular trajectories and accumulation areas. In conclusion, MATtrack is an open source MATLAB-based software package tailored to facilitate the analysis and visualization of large data files derived from real-time live cell fluorescent microscopy using photoconvertible proteins. It is flexible, user friendly, compatible with Windows, Mac, and Linux, and a wide range of data acquisition software. MATtrack is freely available for download at eleceng.dit.ie/courtney/MATtrack.zip. PMID:26485569

  18. On Development of a Problem Based Learning System for Linear Algebra with Simple Input Method

    NASA Astrophysics Data System (ADS)

    Yokota, Hisashi

    2011-08-01

    Learning how to express a matrix using a keyboard inputs requires a lot of time for most of college students. Therefore, for a problem based learning system for linear algebra to be accessible for college students, it is inevitable to develop a simple method for expressing matrices. Studying the two most widely used input methods for expressing matrices, a simpler input method for expressing matrices is obtained. Furthermore, using this input method and educator's knowledge structure as a concept map, a problem based learning system for linear algebra which is capable of assessing students' knowledge structure and skill is developed.

  19. Effects of communication and utility-based decision making in a simple model of evacuation

    NASA Astrophysics Data System (ADS)

    Smyrnakis, M.; Galla, T.

    2012-11-01

    We present a simple cellular automaton based model of decision making during evacuation. Evacuees have to choose between two different exit routes, resulting in a strategic decision making problem. Agents take their decisions based on utility functions, these can be revised as the evacuation proceeds, leading to complex interaction between individuals and to jamming transitions. The model also includes the possibility to communicate and exchange information with distant agents, information received may affect the decision of agents. We show that under a wider range of evacuation scenarios performance of the model system as a whole is optimal at an intermediate fraction of evacuees with access to communication.

  20. Fluorescence-based sensing of glucose using engineered glucose/galactose-binding protein: A comparison of fluorescence resonance energy transfer and environmentally sensitive dye labelling strategies

    SciTech Connect

    Khan, Faaizah; Gnudi, Luigi; Pickup, John C.

    2008-01-04

    Fluorescence-based glucose sensors using glucose-binding protein (GBP) as the receptor have employed fluorescence resonance energy transfer (FRET) and environmentally sensitive dyes, but with widely varying sensitivity. We therefore compared signal changes in (a) a FRET system constructed by transglutaminase-mediated N-terminal attachment of Alexa Fluor 488/555 as donor and QSY 7 as acceptor at Cys 152 or 182 mutations with (b) GBP labelled with the environmentally sensitive dye badan at C152 or 182. Both FRET systems had a small maximal fluorescence change at saturating glucose (7% and 16%), badan attached at C152 was associated with a 300% maximal fluorescence increase with glucose, though with badan at C182 there was no change. We conclude that glucose sensing based on GBP and FRET does not produce a larger enough signal change for clinical use; both the nature of the environmentally sensitive dye and its site of conjugation seem important for maximum signal change; badan-GBP152C has a large glucose-induced fluorescence change, suitable for development as a glucose sensor.