Modeling techniques and fluorescence imaging investigation of the interactions of an anthraquinone derivative with HSA and ctDNA

NASA Astrophysics Data System (ADS)

Fu, Zheng; Cui, Yanrui; Cui, Fengling; Zhang, Guisheng

2016-01-01

A new anthraquinone derivative (AORha) was synthesized. Its interactions with human serum albumin (HSA) and calf thymus DNA (ctDNA) were investigated by fluorescence spectroscopy, UV-visible absorption spectroscopy and molecular modeling. Cell viability assay and cell imaging experiment were performed using cervical cancer cells (HepG2 cells). The fluorescence results revealed that the quenching mechanism was static quenching. At different temperatures (290, 300, 310 K), the binding constants (K) and the number of binding sites (n) were determined, respectively. The positive ΔH and ΔS values showed that the binding of AORha with HSA was hydrophobic force, which was identical with the molecular docking result. Studying the fluorescence spectra, UV spectra and molecular modeling also verified that the binding mode of AORha and ctDNA might be intercalative. When HepG2 cells were treated with AORha, the fluorescence became brighter and turned green, which could be used for bioimaging.

Ex-vivo UV autofluorescence imaging and fluorescence spectroscopy of atherosclerotic pathology in human aorta

NASA Astrophysics Data System (ADS)

Lewis, William; Williams, Maura; Franco, Walfre

2017-02-01

The aim of our study was to identify fluorescence excitation-emission pairs correlated with atherosclerotic pathology in ex-vivo human aorta. Wide-field images of atherosclerotic human aorta were captured using UV and visible excitation and emission wavelength pairs of several known fluorophores to investigate correspondence with gross pathologic features. Fluorescence spectroscopy and histology were performed on 21 aortic samples. A matrix of Pearson correlation coefficients were determined for the relationship between relevant histologic features and the intensity of emission for 427 wavelength pairs. A multiple linear regression analysis indicated that elastin (370/460 nm) and tryptophan (290/340 nm) fluorescence predicted 58% of the variance in intima thickness (R-squared = 0.588, F(2,18) = 12.8, p=.0003), and 48% of the variance in media thickness (R-squared = 0.483, F(2,18) = 8.42, p=.002), suggesting that endogenous fluorescence intensity at these wavelengths can be utilized for improved pathologic characterization of atherosclerotic plaques.

Modeling techniques and fluorescence imaging investigation of the interactions of an anthraquinone derivative with HSA and ctDNA.

PubMed

Fu, Zheng; Cui, Yanrui; Cui, Fengling; Zhang, Guisheng

2016-01-15

A new anthraquinone derivative (AORha) was synthesized. Its interactions with human serum albumin (HSA) and calf thymus DNA (ctDNA) were investigated by fluorescence spectroscopy, UV-visible absorption spectroscopy and molecular modeling. Cell viability assay and cell imaging experiment were performed using cervical cancer cells (HepG2 cells). The fluorescence results revealed that the quenching mechanism was static quenching. At different temperatures (290, 300, 310 K), the binding constants (K) and the number of binding sites (n) were determined, respectively. The positive ΔH and ΔS values showed that the binding of AORha with HSA was hydrophobic force, which was identical with the molecular docking result. Studying the fluorescence spectra, UV spectra and molecular modeling also verified that the binding mode of AORha and ctDNA might be intercalative. When HepG2 cells were treated with AORha, the fluorescence became brighter and turned green, which could be used for bioimaging. Copyright © 2015 Elsevier B.V. All rights reserved.

Potential of ultraviolet wide-field imaging and multiphoton microscopy for analysis of dehydroergosterol in cellular membranes.

PubMed

Wüstner, Daniel; Brewer, Jonathan R; Bagatolli, Luis; Sage, Daniel

2011-01-01

Dehydroergosterol (DHE) is an intrinsically fluorescent sterol with absorption/emission in the ultraviolet (UV) region and biophysical properties similar to those of cholesterol. We compared the potential of UV-sensitive low-light-level wide-field (UV-WF) imaging with that of multiphoton (MP) excitation microscopy to monitor DHE in living cells. Significantly reduced photobleaching in MP microscopy of DHE enabled us to acquire three-dimensional z-stacks of DHE-stained cells and to obtain high-resolution maps of DHE in surface ruffles, nanotubes, and the apical membrane of epithelial cells. We found that the lateral resolution of MP microscopy is ∼1.5-fold higher than that of UV-WF deconvolution microscopy, allowing for improved spatiotemporal analysis of plasma membrane sterol distribution. Surface intensity patterns of DHE with a diameter of 0.2 μm persisting over several minutes could be resolved by MP time-lapse microscopy. Diffusion coefficients of 0.25-μm-diameter endocytic vesicles containing DHE were determined by MP spatiotemporal image correlation spectroscopy. The requirement of extremely high laser power for visualization of DHE by MP microscopy made this method less potent for multicolor applications with organelle markers like green fluorescent protein-tagged proteins. The signal-to-noise ratio obtainable by UV-WF imaging could be significantly improved by pixelwise bleach rate fitting and calculation of an amplitude image from the decay model and by frame averaging after pixelwise bleaching correction of the image stacks. We conclude that UV-WF imaging and MP microscopy of DHE provide complementary information regarding membrane distribution and intracellular targeting of sterols. © 2010 Wiley-Liss, Inc.

Vibrational and UV spectroscopic studies of 2-coumaranone by experimental and density functional theory calculations

NASA Astrophysics Data System (ADS)

Priya, Y. Sushma; Rao, K. Ramachandra; Chalapathi, P. V.; Satyavani, M.; Veeraiah, A.

2017-09-01

The vibrational and electronic properties of 2-coumaranone have been reported in the ground state using experimental techniques (FT-IR, FT-Raman, UV spectra and fluorescence microscopic imaging) and density functional theory (DFT) employing B3LYP correlation with the 6-31G(d, p) basis set. The theoretically reported optimized parameters, vibrational frequencies etc., were compared with the experimental values, which yielded good concurrence between the experimental and calculated values. The assignments of the vibrational spectra were done with the help of normal co-ordinate analysis (NCA) following the Scaled Quantum Mechanical Force Field(SQMFF) methodology. The whole assignments of fundamental modes were based on the potential energy distribution (PED) matrix. The electric dipole moment and the first order hyperpolarizability of the 2-coumaranone have been computed using quantum mechanical calculations. NBO and HOMO, LUMO analyses have been carried out. UV spectrum of 2-coumaranone was recorded in the region 100-300 nm and compared with the theoretical UV spectrum using TD-DFT and SAC-CI methods by which a good agreement is observed. Fluorescence microscopic imaging study reflects that the compound fluoresces in the green-yellow region.

Non-Euclidean phasor analysis for quantification of oxidative stress in ex vivo human skin exposed to sun filters using fluorescence lifetime imaging microscopy.

PubMed

Osseiran, Sam; Roider, Elisabeth M; Wang, Hequn; Suita, Yusuke; Murphy, Michael; Fisher, David E; Evans, Conor L

2017-12-01

Chemical sun filters are commonly used as active ingredients in sunscreens due to their efficient absorption of ultraviolet (UV) radiation. Yet, it is known that these compounds can photochemically react with UV light and generate reactive oxygen species and oxidative stress in vitro, though this has yet to be validated in vivo. One label-free approach to probe oxidative stress is to measure and compare the relative endogenous fluorescence generated by cellular coenzymes nicotinamide adenine dinucleotides and flavin adenine dinucleotides. However, chemical sun filters are fluorescent, with emissive properties that contaminate endogenous fluorescent signals. To accurately distinguish the source of fluorescence in ex vivo skin samples treated with chemical sun filters, fluorescence lifetime imaging microscopy data were processed on a pixel-by-pixel basis using a non-Euclidean separation algorithm based on Mahalanobis distance and validated on simulated data. Applying this method, ex vivo samples exhibited a small oxidative shift when exposed to sun filters alone, though this shift was much smaller than that imparted by UV irradiation. Given the need for investigative tools to further study the clinical impact of chemical sun filters in patients, the reported methodology may be applied to visualize chemical sun filters and measure oxidative stress in patients' skin. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Infrared images of reflection nebulae and Orion's bar: Fluorescent molecular hydrogen and the 3.3 micron feature

NASA Technical Reports Server (NTRS)

Burton, Michael G.; Moorhouse, Alan; Brand, P. W. J. L.; Roche, Patrick F.; Geballe, T. R.

1989-01-01

Images were obtained of the (fluorescent) molecular hydrogen 1-0 S(1) line, and of the 3.3 micron emission feature, in Orion's Bar and three reflection nebulae. The emission from these species appears to come from the same spatial locations in all sources observed. This suggests that the 3.3 micron feature is excited by the same energetic UV-photons which cause the molecular hydrogen to fluoresce.

Use of acousto-optic tunable filter in fluorescence imaging endoscopy

NASA Astrophysics Data System (ADS)

Bouhifd, Mounir; Whelan, Maurice; Aprahamian, Marc

2003-10-01

A prototype instrument for fluorescence-based medical diagnostics in vivo is described. The system consists of a rigid endoscope comprising a UV laser-source for fluorescence excitation and a white light source for direct imaging. An acousto-optic tuneable filter (AOTF) is employed as a full-field tuneable bandpass filter. This allows fast continuous or random-access tuning with high filtering efficiency. A study of the diagnostic potential of fluorescence imaging for pancreatitis was conducted on a rat model. In particular, the aim was to detect autofluorescence of endogenous protoporphyrin IX (PpIX) that has been shown to accumulate in early-stage diseased tissue undergoing an inflammatory response.

UV Raman and Fluorescence for Multi-Species Measurement in Hydrocarbon-Fueled High-Speed Propulsion

NASA Technical Reports Server (NTRS)

Skaggs, Patricia Annette; Nandula, Sastri P.; Pitz, Robert W.

1999-01-01

This report documents work performed through the NASA Graduate Student Researchers Program, Grant No. NGT3-52316. Research performed included investigation of two-line fluorescence imaging of OH for temperature measurement and an investigation of negative flame speeds for modeling of premixed turbulent flames. The laboratory work and initial analysis of the fluorescence imaging was performed at NASA Glen Research Center with follow up analysis at Vanderbilt University. The negative flame speed investigation was performed using an opposed jet flow simulation program at Vanderbilt University. The fluorescence imaging work is presented first followed by the negative flame speed investigation.

Deep UV Native Fluorescence Imaging of Antarctic Cryptoendolithic Communities

NASA Technical Reports Server (NTRS)

Storrie-Lombardi, M. C.; Douglas, S.; Sun, H.; McDonald, G. D.; Bhartia, R.; Nealson, K. H.; Hug, W. F.

2001-01-01

An interdisciplinary team at the Jet Propulsion Laboratory Center for Life Detection has embarked on a project to provide in situ chemical and morphological characterization of Antarctic cryptoendolithic microbial communities. We present here in situ deep ultraviolet (UV) native fluorescence and environmental scanning electron microscopy images transiting 8.5 mm into a sandstone sample from the Antarctic Dry Valleys. The deep ultraviolet imaging system employs 224.3, 248.6, and 325 nm lasers to elicit differential fluorescence and resonance Raman responses from biomolecules and minerals. The 224.3 and 248.6 nm lasers elicit a fluorescence response from the aromatic amino and nucleic acids. Excitation at 325 nm may elicit activity from a variety of biomolecules, but is more likely to elicit mineral fluorescence. The resultant fluorescence images provide in situ chemical and morphological maps of microorganisms and the associated organic matrix. Visible broadband reflectance images provide orientation against the mineral background. Environmental scanning electron micrographs provided detailed morphological information. The technique has made possible the construction of detailed fluorescent maps extending from the surface of an Antarctic sandstone sample to a depth of 8.5 mm. The images detect no evidence of microbial life in the superficial 0.2 mm crustal layer. The black lichen component between 0.3 and 0.5 mm deep absorbs all wavelengths of both laser and broadband illumination. Filamentous deep ultraviolet native fluorescent activity dominates in the white layer between 0.6 mm and 5.0 mm from the surface. These filamentous forms are fungi that continue into the red (iron-rich) region of the sample extending from 5.0 to 8.5 mm. Using differential image subtraction techniques it is possible to identify fungal nuclei. The ultraviolet response is markedly attenuated in this region, apparently from the absorption of ultraviolet light by iron-rich particles coating the filaments. Below 8.5 mm the filamentous morphology of the upper layers gives way to punctate 1-2 micron particles evidencing fluorescent activity following excitation at both deep ultraviolet wavelengths.

Characterization of E coli biofim formations on baby spinach leaf surfaces using hyperspectral fluorescence imaging

NASA Astrophysics Data System (ADS)

Cho, Hyunjeong; Baek, Insuck; Oh, Mirae; Kim, Sungyoun; Lee, Hoonsoo; Kim, Moon S.

2017-05-01

Bacterial biofilm formed by pathogens on fresh produce surfaces is a food safety concern because the complex extracellular matrix in the biofilm structure reduces the reduction and removal efficacies of washing and sanitizing processes such as chemical or irradiation treatments. Therefore, a rapid and nondestructive method to identify pathogenic biofilm on produce surfaces is needed to ensure safe consumption of fresh, raw produce. This research aimed to evaluate the feasibility of hyperspectral fluorescence imaging for detecting Escherichia.coli (ATCC 25922) biofilms on baby spinach leaf surfaces. Samples of baby spinach leaves were immersed and inoculated with five different levels (from 2.6x104 to 2.6x108 CFU/mL) of E.coli and stored at 4°C for 24 h and 48 h to induce biofilm formation. Following the two treatment days, individual leaves were gently washed to remove excess liquid inoculums from the leaf surfaces and imaged with a hyperspectral fluorescence imaging system equipped with UV-A (365 nm) and violet (405 nm) excitation sources to evaluate a spectral-image-based method for biofilm detection. The imaging results with the UV-A excitation showed that leaves even at early stages of biofilm formations could be differentiated from the control leaf surfaces. This preliminary investigation demonstrated the potential of fluorescence imaging techniques for detection of biofilms on leafy green surfaces.

Multimodal digital color imaging system for facial skin lesion analysis

NASA Astrophysics Data System (ADS)

Bae, Youngwoo; Lee, Youn-Heum; Jung, Byungjo

2008-02-01

In dermatology, various digital imaging modalities have been used as an important tool to quantitatively evaluate the treatment effect of skin lesions. Cross-polarization color image was used to evaluate skin chromophores (melanin and hemoglobin) information and parallel-polarization image to evaluate skin texture information. In addition, UV-A induced fluorescent image has been widely used to evaluate various skin conditions such as sebum, keratosis, sun damages, and vitiligo. In order to maximize the evaluation efficacy of various skin lesions, it is necessary to integrate various imaging modalities into an imaging system. In this study, we propose a multimodal digital color imaging system, which provides four different digital color images of standard color image, parallel and cross-polarization color image, and UV-A induced fluorescent color image. Herein, we describe the imaging system and present the examples of image analysis. By analyzing the color information and morphological features of facial skin lesions, we are able to comparably and simultaneously evaluate various skin lesions. In conclusion, we are sure that the multimodal color imaging system can be utilized as an important assistant tool in dermatology.

Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rastogi, Rajesh P.; Laboratory of Photobiology and Molecular Microbiology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005; Singh, Shailendra P.

2010-07-02

The generation of reactive oxygen species (ROS) under simulated solar radiation (UV-B: 0.30 Wm{sup -2}, UV-A: 25.70 Wm{sup -2} and PAR: 118.06 Wm{sup -2}) was studied in the cyanobacterium Anabaena variabilis PCC 7937 using the oxidant-sensing fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). DCFH-DA is a nonpolar dye, converted into the polar derivative DCFH by cellular esterases that are nonfluorescent but switched to highly fluorescent DCF when oxidized by intracellular ROS and other peroxides. The images obtained from the fluorescence microscope after 12 h of irradiation showed green fluorescence from cells covered with 295, 320 or 395 nm cut-off filters, indicating themore » generation of ROS in all treatments. However, the green/red fluorescence ratio obtained from fluorescence microscopic analysis showed the highest generation of ROS after UV-B radiation in comparison to PAR or UV-A radiation. Production of ROS was also measured by a spectrofluorophotometer and results obtained supported the results of fluorescence microscopy. Low levels of ROS were detected at the start (0 h) of the experiment showing that they are generated even during normal metabolism. This study also showed that UV-B radiation causes the fragmentation of the cyanobacterial filaments which could be due to the observed oxidative stress. This is the first report for the detection of intracellular ROS in a cyanobacterium by fluorescence microscopy using DCFH-DA and thereby suggesting the applicability of this method in the study of in vivo generation of ROS.« less

Applications of two-photon fluorescence microscopy in deep-tissue imaging

NASA Astrophysics Data System (ADS)

Dong, Chen-Yuan; Yu, Betty; Hsu, Lily L.; Kaplan, Peter D.; Blankschstein, D.; Langer, Robert; So, Peter T. C.

2000-07-01

Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.

CMOS image sensor-based implantable glucose sensor using glucose-responsive fluorescent hydrogel.

PubMed

Tokuda, Takashi; Takahashi, Masayuki; Uejima, Kazuhiro; Masuda, Keita; Kawamura, Toshikazu; Ohta, Yasumi; Motoyama, Mayumi; Noda, Toshihiko; Sasagawa, Kiyotaka; Okitsu, Teru; Takeuchi, Shoji; Ohta, Jun

2014-11-01

A CMOS image sensor-based implantable glucose sensor based on an optical-sensing scheme is proposed and experimentally verified. A glucose-responsive fluorescent hydrogel is used as the mediator in the measurement scheme. The wired implantable glucose sensor was realized by integrating a CMOS image sensor, hydrogel, UV light emitting diodes, and an optical filter on a flexible polyimide substrate. Feasibility of the glucose sensor was verified by both in vitro and in vivo experiments.

Synthesis of fluorescent carbon dots by a microwave heating process: structural characterization and cell imaging applications

NASA Astrophysics Data System (ADS)

Stefanakis, Dimitrios; Philippidis, Aggelos; Sygellou, Labrini; Filippidis, George; Ghanotakis, Demetrios; Anglos, Demetrios

2014-10-01

Two types of highly fluorescent carbon dots (C-dots) were prepared by a single-step procedure based on microwave heating citric acid and 6-aminocaproic acid or citric acid and urea in an aqueous solution. The small size of the isolated carbon dots along with their strong absorption in the UV and their excitation wavelength-dependent fluorescence render them ideal nanomaterials for biomedical applications (imaging and sensing). The structure and properties of the two types of C-dot materials were studied using a series of spectroscopic techniques. The ability of the C-dots to be internalized by HeLa cells was demonstrated via 3-photon fluorescence microscopy imaging.

Investigation of optical fibers for gas-phase, ultraviolet laser-induced-fluorescence (UV-LIF) spectroscopy.

PubMed

Hsu, Paul S; Kulatilaka, Waruna D; Jiang, Naibo; Gord, James R; Roy, Sukesh

2012-06-20

We investigate the feasibility of transmitting high-power, ultraviolet (UV) laser pulses through long optical fibers for laser-induced-fluorescence (LIF) spectroscopy of the hydroxyl radical (OH) and nitric oxide (NO) in reacting and non-reacting flows. The fundamental transmission characteristics of nanosecond (ns)-duration laser pulses are studied at wavelengths of 283 nm (OH excitation) and 226 nm (NO excitation) for state-of-the-art, commercial UV-grade fibers. It is verified experimentally that selected fibers are capable of transmitting sufficient UV pulse energy for single-laser-shot LIF measurements. The homogeneous output-beam profile resulting from propagation through a long multimode fiber is ideal for two-dimensional planar-LIF (PLIF) imaging. A fiber-coupled UV-LIF system employing a 6 m long launch fiber is developed for probing OH and NO. Single-laser-shot OH- and NO-PLIF images are obtained in a premixed flame and in a room-temperature NO-seeded N(2) jet, respectively. Effects on LIF excitation lineshapes resulting from delivering intense UV laser pulses through long fibers are also investigated. Proof-of-concept measurements demonstrated in the current work show significant promise for fiber-coupled UV-LIF spectroscopy in harsh diagnostic environments such as gas-turbine test beds.

5-ALA induced fluorescent image analysis of actinic keratosis

NASA Astrophysics Data System (ADS)

Cho, Yong-Jin; Bae, Youngwoo; Choi, Eung-Ho; Jung, Byungjo

2010-02-01

In this study, we quantitatively analyzed 5-ALA induced fluorescent images of actinic keratosis using digital fluorescent color and hyperspectral imaging modalities. UV-A was utilized to induce fluorescent images and actinic keratosis (AK) lesions were demarcated from surrounding the normal region with different methods. Eight subjects with AK lesion were participated in this study. In the hyperspectral imaging modality, spectral analysis method was utilized for hyperspectral cube image and AK lesions were demarcated from the normal region. Before image acquisition, we designated biopsy position for histopathology of AK lesion and surrounding normal region. Erythema index (E.I.) values on both regions were calculated from the spectral cube data. Image analysis of subjects resulted in two different groups: the first group with the higher fluorescence signal and E.I. on AK lesion than the normal region; the second group with lower fluorescence signal and without big difference in E.I. between two regions. In fluorescent color image analysis of facial AK, E.I. images were calculated on both normal and AK lesions and compared with the results of hyperspectral imaging modality. The results might indicate that the different intensity of fluorescence and E.I. among the subjects with AK might be interpreted as different phases of morphological and metabolic changes of AK lesions.

CMOS image sensor-based implantable glucose sensor using glucose-responsive fluorescent hydrogel

PubMed Central

Tokuda, Takashi; Takahashi, Masayuki; Uejima, Kazuhiro; Masuda, Keita; Kawamura, Toshikazu; Ohta, Yasumi; Motoyama, Mayumi; Noda, Toshihiko; Sasagawa, Kiyotaka; Okitsu, Teru; Takeuchi, Shoji; Ohta, Jun

2014-01-01

A CMOS image sensor-based implantable glucose sensor based on an optical-sensing scheme is proposed and experimentally verified. A glucose-responsive fluorescent hydrogel is used as the mediator in the measurement scheme. The wired implantable glucose sensor was realized by integrating a CMOS image sensor, hydrogel, UV light emitting diodes, and an optical filter on a flexible polyimide substrate. Feasibility of the glucose sensor was verified by both in vitro and in vivo experiments. PMID:25426316

Hyperspectral reflectance and fluorescence line-scan imaging system for online detection of fecal contamination on apples

NASA Astrophysics Data System (ADS)

Kim, Moon S.; Cho, Byoung-Kwan; Yang, Chun-Chieh; Chao, Kaunglin; Lefcourt, Alan M.; Chen, Yud-Ren

2006-10-01

We have developed nondestructive opto-electronic imaging techniques for rapid assessment of safety and wholesomeness of foods. A recently developed fast hyperspectral line-scan imaging system integrated with a commercial apple-sorting machine was evaluated for rapid detection of animal feces matter on apples. Apples obtained from a local orchard were artificially contaminated with cow feces. For the online trial, hyperspectral images with 60 spectral channels, reflectance in the visible to near infrared regions and fluorescence emissions with UV-A excitation, were acquired from apples moving at a processing sorting-line speed of three apples per second. Reflectance and fluorescence imaging required a passive light source, and each method used independent continuous wave (CW) light sources. In this paper, integration of the hyperspectral imaging system with the commercial applesorting machine and preliminary results for detection of fecal contamination on apples, mainly based on the fluorescence method, are presented.

Endogenous fluorescence emission of the ovary

NASA Astrophysics Data System (ADS)

Utzinger, Urs; Kirkpatrick, Nathaniel D.; Drezek, Rebekah A.; Brewer, Molly A.

2005-03-01

Epithelial ovarian cancer has the highest mortality rate among the gynecologic cancers. Early detection would significantly improve survival and quality of life of women at increased risk to develop ovarian cancer. We have constructed a device to investigate endogenous signals of the ovarian tissue surface in the UV C to visible range and describe our initial investigation of the use of optical spectroscopy to characterize the condition of the ovary. We have acquired data from more than 33 patients. A table top spectroscopy system was used to collect endogenous fluorescence with a fiberoptic probe that is compatible with endoscopic techniques. Samples were broken into five groups: Normal-Low Risk (for developing ovarian cancer) Normal-High Risk, Benign, and Cancer. Rigorous statistical analysis was applied to the data using variance tests for direct intensity versus diagnostic group comparisons and principal component analysis (PCA) to study the variance of the whole data set. We conclude that the diagnostically most useful excitation wavelengths are located in the UV. Furthermore, our results indicate that UV B and C are most useful. A safety analysis indicates that UV-C imaging can be conducted at exposure levels below safety thresholds. We found that fluorescence excited in the UV-C and UV-B range increases from benign to normal to cancerous tissues. This is in contrast to the emission created with UV-A excitation which decreased in the same order. We hypothesize that an increase of protein production and a decrease of fluorescence contributions of the extracellular matrix could explain this behavior. Variance analysis also identified fluctuation of fluorescence at 320/380 which is associated with collagen cross link residues. Small differences were observed between the group at high risk and normal risk for ovarian cancer. High risk samples deviated towards the cancer group and low risk samples towards benign group.

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.

Study of noninvasive detection of latent fingerprints using UV laser

NASA Astrophysics Data System (ADS)

Li, Hong-xia; Cao, Jing; Niu, Jie-qing; Huang, Yun-gang; Mao, Lin-jie; Chen, Jing-rong

2011-06-01

Latent fingerprints present a considerable challenge in forensics, and noninvasive procedure that captures a digital image of the latent fingerprints is significant in the field of criminal investigation. The capability of photography technologies using 266nm UV Nd:YAG solid state laser as excitation light source to provide detailed images of unprocessed latent fingerprints is demonstrated. Unprocessed latent fingerprints were developed on various non-absorbent and absorbing substrates. According to the special absorption, reflection, scattering and fluorescence characterization of the various residues in fingerprints (fatty acid ester, protein, and carbosylic acid salts etc) to the UV light to weaken or eliminate the background disturbance and increase the brightness contrast of fingerprints with the background, and using 266nm UV laser as excitation light source, fresh and old latent fingerprints on the surface of four types of non-absorbent objects as magazine cover, glass, back of cellphone, wood desktop paintwork and two types of absorbing objects as manila envelope, notebook paper were noninvasive detected and appeared through reflection photography and fluorescence photography technologies, and the results meet the fingerprint identification requirements in forensic science.

Facile synthesis of water-soluble and biocompatible fluorescent nitrogen-doped carbon dots for cell imaging.

PubMed

Wang, Weiping; Lu, Ya-Chun; Huang, Hong; Feng, Jiu-Ju; Chen, Jian-Rong; Wang, Ai-Jun

2014-04-07

A simple, facile and green hydrothermal method was developed in the synthesis of water-soluble nitrogen-doped carbon dots (N-CDs) from streptomycin. The as-prepared N-CDs displayed bright blue fluorescence under the irradiation of UV light, together with a high quantum yield of 7.6% and good biocompatibility as demonstrated by the cell viability assay. Thus, the N-CDs can be used as fluorescent probes for cell imaging, which have potential applications in bioimaging and related fields. This strategy opens a new way for the preparation of fluorescent carbon nanomaterials using small molecules as carbon sources.

The MIND PALACE: A Multi-Spectral Imaging and Spectroscopy Database for Planetary Science

NASA Astrophysics Data System (ADS)

Eshelman, E.; Doloboff, I.; Hara, E. K.; Uckert, K.; Sapers, H. M.; Abbey, W.; Beegle, L. W.; Bhartia, R.

2017-12-01

The Multi-Instrument Database (MIND) is the web-based home to a well-characterized set of analytical data collected by a suite of deep-UV fluorescence/Raman instruments built at the Jet Propulsion Laboratory (JPL). Samples derive from a growing body of planetary surface analogs, mineral and microbial standards, meteorites, spacecraft materials, and other astrobiologically relevant materials. In addition to deep-UV spectroscopy, datasets stored in MIND are obtained from a variety of analytical techniques obtained over multiple spatial and spectral scales including electron microscopy, optical microscopy, infrared spectroscopy, X-ray fluorescence, and direct fluorescence imaging. Multivariate statistical analysis techniques, primarily Principal Component Analysis (PCA), are used to guide interpretation of these large multi-analytical spectral datasets. Spatial co-referencing of integrated spectral/visual maps is performed using QGIS (geographic information system software). Georeferencing techniques transform individual instrument data maps into a layered co-registered data cube for analysis across spectral and spatial scales. The body of data in MIND is intended to serve as a permanent, reliable, and expanding database of deep-UV spectroscopy datasets generated by this unique suite of JPL-based instruments on samples of broad planetary science interest.

Fluorescence lifetime imaging and Fourier transform infrared spectroscopy of Michelangelo's David.

PubMed

Comelli, Daniela; Valentini, Gianluca; Cubeddu, Rinaldo; Toniolo, Lucia

2005-09-01

We developed a combined procedure for the analysis of works of art based on a portable system for fluorescence imaging integrated with analytical measurements on microsamples. The method allows us to localize and identify organic and inorganic compounds present on the surface of artworks. The fluorescence apparatus measures the temporal and spectral features of the fluorescence emission, excited by ultraviolet (UV) laser pulses. The kinetic of the emission is studied through a fluorescence lifetime imaging system, while an optical multichannel analyzer measures the fluorescence spectra of selected points. The chemical characterization of the compounds present on the artistic surfaces is then performed by means of analytical measurements on microsamples collected with the assistance of the fluorescence maps. The previous concepts have been successfully applied to study the contaminants on the surface of Michelangelo's David. The fluorescence analysis combined with Fourier transform infrared (FT-IR) measurements revealed the presence of beeswax, which permeates most of the statue surface, and calcium oxalate deposits mainly arranged in vertical patterns and related to rain washing.

VUV lithography

DOEpatents

George, Edward V.; Oster, Yale; Mundinger, David C.

1990-01-01

Deep UV projection lithography can be performed using an e-beam pumped solid excimer UV source, a mask, and a UV reduction camera. The UV source produces deep UV radiation in the range 1700-1300A using xenon, krypton or argon; shorter wavelengths of 850-650A can be obtained using neon or helium. A thin solid layer of the gas is formed on a cryogenically cooled plate and bombarded with an e-beam to cause fluorescence. The UV reduction camera utilizes multilayer mirrors having high reflectivity at the UV wavelength and images the mask onto a resist coated substrate at a preselected demagnification. The mask can be formed integrally with the source as an emitting mask.

Benzofurazan Sulfides for Thiol Imaging and Quantification in Live Cells through Fluorescence Microscopy

PubMed Central

Li, Yinghong; Yang, Yang; Guan, Xiangming

2012-01-01

Thiol groups play a significant role in various cellular functions. Cellular thiol concentrations can be affected by various physiological or pathological factors. A fluorescence imaging agent that can effectively and specifically image thiols in live cells through fluorescence microscopy is desirable for live cell thiol monitoring. Benzofurazan sulfides 1a–e were synthesized and found to be thiol specific fluorogenic agents except 1d. They are not fluorescent but form strong fluorescent thiol adducts after reacting with thiols through a sulfide-thiol exchange reaction. On the other hand, they exhibit no reaction with other biologically relevant nucleophilic functional groups such as -NH2, -OH, or -COOH revealing the specificity for the detection of thiols. Sulfide 1a was selected to confirm its ability to image cellular thiols through fluorescence microscopy. The compound was demonstrated to effectively image and quantify thiol changes in live cells through fluorescence microscopy using 430 nm and 520 nm as the excitation and emission wavelengths respectively. The quantification results of total thiol in live cells obtained from fluorescence microscopy were validated by an HPLC/UV total thiol assay method. The reagents and method will be of a great value to thiol redox-related research. PMID:22794193

Detection of melanomas by digital imaging of spectrally resolved UV light-induced autofluorescence of human skin

NASA Astrophysics Data System (ADS)

Chwirot, B. W.; Chwirot, S.; Jedrzejczyk, W.; Redzinski, J.; Raczynska, A. M.; Telega, K.

2001-07-01

We studied spectral and spatial distributions of the intensity of the ultraviolet light-excited fluorescence of human skin. Our studied performed in situ in 162 patients with malignant and non-malignant skin lesions resulted in a new method of detecting melanomas in situ using digital imaging of the spectrally resolved fluorescence. With our diagnostic algorithm we could successfully detect 88.5% of the cases of melanoma in the group of patients subject to examinations with the fluorescence method. A patent application for the method has been submitted to the Patent Office in Warsaw.

UV Raman imaging--a promising tool for astrobiology: comparative Raman studies with different excitation wavelengths on SNC Martian meteorites.

PubMed

Frosch, Torsten; Tarcea, Nicolae; Schmitt, Michael; Thiele, Hans; Langenhorst, Falko; Popp, Jürgen

2007-02-01

The great capabilities of UV Raman imaging have been demonstrated on the three Martian meteorites: Sayh al Uhaymir, Dar al Gani, and Zagami. Raman spectra without disturbing fluorescence and with high signal-to-noise-ratios and full of spectral features were derived. This result is of utmost importance for the development of powerful instruments for space missions. By point scanning the surfaces of the meteorite samples, it was possible for the first time to construct UV-Raman images out of the array of Raman spectra. Deep-UV Raman images are to the best of our knowledge presented for the first time. The images were used for a discussion of the chemical-mineralogical composition and texture of the meteorite surfaces. Comparative Raman studies applying visible and NIR Raman excitation wavelengths demonstrate a much better performance for UV Raman excitation. This comparative study of different Raman excitation wavelengths at the same sample spots was done by constructing a versatile, robust sample holder with a fixed micro-raster. The overall advantages of UV resonance Raman spectroscopy in terms of sensitivity and selectivity are demonstrated and discussed. Finally the application of this new technique for a UV Raman instrument for envisaged astrobiological focused space missions is suggested.

Multifunctional PHPMA-Derived Polymer for Ratiometric pH Sensing, Fluorescence Imaging, and Magnetic Resonance Imaging.

PubMed

Su, Fengyu; Agarwal, Shubhangi; Pan, Tingting; Qiao, Yuan; Zhang, Liqiang; Shi, Zhengwei; Kong, Xiangxing; Day, Kevin; Chen, Meiwan; Meldrum, Deirdre; Kodibagkar, Vikram D; Tian, Yanqing

2018-01-17

In this paper, we report synthesis and characterization of a novel multimodality (MRI/fluorescence) probe for pH sensing and imaging. A multifunctional polymer was derived from poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) and integrated with a naphthalimide-based-ratiometric fluorescence probe and a gadolinium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid complex (Gd-DOTA complex). The polymer was characterized using UV-vis absorption spectrophotometry, fluorescence spectrofluorophotometry, magnetic resonance imaging (MRI), and confocal microscopy for optical and MRI-based pH sensing and cellular imaging. In vitro labeling of macrophage J774 and esophageal CP-A cell lines shows the polymer's ability to be internalized in the cells. The transverse relaxation time (T 2 ) of the polymer was observed to be pH-dependent, whereas the spin-lattice relaxation time (T 1 ) was not. The pH probe in the polymer shows a strong fluorescence-based ratiometric pH response with emission window changes, exhibiting blue emission under acidic conditions and green emission under basic conditions, respectively. This study provides new materials with multimodalities for pH sensing and imaging.

VUV lithography

DOEpatents

George, E.V.; Oster, Y.; Mundinger, D.C.

1990-12-25

Deep UV projection lithography can be performed using an e-beam pumped solid excimer UV source, a mask, and a UV reduction camera. The UV source produces deep UV radiation in the range 1,700--1,300A using xenon, krypton or argon; shorter wavelengths of 850--650A can be obtained using neon or helium. A thin solid layer of the gas is formed on a cryogenically cooled plate and bombarded with an e-beam to cause fluorescence. The UV reduction camera utilizes multilayer mirrors having high reflectivity at the UV wavelength and images the mask onto a resist coated substrate at a preselected demagnification. The mask can be formed integrally with the source as an emitting mask. 6 figs.

Investigation of optical fibers for high-repetition-rate, ultraviolet planar laser-induced fluorescence of OH.

PubMed

Hsu, Paul S; Kulatilaka, Waruna D; Roy, Sukesh; Gord, James R

2013-05-01

We investigate the fundamental transmission characteristics of nanosecond-duration, 10 kHz repetition rate, ultraviolet (UV) laser pulses through state-of-the-art, UV-grade fused-silica fibers being used for hydroxyl radical (OH) planar laser-induced fluorescence (PLIF) imaging. Studied in particular are laser-induced damage thresholds (LIDTs), nonlinear absorption, and optical transmission stability during long-term UV irradiation. Solarization (photodegradation) effects are significantly enhanced when the fiber is exposed to high-repetition-rate, 283 nm UV irradiation. For 10 kHz laser pulses, two-photon absorption is strong and LIDTs are low, as compared to those of laser pulses propagating at 10 Hz. The fiber characterization results are utilized to perform single-laser-shot, OH-PLIF imaging in pulsating turbulent flames with a laser that operates at 10 kHz. The nearly spatially uniform output beam that exits a long multimode fiber becomes ideal for PLIF measurements. The proof-of-concept measurements show significant promise for extending the application of a fiber-coupled, high-speed OH-PLIF system to harsh environments such as combustor test beds, and potential system improvements are suggested.

Facile synthesis of Curcuma longa tuber powder engineered metal nanoparticles for bioimaging applications

NASA Astrophysics Data System (ADS)

Sankar, Renu; Rahman, Pattanathu K. S. M.; Varunkumar, Krishnamoorthy; Anusha, Chidambaram; Kalaiarasi, Arunachalam; Shivashangari, Kanchi Subramanian; Ravikumar, Vilwanathan

2017-02-01

Nanomaterials based fluorescent agents are rapidly becoming significant and promising transformative tools for improving medical diagnostics for extensive in vivo imaging modalities. Compared with conventional fluorescent agents, nano-fluorescence has capabilities to improve the in vivo detection and enriched targeting efficiencies. In our laboratory we synthesized fluorescent metal nanoparticles of silver, copper and iron using Curcuma longa tuber powder by simple reduction. The physicochemical properties of the synthesized metal nanoparticles were attained using UV-visible spectrophotometry, scanning electron microscopy with EDAX spectroscopy, dynamic light scattering, Fourier-transform infrared spectroscopy and X-ray diffraction. The Curcuma longa tuber powder has one of the bioactive compound Curcumin might act as a capping agent during the synthesis of nanoparticles. The synthesized metal nanoparticles fluorescence property was confirmed by spectrofluorometry. When compared with copper and iron nanoparticles the silver nanoparticles showed high fluorescence intensity under spectrofluorometry. Moreover, in vitro cell images of the silver nanoparticles in A549 cell lines also correlated with the results of spectrofluorometry. These silver nanoparticles show inspiring cell-imaging applications. They enter into cells without any further modifications, and the fluorescence property can be utilized for fluorescence-based cell imaging applications.

Tuning of gold nanoclusters sensing applications with bovine serum albumin and bromelain for detection of Hg2+ ion and lambda-cyhalothrin via fluorescence turn-off and on mechanisms.

PubMed

Bhamore, Jigna R; Jha, Sanjay; Basu, Hirakendu; Singhal, Rakesh Kumar; Murthy, Z V P; Kailasa, Suresh Kumar

2018-04-01

Herein, fluorescent gold nanoclusters (Au NCs) were obtained by one-pot synthetic method using bovine serum albumin (BSA) and bromelain as templates. As-synthesized fluorescent Au NCs were stable and showed bright red fluorescence under UV lamp at 365 nm. The fluorescent Au NCs exhibit the emission intensity at 648 nm when excited at 498 nm. Various techniques were used such as spectroscopy (UV-visible, fluorescence, and Fourier-transform infrared), high-resolution transmission electron microscopy, and dynamic light scattering for the characterization of fluorescent Au NCs. The values of I 0 /I at 648 nm are proportional to the concentrations of Hg 2+ ion in the range from 0.00075 to 5.0 μM and of lambda-cyhalothrin in the range from 0.01 to 10 μM with detection limits of 0.0003 and 0.0075 μM for Hg 2+ ion and lambda-cyhalothrin, respectively. The practical application of the probe was successfully demonstrated by analyzing Hg 2+ ion and lambda-cyhalothrin in water samples. In addition, Au NCs used as probes for imaging of Simplicillium fungal cells. These results indicated that the as-synthesized Au NCs have proven to be promising fluorescent material for the sensing of Hg 2+ ion and lambda-cyhalothrin in environmental and for imaging of microorganism cells in biomedical applications.

Fluorescent Microscopy Enhancement Using Imaging

NASA Astrophysics Data System (ADS)

Conrad, Morgan P.; Reck tenwald, Diether J.; Woodhouse, Bryan S.

1986-06-01

To enhance our capabilities for observing fluorescent stains in biological systems, we are developing a low cost imaging system based around an IBM AT microcomputer and a commercial image capture board compatible with a standard RS-170 format video camera. The image is digitized in real time with 256 grey levels, while being displayed and also stored in memory. The software allows for interactive processing of the data, such as histogram equalization or pseudocolor enhancement of the display. The entire image, or a quadrant thereof, can be averaged over time to improve the signal to noise ratio. Images may be stored to disk for later use or comparison. The camera may be selected for better response in the UV or near IR. Combined with signal averaging, this increases the sensitivity relative to that of the human eye, while still allowing for the fluorescence distribution on either the surface or internal cytoskeletal structure to be observed.

Selective visualization of fluorescent sterols in Caenorhabditis elegans by bleach-rate-based image segmentation.

PubMed

Wüstner, Daniel; Landt Larsen, Ane; Faergeman, Nils J; Brewer, Jonathan R; Sage, Daniel

2010-04-01

The nematode Caenorhabditis elegans is a genetically tractable model organism to investigate sterol transport. In vivo imaging of the fluorescent sterol, dehydroergosterol (DHE), is challenged by C. elegans' high autofluorescence in the same spectral region as emission of DHE. We present a method to detect DHE selectively, based on its rapid bleaching kinetics compared to cellular autofluorescence. Worms were repeatedly imaged on an ultraviolet-sensitive wide field (UV-WF) microscope, and bleaching kinetics of DHE were fitted on a pixel-basis to mathematical models describing the intensity decay. Bleach-rate constants were determined for DHE in vivo and confirmed in model membranes. Using this method, we could detect enrichment of DHE in specific tissues like the nerve ring, the spermateca and oocytes. We confirm these results in C. elegans gut-granule-loss (glo) mutants with reduced autofluorescence and compare our method with three-photon excitation microscopy of sterol in selected tissues. Bleach-rate-based UV-WF imaging is a useful tool for genetic screening experiments on sterol transport, as exemplified by RNA interference against the rme-2 gene coding for the yolk receptor and for worm homologues of Niemann-Pick C disease proteins. Our approach is generally useful for identifying fluorescent probes in the presence of high cellular autofluorescence.

Synthesis of Water Dispersible Fluorescent Carbon Nanocrystals from Syzygium cumini Fruits for the Detection of Fe3+ Ion in Water and Biological Samples and Imaging of Fusarium avenaceum Cells.

PubMed

Bhamore, Jigna R; Jha, Sanjay; Singhal, Rakesh Kumar; Kailasa, Suresh Kumar

2017-01-01

In this work, water dispersible fluorescent carbon nanocrystals (NCs) were synthesized by a simple, green and low cost hydrothermal method using Syzygium cumini (jamun) as a carbon source at 180 °C for 6 h. The average size of carbon NCs was found to be 2.1 ± 0.5 nm and shown bright blue fluorescence when excited at 365 nm under UV lamp. The carbon NCs were characterized by spectroscopic (UV-visible and fluorescence, Fourier transform infrared and dynamic light scattering) and high resolution transmission electron microscopic techniques. The quantum yield of carbon NCs was found to be ~5.9 % at 438 nm emission wavelength when excited at 360 nm. It was noticed that none of the metal ions quenched the fluorescence intensity of carbon NCs at 438 nm except for Fe 3+ , indicating the formation of Fe 3+ ion-carbon NCs complexes. The linear range was observed in the concentration range of 0.01-100 μM with the corresponding detection limits of 0.001 μM, respectively. Furthermore, the carbon NCs were used as probes for imaging of fungal (Fusarium avenaceum) cells.

UV reflection properties of plumage and skin of domesticated turkeys (Meleagris gallopavo f. dom.) as revealed by UV photography.

PubMed

Bartels, T; Lütgeharm, J-H; Wähner, M; Berk, J

2017-12-01

Reflection and fluorescence properties of feathered and non-feathered body regions of white- and bronze-colored fattening turkeys of various ages were examined by ultraviolet (UV) photography. The examinations were carried out on 20 white-feathered fattening turkeys (B.U.T. 6; 10 males, 10 females) and 20 bronze-feathered fattening turkeys (Grelier 708; 10 males, 10 females) over a period of 21 weeks. The turkeys were photographed once a wk under long-wave UV (UVA) radiation illumination (λ = 344-407 nm) using a digital camera. A bandpass filter was used for UV reflectography to filter out the visible components of the used light source. A longpass filter was used for UV fluorescence photography to avoid blurring in the image due to chromatic aberration as a result of UV illumination. We found that natal down feathers of white-feathered turkeys showed an intense yellowish-green fluorescence under UVA light. UVA fluorescence also was shown by the natal downs of the slightly melanized plumage areas of bronze turkeys. Vaned feathers of white fattening turkeys reflected UVA radiation. Freshly molted feathers were optically distinguishable from the previous feather generation due to their more intense UVA reflection. In bronze turkeys, both the bright end seams of the dark pennaceous feathers and rectrices and the bright banding of primary and secondary remiges reflected UVA radiation. Intense UVA fluorescence was recognizable in day-old chicks of both color variants on the scutellate scales of the legs and toes. In male turkeys of both color variants, UVA-reflecting parts were recognizable with increasing age on the featherless head region. The UVA-fluorescent and UVA-reflective characteristics of the plumage of fattening turkeys were closely related to the plumage color, the feather type, the molting state, and the age of the birds. Further research is needed regarding the UVA-reflecting properties of the turkey plumage and the effects of full-spectrum illumination, including the UVA spectrum, on the behavior and health of fattening turkeys. © 2017 Poultry Science Association Inc.

A Novel Water-Soluble Fluorescence Probe with Wash-Free Cellular Imaging Capacity Based on AIE Characteristics.

PubMed

Qian, Yunxia; Liu, Hongmei; Tan, Haijian; Yang, Qingmin; Zhang, Shuchen; Han, Lingui; Yi, Xuegang; Huo, Li; Zhao, Hongchi; Wu, Yonggang; Bai, Libin; Ba, Xinwu

2017-05-01

A potential real-time imaging water-soluble fluorescent polymer (P3) is facilely prepared via one-pot method. For P3, tetraphenylethene unit serves as the fluorescent unit, poly(acryloyl ethylene diamine) (a kind of polyelectrolyte) with specific degree of polymerization acts as water-soluble part. 1 H-NMR, gel permeation chromatography (GPC), UV-vis spectroscopy, photoluminescence (PL), and confocal laser scanning microscopy are undertaken to characterize the structure and property of P3. The results of wash-free cellular imaging show that the signal-to-noise ratio is high as the concentration of P3 is 50 μg mL -1 . In addition, the pH-responsive and Cd 2+ -responsive are also investigated in this paper. The results coming from pH-responsive show that P3 solution displays significant fluorescence under near neutral. And the result from the cellular imaging shows that intracellular fluorescence intensity enhances with the augment of concentration of Cd 2+ , which reveals that P3 can give a hint to resolve the dilemma of traditional fluorescent dyes used as living cellular fluorescent probe. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Corneal epithelium and UV-protection of the eye.

PubMed

Ringvold, A

1998-04-01

To study UV-absorption and UV-induced fluorescence in the bovine corneal epithelium. Spectrophotometry and spectrofluorimetry. The corneal epithelium absorbs UV-B radiation mainly owing to its content of protein, RNA, and ascorbate. Some of the absorbed energy is transformed to the less biotoxic UV-A radiation by fluorescence. RNA and ascorbate reduce tissue fluorescence. The corneal epithelium acts as a UV-filter, protecting internal eye structures through three different mechanisms: (1) Absorption of UV-B roughly below 310 nm wavelength. (2) Fluorescence-mediated ray transformation to longer wavelengths. (3) Fluorescence reduction. The extremely high ascorbate concentration in the corneal epithelium has a key role in two of these processes.

The total antioxidant capacity and fluorescence imaging of selected plant leaves commonly consumed in Brunei Darussalam

NASA Astrophysics Data System (ADS)

Watu, Aswani; Metussin, Nurzaidah; Yasin, Hartini M.; Usman, Anwar

2018-02-01

We investigated the total antioxidant capacity and fluorescence imaging of several selected plants, namely Centella asiatica, Aidia borneensis and Anacardium occidentale, which are grown and traditionally consumed in Brunei Darussalam. The total antioxidant capacities of aqueous-methanolic infusions of their leaves were measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and microscopic fluorescence images were measured to identify the fluorescent substances bound in the leaves. We found that the total antioxidant capacity of their infusions is estimated to be 150, 25, 15 folds, respectively, lower compared with that of the standard gallic acid. Accordingly, we demonstrated that the relative antioxidant activity of young and matured leaves agrees with the intensity of red light emission of their fresh leaves upon UV excitation. Thus, this non-invasive spectroscopic method can be potentially utilized to indicate the antioxidants in plant leaves qualitatively.

One-Step Synthesis of Fluorescent Boron Nitride Quantum Dots via a Hydrothermal Strategy Using Melamine as Nitrogen Source for the Detection of Ferric Ions.

PubMed

Huo, Bingbing; Liu, Bingping; Chen, Tao; Cui, Liang; Xu, Gengfang; Liu, Mengli; Liu, Jingquan

2017-10-10

A facile and effective approach for the preparation of functionalized born nitride quantum dots (BNQDs) with blue fluorescence was explored by the hydrothermal treatment of the mixture of boric acid and melamine at 200 °C for 15 h. The as-prepared BNQDs were characterized by transmission electron microscopy (TEM), high-resolution TEM, atomic force microscopy, X-ray photoelectron spectroscopy, UV-vis spectroscopy, and fluorescence spectroscopy. The single layered BNQDs with the average size of 3 nm showed a blue light emission under the illumination of the UV light. The BNQDs could be easily dispersed in an aqueous medium and applied as fluorescent probes for selective detection of Fe 3+ with remarkable selectivity and sensitivity (the lowest detection limit was 0.3 μM). The fluorescence fiber imaging demonstrated that the as-prepared quantum dots could be used as a valuable fluorchrome. Therefore, the BNQDs could be envisioned for potential applications in many fields such as biocompatible staining, fluorescent probes, and biological labeling.

Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.

PubMed

Diaspro, A; Corosu, M; Ramoino, P; Robello, M

1999-11-01

Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) pumped by a high-power (5 W, 532 nm) laser (Millennia V, Spectra Physics Inc.) and an ultracompact confocal scanning head, Nikon PCM2000 (Nikon Instruments, Florence, Italy) using a single-pinhole design. Three-dimensional point-spread function has been measured to define spatial resolution performances. The TPE microscope has been used with a wide range of excitable fluorescent molecules (DAPI, Fura-2, Indo-1, DiOC(6)(3), fluoresceine, Texas red) covering a single photon spectral range from UV to green. An example is reported on 3D imaging of the helical structure of the sperm head of the Octopus Eledone cirrhosa labelled with an UV excitable dye, i.e., DAPI. The system can be easily switched for operating both in conventional and two-photon mode. Copyright 1999 Wiley-Liss, Inc.

A randomized control trial evaluating fluorescent ink versus dark ink tattoos for breast radiotherapy

PubMed Central

Kirby, Anna M; Lee, Steven F; Bartlett, Freddie; Titmarsh, Kumud; Donovan, Ellen; Griffin, Clare L; Gothard, Lone; Locke, Imogen; McNair, Helen A

2016-01-01

Objective: The purpose of this UK study was to evaluate interfraction reproducibility and body image score when using ultraviolet (UV) tattoos (not visible in ambient lighting) for external references during breast/chest wall radiotherapy and compare with conventional dark ink. Methods: In this non-blinded, single-centre, parallel group, randomized control trial, patients were allocated to receive either conventional dark ink or UV ink tattoos using computer-generated random blocks. Participant assignment was not masked. Systematic (∑) and random (σ) setup errors were determined using electronic portal images. Body image questionnaires were completed at pre-treatment, 1 month and 6 months to determine the impact of tattoo type on body image. The primary end point was to determine that UV tattoo random error (σsetup) was no less accurate than with conventional dark ink tattoos, i.e. <2.8 mm. Results: 46 patients were randomized to receive conventional dark or UV ink tattoos. 45 patients completed treatment (UV: n = 23, dark: n = 22). σsetup for the UV tattoo group was <2.8 mm in the u and v directions (p = 0.001 and p = 0.009, respectively). A larger proportion of patients reported improvement in body image score in the UV tattoo group compared with the dark ink group at 1 month [56% (13/23) vs 14% (3/22), respectively] and 6 months [52% (11/21) vs 38% (8/21), respectively]. Conclusion: UV tattoos were associated with interfraction setup reproducibility comparable with conventional dark ink. Patients reported a more favourable change in body image score up to 6 months following treatment. Advances in knowledge: This study is the first to evaluate UV tattoo external references in a randomized control trial. PMID:27710100

A randomized control trial evaluating fluorescent ink versus dark ink tattoos for breast radiotherapy.

PubMed

Landeg, Steven J; Kirby, Anna M; Lee, Steven F; Bartlett, Freddie; Titmarsh, Kumud; Donovan, Ellen; Griffin, Clare L; Gothard, Lone; Locke, Imogen; McNair, Helen A

2016-12-01

The purpose of this UK study was to evaluate interfraction reproducibility and body image score when using ultraviolet (UV) tattoos (not visible in ambient lighting) for external references during breast/chest wall radiotherapy and compare with conventional dark ink. In this non-blinded, single-centre, parallel group, randomized control trial, patients were allocated to receive either conventional dark ink or UV ink tattoos using computer-generated random blocks. Participant assignment was not masked. Systematic (∑) and random (σ) setup errors were determined using electronic portal images. Body image questionnaires were completed at pre-treatment, 1 month and 6 months to determine the impact of tattoo type on body image. The primary end point was to determine that UV tattoo random error (σ setup ) was no less accurate than with conventional dark ink tattoos, i.e. <2.8 mm. 46 patients were randomized to receive conventional dark or UV ink tattoos. 45 patients completed treatment (UV: n = 23, dark: n = 22). σ setup for the UV tattoo group was <2.8 mm in the u and v directions (p = 0.001 and p = 0.009, respectively). A larger proportion of patients reported improvement in body image score in the UV tattoo group compared with the dark ink group at 1 month [56% (13/23) vs 14% (3/22), respectively] and 6 months [52% (11/21) vs 38% (8/21), respectively]. UV tattoos were associated with interfraction setup reproducibility comparable with conventional dark ink. Patients reported a more favourable change in body image score up to 6 months following treatment. Advances in knowledge: This study is the first to evaluate UV tattoo external references in a randomized control trial.

The preparation of ethylenediamine-modified fluorescent carbon dots and their use in imaging of cells.

PubMed

Dong, Wei; Zhou, Siqi; Dong, Yan; Wang, Jingwen; Ge, Xin; Sui, Lili

2015-09-01

In this work, fluorescent carbon dots (CDs) were synthesized using a hydrothermal method with glucose as the carbon source and were surface-modified with ethylenediamine. The properties of as-prepared CDs were analyzed by transmission electron microscopy (TEM), Fourier transform infrared (FTIR), ultraviolet-visible light (UV/vis) absorption and fluorescent spectra. Furthermore, CDs conjugated with mouse anti-(human carcinoembryonic antigen) (CEA) monoclonal antibody were successful employed in the biolabeling and fluorescent imaging of human gastric carcinoma cells. In addition, the cytotoxicity of CDs was also tested using human gastric carcinoma cells. There was no apparent cytotoxicity on human gastric carcinoma cells. These results suggest the potential application of the as-prepared CDs in bioimaging and related fields. Copyright © 2015 John Wiley & Sons, Ltd.

Studies on Structural, Optical, Thermal and Electrical Properties of Perylene-Doped p-terphenyl Luminophors.

PubMed

Desai, Netaji K; Mahajan, Prasad G; Bhopate, Dhanaji P; Dalavi, Dattatray K; Kamble, Avinash A; Gore, Anil H; Dongale, Tukaram D; Kolekar, Govind B; Patil, Shivajirao R

2018-01-01

A simple solid state reaction technique was employed for the preparation of polycrystalline luminophors of p-terphenyl containing different amounts of perylene followed by spectral characterization techniques viz. XRD, SEM, TGA-DSC, UV-Visible spectroscopy, thermo-electrical conductivity, fluorescence spectroscopy, fluorescence life time spectroscopy and temperature dependent fluorescence. X-ray diffraction profiles of the doped p-terphenyl reveal well-defined and sharp peaks indicate homogeneity and crystallinity. The SEM micrograph of pure p-terphenyl exhibit flakes like grains and then compact and finally gets separately with perylene amounts. The observed results indicate that closed packed crystal structures of doped p-terphenyl during crystal formation. The band gaps estimated from UV-visible spectroscopy decreased from 5.20 to 4.10 eV, while thermo-electrical conductivity increases with perylene content. The fluorescence spectra showed partial quenching of p-terphenyl fluorescence and simultaneously sensitization of perylene fluorescence at the excitation wavelength of p-terphenyl (290 nm) due to excitation energy transfer from p-terphenyl to perylene. The observed sensitization results are in harmony with intense blue color seen in fluorescence microscopy images and has high demand in scintillation process.

High contrast two-photon imaging of fingermarks

NASA Astrophysics Data System (ADS)

Stoltzfus, Caleb R.; Rebane, Aleksander

2016-04-01

Optically-acquired fingermarks are widely used as evidence across law enforcement agencies as well as in the courts of law. A common technique for visualizing latent fingermarks on nonporous surfaces consists of cyanoacrylate fuming of the fingerprint material, followed by impregnation with a fluorescent dye, which under ultra violet (UV) illumination makes the fingermarks visible and thus accessible for digital recording. However, there exist critical circumstances, when the image quality is compromised due to high background scattering, high auto-fluorescence of the substrate material, or other detrimental photo-physical and photo-chemical effects such as light-induced damage to the sample. Here we present a novel near-infrared (NIR), two-photon induced fluorescence imaging modality, which significantly enhances the quality of the fingermark images, especially when obtained from highly reflective and/or scattering surfaces, while at the same time reducing photo-damage to sensitive forensic samples.

Integrated smartphone imaging of quantum dot photoluminescence and Förster resonance energy transfer

NASA Astrophysics Data System (ADS)

Petryayeva, Eleonora; Algar, W. Russ

2015-06-01

Smartphones and other mobile devices are emerging as promising analytical platforms for point-of-care diagnostics, particularly when combined with nanotechnology. For example, we have shown that the optical properties of semiconductor quantum dots (QDs) are well suited to photoluminescence (PL) detection with a smartphone camera. However, this previous work has utilized an external excitation source for interrogation of QD PL. In this proceeding, we demonstrate that the white-light LED photographic flashes built into smartphones can be optically filtered to yield blue light suitable for excitation of QD PL. Measurements were made by recording video with filtered flash illumination and averaging the frames of the video to obtain images with good signal-to-background ratios. These images permitted detection of green-emitting and red-emitting QDs at levels comparable to those possible with excitation using an external long-wave UV lamp. The optical properties of QDs proved to be uniquely suited to smartphone PL imaging, exhibiting emission that was 1-2 orders magnitude brighter than that of common fluorescent dyes under the same conditions. Excitation with the smartphone flash was also suitable for imaging of FRET between green-emitting QD donors and Alexa Fluor 555 (A555) fluorescent dye acceptors. No significant difference in FRET imaging capability was observed between excitation with the smartphone flash and a long-wave UV lamp. Although the smartphone flash did have some disadvantages compared to an external UV lamp, these disadvantages are potentially offset by the benefit of having excitation and detection integrated into the smartphone.

Caging and Photoactivation in Single-Molecule Förster Resonance Energy Transfer Experiments

PubMed Central

2017-01-01

Caged organic fluorophores are established tools for localization-based super-resolution imaging. Their use relies on reversible deactivation of standard organic fluorophores by chemical reduction or commercially available caged dyes with ON switching of the fluorescent signal by ultraviolet (UV) light. Here, we establish caging of cyanine fluorophores and caged rhodamine dyes, i.e., chemical deactivation of fluorescence, for single-molecule Förster resonance energy transfer (smFRET) experiments with freely diffusing molecules. They allow temporal separation and sorting of multiple intramolecular donor–acceptor pairs during solution-based smFRET. We use this “caged FRET” methodology for the study of complex biochemical species such as multisubunit proteins or nucleic acids containing more than two fluorescent labels. Proof-of-principle experiments and a characterization of the uncaging process in the confocal volume are presented. These reveal that chemical caging and UV reactivation allow temporal uncoupling of convoluted fluorescence signals from, e.g., multiple spectrally similar donor or acceptor molecules on nucleic acids. We also use caging without UV reactivation to remove unwanted overlabeled species in experiments with the homotrimeric membrane transporter BetP. We finally outline further possible applications of the caged FRET methodology, such as the study of weak biochemical interactions, which are otherwise impossible with diffusion-based smFRET techniques because of the required low concentrations of fluorescently labeled biomolecules. PMID:28362086

Fluorescence imaging to quantify crop residue cover

NASA Technical Reports Server (NTRS)

Daughtry, C. S. T.; Mcmurtrey, J. E., III; Chappelle, E. W.

1994-01-01

Crop residues, the portion of the crop left in the field after harvest, can be an important management factor in controlling soil erosion. Methods to quantify residue cover are needed that are rapid, accurate, and objective. Scenes with known amounts of crop residue were illuminated with long wave ultraviolet (UV) radiation and fluorescence images were recorded with an intensified video camera fitted with a 453 to 488 nm band pass filter. A light colored soil and a dark colored soil were used as background for the weathered soybean stems. Residue cover was determined by counting the proportion of the pixels in the image with fluorescence values greater than a threshold. Soil pixels had the lowest gray levels in the images. The values of the soybean residue pixels spanned nearly the full range of the 8-bit video data. Classification accuracies typically were within 3(absolute units) of measured cover values. Video imaging can provide an intuitive understanding of the fraction of the soil covered by residue.

Portable multispectral fluorescence imaging system for food safety applications

NASA Astrophysics Data System (ADS)

Lefcourt, Alan M.; Kim, Moon S.; Chen, Yud-Ren

2004-03-01

Fluorescence can be a sensitive method for detecting food contaminants. Of particular interest is detection of fecal contamination as feces is the source of many pathogenic organisms. Feces generally contain chlorophyll a and related compounds due to ingestion of plant materials, and these compounds can readily be detected using fluorescence techniques. Described is a fluorescence-imaging system consisting primarily of a UV light source, an intensified camera with a six-position filter wheel, and software for controlling the system and automatically analyzing the resulting images. To validate the system, orchard apples artificially contaminated with dairy feces were used in a "hands-on" public demonstration. The contamination sites were easily identified using automated edge detection and threshold detection algorithms. In addition, by applying feces to apples and then washing sets of apples at hourly intervals, it was determined that five h was the minimum contact time that allowed identification of the contamination site after the apples were washed. There are many potential uses for this system, including studying the efficacy of apple washing systems.

Deep UV laser-induced fluorescence detection of unlabeled drugs and proteins in microchip electrophoresis.

PubMed

Schulze, Philipp; Ludwig, Martin; Kohler, Frank; Belder, Detlev

2005-03-01

Deep UV fluorescence detection at 266-nm excitation wavelength has been realized for sensitive detection in microchip electrophoresis. For this purpose, an epifluorescence setup was developed enabling the coupling of a deep UV laser into a commercial fluorescence microscope. Deep UV laser excitation utilizing a frequency quadrupled pulsed laser operating at 266 nm shows an impressive performance for native fluorescence detection of various compounds in fused-silica microfluidic devices. Aromatic low molecular weight compounds such as serotonin, propranolol, a diol, and tryptophan could be detected at low-micromolar concentrations. Deep UV fluorescence detection was also successfully employed for the detection of unlabeled basic proteins. For this purpose, fused-silica chips dynamically coated with hydroxypropylmethyl cellulose were employed to suppress analyte adsorption. Utilizing fused-silica chips permanently coated with poly(vinyl alcohol), it was also possible to separate and detect egg white chicken proteins. These data show that deep UV fluorescence detection significantly widens the application range of fluorescence detection in chip-based analysis techniques.

Interaction of amino acid-functionalized silver nanoparticles and Candida albicans polymorphs: A deep-UV fluorescence imaging study.

PubMed

Dojčilović, Radovan; Pajović, Jelena D; Božanić, Dušan K; Bogdanović, Una; Vodnik, Vesna V; Dimitrijević-Branković, Suzana; Miljković, Miona G; Kaščaková, Slavka; Réfrégiers, Matthieu; Djoković, Vladimir

2017-07-01

The interaction of the tryptophan functionalized Ag nanoparticles and live Candida albicans cells was studied by synchrotron excitation deep-ultraviolet (DUV) fluorescence imaging at the DISCO beamline of Synchrotron SOLEIL. DUV imaging showed that incubation of the fungus with functionalized nanoparticles results in significant increase in the fluorescence signal. The analysis of the images revealed that the interaction of the nanoparticles with (pseudo)hyphae polymorphs of the diploid fungus was less pronounced than in the case of yeast cells or budding spores. The changes in the intensity of the fluorescence signals of the cells after incubation were followed in [327-353nm] and [370-410nm] spectral ranges that correspond to the fluorescence of tryptophan in non-polar and polar environment, respectively. As a consequence of the environmental sensitivity of the silver-tryptophan fluorescent nanoprobe, we were able to determine the possible accumulation sites of the nanoparticles. The analysis of the intensity decay kinetics showed that the photobleaching effects were more pronounced in the case of the functionalized nanoparticle treated cells. The results of time-integrated emission in the mentioned spectral ranges suggested that the nanoparticles penetrate the cells, but that the majority of the nanoparticles attach to the cells' surfaces. Copyright © 2017 Elsevier B.V. All rights reserved.

Ultraviolet /UV/ sensitive phosphors for silicon imaging detectors

NASA Technical Reports Server (NTRS)

Viehmann, W.; Cowens, M. W.; Butner, C. L.

1981-01-01

The fluorescence properties of UV sensitive organic phosphors and the radiometric properties of phosphor coated silicon detectors in the VUV, UV, and visible wavelengths are described. With evaporated films of coronene and liumogen, effective quantum efficiencies of up to 20% have been achieved on silicon photodiodes in the vacuum UV. With thin films of methylmethacrylate (acrylic), which are doped with organic laser dyes and deposited from solution, detector quantum efficiencies of the order of 15% for wavelengths of 120-165 nm and of 40% for wavelengths above 190 nm have been obtained. The phosphor coatings also act as antireflection coatings and thereby enhance the response of coated devices throughout the visible and near IR.

Developing LED UV fluorescence sensors for online monitoring DOM and predicting DBPs formation potential during water treatment.

PubMed

Li, Wen-Tao; Jin, Jing; Li, Qiang; Wu, Chen-Fei; Lu, Hai; Zhou, Qing; Li, Ai-Min

2016-04-15

Online monitoring dissolved organic matter (DOM) is urgent for water treatment management. In this study, high performance size exclusion chromatography with multi-UV absorbance and multi-emission fluorescence scans were applied to spectrally characterize samples from 16 drinking water sources across Yangzi River and Huai River Watersheds. The UV absorbance indices at 254 nm and 280 nm referred to the same DOM components and concentration, and the 280 nm UV light could excite both protein-like and humic-like fluorescence. Hence a novel UV fluorescence sensor was developed out using only one UV280 light-emitting diode (LED) as light source. For all samples, enhanced coagulation was mainly effective for large molecular weight biopolymers; while anion exchange further substantially removed humic substances. During chlorination tests, UVA280 and UVA254 showed similar correlations with yields of disinfection byproducts (DBPs); the humic-like fluorescence obtained from LED sensors correlated well with both trihalomethanes and haloacetic acids yields, while the correlation between protein-like fluorescence and trihalomethanes was relatively poor. Anion exchange exhibited more reduction of DBPs yields as well as UV absorbance and fluorescence signals than enhanced coagulation. The results suggest that the LED UV fluorescence sensors are very promising for online monitoring DOM and predicting DBPs formation potential during water treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Development of UV-curable liquid for in-liquid fluorescence alignment in ultraviolet nanoimprint lithography

NASA Astrophysics Data System (ADS)

Ochiai, Kento; Kikuchi, Eri; Ishito, Yota; Kumagai, Mari; Nakamura, Takahiro; Nakagawa, Masaru

2018-06-01

We studied a fluorescent UV-curable resin suitable for fluorescence alignment in UV nanoimprinting. The addition of a cationic fluorescent dye caused radical photopolymerization of a UV-curable resin by exposure to visible excitation light for fluorescence microscope observation. The microscope observation of a resin film prepared by pressing resin droplets on a silica substrate with a fluorinated silica superstrate revealed that the cationic dye molecules were preferably adsorbed onto the silica surface. It was indicated that the dye molecules concentrated on the silica surface may cause the photocuring. A nonionic fluorescent dye was selected owing to its low polar symmetrical structure and its solubility parameter close to monomers. The fluorescent UV-curable resin with the nonionic dye showed uncured stability to exposure to visible excitation light for 30 min with a light intensity of 8.5 mW cm‑2 detected at 530 nm.

Portable multispectral imaging system for oral cancer diagnosis

NASA Astrophysics Data System (ADS)

Hsieh, Yao-Fang; Ou-Yang, Mang; Lee, Cheng-Chung

2013-09-01

This study presents the portable multispectral imaging system that can acquire the image of specific spectrum in vivo for oral cancer diagnosis. According to the research literature, the autofluorescence of cells and tissue have been widely applied to diagnose oral cancer. The spectral distribution is difference for lesions of epithelial cells and normal cells after excited fluorescence. We have been developed the hyperspectral and multispectral techniques for oral cancer diagnosis in three generations. This research is the third generation. The excited and emission spectrum for the diagnosis are acquired from the research of first generation. The portable system for detection of oral cancer is modified for existing handheld microscope. The UV LED is used to illuminate the surface of oral cavity and excite the cells to produce fluorescent. The image passes through the central channel and filters out unwanted spectrum by the selection of filter, and focused by the focus lens on the image sensor. Therefore, we can achieve the specific wavelength image via fluorescence reaction. The specificity and sensitivity of the system are 85% and 90%, respectively.

Non-destructive evaluation of UV pulse laser-induced damage performance of fused silica optics.

PubMed

Huang, Jin; Wang, Fengrui; Liu, Hongjie; Geng, Feng; Jiang, Xiaodong; Sun, Laixi; Ye, Xin; Li, Qingzhi; Wu, Weidong; Zheng, Wanguo; Sun, Dunlu

2017-11-24

The surface laser damage performance of fused silica optics is related to the distribution of surface defects. In this study, we used chemical etching assisted by ultrasound and magnetorheological finishing to modify defect distribution in a fused silica surface, resulting in fused silica samples with different laser damage performance. Non-destructive test methods such as UV laser-induced fluorescence imaging and photo-thermal deflection were used to characterize the surface defects that contribute to the absorption of UV laser radiation. Our results indicate that the two methods can quantitatively distinguish differences in the distribution of absorptive defects in fused silica samples subjected to different post-processing steps. The percentage of fluorescence defects and the weak absorption coefficient were strongly related to the damage threshold and damage density of fused silica optics, as confirmed by the correlation curves built from statistical analysis of experimental data. The results show that non-destructive evaluation methods such as laser-induced fluorescence and photo-thermal absorption can be effectively applied to estimate the damage performance of fused silica optics at 351 nm pulse laser radiation. This indirect evaluation method is effective for laser damage performance assessment of fused silica optics prior to utilization.

Norcyanine dyes with benzo[c,d]indolium moiety: Spectral sensitivity with pH change for fluorescence pH imaging in living cells.

PubMed

Guan, Li; Liu, Qi; Zhang, Borui; Wang, Lanying

2017-01-01

Fluorescence pH imaging in living cells is a rapidly expanding research direction, however, it relies on the development of pH-sensitive fluorescent imaging agents. Here four norcyanine dyes with benzo[c,d]indolium moiety, exhibiting high spectral sensitivity with pH changes, were synthesized for fluorescence pH imaging in living cells, and characterized by 1 H NMR, 13 C NMR, IR, UV-Vis and HRMS. The investigation of their spectral properties in methanol and water showed that the absorption and emission maxima were in the region 488-618nm and 583-651nm, respectively, and four dyes exhibited high photostability. The pH spectral titrations showed that selective dye D1 had pH-dependent absorption spectral changes within the pH range of 2.4 to 9.4, and high fluorescent spectral sensitivity at pH5.0-8.0, with a pK a of 5.0. A cell association study indicated that dye D1 exhibited no or mild cytotoxicity at the application dose and duration, and could be accumulated in cells and mainly distributed in the cytoplasm, giving red fluorescence imaging. In particular, dye D1 could achieve pH-dependent fluorescence imaging in living cells with the increase of pH from 3.0 to 8.0, at excitation wavelength of 543nm and receiving wavelength of 655-755nm, which was valuable for studying the weak acidic, neutral and weak alkaline biological tissue compartments. Copyright © 2016 Elsevier B.V. All rights reserved.

Oxygen-doped carbon nanotubes for near-infrared fluorescent labels and imaging probes.

PubMed

Iizumi, Yoko; Yudasaka, Masako; Kim, Jaeho; Sakakita, Hajime; Takeuchi, Tsukasa; Okazaki, Toshiya

2018-04-19

Chemical modification of carbon nanotube surface can controllably modulate their optical properties. Here we report a simple and effective synthesis method of oxygen-doped single-walled carbon nanotubes (o-SWCNTs), in which a thin film of SWCNTs is just irradiated under the UV light for a few minutes in air. By using this method, the epoxide-type oxygen-adducts (ep-SWCNTs) were produced in addition to the ether-type oxygen-adducts (eth-SWCNTs). The Treated (6, 5) ep-SWCNTs show a red-shifted luminescence at ~1280 nm, which corresponds to the most transparent regions for bio-materials. Immunoassay, fluorescence vascular angiography and observation of the intestinal contractile activity of mice were demonstrated by using the produced o-SWCNTs as infrared fluorescent labels and imaging agents.

Hyperspectral imaging for detection of black tip damage in wheat kernels

NASA Astrophysics Data System (ADS)

Delwiche, Stephen R.; Yang, I.-Chang; Kim, Moon S.

2009-05-01

A feasibility study was conducted on the use of hyperspectral imaging to differentiate sound wheat kernels from those with the fungal condition called black point or black tip. Individual kernels of hard red spring wheat were loaded in indented slots on a blackened machined aluminum plate. Damage conditions, determined by official (USDA) inspection, were either sound (no damage) or damaged by the black tip condition alone. Hyperspectral imaging was separately performed under modes of reflectance from white light illumination and fluorescence from UV light (~380 nm) illumination. By cursory inspection of wavelength images, one fluorescence wavelength (531 nm) was selected for image processing and classification analysis. Results indicated that with this one wavelength alone, classification accuracy can be as high as 95% when kernels are oriented with their dorsal side toward the camera. It is suggested that improvement in classification can be made through the inclusion of multiple wavelength images.

Interrogating Surface Functional Group Heterogeneity of Activated Thermoplastics Using Super-Resolution Fluorescence Microscopy.

PubMed

ONeil, Colleen E; Jackson, Joshua M; Shim, Sang-Hee; Soper, Steven A

2016-04-05

We present a novel approach for characterizing surfaces utilizing super-resolution fluorescence microscopy with subdiffraction limit spatial resolution. Thermoplastic surfaces were activated by UV/O3 or O2 plasma treatment under various conditions to generate pendant surface-confined carboxylic acids (-COOH). These surface functional groups were then labeled with a photoswitchable dye and interrogated using single-molecule, localization-based, super-resolution fluorescence microscopy to elucidate the surface heterogeneity of these functional groups across the activated surface. Data indicated nonuniform distributions of these functional groups for both COC and PMMA thermoplastics with the degree of heterogeneity being dose dependent. In addition, COC demonstrated relative higher surface density of functional groups compared to PMMA for both UV/O3 and O2 plasma treatment. The spatial distribution of -COOH groups secured from super-resolution imaging were used to simulate nonuniform patterns of electroosmotic flow in thermoplastic nanochannels. Simulations were compared to single-particle tracking of fluorescent nanoparticles within thermoplastic nanoslits to demonstrate the effects of surface functional group heterogeneity on the electrokinetic transport process.

Detecting Kerogen as a Biosignature Using Colocated UV Time-Gated Raman and Fluorescence Spectroscopy.

PubMed

Shkolyar, Svetlana; Eshelman, Evan J; Farmer, Jack D; Hamilton, David; Daly, Michael G; Youngbull, Cody

2018-04-01

The Mars 2020 mission will analyze samples in situ and identify any that could have preserved biosignatures in ancient habitable environments for later return to Earth. Highest priority targeted samples include aqueously formed sedimentary lithologies. On Earth, such lithologies can contain fossil biosignatures as aromatic carbon (kerogen). In this study, we analyzed nonextracted kerogen in a diverse suite of natural, complex samples using colocated UV excitation (266 nm) time-gated (UV-TG) Raman and laser-induced fluorescence spectroscopies. We interrogated kerogen and its host matrix in samples to (1) explore the capabilities of UV-TG Raman and fluorescence spectroscopies for detecting kerogen in high-priority targets in the search for possible biosignatures on Mars; (2) assess the effectiveness of time gating and UV laser wavelength in reducing fluorescence in Raman spectra; and (3) identify sample-specific issues that could challenge rover-based identifications of kerogen using UV-TG Raman spectroscopy. We found that ungated UV Raman spectroscopy is suited to identify diagnostic kerogen Raman bands without interfering fluorescence and that UV fluorescence spectroscopy is suited to identify kerogen. These results highlight the value of combining colocated Raman and fluorescence spectroscopies, similar to those obtainable by SHERLOC on Mars 2020, to strengthen the confidence of kerogen detection as a potential biosignature in complex natural samples. Key Words: Raman spectroscopy-Laser-induced fluorescence spectroscopy-Mars Sample Return-Mars 2020 mission-Kerogen-Biosignatures. Astrobiology 18, 431-453.

Synthesis and Live-Cell Imaging of Fluorescent Sterols for Analysis of Intracellular Cholesterol Transport.

PubMed

Modzel, Maciej; Lund, Frederik W; Wüstner, Daniel

2017-01-01

Cellular cholesterol homeostasis relies on precise control of the sterol content of organelle membranes. Obtaining insight into cholesterol trafficking pathways and kinetics by live-cell imaging relies on two conditions. First, one needs to develop suitable analogs that resemble cholesterol as closely as possible with respect to their biophysical and biochemical properties. Second, the cholesterol analogs should have good fluorescence properties. This interferes, however, often with the first requirement, such that the imaging instrumentation must be optimized to collect photons from suboptimal fluorophores, but good cholesterol mimics, such as the intrinsically fluorescent sterols, cholestatrienol (CTL) or dehydroergosterol (DHE). CTL differs from cholesterol only in having two additional double bonds in the ring system, which is why it is slightly fluorescent in the ultraviolet (UV). In the first part of this protocol, we describe how to synthesize and image CTL in living cells relative to caveolin, a structural component of caveolae. In the second part, we explain in detail how to perform time-lapse experiments of commercially available BODIPY-tagged cholesterol (TopFluor-cholesterol ® ; TF-Chol) in comparison to DHE. Finally, using two-photon time-lapse imaging data of TF-Chol, we demonstrate how to use our imaging toolbox SpatTrack for tracking sterol rich vesicles in living cells over time.

System and method for controlling depth of imaging in tissues using fluorescence microscopy under ultraviolet excitation following staining with fluorescing agents

DOE Office of Scientific and Technical Information (OSTI.GOV)

Demos, Stavros; Levenson, Richard

The present disclosure relates to a method for analyzing tissue specimens. In one implementation the method involves obtaining a tissue sample and exposing the sample to one or more fluorophores as contrast agents to enhance contrast of subcellular compartments of the tissue sample. The tissue sample is illuminated by an ultraviolet (UV) light having a wavelength between about 200 nm to about 400 nm, with the wavelength being selected to result in penetration to only a specified depth below a surface of the tissue sample. Inter-image operations between images acquired under different imaging parameters allow for improvement of the imagemore » quality via removal of unwanted image components. A microscope may be used to image the tissue sample and provide the image to an image acquisition system that makes use of a camera. The image acquisition system may create a corresponding image that is transmitted to a display system for processing and display.« less

In vitro evidence for UV-protection of the eye by the corneal epithelium mediated by the cytoplasmic protein, RNA, and ascorbate.

PubMed

Ringvold, A

1997-10-01

(1) To evaluate the effect of ribonucleic acid (RNA) and ascorbate on UV-absorption and their impact on ultraviolet-induced (UV) fluorescence from various proteins, and (2) to compare RNA and DNA reduction of protein fluorescence. These informations will be useful for later work on the UV-filtering effect of the corneal epithelium. Spectrophotometry and spectrofluorimetry. (1) RNA and ascorbate caused a significant UV-absorption and reduced the fluorescence from various water-soluble proteins, the degree of reduction varying independently from one protein to the other. (2) RNA and DNA showed protein fluorescence reduction of roughly the same order. The results are discussed both in the context of UV-protection of the cell nucleus in general, and the possible UV-filtering effect for the eye of bovine corneal epithelium.

Polarized fluorescence for skin cancer diagnostic with a multi-aperture camera

NASA Astrophysics Data System (ADS)

Kandimalla, Haripriya; Ramella-Roman, Jessica C.

2008-02-01

Polarized fluorescence has shown some promising results in assessment of skin cancer margins. Researchers have used tetracycline and cross polarization imaging for nonmelanoma skin cancer demarcation as well as investigating endogenous skin polarized fluorescence. In this paper we present a new instrument for polarized fluorescence imaging, able to calculate the full fluorescence Stokes vector in one snapshot. The core of our system is a multi-aperture camera constructed with a two by two lenslet array. Three of the lenses have polarizing elements in front of them, oriented at 0°, + 45°and 90° with respect to light source polarization. A flash lamp combined with a polarizer parallel to the source-camera-sample plane and a UV filter is used as an excitation source. A blue filter in front of the camera system is used to collect only the fluorescent emission of interest and filter out the incident light. In-vitro tests of endogenous and exogenous polarized fluorescence on collagen rich material like bovine tendon were performed and Stokes vector of polarized fluorescence calculated. The system has the advantage of eliminating moving artifacts with the collection of different polarization states and stoke vector in a single snap shot.

As Light Meets Matter: Art under Scrutiny.

ERIC Educational Resources Information Center

Del Federico, Eleonora; Diver, Steven; Konaklieva, Monika; Ludescher, Richard

2002-01-01

Presents a story on an investigation of the painter of an artwork (who is suspected to be Cezanne) that uses UV spectroscopy, IR spectrum, X-ray fluorescence, and luminescence images. Leaves the story open-ended as to whether the painting is authentic. Includes teaching notes and suggestions for classroom management. (YDS)

Combined Contrast-Enhanced MRI and Fluorescence Molecular Tomography for Breast Tumor Imaging

DTIC Science & Technology

2009-03-01

analysis. Reportable outcomes This fellowship led to three first peer-reviewed publications, three oral presentations and two poster presentations...imaging applications. A design goal of the system pre- sented here was the development of a multiwavelength ca- pable detection system with a large...smaller given the availability of information at additional wavelengths. The extinction spectrum of LuTex was measured directly using a Cary 50 UV -Vis

High-speed fluorescent thermal imaging of quench propagation in high temperature superconductor tapes

NASA Astrophysics Data System (ADS)

Gyuráki, Roland; Sirois, Frédéric; Grilli, Francesco

2018-07-01

Fluorescent microthermographic imaging, a method using rare-earth fluorescent coatings with temperature dependent light emission, was used for quench investigation in high temperature superconductors (HTS). A fluorophore was embedded in a polymer matrix and used as a coating on top of an HTS tape, while being excited with UV light and recorded with a high-speed camera. Simultaneously, the tape was pulsed with high amplitude, short duration DC current, and brought to quench with the help of a localised defect. The Joule heating during a quench influences the fluorescent light intensity emitted from the coating, and by recording the local variations in this intensity over time, the heating of the tape can be visualised and the developed temperatures can be calculated. In this paper, the fluorophore europium tris[3-(trifluoromethylhydroxymethylene)-(+)-camphorate] (EuTFC) provided sufficient temperature sensitivity and a usable temperature range from 77-260 K. With the help of 2500 image captures per second, the normal zone development was imaged in a 20 μm copper stabilised HTS tape held in a liquid nitrogen bath, and using a calibration curve, the temperatures reached during the quench have been calculated.

X ray sensitive area detection device

NASA Technical Reports Server (NTRS)

Carter, Daniel C. (Inventor); Witherow, William K. (Inventor); Pusey, Marc L. (Inventor); Yost, Vaughn H. (Inventor)

1990-01-01

A radiation sensitive area detection device is disclosed which comprises a phosphor-containing film capable of receiving and storing an image formed by a pattern of incoming x rays, UV, or other radiation falling on the film. The device is capable of fluorescing in response to stimulation by a light source in a manner directly proportional to the stored radiation pattern. The device includes: (1) a light source capable of projecting light or other appropriate electromagnetic wave on the film so as to cause it to fluoresce; (2) a means to focus the fluoresced light coming from the phosphor-containing film after light stimulation; and (3) at least one charged coupled detector or other detecting element capable of receiving and digitizing the pattern of fluoresced light coming from the phosphor-containing film. The device will be able to generate superior x ray images of high resolution from a crystal or other sample and will be particularly advantageous in that instantaneous near-real-time images of rapidly deteriorating samples can be obtained. Furthermore, the device can be made compact and sturdy, thus capable of carrying out x ray or other radiation imaging under a variety of conditions, including those experienced in space.

Two-dimensional imaging of molecular hydrogen in H2-air diffusion flames using two-photon laser-induced fluorescence

NASA Technical Reports Server (NTRS)

Lempert, W.; Kumar, V.; Glesk, I.; Miles, R.; Diskin, G.

1991-01-01

The use of a tunable ArF laser at 193.26 nm to record simultaneous single-laser-shot, planar images of molecular hydrogen and hot oxygen in a turbulent H2-air diffusion flame. Excitation spectra of fuel and oxidant-rich flame zones confirm a partial overlap of the two-photon H2 and single-photon O2 Schumann-Runge absorption bands. UV Rayleigh scattering images of flame structure and estimated detection limits for the H2 two-photon imaging are also presented.

A coumarin based Schiff base probe for selective fluorescence detection of Al3 + and its application in live cell imaging

NASA Astrophysics Data System (ADS)

Sen, Bhaskar; Sheet, Sanjoy Kumar; Thounaojam, Romita; Jamatia, Ramen; Pal, Amarta Kumar; Aguan, Kripamoy; Khatua, Snehadrinarayan

2017-02-01

A new coumarin based Schiff base compound, CSB-1 has been synthesized to detect metal ion based on the chelation enhanced fluorescence (CHEF). The cation binding properties of CSB-1 was thoroughly examined in UV-vis and fluorescence spectroscopy. In fluorescence spectroscopy the compound showed high selectivity toward Al3 + ion and the Al3 + can be quantified in mixed aqueous buffer solution (MeOH: 0.01 M HEPES Buffer; 9:1; v/v) at pH 7.4 as well as in BSA media. The fluorescence intensity of CSB-1 was enhanced by 24 fold after addition of only five equivalents of Al3 +. The fluorescence titration of CSB-1 with Al3 + in mixed aqueous buffer afforded a binding constant, Ka = (1.06 ± 0.2) × 104 M- 1. The colour change from light yellow to colourless and the appearance of blue fluorescence, which can be observed by the naked eye, provides a real-time method for Al3 + sensing. Further the live cell imaging study indicated that the detection of intracellular Al3 + ions are also readily possible in living cell.

Synthesis and characterization of photoswitchable fluorescent silica nanoparticles.

PubMed

Fölling, Jonas; Polyakova, Svetlana; Belov, Vladimir; van Blaaderen, Alfons; Bossi, Mariano L; Hell, Stefan W

2008-01-01

We have designed and synthesized a new functional (amino reactive) highly efficient fluorescent molecular switch (FMS) with a photochromic diarylethene and a rhodamine fluorescent dye. The reactive group in this FMS -N-hydroxysuccinimide ester- allows selective labeling of amino containing molecules or other materials. In ethanolic solutions, the compound displays a large fluorescent quantum yield of 52 % and a large fluorescence modulation ratio (94 %) between two states that may be interconverted with red and near-UV light. Silica nanoparticles incorporating the new FMS were prepared and characterized, and their spectroscopic and switching properties were also studied. The dye retained its properties after the incorporation into the silica, thereby allowing light-induced reversible high modulation of the fluorescence signal of a single particle for up to 60 cycles, before undergoing irreversible photobleaching. Some applications of these particles in fluorescence microscopy are also demonstrated. In particular, subdiffraction images of nanoparticles were obtained, in the focal plane of a confocal microscope.

Detecting Kerogen as a Biosignature Using Co-located UV Time-Gated Raman and Fuorescence Spectroscopy

NASA Astrophysics Data System (ADS)

Shkolyar, S.; Eshelman, E.; Farmer, J. D.; Hamilton, D.; Daly, M. G.; Youngbull, C.

2017-12-01

The Mars 2020 mission will analyze samples in situ and identify any that could have preserved biosignatures in ancient habitable environments for later return to Earth. Highest-priority targeted samples include aqueously formed sedimentary lithologies containing fossil biosignatures as aromatic carbon (kerogen). In this study, we analyze non-extracted, naturally preserved kerogen in a diverse suite of realistic Mars analogs using combined UV excitation time-gated (UV-TG) Raman and laser-induced fluorescence spectroscopy. We interrogated kerogen and its host matrix in samples to: (1) explore the capabilities of UV-TG Raman and fluorescence spectroscopy for detecting kerogen in high-priority targets in the search for a Martian fossil record; (2) assess the effectiveness of time-gating and UV laser wavelength in reducing fluorescence; and (3) identify sample-specific issues which could challenge rover-based identifications of kerogen using UV-TG Raman spectroscopy. We found that ungated UV Raman is suited to identify diagnostic kerogen Raman bands without interfering fluorescence and that fluorescence features indicating kerogen are detectable. These data highlight the value of using both co-located Raman and fluorescence data sets together to strengthen the confidence of kerogen detection as a potential biosignature and are obtainable by SHERLOC onboard Mars 2020.

Ultraviolet signals in birds are special.

PubMed

Hausmann, Franziska; Arnold, Kathryn E; Marshall, N Justin; Owens, Ian P F

2003-01-07

Recent behavioural experiments have shown that birds use ultraviolet (UV)-reflective and fluorescent plumage as cues in mate choice. It remains controversial, however, whether such UV signals play a special role in sexual communication, or whether they are part of general plumage coloration. We use a comparative approach to test for a general association between sexual signalling and either UV-reflective or fluorescent plumage. Among the species surveyed, 72% have UV colours and there is a significant positive association between UV reflectance and courtship displays. Among parrots (Psittaciformes), 68% of surveyed species have fluorescent plumage, and again there is a strong positive association between courtship displays and fluorescence. These associations are not artefacts of the plumage used in courtship displays, being generally more 'colourful' because there is no association between display and colours lacking UV reflectance or fluorescence. Equally, these associations are not phylogenetic artefacts because all results remain unchanged when families or genera, rather than species, are used as independent data points. We also find that, in parrots, fluorescent plumage is usually found adjacent to UV-reflective plumage. Using a simple visual model to examine one parrot, the budgerigar Melopsittacus undulatus, we show that the juxtaposition of UV-reflective and fluorescent plumage leads to a 25-fold increase in chromatic contrast to the budgerigar's visual system. Taken together, these results suggest that signals based on UV contrast are of special importance in the context of active sexual displays. We review briefly six hypotheses on why this may be the case: suitability for short-range signalling; high contrast with backgrounds; invisibility to predators; exploitation of pre-existing sensory biases; advertisement of feather structure; and amplification of behavioural signals.

Ultraviolet signals in birds are special.

PubMed Central

Hausmann, Franziska; Arnold, Kathryn E; Marshall, N Justin; Owens, Ian P F

2003-01-01

Recent behavioural experiments have shown that birds use ultraviolet (UV)-reflective and fluorescent plumage as cues in mate choice. It remains controversial, however, whether such UV signals play a special role in sexual communication, or whether they are part of general plumage coloration. We use a comparative approach to test for a general association between sexual signalling and either UV-reflective or fluorescent plumage. Among the species surveyed, 72% have UV colours and there is a significant positive association between UV reflectance and courtship displays. Among parrots (Psittaciformes), 68% of surveyed species have fluorescent plumage, and again there is a strong positive association between courtship displays and fluorescence. These associations are not artefacts of the plumage used in courtship displays, being generally more 'colourful' because there is no association between display and colours lacking UV reflectance or fluorescence. Equally, these associations are not phylogenetic artefacts because all results remain unchanged when families or genera, rather than species, are used as independent data points. We also find that, in parrots, fluorescent plumage is usually found adjacent to UV-reflective plumage. Using a simple visual model to examine one parrot, the budgerigar Melopsittacus undulatus, we show that the juxtaposition of UV-reflective and fluorescent plumage leads to a 25-fold increase in chromatic contrast to the budgerigar's visual system. Taken together, these results suggest that signals based on UV contrast are of special importance in the context of active sexual displays. We review briefly six hypotheses on why this may be the case: suitability for short-range signalling; high contrast with backgrounds; invisibility to predators; exploitation of pre-existing sensory biases; advertisement of feather structure; and amplification of behavioural signals. PMID:12590772

Development of multifunctional nanoparticles towards applications in non-invasive magnetic resonance imaging and axonal tracing.

PubMed

Du, Yan; Qin, Yubo; Li, Zizhen; Yang, Xiuying; Zhang, Jingchang; Westwick, Harrison; Tsai, Eve; Cao, Xudong

2017-12-01

A multifunctional nanobiomaterial has been developed by deliberately combining functions of superparamagnetism, fluorescence, and axonal tracing into one material. Superparamagnetic iron oxide nanoparticles were first synthesized and coated with a silica layer to prevent emission quenching through core-dye interactions; a fluorescent molecule, fluorescein isothiocyanate, was doped inside second layer of silica shell to improve photo-stability and to enable further thiol functionalization. Subsequently, biotinylated dextran amine, a sensitive axonal tracing reagent, was immobilized on the thiol-functionalized nanoparticle surfaces. The resulting nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, X-ray diffraction, X-ray photoelectron spectroscopy, UV-Vis spectroscopy, magnetic resonance imaging and fluorescence confocal microscopy. In vitro cell experiments using both undifferentiated and differentiated Neuro-2a cells showed that the cells were able to take up the nanoparticles intracellularly and that the nanoparticles showed good biocompatibilities. In summary, this new material demonstrated promising performances for both optical and magnetic resonance imaging modalities, suggesting its promising potentials in applications such as in non-invasive imaging, particularly in neuronal tracing.

Molecular Hydrogen Fluorescence in IC 63

NASA Technical Reports Server (NTRS)

Andersson, B-G

2005-01-01

This grant has supported the acquisition, reduction and analysis of data targeting the structure and excitation of molecular hydrogen in the reflection nebula IC 63 and in particular the fluorescent emission seen in the UV. In addition to manpower for analyzing the FUSE data, the grant supported the (attempted) acquisition of supporting ground-based data. We proposed for and received observing time for two sets of ground based, data; narrow band imaging ([S II], [O III) at KPNO (July 2002; Observer: Burgh) and imaging spectro-photometry of several of the near-infrared rotation-vibration lines of H2 at the IRTF (October 2003; Observer: Andersson). Unfortunately, both of these runs were failures, primarily because of bad weather, and did not result in any useful data. We combined the FUSE observations with rocket borne observations of the star responsible for exciting the H2 fluorescence in IC 63: gamma Cas, and with archival HUT observations of IC 63, covering the long-wavelength part of the molecular hydrogen fluorescence.

In vivo assessment of wound re-epithelialization by UV fluorescence excitation imaging

NASA Astrophysics Data System (ADS)

Wang, Ying; Ortega-Martinez, Antonio; Padilla-Martinez, Juan Pablo; Williams, Maura; Farinelli, William; Anderson, R. R.; Franco, Walfre

2017-02-01

Background and Objectives: We have previously demonstrated the efficacy of a non-invasive, non-contact, fast and simple but robust fluorescence imaging (u-FEI) method to monitor the healing of skin wounds in vitro. This system can image highly-proliferating cellular processes (295/340 nm excitation/emission wavelengths) to study epithelialization in a cultured wound model. The objective of the current work is to evaluate the suitability of u-FEI for monitoring wound re-epithelialization in vivo. Study Design: Full-thickness wounds were created in the tail of rats and imaged weekly using u-FEI at 295/340nm excitation/emission wavelengths. Histology was used to investigate the correlation between the spatial distribution and intensity of fluorescence and the extent of wound epithelialization. In addition, the expression of the nuclear protein Ki67 was used to confirm the association between the proliferation of keratinocyte cells and the intensity of fluorescence. Results: Keratinocytes forming neo-epidermis exhibited higher fluorescence intensity than the keratinocytes not involved in re-epithelialization. In full-thickness wounds the fluorescence first appeared at the wound edge where keratinocytes initiated the epithelialization process. Fluorescence intensity increased towards the center as the keratinocytes partially covered the wound. As the wound healed, fluorescence decreased at the edges and was present only at the center as the keratinocytes completely covered the wound at day 21. Histology demonstrated that changes in fluorescence intensity from the 295/340nm band corresponded to newly formed epidermis. Conclusions: u-FEI at 295/340nm allows visualization of proliferating keratinocyte cells during re-epithelialization of wounds in vivo, potentially providing a quantitative, objective and simple method for evaluating wound closure in the clinic.

Development of a safe ultraviolet camera system to enhance awareness by showing effects of UV radiation and UV protection of the skin (Conference Presentation)

NASA Astrophysics Data System (ADS)

Verdaasdonk, Rudolf M.; Wedzinga, Rosaline; van Montfrans, Bibi; Stok, Mirte; Klaessens, John; van der Veen, Albert

2016-03-01

The significant increase of skin cancer occurring in the western world is attributed to longer sun expose during leisure time. For prevention, people should become aware of the risks of UV light exposure by showing skin damage and the protective effect of sunscreen with an UV camera. An UV awareness imaging system optimized for 365 nm (UV-A) was develop using consumer components being interactive, safe and mobile. A Sony NEX5t camera was adapted to full spectral range. In addition, UV transparent lenses and filters were selected based on spectral characteristics measured (Schott S8612 and Hoya U-340 filters) to obtain the highest contrast for e.g. melanin spots and wrinkles on the skin. For uniform UV illumination, 2 facial tanner units were adapted with UV 365 nm black light fluorescent tubes. Safety of the UV illumination was determined relative to the sun and with absolute irradiance measurements at the working distance. A maximum exposure time over 15 minutes was calculate according the international safety standards. The UV camera was successfully demonstrated during the Dutch National Skin Cancer day and was well received by dermatologists and participating public. Especially, the 'black paint' effect putting sun screen on the face was dramatic and contributed to the awareness of regions on the face what are likely to be missed applying sunscreen. The UV imaging system shows to be promising for diagnostics and clinical studies in dermatology and potentially in other areas (dentistry and ophthalmology)

Detection of microbial biofilms on food processing surfaces: hyperspectral fluorescence imaging study

NASA Astrophysics Data System (ADS)

Jun, Won; Kim, Moon S.; Chao, Kaunglin; Lefcourt, Alan M.; Roberts, Michael S.; McNaughton, James L.

2009-05-01

We used a portable hyperspectral fluorescence imaging system to evaluate biofilm formations on four types of food processing surface materials including stainless steel, polypropylene used for cutting boards, and household counter top materials such as formica and granite. The objective of this investigation was to determine a minimal number of spectral bands suitable to differentiate microbial biofilm formation from the four background materials typically used during food processing. Ultimately, the resultant spectral information will be used in development of handheld portable imaging devices that can be used as visual aid tools for sanitation and safety inspection (microbial contamination) of the food processing surfaces. Pathogenic E. coli O157:H7 and Salmonella cells were grown in low strength M9 minimal medium on various surfaces at 22 +/- 2 °C for 2 days for biofilm formation. Biofilm autofluorescence under UV excitation (320 to 400 nm) obtained by hyperspectral fluorescence imaging system showed broad emissions in the blue-green regions of the spectrum with emission maxima at approximately 480 nm for both E. coli O157:H7 and Salmonella biofilms. Fluorescence images at 480 nm revealed that for background materials with near-uniform fluorescence responses such as stainless steel and formica cutting board, regardless of the background intensity, biofilm formation can be distinguished. This suggested that a broad spectral band in the blue-green regions can be used for handheld imaging devices for sanitation inspection of stainless, cutting board, and formica surfaces. The non-uniform fluorescence responses of granite make distinctions between biofilm and background difficult. To further investigate potential detection of the biofilm formations on granite surfaces with multispectral approaches, principal component analysis (PCA) was performed using the hyperspectral fluorescence image data. The resultant PCA score images revealed distinct contrast between biofilms and granite surfaces. This investigation demonstrated that biofilm formations on food processing surfaces, even for background materials with heterogeneous fluorescence responses, can be detected. Furthermore, a multispectral approach in developing handheld inspection devices may be needed to inspect surface materials that exhibit non-uniform fluorescence.

Hyperspectral Fluorescence and Reflectance Imaging Instrument

NASA Technical Reports Server (NTRS)

Ryan, Robert E.; O'Neal, S. Duane; Lanoue, Mark; Russell, Jeffrey

2008-01-01

The system is a single hyperspectral imaging instrument that has the unique capability to acquire both fluorescence and reflectance high-spatial-resolution data that is inherently spatially and spectrally registered. Potential uses of this instrument include plant stress monitoring, counterfeit document detection, biomedical imaging, forensic imaging, and general materials identification. Until now, reflectance and fluorescence spectral imaging have been performed by separate instruments. Neither a reflectance spectral image nor a fluorescence spectral image alone yields as much information about a target surface as does a combination of the two modalities. Before this system was developed, to benefit from this combination, analysts needed to perform time-consuming post-processing efforts to co-register the reflective and fluorescence information. With this instrument, the inherent spatial and spectral registration of the reflectance and fluorescence images minimizes the need for this post-processing step. The main challenge for this technology is to detect the fluorescence signal in the presence of a much stronger reflectance signal. To meet this challenge, the instrument modulates artificial light sources from ultraviolet through the visible to the near-infrared part of the spectrum; in this way, both the reflective and fluorescence signals can be measured through differencing processes to optimize fluorescence and reflectance spectra as needed. The main functional components of the instrument are a hyperspectral imager, an illumination system, and an image-plane scanner. The hyperspectral imager is a one-dimensional (line) imaging spectrometer that includes a spectrally dispersive element and a two-dimensional focal plane detector array. The spectral range of the current imaging spectrometer is between 400 to 1,000 nm, and the wavelength resolution is approximately 3 nm. The illumination system consists of narrowband blue, ultraviolet, and other discrete wavelength light-emitting-diode (LED) sources and white-light LED sources designed to produce consistently spatially stable light. White LEDs provide illumination for the measurement of reflectance spectra, while narrowband blue and UV LEDs are used to excite fluorescence. Each spectral type of LED can be turned on or off depending on the specific remote-sensing process being performed. Uniformity of illumination is achieved by using an array of LEDs and/or an integrating sphere or other diffusing surface. The image plane scanner uses a fore optic with a field of view large enough to provide an entire scan line on the image plane. It builds up a two-dimensional image in pushbroom fashion as the target is scanned across the image plane either by moving the object or moving the fore optic. For fluorescence detection, spectral filtering of a narrowband light illumination source is sometimes necessary to minimize the interference of the source spectrum wings with the fluorescence signal. Spectral filtering is achieved with optical interference filters and absorption glasses. This dual spectral imaging capability will enable the optimization of reflective, fluorescence, and fused datasets as well as a cost-effective design for multispectral imaging solutions. This system has been used in plant stress detection studies and in currency analysis.

Novel flashlamp-based time-resolved fluorescence microscope reduces autofluorescence for 30-fold contrast enhancement in environmental samples

NASA Astrophysics Data System (ADS)

Connally, Russell; Veal, Duncan; Piper, James A.

2003-07-01

The abundance of naturally fluorescing components (autofluorophors) encountered in environmentally sourced samples can greatly hinder the detection and identification of fluorescently labeled target using fluorescence microscopy. Time-resolved fluorescence microscopy (TRFM) is a technique that reduces the effects of autofluorescence through precisely controlled time delays. Lanthanide chelates have fluorescence lifetimes many orders of magnitude greater than typical autofluorophors, and persist in their luminescence long after autofluorescence has ceased. An intense short pulse of (UV) light is used to excite fluorescence in the sample and after a short delay period the longer persisting fluorescence from the chelate is captured with an image-intensified CCD camera. The choice of pulsed excitation source for TRFM has a large impact on the price and performance of the instrument. A flashlamp with a short pulse duration was selected for our instrument because of the high spectral energy in the UV region and short pulse length. However, flash output decays with an approximate lifetime of 18μs and the TRFM requires a long-lived chelate to ensure probe fluorescence is still visible after decay of the flash plasma. We synthesized a recently reported fluorescent chelate (BHHCT) and conjugated it to a monoclonal antibody directed against the water-borne parasite Giardia lamblia. Fluorescence lifetime of the construct was determined to be 339μs +/- 14μs and provided a 45-fold enhancement of labeled Giardia over background using a gate delay of 100μs. Despite the sub-optimal decay characteristics of the light pulse, flashlamps have many advantages compared to optical chopper wheels and modulated lasers. Their low cost, lack of vibration, ease of interface and small footprint are important factors to consider in TRFM design.

Design and demonstration of multimodal optical scanning microscopy for confocal and two-photon imaging

NASA Astrophysics Data System (ADS)

Chun, Wanhee; Do, Dukho; Gweon, Dae-Gab

2013-01-01

We developed a multimodal microscopy based on an optical scanning system in order to obtain diverse optical information of the same area of a sample. Multimodal imaging researches have mostly depended on a commercial microscope platform, easy to use but restrictive to extend imaging modalities. In this work, the beam scanning optics, especially including a relay lens, was customized to transfer broadband (400-1000 nm) lights to a sample without any optical error or loss. The customized scanning optics guarantees the best performances of imaging techniques utilizing the lights within the design wavelength. Confocal reflection, confocal fluorescence, and two-photon excitation fluorescence images were obtained, through respective implemented imaging channels, to demonstrate imaging feasibility for near-UV, visible, near-IR continuous light, and pulsed light in the scanning optics. The imaging performances for spatial resolution and image contrast were verified experimentally; the results were satisfactory in comparison with theoretical results. The advantages of customization, containing low cost, outstanding combining ability and diverse applications, will contribute to vitalize multimodal imaging researches.

Combining aminocyanine dyes with polyamide dendrons: a promising strategy for imaging in the near-infrared region.

PubMed

Ornelas, Cátia; Lodescar, Rachelle; Durandin, Alexander; Canary, James W; Pennell, Ryan; Liebes, Leonard F; Weck, Marcus

2011-03-21

Cyanine dyes are known for their fluorescence in the near-IR (NIR) region, which is desirable for biological applications. We report the synthesis of a series of aminocyanine dyes containing terminal functional groups such as acid, azide, and cyclooctyne groups for further functionalization through, for example, click chemistry. These aminocyanine dyes can be attached to polyfunctional dendrons by copper-catalyzed azide alkyne cycloaddition (CuAAC), strain-promoted azide alkyne cycloaddition (SPAAC), peptide coupling, or direct S(NR)1 reactions. The resulting dendron-dye conjugates were obtained in high yields and displayed high chemical stability and photostability. The optical properties of the new compounds were studied by UV/Vis and fluorescence spectroscopy. All compounds show large Stokes shifts and strong fluorescence in the NIR region with high quantum yields, which are optimal properties for in vivo optical imaging. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Photostability effect of silica nanoparticles encapsulated fluorescence dye

NASA Astrophysics Data System (ADS)

Ahmad, Atiqah; Zakaria, Nor Dyana; Razak, Khairunisak Abdul

2017-12-01

Fluorescence dyes are based on small organic molecules have become of interest in chemical biology and widely used for cell and intracellular imaging. However, fluorescence dyes have limitations such as photo bleaching, poor photochemical stability and has a short Stokes shift. It is less valuable for long-term cell tracking strategies and has very short lifetime. In order to overcome the problems, dye-incorporated nanomaterials become of interest. Nanomaterials encapsulation provides a protection layer around the fluorescence dye which improves the stability of fluorescence dye. In this study, silica nanoparticles encapsulated with 1,1%-dioctadecyl-3,3,3%,3%-tetramethylindocarbocyanine perchlorate (Dil) was successfully synthesised by using micelle entrapment method to investigate the effect of encapsulation of nanoparticles towards the properties of fluorescent dye. The synthesised nanoparticles (SiDil) was characterised by particle size analyser, Transmission Electron Microscopy (TEM), UV-Vis spectrometer and Fluorescent spectrometer. Observation using TEM showed spherical shape of nanoparticles with 53 nm diameter. Monodispersed and well nanoparticles distribution was confirmed by low polydispersity index of 0.063 obtained by particle size analyser. Furthermore, the photoluminescence properties of the SiDil were evaluated and compared with bare Dil dye. Both SiDil and bare Dil was radiated under 200 W of Halogen lamp for 60 minutes and the absorbance intensity was measured using UV-Vis spectrometer. The result showed more stable absorbance intensity for SiDil compared to bare Dil dye, which indicated that Si nanoparticles encapsulation improved the photostability property.

In-Field Diffuse Ultraviolet Spectroscopy and Imaging of the Stardust Sample Return Capsule

NASA Technical Reports Server (NTRS)

Pugel, D. Elizabeth; Stackpoole, Mairead; McNamara, Karen; Schwartz, C.; Warren, J.; Kontinos, Dean

2008-01-01

In-field diffuse Ultraviolet (UV) spectroscopy and imaging systems were developed for the purposes of evaluating the surface chemical composition of spacecraft thermal control coatings and materials. The investigation of these systems and the compilation of an associated UV reflectance and luminescence database were conducted using the Stardust Sample Return Capsule (SRC), located at the Johnson Space Center. Spectral responses of the surfaces of the Stardust forebody and aftbody in both reflectance and fluorescence modes were examined post-flight. In this paper, we report on two primary findings of in-field diffuse UV spectroscopy and imaging: (1) deduction of the thermal history of thermal control coatings of the forebody and (2) bond line variations in the aftbody. In the forebody, the thermal history of thermal control coatings may be deduced from the presence of particular semiconducting defect states associated with ZnO, a common emissivity constituent in thermal control coatings. A spatial dependence of this history was mapped for these regions. In the aftbody, luminescing defect states, associated with Si and SiO2 color centers were found along regions of bond variability.

Ultrasensitive turn-on fluorescence detection of Cu2 + based on p-dimethylaminobenzamide derivative and the application to cell imaging

NASA Astrophysics Data System (ADS)

Huang, Peng-Cheng; Fang, Hao; Xiong, Jing-Jing; Wu, Fang-Ying

2017-02-01

A new p-dimethylaminobenzamide derivative based compound BDIH has been synthesized. Cu2 + turned on the fluorescence of compound BDIH with a 1:2 binding stoichiometry. The fluorescent color of compound BDIH shows an evident change from colorless to bright blue upon the addition of Cu2 +, which could be visibly detected by the naked eye under UV light at 365 nm. More importantly, the detection limit was found to be 0.64 nM which is far lower than the maximal allowed concentration of the WHO limit (31.5 μM) for drinking water. This selective ;turn-on; fluorescence sensor was used to identify Cu2 + in living cells using confocal fluorescence microscopy, indicating that compound BDIH has a potential application for selective detection of Cu2 + in organism.

A six-color four-laser mobile platform for multi-spectral fluorescence imaging endoscopy

NASA Astrophysics Data System (ADS)

Black, John F.; Tate, Tyler; Keenan, Molly; Swan, Elizabeth; Utzinger, Urs; Barton, Jennifer

2015-03-01

The properties of multi-spectral fluorescence imaging using deep-UV-illumination have recently been explored using a fiber-coupled thermal source at 280 nm. The resulting images show a remarkable level of contrast thought to result from the signal being overwhelmingly generated in the uppermost few cell layers of tissue, making this approach valuable for the study of diseases that originate in the endothelial tissues of the body. With a view to extending the technique with new wavelengths, and improving beam quality for efficient small core fiber coupling we have developed a mobile self-contained tunable solid-state laser source of deep UV light. An alexandrite laser, lasing at around 750 nm is frequency doubled to produce 375 nm and then tripled to produce 250 nm light. An optical deck added to the system allows other laser sources to be incorporated into the UV beam-line and a lens system has been designed to couple these sources into a single delivery fiber with core diameters down to 50 microns. Our system incorporates five wavelengths [250 nm, 375 nm, 442 nm (HeCd), 543 nm (HeNe) and 638 nm (diode laser)] as the illumination source for a small diameter falloposcope designed for the study of the distal Fallopian tube origins of high grade serous ovarian cancer. The tunability of alexandrite offers the potential to generate other wavelengths in the 720-800, 360-400 and 240-265 nm ranges, plus other non-linear optical conversion techniques taking advantage of the high peak powers of the laser.

A litmus-type colorimetric and fluorometric volatile organic compound sensor based on inkjet-printed polydiacetylenes on paper substrates.

PubMed

Yoon, Bora; Park, In Sung; Shin, Hyora; Park, Hye Jin; Lee, Chan Woo; Kim, Jong-Man

2013-05-14

Inkjet-printed paper-based volatile organic compound (VOC) sensor strips imaged with polydiacetylenes (PDAs) are developed. A microemulsion ink containing bisurethane-substituted diacetylene (DA) monomers, 4BCMU, was inkjet printed onto paper using a conventional inkjet office printer. UV irradiation of the printed image allowed fabrication of blue-colored poly-4BCMU on the paper and the polymer was found to display colorimetric responses to VOCs. Interestingly, a blue-to-yellow color change was observed when the strip was exposed to chloroform vapor, which was accompanied by the generation of green fluorescence. The principal component analysis plot of the color and fluorescence images of the VOC-exposed polymers allowed a more precise discrimination of VOC vapors. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ultrafast Pulse Sequencing for Fast Projective Measurements of Atomic Hyperfine Qubits

NASA Astrophysics Data System (ADS)

Ip, Michael; Ransford, Anthony; Campbell, Wesley

2015-05-01

Projective readout of quantum information stored in atomic hyperfine structure typically uses state-dependent CW laser-induced fluorescence. This method requires an often sophisticated imaging system to spatially filter out the background CW laser light. We present an alternative approach that instead uses simple pulse sequences from a mode-locked laser to affect the same state-dependent excitations in less than 1 ns. The resulting atomic fluorescence occurs in the dark, allowing the placement of non-imaging detectors right next to the atom to improve the qubit state detection efficiency and speed. We also discuss methods of Doppler cooling with mode-locked lasers for trapped ions, where the creation of the necessary UV light is often difficult with CW lasers.

UV Catalysis, Cyanotype Photography, and Sunscreens

NASA Astrophysics Data System (ADS)

Lawrence, Glen D.; Fishelson, Stuart

1999-09-01

This laboratory experiment is intended for a chemistry course for non-science majors. The experiment utilizes one of the earliest photographic processes, the cyanotype process, to demonstrate UV catalysis of chemical reactions. In addition to making photographic prints from negatives, the process can be used to test the effectiveness of sunscreens and the relative efficacy of the SPF (sun protection factor) rating of sunscreens. This is an inexpensive process, requiring solutions of ammonium ferric citrate and potassium ferricyanide, with options to use hydrogen peroxide and ammonium hydroxide solutions. Students can prepare their own UV-sensitized paper with the indicated chemicals and watch the photographic image appear as it is exposed to sunlight or fluorescent UV lamps in a light box designed for use in this experiment. The laboratory experiment should stimulate discussion of UV catalysis, photographic processes and photochemistry, sunscreens, and UV damage to biological organisms. The chemicals used are relatively nontoxic, and the procedure is simple enough to be used by groups of diverse ages and abilities.

Material and Optical Properties of Fluorescent Carbon Quantum Dots Fabricated from Lemon Juice via Hydrothermal Reaction

NASA Astrophysics Data System (ADS)

He, Meiqin; Zhang, Jin; Wang, Hai; Kong, Yanrong; Xiao, Yiming; Xu, Wen

2018-06-01

The water-soluble fluorescent carbon quantum dots (CQDs) are synthesized by utilizing lemon juice as carbon resource via a simple hydrothermal reaction. The obtained CQDs are with an average size of 3.1 nm. They reveal uniform morphology and well-crystalline and can generate bright blue-green light emission under UV or blue light irradiation. We find that the fluorescence from these CQDs is mainly induced by the presence of oxygen-containing groups on the surface and edge of the CQDs. Moreover, we demonstrate that the as-prepared CQDs can be applied to imaging plant cells. This study is related to the fabrication, investigation, and application of newly developed carbon nanostructures.

Plume characteristics and dynamics of UV and IR laser-desorbed oligonucleotides.

PubMed

Merrigan, Tony L; Timson, David J; Hunniford, C Adam; Catney, Martin; McCullough, Robert W

2012-05-01

Laser desorption of dye-tagged oligonucleotides was studied using laser-induced fluorescence imaging. Desorption with ultra violet (UV) and infra-red (IR) lasers resulted in forward directed plumes of molecules. In the case of UV desorption, the initial shot desorbed approximately seven-fold more material than subsequent shots. In contrast, the initial shot in IR desorption resulted in the ejection of less material compared to subsequent shots and these plumes had a component directed along the path of the laser. Thermal equilibrium of the molecules in the plume was achieved after approximately 25 μs with a spread in molecular temperature which was described by a modified Maxwell-Boltzmann equation. Copyright © 2012 Elsevier B.V. All rights reserved.

Laser synthesis and spectroscopy of acetonitrile/silver nanoparticles

NASA Astrophysics Data System (ADS)

Akin, S. T.; Liu, X.; Duncan, M. A.

2015-11-01

Silver nanoparticles with acetonitrile ligands are produced in a laser ablation flow reactor. Excimer laser ablation produces gas phase metal clusters which are thermalized with helium or argon collisions in the flowtube, and reactions with acetonitrile vapor coordinate this ligand to the particle surface. The gaseous mixture is captured in a cryogenic trap; warming produces a solution of excess ligand and coated particles. TEM images reveal particle sizes of 10-30 nm diameter. UV-vis absorption and fluorescence spectra are compared to those of standard silver nanoparticles with surfactant coatings. Deep-UV ligand absorption is strongly enhanced by nanoparticle adsorption.

In situ measurement of VUV/UV radiation from low-pressure microwave-produced plasma in Ar/O2 gas mixtures

NASA Astrophysics Data System (ADS)

Iglesias, E. J.; Mitschker, F.; Fiebrandt, M.; Bibinov, N.; Awakowicz, P.

2017-08-01

Ultraviolet (UV) and vacuum ultraviolet (VUV) spectral irradiance is determined in low-pressure microwave-produced plasma, which is regularly used for polymer surface treatment. The re-emitted fluorescence in the UV/VIS spectral range from a sodium salicylate layer is measured. This fluorescence is related to VUV/UV radiation in different spectral bands based on cut-off filters. The background produced by direct emitted radiation in the fluorescence spectral region is quantified using a specific background filter, thus enabling the use of the whole fluorescence spectral range. A novel procedure is applied to determine the absolute value of the VUV/UV irradiance on a substrate. For that, an independent measurement of the absolute spectral emissivity of the plasma in the UV is performed. The measured irradiances on a substrate from a 25 Pa Ar/O2-produced plasma are in the range of 1015-1016 (photon~ s-1 cm-2). These values include the contribution from impurities present in the discharge.

Differential laser-induced perturbation spectroscopy and fluorescence imaging for biological and materials sensing

NASA Astrophysics Data System (ADS)

Burton, Dallas Jonathan

The field of laser-based diagnostics has been a topic of research in various fields, more specifically for applications in environmental studies, military defense technologies, and medicine, among many others. In this dissertation, a novel laser-based optical diagnostic method, differential laser-induced perturbation spectroscopy (DLIPS), has been implemented in a spectroscopy mode and expanded into an imaging mode in combination with fluorescence techniques. The DLIPS method takes advantage of deep ultraviolet (UV) laser perturbation at sub-ablative energy fluences to photochemically cleave bonds and alter fluorescence signal response before and after perturbation. The resulting difference spectrum or differential image adds more information about the target specimen, and can be used in combination with traditional fluorescence techniques for detection of certain materials, characterization of many materials and biological specimen, and diagnosis of various human skin conditions. The differential aspect allows for mitigation of patient or sample variation, and has the potential to develop into a powerful, noninvasive optical sensing tool. The studies in this dissertation encompass efforts to continue the fundamental research on DLIPS including expansion of the method to an imaging mode. Five primary studies have been carried out and presented. These include the use of DLIPS in a spectroscopy mode for analysis of nitrogen-based explosives on various substrates, classification of Caribbean fruit flies versus Caribbean fruit flies that have been irradiated with gamma rays, and diagnosis of human skin cancer lesions. The nitrogen-based explosives and Caribbean fruit flies have been analyzed with the DLIPS scheme using the imaging modality, providing complementary information to the spectroscopic scheme. In each study, a comparison between absolute fluorescence signals and DLIPS responses showed that DLIPS statistically outperformed traditional fluorescence techniques with regards to regression error and classification.

NASA Tech Briefs, April 2004

NASA Technical Reports Server (NTRS)

2004-01-01

Topics covered include: Analysis of SSEM Sensor Data Using BEAM; Hairlike Percutaneous Photochemical Sensors; Video Guidance Sensors Using Remotely Activated Targets; Simulating Remote Sensing Systems; EHW Approach to Temperature Compensation of Electronics; Polymorphic Electronic Circuits; Micro-Tubular Fuel Cells; Whispering-Gallery-Mode Tunable Narrow-Band-Pass Filter; PVM Wrapper; Simulation of Hyperspectral Images; Algorithm for Controlling a Centrifugal Compressor; Hybrid Inflatable Pressure Vessel; Double-Acting, Locking Carabiners; Position Sensor Integral with a Linear Actuator; Improved Electromagnetic Brake; Flow Straightener for a Rotating-Drum Liquid Separator; Sensory-Feedback Exoskeletal Arm Controller; Active Suppression of Instabilities in Engine Combustors; Fabrication of Robust, Flat, Thinned, UV-Imaging CCDs; Chemical Thinning Process for Fabricating UV-Imaging CCDs; Pseudoslit Spectrometer; Waste-Heat-Driven Cooling Using Complex Compound Sorbents; Improved Refractometer for Measuring Temperatures of Drops; Semiconductor Lasers Containing Quantum Wells in Junctions; Phytoplankton-Fluorescence-Lifetime Vertical Profiler; Hexagonal Pixels and Indexing Scheme for Binary Images; Finding Minimum-Power Broadcast Trees for Wireless Networks; and Automation of Design Engineering Processes.

Development of a Bioaerosol single particle detector (BIO IN) for the Fast Ice Nucleus CHamber FINCH

NASA Astrophysics Data System (ADS)

Bundke, U.; Reimann, B.; Nillius, B.; Jaenicke, R.; Bingemer, H.

2010-02-01

In this work we present the setup and first tests of our new BIO IN detector. This detector was constructed to classify atmospheric ice nuclei (IN) for their biological content. It is designed to be coupled to the Fast Ice Nucleus CHamber FINCH. If one particle acts as an ice nucleus, it will be at least partly covered with ice at the end of the development section of the FINCH chamber. The device combines an auto-fluorescence detector and a circular depolarization detector for simultaneous detection of biological material and discrimination between water droplets, ice crystals and non activated large aerosol particles. The excitation of biological material with UV light and analysis of auto-fluorescence is a common principle used for flow cytometry, fluorescence microscopy, spectroscopy and imaging. The detection of auto-fluorescence of airborne single particles demands some more experimental effort. However, expensive commercial sensors are available for special purposes, e.g. size distribution measurements. But these sensors will not fit the specifications needed for the FINCH IN counter (e.g. high sample flow of up 10 LPM). The newly developed -low cost- BIO IN sensor uses a single high-power UV LED for the electronic excitation instead of much more expensive UV lasers. Other key advantages of the new sensor are the low weight, compact size, and the little effect on the aerosol sample, which allows it to be coupled with other instruments for further analysis. The instrument will be flown on one of the first missions of the new German research aircraft "HALO" (High Altitude and LOng range).

Fluorometric Biosniffer Camera "Sniff-Cam" for Direct Imaging of Gaseous Ethanol in Breath and Transdermal Vapor.

PubMed

Arakawa, Takahiro; Sato, Toshiyuki; Iitani, Kenta; Toma, Koji; Mitsubayashi, Kohji

2017-04-18

Various volatile organic compounds can be found in human transpiration, breath and body odor. In this paper, a novel two-dimensional fluorometric imaging system, known as a "sniffer-cam" for ethanol vapor released from human breath and palm skin was constructed and validated. This imaging system measures ethanol vapor concentrations as intensities of fluorescence through an enzymatic reaction induced by alcohol dehydrogenase (ADH). The imaging system consisted of multiple ultraviolet light emitting diode (UV-LED) excitation sheet, an ADH enzyme immobilized mesh substrate and a high-sensitive CCD camera. This imaging system uses ADH for recognition of ethanol vapor. It measures ethanol vapor by measuring fluorescence of nicotinamide adenine dinucleotide (NADH), which is produced by an enzymatic reaction on the mesh. This NADH fluorometric imaging system achieved the two-dimensional real-time imaging of ethanol vapor distribution (0.5-200 ppm). The system showed rapid and accurate responses and a visible measurement, which could lead to an analysis of metabolism function at real time in the near future.

Plasmonic enhancement of ultraviolet fluorescence

NASA Astrophysics Data System (ADS)

Jiao, Xiaojin

Plasmonics relates to the interaction between electromagnetic radiation and conduction electrons at metallic interfaces or in metallic nanostructures. Surface plasmons are collective electron oscillations at a metal surface, which can be manipulated by shape, texture and material composition. Plasmonic applications cover a broad spectrum from visible to near infrared, including biosensing, nanolithography, spectroscopy, optoelectronics, photovoltaics and so on. However, there remains a gap in this activity in the ultraviolet (UV, < 400 nm), where significant opportunity exists for both fundamental and application research. Motivating factors in the study of UV Plasmonics are the direct access to biomolecular resonances and native fluorescence, resonant Raman scattering interactions, and the potential for exerting control over photochemical reactions. This dissertation aims to fill in the gap of Plasmonics in the UV with efforts of design, fabrication and characterization of aluminium (Al) and magnesium (Mg) nanostructures for the application of label-free bimolecular detection via native UV fluorescence. The first contribution of this dissertation addresses the design of Al nanostructures in the context of UV fluorescence enhancement. A design method that combines analytical analysis with numerical simulation has been developed. Performance of three canonical plasmonic structures---the dipole antenna, bullseye nanoaperture and nanoaperture array---has been compared. The optimal geometrical parameters have been determined. A novel design of a compound bullseye structure has been proposed and numerically analyzed for the purpose of compensating for the large Stokes shift typical of UV fluorescence. Second, UV lifetime modification of diffusing molecules by Al nanoapertures has been experimentally demonstrated for the first time. Lifetime reductions of ~3.5x have been observed for the high quantum yield (QY) laser dye p-terphenyl in a 60 nm diameter aperture with 50 nm undercut. Furthermore, quantum-yield-dependence of lifetime reduction has been experimentally demonstrated for the first time. Lifetime reduction as a function of aperture size and native quantum yield has been accurately predicted by simulation. Simulation further predicts greater net fluorescence enhancement for tryptophan compared to p-terphenyl. In order to increase fluorescence enhancement, the "poor" molecules and structures with proper undercuts are required. Third, UV lifetime modification by Mg nanoapertures has been experimentally demonstrated for the fisrt time. Lifetime reductions of ~13x have been observed for the laser dye p-terphenyl with high QY in a 50 nm diameter aperture with 125 nm undercut. In addition, extraordinary optical transmission of Mg nanohole arrays in the UV has been measured for the first time. By using Al as a reference, the feasibility of applying Mg in the UV plasmonic applications has been evaluated both numerically and experimentally. Finally, this work has established a methodology for the study of plasmonic enhancement of UV fluorescence, including design method, thin-film characterization, nanofabrication with focus ion beam milling, and fluorescence measurement. It has paved the way for more extensive research on UV fluorescence enhancement.

Aminoquinoline based highly sensitive fluorescent sensor for lead(II) and aluminum(III) and its application in live cell imaging.

PubMed

Anand, Thangaraj; Sivaraman, Gandhi; Mahesh, Ayyavu; Chellappa, Duraisamy

2015-01-01

We have synthesized a new probe 5-((anthracen-9-ylmethylene) amino)quinolin-10-ol (ANQ) based on anthracene platform. The probe was tested for its sensing behavior toward heavy metal ions Hg(2+), Pb(2+), light metal Al(3+) ion, alkali, alkaline earth, and transition metal ions by UV-visible and fluorescent techniques in ACN/H2O mixture buffered with HEPES (pH 7.4). It shows high selectivity toward sensing Pb(2+)/Al(3+) metal ions. Importantly, 10-fold and 5- fold fluorescence enhancement at 429 nm was observed for probe upon complexation with Pb(2+) and Al(3+) ions, respectively. This fluorescence enhancement is attributable to the prevention of photoinduced electron transfer. The photonic studies indicate that the probe can be adopted as a sensitive fluorescent chemosensor for Pb(2+) and Al(3+) ions. Copyright © 2014 Elsevier B.V. All rights reserved.

Evaluation of optimal DNA staining for triggering by scanning fluorescence microscopy (SFM)

NASA Astrophysics Data System (ADS)

Mittag, Anja; Marecka, Monika; Pierzchalski, Arkadiusz; Malkusch, Wolf; Bocsi, József; Tárnok, Attila

2009-02-01

In imaging and flow cytometry, DNA staining is a common trigger signal for cell identification. Selection of the proper DNA dye is restricted by the hardware configuration of the instrument. The Zeiss Imaging Solutions GmbH (München, Germany) introduced a new automated scanning fluorescence microscope - SFM (Axio Imager.Z1) which combines fluorescence imaging with cytometric parameters measurement. The aim of the study was to select optimal DNA dyes as trigger signal in leukocyte detection and subsequent cytometric analysis of double-labeled leukocytes by SFM. Seven DNA dyes (DAPI, Hoechst 33258, Hoechst 33342, POPO-3, PI, 7-AAD, and TOPRO-3) were tested and found to be suitable for the implemented filtersets (fs) of the SFM (fs: 49, fs: 44, fs: 20). EDTA blood was stained after erythrocyte lysis with DNA dye. Cells were transferred on microscopic slides and embedded in fluorescent mounting medium. Quality of DNA fluorescence signal as well as spillover signals were analyzed by SFM. CD45-APC and CD3-PE as well as CD4-FITC and CD8-APC were selected for immunophenotyping and used in combination with Hoechst. Within the tested DNA dyes DAPI showed relatively low spillover and the best CV value. Due to the low spillover of UV DNA dyes a triple staining of Hoechst and APC and PE (or APC and FITC, respectively) could be analyzed without difficulty. These results were confirmed by FCM measurements. DNA fluorescence is applicable for identifying and triggering leukocytes in SFM analyses. Although some DNA dyes exhibit strong spillover in other fluorescence channels, it was possible to immunophenotype leukocytes. DAPI seems to be best suitable for use in the SFM system and will be used in protocol setups as primary parameter.

Onychomycosis diagnosis using fluorescence and infrared imaging systems

NASA Astrophysics Data System (ADS)

da Silva, Ana Paula; Fortunato, Thereza Cury; Stringasci, Mirian D.; Kurachi, Cristina; Bagnato, Vanderlei S.; Inada, Natalia M.

2015-06-01

Onychomycosis is a common disease of the nail plate, constituting approximately half of all cases of nail infection. Onychomycosis diagnosis is challenging because it is hard to distinguish from other diseases of the nail lamina such as psoriasis, lichen ruber or eczematous nails. The existing methods of diagnostics so far consist of clinical and laboratory analysis, such as: Direct Mycological examination and culture, PCR and histopathology with PAS staining. However, they all share certain disadvantages in terms of sensitivity and specificity, time delay, or cost. This study aimed to evaluate the use of infrared and fluorescence imaging as new non-invasive diagnostic tools in patients with suspected onychomycosis, and compare them with established techniques. For fluorescence analysis, a Clinical Evince (MM Optics®) was used, which consists of an optical assembly with UV LED light source wavelength 400 nm +/- 10 nm and the maximum light intensity: 40 mW/cm2 +/- 20%. For infrared analysis, a Fluke® Camera FKL model Ti400 was used. Patients with onychomycosis and control group were analyzed for comparison. The fluorescence images were processed using MATLAB® routines, and infrared images were analyzed using the SmartView® 3.6 software analysis provided by the company Fluke®. The results demonstrated that both infrared and fluorescence could be complementary to diagnose different types of onychomycosis lesions. The simplicity of operation, quick response and non-invasive assessment of the nail patients in real time, are important factors to be consider for an implementation.

Versatile Polymer Nanoparticles as Two-Photon-Triggered Photosensitizers for Simultaneous Cellular, Deep-Tissue Imaging, and Photodynamic Therapy.

PubMed

Guo, Liang; Ge, Jiechao; Liu, Qian; Jia, Qingyan; Zhang, Hongyan; Liu, Weimin; Niu, Guangle; Liu, Sha; Gong, Jianru; Hackbarth, Steffen; Wang, Pengfei

2017-06-01

Clinical applications of current photodynamic therapy (PDT) photosensitizers (PSs) are often limited by their absorption in the UV-vis range that possesses limited tissue penetration ability, leading to ineffective therapeutic response for deep-seated tumors. Alternatively, two-photon excited PS (TPE-PS) using NIR light triggered is one the most promising candidates for PDT improvement. Herein, multimodal polymer nanoparticles (PNPs) from polythiophene derivative as two-photon fluorescence imaging as well as two-photon-excited PDT agent are developed. The prepared PNPs exhibit excellent water dispersibility, high photostability and pH stability, strong fluorescence brightness, and low dark toxicity. More importantly, the PNPs also possess other outstanding features including: (1) the high 1 O 2 quantum yield; (2) the strong two-photon-induced fluorescence and efficient 1 O 2 generation; (3) the specific accumulation in lysosomes of HeLa cells; and (4) the imaging detection depth up to 2100 µm in the mock tissue under two-photon. The multifunctional PNPs are promising candidates as TPE-PDT agent for simultaneous cellular, deep-tissue imaging, and highly efficient in vivo PDT of cancer. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescent porous silicon biological probes with high quantum efficiency and stability.

PubMed

Tu, Chang-Ching; Chou, Ying-Nien; Hung, Hsiang-Chieh; Wu, Jingda; Jiang, Shaoyi; Lin, Lih Y

2014-12-01

We demonstrate porous silicon biological probes as a stable and non-toxic alternative to organic dyes or cadmium-containing quantum dots for imaging and sensing applications. The fluorescent silicon quantum dots which are embedded on the porous silicon surface are passivated with carboxyl-terminated ligands through stable Si-C covalent bonds. The porous silicon bio-probes have shown photoluminescence quantum yield around 50% under near-UV excitation, with high photochemical and thermal stability. The bio-probes can be efficiently conjugated with antibodies, which is confirmed by a standard enzyme-linked immunosorbent assay (ELISA) method.

Dynamin-Related Protein 1 Translocates from the Cytosol to Mitochondria during UV-Induced Apoptosis

NASA Astrophysics Data System (ADS)

Zhang, Zhenzhen; Wu, Shengnan; Feng, Jie

2011-01-01

Mitochondria are dynamic structures that frequently divide and fuse with one another to form interconnecting network. This network disintegrates into punctiform organelles during apoptosis. However, the mechanisms involved in these processes are still not well characterized. In this study, we investigate the role of dynamin-related protein 1 (Drp1), a large GTPase that mediates outer mitochondrial membrane fission, in mitochondrial dynamics in response to UV irradiation in human lung adenocarcinoma cells (ASTC-α-1) and HeLa cells. Using time-lapse fluorescent imaging, we find that Drp1 primarily distributes in cytosol under physiological conditions. After UV treatment, Drp1 translocates from cytosol to mitochondria, indicating the enhancement of Drp1 mitochondrial accumulation. Our results suggest that Drp1 is involved in the regulation of transition from an interconnecting network to a punctiform mitochondrial phenotype during UV-induced apoptosis.

Calibration and validation of projection lithography in chemically amplified resist systems using fluorescence imaging

NASA Astrophysics Data System (ADS)

Mason, Michael D.; Ray, Krishanu; Feke, Gilbert D.; Grober, Robert D.; Pohlers, Gerd; Cameron, James F.

2003-05-01

Coumarin 6 (C6), a pH sensitive fluorescent molecule were doped into commercial resist systems to demonstrate a cost-effective fluorescence microscopy technique for detecting latent photoacid images in exposed chemically amplified resist films. The fluorescenec image contrast is optimized by carefully selecting optical filters to match the spectroscopic properties of C6 in the resist matrices. We demonstrate the potential of this technique for two sepcific non-invasive applications. First, a fast, conventient, fluorescence technique is demonstrated for determination of quantum yeidsl of photo-acid generation. Since the Ka of C6 in the 193nm resist system lies wihtin the range of acid concentrations that can be photogenerated, we have used this technique to evaluate the acid generation efficiency of various photo-acid generators (PAGs). The technique is based on doping the resist formulations containing the candidate PAGs with C6, coating one wafer per PAG, patterning the wafer with a dose ramp and spectroscopically imaging the wafers. The fluorescence of each pattern in the dose ramp is measured as a single image and analyzed with the optical titration model. Second, a nondestructive in-line diagnostic technique is developed for the focus calibration and validation of a projection lithography system. Our experimental results show excellent correlation between the fluorescence images and scanning electron microscope analysis of developed features. This technique has successfully been applied in both deep UV resists e.g., Shipley UVIIHS resist and 193 nm resists e.g., Shipley Vema-type resist. This method of focus calibration has also been extended to samples with feature sizes below the diffraction limit where the pitch between adjacent features is on the order of 300 nm. Image capture, data analysis, and focus latitude verification are all computer controlled from a single hardware/software platform. Typical focus calibration curves can be obtained within several minutes.

Picturing pathogen infection in plants.

PubMed

Barón, Matilde; Pineda, Mónica; Pérez-Bueno, María Luisa

2016-09-01

Several imaging techniques have provided valuable tools to evaluate the impact of biotic stress on host plants. The use of these techniques enables the study of plant-pathogen interactions by analysing the spatial and temporal heterogeneity of foliar metabolism during pathogenesis. In this work we review the use of imaging techniques based on chlorophyll fluorescence, multicolour fluorescence and thermography for the study of virus, bacteria and fungi-infected plants. These studies have revealed the impact of pathogen challenge on photosynthetic performance, secondary metabolism, as well as leaf transpiration as a promising tool for field and greenhouse management of diseases. Images of standard chlorophyll fluorescence (Chl-F) parameters obtained during Chl-F induction kinetics related to photochemical processes and those involved in energy dissipation, could be good stress indicators to monitor pathogenesis. Changes on UV-induced blue (F440) and green fluorescence (F520) measured by multicolour fluorescence imaging in pathogen-challenged plants seem to be related with the up-regulation of the plant secondary metabolism and with an increase in phenolic compounds involved in plant defence, such as scopoletin, chlorogenic or ferulic acids. Thermal imaging visualizes the leaf transpiration map during pathogenesis and emphasizes the key role of stomata on innate plant immunity. Using several imaging techniques in parallel could allow obtaining disease signatures for a specific pathogen. These techniques have also turned out to be very useful for presymptomatic pathogen detection, and powerful non-destructive tools for precision agriculture. Their applicability at lab-scale, in the field by remote sensing, and in high-throughput plant phenotyping, makes them particularly useful. Thermal sensors are widely used in crop fields to detect early changes in leaf transpiration induced by both air-borne and soil-borne pathogens. The limitations of measuring photosynthesis by Chl-F at the canopy level are being solved, while the use of multispectral fluorescence imaging is very challenging due to the type of light excitation that is used.

Detecting peanuts inoculated with toxigenic and atoxienic Aspergillus flavus strains with fluorescence hyperspectral imagery

NASA Astrophysics Data System (ADS)

Xing, Fuguo; Yao, Haibo; Hruska, Zuzana; Kincaid, Russell; Zhu, Fengle; Brown, Robert L.; Bhatnagar, Deepak; Liu, Yang

2017-05-01

Aflatoxin contamination in peanut products has been an important and long-standing problem around the world. Produced mainly by Aspergillus flavus and Aspergillus parasiticus, aflatoxins are the most toxic and carcinogenic compounds among toxins. This study investigated the application of fluorescence visible near-infrared (VNIR) hyperspectral images to assess the spectral difference between peanut kernels inoculated with toxigenic and atoxigenic inocula of A. flavus and healthy kernels. Peanut kernels were inoculated with NRRL3357, a toxigenic strain of A. flavus, and AF36, an atoxigenic strain of A. flavus, respectively. Fluorescence hyperspectral images under ultraviolet (UV) excitation were recorded on peanut kernels with and without skin. Contaminated kernels exhibited different fluorescence features compared with healthy kernels. For the kernels without skin, the inoculated kernels had a fluorescence peaks shifted to longer wavelengths with lower intensity than healthy kernels. In addition, the fluorescence intensity of peanuts without skin was higher than that of peanuts with skin (10 times). The fluorescence spectra of kernels with skin are significantly different from that of the control group (p<0.001). Furthermore, the fluorescence intensity of the toxigenic, AF3357 peanuts with skin was lower than that of the atoxigenic AF36 group. Discriminate analysis showed that the inoculation group can be separated from the controls with 100% accuracy. However, the two inoculation groups (AF3357 vis AF36) can be separated with only ∼80% accuracy. This study demonstrated the potential of fluorescence hyperspectral imaging techniques for screening of peanut kernels contaminated with A. flavus, which could potentially lead to the production of rapid and non-destructive scanning-based detection technology for the peanut industry.

Portable hyperspectral imager with continuous wave green laser for identification and detection of untreated latent fingerprints on walls.

PubMed

Nakamura, Atsushi; Okuda, Hidekazu; Nagaoka, Takashi; Akiba, Norimitsu; Kurosawa, Kenji; Kuroki, Kenro; Ichikawa, Fumihiko; Torao, Akira; Sota, Takayuki

2015-09-01

Untreated latent fingerprints are known to exhibit fluorescence under UV laser excitation. Previously, the hyperspectral imager (HSI) has been primarily evaluated in terms of its potential to enhance the sensitivity of latent fingerprint detection following treatment by conventional chemical methods in the forensic science field. In this study however, the potential usability of the HSI for the visualization and detection of untreated latent fingerprints by measuring their inherent fluorescence under continuous wave (CW) visible laser excitation was examined. Its potential to undertake spectral separation of overlapped fingerprints was also evaluated. The excitation wavelength dependence of fluorescent images was examined using an untreated palm print on a steel based wall, and it was found that green laser excitation is superior to blue and yellow lasers' excitation for the production of high contrast fluorescence images. In addition, a spectral separation method for overlapped fingerprints/palm prints on a plaster wall was proposed using new images converted by the division and subtraction of two single wavelength images constructed based on measured hyperspectral data (HSD). In practical tests, the relative isolation of two overlapped fingerprints/palm prints was successful in twelve out of seventeen cases. Only one fingerprint/palm print was extracted for an additional three cases. These results revealed that the feasibility of overlapped fingerprint/palm print spectral separation depends on the difference in the temporal degeneration of each fluorescence spectrum. The present results demonstrate that a combination of a portable HSI and CW green laser has considerable potential for the identification and detection of untreated latent fingerprints/palm prints on the walls under study, while the use of HSD makes it practically possible for doubly overlapped fingerprints/palm prints to be separated spectrally. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Mechanism of UV-Induced Damage to Mammalian Collagen

DTIC Science & Technology

2014-12-12

enough to use under physiological conditions. We carried out UV photolysis with a UVG – 11 short wavelength hand lamp that emits primarily254 nm. We have...photochemical interaction between UV and mammalian collagen as functions of excitation wavelengths, temperature, fluorescence spectral distribution, and...but has little fluorescence at 325/400 nm (dityrosine). The 325/400 band INCREASES with UV - 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND SUBTITLE 13

A Green Fluorescent Protein with Photoswitchable Emission from the Deep Sea

PubMed Central

Vogt, Alexander; D'Angelo, Cecilia; Oswald, Franz; Denzel, Andrea; Mazel, Charles H.; Matz, Mikhail V.; Ivanchenko, Sergey; Nienhaus, G. Ulrich; Wiedenmann, Jörg

2008-01-01

A colorful variety of fluorescent proteins (FPs) from marine invertebrates are utilized as genetically encoded markers for live cell imaging. The increased demand for advanced imaging techniques drives a continuous search for FPs with new and improved properties. Many useful FPs have been isolated from species adapted to sun-flooded habitats such as tropical coral reefs. It has yet remained unknown if species expressing green fluorescent protein (GFP)-like proteins also exist in the darkness of the deep sea. Using a submarine-based and -operated fluorescence detection system in the Gulf of Mexico, we discovered ceriantharians emitting bright green fluorescence in depths between 500 and 600 m and identified a GFP, named cerFP505, with bright fluorescence emission peaking at 505 nm. Spectroscopic studies showed that ∼15% of the protein bulk feature reversible ON/OFF photoswitching that can be induced by alternating irradiation with blue und near-UV light. Despite being derived from an animal adapted to essentially complete darkness and low temperatures, cerFP505 maturation in living mammalian cells at 37°C, its brightness and photostability are comparable to those of EGFP and cmFP512 from shallow water species. Therefore, our findings disclose the deep sea as a potential source of GFP-like molecular marker proteins. PMID:19018285

Classification of corn kernels contaminated with aflatoxins using fluorescence and reflectance hyperspectral images analysis

NASA Astrophysics Data System (ADS)

Zhu, Fengle; Yao, Haibo; Hruska, Zuzana; Kincaid, Russell; Brown, Robert; Bhatnagar, Deepak; Cleveland, Thomas

2015-05-01

Aflatoxins are secondary metabolites produced by certain fungal species of the Aspergillus genus. Aflatoxin contamination remains a problem in agricultural products due to its toxic and carcinogenic properties. Conventional chemical methods for aflatoxin detection are time-consuming and destructive. This study employed fluorescence and reflectance visible near-infrared (VNIR) hyperspectral images to classify aflatoxin contaminated corn kernels rapidly and non-destructively. Corn ears were artificially inoculated in the field with toxigenic A. flavus spores at the early dough stage of kernel development. After harvest, a total of 300 kernels were collected from the inoculated ears. Fluorescence hyperspectral imagery with UV excitation and reflectance hyperspectral imagery with halogen illumination were acquired on both endosperm and germ sides of kernels. All kernels were then subjected to chemical analysis individually to determine aflatoxin concentrations. A region of interest (ROI) was created for each kernel to extract averaged spectra. Compared with healthy kernels, fluorescence spectral peaks for contaminated kernels shifted to longer wavelengths with lower intensity, and reflectance values for contaminated kernels were lower with a different spectral shape in 700-800 nm region. Principal component analysis was applied for data compression before classifying kernels into contaminated and healthy based on a 20 ppb threshold utilizing the K-nearest neighbors algorithm. The best overall accuracy achieved was 92.67% for germ side in the fluorescence data analysis. The germ side generally performed better than endosperm side. Fluorescence and reflectance image data achieved similar accuracy.

Masked red-emitting carbopyronine dyes with photosensitive 2-diazo-1-indanone caging group.

PubMed

Kolmakov, Kirill; Wurm, Christian; Sednev, Maksim V; Bossi, Mariano L; Belov, Vladimir N; Hell, Stefan W

2012-03-01

Caged near-IR emitting fluorescent dyes are in high demand in optical microscopy but up to now were unavailable. We discovered that the combination of a carbopyronine dye core and a photosensitive 2-diazo-1-indanone residue leads to masked near-IR emitting fluorescent dyes. Illumination of these caged dyes with either UV or visible light (λ < 420 nm) efficiently generates fluorescent compounds with absorption and emission at 635 nm and 660 nm, respectively. A high-yielding synthetic route with attractive possibilities for further dye design is described in detail. Good photostability, high contrast, and a large fluorescence quantum yield after uncaging are the most important features of the new compounds for non-invasive imaging in high-resolution optical microscopy. For use in immunolabelling the caged dyes were decorated with a (hydrophilic) linker and an (activated) carboxyl group.

Hyperspectral imaging technique for detection of poultry fecal residues on food processing equipments

NASA Astrophysics Data System (ADS)

Cho, Byoung-Kwan; Kim, Moon S.; Chen, Yud-Ren

2005-11-01

Emerging concerns about safety and security in current mass production of food products necessitate rapid and reliable inspection for contaminant-free products. Diluted fecal residues on poultry processing plant equipment surface, not easily discernable from water by human eye, are contamination sources for poultry carcasses. Development of sensitive detection methods for fecal residues is essential to ensure safe production of poultry carcasses. Hyperspectral imaging techniques have shown good potential for detecting of the presence of fecal and other biological substances on food and processing equipment surfaces. In this study, use of high spatial resolution hyperspectral reflectance and fluorescence imaging (with UV-A excitation) is presented as a tool for selecting a few multispectral bands to detect diluted fecal and ingesta residues on materials used for manufacturing processing equipment. Reflectance and fluorescence imaging methods were compared for potential detection of a range of diluted fecal residues on the surfaces of processing plant equipment. Results showed that low concentrations of poultry feces and ingesta, diluted up to 1:100 by weight with double distilled water, could be detected using hyperspectral fluorescence images with an accuracy of 97.2%. Spectral bands determined in this study could be used for developing a real-time multispectral inspection device for detection of harmful organic residues on processing plant equipment.

Light transmission and ultraviolet protection of contact lenses under artificial illumination.

PubMed

Artigas, José M; Navea, Amparo; García-Domene, M Carmen; Gené, Andrés; Artigas, Cristina

2016-04-01

To determine the spectral transmission of contact lenses (CLs), with and without an ultraviolet (UV) filter to evaluate their capacity for protection under UV radiation from artificial illumination (incandescent, fluorescent, xenon (Xe) lamps, or white LEDs (light-emitting diode)). The transmission curves of nine soft CLs were obtained by using a PerkinElmer Lambda 35 UV-vis spectrophotometer. A CIE standard was used for the emission spectra of incandescent and fluorescent lamps, and Xe lamps and white LEDs were measured by using an International Light Technologies ILT-950 spectroradiometer. Five of the nine soft CLs analysed state that they incorporate UV filters, but the other four do not specify anything in this regard. The spectral transmission of all the CLs studied is excellent in the visible region. The CLs with UV filters filter out this radiation more or less effectively. Xe lamps emit a part in the UV region. Incandescent, fluorescent and white LEDs do not emit at all in the UV. Incorporating UV filters is important when the illumination is from a Xe lamp since this light source emits in the UV region. This, however, does not occur with incandescent and fluorescent lamps or white LEDs. The CLs that do incorporate UV filters meet all the standard requirements that the U.S. FDA (Food and Drug Administration) has for UV-blocking CLs Class II (OcularScience, CooperVision and Neolens), and AcuvueMoist and HydronActifresh400 even comply with the stricter Class I. The CLs without UV filters let UVA, UVB and even some UVC through. Copyright © 2015. Published by Elsevier Ltd.

Nitrogen-doped carbon dots originating from unripe peach for fluorescent bioimaging and electrocatalytic oxygen reduction reaction.

PubMed

Atchudan, Raji; Edison, Thomas Nesakumar Jebakumar Immanuel; Lee, Yong Rok

2016-11-15

This paper reports the robust hydrothermal synthesis of nitrogen doped carbon dots (N-CDs) using the unripe fruit of Prunus persica (peach) as the carbon precursor and aqueous ammonia as the nitrogen source. The optical properties of synthesized N-CDs were characterized by ultraviolet visible (UV-Vis) and fluorescence spectroscopy techniques. The synthesized N-CDs were emitted blue light when excitated with a portable UV lamp. The materials with the optical properties were characterized further by high resolution transmission electron microscope (HRTEM), X-ray diffraction (XRD), Raman, Fourier transform infrared (FT-IR) and X-ray photoelectron spectroscopy (XPS). The mean size of the N-CDs was approximately 8nm, as calculated from the HRTEM image. The d-spacing of N-CDs, calculated using Bragg law, was approximately 0.21nm, which was consistent with the interlayer distance calculated from the HRTEM image. FT-IR spectroscopy and XPS revealed the presence of the phytoconstituents functionalities of peach fruit over the N-CDs surface and a high level of nitrogen doping on carbon dots (CDs) was confirmed by XPS studies. These results suggest that the unripe fruit extract of peach is an ideal candidate for the preparation of N-CDs. The resulting N-CDs showed excellent optical properties in water. The synthesized N-CDs exhibited a high fluorescence quantum yield and low cytotoxicity, and can be used as fluorescence imaging probes. In addition, the N-CDs were catalytically activite towards the oxygen reduction reaction (ORR). The N-CDs exhibited good catalytic activity in an alkaline medium (0.1M KOH) with a remarkable ORR of approximately 0.72V vs reversible hydrogen electrode (RHE), and O2 reduction follows mainly a 2 electron pathway by being reduced to hydrogen peroxide. The 2-electron reduction pathway is used in industry for H2O2 production. Copyright © 2016 Elsevier Inc. All rights reserved.

Capillary electrophoresis hyphenated with UV-native-laser induced fluorescence detection (CE/UV-native-LIF).

PubMed

Couderc, François; Ong-Meang, Varravaddheay; Poinsot, Véréna

2017-01-01

Native laser-induced fluorescence using UV lasers associated to CE offers now a large related literature, for now 30 years. The main works have been performed using very expensive Ar-ion lasers emitting at 257 and 275 nm. They are not affordable for routine analyses, but have numerous applications such as protein, catecholamine, and indolamine analysis. Some other lasers such as HeCd 325 nm have been used but only for few applications. Diode lasers, emitting at 266 nm, cheaper, are extensively used for the same topics, even if the obtained sensitivity is lower than the one observed using the costly UV-Ar-ion lasers. This review presents various CE or microchips applications and different UV lasers used for the excitation of native fluorescence. We showed that CE/Native UV laser induced fluorescence detection is very sensitive for detection as well as small aromatic biomolecules than proteins containing Trp and Tyr amino acids. Moreover, it is a simple way to analyze biomolecules without derivatization. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Novel use of UV broad-band excitation and stretched exponential function in the analysis of fluorescent dissolved organic matter: study of interaction between protein and humic-like components

NASA Astrophysics Data System (ADS)

Panigrahi, Suraj Kumar; Mishra, Ashok Kumar

2017-09-01

A combination of broad-band UV radiation (UV A and UV B; 250-400 nm) and a stretched exponential function (StrEF) has been utilised in efforts towards convenient and sensitive detection of fluorescent dissolved organic matter (FDOM). This approach enables accessing the gross fluorescence spectral signature of both protein-like and humic-like components in a single measurement. Commercial FDOM components are excited with the broad-band UV excitation; the variation of spectral profile as a function of varying component ratio is analysed. The underlying fluorescence dynamics and non-linear quenching of amino acid moieties are studied with the StrEF (exp(-V[Q] β )). The complex quenching pattern reflects the inner filter effect (IFE) as well as inter-component interactions. The inter-component interactions are essentially captured through the ‘sphere of action’ and ‘dark complex’ models. The broad-band UV excitation ascertains increased excitation energy, resulting in increased population density in the excited state and thereby resulting in enhanced sensitivity.

Practical implementation, characterization and applications of a multi-colour time-gated luminescence microscope.

PubMed

Zhang, Lixin; Zheng, Xianlin; Deng, Wei; Lu, Yiqing; Lechevallier, Severine; Ye, Zhiqiang; Goldys, Ewa M; Dawes, Judith M; Piper, James A; Yuan, Jingli; Verelst, Marc; Jin, Dayong

2014-10-13

Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (Giardia lamblia stained with a red europium probe and Cryptosporidium parvum with a green terbium probe) at UV wavelengths (320-400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y₂O₂S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment.

Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope

NASA Astrophysics Data System (ADS)

Zhang, Lixin; Zheng, Xianlin; Deng, Wei; Lu, Yiqing; Lechevallier, Severine; Ye, Zhiqiang; Goldys, Ewa M.; Dawes, Judith M.; Piper, James A.; Yuan, Jingli; Verelst, Marc; Jin, Dayong

2014-10-01

Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (Giardia lamblia stained with a red europium probe and Cryptosporidium parvum with a green terbium probe) at UV wavelengths (320-400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y2O2S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment.

Eye Disease Resulting From Increased Use of Fluorescent Lighting as a Climate Change Mitigation Strategy

PubMed Central

Walls, Kelvin L.; Benke, Geza

2011-01-01

Increased use of fluorescent lighting as a climate change mitigation strategy may increase eye disease. The safe range of light to avoid exposing the eye to potentially damaging ultraviolet (UV) radiation is 2000 to 3500K and greater than 500 nanometers. Some fluorescent lights fall outside this safe range. Fluorescent lighting may increase UV-related eye diseases by up to 12% and, according to our calculations, may cause an additional 3000 cases of cataracts and 7500 cases of pterygia annually in Australia. Greater control of UV exposure from fluorescent lights is required. This may be of particular concern for aging populations in developed countries and countries in northern latitudes where there is a greater dependence on artificial lighting. PMID:22021286

Toward intravascular morphological and biochemical imaging of atherosclerosis with optical coherence tomography (OCT) and fluorescence lifetime imaging (FLIM) (Conference Presentation)

NASA Astrophysics Data System (ADS)

Chen, Xi; Kim, Wihan; Serafino, Michael; Walton, Brian; Jo, Javier A.; Applegate, Brian E.

2017-02-01

We have shown in an ex vivo human coronary artery study that the biochemical information derived from FLIM interpreted in the context of the morphological information from OCT enables a detailed classification of human coronary plaques associated with atherosclerosis. The identification of lipid-rich plaques prone to erosion or rupture and associated with sudden coronary events can impact current clinical practice as well as future development of targeted therapies for "vulnerable" plaques. In order to realize clinical translation of intravascular OCT/FLIM we have had to develop several key technologies. A multimodal catheter endoscope capable of delivering near UV excitation for FLIM and shortwave IR for OCT has been fabricated using a ball lens design with a double clad fiber. The OCT illumination and the FLIM excitation propogate down the inner core while the large outer multimode core captures the fluorescence emission. To enable intravascular pullback imaging with this endoscope we have developed an ultra-wideband fiber optic rotary joint using the same double clad fiber. The rotary joint is based on a lensless design where two cleaved fibers, one fixed and one rotating, are brought into close proximity but not touching. Using water as the lubricant enabled operation over the near UV-shortwave IR range. Transmission over this bandwidth has been measured to be near 100% at rotational frequencies up to 147 Hz. The entire system has been assembled and placed on a mobile cart suitable for cath lab based imaging. System development, performance, and early ex vivo imaging results will be discussed.

Photocleavable Hydrogel-Coated Upconverting Nanoparticles: A Multifunctional Theranostic Platform for NIR Imaging and On-Demand Macromolecular Delivery.

PubMed

Jalani, Ghulam; Naccache, Rafik; Rosenzweig, Derek H; Haglund, Lisbet; Vetrone, Fiorenzo; Cerruti, Marta

2016-01-27

Lanthanide-doped upconverting nanoparticles (UCNPs) have emerged as excellent nanotransducers for converting longer wavelength near-infrared (NIR) light to shorter wavelengths spanning the ultraviolet (UV) to the visible (Vis) regions of the spectrum via a multiphoton absorption process, known as upconversion. Here, we report the development of NIR to UV-Vis-NIR UCNPs consisting of LiYF4:Yb(3+)/Tm(3+)@SiO2 individually coated with a 10 ± 2 nm layer of chitosan (CH) hydrogel cross-linked with a photocleavable cross-linker (PhL). We encapsulated fluorescent-bovine serum albumin (FITC-BSA) inside the gel. Under 980 nm excitation, the upconverted UV emission cleaves the PhL cross-links and instantaneously liberates the FITC-BSA under 2 cm thick tissue. The release is immediately arrested if the excitation source is switched off. The upconverted NIR light allows for the tracking of particles under the tissue. Nucleus pulposus (NP) cells cultured with UCNPs are viable both in the presence and in the absence of laser irradiation. Controlled drug delivery of large biomolecules and deep tissue imaging make this system an excellent theranostic platform for tissue engineering, biomapping, and cellular imaging applications.

Slide-free histology via MUSE: UV surface excitation microscopy for imaging unsectioned tissue (Conference Presentation)

NASA Astrophysics Data System (ADS)

Levenson, Richard M.; Harmany, Zachary; Demos, Stavros G.; Fereidouni, Farzad

2016-03-01

Widely used methods for preparing and viewing tissue specimens at microscopic resolution have not changed for over a century. They provide high-quality images but can involve time-frames of hours or even weeks, depending on logistics. There is increasing interest in slide-free methods for rapid tissue analysis that can both decrease turn-around times and reduce costs. One new approach is MUSE (microscopy with UV surface excitation), which exploits the shallow penetration of UV light to excite fluorescent signals from only the most superficial tissue elements. The method is non-destructive, and eliminates requirement for conventional histology processing, formalin fixation, paraffin embedding, or thin sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography optics. MUSE generates diagnostic-quality histological images that can be rendered to resemble conventional hematoxylin- and eosin-stained samples, with enhanced topographical information, from fresh or fixed, but unsectioned tissue, rapidly, with high resolution, simply and inexpensively. We anticipate that there could be widespread adoption in research facilities, hospital-based and stand-alone clinical settings, in local or regional pathology labs, as well as in low-resource environments.

A new high selective and sensitive turn-on fluorescent and ratiometric absorption chemosensor for Cu2+ based on benzimidazole in aqueous solution and its application in live cell.

PubMed

Bing, Qijing; Wang, Lin; Li, Donglin; Wang, Guang

2018-09-05

A new benzimidazole base turn-on fluorescent and ratiometric absorption chemosensor (L) bearing bidentate ligand for detection of Cu 2+ was designed and synthesized. Fluorescence and UV-vis spectra studies demonstrated that L can detect Cu 2+ ions in aqueous solution using fluorescence enhancement and ratiometric absorption sensing over a wide pH range. Both fluorescent and ratiometric absorption sensing of L for Cu 2+ possessed high selectivity and sensitivity over other competitive metal ions and had low detection limit. Job's plot, mass spectra and DFT calculation indicated the sensing mechanism is the complex formation between L and Cu 2+ with 1:2 stoichiometry. Fluorescence images of HepG2 in the absence and presence of Cu 2+ displayed L had cell permeability and detection ability for Cu 2+ in live cells. Copyright © 2018 Elsevier B.V. All rights reserved.

Methylglyoxal induced glycation and aggregation of human serum albumin: Biochemical and biophysical approach.

PubMed

Ahmed, Azaj; Shamsi, Anas; Khan, Mohd Shahnawaz; Husain, Fohad Mabood; Bano, Bilqees

2018-07-01

Serum protein glycation and formation of advanced glycation end products (AGEs) correlates with many diseases viz. diabetes signifying the importance of studying the glycation pattern of serum proteins. In our present study, methylglyoxal was investigated for its effect on the structure of human serum albumin (HSA); exploring the formation of AGEs and aggregates of HSA. The analytical tools employed includes intrinsic and extrinsic fluorescence, UV spectroscopy, far UV circular dichroism, Thioflavin T fluorescence, congo red binding, polyacrylamide gel electrophoresis (PAGE). UV and fluorescence spectroscopy revealed the structural transition of native HSA evident by new peaks and increased absorbance in UV spectra and quenched fluorescence in the presence of MG. Far UV CD spectroscopy revealed MG induced secondary structural alteration evident by reduced α-helical content. AGEs formation was confirmed by AGEs specific fluorescence. Increased ThT fluorescence and CR absorbance of 10mM MG incubated HSA suggests that glycated HSA results in the formation of aggregates of HSA. SEM and TEM were reported to have an insight of these aggregates. Molecular docking was also utilized to see site specific interaction of MG-HSA. This study is clinically significant as HSA is a clinically relevant protein which plays a crucial role in many diseases. Copyright © 2018 Elsevier B.V. All rights reserved.

Green synthesis of carbon dots originated from Lycii Fructus for effective fluorescent sensing of ferric ion and multicolor cell imaging.

PubMed

Sun, Xiaohan; He, Jiang; Yang, Shenghong; Zheng, Mingda; Wang, Yingying; Ma, Shuang; Zheng, Haipeng

2017-10-01

Green, economical and effective method was developed for synthesis of fluorescent carbon dots (CDs), using one-pot hydrothermal treatment of Lycii Fructus. Optical and structural properties of the CDs have been extensively studied by UV-visible and fluorescence spectroscopic, x-ray diffraction (XRD) techniques, transmission electron microscopy (TEM) and high resolution TEM (HRTEM). Surface functionality and composition of CDs has been illustrated by Fourier transform infrared spectroscopy (FTIR), x-ray photoelectron spectroscopy (XPS) spectra and elemental analysis. The fabricated CDs possess stable fluorescent properties. The fluorescent quantum yield of the CDs can reach 17.2%. The prepared CDs emitted a broad fluorescence between 415 and 545nm and their fluorescence was tuned by changing excitation wavelength. Meanwhile, the fluorescence intensity of the CDs could be significantly quenched by Fe 3+ (turn-off). The CDs exhibit captivating sensitivity and selectivity toward Fe 3+ with a linear range from 0 to 30μM and a detection limit of 21nM. The prepared CDs were successfully applied to the determination of Fe 3+ in the urine samples, the water samples from the from the Yellow River and living HeLa (Henrietta Lacks) cells. Moreover, the low-toxicity and excellent biocompatibility of the CDs were evaluated through MTT assay on HeLa cells. The CDs were also employed as fluorescent probes for multicolor imaging of HeLa cells successfully. Copyright © 2017 Elsevier B.V. All rights reserved.

Toward Fourier interferometry fluorescence excitation/emission imaging of malignant cells combined with photoacoustic microscopy

NASA Astrophysics Data System (ADS)

Kohen, Elli; Hirschberg, Joseph G.; Berry, John P.; Ozkutuk, Nuri; Ornek, Ceren; Monti, Marco; Leblanc, Roger M.; Schachtschabel, Dietrich O.; Haroon, Sumaira

2003-10-01

Dual excitation fluorescence imaging has been used as a first step towards multi-wavelength excitation/emission fluorescence spectral imaging. Target cells are transformed keratinocytes, and other osteosarcoma, human breast and color cancer cells. Mitochondrial membrane potential probes, e.g. TMRM (tetramethylrhodamine methyl ester), Mitotracker Green (Molecular Probes, Inc., Eugene OR,USA; a recently synthesized mitochondrial oxygen probe, [PRE,P1"- pyrene butyl)-2-rhodamine ester] allow dual excitation in the UV plus in teh blue-green spectral regions. Also, using the natural endogenous probe NAD(P)H, preliminary results indicate mitochondrial responses to metabolic challenges (e.g. glucose addition), plus changes in mitochonrial distribution and morphology. In terms of application to biomedicine (for diagnostiscs, prognostsics and drug trials) three parameters have been selected in addition to the natural probe NAD(P)H, i.e. vital fluorescence probing of mitochondria, lysosomes and Golgi apparatus. It is hoped that such a multiparameter approach will allow malignant cell characterization and grading. A new area being introduced is the use of similar methodology for biotechnical applications such as the study of the hydrogen-producing alga Chlamydomonas Reinhardtii, and possible agricultural applications, such as Saccharomyces yeast for oenology. Complementation by Photoacoustic Microscopy is also contemplated, to study the internal conversion component which follows the excitation by photons.

Highly selective rhodamine-based fluorescence turn-on chemosensor for Al3+ ion

NASA Astrophysics Data System (ADS)

Manjunath, Rangasamy; Kannan, Palaninathan

2018-05-01

A new rhodamine-based colorimetric and fluorescent turn-on chemosensor (L) has been designed and synthesized for selective and sensitive detection of Al3+ ion. The sensing behavior toward metal ion was investigated by UV/Vis and fluorescence spectroscopy. Upon addition of Al3+ ion to solution of L provided a visual color change as well as significantly fluorescent enhancement, while other metal ions including Na+, Mg2+, K+, Mn2+, Fe3+, Ni2+, Cu2+, Zn2+, Pb2+, Cd2+ and Hg2+ ions fails to generate a distinct color and spectral changes, the distinct color change and rapid switch-on fluorescence also provide naked eye detection for Al3+ ion. The mechanism involved equilibrium between non-fluorescent spirocyclic form and highly fluorescent ring open form process was utilized and 1:2 stoichiometry for L-Al3+ complex formed with an association constant of 1.42 × 103 M-1. Moreover, chemosensor L was applied for living cell imaging and confirmed that can be used as a fluorescent probe for monitoring Al3+ ion in living cells.

The effect of UV-B radiation on photosynthesis and respiration of phytoplankton, benthic macroalgae and seagrasses.

PubMed

Larkum, A W; Wood, W F

1993-04-01

Several species of marine benthic algae, four species of phytoplankton and two species of seagrass have been subjected to ultraviolet B irradiation for varying lengths of time and the effects on respiration, photosynthesis and fluorescence rise kinetics studied. No effect on respiration was found. Photosynthesis was inhibited to a variable degree in all groups of plants after irradiation over periods of up to 1 h and variable fluorescence was also inhibited in a similar way. The most sensitive plants were phytoplankton and deep-water benthic algae. Intertidal benthic algae were the least sensitive to UV-B irradiation and this may be related to adaptation, through the accumulation of UV-B screening compounds, to high light/high UV-B levels. Inhibition of variable fluorescence (Fv) of the fluorescence rise curve was a fast and sensitive indicator of UV-B damage. Two plants studied, a brown alga and a seagrass, showed very poor recovery of Fv over a period of 32 h.

Quantification of Coffea arabica and Coffea canephora var. robusta concentration in blends by means of synchronous fluorescence and UV-Vis spectroscopies.

PubMed

Dankowska, A; Domagała, A; Kowalewski, W

2017-09-01

The potential of fluorescence, UV-Vis spectroscopies as well as the low- and mid-level data fusion of both spectroscopies for the quantification of concentrations of roasted Coffea arabica and Coffea canephora var. robusta in coffee blends was investigated. Principal component analysis was used to reduce data multidimensionality. To calculate the level of undeclared addition, multiple linear regression (PCA-MLR) models were used with lowest root mean square error of calibration (RMSEC) of 3.6% and root mean square error of cross-validation (RMSECV) of 7.9%. LDA analysis was applied to fluorescence intensities and UV spectra of Coffea arabica, canephora samples, and their mixtures in order to examine classification ability. The best performance of PCA-LDA analysis was observed for data fusion of UV and fluorescence intensity measurements at wavelength interval of 60nm. LDA showed that data fusion can achieve over 96% of correct classifications (sensitivity) in the test set and 100% of correct classifications in the training set, with low-level data fusion. The corresponding results for individual spectroscopies ranged from 90% (UV-Vis spectroscopy) to 77% (synchronous fluorescence) in the test set, and from 93% to 97% in the training set. The results demonstrate that fluorescence, UV, and visible spectroscopies complement each other, giving a complementary effect for the quantification of roasted Coffea arabica and Coffea canephora var. robusta concentration in blends. Copyright © 2017 Elsevier B.V. All rights reserved.

UV superconducting nanowire single-photon detectors with high efficiency, low noise, and 4 K operating temperature

NASA Astrophysics Data System (ADS)

Wollman, E. E.; Verma, V. B.; Beyer, A. D.; Briggs, R. M.; Korzh, B.; Allmaras, J. P.; Marsili, F.; Lita, A. E.; Mirin, R. P.; Nam, S. W.; Shaw, M. D.

2017-10-01

For photon-counting applications at ultraviolet wavelengths, there are currently no detectors that combine high efficiency (> 50%), sub-nanosecond timing resolution, and sub-Hz dark count rates. Superconducting nanowire single-photon detectors (SNSPDs) have seen success over the past decade for photon-counting applications in the near-infrared, but little work has been done to optimize SNSPDs for wavelengths below 400 nm. Here, we describe the design, fabrication, and characterization of UV SNSPDs operating at wavelengths between 250 and 370 nm. The detectors have active areas up to 56 ${\\mu}$m in diameter, 70 - 80% efficiency, timing resolution down to 60 ps FWHM, blindness to visible and infrared photons, and dark count rates of ~ 0.25 counts/hr for a 56 ${\\mu}$m diameter pixel. By using the amorphous superconductor MoSi, these UV SNSPDs are also able to operate at temperatures up to 4.2 K. These performance metrics make UV SNSPDs ideal for applications in trapped-ion quantum information processing, lidar studies of the upper atmosphere, UV fluorescent-lifetime imaging microscopy, and photon-starved UV astronomy.

[Study of the Detecting System of CH4 and SO2 Based on Spectral Absorption Method and UV Fluorescence Method].

PubMed

Wang, Shu-tao; Wang, Zhi-fang; Liu, Ming-hua; Wei, Meng; Chen, Dong-ying; Wang, Xing-long

2016-01-01

According to the spectral absorption characteristics of polluting gases and fluorescence characteristics, a time-division multiplexing detection system is designed. Through this system we can detect Methane (CH4) and sulfur dioxide (SO2) by using spectral absorption method and the SO2 can be detected by using UV fluorescence method. The system consists of four parts: a combination of a light source which could be switched, the common optical path, the air chamber and the signal processing section. The spectral absorption characteristics and fluorescence characteristics are measured first. Then the experiment of detecting CH4 and SO2 through spectral absorption method and the experiment of detecting SO2 through UV fluorescence method are conducted, respectively. Through measuring characteristics of spectral absorption and fluorescence, we get excitation wavelengths of SO2 and CH4 measured by spectral absorption method at the absorption peak are 280 nm and 1.64 μm, respectively, and the optimal excitation wavelength of SO2 measured by UV fluorescence method is 220 nm. we acquire the linear relation between the concentration of CH4 and relative intensity and the linear relation between the concentration of SO2 and output voltage after conducting the experiment of spectral absorption method, and the linearity are 98.7%, 99.2% respectively. Through the experiment of UV fluorescence method we acquire that the relation between the concentration of SO2 and the voltage is linear, and the linearity is 99.5%. Research shows that the system is able to be applied to detect the polluted gas by absorption spectrum method and UV fluorescence method. Combing these two measurement methods decreases the costing and the volume, and this system can also be used to measure the other gases. Such system has a certain value of application.

UV-sensitive scientific CCD image sensors

NASA Astrophysics Data System (ADS)

Vishnevsky, Grigory I.; Kossov, Vladimir G.; Iblyaminova, A. F.; Lazovsky, Leonid Y.; Vydrevitch, Michail G.

1997-06-01

An investigation of probe laser irradiation interaction with substances containing in an environment has long since become a recognized technique for contamination detection and identification. For this purpose, a near and midrange-IR laser irradiation is traditionally used. However, as many works presented on last ecology monitoring conferences show, in addition to traditional systems, rapidly growing are systems with laser irradiation from near-UV range (250 - 500 nm). Use of CCD imagers is one of the prerequisites for this allowing the development of a multi-channel computer-based spectral research system. To identify and analyze contaminating impurities on an environment, such methods as laser fluorescence analysis, UV absorption and differential spectroscopy, Raman scattering are commonly used. These methods are used to identify a large number of impurities (petrol, toluene, Xylene isomers, SO2, acetone, methanol), to detect and identify food pathogens in real time, to measure a concentration of NH3, SO2 and NO in combustion outbursts, to detect oil products in a water, to analyze contaminations in ground waters, to define ozone distribution in the atmosphere profile, to monitor various chemical processes including radioactive materials manufacturing, heterogeneous catalytic reactions, polymers production etc. Multi-element image sensor with enhanced UV sensitivity, low optical non-uniformity, low intrinsic noise and high dynamic range is a key element of all above systems. Thus, so called Virtual Phase (VP) CCDs possessing all these features, seems promising for ecology monitoring spectral measuring systems. Presently, a family of VP CCDs with different architecture and number of pixels is developed and being manufactured. All CCDs from this family are supported with a precise slow-scan digital image acquisition system that can be used in various image processing systems in astronomy, biology, medicine, ecology etc. An image is displayed directly on a PC monitor through a software support.

Correlation fluorescence method of amine detection

NASA Astrophysics Data System (ADS)

Myslitsky, Valentin F.; Tkachuk, Svetlana S.; Rudeichuk, Volodimir M.; Strinadko, Miroslav T.; Slyotov, Mikhail M.; Strinadko, Marina M.

1997-12-01

The amines fluorescence spectra stimulated by UV laser radiation are investigated in this paper. The fluorescence is stimulated by the coherent laser beam with the wavelength 0.337 micrometers . At the sufficient energy of laser stimulation the narrow peaks of the fluorescence spectra are detected besides the wide maximum. The relationship between the fluorescence intensity and the concentration of amines solutions are investigated. The fluorescence intensity temporal dependence on wavelength 0.363 micrometers of the norepinephrine solution preliminarily radiated by UV laser with wavelength 0.337 micrometers was found. The computer stimulated and experimental investigations of adrenaline and norepinephrine mixtures fluorescence spectra were done. The correlation fluorescent method of amines detection is proposed.

All Fiber-Coupled OH Planar Laser-Induced-Fluorescence (OH-PLIF)-Based Two-Dimensional Thermometry.

PubMed

Hsu, Paul S; Jiang, Naibo; Patnaik, Anil K; Katta, Vish; Roy, Sukesh; Gord, James R

2018-04-01

Two-color, planar laser-induced fluorescence (PLIF)-based two-dimensional (2D) thermometry techniques for reacting flows, which are typically developed in the laboratory conditions, face a stiff challenge in their practical implementation in harsh environments such as combustion rigs. In addition to limited optical access, the critical experimental conditions (i.e., uncontrolled humidity, vibration, and large thermal gradients) often restrict sensitive laser system operation and cause difficulties maintaining beam-overlap. Thus, an all fiber-coupled, two-color OH-PLIF system has been developed, employing two long optical fibers allowing isolation of the laser and signal-collection systems. Two OH-excitation laser beams (∼283 nm and ∼286 nm) are delivered through a common 6 m long, 400 µm core, deep ultraviolet (UV)-enhanced multimode fiber. The fluorescence signal (∼310 nm) is collected by a 3 m long, UV-grade imaging fiber. Proof-of-principle temperature measurements are demonstrated in atmospheric pressure, near adiabatic, CH 4 /O 2 /N 2 jet flames. The effects of the excitation pulse interval on fiber transmission are investigated. The proof-of-principle measurements show significant promise for thermometry in harsh environments such as gas turbine engine tests.

NASA Fluid Lensing & MiDAR: Next-Generation Remote Sensing Technologies for Aquatic Remote Sensing

NASA Technical Reports Server (NTRS)

Chirayath, Ved

2018-01-01

We present two recent instrument technology developments at NASA, Fluid Lensing and MiDAR, and their application to remote sensing of Earth's aquatic systems. Fluid Lensing is the first remote sensing technology capable of imaging through ocean waves in 3D at sub-cm resolutions. MiDAR is a next-generation active hyperspectral remote sensing and optical communications instrument capable of active fluid lensing. Fluid Lensing has been used to provide 3D multispectral imagery of shallow marine systems from unmanned aerial vehicles (UAVs, or drones), including coral reefs in American Samoa and stromatolite reefs in Hamelin Pool, Western Australia. MiDAR is being deployed on aircraft and underwater remotely operated vehicles (ROVs) to enable a new method for remote sensing of living and nonliving structures in extreme environments. MiDAR images targets with high-intensity narrowband structured optical radiation to measure an objectâ€"TM"s non-linear spectral reflectance, image through fluid interfaces such as ocean waves with active fluid lensing, and simultaneously transmit high-bandwidth data. As an active instrument, MiDAR is capable of remotely sensing reflectance at the centimeter (cm) spatial scale with a signal-to-noise ratio (SNR) multiple orders of magnitude higher than passive airborne and spaceborne remote sensing systems with significantly reduced integration time. This allows for rapid video-frame-rate hyperspectral sensing into the far ultraviolet and VNIR wavelengths. Previously, MiDAR was developed into a TRL 2 laboratory instrument capable of imaging in thirty-two narrowband channels across the VNIR spectrum (400-950nm). Recently, MiDAR UV was raised to TRL4 and expanded to include five ultraviolet bands from 280-400nm, permitting UV remote sensing capabilities in UV A, B, and C bands and enabling mineral identification and stimulated fluorescence measurements of organic proteins and compounds, such as green fluorescent proteins in terrestrial and aquatic organics.

Evaluation of natural organic matter changes from Lake Hohloh by three-dimensional excitation-emission matrix fluorescence spectroscopy during TiO(2)/UV process.

PubMed

Valencia, Sergio; Marín, Juan M; Restrepo, Gloria; Frimmel, Fritz H

2014-03-15

This study shows the changes of natural organic matter (NOM) from Lake Hohloh, (Black Forest, Germany) during heterogeneous photocatalysis with TiO2 (TiO2/UV). The effect of pH on the adsorption of NOM onto TiO2 in the dark and TiO2/UV degradation of NOM was followed using three-dimensional excitation-emission matrix (EEM) fluorescence. At pH values between 4 and 9, the NOM was adsorbed onto TiO2 in the dark with a greater decrease in the fluorescence intensity and in the spectral shapes, especially under acidic pH conditions. However, at pH = 10 there was not adsorption on NOM which led to a negligible changes the fluorescence intensity. A significant high linear correlation was observed between the DOC adsorption onto TiO2 and the maximum fluorescence intensity. Additionally, the NOM adsorption onto TiO2 and its TiO2/UV degradation shifted the fluorescence maxima toward shorter wavelengths in the EEM contour plots, with a decrease in aromaticity. These changes were accompanied by a substantial decrease in the organically bound halogens adsorbable on activated carbon (AOXFP) and the trihalomethane formation potential (THMFP). Thus, the decrease in maximum fluorescence intensity can be used as an indicator of AOXFP and TTHMFP removal efficiency. Therefore, fluorescence spectroscopy is a robust analytical technique for evaluate TiO2/UV removal of NOM. Copyright © 2013 Elsevier Ltd. All rights reserved.

Homogenization optics to improve detectability of a fluorescence response to a single laser pulse: Detection of feces on apples

USDA-ARS?s Scientific Manuscript database

Fecal contamination of produce is a known food safety risk. Measuring fluorescence responses to UV excitation is an established method for detecting such contamination. One measurement system utilizes a pulsed UV laser to induce a fluorescence response from fecal material and a gated intensified cam...

Biocommunication between Plants and Pollinating Insects through Fluorescence of Pollen and Anthers.

PubMed

Mori, Shinnosuke; Fukui, Hiroshi; Oishi, Masanori; Sakuma, Masayuki; Kawakami, Mari; Tsukioka, Junko; Goto, Katsumi; Hirai, Nobuhiro

2018-06-01

Flowering plants attract pollinators via various stimuli such as odor, color, and shape. Factors determining the foraging behavior of pollinators remain a major theme in ecological and evolutionary research, although the floral traits and cognitive ability of pollinators have been investigated for centuries. Here we show that the autofluorescence emitted from pollen and anthers under UV irradiation may act as another attractant for flower-visiting insects. We have identified fluorescent compounds from pollen and anthers of five plant species as hydroxycinnamoyl derivatives. The fluorescent compounds are also shown to quench UV energy and exhibit antioxidant activity, indicating a function as protectants of pollen genes from UV-induced damage. A two-choice assay using honeybees in the field demonstrated that they perceived the blue fluorescence emitted from the fluorescent compounds and were attracted to it. This result suggested that the fluorescence from pollen and anthers serves as a visual cue to attract pollinators under sunlight.

Double-Protected All-Inorganic Perovskite Nanocrystals by Crystalline Matrix and Silica for Triple-Modal Anti-Counterfeiting Codes.

PubMed

Xu, Leimeng; Chen, Jiawei; Song, Jizhong; Li, Jianhai; Xue, Jie; Dong, Yuhui; Cai, Bo; Shan, Qingsong; Han, Boning; Zeng, Haibo

2017-08-09

Novel fluorescence with highly covert and reliable features is quite desirable to combat the sophisticated counterfeiters. Herein, we report a simultaneously triple-modal fluorescent characteristic of CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 by the excitation of thermal, ultraviolet (UV) and infrared (IR) light for the first time, which can be applied for the multiple modal anti-counterfeiting codes. The diphasic structure CsPbBr 3 @Cs 4 PbBr 6 nanocrystals (NCs) was synthesized via the typical reprecipitation method followed by uniformly encapsulation into silica microspheres. Cubic CsPbBr 3 is responsible for the functions of anti-counterfeiting, while Cs 4 PbBr 6 crystalline and SiO 2 are mainly to protect unstable CsPbBr 3 NCs from being destroyed by ambient conditions. The as-prepared CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 NCs possess improved stability and are capable of forming printable ink with organic binders for patterns. Interestingly, the fluorescence of diphasic CsPbBr 3 @Cs 4 PbBr 6 /SiO 2 capsule patterns can be reversibly switched by the heating, UV, and IR light irradiation, which has been applied as triple-modal fluorescent anti-counterfeiting codes. The results demonstrate that the perovskite@silica capsules are highly promising for myriad applications in areas such as fluorescent anti-counterfeiting, optoelectronic devices, medical diagnosis, and biological imaging.

Development of a Technique for Separating Raman Scattering Signals from Background Emission with Single-Shot Measurement Potential

NASA Technical Reports Server (NTRS)

Hartfield, Roy J., Jr.; Dobson, Chris; Eskridge, Richard; Wehrmeyer, Joseph A.

1997-01-01

A novel technique for extracting Q-branch Raman signals scattered by a diatomic species from the emission spectrum resulting from the irradiation of combustion products using a broadband excimer laser has been developed. This technique is based on the polarization characteristics of vibrational Raman scattering and can be used for both single-shot Raman extraction and time-averaged data collection. The Q-branch Raman signal has a unique set of polarization characteristics which depend on the direction of the scattering while fluorescence signals are unpolarized. For the present work, a calcite crystal is used to separate the horizonal component of a collected signal from the vertical component. The two components are then sent through a UV spectrometer and imaged onto an intensified CCD camera separately. The vertical component contains both the Raman signal and the interfering fluorescence signal. The horizontal component contains the fluorescence signal and a very weak component of the Raman signal; hence, the Raman scatter can be extracted by taking the difference between the two signals. The separation of the Raman scatter from interfering fluorescence signals is critically important to the interpretation of the Raman for cases in which a broadband ultraviolet (UV) laser is used as an excitation source in a hydrogen-oxygen flame and in all hydrocarbon flames. The present work provides a demonstration of the separation of the Raman scatter from the fluorescence background in real time.

Facile patterning of hybrid CdSe nanoparticle films by photoinduced surface defects.

PubMed

Park, Yushin; Felipe, Mary Jane; Advincula, Rigoberto C

2011-11-01

The photopatterning of CdSe quantum dots (QDs) films is facilitated by preparing defect-rich QDs on selective sites on the film. A key step is UV irradiation in the presence of a polar solvent such as methanol in situ as a "developer" which readily dissolves trioctylphosphine oxide (TOPO) but not the QDs. This results in a dramatically reduced photopatterning time and irradiation intensity requirement. The optical property changes were examined by UV-vis and fluorescence spectroscopy. Furthermore, the photo-oxidized pattern of the CdSe QD film was readily observed by fluorescence microscopy. The chemical change due to attenuation of the P═O vibration of TOPO (due to its removal) could be detected by FT-IR imaging or FT-IR chemical mapping. Thus, the protocol is a simple yet effective way of patterning PL properties of QD films at much reduced exposure time compared to previously reported methods. It may find utility for a host of cell-based film assays and PL display device applications at various resolutions.

Synthesis and characterization of the fluorescent probes for the labeling of Microthrix parvicella.

PubMed

Li, Songya; Fei, Xuening; Jiao, Xiumei; Lin, Dayong; Zhang, Baolian; Cao, Lingyun

2016-03-01

Although the fluorescent in situ hybridization (FISH) has been widely used to identify the Microthrix parvicella (M. parvicella), there are a few disadvantages and difficulties, such as complicated process, time consuming, etc. In this work, a series of fluorescent probes, which were modified by long-chain alkane with hydrophobic property and based on the property of M. parvicella utilizing long-chain fatty acids (LCFA), for the labeling of M. parvicella in bulking sludge were designed, synthesized, and characterized. The probes were characterized by ultraviolet-visible (UV-Vis) absorption spectra, fluorescence spectra, (1)H NMR spectra, and mass spectra, and the photostability and hydrophobic property of probes were investigated. All the results showed that the probes were quite stable and suitable for the fluorescent labeling. The probes had a large stoke shift of 98-137 nm, which was benefit for the fluorescent labeling. In the fluorescent labeling of M. parvicella by the synthesized probes, the probes had excellent labeling effects. By comparison of the images and the Image Pro Plus 6.0 analysis, the optimal concentration of the probes in the activated sludge sample for labeling was 0.010 mmol/L and the probe 3d had the best labeling. In addition, the effect of the duration time of probes was also investigated, and the results showed that the fluorescent intensity of probes hardly changed in a long period of time and it was suitable for labeling.

Chemometric classification of Chinese lager beers according to manufacturer based on data fusion of fluorescence, UV and visible spectroscopies.

PubMed

Tan, Jin; Li, Rong; Jiang, Zi-Tao

2015-10-01

We report an application of data fusion for chemometric classification of 135 canned samples of Chinese lager beers by manufacturer based on the combination of fluorescence, UV and visible spectroscopies. Right-angle synchronous fluorescence spectra (SFS) at three wavelength difference Δλ=30, 60 and 80 nm and visible spectra in the range 380-700 nm of undiluted beers were recorded. UV spectra in the range 240-400 nm of diluted beers were measured. A classification model was built using principal component analysis (PCA) and linear discriminant analysis (LDA). LDA with cross-validation showed that the data fusion could achieve 78.5-86.7% correct classification (sensitivity), while those rates using individual spectroscopies ranged from 42.2% to 70.4%. The results demonstrated that the fluorescence, UV and visible spectroscopies complemented each other, yielding higher synergic effect. Copyright © 2015 Elsevier Ltd. All rights reserved.

Bioconvertible vitamin antioxidants improve sunscreen photoprotection against UV-induced reactive oxygen species.

PubMed

Hanson, Kerry M; Clegg, Robert M

2003-01-01

The ability of sunscreens and antioxidants to deactivate highly destructive reactive oxygen species in human skin has remained inconclusive. Two-photon fluorescence imaging microscopy was used to determine the effect of sunscreen/antioxidant combinations upon UV-induced ROS generation in ex vivo human skin. A sunscreen combination containing octylmethoxycinnamate (Parsol MCX) and avobenzone (Parsol 1789) at SPF 8 and SPF 15 was tested for its ability to prevent UV radiation from generating ROS in the viable epidermal strata of ex vivo human skin. A UV dose equivalent to two hours of North American solar UV was used to irradiate the skin. Each sunscreen reduced the amount of ROS induced in the viable strata by a value consistent with the SPF level. UV photons that were not absorbed/scattered by the sunscreen formulations generated ROS within the viable epidermal layers. The addition of the bioconvertible antioxidants vitamin E acetate and sodium ascorbyl phosphate (STAY-C 50) improves photoprotection by converting to vitamins E and C, respectively, within the skin. The bioconversion forms an antioxidant reservoir that deactivates the ROS generated (within the strata granulosum, spinosum, and basale) by the UV photons that the sunscreens do not block in the stratum corneum.

Magnetic Field Effects on Triplet-Triplet Annihilation in Solutions: Modulation of Visible/NIR Luminescence.

PubMed

Mani, Tomoyasu; Vinogradov, Sergei A

2013-08-06

Photon upconversion based on sensitized triplet-triplet annihilation (TTA) presents interest for such areas as photovoltaics and imaging. Usually energy upconversion is observed as p -type delayed fluorescence from molecules whose triplet states are populated via energy transfer from a suitable triplet donor, followed by TTA. Magnetic field effects (MFE) on delayed fluorescence in molecular crystals are well known; however, there exist only a few examples of MFE on TTA in solutions, and all of them are limited to UV-emitting materials. Here we present MFE on TTA-mediated visible and near infrared (NIR) emission, sensitized by far-red absorbing metalloporphyrins in solutions at room temperature. In addition to visible delayed fluorescence from annihilator, we also observed NIR emission from the sensitizer, occurring as a result of triplet-triplet energy transfer back from annihilator, termed "delayed phosphorescence". This emission also exhibits MFE, but opposite in sign to the annihilator fluorescence.

An imaging system for PLIF/Mie measurements for a combusting flow

NASA Technical Reports Server (NTRS)

Wey, C. C.; Ghorashi, B.; Marek, C. J.; Wey, C.

1990-01-01

The equipment required to establish an imaging system can be divided into four parts: (1) the light source and beam shaping optics; (2) camera and recording; (3) image acquisition and processing; and (4) computer and output systems. A pulsed, Nd:YAG-pummped, frequency-doubled dye laser which can freeze motion in the flowfield is used for an illumination source. A set of lenses is used to form the laser beam into a sheet. The induced fluorescence is collected by an UV-enhanced lens and passes through an UV-enhanced microchannel plate intensifier which is optically coupled to a gated solid state CCD camera. The output of the camera is simultaneously displayed on a monitor and recorded on either a laser videodisc set of a Super VHS VCR. This videodisc set is controlled by a minicomputer via a connection to the RS-232C interface terminals. The imaging system is connected to the host computer by a bus repeater and can be multiplexed between four video input sources. Sample images from a planar shear layer experiment are presented to show the processing capability of the imaging system with the host computer.

UV red fluorescence of Eubacterium lentum.

PubMed Central

Mosca, A; Strong, C A; Finegold, S M

1993-01-01

Twenty-nine clinical isolates of Eubacterium lentum and two type species were evaluated for the ability to fluoresce under UV light. Twenty-one of the 29 isolates and both of the reference strains showed orange-to-red fluorescence. This fluorescence did not require blood or hemin in the culture media and did not fade upon air exposure. The fluorescent pigment, after extraction by 1 N NaOH, showed peak excitation at a wavelength of around 400 nm. The capacity of E. lentum to produce fluorescence may be a useful and time-sparing laboratory aid for its identification. PMID:8463378

Atomic force imaging microscopy investigation of the interaction of ultraviolet radiation with collagen thin films

NASA Astrophysics Data System (ADS)

Stylianou, A.; Yova, D.; Alexandratou, E.; Petri, A.

2013-02-01

Collagen is the major fibrous protein in the extracellular matrix and consists a significant component of skin, bone, cartilage and tendon. Due to its unique properties, it has been widely used as scaffold or culture substrate for tissue regeneration or/and cell-substrate interaction studies. The ultraviolet light-collagen interaction investigations are crucial for the improvement of many applications such as that of the UV irradiation in the field of biomaterials, as sterilizing and photo-cross-linking method. The aim of this paper was to investigate the mechanisms of UV-collagen interactions by developing a collagen-based, well characterized, surface with controlled topography of collagen thin films in the nanoscale range. The methodology was to quantify the collagen surface modification induced on ultraviolet radiation and correlate it with changes induced in cells. Surface nanoscale characterization was performed by Atomic Force Microscopy (AFM) which is a powerful tool and offers quantitative and qualitative information with a non-destructive manner. In order to investigate cells behavior, the irradiated films were used for in vitro cultivation of human skin fibroblasts and the cells morphology, migration and alignment were assessed with fluorescence microscopy imaging and image processing methods. The clarification of the effects of UV light on collagen thin films and the way of cells behavior to the different modifications that UV induced to the collagen-based surfaces will contribute to the better understanding of cell-matrix interactions in the nanoscale and will assist the appropriate use of UV light for developing biomaterials.

Application of Time-Resolved Spectroscopies to the Study of Energetic Materials - 1982

DTIC Science & Technology

1983-05-24

fluores- cence intensity as a function of UV pulse energy, for individual laser shots. The lower curve shows the UV + probe induced fluorescence... intensity as a function of UV pulse energy, for individual laser shots. The lower curve shows the UV + probe Induced fluorescence, at 1 ns delay...locked Nd:YAG Laser Pulse ", Appl. Phys. Lett 26, 501-503 (1975). 97 43. A. J. Campillo, V. H. Kollman and S. L. Shapiro, " Intensity Dependence of

Technical Testing of Deep-UV Solid-State Sources for Fluorescence Lifetime Measurements in the Frequency Domain

DTIC Science & Technology

2007-02-01

fluxes at wavelengths short enough for excitation of fluorescence in basic biological fluorophores and bacterial agents. In particular, deep- UV LEDs ...can be used for excitation of aromatic amino acids, whereas near- UV LEDs are suitable for excitation of autofluorescent coenzymes. The SUVOS AlGaN... LEDs as well as commercial InGaN near- UV LEDs were tested for spectral purity and the possibility of high-frequency modulation up to 200 MHz and

Red fluorescence increases with depth in reef fishes, supporting a visual function, not UV protection

PubMed Central

Meadows, Melissa G.; Anthes, Nils; Dangelmayer, Sandra; Alwany, Magdy A.; Gerlach, Tobias; Schulte, Gregor; Sprenger, Dennis; Theobald, Jennifer; Michiels, Nico K.

2014-01-01

Why do some marine fishes exhibit striking patterns of natural red fluorescence? In this study, we contrast two non-exclusive hypotheses: (i) that UV absorption by fluorescent pigments offers significant photoprotection in shallow water, where UV irradiance is strongest; and (ii) that red fluorescence enhances visual contrast at depths below −10 m, where most light in the ‘red’ 600–700 nm range has been absorbed. Whereas the photoprotection hypothesis predicts fluorescence to be stronger near the surface and weaker in deeper water, the visual contrast hypothesis predicts the opposite. We used fluorometry to measure red fluorescence brightness in vivo in individuals belonging to eight common small reef fish species with conspicuously red fluorescent eyes. Fluorescence was significantly brighter in specimens from the −20 m sites than in those from −5 m sites in six out of eight species. No difference was found in the remaining two. Our results support the visual contrast hypothesis. We discuss the possible roles fluorescence may play in fish visual ecology and highlight the possibility that fluorescent light emission from the eyes in particular may be used to detect cryptic prey. PMID:25030989

Selective and sensitive fluorescent sensor for Pd2+ using coumarin 460 for real-time and biological applications.

PubMed

Ashwin, Bosco Christin Maria Arputham; Sivaraman, Gandhi; Stalin, Thambusamy; Yuvakkumar, Rathinam; Muthu Mareeswaran, Paulpandian

2018-06-01

The efficient fluorescent property of coumarin 460 (C460) is utilized to sense the Pd 2+ selectively and sensitively. Fabrication of a sensor strip using commercial adhesive tape is achieved and the detection of Pd 2+ is attempted using a handy UV torch. The naked eye detection in solution state using UV chamber is also attempted. The calculated high binding constant values support the strong stable complex formation of Pd 2+ with C460. The detection limit up to 2.5 × 10 -7  M is achieved using fluorescence spectrometer, which is considerably low from the WHO's recommendation. The response of coumarin 460 with various cations also studied. The quenching is further studied by the lifetime measurements. The binding mechanism is clearly explained by the 1 H NMR titration. The sensing mechanism is established as ICT. C460 strip's Pd 2+ quenching detection is further confirmed by solid-state PL study. The in-vitro response of Pd 2+ in a living cell is also studied using fluorescent imaging studies by means of HeLa cell lines and this probe is very compatible with biological environments. It could be applicable to sense trace amounts of a Pd 2+ ion from various industries. Compared with previous reports, this one is very cheap, sensitive, selective and suitable for biological systems. Copyright © 2018 Elsevier B.V. All rights reserved.

Efficient synthesis of highly fluorescent nitrogen-doped carbon dots for cell imaging using unripe fruit extract of Prunus mume

NASA Astrophysics Data System (ADS)

Atchudan, Raji; Edison, Thomas Nesakumar Jebakumar Immanuel; Sethuraman, Mathur Gopalakrishnan; Lee, Yong Rok

2016-10-01

Highly fluorescent nitrogen-doped carbon dots (N-CDs) were synthesized using the extract of unripe Prunus mume (P. mume) fruit by a simple one step hydrothermal-carbonization method. The N-CDs were synthesized at different pH ranges, 2.3, 5, 7, and 9. The pH of the P. mume extract was adjusted using an aqueous ammonia solution (25%). The optical properties of N-CDs were examined by UV-vis and fluorescence spectroscopy. The N-CDs synthesized at pH 9 emitted high fluorescence intensity compared to other obtained N-CDs. The N-CDs synthesized at pH 9 was further characterized by high resolution transmission electron microscopy (HR-TEM), X-ray diffraction (XRD), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and Fourier transform-infra red (FT-IR) spectroscopy. HR-TEM showed that the average size of the synthesized N-CDs was approximately 9 nm and the interlayer distance was 0.21 nm, which was validated by XRD. The graphitic nature of the synthesized N-CDs were confirmed by Raman spectroscopy. XPS and FT-IR spectroscopy confirmed the doping of the nitrogen moiety over the synthesized CDs. The synthesized nitrogen doped CDs (N-CDs) were low toxicity and were used as a staining probe for fluorescence cell imaging.

Nonlinear Optical Characterization of Membrane Protein Microcrystals and Nanocrystals.

PubMed

Newman, Justin A; Simpson, Garth J

2016-01-01

Nonlinear optical methods such as second harmonic generation (SHG) and two-photon excited UV fluorescence (TPE-UVF) imaging are promising approaches to address bottlenecks in the membrane protein structure determination pipeline. The general principles of SHG and TPE-UVF are discussed here along with instrument design considerations. Comparisons to conventional methods in high throughput crystallization condition screening and crystal quality assessment prior to X-ray diffraction are also discussed.

UV/blue light-induced fluorescence for assessing apple maturity

NASA Astrophysics Data System (ADS)

Noh, Hyun Kwon; Lu, Renfu

2005-11-01

Chlorophyll fluorescence has been researched for assessing fruit post-harvest quality and condition. The objective of this preliminary research was to investigate the potential of fluorescence spectroscopy for measuring apple fruit quality. Ultraviolet (UV) and blue light was used as an excitation source for inducing fluorescence in apples. Fluorescence spectra were measured from 'Golden Delicious' (GD) and 'Red Delicious' (RD) apples by using a visible/near-infrared spectrometer after one, three, and five minutes of continuous UV/blue light illumination. Standard destructive tests were performed to measure fruit firmness, skin and flesh color, soluble solids and acid content from the apples. Calibration models for each of the three illumination time periods were developed to predict fruit quality indexes. The results showed that fluorescence emission decreased steadily during the first three minutes of UV/blue light illumination and was stable within five minutes. The differences were minimal in the model prediction results based on fluorescence data at one, three or five minutes of illumination. Overall, better predictions were obtained for apple skin chroma and hue and flesh hue with values for the correlation coefficient of validation between 0.80 and 0.90 for both GD and RD. Relatively poor predictions were obtained for fruit firmness, soluble solids content, titrational acid, and flesh chroma. This research demonstrated that fluorescence spectroscopy is potentially useful for assessing selected quality attributes of apple fruit and further research is needed to improve fluorescence measurements so that better predictions of fruit quality can be achieved.

Influence of the ionic liquid 1-butyl-3-methylimidazolium bromide on amyloid fibrillogenesis in lysozyme: Evidence from photophysical and imaging studies.

PubMed

Basu, Anirban; Bhattacharya, Subhash Chandra; Kumar, Gopinatha Suresh

2018-02-01

Many proteins can abnormally fold to form pathological amyloid deposits/aggregates that are responsible for various degenerative disorders called amyloidosis. Here we have examined the anti-amyloidogenic potency of an ionic liquid, 1-butyl-3-methylimidazolium bromide, using lysozyme as a model system. Thioflavin T fluorescence assay demonstrated that the ionic liquid suppressed the formation of lysozyme fibrils significantly. This observation was further confirmed by the Congo red assay. Fluorescence microscopy, intrinsic fluorescence studies, nile red fluorescence assay, ANS binding assay and circular dichroism studies also testified diminishing of the fibrillogenesis in the presence of ionic liquid. Formation of amyloid fibrils was also characterized by α to β conformational transition. From far-UV circular dichroism studies it was observed that the β-sheet content of the lysozyme samples decreased in the presence of the ionic liquid which in turn implied that fibrillogenesis was supressed by the ionic liquid. Atomic force microscopy imaging unequivocally established that the ionic liquid attenuated fibrillogenesis in lysozyme. These results may be useful for the development of more effective therapeutics for amyloidosis. Copyright © 2017 Elsevier B.V. All rights reserved.

A simple and sensitive high-throughput GFP screening in woody and herbaceous plants.

PubMed

Hily, Jean-Michel; Liu, Zongrang

2009-03-01

Green fluorescent protein (GFP) has been used widely as a powerful bioluminescent reporter, but its visualization by existing methods in tissues or whole plants and its utilization for high-throughput screening remains challenging in many species. Here, we report a fluorescence image analyzer-based method for GFP detection and its utility for high-throughput screening of transformed plants. Of three detection methods tested, the Typhoon fluorescence scanner was able to detect GFP fluorescence in all Arabidopsis thaliana tissues and apple leaves, while regular fluorescence microscopy detected it only in Arabidopsis flowers and siliques but barely in the leaves of either Arabidopsis or apple. The hand-held UV illumination method failed in all tissues of both species. Additionally, the Typhoon imager was able to detect GFP fluorescence in both green and non-green tissues of Arabidopsis seedlings as well as in imbibed seeds, qualifying it as a high-throughput screening tool, which was further demonstrated by screening the seedlings of primary transformed T(0) seeds. Of the 30,000 germinating Arabidopsis seedlings screened, at least 69 GFP-positive lines were identified, accounting for an approximately 0.23% transformation efficiency. About 14,000 seedlings grown in 16 Petri plates could be screened within an hour, making the screening process significantly more efficient and robust than any other existing high-throughput screening method for transgenic plants.

Fluorescent minerals - A potential source of UV protection and visible light for the growth of green algae and cyanobacteria in extreme cosmic environments

NASA Astrophysics Data System (ADS)

Omairi, Tareq; Wainwright, Milton

2015-07-01

We propose that green algae (Chlorella variabilis and Dunaliella tertiolecta) and cyanobacteria (Synechococcus elongatus and Nostoc commune) can grow inside fluorescent rock minerals which convert damaging UV light to visible light, thereby allowing these organisms to survive and thrive in UV-rich environments without (or with limited) visible light, which would otherwise be inimical to them. The four microorganisms were incubated inside fluorescent rocks composed of fluorite, calcite and pyrite. The resultant growth was then measured following exposure to UV radiation, with the use of optical density and measurement of chlorophyll concentration. Results show that the microorganisms were shielded from harmful UV in these semi-transparent rocks, while at the same time benefiting from the fact that the minerals converted UV to visible light; this have been shown by a statistically significant increase in their growth, which although lower than when the cells were incubated in sunlight, was significantly higher than in controls incubated in the dark.

Photomultiplier window materials under electron irradiation - Fluorescence and phosphorescence

NASA Technical Reports Server (NTRS)

Viehmann, W.; Eubanks, A. G.; Pieper, G. F.; Bredekamp, J. H.

1975-01-01

The fluorescence and phosphorescence of photomultiplier window materials under electron irradiation have been investigated using a Sr-90/Y-90 beta emitter as the electron source. Spectral emission curves of UV-grade, optical-grade, and electron-irradiated samples of MgF2 and LiF, and of CaF2, BaF2, sapphire, fused silica, and UV-transmitting glasses were obtained over the 200-650-nm spectral range. Fluorescence yields were determined on these materials utilizing photomultiplier tubes with cesium telluride, bialkali, and trialkali (S-20) photocathodes, respectively. Optical-grade MgF2 and LiF, as well as electron-irradiated UV-grade samples of these two materials, show enhanced fluorescence due to color-center formation and associated emission bands in the blue and red wavelength regions. Large variations in fluorescence intensities were found in UV-grade sapphire samples of different origins, particularly in the red end of the spectrum, presumably due to various amounts of chromium-ion content. Phosphorescence decay with time is best described by a sum of exponential terms, with time constants ranging from a few minutes to several days.

Photoluminescent ZnO Nanoparticles and Their Biological Applications

PubMed Central

Zhang, Zheng-Yong; Xiong, Huan-Ming

2015-01-01

During the past decades, numerous achievements concerning luminescent zinc oxide nanoparticles (ZnO NPs) have been reported due to their improved luminescence and good biocompatibility. The photoluminescence of ZnO NPs usually contains two parts, the exciton-related ultraviolet (UV) emission and the defect-related visible emission. With respect to the visible emission, many routes have been developed to synthesize and functionalize ZnO NPs for the applications in detecting metal ions and biomolecules, biological fluorescence imaging, nonlinear multiphoton imaging, and fluorescence lifetime imaging. As the biological applications of ZnO NPs develop rapidly, the toxicity of ZnO NPs has attracted more and more attention because ZnO can produce the reactive oxygen species (ROS) and release Zn2+ ions. Just as a coin has two sides, both the drug delivery and the antibacterial effects of ZnO NPs become attractive at the same time. Hence, in this review, we will focus on the progress in the synthetic methods, luminescent properties, and biological applications of ZnO NPs.

Imaging camera system of OYGBR-phosphor-based white LED lighting

NASA Astrophysics Data System (ADS)

Kobashi, Katsuya; Taguchi, Tsunemasa

2005-03-01

The near-ultraviolet (nUV) white LED approach is analogous to three-color fluorescent lamp technology, which is based on the conversion of nUV radiation to visible light via the photoluminescence process in phosphor materials. The nUV light is not included in the white light generation from nUV-based white LED devices. This technology can thus provide a higher quality of white light than the blue and YAG method. A typical device demonstrates white luminescence with Tc=3,700 K, Ra > 93, K > 40 lm/W and chromaticity (x, y) = (0.39, 0.39), respectively. The orange, yellow, green and blue OYGB) or orange, yellow, red, green and blue (OYRGB) device shows a luminescence spectrum broader than of an RGB white LED and a better color rendering index. Such superior luminous characteristics could be useful for the application of several kinds of endoscope. We have shown the excellent pictures of digestive organs in a stomach of a dog due to the strong green component and high Ra.

Roles of dynamin-related protein 1 in the regulation of mitochondrial fission and apoptosis in response to UV stimuli

NASA Astrophysics Data System (ADS)

Zhang, Zhenzhen; Feng, Jie; Wu, Shengnan

2011-03-01

Mitochondria are dynamic structures that frequently divide and fuse with one another to form interconnecting network. This network disintegrates into punctiform organelles during apoptosis. However, it remains unclear whether this event has a significant impact on the rate of cell death or only accompanies apoptosis as an epiphenomenon. In this study, we investigate the role of dynamin-related protein 1 (Drp1), a large GTPase that mediates outer mitochondrial membrane fission, in mitochondrial morphology and apoptosis in response to UV irradiation in human lung adenocarcinoma cells (ASTC-a-1) and HeLa cells. Using time-lapse fluorescent imaging, we find that Drp1 primarily distributes in cytosol under physiological conditions. After UV treatment, Drp1 translocates from cytosol to mitochondria, indicating the enhancement of Drp1 mitochondrial accumulation. Down-regulation of Drp1 by shRNA inhibits UV-induced apoptosis. Our results suggest that Drp1 is involved in the regulation of transition from a reticulo-tubular to a punctiform mitochondrial phenotype and mitochondrial fission plays an important role in UV-induced apoptosis.

Potential of BODIPY-cholesterol for analysis of cholesterol transport and diffusion in living cells.

PubMed

Wüstner, Daniel; Lund, Frederik W; Röhrl, Clemens; Stangl, Herbert

2016-01-01

Cholesterol is an abundant and important lipid component of cellular membranes. Analysis of cholesterol transport and diffusion in living cells is hampered by the technical challenge of designing suitable cholesterol probes which can be detected for example by optical microscopy. One strategy is to use intrinsically fluorescent sterols, as dehydroergosterol (DHE), having minimal chemical alteration compared to cholesterol but giving low fluorescence signals in the UV region of the spectrum. Alternatively, one can use dye-tagged cholesterol analogs and in particular BODIPY-cholesterol (BChol), whose synthesis and initial characterization was pioneered by Robert Bittman. Here, we give a general overview of the properties and applications but also limitations of BODIPY-tagged cholesterol probes for analyzing intracellular cholesterol trafficking. We describe our own experiences and collaborative efforts with Bob Bittman for studying diffusion in the plasma membrane (PM) and uptake of BChol in a quantitative manner. For that purpose, we used a variety of fluorescence approaches including fluorescence correlation spectroscopy and its imaging variants, fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP). We also describe pulse-chase studies from the PM using BChol in direct comparison to DHE. Based on the gathered imaging data, we present a two-step kinetic model for sterol transport between PM and recycling endosomes. In addition, we highlight the suitability of BChol for determining transport of lipoprotein-derived sterol using electron microscopy (EM) and show that this approach ideally complements fluorescence studies. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

UV-fluorescence spectroscopic technique in the diagnosis of breast, ovarian, uterus, and cervix cancer

NASA Astrophysics Data System (ADS)

Das, Bidyut B.; Glassman, Wenling S.; Alfano, Robert R.; Cleary, Joseph; Prudente, R.; Celmer, Edward J.; Lubicz, Stephanie

1991-06-01

Malignant breast tumors can be separated from benign and normal tissues using uv-fluorescence spectroscopic technique. Using the same method one can also distinguish cancerous tissues from noncancerous ones in case of cervix, uterus and ovary.

Single-photon counting multicolor multiphoton fluorescence microscope.

PubMed

Buehler, Christof; Kim, Ki H; Greuter, Urs; Schlumpf, Nick; So, Peter T C

2005-01-01

We present a multicolor multiphoton fluorescence microscope with single-photon counting sensitivity. The system integrates a standard multiphoton fluorescence microscope, an optical grating spectrograph operating in the UV-Vis wavelength region, and a 16-anode photomultiplier tube (PMT). The major technical innovation is in the development of a multichannel photon counting card (mC-PhCC) for direct signal collection from multi-anode PMTs. The electronic design of the mC-PhCC employs a high-throughput, fully-parallel, single-photon counting scheme along with a high-speed electrical or fiber-optical link interface to the data acquisition computer. There is no electronic crosstalk among the detection channels of the mC-PhCC. The collected signal remains linear up to an incident photon rate of 10(8) counts per second. The high-speed data interface offers ample bandwidth for real-time readout: 2 MByte lambda-stacks composed of 16 spectral channels, 256 x 256 pixel image with 12-bit dynamic range can be transferred at 30 frames per second. The modular design of the mC-PhCC can be readily extended to accommodate PMTs of more anodes. Data acquisition from a 64-anode PMT has been verified. As a demonstration of system performance, spectrally resolved images of fluorescent latex spheres and ex-vivo human skin are reported. The multicolor multiphoton microscope is suitable for highly sensitive, real-time, spectrally-resolved three-dimensional imaging in biomedical applications.

Studies on activated cytostatic fluorouracil as photosensitizer: to use in eye tumor treatment

NASA Astrophysics Data System (ADS)

Pascu, Mihail-Lucian; Carstocea, Benone D.; Brezeanu, Mihail; Voicu, Letitia; Staicu, Angela; Gazdaru, Doina M.; Pascu, Ruxandra A.

2004-09-01

Hydroxypyrimidine 5-fluorouracil (5-FU) belongs to the cytostatics group known as antimetabolites. The effect of UV irradiation on 5-FU was investigated by absorption and fluorescence spectroscopy. The study of the photosensitizer properties of 5-FU was made since their effects could be enhanced by exposure to UV radiation at different doses. Solutions 2.5x10-4M in natural saline water (0.8% NaCl), irradiated with optical beams emitted by N2 laser and UV Hg classic lamp, were used. The 5-FU was chosen due to its strong absorption along a large spectral range which makes possible the fluorescence excitation in UV. The absorption spectra exhibit bands between 250 - 450 nm. The emission fluorescence was measured in the 400-550 nm spectral range, with λex=320 and 350 nm for samples irradiated with Hg lamp and with λex=360 nm for samples irradiated with N2 laser. The excitation fluorescence was measured in the spectral range 200-400 nm, with λem=440 nm for samples irradiated with N2 laser. The spectra reveal a fluorescence enhancement with the exposure time, with a maximum at 3 min due to the transformation of 5-FU molecule into a fluorescent tautomeric form. The destruction more rapid than usual of the neovascularisation was observed for conjunctive of rabbit eyes, when they are impregnated with 5-FU solution and exposed to incoherent UV and visible light.

Upconverting fluorescent nanoparticles for biodetection and photoactivation

NASA Astrophysics Data System (ADS)

Huang, Kai; Li, WenKai; Jayakumar, Muthu Kumara Gnanasammandhan; Zhang, Yong

2013-03-01

Fluorophores including fluorescent dyes/proteins and quantum dots (QDs) are used for fluorescence-based imaging and detection. These are based on `downconversion fluorescence' and have several drawbacks: photobleaching, autofluorescence, short tissue penetration depth and tissue photo-damage. Upconversion fluorescent nanoparticles (UCNs) emit detectable photons of higher energy in the short wavelength range upon irradiation with near-infrared (NIR) light based on a process termed `upconversion'. UCNs show absolute photostability, negligible autofluorescence, high penetration depth and minimum photodamage to biological tissues. Lanthanide doped nanocrystals with nearinfrared NIR-to-NIR and/or NIR-to-VIS and/or NIR-to-UV upconversion fluorescence emission have been synthesized. The nanocrystals with small size and tunable multi-color emission have been developed. The emission can be tuned by doping different upconverting lanthanide ions into the nanocrystals. The nanocrystals with core-shell structure have also been prepared to tune the emission color. The surfaces of these nanocrystals have been modified to render them water dispersible and biocompatible. They can be used for ultrasensitive interference-free biodetection because most biomolecules do not have upconversion properties. UCNs are also useful for light based therapy with enhanced efficiency, for example, photoactivation.

Nontoxic fluorescent carbon nanodot serving as a light conversion material in plant for UV light utilization.

PubMed

Sai, Liman; Liu, Siqi; Qian, Xuexue; Yu, Yahui; Xu, Xiaofeng

2018-05-21

In this study, water-soluble fluorescent carbon nanodots (CNDs) were directly injected into the leaf of nicotiana tabacum. With the help of UV-to-blue light conversion nanomaterial, the photosynthetic rate of the leaf was improved 18% upon additional 6 W UV irradiation. The photostability and toxicity of different kinds of CNDs were discussed. The results showed that CNDs functionalized with NH 2 -groups on their surfaces could maintain good fluorescence in plant leaf, and CNDs with complex surface groups tended to have high toxicity to the plant. The NH 2 -functionalized CNDs with non-toxicity and good photostability were used as in vivo light conversion material for direct utilization of UV light in the solar energy. Copyright © 2018 Elsevier B.V. All rights reserved.

Solid-phase synthesis of graphene quantum dots from the food additive citric acid under microwave irradiation and their use in live-cell imaging.

PubMed

Zhuang, Qianfen; Wang, Yong; Ni, Yongnian

2016-05-01

The work demonstrated that solid citric acid, one of the most common food additives, can be converted to graphene quantum dots (GQDs) under microwave heating. The as-prepared GQDs were further characterized by various analytical techniques like transmission electron microscopy, atomic force microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, fluorescence and UV-visible spectroscopy. Cytotoxicity of the GQDs was evaluated using HeLa cells. The result showed that the GQDs almost did not exhibit cytotoxicity at concentrations as high as 1000 µg mL(-1). In addition, it was found that the GQDs showed good solubility, excellent photostability, and excitation-dependent multicolor photoluminescence. Subsequently, the multicolor GQDs were successfully used as a fluorescence light-up probe for live-cell imaging. Copyright © 2015 John Wiley & Sons, Ltd.

Contamination detection NDE for cleaning process inspection

NASA Technical Reports Server (NTRS)

Marinelli, W. J.; Dicristina, V.; Sonnenfroh, D.; Blair, D.

1995-01-01

In the joining of multilayer materials, and in welding, the cleanliness of the joining surface may play a large role in the quality of the resulting bond. No non-intrusive techniques are currently available for the rapid measurement of contamination on large or irregularly shaped structures prior to the joining process. An innovative technique for the measurement of contaminant levels in these structures using laser based imaging is presented. The approach uses an ultraviolet excimer laser to illuminate large and/or irregular surface areas. The UV light induces fluorescence and is scattered from the contaminants. The illuminated area is viewed by an image-intensified CCD (charge coupled device) camera interfaced to a PC-based computer. The camera measures the fluorescence and/or scattering from the contaminants for comparison with established standards. Single shot measurements of contamination levels are possible. Hence, the technique may be used for on-line NDE testing during manufacturing processes.

Reliability of a visual scoring system with fluorescent tracers to assess dermal pesticide exposure.

PubMed

Aragon, Aurora; Blanco, Luis; Lopez, Lylliam; Liden, Carola; Nise, Gun; Wesseling, Catharina

2004-10-01

We modified Fenske's semi-quantitative 'visual scoring system' of fluorescent tracer deposited on the skin of pesticide applicators and evaluated its reproducibility in the Nicaraguan setting. The body surface of 33 farmers, divided into 31 segments, was videotaped in the field after spraying with a pesticide solution containing a fluorescent tracer. A portable UV lamp was used for illumination in a foldaway dark room. The videos of five farmers were randomly selected. The scoring was based on a matrix with extension of fluorescent patterns (scale 0-5) on the ordinate and intensity (scale 0-5) on the abscissa, with the product of these two ranks as the final score for each body segment (0-25). Five medical students rated and evaluated the quality of 155 video images having undergone 4 h of training. Cronbach alpha coefficients and two-way random effects intraclass correlation coefficients (ICC) with absolute agreement were computed to assess inter-rater reliability. Consistency was high (Cronbach alpha = 0.96), but the scores differed substantially between raters. The overall ICC was satisfactory [0.75; 95% confidence interval (CI) = 0.62-0.83], but it was lower for intensity (0.54; 95% CI = 0.40-0.66) and higher for extension (0.80; 95% CI = 0.71-0.86). ICCs were lowest for images with low scores and evaluated as low quality, and highest for images with high scores and high quality. Inter-rater reliability coefficients indicate repeatability of the scoring system. However, field conditions for recording fluorescence should be improved to achieve higher quality images, and training should emphasize a better mechanism for the reading of body areas with low contamination.

Nonlethal screening of bat-wing skin with the use of ultraviolet fluorescence to detect lesions indicative of white-nose syndrome.

PubMed

Turner, Gregory G; Meteyer, Carol Uphoff; Barton, Hazel; Gumbs, John F; Reeder, DeeAnn M; Overton, Barrie; Bandouchova, Hana; Bartonička, Tomáš; Martínková, Natália; Pikula, Jiri; Zukal, Jan; Blehert, David S

2014-07-01

Definitive diagnosis of the bat disease white-nose syndrome (WNS) requires histologic analysis to identify the cutaneous erosions caused by the fungal pathogen Pseudogymnoascus [formerly Geomyces] destructans (Pd). Gross visual inspection does not distinguish bats with or without WNS, and no nonlethal, on-site, preliminary screening methods are available for WNS in bats. We demonstrate that long-wave ultraviolet (UV) light (wavelength 366-385 nm) elicits a distinct orange-yellow fluorescence in bat-wing membranes (skin) that corresponds directly with the fungal cupping erosions in histologic sections of skin that are the current gold standard for diagnosis of WNS. Between March 2009 and April 2012, wing membranes from 168 North American bat carcasses submitted to the US Geological Survey National Wildlife Health Center were examined with the use of both UV light and histology. Comparison of these techniques showed that 98.8% of the bats with foci of orange-yellow wing fluorescence (n=80) were WNS-positive based on histologic diagnosis; bat wings that did not fluoresce under UV light (n=88) were all histologically negative for WNS lesions. Punch biopsy samples as small as 3 mm taken from areas of wing with UV fluorescence were effective for identifying lesions diagnostic for WNS by histopathology. In a nonlethal biopsy-based study of 62 bats sampled (4-mm diameter) in hibernacula of the Czech Republic during 2012, 95.5% of fluorescent (n=22) and 100% of nonfluorescent (n=40) wing samples were confirmed by histopathology to be WNS positive and negative, respectively. This evidence supports use of long-wave UV light as a nonlethal and field-applicable method to screen bats for lesions indicative of WNS. Further, UV fluorescence can be used to guide targeted, nonlethal biopsy sampling for follow-up molecular testing, fungal culture analysis, and histologic confirmation of WNS.

Nonlethal screening of bat-wing skin with the use of ultraviolet fluorescence to detect lesions indicative of white-nose syndrome

USGS Publications Warehouse

Turner, Gregory G.; Meteyer, Carol U.; Barton, Hazel; Gumbs, John F.; Reeder, DeeAnn M.; Overton, Barrie; Bandouchova, Hana; Bartonička, Tomáš; Martínková, Natália; Pikula, Jiri; Zukal, Jan; Blehert, David S.

2014-01-01

Definitive diagnosis of the bat disease white-nose syndrome (WNS) requires histologic analysis to identify the cutaneous erosions caused by the fungal pathogen Pseudogymnoascus [formerly Geomyces] destructans (Pd). Gross visual inspection does not distinguish bats with or without WNS, and no nonlethal, on-site, preliminary screening methods are available for WNS in bats. We demonstrate that long-wave ultraviolet (UV) light (wavelength 368–385 nm) elicits a distinct orange–yellow fluorescence in bat-wing membranes (skin) that corresponds directly with the fungal cupping erosions in histologic sections of skin that are the current gold standard for diagnosis of WNS. Between March 2009 and April 2012, wing membranes from 168 North American bat carcasses submitted to the U.S. Geological Survey National Wildlife Health Center were examined with the use of both UV light and histology. Comparison of these techniques showed that 98.8% of the bats with foci of orange–yellow wing fluorescence (n = 80) were WNS-positive based on histologic diagnosis; bat wings that did not fluoresce under UV light (n = 88) were all histologically negative for WNS lesions. Punch biopsy samples as small as 3 mm taken from areas of wing with UV fluorescence were effective for identifying lesions diagnostic for WNS by histopathology. In a nonlethal biopsy-based study of 62 bats sampled (4-mm diameter) in hibernacula of the Czech Republic during 2012, 95.5% of fluorescent (n = 22) and 100% of nonfluorescent (n = 40) wing samples were confirmed by histopathology to be WNS positive and negative, respectively. This evidence supports use of long-wave UV light as a nonlethal and field-applicable method to screen bats for lesions indicative of WNS. Further, UV fluorescence can be used to guide targeted, nonlethal biopsy sampling for follow-up molecular testing, fungal culture analysis, and histologic confirmation of WNS.

Effects of iron on optical properties of dissolved organic matter.

PubMed

Poulin, Brett A; Ryan, Joseph N; Aiken, George R

2014-09-02

Iron is a source of interference in the spectroscopic analysis of dissolved organic matter (DOM); however, its effects on commonly employed ultraviolet and visible (UV-vis) light adsorption and fluorescence measurements are poorly defined. Here, we describe the effects of iron(II) and iron(III) on the UV-vis absorption and fluorescence of solutions containing two DOM fractions and two surface water samples. In each case, regardless of DOM composition, UV-vis absorption increased linearly with increasing iron(III). Correction factors were derived using iron(III) absorption coefficients determined at wavelengths commonly used to characterize DOM. Iron(III) addition increased specific UV absorbances (SUVA) and decreased the absorption ratios (E2:E3) and spectral slope ratios (SR) of DOM samples. Both iron(II) and iron(III) quenched DOM fluorescence at pH 6.7. The degree and region of fluorescence quenching varied with the iron:DOC concentration ratio, DOM composition, and pH. Regions of the fluorescence spectra associated with greater DOM conjugation were more susceptible to iron quenching, and DOM fluorescence indices were sensitive to the presence of both forms of iron. Analyses of the excitation-emission matrices using a 7- and 13-component parallel factor analysis (PARAFAC) model showed low PARAFAC sensitivity to iron addition.

Towards label-free evaluation of oxidative stress in human skin exposed to sun filters (Conference Presentation)

NASA Astrophysics Data System (ADS)

Osseiran, Sam; Wang, Hequn; Suita, Yusuke; Roider, Elisabeth; Fisher, David E.; Evans, Conor L.

2016-02-01

Skin cancer, including basal cell carcinoma, squamous cell carcinoma, and melanoma, is the most common form of cancer in North America. Paradoxically, skin cancer incidence is steadily on the rise even despite the growing use of sunscreens over the past decades. One potential explanation for this discrepancy involves the sun filters in sunscreen, which are responsible for blocking harmful ultraviolet radiation. It is proposed that these agents may produce reactive oxygen species (ROS) at the site of application, thereby generating oxidative stress in skin that gives rise to genetic mutations, which may explain the rising incidence of skin cancer. To test this hypothesis, ex vivo human skin was treated with five common chemical sun filters (avobenzone, octocrylene, homosalate, octisalate, and oxybenzone) as well as two physical sun filters (zinc oxide compounds), both with and without UV irradiation. To non-invasively evaluate oxidative stress, two-photon excitation fluorescence (2PEF) and fluorescence lifetime imaging microscopy (FLIM) of the skin samples were used to monitor levels of NADH and FAD, two key cofactors in cellular redox metabolism. The relative redox state of the skin was assessed based on the fluorescence intensities and lifetimes of these endogenous cofactors. While the sun filters were indeed shown to have a protective effect from UV radiation, it was observed that they also generate oxidative stress in skin, even in the absence of UV light. These results suggest that sun filter induced ROS production requires more careful study, especially in how these reactive species impact the rise of skin cancer.

Fluorescence quantum yield of carbon dioxide for quantitative UV laser-induced fluorescence in high-pressure flames

NASA Astrophysics Data System (ADS)

Lee, T.; Bessler, W. G.; Yoo, J.; Schulz, C.; Jeffries, J. B.; Hanson, R. K.

2008-11-01

The fluorescence quantum yield for ultraviolet laser-induced fluorescence of CO2 is determined for selected excitation wavelengths in the range 215-250 nm. Wavelength-resolved laser-induced fluorescence (LIF) spectra of CO2, NO, and O2 are measured in the burned gases of a laminar CH4/air flame ( φ=0.9 and 1.1) at 20 bar with additional NO seeded into the flow. The fluorescence spectra are fit to determine the relative contribution of the three species to infer an estimate of fluorescence quantum yield for CO2 that ranges from 2-8×10-6 depending on temperature and excitation wavelength with an estimated uncertainty of ±0.5×10-6. The CO2 fluorescence signal increases linearly with gas pressure for flames with constant CO2 mole fraction for the 10 to 60 bar range, indicating that collisional quenching is not an important contributor to the CO2 fluorescence quantum yield. Spectral simulation calculations are used to choose two wavelengths for excitation of CO2, 239.34 and 242.14 nm, which minimize interference from LIF of NO and O2. Quantitative LIF images of CO2 are demonstrated using these two excitation wavelengths and the measured fluorescence quantum yield.

Porphyrin synthesized from cashew nut shell liquid as part of a novel superparamagnetic fluorescence nanosystem

NASA Astrophysics Data System (ADS)

Clemente, C. S.; Ribeiro, V. G. P.; Sousa, J. E. A.; Maia, F. J. N.; Barreto, A. C. H.; Andrade, N. F.; Denardin, J. C.; Mele, G.; Carbone, L.; Mazzetto, S. E.; Fechine, P. B. A.

2013-06-01

Magnetic Fe3O4 nanoparticles with average size approximately 11 nm were first oleic acid coated to interact with the meso-porphyrin derivative from CNSL. This procedure produced a novel superparamagnetic fluorescent nanosystem (SFN) linked by van der Waals interactions. This system was characterized by transmission electron microscope, infrared spectroscopy, thermogravimetric analysis, magnetic measurements, UV-Vis absorption, and fluorescence emission measurements. These results showed that SFN has good thermal stability, excellent magnetization, and nanosized dimensions ( 13 nm). It exhibited emission peaks at 668 and 725 nm with a maximum emission at 467 nm of excitation wavelength. The type of interaction between porphyrin and magnetic nanoparticles allowed to obtain a material with interesting optical properties which might be used as an imaging agent for contrast in cells as well as heterogeneous photocatalysis.

Investigation of holmium-doped zirconium oxide ceramic phosphor as an ultraviolet wavelength-discriminating laser beam viewer

NASA Astrophysics Data System (ADS)

Yamanoi, Kohei; Hori, Tatsuhiro; Minami, Yuki; Empizo, Melvin John F.; Luong, Mui Viet; Shiro, Atsushi; Watanabe, Jun; Iwano, Keisuke; Iwasa, Yuki; Cadatal-Raduban, Marilou; Gabayno, Jacque Lynn; Shimizu, Toshihiko; Sarukura, Nobuhiko; Norimatsu, Takayoshi

2018-01-01

We report the fluorescence spectra of ZrO2 and trivalent Ho-doped ZrO2 ceramics under ultraviolet (UV) excitation at 213, 266, and 355 nm wavelengths. The Ho3+-doped ZrO2 ceramics exhibited varying fluorescence color tones depending on the excitation wavelength used. The different color tones match the fluorescence spectrum characteristics at each excitation wavelength. Our results demonstrate that Ho3+-doped ZrO2 ceramics can discriminate between UV light, specifically the third, fourth, and fifth harmonics of a Nd:YAG laser. It can potentially be used for developing UV laser beam viewers to aid laser alignment.

Synthesis, characterization, crystal structure and DNA-binding studies of transition metal hydrazone complexes

NASA Astrophysics Data System (ADS)

Kanchanadevi, S.; Parveen, S.; Mahalingam, V.

2018-04-01

Three new complexes containing salicylaldazine (HL) ligand were synthesised by reacting suitable precursor complex [MCl2(PPh3)2] with the ligand (where M = Cu(II) or Ni(II) or Co(II)). The new complexes were characterised by various spectral studies such as IR, UV-Vis,1H NMR,EPR,fluorescence and elemental analyses. The binding modes of the complexes with HS-DNA have been studied by UV-Vis absorption titration. Binding of the complexes with bovine serum albumin (BSA) protein has been investigated using UV-visible, fluorescence and synchronous fluorescence spectroscopic methods. Redox behaviour of the complexes has been investigated by cyclic voltammetry.

Enzyme-dependent fluorescence recovery of NADH after photobleaching to assess dehydrogenase activity of isolated perfused hearts

NASA Astrophysics Data System (ADS)

Moreno, Angel; Kuzmiak-Glancy, Sarah; Jaimes, Rafael; Kay, Matthew W.

2017-03-01

Reduction of NAD+ by dehydrogenase enzymes to form NADH is a key component of cellular metabolism. In cellular preparations and isolated mitochondria suspensions, enzyme-dependent fluorescence recovery after photobleaching (ED-FRAP) of NADH has been shown to be an effective approach for measuring the rate of NADH production to assess dehydrogenase enzyme activity. Our objective was to demonstrate how dehydrogenase activity could be assessed within the myocardium of perfused hearts using NADH ED-FRAP. This was accomplished using a combination of high intensity UV pulses to photobleach epicardial NADH. Replenishment of epicardial NADH fluorescence was then imaged using low intensity UV illumination. NADH ED-FRAP parameters were optimized to deliver 23.8 mJ of photobleaching light energy at a pulse width of 6 msec and a duty cycle of 50%. These parameters provided repeatable measurements of NADH production rate during multiple metabolic perturbations, including changes in perfusate temperature, electromechanical uncoupling, and acute ischemia/reperfusion injury. NADH production rate was significantly higher in every perturbation where the energy demand was either higher or uncompromised. We also found that NADH production rate remained significantly impaired after 10 min of reperfusion after global ischemia. Overall, our results indicate that myocardial NADH ED-FRAP is a useful optical non-destructive approach for assessing dehydrogenase activity.

Detecting proteins in highly autofluorescent cells using quantum dot antibody conjugates.

PubMed

Orcutt, Karen M; Ren, Shanshan; Gundersen, Kjell

2009-01-01

We have applied quantum dot (Qdot) antibody conjugates as a biomolecular probe for cellular proteins important in biogeochemical cycling in the sea. Conventional immunological methods have been hampered by the strong autofluorescence found in cyanobacteria cells. Qdot conjugates provide an ideal alternative for studies that require long-term imaging of cells such as detection of low abundance cellular antigens by fluorescence microscopy. The advantage of Qdot labeled probes over conventional immunological methods is the photostability of the probe. Phycoerythrin bleaches in cyanobacterial cells under prolonged UV or blue light excitation, which means that the semiconducting nanocrystal probe, the Qdot, can yield a strong fluorescent signal without interference from cellular pigments.

Digital image processing of bone - Problems and potentials

NASA Technical Reports Server (NTRS)

Morey, E. R.; Wronski, T. J.

1980-01-01

The development of a digital image processing system for bone histomorphometry and fluorescent marker monitoring is discussed. The system in question is capable of making measurements of UV or light microscope features on a video screen with either video or computer-generated images, and comprises a microscope, low-light-level video camera, video digitizer and display terminal, color monitor, and PDP 11/34 computer. Capabilities demonstrated in the analysis of an undecalcified rat tibia include the measurement of perimeter and total bone area, and the generation of microscope images, false color images, digitized images and contoured images for further analysis. Software development will be based on an existing software library, specifically the mini-VICAR system developed at JPL. It is noted that the potentials of the system in terms of speed and reliability far exceed any problems associated with hardware and software development.

Fluorescent minerals--A potential source of UV protection and visible light for the growth of green algae and cyanobacteria in extreme cosmic environments.

PubMed

Omairi, Tareq; Wainwright, Milton

2015-07-01

We propose that green algae (Chlorella variabilis and Dunaliella tertiolecta) and cyanobacteria (Synechococcus elongatus and Nostoc commune) can grow inside fluorescent rock minerals which convert damaging UV light to visible light, thereby allowing these organisms to survive and thrive in UV-rich environments without (or with limited) visible light, which would otherwise be inimical to them. The four microorganisms were incubated inside fluorescent rocks composed of fluorite, calcite and pyrite. The resultant growth was then measured following exposure to UV radiation, with the use of optical density and measurement of chlorophyll concentration. Results show that the microorganisms were shielded from harmful UV in these semi-transparent rocks, while at the same time benefiting from the fact that the minerals converted UV to visible light; this have been shown by a statistically significant increase in their growth, which although lower than when the cells were incubated in sunlight, was significantly higher than in controls incubated in the dark. Copyright © 2015 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.

CHAMP - Camera, Handlens, and Microscope Probe

NASA Technical Reports Server (NTRS)

Mungas, G. S.; Beegle, L. W.; Boynton, J.; Sepulveda, C. A.; Balzer, M. A.; Sobel, H. R.; Fisher, T. A.; Deans, M.; Lee, P.

2005-01-01

CHAMP (Camera, Handlens And Microscope Probe) is a novel field microscope capable of color imaging with continuously variable spatial resolution from infinity imaging down to diffraction-limited microscopy (3 micron/pixel). As an arm-mounted imager, CHAMP supports stereo-imaging with variable baselines, can continuously image targets at an increasing magnification during an arm approach, can provide precision range-finding estimates to targets, and can accommodate microscopic imaging of rough surfaces through a image filtering process called z-stacking. Currently designed with a filter wheel with 4 different filters, so that color and black and white images can be obtained over the entire Field-of-View, future designs will increase the number of filter positions to include 8 different filters. Finally, CHAMP incorporates controlled white and UV illumination so that images can be obtained regardless of sun position, and any potential fluorescent species can be identified so the most astrobiologically interesting samples can be identified.

Characterization of aeroallergen of Texas panhandle using scanning and fluorescence microscopy

NASA Astrophysics Data System (ADS)

Ghosh, Nabarun; Whiteside, Mandy; Ridner, Chris; Celik, Yasemin; Saadeh, C.; Bennert, Jeff

2010-06-01

Aeroallergens cause serious allergic and asthmatic reactions. Characterizing the aeroallergen provides information regarding the onset, duration, and severity of the pollen season that clinicians use to guide allergen selection for skin testing and treatment. Fluorescence Microscopy has useful approaches to understand the structure and function of the microscopic objects. Prepared slides from the pollen were observed under an Olympus BX40 microscope equipped with FITC and TRITC fluorescent filters, a mercury lamp source, an Olympus DP-70 digital camera connected to the computer with Image Pro 6.0 software. Aeroallergens were viewed, recorded and analyzed with DP Manager using the Image Pro 6.0 software. Photographs were taken at bright field, the fluorescein-isothiocyanate (FITC) filter, and the tetramethylrhodamine (TRITC) filter settings at 40X. A high pressure mercury lamp or UV source was used to excite the storage molecules or proteins which exhibited autofluorescence. The FITC filter reveals the green fluorescent proteins (GFP and EGFP), and the TRITC filter for red fluorescent proteins (DsRed). SEM proved to be useful for observing ultra-structural details like pores, colpi, sulci and ornamentations on the pollen surface. Samples were examined with an SEM (TM-1000) after gold coating and Critical Point Drying. Pollen grains were measured using the TM-1000 imaging software that revealed the specific information on the size of colpi or sulci and the distance between the micro-structures. This information can be used for classification and circumscription in Angiosperm taxonomy. Data were correlated clinical studies established at Allergy A.R.T.S. Clinical Research Laboratory.

Near-Infrared Emitting Squaraine Dyes with High 2PA Cross Sections for Multiphoton Fluorescence Imaging

PubMed Central

Ahn, Hyo-Yang; Yao, Sheng; Wang, Xuhua; Belfield, Kevin D.

2012-01-01

Designed to achieve high two-photon absorptivity, new near infrared (NIR) emitting squaraine dyes, (E)-2-(1-(2-(2-methoxyethoxy)ethyl)-5-(3,4,5-trimethoxystyryl)-1H-pyrrol-2-yl)-4-(1-(2-(2-methoxyethoxy)ethyl)-5-(3,4,5-trimethoxystyryl)-2H-pyrrolium-2-ylidene)-3-oxocyclobut-1-enolate (1) and (Z)-2-(4-(dibutylamino)-2-hydroxyphenyl)-4-(4-(dibutyliminio)-2-hydroxycyclohexa-2,5-dienylidene)-3-oxocyclobut-1-enolate (2) were synthesized and characterized. Their linear photophysical properties were investigated via UV-visible absorption spectroscopy and fluorescence spectroscopy in various solvents, while their nonlinear photophysical properties were investigated using a combination of two-photon induced fluorescence and open aperture z-scan methods. Squaraine 1 exhibited a high two-photon absorption (2PA) cross section (δ2PA), ~ 20,000 GM at 800 nm, and high photostability with the photochemical decomposition quantum yield one order of magnitude lower than Cy 5, a commercially available pentamethine cyanine NIR dye. The cytotoxicity of the squaraine dyes were evaluated in HCT 116 and COS 7 cell lines to assess the potential of these probes for biomedical imaging. The viability of both cell lines was maintained above 80% at dye concentrations up to 30 μM, indicating good biocompatibility of the probes. Finally, one-photon fluorescence microscopy (1PFM) and two-photon fluorescence microscopy (2PFM) imaging was accomplished after incubation of micelle-encapsulated squaraine probes with HCT 116 and COS 7 cells, demonstrating their potential in 2PFM bioimaging. PMID:22591003

The role of ultraviolet-adaptation of a marine diatom in photoenhanced toxicity of acridine.

PubMed

Wiegman, Saskia; Barranguet, Christiane; Spijkerman, Elly; Kraak, Michiel Harm Steven; Admiraal, Wim

2003-03-01

Cultures of the marine diatom Phaeodactylum tricornutum were grown under laboratory light with a different fraction of ultraviolet radiation (UV) to study the potential role of photoadaptation in determining the sensitivity to photoenhanced toxicity of acridine. In short-term experiments, a higher acridine concentration was needed to inhibit the photosynthetic electron flux, monitored with chlorophyll a fluorescence, in algae exposed to fluorescent light (low UV) than to mercury light (high UV), consistent with the expected role of UV. The two types of light in long-term exposures led to changes in the pigment composition and photosystem I (PS I) to photosystem II (PS II) stoichiometry to optimize the utilization of fluorescent and mercury light. Despite the adaptation of the photosynthetic apparatus to a small fraction of UV, long-term exposure to mercury light did show a constant sensitivity of the photosynthetic efficiency of P. tricornutum to the phototoxic acridine. It is concluded that the prime receptor of photoenhanced toxicity may be unrelated to the photosynthetic machinery.

Time-gated FLIM microscope for corneal metabolic imaging

NASA Astrophysics Data System (ADS)

Silva, Susana F.; Batista, Ana; Domingues, José Paulo; Quadrado, Maria João.; Morgado, António Miguel

2016-03-01

Detecting corneal cells metabolic alterations may prove a valuable tool in the early diagnosis of corneal diseases. Nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) are autofluorescent metabolic co-factors that allow the assessment of metabolic changes through non-invasive optical methods. These co-factors exhibit double-exponential fluorescence decays, with well-separated short and lifetime components, which are related to their protein-bound and free-states. Corneal metabolism can be assessed by measuring the relative contributions of these two components. For that purpose, we have developed a wide-field time-gated fluorescence lifetime microscope based on structured illumination and one-photon excitation to record FAD lifetime images from corneas. NADH imaging was not considered as its UV excitation peak is regarded as not safe for in vivo measurements. The microscope relies on a pulsed blue diode laser (λ=443 nm) as excitation source, an ultra-high speed gated image intensifier coupled to a CCD camera to acquire fluorescence signals and a Digital Micromirror Device (DMD) to implement the Structured Illumination technique. The system has a lateral resolution better than 2.4 μm, a field of view of 160 per 120 μm and an optical sectioning of 6.91 +/- 0.45 μm when used with a 40x, 0.75 NA, Water Immersion Objective. With this setup we were able to measure FAD contributions from ex-vivo chicken corneas collected from a local slaughterhouse..

Development of novel small molecules for imaging and drug release

NASA Astrophysics Data System (ADS)

Cao, Yanting

Small organic molecules, including small molecule based fluorescent probes, small molecule based drugs or prodrugs, and smart multifunctional fluorescent drug delivery systems play important roles in biological research, drug discovery, and clinical practices. Despite the significant progress made in these fields, the development of novel and diverse small molecules is needed to meet various demands for research and clinical applications. My Ph.D study focuses on the development of novel functional molecules for recognition, imaging and drug release. In the first part, a turn-on fluorescent probe is developed for the detection of intracellular adenosine-5'-triphosphate (ATP) levels based on multiplexing recognitions. Considering the unique and complicated structure of ATP molecules, a fluorescent probe has been implemented with improved sensitivity and selectivity due to two synergistic binding recognitions by incorporating of 2, 2'-dipicolylamine (Dpa)-Zn(II) for targeting of phospho anions and phenylboronic acid group for cis-diol moiety. The novel probe is able to detect intracellular ATP levels in SH-SY5Y cells. Meanwhile, the advantages of multiplexing recognition design concept have been demonstrated using two control molecules. In the second part, a prodrug system is developed to deliver multiple drugs within one small molecule entity. The prodrug is designed by using 1-(2-nitrophenyl)ethyl (NPE) as phototrigger, and biphenol biquaternary ammonium as the prodrug. With controlled photo activation, both DNA cross-linking agents mechlorethamine and o-quinone methide are delivered and released at the preferred site, leading to efficient DNA cross-links formation and cell death. The prodrug shows negligible cytotoxicity towards normal skin cells (Hekn cells) with and without UV activation, but displays potent activity towards cancer cells (HeLa cells) upon UV activation. The multiple drug release system may hold a great potential for practical application. In the last part, a new photo-initiated fluorescent anticancer prodrug for DNA alkylating agent mechlorethamine releasing and monitoring has been developed. The theranostic prodrug consists a photolabile NPE group, an inactive form of mechlorethamine and a nonfluorescent coumarin in one small molecule. It is demonstrated that the prodrug shows negligible cytotoxicity towards normal skin cells (Hekn cells) with and without UV activation, while the original parent drug mechlorethamine can be photocontrol-released and induces effective DNA cross-linking activity. Importantly, the drug release progress can be conveniently monitored by the 'off-on' fluorescence enhancement in cells. Moreover, the selective prodrug is not only cell permeable but also nuclear permeable. Therefore, the prodrug serves as a promising drug delivery system for spatiotemporal control release and monitoring of an anticancer drug to obtain the optimal treatment efficacy.

Organic-inorganic interface-induced multi-fluorescence of MgO nanocrystal clusters and their applications in cellular imaging.

PubMed

Xie, Shuifen; Bao, Shixiong; Ouyang, Junjie; Zhou, Xi; Kuang, Qin; Xie, Zhaoxiong; Zheng, Lansun

2014-04-25

Surface functionalization of inorganic nanomaterials through chemical binding of organic ligands on the surface unsaturated atoms, forming unique organic-inorganic interfaces, is a powerful approach for creating special functions for inorganic nanomaterials. Herein, we report the synthesis of hierarchical MgO nanocrystal clusters (NCs) with an organic-inorganic interface induced multi-fluorescence and their application as new alternative labels for cellular imaging. The synthetic method was established by a dissolution and regrowth process with the assistance of carboxylic acid, in which the as-prepared MgO NCs were modified with carboxylic groups at the coordinatively unsaturated atoms of the surface. By introducing acetic acid to partially replace oleic acid in the reaction, the optical absorption of the produced MgO NCs was progressively engineered from the UV to the visible region. Importantly, with wider and continuous absorption profile, those MgO NCs presented bright and tunable multicolor emissions from blue-violet to green and yellow, with the highest absolute quantum yield up to (33±1) %. The overlap for the energy levels of the inorganic-organic interface and low-coordinated states stimulated a unique fluorescence resonance energy transfer phenomenon. Considering the potential application in cellular imaging, such multi-fluorescent MgO NCs were further encapsulated with a silica shell to improve the water solubility and stability. As expected, the as-formed MgO@SiO2 NCs possessed great biocompatibility and high performance in cellular imaging. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Smartphone-Based Dual-Modality Imaging System for Quantitative Detection of Color or Fluorescent Lateral Flow Immunochromatographic Strips

NASA Astrophysics Data System (ADS)

Hou, Yafei; Wang, Kan; Xiao, Kun; Qin, Weijian; Lu, Wenting; Tao, Wei; Cui, Daxiang

2017-04-01

Nowadays, lateral flow immunochromatographic assays are increasingly popular as a diagnostic tool for point-of-care (POC) test based on their simplicity, specificity, and sensitivity. Hence, quantitative detection and pluralistic popular application are urgently needed in medical examination. In this study, a smartphone-based dual-modality imaging system was developed for quantitative detection of color or fluorescent lateral flow test strips, which can be operated anywhere at any time. In this system, the white and ultra-violet (UV) light of optical device was designed, which was tunable with different strips, and the Sobel operator algorithm was used in the software, which could enhance the identification ability to recognize the test area from the background boundary information. Moreover, this technology based on extraction of the components from RGB format (red, green, and blue) of color strips or only red format of the fluorescent strips can obviously improve the high-signal intensity and sensitivity. Fifty samples were used to evaluate the accuracy of this system, and the ideal detection limit was calculated separately from detection of human chorionic gonadotropin (HCG) and carcinoembryonic antigen (CEA). The results indicated that smartphone-controlled dual-modality imaging system could provide various POC diagnoses, which becomes a potential technology for developing the next-generation of portable system in the near future.

Enchanced methods of hydrophilized CdSe quantum dots synthesis

NASA Astrophysics Data System (ADS)

Potapkin, D. V.; Zharkova, I. S.; Goryacheva, I. Y.

2015-03-01

Quantum dots are bright and stable fluorescence signal sources, but for most of applications they need an additional hydrophilization step. Unfortunately, most of existing approaches lead to QD's fluorescence quenching, so there is a need for additional enhancing of hydrophilized QD's brightness like UV irradiation, which can be used both on water insoluble QD's with oleic acid ligands (in toluene) and on hydrophilized QD's covered with UV-stable polymer (in aqueous solution). For synthesis of bright water-soluble fluorescent labels CdSe/CdS/ZnS colloidal quantum dots were covered with PAMAM dendrimer and irradiated with UV lamp in quartz cuvettes for 3 hours at the room temperature and then compared with control sample.

Applications of Light-Responsive Systems for Cancer Theranostics.

PubMed

Chen, Hongzhong; Zhao, Yanli

2018-06-27

Achieving controlled and targeted delivery of chemotherapeutic drugs and other therapeutic agents to tumor sites is challenging. Among many stimulus strategies, light as a mode of action shows various advantages such as high spatiotemporal selectivity, minimal invasiveness and easy operation. Thus, drug delivery systems (DDSs) have been designed with the incorporation of various functionalities responsive to light as an exogenous stimulus. Early development has focused on guiding chemotherapeutic drugs to designated location, followed by the utilization of UV irradiation for controlled drug release. Because of the disadvantages of UV light such as phototoxicity and limited tissue penetration depth, scientists have moved the research focus onto developing nanoparticle systems responsive to light in the visible region (400-700 nm), aiming to reduce the phototoxicity. In order to enhance the tissue penetration depth, near-infrared light triggered DDSs become increasingly important. In addition, light-based advanced systems for fluorescent and photoacoustic imaging, as well as photodynamic and photothermal therapy have also been reported. Herein, we highlight some of recent developments by applying light-responsive systems in cancer theranostics, including light activated drug release, photodynamic and photothermal therapy, and bioimaging techniques such as fluorescent and photoacoustic imaging. Future prospect of light-mediated cancer treatment is discussed at the end of the review. This Spotlights on Applications article aims to provide up-to-date information about the rapidly developing field of light-based cancer theranostics.

Detection of herbicide effects on pigment composition and PSII photochemistry in Helianthus annuus by Raman spectroscopy and chlorophyll a fluorescence

NASA Astrophysics Data System (ADS)

Vítek, Petr; Novotná, Kateřina; Hodaňová, Petra; Rapantová, Barbora; Klem, Karel

2017-01-01

The effects of herbicides from three mode-of-action groups - inhibitors of protoporphyrinogen oxidase (carfentrazone-ethyl), inhibitors of carotenoid biosynthesis (mesotrione, clomazone, and diflufenican), and inhibitors of acetolactate synthase (amidosulfuron) - were studied in sunflower plants (Helianthus annuus). Raman spectroscopy, chlorophyll fluorescence (ChlF) imaging, and UV screening of ChlF were combined to evaluate changes in pigment composition, photosystem II (PSII) photochemistry, and non-photochemical quenching in plant leaves 6 d after herbicide application. The Raman signals of phenolic compounds, carotenoids, and chlorophyll were evaluated and differences in their intensity ratios were observed. Strongly augmented relative content of phenolic compounds was observed in the case of amidosulfuron-treated plants, with a simultaneous decrease in the chlorophyll/carotenoid intensity ratio. The results were confirmed by in vivo measurement of flavonols using UV screening of ChlF. Herbicides from the group of carotenoid biosynthesis inhibitors significantly decreased both the maximum quantum efficiency of PSII and non-photochemical quenching as determined by ChlF. Resonance Raman imaging (mapping) data with high resolution (150,000-200,000 spectra) are presented, showing the distribution of carotenoids in H. annuus leaves treated by two of the herbicides acting as inhibitors of carotenoid biosynthesis (clomazone or diflufenican). Clear signs were observed that the treatment induced carotenoid depletion within sunflower leaves. The depletion spatial pattern registered differed depending on the type of herbicide applied.

Neuroendocrine Tumor-Targeted Upconversion Nanoparticle-Based Micelles for Simultaneous NIR-Controlled Combination Chemotherapy and Photodynamic Therapy, and Fluorescence Imaging.

PubMed

Chen, Guojun; Jaskula-Sztul, Renata; Esquibel, Corinne R; Lou, Irene; Zheng, Qifeng; Dammalapati, Ajitha; Harrison, April; Eliceiri, Kevin W; Tang, Weiping; Chen, Herbert; Gong, Shaoqin

2017-02-23

Although neuroendocrine tumors (NETs) are slow growing, they are frequently metastatic at the time of discovery and no longer amenable to curative surgery, emphasizing the need for the development of other treatments. In this study, multifunctional upconversion nanoparticle (UCNP)-based theranostic micelles are developed for NET-targeted and near-infrared (NIR)-controlled combination chemotherapy and photodynamic therapy (PDT), and bioimaging. The theranostic micelle is formed by individual UCNP functionalized with light-sensitive amphiphilic block copolymers poly(4,5-dimethoxy-2-nitrobenzyl methacrylate)-polyethylene glycol (PNBMA-PEG) and Rose Bengal (RB) photosensitizers. A hydrophobic anticancer drug, AB3, is loaded into the micelles. The NIR-activated UCNPs emit multiple luminescence bands, including UV, 540 nm, and 650 nm. The UV peaks overlap with the absorption peak of photocleavable hydrophobic PNBMA segments, triggering a rapid drug release due to the NIR-induced hydrophobic-to-hydrophilic transition of the micelle core and thus enabling NIR-controlled chemotherapy. RB molecules are activated via luminescence resonance energy transfer to generate 1 O 2 for NIR-induced PDT. Meanwhile, the 650 nm emission allows for efficient fluorescence imaging. KE108, a true pansomatostatin nonapeptide, as an NET-targeting ligand, drastically increases the tumoral uptake of the micelles. Intravenously injected AB3-loaded UCNP-based micelles conjugated with RB and KE108-enabling NET-targeted combination chemotherapy and PDT-induce the best antitumor efficacy.

Toxicity of Food-Grade TiO2 to Commensal Intestinal and Transient Food-Borne Bacteria: New Insights Using Nano-SIMS and Synchrotron UV Fluorescence Imaging.

PubMed

Radziwill-Bienkowska, Joanna M; Talbot, Pauline; Kamphuis, Jasper B J; Robert, Véronique; Cartier, Christel; Fourquaux, Isabelle; Lentzen, Esther; Audinot, Jean-Nicolas; Jamme, Frédéric; Réfrégiers, Matthieu; Bardowski, Jacek K; Langella, Philippe; Kowalczyk, Magdalena; Houdeau, Eric; Thomas, Muriel; Mercier-Bonin, Muriel

2018-01-01

Titanium dioxide (TiO 2 ) is commonly used as a food additive (E171 in the EU) for its whitening and opacifying properties. However, a risk of intestinal barrier disruption, including dysbiosis of the gut microbiota, is increasingly suspected because of the presence of a nano-sized fraction in this additive. We hypothesized that food-grade E171 and Aeroxyde P25 (identical to the NM-105 OECD reference nanomaterial in the European Union Joint Research Centre) interact with both commensal intestinal bacteria and transient food-borne bacteria under non-UV-irradiated conditions. Based on differences in their physicochemical properties, we expect a difference in their respective effects. To test these hypotheses, we chose a panel of eight Gram-positive/Gram-negative bacterial strains, isolated from different biotopes and belonging to the species Escherichia coli , Lactobacillus rhamnosus , Lactococcus lactis (subsp. lactis and cremoris ), Streptococcus thermophilus , and Lactobacillus sakei . Bacterial cells were exposed to food-grade E171 vs. P25 in vitro and the interactions were explored with innovative (nano)imaging methods. The ability of bacteria to trap TiO 2 was demonstrated using synchrotron UV fluorescence imaging with single cell resolution. Subsequent alterations in the growth profiles were shown, notably for the transient food-borne L. lactis and the commensal intestinal E. coli in contact with food-grade TiO 2 . However, for both species, the reduction in cell cultivability remained moderate, and the morphological and ultrastructural damages, observed with electron microscopy, were restricted to a small number of cells. E. coli exposed to food-grade TiO 2 showed some internalization of TiO 2 (7% of cells), observed with high-resolution nano-secondary ion mass spectrometry (Nano-SIMS) chemical imaging. Taken together, these data show that E171 may be trapped by commensal and transient food-borne bacteria within the gut. In return, it may induce some physiological alterations in the most sensitive species, with a putative impact on gut microbiota composition and functioning, especially after chronic exposure.

Toxicity of Food-Grade TiO2 to Commensal Intestinal and Transient Food-Borne Bacteria: New Insights Using Nano-SIMS and Synchrotron UV Fluorescence Imaging

PubMed Central

Radziwill-Bienkowska, Joanna M.; Talbot, Pauline; Kamphuis, Jasper B. J.; Robert, Véronique; Cartier, Christel; Fourquaux, Isabelle; Lentzen, Esther; Audinot, Jean-Nicolas; Jamme, Frédéric; Réfrégiers, Matthieu; Bardowski, Jacek K.; Langella, Philippe; Kowalczyk, Magdalena; Houdeau, Eric; Thomas, Muriel; Mercier-Bonin, Muriel

2018-01-01

Titanium dioxide (TiO2) is commonly used as a food additive (E171 in the EU) for its whitening and opacifying properties. However, a risk of intestinal barrier disruption, including dysbiosis of the gut microbiota, is increasingly suspected because of the presence of a nano-sized fraction in this additive. We hypothesized that food-grade E171 and Aeroxyde P25 (identical to the NM-105 OECD reference nanomaterial in the European Union Joint Research Centre) interact with both commensal intestinal bacteria and transient food-borne bacteria under non-UV-irradiated conditions. Based on differences in their physicochemical properties, we expect a difference in their respective effects. To test these hypotheses, we chose a panel of eight Gram-positive/Gram-negative bacterial strains, isolated from different biotopes and belonging to the species Escherichia coli, Lactobacillus rhamnosus, Lactococcus lactis (subsp. lactis and cremoris), Streptococcus thermophilus, and Lactobacillus sakei. Bacterial cells were exposed to food-grade E171 vs. P25 in vitro and the interactions were explored with innovative (nano)imaging methods. The ability of bacteria to trap TiO2 was demonstrated using synchrotron UV fluorescence imaging with single cell resolution. Subsequent alterations in the growth profiles were shown, notably for the transient food-borne L. lactis and the commensal intestinal E. coli in contact with food-grade TiO2. However, for both species, the reduction in cell cultivability remained moderate, and the morphological and ultrastructural damages, observed with electron microscopy, were restricted to a small number of cells. E. coli exposed to food-grade TiO2 showed some internalization of TiO2 (7% of cells), observed with high-resolution nano-secondary ion mass spectrometry (Nano-SIMS) chemical imaging. Taken together, these data show that E171 may be trapped by commensal and transient food-borne bacteria within the gut. In return, it may induce some physiological alterations in the most sensitive species, with a putative impact on gut microbiota composition and functioning, especially after chronic exposure. PMID:29740421

The influence of indoxyl sulfate and ammonium on the autofluorescence of human urine.

PubMed

Perinchery, Sandeep Menon; Kuzhiumparambil, Unnikrishnan; Vemulpad, Subramanyam; Goldys, Ewa M

2010-01-15

Despite biological variability the spectral characteristics of undiluted human urine show relatively low autofluorescence at short UV (250-300nm) excitation. However with dilution the fluorescence intensity remarkably increases. This paper examines the mechanisms behind this effect, by using excitation-emission matrices. Corrections for the inner filter effect were made for improved understanding of the spectral patterns. We focused on three major fluorophores (tryptophan, indoxyl sulfate and 5-hydroxyindole-3-acetate) that are excited at these wavelengths, and whose content in urine is strongly linked with various health conditions. Their fluorescence was studied both individually and in combinations. We also examined the effect of ammonium on the fluorescence of these major fluorophores individually and in combinations. Through these studies we have identified the leading effects that reduce the UV fluorescence, namely higher concentration of indoxyl sulfate producing the inner filter effect and concentration quenching and quenching of fluorophores by ammonium. This result will assist in broader utilisation of UV fluorescence in medical diagnostics.

Laser-Stimulated Fluorescence in Paleontology

PubMed Central

Kaye, Thomas G.; Falk, Amanda R.; Pittman, Michael; Sereno, Paul C.; Burnham, David A.; Gong, Enpu; Xu, Xing; Wang, Yinan

2015-01-01

Fluorescence using ultraviolet (UV) light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF) is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser’s ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck. PMID:26016843

Infrared fluorescence microscopy of stained tissues: principles and technic.

PubMed

Puchtler, H; Meloan, S N; Paschal, L D

1980-01-01

Infrared photomicrography was used extensively from 1927 to the 1940's, but received little attention during the last decades. However, studies of infrared fluorescence of stained sections could not be found in the accessible literature. Ramsley (1968) published quantitative data on infrared fluorescence of approximately 250 dyes bound to textile fibers. The intensity of infrared fluorescence of many dyes varied widely with the substrate. It was therefore deemed of interest to determine whether or not similar differences in infrared fluorescence may occur when dyes are bound to histochemically distinct tissue structures. Myofibrils and collagens stained with triarylmethane dyes were chosen as test objects. Kodak infrared cut-off filter No. 301 and Wratten filter #16 were used as exciter filters to remove infrared and UV-blue and the light of a xenon lamp. Wratten filter #70 and #89B were employed as barrier filters. Infrared radiation was recorded with Kodak Ektachrome infrared film. To facilitate correlation of infrared fluorescence patterns with visible images, tissues were photographed also with conventional color film. Stained myofibrils, e.g. in myoepithelium, smooth and striated muscle emitted strong infrared fluorescence; collagen showed little or no fluorescence. Barrier filter Wratten #70 permitted simultaneous demonstration of infrared fluorescence and of non-fluorescent structures and thus facilitated histopathological studies. Preliminary findings indicate decrease or loss of infrared fluorescence of stained muscle fibers in various lesions, e.g. myocardial infarction, Duchenne-type muscular dystrophy.

SHERLOC: An investigation for Mars 2020

NASA Astrophysics Data System (ADS)

Beegle, Luther; Bhartia, Rohit

2016-04-01

SHERLOC is a Deep UV (DUV) native fluorescence and resonance Raman spectrometer that was selected as part of the Mars 2020 payload. It is a robotic arm mounted instrument that utilizes a DUV laser to generate characteristic Raman and fluorescence photons from a targeted spot. The DUV laser is co-boresighted to a context imager and integrated into an autofocusing/scanning optical system that allows us to correlate spectral signatures to surface textures, morphology and visible features. Additionally, it has recently been augmented with an imaging system that is a built-to-print version of the MArs Hand Lens Imager (MAHLI) instrument on the Mars Science Laboratory (MSL). Through the use of an internal scanning mirror, autofocusing lens, and a depth of focus of ±500 μm, the 100 μm laser spot can be systematically scanned over a 7x7 mm area with a fine-scale spatial resolution on natural or abraded surfaces and boreholes to a depth of at least 13 mm, without further arm movement. Through the use of the context imager, SHERLOC's data products can be combined with observations made by other instruments on the Mars 2020 payload. By bringing to bear multiple scientific instruments on a single sample, our ability to assess the habitability of ancient environments and search for potential biosignatures preserved within the geologic record will be greatly enhanced, making possible the selection of high-priority samples for caching. The SHERLOC investigation combines two spectral phenomena, fluorescence and pre-resonance/resonance DUV Raman scattering. These spectral features are resolvable when a high-radiance, narrow line-width, laser source illuminates a sample. In fluorescence, the incident photons are absorbed and re-emitted at a longer wavelength. The difference between the excitation and emission wavelength is the difference between the excitation frequency and the lowest electronic state frequency that increases with increasing aromatic structure (i.e., number of aromatic rings). Typical fluorescence cross-sections are 104 greater than traditional Raman, enabling the detection of sub-picograms levels of aromatic organic compounds. SHERLOC's narrow-linewidth (3 GHz) DUV laser (248.6 nm) enables fluorescence-free Raman scattering for additional classification of aromatics and aliphatic organics and minerals. Raman scattering is inelastic scattering where the loss of energy from the excitation energy (40225 cm-1) and defines the vibrational energy of a bond with which it interacted. This enables classification of bonds such as C-H, CN, C=O, C=C, NHx, NOx, SOx, POx, ClOx, and OH. It should be noted that Raman shifts (cm-1) (i.e., peak position as energy loss from the excitation energy) are invariant to excitation wavelength. Thus peak positions in Raman databases (at 229, 248, 488, 532 and 785 nm) can be compared (c.f. http://rruff.info/). Fluorescence emission of organics extends from ~270 nm into visible wavelengths. On the other hand, mineral fluorescence emission stemming from crystalline defects and impurities is weak in the deep UV, and typically begins longward of 360 nm continuing into the NIR. Mineral fluorescence is very unlikely to be seen in samples found on Mars. The DUV fluorescence technique employed by SHERLOC is well-suited to the detection of organics on mineral surfaces.

Design and daytime performance of laser-induced fluorescence spectrum lidar for simultaneous detection of multiple components, dissolved organic matter, phycocyanin, and chlorophyll in river water.

PubMed

Saito, Yasunori; Kakuda, Kei; Yokoyama, Mizuho; Kubota, Tomoki; Tomida, Takayuki; Park, Ho-Dong

2016-08-20

In this work, we developed mobile laser-induced fluorescence spectrum (LIFS) lidar based on preliminary experiments on the excitation emission matrix of a water sample and a method for reducing solar background light using the synchronous detection technique. The combination of a UV short-pulse laser (355 nm, 6 ns) for fluorescence excitation with a 10-100 ns short-time synchronous detection using a gated image-intensified multi-channel CCD of the fluorescence made the LIFS lidar operation possible even in daytime. The LIFS lidar with this construction demonstrated the potential of natural river/lake water quality monitoring at the Tenryu River/Lake Suwa. Three main components in the fluorescence data of the water, dissolved organic matter, phycocyanin, and chlorophyll, were extracted by spectral analysis using the standard spectral functions of these components. Their concentrations were estimated by adapting experimentally calibrated data. Results of long-term field observations using our LIFS lidar from 2010 to 2012 show the necessity of simultaneous multi-component detection to understand the natural water environment.

Effect of UV irradiation on the apoptosis and necrosis of Jurkat cells using UV LEDs

NASA Astrophysics Data System (ADS)

Inada, Shunko A.; Amano, Hiroshi; Akasaki, Isamu; Morita, Akimichi; Kobayashi, Keiko

2009-02-01

Phototherapy is a very effective method for treating most of the incurable skin diseases. A fluorescent light bulb is used as a conventional UV light source for this type of therapy. However, infrared radiation from the light source sometimes causes serious problems on patient's health. In addition, the normal part of the skin is irradiated when a large fluorescent light bulb is used. Moreover, a conventional UV irradiation system is heavy and has a short lifetime and a high electrical power consumption. Therefore, a new UV light source for solving the problems of phototherapy is required. To realize low-power-consumption, lightweight and long-lifetime systems, group III nitride-based UV-A1 light-emitting diodes (LEDs) were investigated. We examined the UV LED irradiation of Jurkat cell, which is a tumor cell and more sensitive to UV light than a healthy cell. The numbers of apoptotic and necrotic cells were confirmed to be the same using a UV LED and a conventional lamp system. The UV LED showed the possibility of realizing a new UV light source for phototherapy.

Preparation of multifunctional PEG-graft-Halloysite Nanotubes for Controlled Drug Release, Tumor Cell Targeting, and Bio-imaging.

PubMed

Yamina, Ait Mehdi; Fizir, Meriem; Itatahine, Asma; He, Hua; Dramou, Pierre

2018-06-19

With the intent of enhancing the loading capacity and controlled the release of a low-water soluble drug (quercetin), 6 ar ms (Poly-Ethylene-Glycol)-amine was grafted in the external surface of halloysite nanotubes. The grafted halloysite nanotubes (HNTs-g-PEG) were decorated with carbon quantum dots for additive fluorescents properties. Conjointly, biotin was conjugated to PEG's free amine groups for precise targeting of tumor tissue and higher cellular uptake. The obtained nanoparticles were characterized by the FTIR, TEM, XRD, zeta potential and TGA analysis. The photoluminescence (PL) properties were investigated by firstly, observing under UV-light at 365 nm; then, the fluorescence spectra of modified HNTs at different levels of preparation were obtained and showed a suitable blue fluorescence. Furthermore, the fluorescent properties were demonstrated by the optical image of HNTs-g-PEG-CDs-Biotin obtained from the confocal microscope, which could be interesting for both in vitro and in vivo imaging. Besides, the prepared NPs showed a superior loading capacity of Que (278.36 mg/g) at optimal adsorption conditions comparing to pristine HNTs. The in vitro drug release from these NPs was relatively sustained and pH sensitive. The incubation of the prepared HNTs-g-PEG-CDs-Biotin NPs with HeLa cells showed a low toxicity and a suitable biocompatibility. The MTT assay of the Que-loaded NPs possesses enhanced antitumor activity over the free Que. Copyright © 2018 Elsevier B.V. All rights reserved.

Evaluating Activated Carbon Adsorption of Dissolved Organic Matter and Micropollutants Using Fluorescence Spectroscopy.

PubMed

Shimabuku, Kyle K; Kennedy, Anthony M; Mulhern, Riley E; Summers, R Scott

2017-03-07

Dissolved organic matter (DOM) negatively impacts granular activated carbon (GAC) adsorption of micropollutants and is a disinfection byproduct precursor. DOM from surface waters, wastewater effluent, and 1 kDa size fractions were adsorbed by GAC and characterized using fluorescence spectroscopy, UV-absorption, and size exclusion chromatography (SEC). Fluorescing DOM was preferentially adsorbed relative to UV-absorbing DOM. Humic-like fluorescence (peaks A and C) was selectively adsorbed relative to polyphenol-like fluorescence (peaks T and B) potentially due to size exclusion effects. In the surface waters and size fractions, peak C was preferentially removed relative to peak A, whereas the reverse was found in wastewater effluent, indicating that humic-like fluorescence is associated with different compounds depending on DOM source. Based on specific UV-absorption (SUVA), aromatic DOM was preferentially adsorbed. The fluorescence index (FI), if interpreted as an indicator of aromaticity, indicated the opposite but exhibited a strong relationship with average molecular weight, suggesting that FI might be a better indicator of DOM size than aromaticity. The influence of DOM intermolecular interactions on adsorption were minimal based on SEC analysis. Fluorescence parameters captured the impact of DOM size on the fouling of 2-methylisoborneol and warfarin adsorption and correlated with direct competition and pore blockage indicators.

Spatial and seasonal changes in optical properties of autochthonous and allochthonous chromophoric dissolved organic matter in a stratified mountain lake.

PubMed

Bracchini, Luca; Dattilo, Arduino Massimo; Hull, Vincent; Loiselle, Steven Arthur; Nannicini, Luciano; Picchi, Maria Pia; Ricci, Maso; Santinelli, Chiara; Seritti, Alfredo; Tognazzi, Antonio; Rossi, Claudio

2010-03-01

In this study, we present results on seasonal and spatial changes in CDOM absorption and fluorescence (fCDOM) in a deep mountain lake (Salto Lake, Italy). A novel approach was used to describe the shape of CDOM absorption between 250-700 nm (distribution of the spectral slope, S(lambda)) and a new fluorescence ratio is used to distinguish between humic and amino acid-like components. Solar ultraviolet irradiance, dissolved organic carbon (DOC), DOM fluorescence and absorption measurements were analysed and compared to other physicochemical parameters. We show that in the UV-exposed mixed layer: (i) fluorescence by autochthonous amino acid-like CDOM, (ii) values of S(lambda) across UV-C and UV-B wavebands increased during the summer months, whereas (i) average molar absorption coefficient and (ii) fluorescence by allochthonous humic CDOM decreased. In the unexposed deep layer of the water column (and in the entire water column in winter), humic-like CDOM presented high values of molar absorption coefficients and low values of S(lambda). UV attenuation coefficients correlated with both chlorophyll a concentrations and CDOM absorption. In agreement with changes in CDOM, minimal values in UV attenuation were found in summer. The S(lambda) curve was used as a signature of the mixture between photobleached and algal-derived CDOM with respect to the unexposed chromophoric dissolved compounds in this thermal stratified lake. Furthermore, S(lambda) curves were useful to distinguish between low and high molecular weight CDOM.

Photoswitchable non-fluorescent thermochromic dye-nanoparticle hybrid probes.

PubMed

Harrington, Walter N; Haji, Mwafaq R; Galanzha, Ekaterina I; Nedosekin, Dmitry A; Nima, Zeid A; Watanabe, Fumiya; Ghosh, Anindya; Biris, Alexandru S; Zharov, Vladimir P

2016-11-08

Photoswitchable fluorescent proteins with controllable light-dark states and spectral shifts in emission in response to light have led to breakthroughs in the study of cell biology. Nevertheless, conventional photoswitching is not applicable for weakly fluorescent proteins and requires UV light with low depth penetration in bio-tissue. Here we introduce a novel concept of photoswitchable hybrid probes consisting of thermochromic dye and absorbing nanoparticles, in which temperature-sensitive light-dark states and spectral shifts in absorption can be switched through controllable photothermal heating of doped nanoparticles. The proof-of-concept is demonstrated through the use of two different types of temperature-sensitive dyes doped with magnetic nanoparticles and reversibly photoswitched by a near-infrared laser. Photoacoustic imaging revealed the high contrast of these probes, which is sufficient for their visualization in cells and deep tissue. Our results suggest that these new photoswitchable multicolour probes can be used for multimodal cellular diagnostics and potentially for magnetic and photothermal therapy.

A highly selective long-wavelength fluorescent probe for hydrazine and its application in living cell imaging

NASA Astrophysics Data System (ADS)

Hao, Yuanqiang; Zhang, Yintang; Ruan, Kehong; Meng, Fanteng; Li, Ting; Guan, Jinsheng; Du, Lulu; Qu, Peng; Xu, Maotian

2017-09-01

A highly selective long-wavelength turn-on fluorescent probe has been developed for the detection of N2H4. The probe was prepared by conjugation the tricyanofuran-based D-π-A system with a recognizing moiety of acetyl group. In the presence of N2H4, the probe can be effectively hydrazinolysized and produce a turn-on fluorescent emission at 610 nm as well as a large red-shift in the absorption spectrum corresponding to a color change from yellow to blue. The sensing mechanism was confirmed by HPLC, MS, UV-vis, emission spectroscopic and theoretical calculation studies. The probe displayed high selectivity and sensitivity for N2H4 with a LOD (limit of detection) of 0.16 μM. Moreover, the probe was successfully utilized for the detection of hydrazine in living cells.

Concentration, size, and excitation power effects on fluorescence from microdroplets and microparticles containing tryptophan and bacteria

NASA Astrophysics Data System (ADS)

Fell, Nicholas F., Jr.; Pinnick, Ronald G.; Hill, Steven C.; Videen, Gorden W.; Niles, Stanley; Chang, Richard K.; Holler, Stephen; Pan, Yongle; Bottiger, Jerold R.; Bronk, Burt V.

1999-01-01

Our group has been developing a system for single-particle fluorescence detection of aerosolized agents. This paper describes the most recent steps in the evolution of this system. The effects of fluorophore concentrations, droplet size, and excitation power have also been investigated with microdroplets containing tryptophan in water to determine the effects of these parameters on our previous results. The vibrating orifice droplet generator was chosen for this study base don its ability to generate particles of well- known and reproducible size. The power levels required to reach saturation and photodegradation were determined. In addition, the collection of fluorescence emission was optimized through the use of a UV achromatic photographic lens. This arrangement permitted collection of images of the droplet stream. Finally, the use of a dual-beam, conditional firing scheme facilitated the collection of improved signal- to-noise single-shot spectra from individual biological particles.

Photoswitchable non-fluorescent thermochromic dye-nanoparticle hybrid probes

NASA Astrophysics Data System (ADS)

Harrington, Walter N.; Haji, Mwafaq R.; Galanzha, Ekaterina I.; Nedosekin, Dmitry A.; Nima, Zeid A.; Watanabe, Fumiya; Ghosh, Anindya; Biris, Alexandru S.; Zharov, Vladimir P.

2016-11-01

Photoswitchable fluorescent proteins with controllable light-dark states and spectral shifts in emission in response to light have led to breakthroughs in the study of cell biology. Nevertheless, conventional photoswitching is not applicable for weakly fluorescent proteins and requires UV light with low depth penetration in bio-tissue. Here we introduce a novel concept of photoswitchable hybrid probes consisting of thermochromic dye and absorbing nanoparticles, in which temperature-sensitive light-dark states and spectral shifts in absorption can be switched through controllable photothermal heating of doped nanoparticles. The proof-of-concept is demonstrated through the use of two different types of temperature-sensitive dyes doped with magnetic nanoparticles and reversibly photoswitched by a near-infrared laser. Photoacoustic imaging revealed the high contrast of these probes, which is sufficient for their visualization in cells and deep tissue. Our results suggest that these new photoswitchable multicolour probes can be used for multimodal cellular diagnostics and potentially for magnetic and photothermal therapy.

The onset of anthracene phototoxicity to Lemna gibba and the protective effects of humic acid

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gensemer, R.W.; Dixon, D.G.; Greenberg, B.M.

1994-12-31

The toxicity of anthracene to the freshwater duckweed Lemna gibba is strongly photo-induced in the presence of light containing natural levels of ultraviolet (UV) radiation. This was demonstrated using 8-day static renewal bioassays at an anthracene concentration of 2 mg-L{sup {minus}1}. Plants were incubated under simulated solar radiation (SSR) which mimics UV levels found in natural sunlight at a visible:UV-A:UV-B ratio of 100:10:1. Anthracene phototoxicity was expressed as inhibition of population growth and fluorescence induction decreases in chlorophyll content, and changes in low-temperature chlorophyll fluorescence emission scans. Furthermore, adding 6.2 mg-L-1 of an artificial humic acid ameliorated anthracene phototoxicity evenmore » though HA is also photo modified by UV light. However, anthracene inhibited photosynthesis days before the endpoint assays were performed. Therefore, the authors repeated these experiments at short time intervals following exposure to both light and chemical. Anthracene phototoxicity occurred after only 1 hour as detected by chlorophyll fluorescence induction, whereas chlorophyll contents and low-temperature fluorescence emission scans were not affected until 24--48 hours, respectively. Humic acid again ameliorated anthracene toxicity by delaying the negative physiological events by approximately 24 hours.« less

The injury and cumulative effects on human skin by UV exposure from artificial fluorescence emission.

PubMed

Tian, Yan; Liu, Wei; Niu, TianHui; Dai, CaiHong; Li, Xiaoxin; Cui, Caijuan; Zhao, Xinyan; E, Yaping; Lu, Hui

2014-01-01

The injury and cumulative effects of UV emission from fluorescence lamp were studied. UV intensity from fluorescence lamp was measured, and human skin samples (hips, 10 volunteers) were exposed to low-dose UV irradiation (three times per week for 13 consecutive weeks). Three groups were examined: control group without UV radiation; low-dose group with a cumulative dose of 50 J cm(-2) which was equivalent to irradiation of the face during indoor work for 1.5 years; and high-dose group with 1000 J cm(-2) cumulative dose equivalent to irradiation of the face during outdoor activities for 1 year. Specific indicators were measured before and after UVA irradiation. The findings showed that extending the low-dose UVA exposure decreased the skin moisture content and increased the transepidermal water loss as well as induced skin color changes (decreased L* value, increased M index). Furthermore, irradiated skin showed an increased thickness of cuticle and epidermis, skin edema, light color and unclear staining collagen fibers in the dermis, and elastic fiber fragmentation. In addition, MMP-1, p53 and SIRT1 expression was also increased. Long-term exposure of low-dose UVA radiation enhanced skin photoaging. The safety of the fluorescent lamp needs our attention. © 2014 The American Society of Photobiology.

Real-time measurement of UV-inactivated Escherichia coli bacterial particles by electrospray-assisted UVAPS spectrometry.

PubMed

Jung, Jae Hee; Lee, Jung Eun; Bae, Gwi Nam

2011-08-01

The ultraviolet aerodynamic particle sizer (UVAPS) is a novel commercially available aerosol spectrometer for real-time continuous monitoring of viable bioaerosols, based on fluorescence from living microorganisms. In a previous study, we developed an electrospray-assisted UVAPS using biological electrospray techniques, which have the advantage of generating non-agglomerated single particles by the repulsive electrical forces. With this electrospraying of suspensions containing microorganisms, the analytical system can supply more accurate and quantitative information about living microorganisms than with conventional aerosolization. Using electrospray-assisted UVAPS, we investigated the characteristics of bacterial particles with various viabilities in real-time. Escherichia coli was used as the test microorganism, and its initial viability was controlled by the degree of exposure to UV irradiation. In the stable cone-jet domain, the particle size distributions of test bacterial particles remained almost uniform regardless of the degree of UV inactivation. However, the fluorescence spectra of the bacterial particles changed with the degree of UV inactivation. The fluorescence characteristics of UV-inactivated bacterial particles tended to show a similar decline with viability, determined by the sampling and culture method, although the percentage showing fluorescence was higher than that showing viability. Copyright © 2011 Elsevier B.V. All rights reserved.

Immuno-magnetic beads-based extraction-capillary zone electrophoresis-deep UV laser-induced fluorescence analysis of erythropoietin.

PubMed

Wang, Heye; Dou, Peng; Lü, Chenchen; Liu, Zhen

2012-07-13

Erythropoietin (EPO) is an important glycoprotein hormone. Recombinant human EPO (rhEPO) is an important therapeutic drug and can be also used as doping reagent in sports. The analysis of EPO glycoforms in pharmaceutical and sports areas greatly challenges analytical scientists from several aspects, among which sensitive detection and effective and facile sample preparation are two essential issues. Herein, we investigated new possibilities for these two aspects. Deep UV laser-induced fluorescence detection (deep UV-LIF) was established to detect the intrinsic fluorescence of EPO while an immuno-magnetic beads-based extraction (IMBE) was developed to specifically extract EPO glycoforms. Combined with capillary zone electrophoresis (CZE), CZE-deep UV-LIF allows high resolution glycoform profiling with improved sensitivity. The detection sensitivity was improved by one order of magnitude as compared with UV absorbance detection. An additional advantage is that the original glycoform distribution can be completely preserved because no fluorescent labeling is needed. By combining IMBE with CZE-deep UV-LIF, the overall detection sensitivity was 1.5 × 10⁻⁸ mol/L, which was enhanced by two orders of magnitude relative to conventional CZE with UV absorbance detection. It is applicable to the analysis of pharmaceutical preparations of EPO, but the sensitivity is insufficient for the anti-doping analysis of EPO in blood and urine. IMBE can be straightforward and effective approach for sample preparation. However, antibodies with high specificity were the key for application to urine samples because some urinary proteins can severely interfere the immuno-extraction. Copyright © 2012 Elsevier B.V. All rights reserved.

Improved sensing using simultaneous deep-UV Raman and fluorescence detection-II

NASA Astrophysics Data System (ADS)

Hug, W. F.; Bhartia, R.; Sijapati, K.; Beegle, L. W.; Reid, R. D.

2014-05-01

Photon Systems in collaboration with JPL is continuing development of a new technology robot-mounted or hand-held sensor for reagentless, short-range, standoff detection and identification of trace levels chemical, biological, and explosive (CBE) materials on surfaces. This deep ultraviolet CBE sensor is the result of Army STTR and DTRA programs. The evolving 10 to 15 lb, 20 W, sensor can discriminate CBE from background clutter materials using a fusion of deep UV excited resonance Raman (RR) and laser induced native fluorescence (LINF) emissions collected is less than 1 ms. RR is a method that provides information about molecular bonds, while LINF spectroscopy is a much more sensitive method that provides information regarding the electronic configuration of target molecules. Standoff excitation of suspicious packages, vehicles, persons, and other objects that may contain hazardous materials is accomplished using excitation in the deep UV where there are four main advantages compared to near-UV, visible or near-IR counterparts. 1) Excited between 220 and 250 nm, Raman emission occur within a fluorescence-free region of the spectrum, eliminating obscuration of weak Raman signals by fluorescence from target or surrounding materials. 2) Because Raman and fluorescence occupy separate spectral regions, detection can be done simultaneously, providing an orthogonal set of information to improve both sensitivity and lower false alarm rates. 3) Rayleigh law and resonance effects increase Raman signal strength and sensitivity of detection. 4) Penetration depth into target in the deep UV is short, providing spatial/spectral separation of a target material from its background or substrate. 5) Detection in the deep UV eliminates ambient light background and enable daylight detection.

Optical spectroscopic methods for probing the conformational stability of immobilised enzymes.

PubMed

Ganesan, Ashok; Moore, Barry D; Kelly, Sharon M; Price, Nicholas C; Rolinski, Olaf J; Birch, David J S; Dunkin, Ian R; Halling, Peter J

2009-07-13

We report the development of biophysical techniques based on circular dichroism (CD), diffuse reflectance infrared Fourier transform (DRIFT) and tryptophan (Trp) fluorescence to investigate in situ the structure of enzymes immobilised on solid particles. Their applicability is demonstrated using subtilisin Carlsberg (SC) immobilised on silica gel and Candida antartica lipase B immobilised on Lewatit VP.OC 1600 (Novozyme 435). SC shows nearly identical secondary structure in solution and in the immobilised state as evident from far UV CD spectra and amide I vibration bands. Increased near UV CD intensity and reduced Trp fluorescence suggest a more rigid tertiary structure on the silica surface. After immobilised SC is inactivated, these techniques reveal: a) almost complete loss of near UV CD signal, suggesting loss of tertiary structure; b) a shift in the amide I vibrational band from 1658 cm(-1) to 1632 cm(-1), indicating a shift from alpha-helical structure to beta-sheet; c) a substantial blue shift and reduced dichroism in the far UV CD, supporting a shift to beta-sheet structure; d) strong increase in Trp fluorescence intensity, which reflects reduced intramolecular quenching with loss of tertiary structure; and e) major change in fluorescence lifetime distribution, confirming a substantial change in Trp environment. DRIFT measurements suggest that pressing KBr discs may perturb protein structure. With the enzyme on organic polymer it was possible to obtain near UV CD spectra free of interference by the carrier material. However, far UV CD, DRIFT and fluorescence measurements showed strong signals from the organic support. In conclusion, the spectroscopic methods described here provide structural information hitherto inaccessible, with their applicability limited by interference from, rather than the particulate nature of, the support material.

Photoabsorption and photodissociation of molecules important in the interstellar medium

NASA Technical Reports Server (NTRS)

Lee, Long C.; Suto, Masako

1991-01-01

The photoabsorption, photodissociation, and fluorescence cross sections of interstellar molecules are measured at 90 to 250 nm. These quantitative optical data are needed for the understanding of the formation and destruction processes of molecules under the intense interstellar UV radiation field. Research covering the following topics is presented: (1) fluorescences from photoexcitation of CH4, CH3OH, and CH3SH; (2) NO gamma emission from photoexcitation of NO; (3) photoexcitation cross sections of aromatic molecules; (4) IR emission from UV excitation of HONO2; (5) IR emission from UV excitation of benzene and methyl-derivitives; and (6) IR emission from UV excitation of polycyclic aromatic hydrocarbon molecules.

Measurements of Raman scattering in the middle ultraviolet band from persistent chemical warfare agents

NASA Astrophysics Data System (ADS)

Kullander, Fredrik; Landström, Lars; Lundén, Hampus; Mohammed, Abdesalam; Olofsson, Göran; Wästerby, Pär.

2014-05-01

The very low Raman scattering cross section and the fluorescence background limit the measuring range of Raman based instruments operating in the visible or infrared band. We are exploring if laser excitation in the middle ultraviolet (UV) band between 200 and 300 nm is useful and advantageous for detection of persistent chemical warfare agents (CWA) on various kinds of surfaces. The UV Raman scattering from tabun, mustard gas, VX and relevant simulants in the form of liquid surface contaminations has been measured using a laboratory experimental setup with a short standoff distance around 1 meter. Droplets having a volume of 1 μl were irradiated with a tunable pulsed laser swept within the middle UV band. A general trend is that the signal strength moves through an optimum when the laser excitation wavelength is swept between 240 and 300 nm. The signal from tabun reaches a maximum around 265 nm, the signal from mustard gas around 275 nm. The Raman signal from VX is comparably weak. Raman imaging by the use of a narrow bandpass UV filter is also demonstrated.

Near-infrared fluorescence amplified organic nanoparticles with aggregation-induced emission characteristics for in vivo imaging

NASA Astrophysics Data System (ADS)

Geng, Junlong; Zhu, Zhenshu; Qin, Wei; Ma, Lin; Hu, Yong; Gurzadyan, Gagik G.; Tang, Ben Zhong; Liu, Bin

2013-12-01

Near-infrared (NIR) fluorescence signals are highly desirable to achieve high resolution in biological imaging. To obtain NIR emission with high brightness, fluorescent nanoparticles (NPs) are synthesized by co-encapsulation of 2,3-bis(4-(phenyl(4-(1,2,2-triphenylvinyl)phenylamino)phenyl)fumaronitrile (TPETPAFN), a luminogen with aggregation-induced emission (AIE) characteristics, and a NIR fluorogen of silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) (NIR775) using 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] as the encapsulation matrix. The good spectral overlap between the emission of TPETPAFN and the absorption of NIR775 leads to efficient energy transfer, resulting in a 47-fold enhancement of the NIR775 emission intensity upon excitation of TPETPAFN at 510 nm as compared to that upon direct excitation of NIR775 at 760 nm. The obtained fluorescent NPs show sharp NIR emission with a band width of 20 nm, a large Stokes shift of 275 nm, good photostability and low cytotoxicity. In vivo imaging study reveals that the synthesized NPs are able to provide high fluorescence contrast in live animals. The Förster resonance energy transfer strategy overcomes the intrinsic limitation of broad emission spectra for AIE NPs, which opens new opportunities to synthesize organic NPs with high brightness and narrow emission for potential applications in multiplex sensing and imaging.Near-infrared (NIR) fluorescence signals are highly desirable to achieve high resolution in biological imaging. To obtain NIR emission with high brightness, fluorescent nanoparticles (NPs) are synthesized by co-encapsulation of 2,3-bis(4-(phenyl(4-(1,2,2-triphenylvinyl)phenylamino)phenyl)fumaronitrile (TPETPAFN), a luminogen with aggregation-induced emission (AIE) characteristics, and a NIR fluorogen of silicon 2,3-naphthalocyanine bis(trihexylsilyloxide) (NIR775) using 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] as the encapsulation matrix. The good spectral overlap between the emission of TPETPAFN and the absorption of NIR775 leads to efficient energy transfer, resulting in a 47-fold enhancement of the NIR775 emission intensity upon excitation of TPETPAFN at 510 nm as compared to that upon direct excitation of NIR775 at 760 nm. The obtained fluorescent NPs show sharp NIR emission with a band width of 20 nm, a large Stokes shift of 275 nm, good photostability and low cytotoxicity. In vivo imaging study reveals that the synthesized NPs are able to provide high fluorescence contrast in live animals. The Förster resonance energy transfer strategy overcomes the intrinsic limitation of broad emission spectra for AIE NPs, which opens new opportunities to synthesize organic NPs with high brightness and narrow emission for potential applications in multiplex sensing and imaging. Electronic supplementary information (ESI) available: Characterization of AIE properties of TPETPAFN, UV-vis spectra of NPs, PL spectra comparison upon excitation at the donor and receptor absorbance maxima, ex vivo fluorescence imaging of mice organs. See DOI: 10.1039/c3nr04243j

MnFe2O4/CdSe magneto-fluorescent nanocomposite for possible biomedical applications

NASA Astrophysics Data System (ADS)

Chandunika, R. K.; Vijayaraghavan, R.; Sahu, Niroj Kumar

2018-04-01

Acombined superparamagnetic and fluorescent MnFe2O4/CdSe multifunctional nanocompositehas been prepared by suitable surface functionalizationswith citric acid, polyethyleneimine(PEI) and thioglycolic acid (ThA).and the samples were characterized by XRD, FT-IR, TEM, Zeta Potential, VSM, UV-Vis and PL spectroscopy. MnFe2O4 crystalizes with average size of 38.6 nm whereas CdSe with average size of 2.03 nm. In composite, the CdSe quantum dots (QD) are homogeneously distributed in the matrix of porous MnFe2O4 nanoparticles. Thenanocomposites are well dispersed in aqueous solvent and possess significant magnetic and luminescence properties which may be utilised for magnetic resonance imaging and luminescence tagging of biomolecules.

2D imaging and 3D sensing data acquisition and mutual registration for painting conservation

NASA Astrophysics Data System (ADS)

Fontana, Raffaella; Gambino, Maria Chiara; Greco, Marinella; Marras, Luciano; Pampaloni, Enrico M.; Pelagotti, Anna; Pezzati, Luca; Poggi, Pasquale

2004-12-01

We describe the application of 2D and 3D data acquisition and mutual registration to the conservation of paintings. RGB color image acquisition, IR and UV fluorescence imaging, together with the more recent hyperspectral imaging (32 bands) are among the most useful techniques in this field. They generally are meant to provide information on the painting materials, on the employed techniques and on the object state of conservation. However, only when the various images are perfectly registered on each other and on the 3D model, no ambiguity is possible and safe conclusions may be drawn. We present the integration of 2D and 3D measurements carried out on two different paintings: "Madonna of the Yarnwinder" by Leonardo da Vinci, and "Portrait of Lionello d'Este", by Pisanello, both painted in the XV century.

2D imaging and 3D sensing data acquisition and mutual registration for painting conservation

NASA Astrophysics Data System (ADS)

Fontana, Raffaella; Gambino, Maria Chiara; Greco, Marinella; Marras, Luciano; Pampaloni, Enrico M.; Pelagotti, Anna; Pezzati, Luca; Poggi, Pasquale

2005-01-01

We describe the application of 2D and 3D data acquisition and mutual registration to the conservation of paintings. RGB color image acquisition, IR and UV fluorescence imaging, together with the more recent hyperspectral imaging (32 bands) are among the most useful techniques in this field. They generally are meant to provide information on the painting materials, on the employed techniques and on the object state of conservation. However, only when the various images are perfectly registered on each other and on the 3D model, no ambiguity is possible and safe conclusions may be drawn. We present the integration of 2D and 3D measurements carried out on two different paintings: "Madonna of the Yarnwinder" by Leonardo da Vinci, and "Portrait of Lionello d'Este", by Pisanello, both painted in the XV century.

Transient Fluorescence Spectroscopy and laser induced fluorescence lifetimes of terbium doped dipicolinic acid

NASA Astrophysics Data System (ADS)

Makoui, Anali

We have investigated the use of deep UV laser induced fluorescence for the sensitive detection and spectroscopic lifetime studies of terbium doped dipicolinic acid (DPA-Tb) and used this to study the optical characteristics of DPA which is a chemical surrounding most bacterial spores. Background absorption spectra, fluorescence spectra, and Excitation Emission Matrix (EEM) spectra were made of the DPA-Tb complex, using both fixed 266 nm wavelength and tunable (220 nm--280 nm) UV laser excitations. Of importance, the fluorescence lifetimes of the four main fluorescence peaks (488 nm, 543 nm, 581 nm, and 618 nm) of the DPA-Tb complex have been measured for the first time to our knowledge. The lifetimes of all the fluorescing lines have been measured as a function of DPA-Tb concentration, solvent pH, and solvent composition, including that for the weakest fluorescing line of DPA-Tb at 618 nm. In addition, a new spectroscopic lifetime measurement technique, which we call "Transient Fluorescence Spectroscopy", was developed. In this technique, a weak, quasi-CW, amplitude modulated UV laser (8.5 kHz) was used to measure the lifetimes of the fluorescence lines, and yields insight into energy transfer and excitation lifetimes within the system. This technique is especially useful when a high power laser is not either available or not suitable. In the latter case, this would be when a high power pulsed deep-UV laser could produce bleaching or destruction of the biological specimen. In addition, this technique simulated the excitation and fluorescence emission of the DPA-Tb using a 4-level energy model, and solved the dynamic transient rate equations to predict the temporal behavior of the DPA-Tb emitted fluorescence. Excellent agreement between the experiments and the simulation were found. This technique has the potential to provide a more accurate value for the fluorescence lifetime values. In addition, with the use of asymmetric excitation waveforms, the dynamic transient rate equation analysis may allow for detailed studies of selected transfer mechanisms in a wide range of other spectroscopic applications including rare-earth solid-state lasing materials and biological samples.

Contributions of visible and ultraviolet parts of sunlight to photoinhibition.

PubMed

Hakala-Yatkin, Marja; Mäntysaari, Mika; Mattila, Heta; Tyystjärvi, Esa

2010-10-01

Photoinhibition is light-induced inactivation of PSII, and action spectrum measurements have shown that UV light causes photoinhibition much more efficiently than visible light. In the present study, we quantified the contribution of the UV part of sunlight in photoinhibition of PSII in leaves. Greenhouse-grown pumpkin leaves were pretreated with lincomycin to block the repair of photoinhibited PSII, and exposed to sunlight behind a UV-permeable or UV-blocking filter. Oxygen evolution and Chl fluorescence measurements showed that photoinhibition proceeds 35% more slowly under the UV-blocking than under the UV-permeable filter. Experiments with a filter that blocks UV-B but transmits UV-A and visible light revealed that UV-A light is almost fully responsible for the UV effect. The difference between leaves illuminated through a UV-blocking and UV-transparent filter disappeared when leaves of field-grown pumpkin plants were used. Thylakoids isolated from field-grown and greenhouse-grown plants were equally sensitive to UV light, and measurements of UV-induced fluorescence from leaves indicated that the protection of the field-grown plants was caused by substances that block the passage of UV light to the chloroplasts. Thus, the UV part of sunlight, especially the UV-A part, is potentially highly important in photoinhibition of PSII but the UV-screening compounds of plant leaves may offer almost complete protection against UV-induced photoinhibition.

Application of UV and Fluorescence Indices for Assessing the Performance of Ozonation Process: Towards Smart Water Treatment

NASA Astrophysics Data System (ADS)

Li, Wen-Tao; Abbt-Braun, Gudrun; Dodd, Michael; Korshin, Gregory; Li, Ai-Min

2017-04-01

The UV absorbance and fluorescence indices were comprehensively studied as surrogate indicators for assessing the degradation of dissolved organic matter (DOM), the formation of bromate and biodegradable dissolved organic carbon (BDOC) and the elimination of trace organic contaminants (TOrCs) during the ozonation of surface water and wastewater effluent. Spectroscopic monitoring was carried out using benchtop UV/Vis and fluorescence spectrophotometers and a newly developed miniature LED UV/fluorescence sensor capable of rapidly measuring UVA280 and protein-like and humic-like fluorescence. With the increase of O3/DOC mass ratio, the plots of BDOC formation were characterized three phases of initial lag, transition slope and final plateau. With the decrease of UV absorbance and fluorescence, BDOC concentrations initially increased slowly and then rose more noticeably. Inflection points in plots of BDOC versus changes of spectroscopic indicators were close to 35-45% loss of UVA254 or UVA280 and 75-85% loss of humic-like fluorescence. According to the data from size exclusion chromatography (SEC) with organic carbon detection and 2D synchronous correlation analyses, DOM fractions assigned to operationally defined large biopolymers (apparent molecular weight, AMW>20 kDa) and medium AMW humic substances (AMW 5.5-20 kDa) were transformed into medium-size building blocks (AMW 3-5.5 kDa) and other smaller AMW species (AMW<3 kDa) associated with BDOC at increasing O3/DOC ratios. Appreciable bromate formation was observed only after the values of UVA254, UVA280 and humic-like fluorescence in O3-treated samples were decreased by 45-55%, 50-60% and 86-92% relative to their respective initial levels. No significant differences in plots of bromate concentrations versus decreases of humic-like fluorescence were observed for surface water and wastewater effluent samples. This was in contrast with the plots of bromate concentration versus UVA254 and UVA280 which exhibited sensitivity to varying initial bromide concentrations in the investigated water matrixes. For TOrCs, their removal rates were well correlated with the decrease of the LED UV/fluorescence signals, and their elimination patterns were mainly determined by their reactivity with O3 and hydroxyl radicals. At approximately 50 % reduction of humic-like fluorescence almost complete oxidation of TOrCs of group I (e.g. carbamazepine) and II (e.g. gemfibrozil) was reached, a similar removal percentage (25-75 %) of TOrCs of group III (e.g. DEET) and IV (e.g. atrazine), and a poor removal percentage (< 25%) of group V (e.g. TCPP). In another way, 90% reduction of humic-like fluorescence could reach the sufficient elimination of most TOrCs. These results suggest that measurements of humic-like fluorescence can provide a useful supplement to UVA indices for characterization of ozonation processes.

Degradation of microbial fluorescence biosignatures by solar ultraviolet radiation on Mars

NASA Astrophysics Data System (ADS)

Dartnell, Lewis R.; Patel, Manish R.

2014-04-01

Recent and proposed robotic missions to Mars are equipped with implements to expose or excavate fresh material from beneath the immediate surface. Once brought into the open, any organic molecules or potential biosignatures of present or past life will be exposed to the unfiltered solar ultraviolet (UV) radiation and face photolytic degradation over short time courses. The key question, then, is what is the window of opportunity for detection of recently exposed samples during robotic operations? Detection of autofluorescence has been proposed as a simple method for surveying or triaging samples for organic molecules. Using a Mars simulation chamber we conduct UV exposures on thin frozen layers of two model microorganisms, the radiation-resistant polyextremophile Deinococcus radiodurans and the cyanobacterium Synechocystis sp. PCC 6803. Excitation-emission matrices (EEMs) are generated of the full fluorescence response to quantify the change in signal of different cellular fluorophores over Martian equivalent time. Fluorescence of Deinococcus cells, protected by a high concentration of carotenoid pigments, was found to be relatively stable over 32 h of Martian UV irradiation, with around 90% of the initial signal remaining. By comparison, fluorescence from protein-bound tryptophan in Synechocystis is much more sensitive to UV photodegradation, declining to 50% after 64 h exposure. The signal most readily degraded by UV irradiation is fluorescence of the photosynthetic pigments - diminished to only 35% after 64 h. This sensitivity may be expected as the biological function of chlorophyll and phycocyanin is to optimize the harvesting of light energy and so they are readily photobleached. A significant increase in a ~450 nm emission feature is interpreted as accumulation of fluorescent cellular degradation products from photolysis. Accounting for diurnal variation in Martian sunlight, this study calculates that frozen cellular biosignatures would remain detectable by fluorescence for at least several sols; offering a sufficient window for robotic exploration operations.

Spectral enhancement of leucocrystal violet treated footwear impression evidence in blood.

PubMed

Spence, Lindsay; Asmussen, Gary

2003-03-27

The results presented demonstrate the capacity for spectral enhancement to substantially improve the forensic examination of footwear impressions in blood treated with leucocrystal violet (LCV). The UV-Vis absorption spectra were generated of (i) an aqueous solution of leucocrystal violet, (ii) leucocrystal violet in 3% H(2)O(2), (iii) LCV working solution and (iv) whole blood added to LCV working solution. The resultant fluorescence emission spectra were subsequently generated (lambda(ex)=630nm, lambda(em)=661-900nm). The results indicate that the UV-Vis absorption spectra of an unbuffered solution of whole blood with LCV working solution produces a strong absorbance curve with a maxima at 630nm. Subsequent excitation at this wavelength and generation of the emission spectrum in the fluorescence mode indicates that a solution of whole blood added to LCV working solution is an extremely weak fluorophore. Therefore, to enable an adequate and timely enhancement of blood impression evidence treated with LCV utilising either visible fluorescence or infrared luminescence requires (i) selection of the most appropriate excitation wavelength (lambda(ex)) and emission wavelength (lambda(em)) with extremely narrow band pass filters, which in the absence of substrate matrix interference is excitation at 630nm producing the emission maxima at 665nm and (ii) a visual enhancement system such as a CCD colour IR video camera with image integration.

Calcium pump kinetics determined in single erythrocyte ghosts by microphotolysis and confocal imaging.

PubMed

Kubitscheck, U; Pratsch, L; Passow, H; Peters, R

1995-07-01

The activity of the plasma membrane calcium pump was measured in single cells. Human red blood cell ghosts were loaded with a fluorescent calcium indicator and either caged calcium and ATP (protocol A) or caged ATP and calcium (protocol B). In a suitably modified laser scanning microscope either calcium or ATP were released by a short UV light pulse. The time-dependent fluorescence intensity of the calcium indicator was then followed in single ghosts by repetitive confocal imaging. The fluorescence intensity was converted into calcium concentration, which in turn was used to derive the kinetic parameters of the calcium pump, the Michaelis-Menten constant Km, and the maximal transport rate vmax. Km and vmax values derived in this manner were 24 +/- 14 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol A, and 4 +/- 3 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol B, respectively. The difference between A and B is presumably caused by calmodulin, which is inactive in the experiments with protocol A. The possibilities to extend the new method to living nucleus-containing cells transiently transfected with mutants of the plasma membrane calcium pump are discussed.

7 CFR 93.11 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... fluoresce when viewed under UV light as follows: aflatoxin B1 and derivatives with a blue fluorescence, aflatoxin B2 with a blue-violet fluorescence, aflatoxin G1 with a green fluorescence, aflatoxin G2 with a green-blue fluorescence, aflatoxin M1 with a blue-violet fluorescence, and aflatoxin M2 with a violet...

7 CFR 93.11 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... fluoresce when viewed under UV light as follows: aflatoxin B1 and derivatives with a blue fluorescence, aflatoxin B2 with a blue-violet fluorescence, aflatoxin G1 with a green fluorescence, aflatoxin G2 with a green-blue fluorescence, aflatoxin M1 with a blue-violet fluorescence, and aflatoxin M2 with a violet...

7 CFR 93.11 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... fluoresce when viewed under UV light as follows: aflatoxin B1 and derivatives with a blue fluorescence, aflatoxin B2 with a blue-violet fluorescence, aflatoxin G1 with a green fluorescence, aflatoxin G2 with a green-blue fluorescence, aflatoxin M1 with a blue-violet fluorescence, and aflatoxin M2 with a violet...

7 CFR 93.11 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... fluoresce when viewed under UV light as follows: aflatoxin B1 and derivatives with a blue fluorescence, aflatoxin B2 with a blue-violet fluorescence, aflatoxin G1 with a green fluorescence, aflatoxin G2 with a green-blue fluorescence, aflatoxin M1 with a blue-violet fluorescence, and aflatoxin M2 with a violet...

7 CFR 93.11 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... fluoresce when viewed under UV light as follows: aflatoxin B1 and derivatives with a blue fluorescence, aflatoxin B2 with a blue-violet fluorescence, aflatoxin G1 with a green fluorescence, aflatoxin G2 with a green-blue fluorescence, aflatoxin M1 with a blue-violet fluorescence, and aflatoxin M2 with a violet...

Binding of caffeine with caffeic acid and chlorogenic acid using fluorescence quenching, UV/vis and FTIR spectroscopic techniques.

PubMed

Belay, Abebe; Kim, Hyung Kook; Hwang, Yoon-Hwae

2016-03-01

The interactions of caffeine (CF) with chlorogenic acid (CGA) and caffeic acid (CFA) were investigated by fluorescence quenching, UV/vis and Fourier transform infrared (FTIR) spectroscopic techniques. The results of the study indicated that the fluorescence quenching between caffeine and hydroxycinnamic acids could be rationalized in terms of static quenching or the formation of non-fluorescent CF-CFA and CF-CGA complexes. From fluorescence quenching spectral analysis, the quenching constant (KSV), quenching rate constant (kq), number of binding sites (n), thermodynamic properties and conformational changes of the interaction were determined. The quenching constants (KSV) between CF and CGA, CFA are 1.84 × 10(4) and 1.04 × 10(4) L/mol at 298 K and their binding site n is ~ 1. Thermodynamic parameters determined using the Van't Hoff equation indicated that hydrogen bonds and van der Waal's forces have a major role in the reaction of caffeine with caffeic acid and chlorogenic acid. The 3D fluorescence, UV/vis and FTIR spectra also showed that the binding of CF with CFA and CGA induces conformational changes in CFA and CGA. Copyright © 2015 John Wiley & Sons, Ltd.

[The research of UV-responsive sensitivity enhancement of fluorescent coating films by MgF2 layer].

PubMed

Lu, Zhong-Rong; Ni, Zheng-Ji; Tao, Chun-Xian; Hong, Rui-Jin; Zhang, Da-Wei; Huang, Yuan-Shen

2014-03-01

A low cost and less complicated expansion approach of wavelength responses with a Lumogen phosphor coating was adopted, as they increased the quantum efficiency of CCD and CMOS detectors in ultra-violet by absorbing UV light and then re emitting visible light. In this paper, the sensitivity enhancement of fluorescence coatings was studied by adding an anti-reflection film or barrier film to reduce the loss of the scattering and reflection on the incident interface. The Lumogen and MgF2/Lumogen film were deposited on quartz glasses by physical vacuum deposition. The surface morphology, transmittance spectrum, reflectance spectrum and fluorescence emission spectrum were obtained by atomic force microscope (AFM), spectrophotometer and fluorescence spectrometer, respectively. The results indicated that MgF2 film had obvious positive effect on reducing scattering and reflection loss in 500-700 nm, and enhancing the absorption of Lumogen coating in ultraviolet spectrum. Meanwhile, the fluorescent emission intensity had a substantial increase by smoothing the film surface and thus reducing the light scattering. At the same time, the MgF2 layer could protect Lumogen coating from damaging and contamination, which give a prolong lifetime of the UV-responsive CCD sensors with fluorescent coatings.

A Programmable DNA Double-Write Material: Synergy of Photolithography and Self-Assembly Nanofabrication.

PubMed

Song, Youngjun; Takahashi, Tsukasa; Kim, Sejung; Heaney, Yvonne C; Warner, John; Chen, Shaochen; Heller, Michael J

2017-01-11

We demonstrate a DNA double-write process that uses UV to pattern a uniquely designed DNA write material, which produces two distinct binding identities for hybridizing two different complementary DNA sequences. The process requires no modification to the DNA by chemical reagents and allows programmed DNA self-assembly and further UV patterning in the UV exposed and nonexposed areas. Multilayered DNA patterning with hybridization of fluorescently labeled complementary DNA sequences, biotin probe/fluorescent streptavidin complexes, and DNA patterns with 500 nm line widths were all demonstrated.

An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein

PubMed Central

Ando, Ryoko; Hama, Hiroshi; Yamamoto-Hino, Miki; Mizuno, Hideaki; Miyawaki, Atsushi

2002-01-01

We have cloned a gene encoding a fluorescent protein from a stony coral, Trachyphyllia geoffroyi, which emits green, yellow, and red light. The protein, named Kaede, includes a tripeptide, His-Tyr-Gly, that acts as a green chromophore that can be converted to red. The red fluorescence is comparable in intensity to the green and is stable under usual aerobic conditions. We found that the green-red conversion is highly sensitive to irradiation with UV or violet light (350–400 nm), which excites the protonated form of the chromophore. The excitation lights used to elicit red and green fluorescence do not induce photoconversion. Under a conventional epifluorescence microscope, Kaede protein expressed in HeLa cells turned red in a graded fashion in response to UV illumination; maximal illumination resulted in a 2,000-fold increase in the ratio of red-to-green signal. These color-changing properties provide a simple and powerful technique for regional optical marking. A focused UV pulse creates an instantaneous plane source of red Kaede within the cytosol. The red spot spreads rapidly throughout the cytosol, indicating its free diffusibility in the compartment. The extensive diffusion allows us to delineate a single neuron in a dense culture, where processes originating from many different somata are present. Illumination of a focused UV pulse onto the soma of a Kaede-expressing neuron resulted in filling of all processes with red fluorescence, allowing visualization of contact sites between the red and green neurons of interest. PMID:12271129

Allochthonous Addition of Meteoritic Organics to the Lunar Regolith

NASA Technical Reports Server (NTRS)

Thomas-Keprta, K. L.; Clemett, S.; Ross, D. K.; Le, L.; Rahman, Z.; McKay, D. S.; Gibson, E. K.; Gonzalez, C.

2013-01-01

Preparation of lunar samples 74220,861 was discussed in detail in [3, 4]. Our analysis sequence was as follows: optical microscopy, UV fluorescence imaging, -Raman, FESEM-EDX imaging and mapping, FETEMEDX imaging and mapping of a Focused Ion Beam (FIB) extracted section, and NanoSIMs analysis. We observed fluffytextured C-rich regions of interest (ROI) on three different volcanic glass beads. Each ROI was several m2 in size and fluoresced when exposed to UV. Using FESEM/EDX, the largest ROI measured 36 m and was located on an edge of a plateau located on the uppermost surface of the bead. The ROI was covered on one edge by a siliceous filament emanating from the plateau surface indicating it was attached to the bead while on the Moon. EDX mapping of the ROI shows it is composed primarily of heterogeneously distributed C. Embedded with the carbonaceous phase are localized concentrations of Si, Fe, Al and Ti indicating the presence of glass and/or minerals grains. -Raman showed strong D- and G-bands and their associated second order bands; intensity and location of these bands indicates the carbonaceous matter is structurally disorganized. A TEM thin section was extracted from the surface of a glass bead using FIB microscopy. High resolution TEM imaging and selected area electron diffraction demonstrate the carbonaceous layer to be amorphous; it lacked any long or short range order characteristic of micro- or nanocrystalline graphite. Additionally TEM imaging also revealed the presence of submicron mineral grains, typically < 50 nm in size, dispersed within the carbonaceous layer. NanoSIMs data will be presented and discussed at the meeting. Given the noted similarities between the carbonaceous matter present on 74220 glass beads and meteoritic kerogen, we suggest the allochthonous addition of meteoritic organics as the most probable source for the C-rich ROIs.

Ablation plume structure and dynamics in ambient gas observed by laser-induced fluorescence imaging spectroscopy

NASA Astrophysics Data System (ADS)

Miyabe, M.; Oba, M.; Iimura, H.; Akaoka, K.; Khumaeni, A.; Kato, M.; Wakaida, I.

2015-08-01

The dynamic behavior of an ablation plume in ambient gas has been investigated by laser-induced fluorescence imaging spectroscopy. The second harmonic beam from an Nd:YAG laser (0.5-6 J/cm2) was focused on a sintered oxide pellet or a metal chip of gadolinium. The produced plume was subsequently intersected with a sheet-shaped UV beam from a dye laser so that time-resolved fluorescence images were acquired with an intensified CCD camera at various delay times. The obtained cross-sectional images of the plume indicate that the ablated ground state atoms and ions of gadolinium accumulate in a hemispherical contact layer between the plume and the ambient gas, and a cavity containing a smaller density of ablated species is formed near the center of the plume. At earlier expansion stage, another luminous component also expands in the cavity so that it coalesces into the hemispherical layer. The splitting and coalescence for atomic plume occur later than those for ionic plume. Furthermore, the hemispherical layer of neutral atoms appears later than that of ions; however, the locations of the layers are nearly identical. This coincidence of the appearance locations of the layers strongly suggests that the neutral atoms in the hemispherical layer are produced as a consequence of three-body recombination of ions through collisions with gas atoms. The obtained knowledge regarding plume expansion dynamics and detailed plume structure is useful for optimizing the experimental conditions for ablation-based spectroscopic analysis.

Use of a Reflective Ultraviolet Imaging System (RUVIS) on Two-Dimensional Dust Impressions Created with Footwear on Multiple Substrates

NASA Astrophysics Data System (ADS)

Engelson, Brian Aaron

Footwear impression evidence in dust is often difficult to locate in ambient light and is a fragile medium that both collection and enhancement techniques can destroy or distort. The collection of footwear impression evidence always begins with non-destructive photographic techniques; however, current methods are limited to oblique lighting of the impression followed by an attempt to photograph in situ. For the vast majority of footwear impressions, an interactive collection method, and thus a potentially destructive procedure, is subsequently carried out to gather the evidence. Therefore, alternative non-destructive means for the preservation and enhancement of footwear impressions in dust merits further attention. Previous research performed with reflected ultraviolet (UV) photography and reflected ultraviolet imaging systems (RUVIS) has shown that there are additional non-destructive methodologies that can be applied to the search for and documentation of footwear impressions in dust. Unfortunately, these prior studies did not include robust comparisons to traditional oblique white light, instead choosing to focus on different UV wavelengths. This study, however, seeks to evaluate the use of a RUVIS device paired with a 254 nanometer (nm) UV light source to locate 2-D footwear impressions in dust on multiple substrates against standard oblique white light techniques and assess the visibility of the impression and amount of background interference present. The optimal angle of incident UV light for each substrate was also investigated. Finally, this study applied an image enhancement technique in order to evaluate its usefulness when looking at the visibility of a footwear impression and the amount of background interference present for enhanced white light and RUVIS pictures of footwear impressions in dust. A collection of eight different substrate types was gathered for investigation, including vinyl composition tile (VCT), ceramic tile, marble tile, magazine paper, steel sheet metal, vinyl flooring, wood flooring, and carpet. Heel impressions were applied to the various substrates utilizing vacuum collected dust and normal walking pressure. Each substrate was then explored and photographed in ambient fluorescent light, oblique white light at 0°, 15°, 30°, and 0° with the light source below the surface plane of the substrate, and 254 nm UV light at 0°, 15°, 30°, 45°, 60°, 75°, 90° and 0° with the light source below the surface plane of the substrate. All pictures were evaluated for clarity and visible detail of the footwear impression and the amount of background interference present, selecting for the best images within a lighting condition group. Additional intra- and intergroup comparisons were carried out to explore differences created by the various lighting conditions. Enhanced images were then created with the best scored pictures and evaluated for additional modifications in impression visibility and background interference. Photographs of footwear impressions in dust illuminated with ambient fluorescent light proved to be the most difficult conditions under which a footwear impression could be visualized. However, both oblique white light and 254 nm UV light lighting conditions showed improvements in either visualization or background dropout, or both, over ambient light conditions. An assessment of the white light and 254 nm UV light RUVIS images also demonstrated that the best angles for the light source for all substrates were oblique 0 and oblique 0° below the surface plane of the substrate lighting. It was found that white light photographs generally provided higher visibility ratings, while RUVIS 254 nm UV light photographs provided better grades for reducing background interference. Enhanced images of white light conditions provided generally poorer quality and quantity of details, while enhanced RUVIS images seemed to improve upon these areas. The use of a RUVIS to capture photographs of footwear impression evidence in dust was found to be a successful secondary non-destructive technique that can be paired with traditional oblique white light procedures. Additionally, the use of below the surface plane of the substrate lighting techniques were found to improve either visibility or background dropout, or both, over standard 0 oblique lighting, depending on the light source, and should be employed, when applicable. Finally, further investigation into digital photo-editing enhancement techniques for footwear impression evidence in dust is needed.

Prediction of BOD, COD, and total nitrogen concentrations in a typical urban river using a fluorescence excitation-emission matrix with PARAFAC and UV absorption indices.

PubMed

Hur, Jin; Cho, Jinwoo

2012-01-01

The development of a real-time monitoring tool for the estimation of water quality is essential for efficient management of river pollution in urban areas. The Gap River in Korea is a typical urban river, which is affected by the effluent of a wastewater treatment plant (WWTP) and various anthropogenic activities. In this study, fluorescence excitation-emission matrices (EEM) with parallel factor analysis (PARAFAC) and UV absorption values at 220 nm and 254 nm were applied to evaluate the estimation capabilities for biochemical oxygen demand (BOD), chemical oxygen demand (COD), and total nitrogen (TN) concentrations of the river samples. Three components were successfully identified by the PARAFAC modeling from the fluorescence EEM data, in which each fluorophore group represents microbial humic-like (C1), terrestrial humic-like organic substances (C2), and protein-like organic substances (C3), and UV absorption indices (UV(220) and UV(254)), and the score values of the three PARAFAC components were selected as the estimation parameters for the nitrogen and the organic pollution of the river samples. Among the selected indices, UV(220), C3 and C1 exhibited the highest correlation coefficients with BOD, COD, and TN concentrations, respectively. Multiple regression analysis using UV(220) and C3 demonstrated the enhancement of the prediction capability for TN.

Surface Nanobubbles Studied by Time-Resolved Fluorescence Microscopy Methods Combined with AFM: The Impact of Surface Treatment on Nanobubble Nucleation.

PubMed

Hain, Nicole; Wesner, Daniel; Druzhinin, Sergey I; Schönherr, Holger

2016-11-01

The impact of surface treatment and modification on surface nanobubble nucleation in water has been addressed by a new combination of fluorescence lifetime imaging microscopy (FLIM) and atomic force microscopy (AFM). In this study, rhodamine 6G (Rh6G)-labeled surface nanobubbles nucleated by the ethanol-water exchange were studied on differently cleaned borosilicate glass, silanized glass as well as self-assembled monolayers on transparent gold by combined AFM-FLIM. While the AFM data confirmed earlier reports on surface nanobubble nucleation, size, and apparent contact angles in dependence of the underlying substrate, the colocalization of these elevated features with highly fluorescent features observed in confocal intensity images added new information. By analyzing the characteristic contributions to the excited state lifetime of Rh6G in decay curves obtained from time-correlated single photon counting (TCSPC) experiments, the characteristic short-lived (<600 ps) component of could be associated with an emission at the gas-water interface. Its colocalization with nanobubble-like features in the AFM height images provides evidence for the observation of gas-filled surface nanobubbles. While piranha-cleaned glass supported nanobubbles, milder UV-ozone or oxygen plasma treatment afforded glass-water interfaces, where no nanobubbles were observed by combined AFM-FLIM. Finally, the number density of nanobubbles scaled inversely with increasing surface hydrophobicity.

Two-photon excitation in chip electrophoresis enabling label-free fluorescence detection in non-UV transparent full-body polymer chips.

PubMed

Geissler, David; Belder, Detlev

2015-12-01

One of the most commonly employed detection methods in microfluidic research is fluorescence detection, due to its ease of integration and excellent sensitivity. Many analytes though do not show luminescence when excited in the visible light spectrum, require suitable dyes. Deep-ultraviolet (UV) excitation (<300 nm) allows label-free detection of a broader range of analytes but also mandates the use of expensive fused silica glass, which is transparent to UV light. Herein, we report the first application of label-free deep UV fluorescence detection in non-UV transparent full-body polymer microfluidic devices. This was achieved by means of two-photon excitation in the visible range (λex = 532 nm). Issues associated with the low optical transmittance of plastics in the UV range were successfully circumvented in this way. The technique was investigated by application to microchip electrophoresis of small aromatic compounds. Various polymers, such as poly(methyl methacrylate), cyclic olefin polymer, and copolymer as well as poly(dimethylsiloxane) were investigated and compared with respect to achievable LOD and ruggedness against photodamage. To demonstrate the applicability of the technique, the method was also applied to the determination of serotonin and tryptamine in fruit samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ultraviolet-Visible and Fluorescence Spectroscopy Techniques Are Important Diagnostic Tools during the Progression of Atherosclerosis: Diet Zinc Supplementation Retarded or Delayed Atherosclerosis

PubMed Central

Abdelhalim, Mohamed Anwar K.; Moussa, Sherif A. Abdelmottaleb; AL-Mohy, Yanallah Hussain

2013-01-01

Background. In this study, we examined whether UV-visible and fluorescence spectroscopy techniques detect the progression of atherosclerosis in serum of rabbits fed on high-cholesterol diet (HCD) and HCD supplemented with zinc (HCD + Zn) compared with the control. Methods. The control rabbits group was fed on 100 g/day of normal diet. The HCD group was fed on Purina Certified Rabbit Chow supplemented with 1.0% cholesterol plus 1.0% olive oil (100 g/day) for the same period. The HCD + Zn group was fed on normal Purina Certified Rabbit Chow plus 1.0% cholesterol and 1.0% olive oil supplemented with 470 ppm Zn for the same feeding period. UV-visible and fluorescence spectroscopy and biochemistry in Rabbit's blood serum and blood hematology were measured in Rabbit's blood. Results. We found that the fluorescent peak of HCD shifted toward UV-visible wavelength compared with the control using fluorescent excitation of serum at 192 nm. In addition, they showed that supplementation of zinc (350 ppm) restored the fluorescent peak closely to the control. By using UV-visible spectroscopy approach, we found that the peak absorbance of HCD (about 280 nm) was higher than that of control and that zinc supplementation seemed to decrease the absorbance. Conclusions. This study demonstrates that ultraviolet-visible and fluorescence spectroscopy techniques can be applied as noninvasive techniques on a sample blood serum for diagnosing or detecting the progression of atherosclerosis. The Zn supplementation to rabbits fed on HCD delays or retards the progression of atherosclerosis. Inducing anemia in rabbits fed on HCD delays the progression of atherosclerosis. PMID:24350281

Interaction mechanisms between organic UV filters and bovine serum albumin as determined by comprehensive spectroscopy exploration and molecular docking.

PubMed

Ao, Junjie; Gao, Li; Yuan, Tao; Jiang, Gaofeng

2015-01-01

Organic UV filters are a group of emerging PPCP (pharmaceuticals and personal care products) contaminants. Current information is insufficient to understand the in vivo processes and health risks of organic UV filters in humans. The interaction mechanism of UV filters with serum albumin provides critical information for the health risk assessment of these active ingredients in sunscreen products. This study investigates the interaction mechanisms of five commonly used UV filters (2-hydroxy-4-methoxybenzophenone, BP-3; 2-ethylhexyl 4-methoxycinnamate, EHMC; 4-methylbenzylidene camphor, 4-MBC; methoxydibenzoylmethane, BDM; homosalate, HMS) with bovine serum albumin (BSA) by spectroscopic measurements of fluorescence, circular dichroism (CD), competitive binding experiments and molecular docking. Our results indicated that the fluorescence of BSA was quenched by these UV filters through a static quenching mechanism. The values of the binding constant (Ka) ranged from (0.78±0.02)×10(3) to (1.29±0.01)×10(5) L mol(-1). Further exploration by synchronous fluorescence and CD showed that the conformation of BSA was demonstrably changed in the presence of these organic UV filters. It was confirmed that the UV filters can disrupt the α-helical stability of BSA. Moreover, the results of molecular docking revealed that the UV filter molecule is located in site II (sub-domain IIIA) of BSA, which was further confirmed by the results of competitive binding experiments. In addition, binding occurred mainly through hydrogen bonding and hydrophobic interaction. This study raises critical concerns regarding the transportation, distribution and toxicity effects of organic UV filters in human body. Copyright © 2014 Elsevier Ltd. All rights reserved.

Monitoring exocytosis and release from individual mast cells by capillary electrophoresis and UV imaging microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yeung, E.S.; Lillard, S.J.; McCloskey, M.A.

1997-12-31

The complex temporal evolution of on-column exocytotic release of serotonin from individual peritoneal mast cells (RPMCs) was monitored by using capillary electrophoresis and UV imaging microscopy. Laser-induced native fluorescence detection with 275-nm excitation was used, and a detection limit of 1.7 amol (S/N = 3; rms) was obtained for serotonin. A physiological running buffer was used to ensure that the cell remained viable throughout. The secretagogue was polymyxin B sulfate (Pmx). Following the injection of a single mast cell into the capillary, electromigration of Pmx toward and past the cell induced degranulation and release of serotonin. The time course ofmore » release was registered in the electropherograms with subsecond resolution. Subsequent introduction of SDS caused the cell to lyse completely and allowed the residual serotonin to be quantified. The average amount of serotonin observed per RPMC was 1.6 {+-} 0.6 fmol; the average percentage of serotonin released was 28 {+-} 14%. Events that are consistent with released serontonin from single submicron granules (250 aL each) were evident, each of which contained an average amount of 5.9 {+-} 3 amol. Alternatively, UV movies can be taken of the entire event to provide temporal and spatial information.« less

Determining the Ice-binding Planes of Antifreeze Proteins by Fluorescence-based Ice Plane Affinity

PubMed Central

Basu, Koli; Garnham, Christopher P.; Nishimiya, Yoshiyuki; Tsuda, Sakae; Braslavsky, Ido; Davies, Peter

2014-01-01

Antifreeze proteins (AFPs) are expressed in a variety of cold-hardy organisms to prevent or slow internal ice growth. AFPs bind to specific planes of ice through their ice-binding surfaces. Fluorescence-based ice plane affinity (FIPA) analysis is a modified technique used to determine the ice planes to which the AFPs bind. FIPA is based on the original ice-etching method for determining AFP-bound ice-planes. It produces clearer images in a shortened experimental time. In FIPA analysis, AFPs are fluorescently labeled with a chimeric tag or a covalent dye then slowly incorporated into a macroscopic single ice crystal, which has been preformed into a hemisphere and oriented to determine the a- and c-axes. The AFP-bound ice hemisphere is imaged under UV light to visualize AFP-bound planes using filters to block out nonspecific light. Fluorescent labeling of the AFPs allows real-time monitoring of AFP adsorption into ice. The labels have been found not to influence the planes to which AFPs bind. FIPA analysis also introduces the option to bind more than one differently tagged AFP on the same single ice crystal to help differentiate their binding planes. These applications of FIPA are helping to advance our understanding of how AFPs bind to ice to halt its growth and why many AFP-producing organisms express multiple AFP isoforms. PMID:24457629

Determining the ice-binding planes of antifreeze proteins by fluorescence-based ice plane affinity.

PubMed

Basu, Koli; Garnham, Christopher P; Nishimiya, Yoshiyuki; Tsuda, Sakae; Braslavsky, Ido; Davies, Peter

2014-01-15

Antifreeze proteins (AFPs) are expressed in a variety of cold-hardy organisms to prevent or slow internal ice growth. AFPs bind to specific planes of ice through their ice-binding surfaces. Fluorescence-based ice plane affinity (FIPA) analysis is a modified technique used to determine the ice planes to which the AFPs bind. FIPA is based on the original ice-etching method for determining AFP-bound ice-planes. It produces clearer images in a shortened experimental time. In FIPA analysis, AFPs are fluorescently labeled with a chimeric tag or a covalent dye then slowly incorporated into a macroscopic single ice crystal, which has been preformed into a hemisphere and oriented to determine the a- and c-axes. The AFP-bound ice hemisphere is imaged under UV light to visualize AFP-bound planes using filters to block out nonspecific light. Fluorescent labeling of the AFPs allows real-time monitoring of AFP adsorption into ice. The labels have been found not to influence the planes to which AFPs bind. FIPA analysis also introduces the option to bind more than one differently tagged AFP on the same single ice crystal to help differentiate their binding planes. These applications of FIPA are helping to advance our understanding of how AFPs bind to ice to halt its growth and why many AFP-producing organisms express multiple AFP isoforms.

Neuroendocrine Tumor-Targeted Upconversion Nanoparticle-Based Micelles for Simultaneous NIR-Controlled Combination Chemotherapy and Photodynamic Therapy, and Fluorescence Imaging

PubMed Central

Chen, Guojun; Jaskula-Sztul, Renata; Esquibel, Corinne R.; Lou, Irene; Zheng, Qifeng; Dammalapati, Ajitha; Harrison, April; Eliceiri, Kevin W.; Tang, Weiping

2017-01-01

Although neuroendocrine tumors (NETs) are slow growing, they are frequently metastatic at the time of discovery and no longer amenable to curative surgery, emphasizing the need for the development of other treatments. In this study, multifunctional upconversion nanoparticle (UCNP)-based theranostic micelles are developed for NET-targeted and near-infrared (NIR)-controlled combination chemotherapy and photodynamic therapy (PDT), and bioimaging. The theranostic micelle is formed by individual UCNP functionalized with light-sensitive amphiphilic block copolymers poly(4,5-dimethoxy-2-nitrobenzyl methacrylate)-polyethylene glycol (PNBMA-PEG) and Rose Bengal (RB) photosensitizers. A hydrophobic anticancer drug, AB3, is loaded into the micelles. The NIR-activated UCNPs emit multiple luminescence bands, including UV, 540 nm, and 650 nm. The UV peaks overlap with the absorption peak of photocleavable hydrophobic PNBMA segments, triggering a rapid drug release due to the NIR-induced hydrophobic-to-hydrophilic transition of the micelle core and thus enabling NIR-controlled chemotherapy. RB molecules are activated via luminescence resonance energy transfer to generate 1O2 for NIR-induced PDT. Meanwhile, the 650 nm emission allows for efficient fluorescence imaging. KE108, a true pansomatostatin nonapeptide, as an NET-targeting ligand, drastically increases the tumoral uptake of the micelles. Intravenously injected AB3-loaded UCNP-based micelles conjugated with RB and KE108—enabling NET-targeted combination chemotherapy and PDT—induce the best antitumor efficacy. PMID:28989337

Visual Method for Detecting Contaminant on Dried Nutmeg Using Fluorescence Imaging

NASA Astrophysics Data System (ADS)

Dahlan, S. A.; Ahmad, U.; Subrata, I. D. M.

2018-05-01

Traditional practice of nutmeg sun-drying causes some fungi such as Aspergillus flavus to grow. One of the secondary metabolites of A. flavus named aflatoxin (AFs) is known to be carcinogenic, so the dried nutmeg kernel must be aflatoxin-free in the trading. Aflatoxin detection requires time and costly, make it difficult to conduct at the farmers level. This study aims to develop a simple and low-cost method to detect aflatoxin at the farmer level. Fresh nutmeg seeds were dried in two ways; sundried everyday (continuous), and sundried every two days (intermittent), both for around 18 days. The dried nutmeg seeds are then stored in a rice sack under normal conditions until the fungi grow, then they were opened and the images of kernels were captured using a CCD camera, with normal light and UV light sources. Visual observation on images captured in normal light source was able to detect the presence of fungi on dried kernels, by 28.0% for continuous and 26.2% for intermittent sun-drying. Visual observation on images captured in UV light source was able to detect the presence of aflatoxin on dried kernels, indicated by blue luminance on kernel, by 10.4% and 13.4% for continuous and intermittent sun-drying.

Combined "dual" absorption and fluorescence smartphone spectrometers.

PubMed

Arafat Hossain, Md; Canning, John; Ast, Sandra; Cook, Kevin; Rutledge, Peter J; Jamalipour, Abbas

2015-04-15

A combined "dual" absorption and fluorescence smartphone spectrometer is demonstrated. The optical sources used in the system are the white flash LED of the smartphone and an orthogonally positioned and interchangeable UV (λ_ex=370  nm) and blue (λ_ex=450  nm) LED. The dispersive element is a low-cost, nano-imprinted diffraction grating coated with Au. Detection over a 300 nm span with 0.42 nm/pixel resolution was carried out with the camera CMOS chip. By integrating the blue and UV excitation sources into the white LED circuitry, the entire system is self-contained within a 3D printed case and powered from the smartphone battery; the design can be scaled to add further excitation sources. Using a customized app, acquisition of absorption and fluorescence spectra are demonstrated using a blue-absorbing and green-emitting pH-sensitive amino-naphthalimide-based fluorescent probe and a UV-absorbing and blue-emitting Zn²⁺-sensitive fluoro-ionophore.

Impact of surface grafting density of PEG macromolecules on dually fluorescent silica nanoparticles used for the in vivo imaging of subcutaneous tumors.

PubMed

Adumeau, Laurent; Genevois, Coralie; Roudier, Lydia; Schatz, Christophe; Couillaud, Franck; Mornet, Stéphane

2017-06-01

In the context of systematically administered nanomedicines, the physicochemistry of NP surfaces must be controlled as a prerequisite to improve blood circulation time, and passive and active targeting. In particular, there is a real need to develop NP stealth and labelling for both in vivo and microscopic fluorescence imaging in a mice model. We have synthesized NIR/red dually fluorescent silica nanoparticles of 19nm covalently covered by a PEG layer of different grafting density in the brush conformational regime by using a reductive amination reaction. These particles were characterized by TEM, DRIFT, DLS, TGA, ζ potential measurements, UV-vis and fluorescence spectroscopy. Prostate tumors were generated in mice by subcutaneous injection of RM1-CMV-Fluc cells. Tumor growth was monitored by BLI after a D-luciferin injection. Four samples of PEGylated fluorescent NPs were individually intravenously injected into 6 mice (N=6, total 24 mice). Nanoparticle distribution was investigated using in vivo fluorescence reflectance imaging (FRI) over 48h and microscopy imaging was employed to localize the NPs within tumors in vitro. Fluorescent NP accumulation, due to the enhanced permeability and retention (EPR) effect, increases gradually as a function of increased PEG surface grafting density with a huge difference observed for the highest density grafting. For the highest grafting density, a blood circulation time of up to 24h was observed with a strong reduction in uptake by the liver. In vivo experimental results suggest that the biodistribution of NPs is very sensitive to slight variations in surface grafting density when the NPs present a high curvature radius. This study underlines the need to compensate a high curvature radius with a PEG-saturated NP surface to improve blood circulation and accumulation within tumors through the EPR effect. Dually fluorescent NPs PEGylated to saturation display physical properties useful for assessing the susceptibility of tumors to the EPR effect. Control of the physicochemical features of nanoparticle surfaces to improve blood circulation times and monitoring of the EPR effect. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader. Copyright © 2017 Elsevier B.V. All rights reserved.

Assessment of the Content of Fluorescent Tracer in Granular Feed Mixture.

PubMed

Matuszek, Dominika B; Wojtkiewicz, Krystian

2018-05-03

Background: This paper describes the use of fluorescence induced by UV radiation to evaluate the share of tracer in feed mixture. Methods: For the purpose of this study, three substances were used. They are as follows: Tinopal, Rhodamine B, and Uranine. Tracer in the form of maize or kardi was added to chicken feed before the mixing process. Grains used in the process were grinded in the mill sieve with a mesh size of 4 and 6 mm. The drawn samples of the mixture were illuminated with UV radiation to make grain tracer light, and then the photo was taken with a digital camera. The acquired images were analyzed with the use of a computer program running on the RGB color model, which was the way to obtain essential information about the percentage share of tracer. Results: It was observed that, in the case of kardi grains, the proposed method gives results significantly deviating from the verification method. Conclusions: Only the tests with the use of maize having an average particle diameter of 2.4 mm and tinted with the solution of Rhodamine B led to acceptable results (consensual with the predetermined verification level).

Demonstrating the Influence of UV Rays on Living Things.

ERIC Educational Resources Information Center

Morimoto, Kouichi

2002-01-01

Describes an experiment that introduces students to the different types of UV rays and their effects on living things by using appropriate teaching materials and equipment. Demonstrates the effects of exposure to UV-B (fluorescent) and UV-C (germicidal) lamps by using bananas, duckweed, and the fruit fly. (Contains 14 references.) (Author/YDS)

Application of portable online LED UV fluorescence sensor to predict the degradation of dissolved organic matter and trace organic contaminants during ozonation.

PubMed

Li, Wen-Tao; Majewsky, Marius; Abbt-Braun, Gudrun; Horn, Harald; Jin, Jing; Li, Qiang; Zhou, Qing; Li, Ai-Min

2016-09-15

This work aims to correlate signals of LED UV/fluorescence sensor with the degradation of dissolved organic matter (DOM) and trace-level organic contaminants (TOrCs) during ozonation process. Six sets of bench-scale ozonation kinetic experiments incorporated with three different water matrices and 14 TOrCs of different reactivity (group I ∼ V) were conducted. Calibrated by tryptophan and humic substances standards and verified by the lab benchtop spectroscopy, the newly developed portable/online LED sensor, which measures the UV280 absorbance, protein-like and humic-like fluorescence simultaneously, was feasible to monitor chromophores and fluorophores with good sensitivity and accuracy. The liquid chromatography with organic carbon detector combined with 2D synchronous correlation analysis further demonstrated how the DOM components of large molecular weight were transformed into small moieties as a function of the decrease of humic-like fluorescence. For TOrCs, their removal rates were well correlated with the decrease of the LED UV/fluorescence signals, and their elimination patterns were mainly determined by their reactivity with O3 and hydroxyl radicals. At approximately 50% reduction of humic-like fluorescence almost complete oxidation of TOrCs of group I and II was reached, a similar removal percentage (25-75%) of TOrCs of group III and IV, and a poor removal percentage (<25%) of group V. This study might contribute to the smart control of advanced oxidation processes for the water and wastewater treatment in the future. Copyright © 2016 Elsevier Ltd. All rights reserved.

Imaging and Spectroscopy of Natural Fluorophores in Pine Needles

PubMed Central

Williams, Nari

2018-01-01

Many plant tissues fluoresce due to the natural fluorophores present in cell walls or within the cell protoplast or lumen. While lignin and chlorophyll are well-known fluorophores, other components are less well characterized. Confocal fluorescence microscopy of fresh or fixed vibratome-cut sections of radiata pine needles revealed the presence of suberin, lignin, ferulate, and flavonoids associated with cell walls as well as several different extractive components and chlorophyll within tissues. Comparison of needles in different physiological states demonstrated the loss of chlorophyll in both chlorotic and necrotic needles. Necrotic needles showed a dramatic change in the fluorescence of extractives within mesophyll cells from ultraviolet (UV) excited weak blue fluorescence to blue excited strong green fluorescence associated with tissue browning. Comparisons were made among fluorophores in terms of optimal excitation, relative brightness compared to lignin, and the effect of pH of mounting medium. Fluorophores in cell walls and extractives in lumens were associated with blue or green emission, compared to the red emission of chlorophyll. Autofluorescence is, therefore, a useful method for comparing the histology of healthy and diseased needles without the need for multiple staining techniques, potentially aiding visual screening of host resistance and disease progression in needle tissue. PMID:29393922

Time-resolved and temperature tuneable measurements of fluorescent intensity using a smartphone fluorimeter.

PubMed

Hossain, Md Arafat; Canning, John; Yu, Zhikang; Ast, Sandra; Rutledge, Peter J; Wong, Joseph K-H; Jamalipour, Abbas; Crossley, Maxwell J

2017-05-30

A smartphone fluorimeter capable of time-based fluorescence intensity measurements at various temperatures is reported. Excitation is provided by an integrated UV LED (λ ex = 370 nm) and detection obtained using the in-built CMOS camera. A Peltier is integrated to allow measurements of the intensity over T = 10 to 40 °C. All components are controlled using a smartphone battery powered Arduino microcontroller and a customised Android application that allows sequential fluorescence imaging and quantification every δt = 4 seconds. The temperature dependence of fluorescence intensity for four emitters (rhodamine B, rhodamine 6G, 5,10,15,20-tetraphenylporphyrin and 6-(1,4,8,11-tetraazacyclotetradecane)2-ethyl-naphthalimide) are characterised. The normalised fluorescence intensity over time of the latter chemosensor dye complex in the presence of Zn 2+ is observed to accelerate with an increasing rate constant, k = 1.94 min -1 at T = 15 °C and k = 3.64 min -1 at T = 30 °C, approaching a factor of ∼2 with only a change in temperature of ΔT = 15 °C. Thermally tuning these twist and bend associated rates to optimise sensor approaches and device applications is proposed.

A unique charge-coupled device/xenon arc lamp based imaging system for the accurate detection and quantitation of multicolour fluorescence.

PubMed

Spibey, C A; Jackson, P; Herick, K

2001-03-01

In recent years the use of fluorescent dyes in biological applications has dramatically increased. The continual improvement in the capabilities of these fluorescent dyes demands increasingly sensitive detection systems that provide accurate quantitation over a wide linear dynamic range. In the field of proteomics, the detection, quantitation and identification of very low abundance proteins are of extreme importance in understanding cellular processes. Therefore, the instrumentation used to acquire an image of such samples, for spot picking and identification by mass spectrometry, must be sensitive enough to be able, not only, to maximise the sensitivity and dynamic range of the staining dyes but, as importantly, adapt to the ever changing portfolio of fluorescent dyes as they become available. Just as the available fluorescent probes are improving and evolving so are the users application requirements. Therefore, the instrumentation chosen must be flexible to address and adapt to those changing needs. As a result, a highly competitive market for the supply and production of such dyes and the instrumentation for their detection and quantitation have emerged. The instrumentation currently available is based on either laser/photomultiplier tube (PMT) scanning or lamp/charge-coupled device (CCD) based mechanisms. This review briefly discusses the advantages and disadvantages of both System types for fluorescence imaging, gives a technical overview of CCD technology and describes in detail a unique xenon/are lamp CCD based instrument, from PerkinElmer Life Sciences. The Wallac-1442 ARTHUR is unique in its ability to scan both large areas at high resolution and give accurate selectable excitation over the whole of the UV/visible range. It operates by filtering both the excitation and emission wavelengths, providing optimal and accurate measurement and quantitation of virtually any available dye and allows excellent spectral resolution between different fluorophores. This flexibility and excitation accuracy is key to multicolour applications and future adaptation of the instrument to address the application requirements and newly emerging dyes.

Luminescent single-walled carbon nanotube-sensitized europium nanoprobes for cellular imaging

PubMed Central

Avti, Pramod K; Sitharaman, Balaji

2012-01-01

Lanthanoid-based optical probes with excitation wavelengths in the ultra-violet (UV) range (300–325 nm) have been widely developed as imaging probes. Efficient cellular imaging requires that lanthanoid optical probes be excited at visible wavelengths, to avoid UV damage to cells. The efficacy of europium-catalyzed single-walled carbon nanotubes (Eu-SWCNTs), as visible nanoprobes for cellular imaging, is reported in this study. Confocal fluorescence microscopy images of breast cancer cells (SK-BR-3 and MCF-7) and normal cells (NIH 3T3), treated with Eu-SWCNT at 0.2 μg/mL concentration, showed bright red luminescence after excitation at 365 nm and 458 nm wavelengths. Cell viability analysis showed no cytotoxic effects after the incubation of cells with Eu-SWCNTs at this concentration. Eu-SWCNT uptake is via the endocytosis mechanism. Labeling efficiency, defined as the percentage of incubated cells that uptake Eu-SWCNT, was 95%–100% for all cell types. The average cellular uptake concentration was 6.68 ng Eu per cell. Intracellular localization was further corroborated by transmission electron microscopy and Raman microscopy. The results indicate that Eu-SWCNT shows potential as a novel cellular imaging probe, wherein SWCNT sensitizes Eu3+ ions to allow excitation at visible wavelengths, and stable time-resolved red emission. The ability to functionalize biomolecules on the exterior surface of Eu-SWCNT makes it an excellent candidate for targeted cellular imaging. PMID:22619533

Fluorescence properties of human teeth and dental calculus for clinical applications

NASA Astrophysics Data System (ADS)

Lee, Yong-Keun

2015-04-01

Fluorescent emission of human teeth and dental calculus is important for the esthetic rehabilitation of teeth, diagnosis of dental caries, and detection of dental calculus. The purposes of this review were to summarize the fluorescence and phosphorescence of human teeth by ambient ultraviolet (UV) light, to investigate the clinically relevant fluorescence measurement methods in dentistry, and to review the fluorescence of teeth and dental calculus by specific wavelength light. Dentine was three times more phosphorescent than enamel. When exposed to light sources containing UV components, the fluorescence of human teeth gives them the quality of vitality, and fluorescent emission with a peak of 440 nm is observed. Esthetic restorative materials should have fluorescence properties similar to those of natural teeth. Based on the fluorescence of teeth and restorative materials as determined with a spectrophotometer, a fluorescence parameter was defined. As to the fluorescence spectra by a specific wavelength, varied wavelengths were investigated for clinical applications, and several methods for the diagnosis of dental caries and the detection of dental calculus were developed. Since fluorescent properties of dental hard tissues have been used and would be expanded in diverse fields of clinical practice, these properties should be investigated further, embracing newly developed optical techniques.

Fluorescence properties of human teeth and dental calculus for clinical applications.

PubMed

Lee, Yong-Keun

2015-04-01

Fluorescent emission of human teeth and dental calculus is important for the esthetic rehabilitation of teeth, diagnosis of dental caries, and detection of dental calculus. The purposes of this review were to summarize the fluorescence and phosphorescence of human teeth by ambient ultraviolet (UV) light, to investigate the clinically relevant fluorescence measurement methods in dentistry, and to review the fluorescence of teeth and dental calculus by specific wavelength light. Dentine was three times more phosphorescent than enamel. When exposed to light sources containing UV components, the fluorescence of human teeth gives them the quality of vitality, and fluorescent emission with a peak of 440 nm is observed. Esthetic restorative materials should have fluorescence properties similar to those of natural teeth. Based on the fluorescence of teeth and restorative materials as determined with a spectrophotometer, a fluorescence parameter was defined. As to the fluorescence spectra by a specific wavelength, varied wavelengths were investigated for clinical applications, and several methods for the diagnosis of dental caries and the detection of dental calculus were developed. Since fluorescent properties of dental hard tissues have been used and would be expanded in diverse fields of clinical practice, these properties should be investigated further, embracing newly developed optical techniques.

Conjugation of (E)-5-[2-(Methoxycarbonyl)ethenyl]cytidine to hydrophilic microspheres: development of a mobile microscale UV light actinometer.

PubMed

Fang, Shiyue; Guan, Yousheng; Blatchley, Ernest R; Shen, Chengyue; Bergstrom, Donald E

2008-03-01

( E)-5-[2-(Methoxycarbonyl)ethenyl]cytidine was biotinylated through a diisopropylsilylacetal linkage and attached to the surface of hydrophilic streptavidin-coated microspheres through the high-affinity noncovalent interaction between biotin and streptavidin. The functionalized microspheres form a stable suspension in water. Upon UV irradiation, the nonfluorescent ( E)-5-[2-(methoxycarbonyl)ethenyl]cytidine on the microspheres undergoes photocyclization to produce highly fluorescent 3-beta-D-ribofuranosyl-2,7-dioxopyrido[2,3-d]pyrimidine. The fluorescence intensity of the microspheres can be correlated to the particle-specific UV doses applied at different suspension concentrations. The microspheres allow one to measure the UV dose (fluence) distribution in high-throughput water disinfection systems.

Light-induced fluorescence changes in Phycomyces: evidence for blue light-receptor associated flavo-semiquinones.

PubMed

Galland, Paul; Tölle, Nadja

2003-10-01

Light-induced fluorescence changes (LIFCs) were detected in sporangiophores of the blue-light-sensitive fungus Phycomyces blakesleeanus (Burgeff). The LIFCs can be utilized as a spectrophotometric assay for blue-light photoreceptors and for the in vivo characterization of their photochemical primary reactions. Blue-light irradiation of sporangiophores elicited a transient decrease and subsequent regeneration of flavin-like fluorescence emission at 525 nm. The signals recovered in darkness in about 120 min. In contrast to blue light, near-UV (370 nm) caused an increase in the fluorescence emission at 525 nm. Because the LIFCs were altered in a light-insensitive madC mutant with a defective photoreceptor, the fluorescence changes must be associated with early photochemical events of the transduction chain. Action spectra for the fluorescence changes at 525 nm showed major peaks near 470 and 600 nm. Double-pulse experiments involving two consecutive pulses of either blue and near-UV, blue and red, or near-UV and red showed that the responses depended on the sequence in which the different wavelengths were applied. The results indicate a blue-light receptor with intermediates in the near-UV, blue and red spectral regions. We explain the results in the framework of a general model, in which the three redox states of the flavin photoreceptor, the oxidized flavin (Fl), the flavo-semiquinone (FlH*), and the flavo-hydroquinone (FlH2) are each acting as chromophores with their own characteristic photochemical primary reactions. These consist of the photoreduction of the oxidized flavin generating semiquinone, the photoreduction of the semiquinone generating hydroquinone, and the photooxidation of the flavo-hydroquinone regenerating the pool of oxidized flavins. The proposed mechanism represents a photocycle in which two antagonistic photoreceptor forms, Fl and FlH2, determine the pool size of the biological effector molecule, the flavo-semiquinone. The redox changes that are associated with the photocycle are maintained by redox partners, pterins, that function in the near-UV as secondary chromophores.

Lettuce flavonoids screening and phenotyping by chlorophyll fluorescence excitation ratio.

PubMed

Zivcak, Marek; Brückova, Klaudia; Sytar, Oksana; Brestic, Marian; Olsovska, Katarina; Allakhverdiev, Suleyman I

2017-06-01

Environmentally induced variation and the genotypic differences in flavonoid and phenolic content in lettuce can be reliably detected using the appropriate parameters derived from the records of rapid non-invasive fluorescence technique. The chlorophyll fluorescence excitation ratio method was designed as a rapid and non-invasive tool to estimate the content of UV-absorbing phenolic compounds in plants. Using this technique, we have assessed the dynamics of accumulation of flavonoids related to developmental changes and environmental effects. Moreover, we have tested appropriateness of the method to identify the genotypic differences and fluctuations in total phenolics and flavonoid content in lettuce. Six green and two red genotypes of lettuce (Lactuca sativa L.) grown in pots were exposed to two different environments for 50 days: direct sunlight (UV-exposed) and greenhouse conditions (low UV). The indices based on the measurements of chlorophyll fluorescence after red, green and UV excitation indicated increase of the content of UV-absorbing compounds and anthocyanins in the epidermis of lettuce leaves. In similar, the biochemical analyses performed at the end of the experiment confirmed significantly higher total phenolic and flavonoid content in lettuce plants exposed to direct sun compared to greenhouse conditions and in red compared to green genotypes. As the correlation between the standard fluorescence indices and the biochemical records was negatively influenced by the presence of red genotypes, we proposed the use of a new parameter named Modified Flavonoid Index (MFI) taking into an account both absorbance changes due to flavonol and anthocyanin content, for which the correlation with flavonoid and phenolic content was relatively good. Thus, our results confirmed that the fluorescence excitation ratio method is useful for identifying the major differences in phenolic and flavonoid content in lettuce plants and it can be used for high-throughput pre-screening and phenotyping of leafy vegetables in research and breeding applications towards improvement of vegetable health effects.

Facile preparation of fluorescent layered double hydroxide polymeric composites through the photo-induced surface-initiated controlled living polymerization

NASA Astrophysics Data System (ADS)

Chen, Junyu; Liu, Meiying; Huang, Qiang; Jiang, Ruming; Huang, Hongye; Deng, Fengjie; Wen, Yuanqing; Tian, Jianwen; Zhang, Xiaoyong; Wei, Yen

2018-05-01

(Zn/Al) layered double hydroxide (LDH) based fluorescence probes have been facilely fabricated via photo-induced surface-initiated reversible addition-fragmentation chain transfer (RAFT) polymerization, which demonstrated green fluorescence, good biocompatibility and excellent dispersion performance in aqueous solution. The as prepared (Zn/Al)LDH polymeric composites were modified with 2-methacryloyloxyethyl phosphorylcholine (MPC), acrylic acid (AA) and diacroloyl-fluorescein (Ac-Fl). Among them, the comonomers MPC and AA were used to endow their water dispersibility, biocompatibility and potential drug carriers, while the Ac-Fl was served both as the fluorescence signal and photocatalyst for RAFT polymerization. A series of characterization methods, including 1H nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, transmission electronic microscopy, thermogravimetric analyses, X-ray photoelectron spectroscopy were employed to conform the successful of surface modification of LDH through photo-induced surface-initiated RAFT polymerization. Besides, UV-vis absorption spectra and fluorescence spectra were adopted to evaluate the optical characteristics of as prepared (Zn/Al)LDH-co-Poly(MPC-AA-Fl) composites, which exhibited high intense green fluorescence. Furthermore, the endocytosis behavior indicates that (Zn/Al)LDH-co-Poly(MPC-AA-Fl) composites could be potentially used in cell imaging and even drug delivery application for their excellent biocompatibility and all advantages described above.

Ambient temperature effects on broadband UV-B measurements using fluorescent phosphor (MgWO4)-based detectors

NASA Technical Reports Server (NTRS)

Dichter, Bronislaw K.; Beaubien, David J.; Beaubien, Arthur F.

1994-01-01

Results of field tests on a group of broadband UV-B pyranometers are presented. A brief description of the instrument is given. The effects of ambient temperature on thermally unregulated fluorescent phosphor (Robertson type) meters are presented and compared with the performance of thermally stabilized instruments. Means for correcting data from thermally unregulated instruments, where the prevailing ambient temperatures are known, are outlined.

Preliminary investigation of intrinsic UV fluorescence spectroscopic changes associated with proteolytic digestion of bovine articular cartilage

NASA Astrophysics Data System (ADS)

Lewis, William; Padilla-Martinez, Juan-Pablo; Ortega-Martinez, Antonio; Franco, Walfre

2016-03-01

Degradation and destruction of articular cartilage is the etiology of osteoarthritis (OA), an entity second only to cardiovascular disease as a cause of disability in the United States. Joint mechanics and cartilage biochemistry are believed to play a role in OA; an optical tool to detect structural and chemical changes in articular cartilage might offer benefit for its early detection and treatment. The objective of the present study was to identify the spectral changes in intrinsic ultraviolet (UV) fluorescence of cartilage that occur after proteolytic digestion of cartilage. Bovine articular cartilage samples were incubated in varying concentrations of collagenase ranging from 10ug/mL up to 5mg/mL for 18 hours at 37°C, a model of OA. Pre- and post-incubation measurements were taken of the UV excitation-emission spectrum of each cartilage sample. Mechanical tests were performed to determine the pre- and post-digestion force/displacement ratio associated with indentation of each sample. Spectral changes in intrinsic cartilage fluorescence and stiffness of the cartilage were associated with proteolytic digestion. In particular, changes in the relative intensity of fluorescence peaks associated with pentosidine crosslinks (330 nm excitation, 390 nm emission) and tryptophan (290 nm excitation, 340 nm emission) were found to correlate with different degrees of cartilage digestion and cartilage stiffness. In principle, it may be possible to use UV fluorescence spectral data for early detection of damage to articular cartilage, and as a surrogate measure for cartilage stiffness.

Fluorescence spectroscopy for non-invasive measurement of mechanical stiffness after photo-crosslinking of rabbit cornea

NASA Astrophysics Data System (ADS)

Williams, Maura; Lewis, William; Ortega-Martinez, Antonio; Franco, Walfre

2017-02-01

Background and Objectives: Keratoconus is a disease characterized by progressive steepening and thinning of the cornea, altering visual acuity and sometimes potentiating the need for corneal transplant if the disease progresses.1-3 Corneal crosslinking, a procedure that uses topical riboflavin and UV light to increase the stiffness of the cornea through the creation of collagen crosslinks was recently approved by the FDA for use in the U.S. The objective of the present study was to investigate whether endogenous collagen fluorescence changes following treatment can be correlated to alterations in the stiffness of the cornea, thereby guiding treatment parameters. Study Design and Results: 78 ex-vivo rabbit eyes divided into three groups: riboflavin solution plus UV irradiation, dextran solution plus UV irradiation, and riboflavin solution only. An additional group of eyes received no treatment. The epithelium was removed from each sample and topical riboflavin was applied. Eyes were irradiated with a 365 nm black ray UV lamp for various treatment times, ranging from half the clinical treatment time to three times the length. Mechanical testing was performed to determine the force/displacement relationship for the various treatment times. Fluorescence spectral changes following treatment corresponded with changes in stiffness. In particular, a decrease in the value of fluorescence intensity at 290/340 nm excitation/emission wavelengths corresponded to an increase in corneal stiffness following treatment. It may be possible to use fluorescence spectral changes of endogenous corneal crosslinks to evaluate mechanical stiffness changes non-invasively.

Evolution of molecular weight and fluorescence of effluent organic matter (EfOM) during oxidation processes revealed by advanced spectrographic and chromatographic tools.

PubMed

Chen, Zhiqiang; Li, Mo; Wen, Qinxue; Ren, Nanqi

2017-11-01

Effluent organic matter (EfOM) is an emerging concern to receiving aquatic environment due to its refractory property. The degradation of EfOM in ozonation and other two advanced oxidation processes (AOPs), UV/H 2 O 2 and UV/persulfate (PS), was investigated in this study. Fluorescence spectra coupled with parallel factor analysis (PARAFAC) and two-dimensional correlation gel permeation chromatography (2D-GPC) were used to track the evolution of EfOM during each oxidation process. Results showed that the degradation of EfOM indicated by dissolved organic carbon (DOC), UV 254 and fluorescence components, fitted well with pseudo-first-order kinetic model during the oxidation processes. Ozonation showed higher degradation efficiency than AOPs, while UV/PS was more effective than UV/H 2 O 2 with equimolar oxidants dosage. Ozone and SO· 4 - were more reactive with terrestrial humic-like substances, while hydroxyl radical preferentially reacted with protein-like substances. Organic molecules with higher molecular weight (MW) were susceptible to ozone or radicals. Ozonation could transform higher MW (MW of 3510 and 575) organic matters into lower MW organic matters (MW of 294), while reductions of all the organics were observed in both AOPs. Due to the higher reaction rates between ozone and EfOM, ozonation maybe serve as a pre-treatment for AOPs to reduce the radical and energy consumption and improve mineralization of EfOM by AOPs. The decline in DOC, UV 254 , fluorescence and reduction in oxidants increased with the increase of oxidants dosage, and linear correlations among them were found during the ozonation and AOPs. Copyright © 2017. Published by Elsevier Ltd.

Photoswitchable non-fluorescent thermochromic dye-nanoparticle hybrid probes

PubMed Central

Harrington, Walter N.; Haji, Mwafaq R.; Galanzha, Ekaterina I.; Nedosekin, Dmitry A.; Nima, Zeid A.; Watanabe, Fumiya; Ghosh, Anindya; Biris, Alexandru S.; Zharov, Vladimir P.

2016-01-01

Photoswitchable fluorescent proteins with controllable light–dark states and spectral shifts in emission in response to light have led to breakthroughs in the study of cell biology. Nevertheless, conventional photoswitching is not applicable for weakly fluorescent proteins and requires UV light with low depth penetration in bio-tissue. Here we introduce a novel concept of photoswitchable hybrid probes consisting of thermochromic dye and absorbing nanoparticles, in which temperature-sensitive light–dark states and spectral shifts in absorption can be switched through controllable photothermal heating of doped nanoparticles. The proof-of-concept is demonstrated through the use of two different types of temperature-sensitive dyes doped with magnetic nanoparticles and reversibly photoswitched by a near-infrared laser. Photoacoustic imaging revealed the high contrast of these probes, which is sufficient for their visualization in cells and deep tissue. Our results suggest that these new photoswitchable multicolour probes can be used for multimodal cellular diagnostics and potentially for magnetic and photothermal therapy. PMID:27824110

Chemical probes for analysis of carbonylated proteins: a review

PubMed Central

Yan, Liang-Jun; Forster, Michael J.

2010-01-01

Protein carbonylation is a major form of protein oxidation and is widely used as an indicator of oxidative stress. Carbonyl groups do not have distinguishing UV or visible, spectrophotometric absorbance/fluorescence characteristics and thus their detection and quantification can only be achieved using specific chemical probes. In this paper, we review the advantages and disadvantages of several chemical probes that have been and are still being used for protein carbonyl analysis. These probes include 2, 4-dinitrophenylhydazine (DNPH), tritiated sodium borohydride ([3H]NaBH4), biotin-containing probes, and fluorescence probes. As our discussions lean toward gel-based approaches, utilizations of these probes in 2D gel-based proteomic analysis of carbonylated proteins are illustrated where applicable. Analysis of carbonylated proteins by ELISA, immunofluorescent imaging, near infrared fluorescence detection, and gel-free proteomic approaches are also discussed where appropriate. Additionally, potential applications of blue native gel electrophoresis as a tool for first dimensional separation in 2D gel-based analysis of carbonylated proteins are discussed as well. PMID:20732835

Cloning of genes involved in the biosynthesis of pseudobactin, a high-affinity iron transport agent of a plant growth-promoting Pseudomonas strain.

PubMed Central

Moores, J C; Magazin, M; Ditta, G S; Leong, J

1984-01-01

A gene bank of DNA from plant growth-promoting Pseudomonas sp. strain B10 was constructed using the broad host-range conjugative cosmid pLAFR1. The recombinant cosmids contained insert DNA averaging 21.5 kilobase pairs in length. Nonfluorescent mutants of Pseudomonas sp. strain B10 were obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, or UV light and were defective in the biosynthesis of its yellow-green, fluorescent siderophore (microbial iron transport agent) pseudobactin. No yellow-green, fluorescent mutants defective in the production of pseudobactin were identified. Nonfluorescent mutants were individually complemented by mating the gene bank en masse and identifying fluorescent transconjugants. Eight recombinant cosmids were sufficient to complement 154 nonfluorescent mutants. The pattern of complementation suggests that a minimum of 12 genes arranged in four gene clusters is required for the biosynthesis of pseudobactin. This minimum number of genes seems reasonable considering the structural complexity of pseudobactin. Images PMID:6690426

Forensic photography. Ultraviolet imaging of wounds on skin.

PubMed

Barsley, R E; West, M H; Fair, J A

1990-12-01

The use of ultraviolet light (UVL) to study and document patterned injuries on human skin has opened a new frontier for law enforcement. This article discusses the photographic techniques involved in reflective and fluorescent UVL. Documentation of skin wounds via still photography and dynamic video photographic techniques, which utilize various methods of UV illumination, are covered. Techniques important for courtroom presentation of evidence gathered from lacerations, contusions, abrasions, and bite marks are presented through case studies and controlled experiments. Such injuries are common sequelae in the crimes of child abuse, rape, and assault.

UV reactor flow visualization and mixing quantification using three-dimensional laser-induced fluorescence.

PubMed

Gandhi, Varun; Roberts, Philip J W; Stoesser, Thorsten; Wright, Harold; Kim, Jae-Hong

2011-07-01

Three-dimensional laser-induced fluorescence (3DLIF) was applied to visualize and quantitatively analyze mixing in a lab-scale UV reactor consisting of one lamp sleeve placed perpendicular to flow. The recirculation zone and the von Karman vortex shedding that commonly occur in flows around bluff bodies were successfully visualized. Multiple flow paths were analyzed by injecting the dye at various heights with respect to the lamp sleeve. A major difference in these pathways was the amount of dye that traveled close to the sleeve, i.e., a zone of higher residence time and higher UV exposure. Paths away from the center height had higher velocities and hence minimal influence by the presence of sleeve. Approach length was also characterized in order to increase the probability of microbes entering the region around the UV lamp. The 3DLIF technique developed in this study is expected to provide new insight on UV dose delivery useful for the design and optimization of UV reactors. Copyright © 2011 Elsevier Ltd. All rights reserved.

Self-assembly of BODIPY based pH-sensitive near-infrared polymeric micelles for drug controlled delivery and fluorescence imaging applications

NASA Astrophysics Data System (ADS)

Liu, Xiaodong; Chen, Bizheng; Li, Xiaojun; Zhang, Lifen; Xu, Yujie; Liu, Zhuang; Cheng, Zhenping; Zhu, Xiulin

2015-10-01

Responsive block copolymer micelles emerging as promising imaging and drug delivery systems show high stability and on-demand drug release activities. Herein, we developed self-assembled pH-responsive NIR emission micelles entrapped with doxorubicin (DOX) within the cores by the electrostatic interactions for fluorescence imaging and chemotherapy applications. The block copolymer, poly(methacrylic acid)-block-poly[(poly(ethylene glycol) methyl ether methacrylate)-co-boron dipyrromethene derivatives] (PMAA-b-P(PEGMA-co-BODIPY)), was synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, and the molecular weight distribution of this copolymer was narrow (Mw/Mn = 1.31). The NIR fluorescence enhancement induced by the phenol/phenolate interconversion equilibrium works as a switch in response to the intracellular pH fluctuations. DOX-loaded PMAA-b-P(PEGMA-co-BODIPY) micelles can detect the physiological pH fluctuations with a pKa near physiological conditions (~7.52), and showed pH-responsive collapse and an obvious acid promoted anticancer drug release behavior (over 58.8-62.8% in 10 h). Real-time imaging of intracellular pH variations was performed and a significant chemotherapy effect was demonstrated against HeLa cells.Responsive block copolymer micelles emerging as promising imaging and drug delivery systems show high stability and on-demand drug release activities. Herein, we developed self-assembled pH-responsive NIR emission micelles entrapped with doxorubicin (DOX) within the cores by the electrostatic interactions for fluorescence imaging and chemotherapy applications. The block copolymer, poly(methacrylic acid)-block-poly[(poly(ethylene glycol) methyl ether methacrylate)-co-boron dipyrromethene derivatives] (PMAA-b-P(PEGMA-co-BODIPY)), was synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, and the molecular weight distribution of this copolymer was narrow (Mw/Mn = 1.31). The NIR fluorescence enhancement induced by the phenol/phenolate interconversion equilibrium works as a switch in response to the intracellular pH fluctuations. DOX-loaded PMAA-b-P(PEGMA-co-BODIPY) micelles can detect the physiological pH fluctuations with a pKa near physiological conditions (~7.52), and showed pH-responsive collapse and an obvious acid promoted anticancer drug release behavior (over 58.8-62.8% in 10 h). Real-time imaging of intracellular pH variations was performed and a significant chemotherapy effect was demonstrated against HeLa cells. Electronic supplementary information (ESI) available: GPC, UV/vis, fluorescence, and MTT data of the as-prepared polymers; 1H NMR, 13C NMR, HRMS and FT-IR of organic molecules and polymers. See DOI: 10.1039/c5nr04655f

Characterization of a fluorescent hydrogel synthesized using chitosan, polyvinyl alcohol and 9-anthraldehyde for the selective detection and discrimination of trace Fe3+ and Fe2+ in water for live-cell imaging.

PubMed

Maity, Santu; Parshi, Nira; Prodhan, Chandraday; Chaudhuri, Keya; Ganguly, Jhuma

2018-08-01

A three-dimensional fluorescent hydrogel based on chitosan, polyvinyl alcohol and 9-anthraldehyde (ChPA) has been successfully designed and synthesized for the selective detection and discrimination of Fe 3+ and Fe 2+ in aqueous environment. The unique characteristics of ChPA has been confirmed by the Fourier-transform infrared spectroscopy (FTIR), rheological measurement, scanning electron microscopy (SEM), thermogravimetry and differential thermogravimetry (TG-DTG), ultraviolet-visible spectroscopy (UV-vis), fluorescence studies, transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDX), x-ray diffraction (XRD) and dynamic light scattering (DLS). The emission intensity at 516 nm of the hydrogel has been enhanced remarkably with the addition of Fe 3+ due to the inhibition of the photoinduced electron transfer (PET) process. However, it gets strongly quenched in the case of Fe 2+ owing to chelation enhanced quenching (CHEQ). The probe (ChPA) causes no significant change in the fluorescence and becomes highly specific and sensitive towards Fe 3+ and Fe 2+ compared to other interfering heavy and transition metal ions (HTM). The detection limits of the sensor for the Fe 3+ and Fe 2+ are 0.124 nM and 0.138 nM, respectively. The probe is also promising as a selective sensor for the Fe 3+ and Fe 2+ in the fluorescence imaging of living cells. Thus, such a probe opens up new opportunities to improve the chitosan based fluorescent chemosensor having biocompatibility, biodegradability, sufficient thermal stability and stability in a wide pH range. Copyright © 2018 Elsevier Ltd. All rights reserved.

UV-Light Exposure of Insulin: Pharmaceutical Implications upon Covalent Insulin Dityrosine Dimerization and Disulphide Bond Photolysis

PubMed Central

Correia, Manuel; Neves-Petersen, Maria Teresa; Jeppesen, Per Bendix; Gregersen, Søren; Petersen, Steffen B.

2012-01-01

In this work we report the effects of continuous UV-light (276 nm, ∼2.20 W.m−2) excitation of human insulin on its absorption and fluorescence properties, structure and functionality. Continuous UV-excitation of the peptide hormone in solution leads to the progressive formation of tyrosine photo-product dityrosine, formed upon tyrosine radical cross-linkage. Absorbance, fluorescence emission and excitation data confirm dityrosine formation, leading to covalent insulin dimerization. Furthermore, UV-excitation of insulin induces disulphide bridge breakage. Near- and far-UV-CD spectroscopy shows that UV-excitation of insulin induces secondary and tertiary structure losses. In native insulin, the A and B chains are held together by two disulphide bridges. Disruption of either of these bonds is likely to affect insulin’s structure. The UV-light induced structural changes impair its antibody binding capability and in vitro hormonal function. After 1.5 and 3.5 h of 276 nm excitation there is a 33.7% and 62.1% decrease in concentration of insulin recognized by guinea pig anti-insulin antibodies, respectively. Glucose uptake by human skeletal muscle cells decreases 61.7% when the cells are incubated with pre UV-illuminated insulin during 1.5 h. The observations presented in this work highlight the importance of protecting insulin and other drugs from UV-light exposure, which is of outmost relevance to the pharmaceutical industry. Several drug formulations containing insulin in hexameric, dimeric and monomeric forms can be exposed to natural and artificial UV-light during their production, packaging, storage or administration phases. We can estimate that direct long-term exposure of insulin to sunlight and common light sources for indoors lighting and UV-sterilization in industries can be sufficient to induce irreversible changes to human insulin structure. Routine fluorescence and absorption measurements in laboratory experiments may also induce changes in protein structure. Structural damage includes insulin dimerization via dityrosine cross-linking or disulphide bond disruption, which affects the hormone’s structure and bioactivity. PMID:23227203

UV-light exposure of insulin: pharmaceutical implications upon covalent insulin dityrosine dimerization and disulphide bond photolysis.

PubMed

Correia, Manuel; Neves-Petersen, Maria Teresa; Jeppesen, Per Bendix; Gregersen, Søren; Petersen, Steffen B

2012-01-01

In this work we report the effects of continuous UV-light (276 nm, ~2.20 W.m(-2)) excitation of human insulin on its absorption and fluorescence properties, structure and functionality. Continuous UV-excitation of the peptide hormone in solution leads to the progressive formation of tyrosine photo-product dityrosine, formed upon tyrosine radical cross-linkage. Absorbance, fluorescence emission and excitation data confirm dityrosine formation, leading to covalent insulin dimerization. Furthermore, UV-excitation of insulin induces disulphide bridge breakage. Near- and far-UV-CD spectroscopy shows that UV-excitation of insulin induces secondary and tertiary structure losses. In native insulin, the A and B chains are held together by two disulphide bridges. Disruption of either of these bonds is likely to affect insulin's structure. The UV-light induced structural changes impair its antibody binding capability and in vitro hormonal function. After 1.5 and 3.5 h of 276 nm excitation there is a 33.7% and 62.1% decrease in concentration of insulin recognized by guinea pig anti-insulin antibodies, respectively. Glucose uptake by human skeletal muscle cells decreases 61.7% when the cells are incubated with pre UV-illuminated insulin during 1.5 h. The observations presented in this work highlight the importance of protecting insulin and other drugs from UV-light exposure, which is of outmost relevance to the pharmaceutical industry. Several drug formulations containing insulin in hexameric, dimeric and monomeric forms can be exposed to natural and artificial UV-light during their production, packaging, storage or administration phases. We can estimate that direct long-term exposure of insulin to sunlight and common light sources for indoors lighting and UV-sterilization in industries can be sufficient to induce irreversible changes to human insulin structure. Routine fluorescence and absorption measurements in laboratory experiments may also induce changes in protein structure. Structural damage includes insulin dimerization via dityrosine cross-linking or disulphide bond disruption, which affects the hormone's structure and bioactivity.

Diurnal changes in epidermal UV transmittance of plants in naturally high UV environments.

PubMed

Barnes, Paul W; Flint, Stephan D; Slusser, James R; Gao, Wei; Ryel, Ronald J

2008-06-01

Studies were conducted on three herbaceous plant species growing in naturally high solar UV environments in the subalpine of Mauna Kea, Hawaii, USA, to determine if diurnal changes in epidermal UV transmittance (T(UV)) occur in these species, and to test whether manipulation of the solar radiation regime could alter these diurnal patterns. Additional field studies were conducted at Logan, Utah, USA, to determine if solar UV was causing diurnal T(UV) changes and to evaluate the relationship between diurnal changes in T(UV) and UV-absorbing pigments. Under clear skies, T(UV), as measured with a UV-A-pulse amplitude modulation fluorometer for leaves of Verbascum thapsus and Oenothera stricta growing in native soils and Vicia faba growing in pots, was highest at predawn and sunset and lowest at midday. These patterns in T(UV) closely tracked diurnal changes in solar radiation and were the result of correlated changes in fluorescence induced by UV-A and blue radiation but not photochemical efficiency (F(v)/F(m)) or initial fluorescence yield (F(o)). The magnitude of the midday reduction in T(UV) was greater for young leaves than for older leaves of Verbascum. Imposition of artificial shade eliminated the diurnal changes in T(UV) in Verbascum, but reduction in solar UV had no effect on diurnal T(UV) changes in Vicia. In Vicia, the diurnal changes in T(UV) occurred without detectable changes in the concentration of whole-leaf UV-absorbing compounds. Results suggest that plants actively control diurnal changes in UV shielding, and these changes occur in response to signals other than solar UV; however, the underlying mechanisms responsible for rapid changes in T(UV) remain unclear.

The HST-pNFL program: Mapping the Fluorescent Emission of Galactic Outflows

NASA Astrophysics Data System (ADS)

Heckman, Timothy

2017-08-01

Galactic outflows associated with star formation are believed to play a crucial role in the evolution of galaxies and the IGM. Most of our knowledge about outflows has come from down-the-barrel UV absorption spectroscopy of star-forming galaxies. However, absorption-line data alone provide only indirect information about the radial structure of the gas flows, which introduces large systematic uncertainties in some of the most important quantities, such as the outflow rate, the mass loading factor, and the momentum, metal, and energy fluxes. Recent spectroscopic observations of star-forming galaxies with large (projected physical) apertures have revealed non-resonant (fluorescent) emission in the UV, e.g., FeII* and SiII*, that can be naturally produced by spatially extended emission from the same outflowing material traced in absorption. Encouraged by the most recent observations of FeII* emission by the SDSS-IV/eBOSS survey (Zhu et al. 2015), we propose a pilot program to use narrow-band filter UVIS F280N images to map the extended FeII* 2626 and 2613 fluorescent emission in a carefully-chosen sample of 4 starburst galaxies at z=0.065, and COS G130M to obtain down-the- barrel spectra for SiII absorption and SiII* emission. This HST pilot program can provide unique information about the spatial structure of galactic outflows and can potentially lead to a revolution in our understanding of outflow physics and its impact on galaxies and the IGM.

Silicon nanocrystals as handy biomarkers

NASA Astrophysics Data System (ADS)

Fujioka, Kouki; Hoshino, Akiyoshi; Manabe, Noriyoshi; Futamura, Yasuhiro; Tilley, Richard; Yamamoto, Kenji

2007-02-01

Quantum dots (QDs) have brighter and longer fluorescence than organic dyes. Therefore, QDs can be applied to biotechnology, and have capability to be applied to medical technology. Currently, among the several types of QDs, CdSe with a ZnS shell is one of the most popular QDs to be used in biological experiments. However, when the CdSe QDs were applied to clinical technology, potential toxicological problems due to CdSe core should be considered. To eliminate the problem, silicon nanocrystals, which have the potential of biocompatibility, could be a candidate of alternate probes. Silicon nanocrystals have been synthesized using several techniques such as aerosol, electrochemical etching, laser pyrolysis, plasma deposition, and colloids. Recently, the silicon nanocrystals were reported to be synthesized in inverse micelles and also stabilized with 1-heptene or allylamine capping. Blue fluorescence of the nanocrystals was observed when excited with a UV light. The nanocrystals covered with 1-heptene are hydrophobic, whereas the ones covered with allylamine are hydrophilic. To test the stability in cytosol, the water-soluble nanocrystals covered with allylamine were examined with a Hela cell incorporation experiment. Bright blue fluorescence of the nanocrystals was detected in the cytosol when excited with a UV light, implying that the nanocrystals were able to be applied to biological imaging. In order to expand the application range, we synthesized and compared a series of silicon nanocrystals, which have variable surface modification, such as alkyl group, alcohol group, and odorant molecules. This study will provide a wider range of optoelectronic applications and bioimaging technology.

Direct fluorescent labeling for efficient biological assessment of inhalable particles.

PubMed

Poudel, Bijay Kumar; Park, Jae Hong; Lim, Jiseok; Byeon, Jeong Hoon

2017-10-01

Labeling of aerosol particles with a radioactive, magnetic, or optical tracer has been employed to confirm particle localization in cell compartments, which has provided useful evidence for correlating toxic effects of inhaled particles. However, labeling requires several physicochemical steps to identify functionalities of the inner or outer surfaces of particles, and moreover, these steps can cause changes in size, surface charge, and bioactivity of the particles, resulting in misinterpretations regarding their toxic effects. This study addresses this challenging issue with a goal of introducing an efficient strategy for constantly supplying labeled aerosol particles in a single-pass configuration without any pre- or post-physicochemical batch treatments of aerosol particles. Carbon black (CB, simulating combustion-generated soot) or calcium carbonate (CC, simulating brake-wear fragments) particles were constantly produced via spark ablation or bubble bursting, respectively. These minute particles were incorporated with fluorescein isothiocyanate-poly(ethylene glycol) 2-aminoethyl ether acetic acid solution at the orifice of a collison atomizer to fabricate hybrid droplets. The droplets successively entered a diffusion dryer containing 254-nm UV irradiation; therefore, the droplets were dynamically stiffened by UV to form fluorescent probes on particles during solvent extraction in the dryer. Particle size distributions, morphologies, and surface charges before and after labeling were measured to confirm fluorescence labeling without significant changes in the properties. In vitro assays, including confocal imaging, were conducted to confirm the feasibility of the labeling approach without inducing significant differences in bioactivity compared with untreated CB or CC particles.

Characterization and Imaging of Antibody-Coated Gold Nanoparticles for Targeted Treatment of Microbial Keratitis

NASA Astrophysics Data System (ADS)

Mahan, Matthew

Microbial keratitis (MK) is an infection of the cornea by pathogenic organisms that causes inflammation and irritation. It can lead to full or partial blindness if left untreated. Current clinical treatment methods rely on high frequency application of topical drugs which are subject to the issues of patient compliance and microbial resistance. In this work, gold nanoparticles (AuNP) were proposed as an alternative treatment method in light-based therapies. Particle formulation methods were investigated and assessed using transmission electron microscopy (TEM) and ultraviolet/visible spectroscopy (UV-Vis). AuNP of 20 nm diameter were used as platforms to attach monoclonal antibodies anti-FLAG or anti-F1 to enhance their cell-targeting ability as well as polyethylene glycol to reduce non-specific binding and protein adsorption. These functionalized particles were qualitatively assessed using UV-Vis. The antibody-functionalized AuNP were then assessed for their ability to attach directly to Pseudomonas aeruginosa, expressing FLAG peptide, or Aspergillus fumigatus, expressing the F1 receptor. Attachment was imaged using dark field microscopy, transmission electron microscopy, and fluorescence microscopy.

Renilla luciferase- Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals.

PubMed

Wang, Y; Yu, Y A; Shabahang, S; Wang, G; Szalay, A A

2002-10-01

Light-emitting reporter proteins play an increasing role in the study of gene expression in vitro and in vivo. Here we present a ruc-gfp fusion gene construct generated by fusing a cDNA for Renilla luciferase (ruc) in-frame with a cDNA encoding the "humanized" GFP (gfp) from Aequorea. A plasmid containing the fusion gene construct was successfully transformed into, and expressed in, mammalian cells. The transformed cells exhibited both Renilla luciferase activity in the presence of coelenterazine and GFP fluorescence upon excitation with UV light. Spectrofluorometry of cells containing the Ruc-GFP fusion protein, in the absence of wavelengths capable of exciting GFP fluorescence but in the presence of the luciferase substrate, coelenterazine, showed an emission spectrum with two peaks at 475 nm and 508 nm. These two peaks correspond to the emission maximum of Renilla luciferase at 475 nm and that of GFP at 508 nm. The peak at 508 nm generated in the presence of coelenterazine alone (without UV excitation) is the result of intramolecular energy transfer from Renilla luciferase to Aequorea GFP. Southern analysis of genomic DNA purified from transformed Chinese hamster ovary (CHO) cells and fluorescence in situ hybridization (FISH) to metaphase chromosomes confirmed the integration of the ruc-gfp fusion gene on a single chromosome. The bifunctional Ruc-GFP fusion protein allows the detection of gene expression at the single-cell level based on green fluorescence, and in a group of cells based on luminescence emission. Furthermore, animal experiments revealed that light emission from the Ruc-GFP fusion protein can be detected externally in the organs or tissues of live animals bearing the gene construct.

UV-B radiation and photosynthetic irradiance acclimate eggplant for outdoor exposure

NASA Technical Reports Server (NTRS)

Latimer, J. G.; Mitchell, C. A.; Mitchell, G. A.

1987-01-01

Treatment of greenhouse-grown eggplant (Solanum melongena L. var. esculentum Nees. 'Burpee's Black Beauty') seedlings with supplemental photosynthetically active radiation from cool-white fluorescent lamps increased growth of plants subsequently transferred outdoors relative to growth of plants that received no supplemental radiation or were shaded to 45% of solar irradiation in the greenhouse before transfer outdoors. Eggplant seedlings transferred outdoors were placed under plastic tarps either to provide relative protection from solar ultraviolet-B (UV-B) radiation (280-315 nm) using Mylar film or to allow exposure to UV-B using cellulose acetate. Protection of seedlings from UV-B radiation resulted in greater leaf expansion than for UV-B-exposed seedlings, but no change in leaf or shoot dry weight occurred after 9 days of treatment. Specific leaf weight increased in response to UV-B exposure outdoors. Exposure of eggplant to UV-B radiation from fluorescent sunlamps in the greenhouse also decreased leaf expansion and leaf and shoot dry weight gain after 5 days of treatment. However, there were no differences in leaf or shoot dry weight relative to control plants after 12 days of UV-B treatment, indicating that UV-B treated plants had acclimated to the treatment and actually had caught up with non-UV-B-irradiated plants in terms of growth.

UV-B radiation and photosynthetic irradiance acclimate eggplant for outdoor exposure.

PubMed

Latimer, J G; Mitchell, C A; Mitchell, G A

1987-06-01

Treatment of greenhouse-grown eggplant (Solanum melongena L. var. esculentum Nees. 'Burpee's Black Beauty') seedlings with supplemental photosynthetically active radiation from cool-white fluorescent lamps increased growth of plants subsequently transferred outdoors relative to growth of plants that received no supplemental radiation or were shaded to 45% of solar irradiation in the greenhouse before transfer outdoors. Eggplant seedlings transferred outdoors were placed under plastic tarps either to provide relative protection from solar ultraviolet-B (UV-B) radiation (280-315 nm) using Mylar film or to allow exposure to UV-B using cellulose acetate. Protection of seedlings from UV-B radiation resulted in greater leaf expansion than for UV-B-exposed seedlings, but no change in leaf or shoot dry weight occurred after 9 days of treatment. Specific leaf weight increased in response to UV-B exposure outdoors. Exposure of eggplant to UV-B radiation from fluorescent sunlamps in the greenhouse also decreased leaf expansion and leaf and shoot dry weight gain after 5 days of treatment. However, there were no differences in leaf or shoot dry weight relative to control plants after 12 days of UV-B treatment, indicating that UV-B treated plants had acclimated to the treatment and actually had caught up with non-UV-B-irradiated plants in terms of growth.

Localization, trafficking, and temperature-dependence of the Aequorea green fluorescent protein in cultured vertebrate cells.

PubMed Central

Ogawa, H; Inouye, S; Tsuji, F I; Yasuda, K; Umesono, K

1995-01-01

The localization, trafficking, and fluorescence of Aequorea green fluorescent protein (GFP) in cultured vertebrate cells transiently transfected with GFP cDNA were studied. Fluorescence of GFP in UV light was found to be strongest when cells were incubated at 30 degrees C but was barely visible at an incubation temperature of 37 degrees C. COS-1 cells, primary chicken embryonic retina cells, and carp epithelial cells were fluorescently labeled under these conditions. GFP was distributed uniformly throughout the cytoplasm and nucleus independent of cell type examined. When GFP was fused to PML protooncogene product, fluorescence was detected in a unique nuclear organelle pattern indistinguishable from that of PML protein, showing the potential use of GFP as a fluorescent tag. To analyze both function and intracellular trafficking of proteins fused to GFP, a GFP-human glucocorticoid receptor fusion construct was prepared. The GFP-human glucocorticoid receptor efficiently transactivated the mouse mammary tumor virus promoter in response to dexamethasone at 30 degrees C but not at 37 degrees C, indicating that temperature is important, even for function of the GFP fusion protein. The dexamethasone-induced translocation of GFP-human glucocorticoid receptor from cytoplasm to nucleus was complete within 15 min; the translocation could be monitored in a single living cell in real time. Images Fig. 1 Fig. 2 Fig. 3 Fig. 5 PMID:8524871

Cellular imaging using BODIPY-, pyrene- and phthalocyanine-based conjugates.

PubMed

Bizet, Faustine; Ipuy, Martin; Bernhard, Yann; Lioret, Vivian; Winckler, Pascale; Goze, Christine; Perrier-Cornet, Jean-Marie; Decréau, Richard A

2018-01-15

Fluorescent Probes aimed at absorbing in the blue/green region of the spectrum and emitting in the green/red have been synthesized (as the form of dyads-pentads), studied by spectrofluorimetry, and used for cellular imaging. The synthesis of phthalocyanine-pyrene 1 was achieved by cyclotetramerization of pyrenyldicyanobenzene, whereas phthalocyanine-BODIPY 2c was synthesized by Sonogashira coupling between tetraiodophthalocyanine and meso-alkynylBODIPY. The standard four-steps BODIPY synthesis was applied to the BODIPY-pyrene dyad 3 starting from pyrenecarbaldehyde and dimethylpyrrole. 1 H, 13 C, 19 F, 11 BNMR, ICP, MS, and UV/Vis spectroscopic analyses demonstrated that 2c is a mixture of BODIPY-Pc conjugates corresponding to an average ratio of 2.5 BODIPY per Pc unit, where its bis, tris, tetrakis components could not be separated. Fluorescence emission studies (μM concentration in THF) showed that the design of the probes allowed excitation of their antenna (pyrene, BODIPY) in the blue/green region of the spectrum, and subsequent transfer to the acceptor platform (BODIPY, phthalocyanine) followed by its emission in the green/red (with up to 140-350 nm overall Stokes shifts). The fluorescent probes were used for cellular imaging of B16F10 melanoma cells upon solubilization in 1% DMSO containing RPMI or upon encapsulation in liposomes (injection method). Probes were used at 1-10 μM concentrations, cells were fixed with methanol and imaged by biphoton and/or confocal microscopy, showing that probes could achieve the staining of cells membranes and not the nucleus. Copyright © 2017. Published by Elsevier Ltd.

Visible light excitable Zn2+ fluorescent sensor derived from an intramolecular charge transfer fluorophore and its in vitro and in vivo application.

PubMed

Qian, Fang; Zhang, Changli; Zhang, Yumin; He, Weijiang; Gao, Xiang; Hu, Ping; Guo, Zijian

2009-02-04

The UV- and sensor-induced interferences to living systems pose a barrier for in vivo Zn(2+) imaging. In this work, an intramolecular charge transfer (ICT) fluorophore of smaller aromatic plane, 4-amino-7-nitro-2,1,3-benzoxadiazole, was adopted to construct visible light excited fluorescent Zn(2+) sensor, NBD-TPEA. This sensor demonstrates a visible ICT absorption band, a large Stokes shift, and biocompatibility. It emits weakly (Phi = 0.003) without pH dependence at pH 7.1-10.1, and the lambda(ex) and lambda(em) are 469 (epsilon(469) = 2.1 x 10(4) M(-1) cm(-1)) and 550 nm, respectively. The NBD-TPEA displays distinct selective Zn(2+)-amplified fluorescence (Phi = 0.046, epsilon(469) = 1.4 x 10(4) M(-1) cm(-1)) with emission shift from 550 to 534 nm, which can be ascribed to the synergic Zn(2+) coordination by the outer bis(pyridin-2-ylmethyl)amine (BPA) and 4-amine. The Zn(2+) binding ratio of NBD-TPEA is 1:1. By comparison with its analogues NBD-BPA and NBD-PMA, which have no Zn(2+) affinity, the outer BPA in NBD-TPEA should be responsible for the Zn(2+)-induced photoinduced electron transfer blockage as well as for the enhanced Zn(2+) binding ability of 4-amine. Successful intracellular Zn(2+) imaging on living cells with NBD-TPEA staining exhibited a preferential accumulation at lysosome and Golgi with dual excitability at either 458 or 488 nm. The intact in vivo Zn(2+) fluorescence imaging on zebrafish embryo or larva stained with NBD-TPEA revealed two zygomorphic luminescent areas around its ventricle which could be related to the Zn(2+) storage for the zebrafish development. Moreover, high Zn(2+) concentration in the developing neuromasters of zebrafish can be visualized by confocal fluorescence imaging. This study demonstrates a novel strategy to construct visible light excited Zn(2+) fluorescent sensor based on ICT fluorophore other than xanthenone analogues. Current data show that NBD-TPEA staining can be a reliable approach for the intact in vivo Zn(2+) imaging of zebrafish larva as well as for the clarification of subcellular distribution of Zn(2+) in vitro.

Disposal of Energy by UV-B Sunscreens

NASA Astrophysics Data System (ADS)

Nordlund, Thomas; Krishnan, Rajagopal

2008-03-01

Ideal sunscreens absorb dangerous UV light and dispose of the energy safely. ``Safe disposal'' usually means conversion to heat. However, efficient absorption entails a high radiative rate, which implies high energy-transfer and other rates, unless some process intervenes to ``defuse'' the excited state. We studied the excited-state kinetics of three UV-B (290-320 nm) sunscreens by absorption, steady-state and time-resolved fluorescence. Excited-state rate analysis suggests that some sunscreens have low radiative-rate ``dark'' states, in addition to normal excited states.* We deduce dark states when sunscreens of high extinction coefficient do not show lifetimes and total emission consistent with such high radiative rates. A high radiative rate, accompanied by efficient fluorescence emission and/or transfer, may be unfavorable for a sunscreen. In spite of its dark excited state, padimate O shows significant re-emission of light in the UV-A (320-400 nm) and energy transfer to a natural component of excised skin, probably collagen. * Krishnan, R. and T.M. Nordlund (2007) J. Fluoresc. DOI 10.1007/s10895-007-0264-3.

Visualizing different uranium oxidation states during the surface alteration of uraninite and uranium tetrachloride.

PubMed

Grossmann, Kay; Arnold, Thuro; Steudtner, Robin; Weiss, Stefan; Bernhard, Gert

2009-08-01

Low-temperature alteration reactions on uranium phases may lead to the mobilization of uranium and thereby poses a potential threat to humans living close to uranium-contaminated sites. In this study, the surface alteration of uraninite (UO(2)) and uranium tetrachloride (UCl(4)) in air atmosphere was studied by confocal laser scanning microscopy (CLSM) and laser-induced fluorescence spectroscopy using an excitation wavelength of 408 nm. It was found that within minutes the oxidation state on the surface of the uraninite and the uranium tetrachloride changed. During the surface alteration process U(IV) atoms on the uraninite and uranium tetrachloride surface became stepwise oxidized by a one-electron step at first to U(V) and then further to U(VI). These observed changes in the oxidation states of the uraninite surface were microscopically visualized and spectroscopically identified on the basis of their fluorescence emission signal. A fluorescence signal in the wavelength range of 415-475 nm was indicative for metastable uranium(V), and a fluorescence signal in the range of 480-560 nm was identified as uranium(VI). In addition, the oxidation process of tetravalent uranium in aqueous solution at pH 0.3 was visualized by CLSM and U(V) was fluorescence spectroscopically identified. The combination of microscopy and fluorescence spectroscopy provided a very convincing visualization of the brief presence of U(V) as a metastable reaction intermediate and of the simultaneous coexistence of the three states U(IV), U(V), and U(VI). These results have a significant importance for fundamental uranium redox chemistry and should contribute to a better understanding of the geochemical behavior of uranium in nature.

Targeting N-doped graphene quantum dot with high photothermal conversion efficiency for dual-mode imaging and therapy in vitro.

PubMed

Xuan, Yang; Zhang, Ruo-Yun; Zhang, Xiao-Shuai; An, Jie; Cheng, Kai; Li, Cheng; Hou, Xiao-Lin; Zhao, Yuan-Di

2018-08-31

A graphene quantum dot (GQD) is a novel carbon nanomaterial with the advantages of low cost and no pollution. It has attracted serious attention in the biomedical fields because of its stabilities and tunable fluorescence wavelength. In this manuscript, an N-doped graphene quantum dot (N-GQD) was synthesized by a hydrothermal method using citric acid as the carbon source and urea as the nitrogen source. X-ray diffraction, Raman spectroscopy, transmission electron microscopy, UV-vis absorption spectrum, and fluorescence spectrum were used to characterize the N-GQD. The results showed that the N-GQD had a uniform size of about 5 nm. The two fluorescence emission peaks, one in the visible light region showed a 49.75% quantum yield, while another in the near infrared region was 2.49%. The photothermal conversion efficiency was 62.53%, higher than any kind of carbon nanomaterial in existence today. MTT and a long-term cytotoxicity experiment confirmed that the N-GQD had low cytotoxicity. The probe also had the ability of photoacoustic response at the same time. After coupling with folic acid, it presented imaging and photothermal therapy on the cells, which has great application prospects in the early diagnosis and treatment of tumors.

Two photon spectroscopy and microscopy of the fluorescent flavoprotein, iLOV.

PubMed

Homans, Rachael J; Khan, Raja U; Andrews, Michael B; Kjeldsen, Annemette E; Natrajan, Louise S; Marsden, Steven; McKenzie, Edward A; Christie, John M; Jones, Alex R

2018-06-06

LOV-domains are ubiquitous photosensory proteins that are commonly re-engineered to serve as powerful and versatile fluorescent proteins and optogenetic tools. The photoactive, flavin chromophore, however, is excited using short wavelengths of light in the blue and UV regions, which have limited penetration into biological samples and can cause photodamage. Here, we have used non-linear spectroscopy and microscopy of the fluorescent protein, iLOV, to reveal that functional variants of LOV can be activated to great effect by two non-resonant photons of lower energy, near infrared light, not only in solution but also in biological samples. The two photon cross section of iLOV has a significantly blue-shifted S0 → S1 transition compared with the one photon absorption spectrum, suggesting preferential population of excited vibronic states. It is highly likely, therefore, that the two photon absorption wavelength of engineered, LOV-based tools is tuneable. We also demonstrate for the first time two photon imaging using iLOV in human epithelial kidney cells. Consequently, two photon absorption by engineered, flavin-based bio-molecular tools can enable non-invasive activation with high depth resolution and the potential for not only improved image clarity but also enhanced spatiotemporal control for optogenetic applications.

Planetary nebulae with UVIT: Far ultra-violet halo around the Bow Tie nebula (NGC 40)

NASA Astrophysics Data System (ADS)

Kameswara Rao, N.; Sutaria, F.; Murthy, J.; Krishna, S.; Mohan, R.; Ray, A.

2018-01-01

Context. NGC 40 is a planetary nebula with diffuse X-ray emission, suggesting an interaction of the high-speed wind from WC8 central star (CS) with the nebula. It shows strong C IV 1550 Å emission that cannot be explained by thermal processes alone. We present here the first map of this nebula in C IV emission using broad band filters on the Ultra-Violet Imaging Telescope (UVIT). Aim. We aim to map the hot C IV-emitting gas and its correspondence with soft X-ray (0.3-8 keV) emitting regions in order to study the shock interaction between the nebula and the ISM. We also aim to illustrate the potential of UVIT for nebular studies. Methods: We carry out a morphological study of images of the nebula obtained at an angular resolution of about 1.3″ in four UVIT filter bands that include C IV 1550 Å and [C II] 2326 Å lines as well as UV continuum. We also make comparisons with X-ray, optical, and IR images from the literature. Results: The [C II] 2326 Å images show the core of the nebula with two lobes on either side of CS similar to [N II]. The C IV emission in the core shows similar morphology and extent to that of diffuse X-ray emission concentrated in nebular condensations. A surprising UVIT discovery is the presence of a large faint far UV (FUV) halo in an FUV filter with λeff of 1608 Å. The UV halo is not present in any other UV filter. The FUV halo is most likely due to UV fluorescence emission from the Lyman bands of H2 molecules. Unlike the optical and IR halo, the FUV halo trails predominantly towards the south-east side of the nebular core, opposite to the CS's proper motion direction. Conclusions: Morphological similarity of C IV 1550 Å and X-ray emission in the core suggests that it results mostly from the interaction of strong CS wind with the nebula. The FUV halo in NGC 40 highlights the extensive existence of H2 molecules in the regions even beyond the optical and IR halos. Thus UV studies are important to estimate the amount of H2, which is probably the most dominant molecule and significant for mass-loss studies. Based on data obtained with the Ultra-Violet Imaging Telescope (UVIT) on the ASTROSAT satellite.

Fluorescence-Assisted Gamma Spectrometry for Surface Contamination Analysis

NASA Astrophysics Data System (ADS)

Ihantola, Sakari; Sand, Johan; Perajarvi, Kari; Toivonen, Juha; Toivonen, Harri

2013-02-01

A fluorescence-based alpha-gamma coincidence spectrometry approach has been developed for the analysis of alpha-emitting radionuclides. The thermalization of alpha particles in air produces UV light, which in turn can be detected over long distances. The simultaneous detection of UV and gamma photons allows detailed gamma analyses of a single spot of interest even in highly active surroundings. Alpha particles can also be detected indirectly from samples inside sealed plastic bags, which minimizes the risk of cross-contamination. The position-sensitive alpha-UV-gamma coincidence technique reveals the presence of alpha emitters and identifies the nuclides ten times faster than conventional gamma spectrometry.

Robust and economical multi-sample, multi-wavelength UV/vis absorption and fluorescence detector for biological and chemical contamination

NASA Astrophysics Data System (ADS)

Lu, Peter J.; Hoehl, Melanie M.; Macarthur, James B.; Sims, Peter A.; Ma, Hongshen; Slocum, Alexander H.

2012-09-01

We present a portable multi-channel, multi-sample UV/vis absorption and fluorescence detection device, which has no moving parts, can operate wirelessly and on batteries, interfaces with smart mobile phones or tablets, and has the sensitivity of commercial instruments costing an order of magnitude more. We use UV absorption to measure the concentration of ethylene glycol in water solutions at all levels above those deemed unsafe by the United States Food and Drug Administration; in addition we use fluorescence to measure the concentration of d-glucose. Both wavelengths can be used concurrently to increase measurement robustness and increase detection sensitivity. Our small robust economical device can be deployed in the absence of laboratory infrastructure, and therefore may find applications immediately following natural disasters, and in more general deployment for much broader-based testing of food, agricultural and household products to prevent outbreaks of poisoning and disease.

Numerical Study on the Particle Trajectory Tracking in a Micro-UV Bio-Fluorescence Sensor.

PubMed

Byeon, Sun-Seok; Cho, Moon-Young; Lee, Jong-Chul; Kim, Youn-Jea

2015-03-01

A micro-UV bio-fluorescence sensor was developed to detect primary biological aerosols including bacteria, bacterial spores, fungal spores, pollens, viruses, algae, etc. In order to effectively detect the bio-particles in a micro-UV bio-fluorescence sensor, numerical calculations were performed to adjust for appropriate flow conditions of the sensor by regulating the sample aerosols and sheath flow. In particular, a CFD-based model of hydrodynamic processes was developed by computing the trajectory of particles using commercially available ANSYS CFX-14 software and the Lagrangian tracking model. The established model was evaluated with regard to the variation of sheath flow rate and particle size. Results showed that the sheath flow was changed rapidly at the end of nozzle tip, but the sample particles moved near the center of aerosol jet for aerodynamic focusing with little deviation from the axis.

Effects of TiO2 nanoparticles on ROS production and growth inhibition using freshwater green algae pre-exposed to UV irradiation.

PubMed

Fu, Ling; Hamzeh, Mahsa; Dodard, Sabine; Zhao, Yuan H; Sunahara, Geoffrey I

2015-05-01

This study investigated the possibility that titanium dioxide nanoparticles (nano-TiO2) toxicity in Pseudokirchneriella subcapitata involves reactive oxygen species (ROS) production, using the dichlorodihydrofluorescein (DCF) assay. Algae were exposed to nano-TiO2 under laboratory fluorescent lamps supplemented with UV irradiation for 3h, with or without a UV filter. Results showed that nano-TiO2 increased ROS production in UV-exposed cells, with or without a UV filter (LOEC values were 250 and 10mg/L, respectively). Sublethal effects of nano-TiO2 on UV pre-exposed algae were also examined. Toxicity studies indicated that exposure to nano-TiO2 agglomerates decreased algal growth following 3h pre-exposure to UV, with or without a UV filter (EC50s were 8.7 and 6.3mg/L, respectively). The present study suggests that the growth inhibitory effects of nano-TiO2 in algae occurred at concentrations lower than those that can elevate DCF fluorescence, and that ROS generation is not directly involved with the sublethal effects of nano-TiO2 in algae. Copyright © 2015 Elsevier B.V. All rights reserved.

Spectral studies on the interaction between lanthanum ion and the ligand: N,N'-ethylenebis-[2-(o-hydroxyphenolic)glycine].

PubMed

Yaqin, Zhao; Binsheng, Yang

2005-11-01

The interaction between N,N'-ethylenebis-[2-(o-hydroxyphenolic)glycine] (EHPG) and lanthanum was studied by the difference UV spectra and fluorescence spectra. At pH 7.4, 0.01 M N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (Hepes), with the addition of 1.0 x 10(-3)M lanthanum, two new peaks were observed at 238 nm and 294 nm by absorptivity spectroscopy compared with blank solution EHPG suggesting the interaction of lanthanum and EHPG. At the same time, the reaction could be measured by fluorescence spectra. The fluorescence intensity of EHPG at 310 nm was significantly decreased in the presence of lanthanum. The 1:1 stoichiometric ratio of EHPG to lanthanum was confirmed by both fluorescence and UV titration curves. In addition, the molar absorptivity of La-EHPG at 238 nm is (1.23+/-0.01)x10(4)cm(-1)M(-1). The conditional binding constant was calculated to be log K(La-EHPG)=12.09+/-0.37 on the basis of the result of UV titration curves.

A Real-Time Ultraviolet Radiation Imaging System Using an Organic Photoconductive Image Sensor†

PubMed Central

Okino, Toru; Yamahira, Seiji; Yamada, Shota; Hirose, Yutaka; Odagawa, Akihiro; Kato, Yoshihisa; Tanaka, Tsuyoshi

2018-01-01

We have developed a real time ultraviolet (UV) imaging system that can visualize both invisible UV light and a visible (VIS) background scene in an outdoor environment. As a UV/VIS image sensor, an organic photoconductive film (OPF) imager is employed. The OPF has an intrinsically higher sensitivity in the UV wavelength region than those of conventional consumer Complementary Metal Oxide Semiconductor (CMOS) image sensors (CIS) or Charge Coupled Devices (CCD). As particular examples, imaging of hydrogen flame and of corona discharge is demonstrated. UV images overlapped on background scenes are simply made by on-board background subtraction. The system is capable of imaging weaker UV signals by four orders of magnitude than that of VIS background. It is applicable not only to future hydrogen supply stations but also to other UV/VIS monitor systems requiring UV sensitivity under strong visible radiation environment such as power supply substations. PMID:29361742

Composite fluorescent nanoparticles for biomedical imaging.

PubMed

Pansare, Vikram J; Bruzek, Matthew J; Adamson, Douglas H; Anthony, John; Prud'homme, Robert K

2014-04-01

In the rapidly expanding field of biomedical imaging, there is a need for nontoxic, photostable, and nonquenching fluorophores for fluorescent imaging. We have successfully encapsulated a new, extremely hydrophobic, pentacene-based fluorescent dye within polymeric nanoparticles (NPs) or nanocarriers (NCs) via the Flash NanoPrecipitation (FNP) process. Nanoparticles and dye-loaded micelles were formulated by FNP and characterized by dynamic light scattering, fluorescence spectroscopy, UV-VIS absorbance spectroscopy, and confocal microscopy. These fluorescent particles were loaded from less than 1% to 78% by weight core loading and the fluorescence maximum was found to be at 2.3 wt.%. The particles were also stably formed at 2.3% core loading from 20 up to 250 nm in diameter with per-particle fluorescence scaling linearly with the NC core volume. The major absorption peaks are at 458, 575, and 625 nm, and the major emission peaks at 635 and 695 nm. In solution, the Et-TP5 dye displays a strong concentration-dependent ratio of the emission intensities of the first two emission peaks, whereas in the nanoparticle core the spectrum is independent of concentration over the entire concentration range. A model of the fluorescence quenching was consistent with Förster resonant energy transfer as the cause of the quenching observed for Et-TP5. The Förster radius calculated from the absorption and emission spectra of Et-TP5 is 4.1 nm, whereas the average dye spacing in the particles at the maximum fluorescence is 3.9 nm. We have successfully encapsulated Et-TP5, a pentacene derivative dye previously only used in light-emitting diode applications, within NCs via the FNP process. The extreme hydrophobicity of the dye keeps it encapsulated in the NC core, its extended pentacene structure gives it relatively long wavelength emission at 695 nm, and the pentacene structure, without oxygen or nitrogen atoms in its core, makes it highly resistant to photobleaching. Its bulky side groups minimize self-quenching and localization within the nanoparticle core prevents interaction of the dye with biological surfaces, or molecules in diagnostic assays. Loading of dye in the NP core allows 25 times more dye to be delivered than if it were conjugated onto the nanocarrier surface. The utility of the dye for quantifying nanoparticle binding is demonstrated. Studies to extend the wavelength range of these pentacene dyes into the near infra-red are underway.

UV laser-induced fluorescence spectroscopy and laser Doppler flowmetry in the diagnostics of alopecia

NASA Astrophysics Data System (ADS)

Skomorokha, Diana P.; Pigoreva, Yulia N.; Salmin, Vladimir V.

2016-04-01

Development of optical biopsy methods has a great interest for medical diagnostics. In clinical and experimental studies it is very important to analyze blood circulation quickly and accurately, thereby laser Doppler flowmetry (LDF) is widely used. UV laser-induced fluorescence spectroscopy (UV LIFS) is express highly sensitive and widely-spread method with no destructive impact, high excitation selectivity and the possibility to use in highly scattering media. The goal of this work was to assess a correlation of UV laser-induced fluorescence spectroscopy and laser Doppler flowmetry parameters, and a possibility to identify or to differentiate various types of pathological changes in tissues according to their autofluorescence spectra. Three groups of patients with diffuse (symptomatic) alopecia, androgenic alopecia, and focal alopecia have been tested. Each groups consisted of not less than 20 persons. The measurements have been done in the parietal and occipital regions of the sculls. We used the original automated spectrofluorimeter to record autofluorescence spectra, and standard laser Doppler flowmeter BLF-21 (Transonic Systems, Inc., USA) to analyze the basal levels of blood circulation. Our results show that UV LIFS accurately distinguishes the zones with different types of alopecia. We found high correlation of the basal levels of blood circulation and the integrated intensity of autofluorescence in the affected tissue.

Synthesis, characterization, and spectroscopic investigation of benzoxazole conjugated Schiff bases.

PubMed

Santos, Fabiano S; Costa, Tania M H; Stefani, Valter; Gonçalves, Paulo F B; Descalzo, Rodrigo R; Benvenutti, Edilson V; Rodembusch, Fabiano S

2011-11-24

Two Schiff bases were synthesized by reaction of 2-(4'-aminophenyl)benzoxazole derivatives with 4-N,N-diethylaminobenzaldehyde. UV-visible (UV-vis) and steady-state fluorescence in solution were applied in order to characterize its photophysical behavior. The Schiff bases present absorption in the UV region with fluorescence emission in the blue-green region, with a large Stokes' shift. The UV-vis data indicates that each dye behaves as two different chromophores in solution in the ground state. The fluorescence emission spectra of the dye 5a show that an intramolecular proton transfer (ESIPT) mechanism takes place in the excited state, whereas a twisted internal charge transfer (TICT) state is observed for the dye 5b. Theoretical calculations were performed in order to study the conformation and polarity of the molecules at their ground and excited electronic states. Using density functional theory (DFT) methods at theoretical levels BLYP/Aug-SV(P) for geometry optimizations and B3LYP/6-311++G(2d,p) for single-point energy evaluations, the calculations indicate that the lowest energy conformations are in all cases nonplanar and that the dipole moments of the excited state relaxed structures are much larger than those of the ground state structures, which corroborates the experimental UV-vis absorption results.

Autofluorescence-Free Live-Cell Imaging Using Terbium Nanoparticles.

PubMed

Cardoso Dos Santos, M; Goetz, J; Bartenlian, H; Wong, K-L; Charbonnière, L J; Hildebrandt, N

2018-04-18

Fluorescent nanoparticles (NPs) have become irreplaceable tools for advanced cellular and subcellular imaging. While very bright NPs require excitation with UV or visible light, which can create strong autofluorescence of biological components, NIR-excitable NPs without autofluorescence issues exhibit much lower brightness. Here, we show the application of a new type of surface-photosensitized terbium NPs (Tb-NPs) for autofluorescence-free intracellular imaging in live HeLa cells. The combination of exceptionally high brightness, high photostability, and long photoluminecence (PL) lifetimes for highly efficient suppression of the short-lived autofluorescence allowed for time-gated PL imaging of intracellular vesicles over 72 h without toxicity and at extremely low Tb-NP concentrations down to 12 pM. Detection of highly resolved long-lifetime (ms) PL decay curves from small (∼10 μm 2 ) areas within single cells within a few seconds emphasized the unprecedented photophysical properties of Tb-NPs for live-cell imaging that extend well beyond currently available nanometric imaging agents.

The Chemistry and Perception of Fluorescent White Textile Materials

NASA Astrophysics Data System (ADS)

Xu, Changhai

Cationic bleach activators (CBA) are precursors to perhydroxyl compounds that, when activated, have higher oxidation potential and potentially improved bleaching performance compared to common oxidizing agents such as hydrogen peroxide. CBAs were first reported in 1997 by Proctor & Gamble Co., and have been further developed at North Carolina State University. To date, an effective low temperature bleaching system has not been developed that offers sufficient economic improvement over existing bleaching systems. The primary purpose of this research was to develop new methods and understand key variables required for achieving enhanced whiteness of textile materials using bleach activators with or without the presence of fluorescence. A new optimized low temperature bleaching system using novel lactam-based bleach activators was developed and the effect of UV content of light sources on the whiteness of fluorescent white textile materials was evaluated. A novel class of bleach activators was synthesized by introducing benzoylcaprolactam group into a stilbene system shown as follows:* While solubility, purification and hydrolytic stability of the compounds were problematic, a new approach to low pH (pH 7-9) and low temperature (50-70°C) bleaching was found using a butyrolactam-based cationic bleach activator, N-[4-(triethylammonio methyl)benzoyl]-butyrolactam chloride (TBBC), using a central composite design (CCD) of experiment. The CCD bleaching experiments showed that cationic bleach activators are less effective with high concentrations of hydrogen peroxide in high alkaline solutions. Also a 2FI model predicted the optimized bleaching performance on 100% cotton at near neutral pH and temperatures around 50°C, in which the dosage of TBBC is the most important factor affecting the bleaching performance. This prediction was validated experimentally during bleaching of bamboo and cotton fibers. In addition, this study confirmed the hypothesis that cationic bleach activators have inherent substantivity to cellulosic fibers and that the substantivity enhances bleach effectiveness. This cationic activated bleach system was effective for cotton bleaching and the bleaching performance is superior to that of conventional peroxide bleaching. TBBC was also applied to bamboo cellulosic fibers, which exhibit excessive strength loss during conventional hot bleaching. Under optimized conditions of 5 g/L TBBC, 50°C, 30 min and pH 7.0, TBBC-based bleaching of bamboo fibers produced CIE Whiteness Index (CIE WI) values of 58.20 compared to untreated bamboo which had CIE WI values of 10.77. Hence, the TBBC bleach activator method is effective at bleaching bamboo fibers. Besides chemical bleaching, the fluorescent whitening was another approach to increase the whiteness of materials. Since the whiteness of fluorescent white materials is produced by absorbing UV light and emitting visible blue light, the UV content of light sources has a significant effect on the perception of whiteness. This research addresses the common light sources used in color matching booths. The pilot data is collected by measuring spectral radiance and spectral irradiance, which is used for analysis of the UV effect on the whiteness of fluorescent white materials. The whitening performance of a fluorescent brightening agent (FBA) is dependent on the energy and intensity of the incident UV light. No data have been reported in the open literature that shows the UV emission of standard lamps used in viewing booths. Indeed, standards pertaining to lighting do not require or recommend the standardization of the UV content in any lamps. Hence, the spectroradiometric quantification of UV emission of a series of standard viewing booths is a requirement for establishment of a methodology to determine the effect of radiometric variability in standard sources on visual perception of fluorescent white materials. The radiance measurement data collected from measuring the radiance of light sources (including daylight simulation, incandescent, horizon daylight, cool white fluorescence, and Ultralume 30) in a SpectraLight III color viewing booth and the irradiance of these light sources over a PTFE diffuse reflectance standard, AATCC textile UV calibration standard and some fluorescent whitened fabric samples showed the inadequacy of UV content of these light sources in the SpectraLight III. *Please refer to dissertation for diagram.

Hg speciation by differential photochemical vapor generation at UV-B and UV-C wavelengths

USDA-ARS?s Scientific Manuscript database

Mercury speciation was accomplished by differential photochemical reduction at two UV wavelengths; the resulting Hg(O) vapor was quantified by atomic fluorescence spectrometry. After microwave digestion and centrifugation, analyte solutions were mixed with 20% (v/v) formic acid in a reactor coil, an...

Scanning lidar fluorosensor for remote diagnostic of surfaces

NASA Astrophysics Data System (ADS)

Caneve, Luisa; Colao, Francesco; Fantoni, Roberta; Fiorani, Luca

2013-08-01

Scanning hyperspectral systems based on laser induced fluorescence (LIF) have been developed and realized at the ENEA allowing to obtain information of analytical and qualitative interest on different materials by the study of the emission of fluorescence. This technique, for a surface analysis, is fast, remote, not invasive and specific. A new compact setup capable of fast 2D monochromatic images acquisition on up to 90 different spectral channels in the visible/UV range will be presented. It has been recently built with the aim to increase the performances in terms of space resolution, time resolved capabilities and data acquisition speed. Major achievements have been reached by a critical review of the optical design. The results recently obtained with in-situ measurements of interest for applications in the field of cultural heritage will be shown. 2001 Elsevier Science. All rights reserved

Binding Mode and Selectivity of a Scorpiand-Like Polyamine Ligand to Single- and Double-Stranded DNA and RNA: Metal- and pH-Driven Modulation.

PubMed

Inclán, Mario; Guijarro, Lluis; Pont, Isabel; Frías, Juan C; Rotger, Carmen; Orvay, Francisca; Costa, Antoni; García-España, Enrique; Albelda, M Teresa

2017-11-13

The interaction of a polyazacyclophane ligand having an ethylamine pendant arm functionalized with an anthryl group (L), with the single-stranded polynucleotides polyA, polyG, polyU, and polyC as well as with the double-stranded polynucleotides polyA-polyU, poly(dAT) 2 , and poly(dGC) 2 has been followed by UV/Vis titration, steady state fluorescence spectroscopy, and thermal denaturation measurements. In the case of the single-stranded polynucleotides, the UV/Vis and fluorescence titrations permit to distinguish between sequences containing purine and pyrimidine bases. For the double-stranded polynucleotides the UV/Vis measurements show for all of them hypochromicity and bathochromic shifts. However, the fluorescence studies reveal that both polyA-polyU and poly(dAT) 2 induce a twofold increase in the fluorescence, whereas interaction of poly(dGC) 2 with the ligand L induces a quenching of the fluorescence. Cu 2+ modulates the interaction with the double-stranded polynucleotides due to the conformation changes that its coordination induces in compound L. In general, the spectroscopic studies show that intercalation seems to be blocked by the formation of the metal complex. All these features suggest the possibility of using compound L as a sequence-selective fluorescence probe. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Spiropyran-Isoquinoline Dyad as a Dual Chemosensor for Co(II) and In(III) Detection.

PubMed

Kho, Yong-Min; Shin, Eun Ju

2017-09-19

Spiropyran derivatives have been studied as light-regulated chemosensors for a variety of metal cations and anions, but there is little research on chemosensors that simultaneously detect multiple metal cations. In this study, a spiropyran derivative with isoquinoline, SP-IQ , was prepared and it functions investigated as a light-regulated sensor for both Co 2+ and In 3+ cations. A colorless nonfluorescent SP-IQ converts to a pink-colored fluorescent MC-IQ by UV irradiation or standing in the dark, and MC-IQ returns to SP-IQ with visible light. Upon UV irradiation with the Co 2+ cation for 7 min, the stronger absorption at 540 nm and the similar fluorescence intensity at 640 nm are observed, compared to when no metal cation is added, due to the formation of a Co 2+ complex with pink color and pink fluorescence. When placed in the dark with the In 3+ cation for 7 h, the colorless solution of SP-IQ changes to the In 3+ complex with yellow color and pink fluorescence, which shows strong absorption at 410 nm and strong fluorescence at 640 nm. Selective detection of the Co 2+ cation with UV irradiation and the In 3+ cation in the dark could be possible with SP-IQ by both absorption and fluorescence spectroscopy or by the naked eye.

Image-guided intervention in the human bile duct using scanning fiber endoscope system

NASA Astrophysics Data System (ADS)

Seibel, Eric J.; Jo, Javier A.; Melville, C. David; Johnston, Richard S.; Naumann, Christopher R.; Saunders, Michael D.

2012-01-01

Bile duct cancers are increasing in frequency while being difficult to diagnose. Currently available endoscopic imaging devices used in the biliary tree are low resolution with poor image quality, leading to inadequate evaluation of indeterminate biliary strictures. However, a new ultrathin and flexible cholangioscope system has been successfully demonstrated in a human subject. This mini-cholangioscope system uses a scanning fiber endoscope (SFE) as a forward-imaging guidewire, dimensions of 1.2-mm diameter and 3-m length. Full color video (500-line resolution at 30Hz) is the standard SFE imaging mode using spiral scanning of red, green, and blue laser light at low power. Image-guided operation of the biopsy forceps was demonstrated in healthy human bile ducts with and without saline flushing. The laser-based video imaging can be switched to various modes to enhance tissue markers of disease, such as widefield fluorescence and enhanced spectral imaging. In parallel work, biochemical discrimination of tissue health in pig bile duct has been accomplished using fiberoptic delivery of pulsed UV illumination and time-resolved autofluorescence spectroscopic measurements. Implementation of time-resolved fluorescence spectroscopy for biochemical assessment of the bile duct wall is being done through a secondary endoscopic channel. Preliminary results indicate that adequate SNR levels (> 30 dB) can be achieved through a 50 micron fiber, which could serve as an optical biopsy probe. The SFE is an ideal mini-cholangioscope for integration of both tissue and molecular specific image contrast in the future. This will provide the physician with unprecedented abilities to target biopsy locations and perform endoscopically-guided therapies.

The imprints of the high light and UV-B stresses in Oryza sativa L. 'Kanchana' seedlings are differentially modulated.

PubMed

Faseela, Parammal; Puthur, Jos T

2018-01-01

High light and ultraviolet-B radiation (UV-B) are generally considered to have negative impact on photosynthesis and plant growth. The present study evaluates the tolerance potential of three cultivars of Oryza sativa L. (Kanchana, Mattatriveni and Harsha) seedlings towards high light and UV-B stress on the basis of photosynthetic pigment degradation, chlorophyll a fluorescence parameters and rate of lipid peroxidation, expressed by malondialdehyde content. Surprisingly, it was revealed that Kanchana was the most sensitive cultivar towards high light and at the same time it was the most tolerant cultivar towards UV-B stress. This contrasting feature of Kanchana towards high light and UV-B tolerance was further studied by analyzing photosystem (PS) I and II activity, mitochondrial activity, chlorophyll a fluorescence transient, enzymatic and non-enzymatic antioxidant defense system. Due to the occurrence of more PS I and PSII damages, the inhibition of photochemical efficiency and emission of dissipated energy as heat or fluorescence per PSII reaction center was higher upon high light exposure than UV-B treatments in rice seedlings of Kanchana. The mitochondrial activity was also found to be drastically altered upon high light as compared to UV-B treatments. The UV-B induced accumulation of non-enzymatic antioxidants (proline, total phenolics, sugar and ascorbate) and enzymatic antioxidants (ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase and glutathione reductase) in rice seedlings than those subjected to high light exposure afforded more efficient protection against UV-B radiation in rice seedlings. Our results proved that high tolerance of Kanchana towards UV-B than high light treatments, correlated linearly with the protected photosynthetic and mitochondrial machinery which was provided by upregulation of antioxidants particularly by total phenolics, ascorbate and ascorbate peroxidase in rice seedlings. Data presented in this study conclusively proved that rice cultivar Kanchana respond to different environmental signals independently and tolerance mechanisms to individual stress factors was also varied. Copyright © 2017 Elsevier B.V. All rights reserved.

Resolution Limits of Nanoimprinted Patterns by Fluorescence Microscopy

NASA Astrophysics Data System (ADS)

Kubo, Shoichi; Tomioka, Tatsuya; Nakagawa, Masaru

2013-06-01

The authors investigated optical resolution limits to identify minimum distances between convex lines of fluorescent dye-doped nanoimprinted resist patterns by fluorescence microscopy. Fluorescent ultraviolet (UV)-curable resin and thermoplastic resin films were transformed into line-and-space patterns by UV nanoimprinting and thermal nanoimprinting, respectively. Fluorescence immersion observation needed an immersion medium immiscible to the resist films, and an ionic liquid of triisobutyl methylphosphonium tosylate was appropriate for soluble thermoplastic polystyrene patterns. Observation with various numerical aperture (NA) values and two detection wavelength ranges showed that the resolution limits were smaller than the values estimated by the Sparrow criterion. The space width to identify line patterns became narrower as the line width increased. The space width of 100 nm was demonstrated to be sufficient to resolve 300-nm-wide lines in the detection wavelength range of 575-625 nm using an objective lens of NA= 1.40.

UV induced visual cues in grasses

PubMed Central

Baby, Sabulal; Johnson, Anil John; Govindan, Balaji; Lukose, Sujith; Gopakumar, Bhaskaran; Koshy, Konnath Chacko

2013-01-01

Grasses are traditionally considered as wind pollinated, however, field observations confirmed frequent insect visits to grass flowers, suggesting insect pollination. Fruit and seed predators inflict heavy losses to cereals and millets during their growth, maturation and storage. The actual factors guiding insects and predators to grass flowers, fruits and seeds are not clear. Here, we report attractive blue fluorescence emissions on grass floral parts such as glumes, lemma, palea, lodicules, staminal filaments, pollens and fruits in ultraviolet (UV) 366 nm, whereas the stigmatic portions were not blue, but red fluorescent. We characterized the blue fluorescent constituent in grass reproductive structures as ferulic acid (FA). Fluorescence spectra of blue-emitting grass floral, seed extracts and isolated FA on excitation at 366 nm showed their emissions at 420–460 nm. We propose these FA-based blue fluorescence emissions in grass reproductive structures as visual cues that attract pollinators, predators and even pests towards them. PMID:24061408

Enhanced broadband near-infrared luminescence from Pr3+-doped tellurite glass with silver nanoparticles

NASA Astrophysics Data System (ADS)

Cheng, Pan; Zhou, Yaxun; Zhou, Minghan; Su, Xiue; Zhou, Zizhong; Yang, Gaobo

2017-11-01

Pr3+-doped tellurite glasses containing metallic silver NPs were synthesized by the conventional melt-quenching technique. Structural, thermal and optical properties of the synthesized glass samples were characterized by X-Ray diffraction (XRD) curves, Raman spectra, differential scanning calorimeter (DSC) curves, transmission electron microscopy (TEM) images, UV/Vis/NIR absorption and near-infrared fluorescence emission spectra. The XRD curves confirmed the amorphous structural nature of the synthesized glasses, the Raman spectra identified the presence of different vibrational groups, the DSC curves verified the good thermal stability, and the TEM images revealed the nucleated silver NPs with average diameter about 10 nm dispersed in the glass matrix and its surface Plasmon resonance (SPR) absorption band was located at around 510 nm. Besides, Judd-Ofelt intensity parameters Ωt (t = 2, 4, 6) and other important spectroscopic parameters like transition probability, radiative lifetime, branching ratio were calculated to evaluate the radiative properties of Pr3+ levels from the measured optical absorption spectra. It was found that Pr3+-doped tellurite glasses could emit an ultra-broadband fluorescence extending from 1250 to 1650 nm under the 488 nm excitation, and this fluorescence emission increased further with the introduction of silver NPs. The enhanced fluorescence was mainly attributed to the increased local electric field around Pr3+ induced by silver NPs. The present results demonstrate that Pr3+-Ag codoped tellurite glass is a promising candidate for the near-infrared band ultra-broadband fiber amplifiers covering the expanded low-loss communication window.

Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

NASA Astrophysics Data System (ADS)

Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

2014-08-01

Primary biological aerosol particles (PBAPs) can contribute significantly to the coarse particle burden in many environments. PBAPs can thus influence climate and precipitation systems as cloud nuclei and can spread disease to humans, animals, and plants. Measurement data and techniques for PBAPs in natural environments at high time- and size resolution are, however, sparse, and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in southwestern Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of a waveband integrated bioaerosol sensor (WIBS-4) with a ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behavior, with increased fluorescent bioparticle concentrations at night, when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each was correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multimodal distributions turning into a broad featureless single mode after averaging, and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent particles. Cladosporium spp., which are among the most abundant fungal spores in many terrestrial environments, were not correlated with any of the real-time fluorescence channels, suggesting that the real-time fluorescence instruments are relatively insensitive to PBAP classes with dark, highly absorptive cell walls. Fluorescence microscopy images of cascade impactor plates showed large numbers of coarse-mode particles consistent with the morphology and weak fluorescence expected of sea salt. Some of these particles were attached to biological cells, suggesting that a marine source influenced the PBAPs observed at the site and that the ocean may be an important contributor to PBAP loadings in coastal environments.

Ambient measurements of biological aerosol particles near Killarney, Ireland: a comparison between real-time fluorescence and microscopy techniques

NASA Astrophysics Data System (ADS)

Healy, D. A.; Huffman, J. A.; O'Connor, D. J.; Pöhlker, C.; Pöschl, U.; Sodeau, J. R.

2014-02-01

Primary biological aerosol particles (PBAP) can contribute significantly to the coarse particle burden in many environments, may thus influence climate and precipitation systems as cloud nuclei, and can spread disease to humans, animals, and plants. Measurements of PBAP in natural environments taken at high time- and size- resolution are, however, sparse and so large uncertainties remain in the role that biological particles play in the Earth system. In this study two commercial real-time fluorescence particle sensors and a Sporewatch single-stage particle impactor were operated continuously from 2 August to 2 September 2010 at a rural sampling location in Killarney National Park in south western Ireland. A cascade impactor was operated periodically to collect size-resolved particles during exemplary periods. Here we report the first ambient comparison of the waveband integrated bioaerosol sensor (WIBS-4) with the ultraviolet aerodynamic particle sizer (UV-APS) and also compare these real-time fluorescence techniques with results of fluorescence and optical microscopy of impacted samples. Both real-time instruments showed qualitatively similar behaviour, with increased fluorescent bioparticle concentrations at night when relative humidity was highest and temperature was lowest. The fluorescent particle number from the FL3 channel of the WIBS-4 and from the UV-APS were strongly correlated and dominated by a 3 μm mode in the particle size distribution. The WIBS FL2 channel exhibited particle modes at approx. 1 and 3 μm, and each were correlated with the concentration of fungal spores commonly observed in air samples collected at the site (ascospores, basidiospores, Ganoderma spp.). The WIBS FL1 channel exhibited variable multi-modal distributions turning into a broad featureless single mode after averaging and exhibited poor correlation with fungal spore concentrations, which may be due to the detection of bacterial and non-biological fluorescent particles. Cladosporium spp., which are among the most abundant fungal spores in many terrestrial environments, were not correlated with any of the real-time fluorescence channels, suggesting that the real-time fluorescence instruments are insensitive to PBAP classes with dark, highly absorptive cell walls. Fluorescence microscopy images of cascade impactor plates showed large numbers of coarse mode particles consistent with the morphology and weak fluorescence expected of sea salt. Some of these particles were attached to biological cells, suggesting that a marine source influenced the PBAP observed at the site and that the ocean may be an important contributor to PBAP loadings in coastal environments.

Photophysical studies on curcumin-sophorolipid nanostructures: applications in quorum quenching and imaging

PubMed Central

2018-01-01

Sophorolipid biosurfactants are biodegradable, less toxic and FDA approved. The purified acidic form of sophorolipid is stimuli-responsive with self-assembling properties and used for solubilizing hydrophobic drugs. This study encapsulated curcumin (CU) with acidic sophorolipid (ASL) micelles and analysed using photophysical studies like UV-visible spectroscopy, photoluminescence (PL) spectroscopy and time-correlated single photon counting (TCSPC). TEM images have revealed ellipsoid micelles of approximately 100 nm size and were confirmed by dynamic light scattering. The bacterial fluorescence uptake studies showed the uptake of formed CUASL nanostructures into both Gram-positive and Gram-negative bacteria. They also showed quorum quenching activity against Pseudomonas aeruginosa. The results have demonstrated this system has potential theranostic applications. PMID:29515826

Modification of fluorescence and optical properties of Rhodamine B dye doped PVA/Chitosan polymer blend films

NASA Astrophysics Data System (ADS)

Padmakumari, R.; Ravindrachary, V.; Mahantesha, B. K.; Sagar, Rohan N.; Sahanakumari, R.; Bhajantri, R. F.

2018-05-01

Pure and Rhodamine B doped Poly (vinyl alcohol)/Chitosan composite films are prepared using solution casting method. Fourier transforms infrared spectra (FTIR), Ultraviolet-Visible (UV-Vis), fluorescence studies were used to characterize the prepared polymer films. The FT-IR results show that the appearance of new peaks along with shift in peak positions indicates the interaction of Rhodamine B with PVA-CS blend. Optical absorption edge, band gap and activation energy were determined from UV-Visible studies. The optical absorption edge increases, band gap decreases and activation energy increases with dopant concentration respectively. The corresponding emission spectra were studied using fluorescence spectroscopy. From the fluorescence study the quenching phenomena are observed in emission wavelength range of 607nm-613nm upon excitation with absorption maxima 443nm.

Particle-fluorescence spectrometer for real-time single-particle measurements of atmospheric organic carbon and biological aerosol.

PubMed

Pan, Yong-Le; Pinnick, Ronald G; Hill, Steven C; Chang, Richard K

2009-01-15

A particle-fluorescence spectrometer (PFS) for real-time measurements of single-particle UV-laser-induced fluorescence (UV-LIF) excited with a pulsed (263-nm) laser is reported. The dispersed UV-LIF spectra are measured by a 32-anode PMT detector with spectral coverage from 280-600 nm. The PFS represents a significant improvement over our previous apparatus [Pinnick et al., Atmos. Environ. 2004, 38, 1657] and can (1) measure fluorescence spectra of bacterial particles having light-scattering sizes as small as 1 microm (previously limited to about 3 microm) and so can measure particles with size in the range of 1-10 microm, (2) measure each particle's elastic scattering which can be used to estimate particle size (not available previously), (3) measure single-particle fluorescence spectra with a laser and detector that can record spectra as fast as 90,000/s, although the highest rates we have found experimentally in atmospheric measurements is only several hundred per second (previously limited by detectors to only 25/s), and (4) provide a time stamp for a data block of spectra with time resolution from 10 ms to 10 min. In addition, the PFS has been modified to be more robust, transportable, and smaller. The use of an aerodynamic-focusing sheath inlet nozzle assembly has improved the sample rate. The PFS has been employed to measure UV-LIF spectra from individual atmospheric particles during October-December 2006 and January-May 2008 in New Haven, CT, and during January-May 2007 in Las Cruces, NM.

Fluorescence spectroscopy of dental calculus

NASA Astrophysics Data System (ADS)

Bakhmutov, D.; Gonchukov, S.; Sukhinina, A.

2010-05-01

The aim of the present study was to investigate the fluorescence properties of dental calculus in comparison with the properties of adjacent unaffected tooth structure using both lasers and LEDs in the UV-visible range for fluorescence excitation. The influence of calculus color on the informative signal is demonstrated. The optimal spectral bands of excitation and registration of the fluorescence are determined.

Transfer of ultraviolet photon energy into fluorescent light in the visible path represents a new and efficient protection mechanism of sunscreens

NASA Astrophysics Data System (ADS)

Vergou, Theognosia; Patzelt, Alexa; Richter, Heike; Schanzer, Sabine; Zastrow, Leonhard; Golz, Karin; Doucet, Olivier; Antoniou, Christina; Sterry, Wolfram; Lademann, Juergen

2011-10-01

The development of sunscreens with high sun protection factor (SPF) values but low filter concentrations is the ultimate goal. The purpose of the present study was to investigate why a sunscreen spray and cream with different concentrations of the same UV-filters provided the same SPF. Therefore, the homogeneity of the distribution of both sunscreens was investigated by laser scanning microscopy (LSM) and tape stripping (TS). Additionally, the energy transfer mechanisms of the sunscreens on the skin were analyzed. The TS and LSM showed a better homogeneity of the distribution of the spray. With Wood's light, a total absorption of the irradiation was detected in the spray area. In contrast, after cream treatment, an intensive fluorescent signal was observed. It was demonstrated that this fluorescent signal was caused by nonthermal energy transferred from the UV-filters to one compound of the cream releasing its excitation energy by fluorescence. This nonthermal energy transfer seemed to be the reason for the high efficiency of the cream, which is subjected to thermal relaxation. The transfer of UV photon energy into fluorescent light represents a new approach to increase the efficiency of sunscreens and could form the basis for a new generation of sunscreens.

Comparative evaluation of differential laser-induced perturbation spectroscopy as a technique to discriminate emerging skin pathology

NASA Astrophysics Data System (ADS)

Kozikowski, Raymond T.; Smith, Sarah E.; Lee, Jennifer A.; Castleman, William L.; Sorg, Brian S.; Hahn, David W.

2012-06-01

Fluorescence spectroscopy has been widely investigated as a technique for identifying pathological tissue; however, unrelated subject-to-subject variations in spectra complicate data analysis and interpretation. We describe and evaluate a new biosensing technique, differential laser-induced perturbation spectroscopy (DLIPS), based on deep ultraviolet (UV) photochemical perturbation in combination with difference spectroscopy. This technique combines sequential fluorescence probing (pre- and post-perturbation) with sub-ablative UV perturbation and difference spectroscopy to provide a new spectral dimension, facilitating two improvements over fluorescence spectroscopy. First, the differential technique eliminates significant variations in absolute fluorescence response within subject populations. Second, UV perturbations alter the extracellular matrix (ECM), directly coupling the DLIPS response to the biological structure. Improved biosensing with DLIPS is demonstrated in vivo in a murine model of chemically induced skin lesion development. Component loading analysis of the data indicates that the DLIPS technique couples to structural proteins in the ECM. Analysis of variance shows that DLIPS has a significant response to emerging pathology as opposed to other population differences. An optimal likelihood ratio classifier for the DLIPS dataset shows that this technique holds promise for improved diagnosis of epithelial pathology. Results further indicate that DLIPS may improve diagnosis of tissue by augmenting fluorescence spectra (i.e. orthogonal sensing).

[Spectral Analysis of Dissolved Organic Matter of Tannery Wastewater in the Treatment Process].

PubMed

Fan, Chun-hui; Zhang, Ying-chao; Du, Bo; Song, Juan; Huai, Cui-qian; Wang, Jia-hong

2015-06-01

Tannery industry is one of the major traditional industries and important wastewater sources in China. The existing research mainly focus on the quality of inlet and outlet water, rather than the purification and transformation behavior of dissolved organic matter (DOM) in the treatment process of tannery wastewater. The UV spectra and fluorescence spectroscopy were used to detect the spectral characteristics of water samples in the treatment process, and it is analyzed that the formation process and the linear relationships between total fluorescence intensity and parameters. The results showed: the UV absorbance of DOM in wastewater increased firstly and then decreased with longer wavelength, and the wave peaks were found around the wavelength of 230 nr. The values of A253 /A203 and SUVA254 increased firstly and then decreased, indicating the complex reaction process related to free substituent and aromatic rings. The fluorescence peaks appeared at the regions of λ(ex/em) = 320-350/440- 460 and λ(ex/em) = 270-300/390-420, referred as visible humic-like and visible fulvic-like fluorescence, respectively. With the treatment process of tannery wastewater, the following fluorescence phenomenon were monitored, such as the blue-shift of humic-like fluorescence peak in the hydrolytic acidification tank, the appearance of tryptophan fluorescence peak in the second biochemical pond (λ(ex/em) = 290/340), the weak fluorescence peak in the fourth biochemical pond (λ(ex/em) = 350/520) and the stabilized fluorescence characteristics in the secondary sedimentation tank and water outlet. The achievements are helpful to investigate the degradation and formation behavior of water components, and significant for the fluorescence variation analysis in the treatment system. The removal rate of total fluorescence intensity of tannery wastewater fit better the removal rate of TOC with coefficient of r 0.835 5. The UV spectra and 3D-EEMs are effective to reveal the purification behavior and mechanism of tannery wastewater.

Light-Induced Fluorescence Modulation of Quantum Dot-Crystal Violet Conjugates: Stochastic Off-On-Off Cycles for Multicolor Patterning and Super-Resolution.

PubMed

Jung, Sungwook; Park, Joonhyuck; Bang, Jiwon; Kim, Jae-Yeol; Kim, Cheolhee; Jeon, Yongmoon; Lee, Seung Hwan; Jin, Ho; Choi, Sukyung; Kim, Bomi; Lee, Woo Jin; Pack, Chan-Gi; Lee, Jong-Bong; Lee, Nam Ki; Kim, Sungjee

2017-06-07

Photoswitching or modulation of quantum dots (QDs) can be promising for many fields that include display, memory, and super-resolution imaging. However, such modulations have mostly relied on photomodulations of conjugated molecules in QD vicinity, which typically require high power of high energy photons at UV. We report a visible light-induced facile modulation route for QD-dye conjugates. QD crystal violets conjugates (QD-CVs) were prepared and the crystal violet (CV) molecules on QD quenched the fluorescence efficiently. The fluorescence of QD-CVs showed a single cycle of emission burst as they go through three stages of (i) initially quenched "off" to (ii) photoactivated "on" as the result of chemical change of CVs induced by photoelectrons from QD and (iii) back to photodarkened "off" by radical-associated reactions. Multicolor on-demand photopatterning was demonstrated using QD-CV solid films. QD-CVs were introduced into cells, and excitation with visible light yielded photomodulation from "off" to "on" and "off" by nearly ten fold. Individual photoluminescence dynamics of QD-CVs was investigated using fluorescence correlation spectroscopy and single QD emission analysis, which revealed temporally stochastic photoactivations and photodarkenings. Exploiting the stochastic fluorescence burst of QD-CVs, simultaneous multicolor super-resolution localizations were demonstrated.

A Biocompatible and Biodegradable Protein Hydrogel with Green and Red Autofluorescence: Preparation, Characterization and In Vivo Biodegradation Tracking and Modeling

PubMed Central

Ma, Xiaoyu; Sun, Xiangcheng; Hargrove, Derek; Chen, Jun; Song, Donghui; Dong, Qiuchen; Lu, Xiuling; Fan, Tai-Hsi; Fu, Youjun; Lei, Yu

2016-01-01

Because of its good biocompatibility and biodegradability, albumins such as bovine serum albumin (BSA) and human serum albumin (HSA) have found a wide range of biomedical applications. Herein, we report that glutaraldehyde cross-linked BSA (or HSA) forms a novel fluorescent biological hydrogel, exhibiting new green and red autofluorescence in vitro and in vivo without the use of any additional fluorescent labels. UV-vis spectra studies, in conjunction with the fluorescence spectra studies including emission, excitation and synchronous scans, indicated that three classes of fluorescent compounds are presumably formed during the gelation process. SEM, FTIR and mechanical tests were further employed to investigate the morphology, the specific chemical structures and the mechanical strength of the as-prepared autofluorescent hydrogel, respectively. Its biocompatibility and biodegradability were also demonstrated through extensive in vitro and in vivo studies. More interestingly, the strong red autofluorescence of the as-prepared hydrogel allows for conveniently and non-invasively tracking and modeling its in vivo degradation based on the time-dependent fluorescent images of mice. A mathematical model was proposed and was in good agreement with the experimental results. The developed facile strategy to prepare novel biocompatible and biodegradable autofluorescent protein hydrogels could significantly expand the scope of protein hydrogels in biomedical applications. PMID:26813916

The EUSO program: Imaging of ultra-high energy cosmic rays by high-speed UV-video from space

NASA Astrophysics Data System (ADS)

Fuglesang, Christer; JEM-EUSO Collaboration

2017-11-01

The Extreme-Energy Cosmic Rays (EECR), with energy above 5•1019 eV, are very interesting objects to study that can provide new information about our universe. At the same time EECRs are exceptionally challenging to study because they are so rare. To obtain a reasonably large statistical sample, the JEM-EUSO collaboration aims to place a telescope into space. Various technologies are being developed and studied to achieve this goal. Several pathfinders are used for validation and testing. In particular, during 2017 a long-duration super-pressure balloon flight will observe the first high energy cosmic rays from above using the fluorescence technique, and a small test unit, Mini-EUSO, will be sent to ISS to measure the UV-background from Earth night side. In addition, these missions will provide various scientific results.

Optimization strategies for a fluorescent dye with bimodal excitation spectra: application to semiautomated proteomics

NASA Astrophysics Data System (ADS)

Patton, Wayne F.; Berggren, Kiera N.; Lopez, Mary F.

2001-04-01

Facilities engaged in proteome analysis differ significantly in the degree that they implement automated systems for high-throughput protein characterization. Though automated workstation environments are becoming more routine in the biotechnology and pharmaceutical sectors of industry, university-based laboratories often perform these tasks manually, submitting protein spots excised from polyacrylamide gels to institutional core facilities for identification. For broad compatibility with imaging platforms, an optimized fluorescent dye developed for proteomics applications should be designed taking into account that laser scanners use visible light excitation and that charge-coupled device camera systems and gas discharge transilluminators rely upon UV excitation. The luminescent ruthenium metal complex, SYPRO Ruby protein gel stain, is compatible with a variety of excitation sources since it displays intense UV (280 nm) and visible (470 nm) absorption maxima. Localization is achieved by noncovalent, electrostatic and hydrophobic binding of dye to proteins, with signal being detected at 610 nm. Since proteins are not covalently modified by the dye, compatibility with downstream microchemical characterization techniques such as matrix-assisted laser desorption/ionization-mass spectrometry is assured. Protocols have been devised for optimizing fluorophore intensity. SYPRO Ruby dye outperforms alternatives such as silver staining in terms of quantitative capabilities, compatibility with mass spectrometry and ease of integration into automated work environments.

Classroom Activity Connections: Lessons from Fluorescence

ERIC Educational Resources Information Center

MacCormac, Aoife; O'Brien, Emma; O'Kennedy, Richard

2010-01-01

This Classroom Activity Connections paper describes an extension to the "JCE" Classroom Activity #68 "Turning on the Light". A number of additional common items that display fluorescence under UV light are described, including fruits, vegetables, and seashells. Two classroom extensions on fluorescence are also described. From these activities,…

Factors that affect the EVA encapsulant discoloration rate upon accelerated exposure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pern, F.J.

1994-12-31

Several factors that may affect the net discoloration rate of the ethylene-vinyl acetate (EVA) copolymer encapsulants used in crystalline-Si photovoltaic (c-Si PV) modules upon accelerated exposure have been investigated by employing UV-visible spectrophotometry, spectrocolorimetry, and fluorescence analysis. A number of laminated films, including the two typical EVA formulations, A9918 and 15295, were studied. The results indicate that the rate of EVA discoloration is affected by the (1) curing agent and curing conditions; (2) presence and concentration of curing-generated, UV-excitable chromophores; (3) UV light intensity; (4) loss rate of the UV absorber, Cyasorb UV 531; (5) lamination; (6) film thickness; andmore » (7) photobleaching rate due to the diffusion of air into the laminated films. In general, the loss rate of the UV absorber and the rate of discoloration from light yellow to brown follow a sigmoidal pattern. A reasonable correlation for net changes in transmittance at 420 nm, yellowness index, and fluorescence peak area (or intensity) ratio is obtained as the extent of EVA discoloration progressed.« less

Computer-aided design of peptide near infrared fluorescent probe for tumor diagnosis

NASA Astrophysics Data System (ADS)

Zhang, Congying; Gu, Yueqing

2014-09-01

Integrin αvβ3 receptors are expressed on activated endothelial cells during neovascularization to maintain tumor growth, so they become hot research tagets in cancer diagnosis. Peptides possess several attractive features when compared to protein and small molecule, such as small size and high structural compatibility with target proteins. Efficient design of high-affinity peptide ligands to Integrin αvβ3 receptors has been an important problem. Designed peptides in silico provide a valuable and high-selectivity peptide, meanwhile decrease the time of drug screening. In this study, we design peptide which can bind with integrin αvβ3 via computer, and then synthesis near infrared fluorescent probe. The characterization of this near infrared fluorescent probe was detected by UV. To investigate the tumor cell targeting of this probe, it was labeled with visible fluorescent dye Rhodamine B (RhB) for microscopy. To evaluate the targeting capability of this near infrared fluorescent probe, mice bearing integrin αvβ3 positive tumor xenografts were used. In vitro cellular experiments indicated that this probe have a clear binding affinity to αvβ3-positive tumor cells. In vivo experiments confirmed the receptor binding specificity of this probe. The peptide of computational design can bind with integrin αvβ3. Combined peptide near-infrared fluorescent probe with imaging technology use for clinical and tumor diagnosis have a greater development in future.

[Two-photon up-conversion fluorescence of a neodymium organic framework Nd(BTC)].

PubMed

Xu, Hui; Jin, Run-zhi; Wu, Chun-yang; Yang, Yu; Qian, Guo-dong

2008-08-01

In the present work, a neodymium organic framework Nd(BTC) was synthesized by the solvothermal reaction of Nd(NO3)3 x 5H2O and H3BTC (BTC = 1,3,5-benzenetricarboxylate) in mixed solvents of DMF, ethanol and water, and was identified by elemental analysis. This MOF complex was characterized using X-ray diffraction (XRD), thermogravimetry-differential scanning calorimetry (TGA-DSC) analysis, UV-visible absorption spectra and fluorescence spectra. This polycrystalline powder lost DMF and H2O when heated to 120 degrees C, then remained stable till the temperature reached 500 degrees C. Near infrared fluorescence at 1064 nm due to the 4 F3/2--> 4 I11/2 transition of Nd3+ ions was exhibited when excited by 808 nm laser beam. It was also been found that up-conversion fluorescence of Nd(BTC) peaked at about 450 nm due to 2 D5/2 -->4 I11/2 transition of Nd3+ ions can be observed under the excitation of a 580 nm laser line. The mechanism of the up-conversion fluorescence of Nd(BTC) at around 450 nm under the excitation of 580 nm laser can be ascribed to both excited-state absorption and energy transfer up-conversion. This result indicated that such MOF can be employed as an up-conversion luminescence material in many potential application areas such as bio-labeling and fluorescence image.

Fluorescence Spectroscopic Properties of Normal and Abnormal Biomedical Materials

NASA Astrophysics Data System (ADS)

Pradhan, Asima

Steady state and time-resolved optical spectroscopy and native fluorescence is used to study the physical and optical properties occurring in diseased and non-diseased biological human tissue, in particular, cancer of the human breast, artery and the dynamics of a photosensitizer useful in photodynamic therapy. The main focus of the research is on the optical properties of cancer and atherosclerotic tissues as compared to their normal counterparts using the different luminescence based spectroscopic techniques such as steady state fluorescence, time-resolved fluorescence, excitation spectroscopy and phosphorescence. The excitation and steady-state spectroscopic fluorescence using visible excitation wavelength displays a difference between normal and malignant tissues. This difference is attributed to absorption of the emission by hemoglobin in normal tissues. This method using 488nm fails to distinguish neoplastic tissue such as benign tissues and tumors from malignant tumors. The time-resolved fluorescence at visible, near -uv and uv excitation wavelengths display non-exponential profiles which are significantly different for malignant tumors as compared to non-malignant tissues only with uv excitation. The differences observed with visible and near-uv excitation wavelengths are not as significant. The non-exponential profiles are interpreted as due to a combination of fluorophores along with the action of non-radiative processes. Low temperature luminescence studies confirm the occurrence of non-radiative decay processes while temporal studies of various relevant biomolecules indicate the probable fluorophores responsible for the observed signal in tissues. Phosphorescence from human tissues have been observed for the first time and lifetimes of a few hundred nanoseconds are measured for malignant and benign tissues. Time-resolved fluorescence studies of normal artery and atherosclerotic plaque have shown that a combination of two excitation wavelengths can distinguish fibrous and calcified atherosclerotic plaque from normal artery. A minor effort of the study involves the high intensity effects on the optical properties of the dye, doxycycline (a particular photosensitizer of the tetracycline group) occurring during relaxation when excited at different laser intensities. This study has been performed by observing the fluorescence lifetimes and quantum yields of DOTC at different excitation intensities. The results obtained support the sequential excited state absorption model.

Discrimination of corn from monocotyledonous weeds with ultraviolet (UV) induced fluorescence.

PubMed

Panneton, Bernard; Guillaume, Serge; Samson, Guy; Roger, Jean-Michel

2011-01-01

In production agriculture, savings in herbicides can be achieved if weeds can be discriminated from crop, allowing the targeting of weed control to weed-infested areas only. Previous studies demonstrated the potential of ultraviolet (UV) induced fluorescence to discriminate corn from weeds and recently, robust models have been obtained for the discrimination between monocots (including corn) and dicots. Here, we developed a new approach to achieve robust discrimination of monocot weeds from corn. To this end, four corn hybrids (Elite 60T05, Monsanto DKC 26-78, Pioneer 39Y85 (RR), and Syngenta N2555 (Bt, LL)) and four monocot weeds (Digitaria ischaemum (Schreb.) I, Echinochloa crus-galli (L.) Beauv., Panicum capillare (L.), and Setaria glauca (L.) Beauv.) were grown either in a greenhouse or in a growth cabinet and UV (327 nm) induced fluorescence spectra (400 to 755 nm) were measured under controlled or uncontrolled ambient light intensity and temperature. This resulted in three contrasting data sets suitable for testing the robustness of discrimination models. In the blue-green region (400 to 550 nm), the shape of the spectra did not contain any useful information for discrimination. Therefore, the integral of the blue-green region (415 to 455 nm) was used as a normalizing factor for the red fluorescence intensity (670 to 755 nm). The shape of the normalized red fluorescence spectra did not contribute to the discrimination and in the end, only the integral of the normalized red fluorescence intensity was left as a single discriminant variable. Applying a threshold on this variable minimizing the classification error resulted in calibration errors ranging from 14.2% to 15.8%, but this threshold varied largely between data sets. Therefore, to achieve robustness, a model calibration scheme was developed based on the collection of a calibration data set from 75 corn plants. From this set, a new threshold can be estimated as the 85% quantile on the cumulative frequency curve of the integral of the normalized red fluorescence. With this approach the classification error was nearly constant (16.0% to 18.5%), thereby indicating the potential of UV-induced fluorescence to reliably discriminate corn from monocot weeds.

Responses of photosynthetic properties and chloroplast ultrastructure of two moss crusts from a desert biological soil crust to supplementary UV-B radiation

NASA Astrophysics Data System (ADS)

Hui, Rong; Li, Xinrong; Zhao, Yang; Pan, Yanxia

2016-04-01

Our understanding of plant responses to supplementary ultraviolet-B (UV-B) radiation due to stratospheric ozone depletion has improved over recent decades. However, research on biological soil crusts (BSCs) is scarce and it remains controversial. Laboratory studies were conducted to investigate the influence of UV-B radiation on the Bryum argenteum and Didymodon vinealis isolated from BSCs, which are both dominant species in moss crusts found within patches of shrubs and herbs in the Tengger Desert of northern China. The aim of the current work was to evaluate whether supplementary UV-B radiation affected photosynthetic properties and chloroplast ultrastructure of two moss crusts and whether response differences were observed between the crusts. Four levels of UV-B radiation of 2.75 (control), 3.08, 3.25, and 3.41 W m-2 was achieved using fluorescence tube systems for 10 days, simulating 0, 6, 9, and 12% of stratospheric ozone at the latitude of Shapotou, respectively. We measured photosynthetic apparatus as assessed by chlorophyll a fluorescence parameters, photosynthetic pigment contents, and observations of chloroplast ultrastructure. Additionally, soluble proteins and UV-B absorbing compounds were simultaneously investigated. The results of this study showed that chlorophyll a fluorescence parameters (i.e., the maximal quantum yield of PSII photochemistry, the effective quantum yield of PSII photochemistry, and photochemical quenching coefficient), photosynthetic pigment contents, soluble protein contents, total flavonoid contents and the ultrastructure were negatively influenced by elevated UV-B radiation and the degree of detrimental effects significantly increased with the intensity of UV-B radiation. Moreover, results demonstrated that the negative effects on photosynthesis and chloroplast ultrastructure were more serious in B. argenteum than that in D. vinealis. These results may not only provide a potential mechanism for supplemental UV-B effects on photosynthesis of moss crusts, but also establish a theoretical basis for further studies of adaptation and response mechanisms of desert ecosystems under further ozone depletion.

Radiative lifetimes in B I using ultraviolet and vacuum-ultraviolet laser-induced fluorescence

NASA Technical Reports Server (NTRS)

O'Brian, T. R.; Lawler, J. E.

1992-01-01

Radiative lifetimes of the eight lowest even parity levels in the doublet system of B I are measured using time-resolved laser-induced fluorescence in the UV and VUV on an atomic beam of boron. The accurate lifetimes provide a base for improved determination of absolute transition probabilities in B I. The techniques described are broadly applicable to measurement of lifetimes of levels with transitions in the visible, UV, and VUV in almost any element.

Ultraviolet fluorescence to identify navel oranges with poor peel quality and decay

USDA-ARS?s Scientific Manuscript database

Navel oranges were sorted into four groups under ultraviolet (UV) illumination in commercial packinghouse black light rooms based upon the amount of fluorescence visible on each fruit to determine if fluorescence was predictive of peel quality. The groups corresponded to fruit with: 1) no fluorescen...

Contribution of glue layer into epidermis sample fluorescence dynamics

NASA Astrophysics Data System (ADS)

Salomatina, Elena V.; Chernova, Svetlana P.; Pravdin, Alexander B.

2000-04-01

In this work, the temporal behavior of autofluorescence of epidermis samples under UV-irradiation has ben studied. The samples were prepared using surface epidermis stripping technique. Fluorescence spectra and kinetic curves of fluorescence intensity have been obtained. It has been concluded that the glue composition used allows the measurement of epidermis fluorescence dynamics with the first 60 min of experiment.

Fluorescence and phosphorescence of photomultiplier window materials under electron irradiation

NASA Technical Reports Server (NTRS)

Viehmann, W.; Eubanks, A. G.; Bredekamp, J. H.

1974-01-01

The fluorescence and phosphorescence of photomultiplier window materials under electron irradiation were investigated using a Sr-90/Y-90 beta emitter as the electron source. Spectral emission curves of UV grade, optical grade, and electron-irradiated samples of MGF2 and LiF, CaF2, BaF2, sapphire, fused silica, and UV transmitting glasses were obtained over the spectral range of 200 nm to 650 nm. Fluorescence yields, expressed as the number of counts in a solid angle of 2 pi steradian per 1MeV of incident electron energy deposited, were determined on these materials utilizing photomultiplier tubes with cesium telluride, bialkali, and trialkali (S-20) photocathodes, respectively.

Fluorimetric study on the interaction between Norfloxacin and Proflavine hemisulphate.

PubMed

More, Vishalkumar R; Anbhule, Prashant V; Lee, Sang H; Patil, Shivajirao R; Kolekar, Govind B

2011-07-01

The interaction between Norfloxacin (NF) and Proflavine hemisulphate (PF) was investigated by spectroscopic tools like UV-VIS absorption and Fluorescence spectroscopy. It was proved that fluorescence quenching of NF by PF is due to the formation of NF-PF complex which was supported by UV-VIS absorption study. The study of thermodynamic parameters suggested that the key interacting forces are hydrogen bond and van der Waal's interactions and the binding interaction was spontaneous. The distance r between NF and PF was obtained according to the Förster's theory of non-radiative energy transfer. The fluorescence quenching mechanism was applied to estimate PF directly from pharmaceutical samples. © Springer Science+Business Media, LLC 2011

Skin tumor development after UV irradiation and photodynamic therapy is unaffected by short-term pretreatment with 5-fluorouracil, imiquimod and calcipotriol. An experimental hairless mouse study.

PubMed

Bay, Christiane; Togsverd-Bo, Katrine; Lerche, Catharina M; Haedersdal, Merete

2016-01-01

Photodynamic therapy (PDT) delays ultraviolet (UV) radiation-induced squamous cell carcinomas (SCCs) in hairless mice. Efficacy may be enhanced by combining PDT with antineoplastic or pro-differentiating agents. We investigated if pretreatment with 5-fluorouracil (5FU), imiquimod (IMIQ) or calcipotriol (CAL) before PDT further delays tumor onset. Hairless mice (n=224) were exposed 3 times weekly to 3 standard erythema doses (SED) of UV radiation. Methyl-aminolevulinate (MAL)-PDT sessions were given on days 45 and 90 before SCC development. Three applications of topical 5FU, IMIQ or CAL were given before each PDT session. Fluorescence photography quantified protoporphyrin IX (PpIX) formation. PDT delayed UV-induced SCC development by 59 days (212 days UV-MAL-PDT vs. 153 days UV-control, P<0.001). Pretreatment with 5FU, IMIQ or CAL before PDT did not further delay SCC onset compared to PDT alone (207 days UV-5FU-MAL-PDT, 215 days UV-IMIQ-MAL-PDT, 206 days UV-CAL-MAL-PDT vs. 212 days UV-MAL-PDT, P=ns). PpIX fluorescence intensified by 5FU-pretreatment (median 21,392 au UV-5FU-MAL-PDT, P=0.011), decreased after IMIQ-pretreatment (12,452 au UV-IMIQ-MAL-PDT, P<0.001), and was unaffected by CAL-pretreatment (19,567 au UV-CAL-MAL-PDT, P=ns) compared to MAL alone (18,083 au UV-MAL-PDT). Short-term three-day pretreatment with 5FU, IMIQ and CAL before PDT does not further delay tumor onset in UV-exposed hairless mice. Copyright © 2015 Elsevier B.V. All rights reserved.

Coordinated in Situ Analyses of Organic Nanoglobules in the Sutter's Mill Meteorite

NASA Technical Reports Server (NTRS)

Nakamura-Messenger, K.; Messenger, S.; Keller, L. P.; Clemett, S. J.; Nguyen, A. N.; Gibson, E. K.

2013-01-01

The Sutter's Mill meteorite is a newly fallen carbonaceous chondrite that was collected and curated quickly after its fall. Preliminary petrographic and isotopic investigations suggest affinities to the CM2 carbonaceous chondrites. The primitive nature of this meteorite and its rapid recovery provide an opportunity to investigate primordial solar system organic matter in a unique new sample. Here we report in-situ analyses of organic nanoglobules in the Sutter's Mill meteorite using UV fluorescence imaging, Fourier-transform infrared spectroscopy (FTIR), scanning transmission electron microscopy (STEM), NanoSIMS, and ultrafast two-step laser mass spectrometry (ultra-L2MS).

The damaging effect of UV-C irradiation on lens alpha-crystallin.

PubMed

Fujii, Noriko; Uchida, Hiroki; Saito, Takeshi

2004-11-02

To evaluate the effect of UV-C irradiation on the structural properties of alpha-crystallin and its chaperone activity. alpha- and betaL-crystallins were isolated from bovine lenses using gel chromatography. The purified alpha-crystallin was subjected to UV-C irradiation (254 nm; 1, 2, 5, 10, 20, 50 J/cm2). We measured the tryptophan fluorescence, circular dichroism (CD) spectroscopy in the far UV, and the chaperone activity of both irradiated and non-irradiated alpha-crystallin. The tryptophan fluorescence of alpha-crystallin decreased, whereas the N-formylkynurenine fluorescence increased markedly with increasing doses of UV-C irradiation. Both the oxidation of Met1 and the racemization of Asp151 of alphaA-crystallin increased at a dose of 1-2 J/cm2 and then gradually decreased. The CD spectrum showed that the secondary structure of alpha-crystallin altered with increasing radiation dose, and almost all of the beta-sheet structure was lost at doses above 50 J/cm2. The chaperone activity of alpha-crystallin irradiated with doses under 5 J/cm2 remained intact. However, it was reduced to only 40% after irradiation at 10 J/cm2. Our study suggests that photo-oxidation of tryptophan residues in alpha-crystallin may be one of the events that affects the three-dimensional packing array and chaperone activity of this lens protein.

Characterizing the Interaction between tartrazine and two serum albumins by a hybrid spectroscopic approach.

PubMed

Pan, Xingren; Qin, Pengfei; Liu, Rutao; Wang, Jing

2011-06-22

Tartrazine is an artificial azo dye commonly used in food products. The present study evaluated the interaction of tartrazine with two serum albumins (SAs), human serum albumin (HSA) and bovine serum albumin (BSA), under physiological conditions by means of fluorescence, three-dimensional fluorescence, UV-vis absorption, and circular dichroism (CD) techniques. The fluorescence data showed that tartrazine could bind to the two SAs to form a complex. The binding process was a spontaneous molecular interaction procedure, in which van der Waals and hydrogen bond interactions played a major role. Additionally, as shown by the UV-vis absorption, three-dimensional fluorescence, and CD results, tartrazine could lead to conformational and some microenvironmental changes of both SAs, which may affect the physiological functions of SAs. The work provides important insight into the mechanism of toxicity of tartrazine in vivo.

Ascorbic acid as a free radical scavenger in porcine and bovine aqueous humour.

PubMed

Erb, Carl; Nau-Staudt, Kerstin; Flammer, Josef; Nau, Werner

2004-01-01

To study the antioxidant activity, UV absorption, concentration and stability of ascorbic acid (AA) in porcine and bovine aqueous humour (AH). Porcine and bovine AH was taken within 5 min after death and frozen at -70 degrees C. The characteristic UV absorption band of AA and the concentration of AA in AH was determined by UV spectrophotometry. The antioxidant activity of AA to serve as a free radical scavenger in AH has been determined by using a novel fluorescent probe for antioxidants, the azoalkane 2,3-diazabicyclo[2.2.2]oct-2-ene (DBO). The fluorescence lifetime and intensity of this probe reflect the concentration of dissolved antioxidants. The time-resolved fluorescence of DBO (laser excitation at 351 nm) in AH and in a neutral phosphate-buffered saline (PBS) solution containing only the natural amount of AA as an additive were measured. The characteristic UV absorption band of AA has its maximum at 266 nm in AH. The concentration of AA in porcine and bovine AH was found to be 0.547 +/- 0.044 and 1.09 +/- 0.16 mM, respectively, by spectrophotometry. The fluorescence lifetime of the probe DBO was reduced from 320 +/- 5 ns in pure aerated PBS to 205 +/- 5 ns in porcine AH and 165 +/- 3 ns in bovine AH. A detailed kinetic analysis of the lifetime shortening suggests that AA contributes approximately 75 and 85% to the antioxidant activity of porcine and bovine AH, respectively. Our experiments suggest that AA is the major contributor to the antioxidant activity of porcine and bovine AH. The role of AA to serve as an antioxidant in AH is discussed. In addition, UV spectrophotometry is established as an alternative method to determine the concentration of AA in AH. Copyright 2004 S. Karger AG, Basel

PARAFAC Modeling of Irradiation- and Oxidation-Induced Changes in Fluorescent Dissolved Organic Matter Extracted from Poultry Litter.

PubMed

Mangalgiri, Kiranmayi P; Timko, Stephen A; Gonsior, Michael; Blaney, Lee

2017-07-18

Parallel factor analysis (PARAFAC) applied to fluorescence excitation emission matrices (EEMs) allows quantitative assessment of the composition of fluorescent dissolved organic matter (DOM). In this study, we fit a four-component EEM-PARAFAC model to characterize DOM extracted from poultry litter. The data set included fluorescence EEMs from 291 untreated, irradiated (253.7 nm, 310-410 nm), and oxidized (UV-H 2 O 2 , ozone) poultry litter extracts. The four components were identified as microbial humic-, terrestrial humic-, tyrosine-, and tryptophan-like fluorescent signatures. The Tucker's congruence coefficients for components from the global (i.e., aggregated sample set) model and local (i.e., single poultry litter source) models were greater than 0.99, suggesting that the global EEM-PARAFAC model may be suitable to study poultry litter DOM from individual sources. In general, the transformation trends of the four fluorescence components were comparable for all poultry litter sources tested. For irradiation at 253.7 nm, ozonation, and UV-H 2 O 2 advanced oxidation, transformation of the humic-like components was slower than that of the tryptophan-like component. The opposite trend was observed for irradiation at 310-410 nm, due to differences in UV absorbance properties of components. Compared to the other EEM-PARAFAC components, the tyrosine-like component was fairly recalcitrant in irradiation and oxidation processes. This novel application of EEM-PARAFAC modeling provides insight into the composition and fate of agricultural DOM in natural and engineered systems.

Assessment of organic pollution of an industrial river by synchronous fluorescence and UV-vis spectroscopy: the Fensch River (NE France).

PubMed

Assaad, Aziz; Pontvianne, Steve; Pons, Marie-Noëlle

2017-05-01

To rapidly monitor the surface water quality in terms of organic pollution of an industrial river undergoing restoration, optical methods (UV-visible spectrometry and fluorescence) were applied in parallel to classical physical-chemical analyses. UV-visible spectra were analyzed using the maximum of the second derivative at 225 nm (related to nitrates), specific absorbance at 254 nm (SUVA 254 ), and the spectral slope between 275 and 295 nm (S 275-295 ) (related to the aromaticity and molecular weight of dissolved organic carbon). The synchronous fluorescence spectra (wavelength difference = 50 nm) exhibited a high variability in the composition of dissolved organic material between the upstream and downstream sections and also versus time. The principal components analysis of the entire set of synchronous fluorescence spectra helped to define three river sections with different pollution characteristics. Spectral decomposition was applied to the two most upstream sections: five fluorophores, classical in rivers impacted by domestic sewage and related to protein-like (λ ex  = 280 nm) and humic-like fluorescence (M-type with λ ex  ≈ 305-310 nm and C-type with λ ex  ≥ 335 nm), were identified. The irregular shape of the synchronous fluorescence spectra in the most downstream section is likely due to organic pollutants of industrial origin; however, their variability and the complexity of the spectra did not allow the further elucidation of their nature.

Hydrophilic interaction liquid chromatography of anthranilic acid-labelled oligosaccharides with a 4-aminobenzoic acid ethyl ester-labelled dextran hydrolysate internal standard.

PubMed

Neville, David C A; Alonzi, Dominic S; Butters, Terry D

2012-04-13

Hydrophilic interaction liquid chromatography (HILIC) of fluorescently labelled oligosaccharides is used in many laboratories to analyse complex oligosaccharide mixtures. Separations are routinely performed using a TSK gel-Amide 80 HPLC column, and retention times of different oligosaccharide species are converted to glucose unit (GU) values that are determined with reference to an external standard. However, if retention times were to be compared with an internal standard, consistent and more accurate GU values would be obtained. We present a method to perform internal standard-calibrated HILIC of fluorescently labelled oligosaccharides. The method relies on co-injection of 4-aminobenzoic acid ethyl ester (4-ABEE)-labelled internal standard and detection by UV absorption, with 2-AA (2-aminobenzoic acid)-labelled oligosaccharides. 4-ABEE is a UV chromophore and a fluorophore, but there is no overlap of the fluorescent spectrum of 4-ABEE with the commonly used fluorescent reagents. The dual nature of 4-ABEE allows for accurate calculation of the delay between UV and fluorescent signals when determining the GU values of individual oligosaccharides. The GU values obtained are inherently more accurate as slight differences in gradients that can influence retention are negated by use of an internal standard. Therefore, this paper provides the first method for determination of HPLC-derived GU values of fluorescently labelled oligosaccharides using an internal calibrant. Copyright © 2012 Elsevier B.V. All rights reserved.

Quantitation of polymethoxylated flavones in orange juice by high-performance liquid chromatography.

PubMed

Rouseff, R L; Ting, S V

1979-08-01

A quantitative high-performance liquid chromatographic (HPLC) procedure for the determination of the five major polymethoxylated flavones (PMFs) in orange juice has been developed. It employs a unique ternary solvent system with coupled UV-fluorescence detection. The dual detectors were employed to determine the presence of interfering substances and served as a cross check on quantitation. Stop flow UV and fluorescence scanning was used to identify peaks and determine the presence of impurities. Although all five citrus PMFs fluoresce, some HPLC fluorescence peaks were too small to be of much practical use. All five citrus PMFs could be quantitated satisfactorily with the fixed wavelength UV (313 nm) detector. The HPLC procedure has been used to evaluate each step in the preparation. The optimum extracting solvent was selected and one time consuming step was eliminated, as it was found to be unnecessary. HPLC values for nobiletin and sinensetin are in good agreement with the thin-layer chromatographic (TLC) values in the literature. HPLC values for the other three flavones were considerably lower than those reported in the literature. The HPLC procedure is considerably faster than the TLC procedure with equal or superior precision and accuracy.

Effect of metal ions on autofluorescence of the dry rot fungus Serpula lacrymans grown on spruce wood.

PubMed

Gabriel, Jiří; Žižka, Zdeněk; Švec, Karel; Nasswettrová, Andrea; Šmíra, Pavel; Kofroňová, Olga; Benada, Oldřich

2016-03-01

This work describes autofluorescence of the mycelium of the dry rot fungus Serpula lacrymans grown on spruce wood blocks impregnated with various metals. Live mycelium, as opposed to dead mycelium, exhibited yellow autofluorescence upon blue excitation, blue fluorescence with ultraviolet (UV) excitation, orange-red and light-blue fluorescence with violet excitation, and red fluorescence with green excitation. Distinctive autofluorescence was observed in the fungal cell wall and in granula localized in the cytoplasm. In dead mycelium, the intensity of autofluorescence decreased and the signal was diffused throughout the cytoplasm. Metal treatment affected both the color and intensity of autofluorescence and also the morphology of the mycelium. The strongest yellow signal was observed with blue excitation in Cd-treated samples, in conjunction with increased branching and the formation of mycelial loops and protrusions. For the first time, we describe pink autofluorescence that was observed in Mn-, Zn-, and Cu-treated samples with UV, violet or. blue excitation. The lowest signals were obtained in Cu- and Fe-treated samples. Chitin, an important part of the fungal cell wall exhibited intensive primary fluorescence with UV, violet, blue, and green excitation.

Electrochemical vapor generation of selenium species after online photolysis and reduction by UV-irradiation under nano TiO2 photocatalysis and its application to selenium speciation by HPLC coupled with atomic fluorescence spectrometry.

PubMed

Liang, Jing; Wang, Qiuquan; Huang, Benli

2005-01-01

An online UV photolysis and UV/TiO2 photocatalysis reduction device (UV-UV/TiO2 PCRD) and an electrochemical vapor generation (ECVG) cell have been used for the first time as an interface between high-performance liquid chromatography (HPLC) and atomic fluorescence spectrometry (AFS) for selenium speciation. The newly designed ECVG cell of approximately 115 microL dead volume consists of a carbon fiber cathode and a platinum loop anode; the atomic hydrogen generated on the cathode was used to reduce selenium to vapor species for AFS determination. The noise was greatly reduced compared with that obtained by use of the UV-UV/TiO2 PCRD-KBH4-acid interface. The detection limits obtained for seleno-DL: -cystine (SeCys), selenite (Se(IV)), seleno-DL: -methionine (SeMet), and selenate (Se(VI)) were 2.1, 2.9, 4.3, and 3.5 ng mL(-1), respectively. The proposed method was successfully applied to the speciation of selenium in water-soluble extracts of garlic shoots cultured with different selenium species. The results obtained suggested that UV-UV/TiO2 PCRD-ECVG should be an effective interface between HPLC and AFS for the speciation of elements amenable to vapor generation, and is superior to methods involving KBH4.

Measurement of pyrimidine (6-4) photoproducts in DNA by a mild acidic hydrolysis-HPLC fluorescence detection assay.

PubMed

Douki, T; Voituriez, L; Cadet, J

1995-03-01

Pyrimidine (6-4) pyrimidone photoproducts constitute one of the major classes of DNA lesions induced by far-UV irradiation. However, their biological role remains difficult to assess partly because of the lack of a specific and sensitive assay for monitoring their formation in DNA. Here is presented a measurement method based on the release of the (6-4) base adducts from DNA followed by an HPLC separation associated with a sensitive and specific fluorescence detection. The quantitative and mechanistic aspects of the chemical hydrolysis, based on the use of hydrogen fluoride stabilized in pyridine, were investigated, using dinucleoside monophosphate (6-4) photoproducts as model compounds. The final hydrolysis products were isolated and characterized by UV, fluorescence, mass, and 1H NMR spectroscopies. Application of the assay to far-UV irradiated calf thymus DNA provided information on the sequence effect on the rate of formation of three of the four possible bipyrimidine (6-4) photoproducts.

A new on-axis micro-spectrophotometer for combining Raman, fluorescence and UV/Vis absorption spectroscopy with macromolecular crystallography at the Swiss Light Source

PubMed Central

Pompidor, Guillaume; Dworkowski, Florian S. N.; Thominet, Vincent; Schulze-Briese, Clemens; Fuchs, Martin R.

2013-01-01

The combination of X-ray diffraction experiments with optical methods such as Raman, UV/Vis absorption and fluorescence spectroscopy greatly enhances and complements the specificity of the obtained information. The upgraded version of the in situ on-axis micro-spectrophotometer, MS2, at the macromolecular crystallography beamline X10SA of the Swiss Light Source is presented. The instrument newly supports Raman and resonance Raman spectroscopy, in addition to the previously available UV/Vis absorption and fluorescence modes. With the recent upgrades of the spectral bandwidth, instrument stability, detection efficiency and control software, the application range of the instrument and its ease of operation were greatly improved. Its on-axis geometry with collinear X-ray and optical axes to ensure optimal control of the overlap of sample volumes probed by each technique is still unique amongst comparable facilities worldwide and the instrument has now been in general user operation for over two years. PMID:23955041

A new on-axis micro-spectrophotometer for combining Raman, fluorescence and UV/Vis absorption spectroscopy with macromolecular crystallography at the Swiss Light Source.

PubMed

Pompidor, Guillaume; Dworkowski, Florian S N; Thominet, Vincent; Schulze-Briese, Clemens; Fuchs, Martin R

2013-09-01

The combination of X-ray diffraction experiments with optical methods such as Raman, UV/Vis absorption and fluorescence spectroscopy greatly enhances and complements the specificity of the obtained information. The upgraded version of the in situ on-axis micro-spectrophotometer, MS2, at the macromolecular crystallography beamline X10SA of the Swiss Light Source is presented. The instrument newly supports Raman and resonance Raman spectroscopy, in addition to the previously available UV/Vis absorption and fluorescence modes. With the recent upgrades of the spectral bandwidth, instrument stability, detection efficiency and control software, the application range of the instrument and its ease of operation were greatly improved. Its on-axis geometry with collinear X-ray and optical axes to ensure optimal control of the overlap of sample volumes probed by each technique is still unique amongst comparable facilities worldwide and the instrument has now been in general user operation for over two years.

Giardia duodenalis: Number and Fluorescence Reduction Caused by the Advanced Oxidation Process (H2O2/UV)

PubMed Central

Guimarães, José Roberto; Franco, Regina Maura Bueno; Guadagnini, Regiane Aparecida; dos Santos, Luciana Urbano

2014-01-01

This study evaluated the effect of peroxidation assisted by ultraviolet radiation (H2O2/UV), which is an advanced oxidation process (AOP), on Giardia duodenalis cysts. The cysts were inoculated in synthetic and surface water using a concentration of 12 g H2O2 L−1 and a UV dose (λ = 254 nm) of 5,480 mJcm−2. The aqueous solutions were concentrated using membrane filtration, and the organisms were observed using a direct immunofluorescence assay (IFA). The AOP was effective in reducing the number of G. duodenalis cysts in synthetic and surface water and was most effective in reducing the fluorescence of the cyst walls that were present in the surface water. The AOP showed a higher deleterious potential for G. duodenalis cysts than either peroxidation (H2O2) or photolysis (UV) processes alone. PMID:27379301

Novel selective and sensitive optical chemosensor based on phenylfluorone derivative for detection of Ge(IV) ion in aqueous solution.

PubMed

Keawwangchai, Somchai; Morakot, Nongnit; Keawwangchai, Tasawan

2018-09-05

A water soluble chemosensor for Ge 4+ ion based on fluorone derivative containing 3,4-bis(2-(diethylamino)-2-oxoethoxy)phenyl (R8) has been synthesized. The binding abilities between R8 and 10 equiv. of Na + , K + , Ca 2+ , Fe 2+ , Cu 2+ , Cd 2+ , Hg 2+ , Pb 2+ , Al 3+ , Cr 3+ , Fe 3+ and Ge 4+ ions in 1% v/v EtOH-water (tris-buffer pH 7.0) were studied using UV-vis and fluorescence spectrophotometry. When observed by naked-eyes, the color of R8 changed from yellow-orange to pink and the fluorescent color changed from green to non-fluorescence when complexed with Ge 4+ ion. The spectral analysis showed that UV-vis absorption and fluorescence emission intensity of R8 decreased dramatically when Ge 4+ ion was added comparing with other ions. To explain this behavior, the quantum calculation was performed using the hybrid density functional at B3LYP /LanL2DZ level of theory. The calculated orbital energies indicated that the decreasing of UV-vis absorption and the quenching of fluorescence were due to the complexation induced metal to ligand charge transfer. The association constants (K a ) of R8-Ge 4+ complexes calculated from Benesi-Hildebrand equation was 6.21 × 10 5  M -1 . The UV-vis detection limit for Ge 4+ was 4.40 × 10 -7  M which was three orders of magnitude lower than those of Al 3+ , Cd 2+ , Cu 2+ and Na + ion. Copyright © 2018 Elsevier B.V. All rights reserved.

Attaching quantum dots to HER2 specific phage antibodies

NASA Astrophysics Data System (ADS)

Chu, Viet Ha; Nghiem, Thi Ha Lien; Huyen La, Thi; Dieu Thuy Ung, Thi; Huan Le, Quang; Thuan Tong, Kim; Liem Nguyen, Quang; Nhung Tran, Hong

2010-06-01

This work presents the results of the attachment of Qdot 655 ITKTM amino (PEG) quantum dots (QDs) (Invitrogen) and CdTe QDs (provided by Institute of Materials Science, VAST) to HER2 (Human Epidermal growth factor Receptor 2) specific phage antibodies (Abs) (provided by Institute of Biotechnology, VAST) in solution. The QDs were attached to the phage display specific HER2 Abs to form a complex QD-Ab. The QDs and complex QD-Ab were characterized by UV-VIS spectroscopy, transmission electron microscopy (TEM) and fluorescence microscopy. The fluorescence images show the QDs conjugated to the phage. Due to the QDs attaching to the surface, the phage dimensions were amplified, so its shape could be observed by optical microscopy. The complex QD-Ab was stable and lasted for a month. The results illustrate the value of the HER2 phage-QD complex as a cancer detection platform.

Influence of reaction time and synthesis temperature on the physical properties of ZnO nanoparticles synthesized by the hydrothermal method

NASA Astrophysics Data System (ADS)

Wasly, H. S.; El-Sadek, M. S. Abd; Henini, Mohamed

2018-01-01

Influence of synthesis temperature and reaction time on the structural and optical properties of ZnO nanoparticles synthesized by the hydrothermal method was investigated using X-ray diffraction (XRD), high resolution transmission electron microscopy (HR-TEM), energy-dispersive X-ray, Fourier transform infra-red spectroscopy, and UV-visible and fluorescence spectroscopy. The XRD pattern and HR-TEM images confirmed the presence of crystalline hexagonal wurtzite ZnO nanoparticles with average crystallite size in the range 30-40 nm. Their energy gap determined by fluorescence was found to depend on the synthesis temperature and reaction time with values in the range 2.90-3.78 eV. Thermal analysis, thermogravimetric and the differential scanning calorimetry were used to study the thermal reactions and weight loss with heat of the prepared ZnO nanoparticles.

Isolation and Characterization of Precise Dye/Dendrimer Ratios

PubMed Central

Dougherty, Casey A.; Furgal, Joseph C.; van Dongen, Mallory A.; Goodson, Theodore; Banaszak Holl, Mark M.; Manono, Janet; DiMaggio, Stassi

2014-01-01

Fluorescent dyes are commonly conjugated to nanomaterials for imaging applications using stochastic synthesis conditions that result in a Poisson distribution of dye/particle ratios and therefore a broad range of photophysical and biodistribution properties. We report the isolation and characterization of generation 5 poly(amidoamine) (G5 PAMAM) dendrimer samples containing 1, 2, 3, and 4 fluorescein (FC) or 6-carboxytetramethylrhodamine succinimidyl ester (TAMRA) dyes per polymer particle. For the fluorescein case, this was achieved by stochastically functionalizing dendrimer with a cyclooctyne `click' ligand, separation into sample containing precisely defined `click' ligand/particle ratios using reverse-phase high performance liquid chromatography (rp-HPLC), followed by reaction with excess azide-functionalized fluorescein dye. For the TAMRA samples, stochastically functionalized dendrimer was directly separated into precise dye/particle ratios using rp-HPLC. These materials were characterized using 1H and 19F NMR, rp-HPLC, UV-Vis and fluorescence spectroscopy, lifetime measurements, and MALDI. PMID:24604830

Interactions of benzotriazole UV stabilizers with human serum albumin: Atomic insights revealed by biosensors, spectroscopies and molecular dynamics simulations.

PubMed

Zhuang, Shulin; Wang, Haifei; Ding, Keke; Wang, Jiaying; Pan, Liumeng; Lu, Yanli; Liu, Qingjun; Zhang, Chunlong

2016-02-01

Benzotriazole UV stabilizers (BZTs) belong to one prominent group of ultraviolet (UV) stabilizers and are widely used in various plastics materials. Their large production volumes, frequent detections in the environment and potential toxicities have raised increasing public concern. BZTs can be transported in vivo by transport proteins in plasma and the binding association to transport proteins may serve as a significant parameter to evaluate the bioaccumulative potential. We utilized a novel HSA biosensor, circular dichroism spectroscopy, fluorescence spectroscopy to detect the dynamic interactions of six BZTs (UV-326, UV-327, UV-328, UV-329, UV-P, and BZT) with human serum albumin (HSA), and characterized the corresponding structure-activity relationships (SAR) by molecular dynamics simulations. All test BZTs potently bind at Sudlow site I of HSA with a binding constant of 10(4) L/mol at 298 K. Minor changes in the moieties of BZTs affect their interactions with HSA and differently induce conformations of HSA. Their binding reduced electrochemical impedance spectra and α-helix content of HSA, caused slight red-shifted emission, and changed fluorescence lifetime components of HSA in a concentration-dependent mode. UV-327 and UV-329 form hydrogen bonds with HSA, while UV-329, UV-P and BZT bind HSA with more favorable electrostatic interactions. Our in vitro and in silico study offered a significant framework toward the understanding of risk assessment of BZTs and provides guide for future design of environmental benign BZTs-related materials. Copyright © 2015 Elsevier Ltd. All rights reserved.

Alcohol-induced versus anion-induced states of alpha-chymotrypsinogen A at low pH.

PubMed

Khan, F; Khan, R H; Muzammil, S

2000-09-29

Characterization of conformational transition and folding intermediates is central to the study of protein folding. We studied the effect of various alcohols (trifluoroethanol (TFE), butanol, propanol, ethanol and methanol) and salts (K(3)FeCN(6), Na(2)SO(4), KClO(4) and KCl) on the acid-induced state of alpha-chymotrypsinogen A, a predominantly beta-sheet protein, at pH 2.0 by near-UV circular dichroism (CD), far-UV CD and 1-anilinonaphthalene-8-sulfonic acid (ANS) fluorescence measurements. Addition of alcohols led to an increase in ellipticity value at 222 nm indicating the formation of alpha-helical structure. The order of effectiveness of alcohols was shown to be TFE>butanol>propanol>ethanol>methanol. ANS fluorescence data showed a decrease in fluorescence intensity on alcohol addition, suggesting burial of hydrophobic patches. The near-UV CD spectra showed disruption of tertiary structure on alcohol addition. No change in ellipticity was observed on addition of salts at pH 2.0, whereas in the presence of 2 M urea, salts were found to induce a molten globule-like state as evident from the increases in ellipticity at 222 nm and ANS fluorescence indicating exposure of hydrophobic regions of the protein. The effectiveness in inducing the molten globule-like state, i.e. both increase in ellipticity at 222 nm and increase in ANS fluorescence, followed the order K(3)FeCN(6)>Na(2)SO(4)>KClO(4)>KCl. The loss of signal in the near-UV CD spectrum on addition of alcohols indicating disordering of tertiary structure results suggested that the decrease in ANS fluorescence intensity may be attributed to the unfolding of the ANS binding sites. The results imply that the alcohol-induced state had characteristics of an unfolded structure and lies between the molten globule and the unfolded state. Characterization of such partially folded states has important implications for protein folding.

Visualizing and quantifying dose distribution in a UV reactor using three-dimensional laser-induced fluorescence.

PubMed

Gandhi, Varun N; Roberts, Philip J W; Kim, Jae-Hong

2012-12-18

Evaluating the performance of typical water treatment UV reactors is challenging due to the complexity in assessing spatial and temporal variation of UV fluence, resulting from highly unsteady, turbulent nature of flow and variation in UV intensity. In this study, three-dimensional laser-induced fluorescence (3DLIF) was applied to visualize and quantitatively analyze a lab-scale UV reactor consisting of one lamp sleeve placed perpendicular to flow. Mapping the spatial and temporal fluence delivery and MS2 inactivation revealed the highest local fluence in the wake zone due to longer residence time and higher UV exposure, while the lowest local fluence occurred in a region near the walls due to short-circuiting flow and lower UV fluence rate. Comparing the tracer based decomposition between hydrodynamics and IT revealed similar coherent structures showing the dependency of fluence delivery on the reactor flow. The location of tracer injection, varying the height and upstream distance from the lamp center, was found to significantly affect the UV fluence received by the tracer. A Lagrangian-based analysis was also employed to predict the fluence along specific paths of travel, which agreed with the experiments. The 3DLIF technique developed in this study provides new insight on dose delivery that fluctuates both spatially and temporally and is expected to aid design and optimization of UV reactors as well as validate computational fluid dynamics models that are widely used to simulate UV reactor performances.

UV plasmonic enhancement through three dimensional nano-cavity antenna array in aluminum

NASA Astrophysics Data System (ADS)

Mao, Jieying; Stevenson, Peter; Montanaric, Danielle; Wang, Yunshan; Shumaker-Parry, Jennifer S.; Harris, Joel M.; Blair, Steve

2017-08-01

Metallic nanostructure can enhance fluorescence through excited surface plasmons which increase the local field as well as improve its quantum efficiency. When coupling to cavity resonance with proper gap dimension, gap hot spots can be generated to interact with fluorescence at their excitation/emission region in UV. A 3D nano-cavity antenna array in Aluminum has been conducted to generate local hot spot resonant at fluorescence emission resonance. Giant field enhancement has been achieved through coupling fundamental resonance modes of nanocavity into surface plasmons polaritons (SPPs). In this work, two distinct plasmonic structure of 3D resonant cavity nanoantenna has been studied and its plasmonic response has been scaled down to the UV regime through finite-difference-time-domain (FDTD) method. Two different strategies for antenna fabrication will be conducted to obtain D-coupled Dots-on-Pillar Antenna array (D2PA) through Focus Ion Beam (FIB) and Cap- Hole Pair Antenna array (CHPA) through nanosphere template lithography (NTL). With proper optimization of the structures, D2PA and CHPA square array with 280nm pitch have achieved distinct enhancement at fluorophore emission wavelength 350nm and excitation wavelength 280nm simultaneously. Maximum field enhancement can reach 20 and 65 fold in the gap of D2PA and CHPA when light incident from substrate, which is expected to greatly enhance fluorescent quantum efficiency that will be confirmed in fluorescence lifetime measurement.

Blue and UV fluorescence of biological fluids and carbon nanodots

NASA Astrophysics Data System (ADS)

Kuznetsov, A.; Frorip, A.; Ots-Rosenberg, M.; Sünter, A.

2013-11-01

Comparative optical study of biofluids (serum, urine, hemodialysate) and carbon nanodots (CND) aqueous solutions has been done. Biofluids were collected from chronic kidney diseases patients (CKD Pts) as well as from normal controls (NCs). Sugar derived CND and oxidized graphene solutions were prepared and used. Fluorescence and excitation spectra have mainly been measured and compared for two sets of subjects. For both family of subjects typical fluorescence with parameters λexсmax/ λemmax = 320+/-5/420+/-5 nm is observed and has many analogeous properties. New effective method of additional similarity identification with use of aluminum salts Al2 (SO4)3, Al (N03)3 and AlCl3 is proposed. Aluminum ions induce the fluorescence band at 380 nm in all substances investigated. Plenty of similar features (12) in optical properties create a united platform for further investigation of the topic - the nature of endogenous near UV and visible fluorescence in biofluids and CND.

Fluorescent bovine serum albumin interacting with the antitussive quencher dextromethorphan: a spectroscopic insight.

PubMed

Durgannavar, Amar K; Patgar, Manjanath B; Nandibewoor, Sharanappa T; Chimatadar, Shivamurti A

2016-05-01

The interaction of dextromethorphan hydrobromide (DXM) with bovine serum albumin (BSA) is studied by using fluorescence spectra, UV-vis absorption, synchronous fluorescence spectra (SFS), 3D fluorescence spectra, Fourier transform infrared (FTIR) spectroscopy and circular dichroism under simulated physiological conditions. DXM effectively quenched the intrinsic fluorescence of BSA. Values of the binding constant, K(A), are 7.159 × 10(3), 9.398 × 10(3) and 16.101 × 10(3)  L/mol; the number of binding sites, n, and the corresponding thermodynamic parameters ΔG°, ΔH° and ΔS° between DXM and BSA were calculated at different temperatures. The interaction between DXM and BSA occurs through dynamic quenching and the effect of DXM on the conformation of BSA was analyzed using SFS. The average binding distance, r, between the donor (BSA) and acceptor (DXM) was determined based on Förster's theory. The results of fluorescence spectra, UV-vis absorption spectra and SFS show that the secondary structure of the protein has been changed in the presence of DXM. Copyright © 2015 John Wiley & Sons, Ltd.

A chemosensor for micro- to nano-molar detection of Ag+ and Hg2+ ions in pure aqueous media and its applications in cell imaging.

PubMed

Nandre, Jitendra P; Patil, Samadhan R; Sahoo, Suban K; Pradeep, Chullikkattil P; Churakov, Andrei; Yu, Fabiao; Chen, Lingxin; Redshaw, Carl; Patil, Ashok A; Patil, Umesh D

2017-10-24

The pyridine substituted thiourea derivative PTB-1 was synthesized and characterized by spectroscopic techniques as well as by single crystal X-ray crystallography. The metal ion sensing ability of PTB-1 was explored by various experimental (naked-eye, UV-Vis, fluorescence, mass spectrometry and 1 H NMR spectroscopy) and theoretical (B3LYP/6-31G**/LANL2DZ) methods. PTB-1 exhibited a highly selective naked-eye detectable color change from colorless to dark brown and UV-Vis spectral changes for the detection of Ag + with a detection limit of 3.67 μM in aqueous medium. The detection of Ag + ions was achieved by test paper strip and supported silica methods. In contrast, PTB-1 exhibited a 23-fold enhanced emission at 420 nm in the presence of Hg 2+ ions with a nano-molar detection limit of 0.69 nM. Finally, the sensor PTB-1 was applied successfully for the intracellular detection of Hg 2+ ions in a HepG2 liver cell line, which was monitored by the use of confocal imaging techniques.

UV/PAR radiations and DOM properties in surface coastal waters of the Canadian shelf of the Beaufort Sea during summer 2009

NASA Astrophysics Data System (ADS)

Para, J.; Charrière, B.; Matsuoka, A.; Miller, W. L.; Rontani, J. F.; Sempéré, R.

2012-11-01

Water masses from the Beaufort Sea in the Arctic Ocean were evaluated for dissolved organic carbon (DOC), and optical characteristics including UV and PAR diffuse attenuation (Kd), and chromophoric and fluorescent dissolved organic matter (CDOM and FDOM) as part of the MALINA field campaign (30 July to 27 August). Even with relatively low mean daily solar radiation incident on the sea surface (0.12 ± 0.03, 8.46 ± 1.64 and 18.09 ± 4.20 kJ m-2 for UV-B (305 nm), UV-A (380 nm) and PAR, respectively), we report significant light penetration with 10% irradiance depths (Z10% (λ)) reaching 9.5 m for 340 nm (UV-A) radiation in the Eastern sector and 4.5 m in the Mackenzie River influenced area (Western sector). Spectral absorption coefficients (aCDOM (350 nm) (m-1)) were significantly correlated to both diffuse attenuation coefficients (Kd) in the UV-A and UV-B and to DOC concentrations. This indicates CDOM as the dominant attenuator of UV solar radiation and suggests its use as an optical proxy for DOC concentrations in this region. Extrapolating CDOM to DOC relationships, we estimate that ~ 16% of the DOC in the Mackenzie River does not absorb radiation at 350 nm. DOC and CDOM discharges by the Mackenzie River during the MALINA Cruise are estimated as ~ 0.22 TgC and 0.18 TgC, respectively. Three dissolved fluorescent components (C1-C3) were identified by fluorescence Excitation/Emission Matrix Spectroscopy (EEMS) and PARAFAC analysis. Our results showed an in-situ biological component (C1) that co-dominated with a terrestrial humic-like component (C2) in the Mackenzie Delta sector, whereas the protein-like (C3) component dominated in the saltiest waters of the North East sector.

Long term storage of virus templated fluorescent materials for sensing applications

NASA Astrophysics Data System (ADS)

Seetharam, Raviraja N.; Szuchmacher Blum, Amy; Soto, Carissa M.; Whitley, Jessica L.; Sapsford, Kim E.; Chatterji, Anju; Lin, Tianwei; Johnson, John E.; Guerra, Charles; Satir, Peter; Ratna, Banahalli R.

2008-03-01

Wild type, mutant, and chemically modified Cowpea mosaic viruses (CPMV) were studied for long term preservation in the presence and absence of cryoprotectants. Viral complexes were reconstituted and tested via fluorescence spectroscopy and a UV/vis-based RNase assay for structural integrity. When viruses lyophilized in the absence of cryoprotectant were rehydrated and RNase treated, UV absorption increased, indicating that the capsids were damaged. The addition of trehalose during lyophilization protected capsid integrity for at least 7 weeks. Measurements of the fluorescence peak maximum of CPMV lyophilized with trehalose and reconstituted also indicate that the virus remained intact. Microarray binding assays indicated that CPMV particles chemically modified for use as a fluorescent tracer were intact and retained binding specificity after lyophilization in the presence of trehalose. Thus, we demonstrate that functionalized CPMV nanostructures can be stored for the long term, enabling their use in practical sensing applications.

Hydrogen bonding-assisted interaction between amitriptyline hydrochloride and hemoglobin: spectroscopic and molecular dynamics studies.

PubMed

Maurya, Neha; Maurya, Jitendra Kumar; Kumari, Meena; Khan, Abbul Bashar; Dohare, Ravins; Patel, Rajan

2017-05-01

Herein, we have explored the interaction between amitriptyline hydrochloride (AMT) and hemoglobin (Hb), using steady-state and time-resolved fluorescence spectroscopy, UV-visible spectroscopy, and circular dichroism spectroscopy, in combination with molecular docking and molecular dynamic (MD) simulation methods. The steady-state fluorescence reveals the static quenching mechanism in the interaction system, which was further confirmed by UV-visible and time-resolved fluorescence spectroscopy. The binding constant, number of binding sites, and thermodynamic parameters viz. ΔG, ΔH, ΔS are also considered; result confirms that the binding of the AMT with Hb is a spontaneous process, involving hydrogen bonding and van der Waals interactions with a single binding site, as also confirmed by molecular docking study. Synchronous fluorescence, CD data, and MD simulation results contribute toward understanding the effect of AMT on Hb to interpret the conformational change in Hb upon binding in aqueous solution.

A Double Decarboxylation in Superfolder Green Fluorescent Protein Leads to High Contrast Photoactivation.

PubMed

Slocum, Joshua D; Webb, Lauren J

2017-07-06

A photoactivatable variant of superfolder green fluorescent protein (GFP) was created by replacing the threonine at position 203 with aspartic acid. Photoactivation by exposure of this mutant to UV light resulted in conversion of the fluorophore from the neutral to the negatively charged form, accompanied by a ∼95-fold increase in fluorescence under 488 nm excitation. Mass spectrometry before and after exposure to UV light revealed a change in mass of 88 Da, attributed to the double decarboxylation of Glu 222 and Asp 203. Kinetics studies and nonlinear power-dependence of the initial rate of photoconversion indicated that the double decarboxylation occurred via a multiphoton absorption process at 254 nm. In addition to providing a photoactivatable GFP with robust folding properties, a detailed mechanistic understanding of this double decarboxylation in GFP will lead to a better understanding of charge transfer in fluorescent proteins.

Status of miniature integrated UV resonance fluorescence and Raman sensors for detection and identification of biochemical warfare agents

NASA Astrophysics Data System (ADS)

Hug, William F.; Bhartia, Rohit; Taspin, Alexandre; Lane, Arthur; Conrad, Pamela; Sijapati, Kripa; Reid, Ray D.

2005-11-01

Laser induced native fluorescence (LINF) is the most sensitive method of detection of biological material including microorganisms, virus', and cellular residues. LINF is also a sensitive method of detection for many non-biological materials as well. The specificity with which these materials can be classified depends on the excitation wavelength and the number and location of observation wavelengths. Higher levels of specificity can be obtained using Raman spectroscopy but a much lower levels of sensitivity. Raman spectroscopy has traditionally been employed in the IR to avoid fluorescence. Fluorescence rarely occurs at wavelength below about 270nm. Therefore, when excitation occurs at a wavelength below 250nm, no fluorescence background occurs within the Raman fingerprint region for biological materials. When excitation occurs within electronic resonance bands of the biological target materials, Raman signal enhancement over one million typically occurs. Raman sensitivity within several hundred times fluorescence are possible in the deep UV where most biological materials have strong absorption. Since the Raman and fluorescence emissions occur at different wavelength, both spectra can be observed simultaneously, thereby providing a sensor with unique sensitivity and specificity capability. We will present data on our integrated, deep ultraviolet, LINF/Raman instruments that are being developed for several applications including life detection on Mars as well as biochemical warfare agents on Earth. We will demonstrate the ability to discriminate organic materials based on LINF alone. Together with UV resonance Raman, higher levels of specificity will be demonstrated. In addition, these instruments are being developed as on-line chemical sensors for industrial and municipal waste streams and product quality applications.

Direct on-strip analysis of size- and time-resolved aerosol impactor samples using laser induced fluorescence spectra excited at 263 and 351 nm.

PubMed

Wang, Chuji; Pan, Yong-Le; James, Deryck; Wetmore, Alan E; Redding, Brandon

2014-04-11

We report a novel atmospheric aerosol characterization technique, in which dual wavelength UV laser induced fluorescence (LIF) spectrometry marries an eight-stage rotating drum impactor (RDI), namely UV-LIF-RDI, to achieve size- and time-resolved analysis of aerosol particles on-strip. The UV-LIF-RDI technique measured LIF spectra via direct laser beam illumination onto the particles that were impacted on a RDI strip with a spatial resolution of 1.2mm, equivalent to an averaged time resolution in the aerosol sampling of 3.6 h. Excited by a 263 nm or 351 nm laser, more than 2000 LIF spectra within a 3-week aerosol collection time period were obtained from the eight individual RDI strips that collected particles in eight different sizes ranging from 0.09 to 10 μm in Djibouti. Based on the known fluorescence database from atmospheric aerosols in the US, the LIF spectra obtained from the Djibouti aerosol samples were found to be dominated by fluorescence clusters 2, 5, and 8 (peaked at 330, 370, and 475 nm) when excited at 263 nm and by fluorescence clusters 1, 2, 5, and 6 (peaked at 390 and 460 nm) when excited at 351 nm. Size- and time-dependent variations of the fluorescence spectra revealed some size and time evolution behavior of organic and biological aerosols from the atmosphere in Djibouti. Moreover, this analytical technique could locate the possible sources and chemical compositions contributing to these fluorescence clusters. Advantages, limitations, and future developments of this new aerosol analysis technique are also discussed. Published by Elsevier B.V.

Apparatus to collect, classify, concentrate, and characterize gas-borne particles

DOEpatents

Rader, Daniel J.; Torczynski, John R.; Wally, Karl; Brockmann, John E.

2003-12-16

An aerosol lab-on-a-chip (ALOC) integrates one or more of a variety of particle collection, classification, concentration (enrichment), an characterization processes onto a single substrate or layered stack of such substrates. By mounting a UV laser diode laser light source on the substrate, or substrates tack, so that it is located down-stream of the sample inlet port and at right angle the sample particle stream, the UV light source can illuminate individual particles in the stream to induce a fluorescence response in those particles having a fluorescent signature such as biological particles, some of said particles. An illuminated particle having a fluorescent signal above a threshold signal would trigger a sorter module that would separate that particle from the particle stream.

Bacterial Sunscreen: Layer-by-Layer Deposition of UV-Absorbing Polymers on Whole-Cell Biosensors (POSTPRINT)

DTIC Science & Technology

2012-06-13

mycosporine - like amino acids that absorb in the UV range and can quench UV-induced intracellular free radicals.2,3 Common in both microorganisms and higher...oxygen, which will react with amino acid side chains and reduce protein stability. GFPuv is excited by long-wave UV and requires ionization for...vinyl sulfate, poly-4-styrenesulfonic acid , and humic acid ) were used to encapsulate E. coli cells expressing green fluorescent protein (GFP) either

Detection of eviscerated poultry spleen enlargement by machine vision

NASA Astrophysics Data System (ADS)

Tao, Yang; Shao, June J.; Skeeles, John K.; Chen, Yud-Ren

1999-01-01

The size of a poultry spleen is an indication of whether the bird is wholesomeness or has a virus-related disease. This study explored the possibility of detecting poultry spleen enlargement with a computer imaging system to assist human inspectors in food safety inspections. Images of 45-day-old hybrid turkey internal viscera were taken using fluorescent and UV lighting systems. Image processing algorithms including linear transformation, morphological operations, and statistical analyses were developed to distinguish the spleen from its surroundings and then to detect abnormal spleens. Experimental results demonstrated that the imaging method could effectively distinguish spleens from other organ and intestine. Based on a total sample of 57 birds, the classification rates were 92% from a self-test set, and 95% from an independent test set for the correct detection of normal and abnormal birds. The methodology indicated the feasibility of using automated machine vision systems in the future to inspect internal organs and check the wholesomeness of poultry carcasses.

New Frontiers in NanoBiotechnology: Monitoring the Protein Function With Single Protein Resolution

DTIC Science & Technology

2005-03-29

Protein (GFP) is a spontaneously fluorescent polypeptide of 27 kD from the jellyfish Aequorea victoria that absorbs UV-blue light and emits in the...will have vast applications in science. Relationship between structure and optical properties in Green Fluorescent Proteins : A quantum mechanical study...RESEARCH AND DEVELOPMENT Invited talks Folding, stability and fluorescence efficiency of the Green and Red Fluorescent Proteins Saverio Alberti Lab.

Dual color microscopic imagery of cells expressing the green fluorescent protein and a red-shifted variant.

PubMed

Yang, T T; Kain, S R; Kitts, P; Kondepudi, A; Yang, M M; Youvan, D C

1996-01-01

The green fluorescent protein (GFP) from the jellyfish, Aequorea victoria, has become a versatile reporter for monitoring gene expression and protein localization in a variety of cells and organisms. GFP emits bright green light (lambda max = 510 nm) when excited with ultraviolet (UV) or blue light (lambda max = 395 nm, minor peak at 470 nm). The chromophore in GFP is intrinsic to the primary structure of the protein, and fluorescence from GFP does not require additional gene products, substrates or other factors. GFP fluorescence is stable, species-independent and can be monitored noninvasively using the techniques of fluorescence microscopy and flow cytometry [Chalfie et al., Science 263 (1994) 802-805; Stearns, Curr. Biol. 5 (1995) 262-264]. The protein appears to undergo an autocatalytic reaction to create the fluorophore [Heim et al., Proc. Natl. Acad. Sci. USA 91 (1994) 12501-12504] in a process involving cyclization of a Tyr66 aa residue. Recently [Delagrave et al., Bio/Technology 13 (1995) 151-154], a combinatorial mutagenic strategy was targeted at aa 64 through 69, which spans the chromophore of A. victoria GFP, yielding a number of different mutants with red-shifted fluorescence excitation spectra. One of these, RSGFP4, retains the characteristic green emission spectra (lambda max = 505 nm), but has a single excitation peak (lambda max = 490 nm). The fluorescence properties of RSGFP4 are similar to those of another naturally occurring GFP from the sea pansy, Renilla reniformis [Ward and Cormier, Photobiochem. Photobiol. 27 (1978) 389-396]. In the present study, we demonstrate by fluorescence microscopy that selective excitation of A. victoria GFP and RSGFP4 allows for spectral separation of each fluorescent signal, and provides the means to image these signals independently in a mixed population of bacteria or mammalian cells.

Integrating fluorescence and interactance measurements to improve apple maturity assessment

NASA Astrophysics Data System (ADS)

Noh, Hyun Kwon; Lu, Renfu

2006-10-01

Fluorescence and reflectance (or interactance) are promising techniques for measuring fruit quality and condition. Our previous research showed that a hyperspectral imaging technique integrating fluorescence and reflectance could improve predictions of selected quality parameters compared to single sensing techniques. The objective of this research was to use a low cost spectrometer for rapid acquisition of fluorescence and interactance spectra from apples and develop an algorithm integrating the two types of data for predicting skin and flesh color, fruit firmness, starch index, soluble solids content, and titratable acid. Experiments were performed to measure UV light induced transient fluorescence and interactance spectra from 'Golden Delicious' apples that were harvested over a period of four weeks during the 2005 harvest season. Standard destructive tests were performed to measure maturity parameters from the apples. Principal component (PC) analysis was applied to the interactance and fluorescence data. A back-propagation feedforward neural network with the inputs of PC data was used to predict individual maturity parameters. Interactance mode was consistently better than fluorescence mode in predicting the maturity parameters. Integrating interactance and fluorescence improved predictions of all parameters except flesh chroma; values of the correlation coefficient for firmness, soluble solids content, starch index, and skin and flesh hue were 0.77, 0.77, 0.89, 0.99, and 0.96 respectively, with the corresponding standard errors of 6.93 N, 0.90%, 0.97 g/L, 0.013 rad, and 0.013 rad. These results represented 4.1% to 23.5% improvements in terms of standard error, in comparison with the better results from the two single sensing methods. Integrating interactance and fluorescence can better assess apple maturity and quality.

Fluorimetric detection of Sn(2+) ion in aqueous medium using Salicylaldehyde based nanoparticles and application to natural samples analysis.

PubMed

Patil, Kishor S; Mahajan, Prasad G; Patil, Shivajirao R

2017-01-05

The fluorescent 2-[(E)-(2-phenylhydrazinylidene)methyl]phenol nanoparticles (PHPNPs) were prepared by a simple reprecipitation method. The prepared PHPNPs examined by Dynamic Light Scattering show narrower particle size distribution having an average particle size of 93.3nm. The Scanning Electron Microphotograph shows distinct spherical shaped morphology of nanoparticles. The blue shift in UV-absorption and fluorescence spectra of PHPNPs with respect to corresponding spectra of PHP in acetone solution indicates H- aggregates and Aggregation Induced Enhanced Emission (AIEE) for nanoparticles. The nanoparticles show selective tendency towards the recognition of Sn(2+) ions by enhancing the fluorescence intensity preference to Cu(2+), Fe(3+), Fe(2+), Ni(2+), NH4(+), Ca(2+), Pb(2+), Hg(2+) and Zn(2+) ions, which actually seem to quench the fluorescence of nanoparticles. The studies on Langmuir adsorption plot, fluorescence lifetime of PHPNPs, DLS-Zeta sizer, UV-visible and fluorescence titration with and without Sn(2+) helped to propose a suitable mechanism of fluorescence enhancement of nanoparticles by Sn(2+) and their binding ability during complexation. The fluorescence enhancement effect of PHPNPs induced by Sn(2+) is further used to develop an analytical method for detection of Sn(2+) from aqueous medium in environmental samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorimetric detection of Sn2 + ion in aqueous medium using Salicylaldehyde based nanoparticles and application to natural samples analysis

NASA Astrophysics Data System (ADS)

Patil, Kishor S.; Mahajan, Prasad G.; Patil, Shivajirao R.

2017-01-01

The fluorescent 2-[(E)-(2-phenylhydrazinylidene)methyl]phenol nanoparticles (PHPNPs) were prepared by a simple reprecipitation method. The prepared PHPNPs examined by Dynamic Light Scattering show narrower particle size distribution having an average particle size of 93.3 nm. The Scanning Electron Microphotograph shows distinct spherical shaped morphology of nanoparticles. The blue shift in UV-absorption and fluorescence spectra of PHPNPs with respect to corresponding spectra of PHP in acetone solution indicates H- aggregates and Aggregation Induced Enhanced Emission (AIEE) for nanoparticles. The nanoparticles show selective tendency towards the recognition of Sn2 + ions by enhancing the fluorescence intensity preference to Cu2 +, Fe3 +, Fe2 +, Ni2 +, NH4+, Ca2 +, Pb2 +, Hg2 + and Zn2 + ions, which actually seem to quench the fluorescence of nanoparticles. The studies on Langmuir adsorption plot, fluorescence lifetime of PHPNPs, DLS-Zeta sizer, UV-visible and fluorescence titration with and without Sn2 + helped to propose a suitable mechanism of fluorescence enhancement of nanoparticles by Sn2 + and their binding ability during complexation. The fluorescence enhancement effect of PHPNPs induced by Sn2 + is further used to develop an analytical method for detection of Sn2 + from aqueous medium in environmental samples.

Detection of Free Thiols and Fluorescence Response of Phycoerythrin Chromophore after Ultraviolet-B Radiation Stress.

PubMed

Kannaujiya, Vinod K; Sinha, Rajeshwar P

2017-03-01

The chemistry of thiol-chromophore linkage plays a central role in the nature of fluorescence of phycoerythrin (PE). Interaction of thiol and chromophore is crucial for the energy transfer, redox signal and inhibition of oxidative damage. In the present investigation the effects of ultraviolet-B radiation on an emission fluorescence intensity and wavelength shift in PE due to interaction between thiol and chromophore by remarkable strategy of detection technique was studied. Purification of PE was done by using a gel permeation and ion exchange chromatography that yielded a quite high purity index (6.40) in a monomeric (αβ) form. UV-B radiation accelerated the quenching efficiency (24.9 ± 1.52%) by reducing fluorescence emission intensity of thiol linked chromophore after 240 min of UV-B exposure. However, after blocking of transiently released free thiol by N-ethylmaleimide, quenching efficiency was increased (36.8 ± 2.80%) with marked emission wavelength shift towards shorter wavelengths up to 562 nm as compared to 575 nm in control. Emission fluorescence of free thiol was at maximum after 240 min that was detected specifically by monobromobimane (mBrB) molecular probe. The association/dissociation of bilin chromophore was analyzed by SDS- and Native-PAGE that also indicated a complete reduction in emission fluorescence. Our work clearly shows an early detection of free thiols and relative interaction with chromophore after UV-B radiation which might play a significant role in structural and functional integrity of terminal PE.

UV-Vis Ratiometric Resonance Synchronous Spectroscopy for Determination of Nanoparticle and Molecular Optical Cross Sections.

PubMed

Nettles, Charles B; Zhou, Yadong; Zou, Shengli; Zhang, Dongmao

2016-03-01

Demonstrated herein is a UV-vis Ratiometric Resonance Synchronous Spectroscopic (R2S2, pronounced as "R-two-S-two" for simplicity) technique where the R2S2 spectrum is obtained by dividing the resonance synchronous spectrum of a NP-containing solution by the solvent resonance synchronous spectrum. Combined with conventional UV-vis measurements, this R2S2 method enables experimental quantification of the absolute optical cross sections for a wide range of molecular and nanoparticle (NP) materials that range optically from pure photon absorbers or scatterers to simultaneous photon absorbers and scatterers, simultaneous photon absorbers and emitters, and all the way to simultaneous photon absorbers, scatterers, and emitters in the UV-vis wavelength region. Example applications of this R2S2 method were demonstrated for quantifying the Rayleigh scattering cross sections of solvents including water and toluene, absorption and resonance light scattering cross sections for plasmonic gold nanoparticles, and absorption, scattering, and on-resonance fluorescence cross sections for semiconductor quantum dots (Qdots). On-resonance fluorescence quantum yields were quantified for the model molecular fluorophore Eosin Y and fluorescent Qdots CdSe and CdSe/ZnS. The insights and methodology presented in this work should be of broad significance in physical and biological science research that involves photon/matter interactions.

Antibiotic Transport in Resistant Bacteria: Synchrotron UV Fluorescence Microscopy to Determine Antibiotic Accumulation with Single Cell Resolution

PubMed Central

Kaščáková, Slávka; Maigre, Laure; Chevalier, Jacqueline; Réfrégiers, Matthieu; Pagès, Jean-Marie

2012-01-01

A molecular definition of the mechanism conferring bacterial multidrug resistance is clinically crucial and today methods for quantitative determination of the uptake of antimicrobial agents with single cell resolution are missing. Using the naturally occurring fluorescence of antibacterial agents after deep ultraviolet (DUV) excitation, we developed a method to non-invasively monitor the quinolones uptake in single bacteria. Our approach is based on a DUV fluorescence microscope coupled to a synchrotron beamline providing tuneable excitation from 200 to 600 nm. A full spectrum was acquired at each pixel of the image, to study the DUV excited fluorescence emitted from quinolones within single bacteria. Measuring spectra allowed us to separate the antibiotic fluorescence from the autofluorescence contribution. By performing spectroscopic analysis, the quantification of the antibiotic signal was possible. To our knowledge, this is the first time that the intracellular accumulation of a clinical antibitiotic could be determined and discussed in relation with the level of drug susceptibility for a multiresistant strain. This method is especially important to follow the behavior of quinolone molecules at individual cell level, to quantify the intracellular concentration of the antibiotic and develop new strategies to combat the dissemination of MDR-bacteria. In addition, this original approach also indicates the heterogeneity of bacterial population when the same strain is under environmental stress like antibiotic attack. PMID:22719907

Light emitting diode excitation emission matrix fluorescence spectroscopy.

PubMed

Hart, Sean J; JiJi, Renée D

2002-12-01

An excitation emission matrix (EEM) fluorescence instrument has been developed using a linear array of light emitting diodes (LED). The wavelengths covered extend from the upper UV through the visible spectrum: 370-640 nm. Using an LED array to excite fluorescence emission at multiple excitation wavelengths is a low-cost alternative to an expensive high power lamp and imaging spectrograph. The LED-EEM system is a departure from other EEM spectroscopy systems in that LEDs often have broad excitation ranges which may overlap with neighboring channels. The LED array can be considered a hybrid between a spectroscopic and sensor system, as the broad LED excitation range produces a partially selective optical measurement. The instrument has been tested and characterized using fluorescent dyes: limits of detection (LOD) for 9,10-bis(phenylethynyl)-anthracene and rhodamine B were in the mid parts-per-trillion range; detection limits for the other compounds were in the low parts-per-billion range (< 5 ppb). The LED-EEMs were analyzed using parallel factor analysis (PARAFAC), which allowed the mathematical resolution of the individual contributions of the mono- and dianion fluorescein tautomers a priori. Correct identification and quantitation of six fluorescent dyes in two to six component mixtures (concentrations between 12.5 and 500 ppb) has been achieved with root mean squared errors of prediction (RMSEP) of less than 4.0 ppb for all components.

Preparation of an Efficient Ratiometric Fluorescent Nanoprobe (m-CDs@[Ru(bpy)3]2+) for Visual and Specific Detection of Hypochlorite on Site and in Living Cells.

PubMed

Zhan, Yuanjin; Luo, Fang; Guo, Longhua; Qiu, Bin; Lin, Yuhong; Li, Juan; Chen, Guonan; Lin, Zhenyu

2017-11-22

Hypochlorite (ClO - ) is one of the most important reactive oxygen species (ROS), which plays an important role in sustaining human innate immunity during microbial invasion. Moreover, ClO - is a powerful oxidizer for water treatment. The safety of drinking water is closely related to its content. Herein, m-phenylenediamine (mPD) is used as a precursor to prepare carbon dots (named m-CDs) with highly fluorescent quantum yield (31.58% in water), and our investigation shows that the strong fluorescent emission of m-CDs can be effectively quenched by ClO - . Based on these findings, we developed a novel fluorescent nanoprobe (m-CDs) for highly selective detection of ClO - . The linear range was from 0.05 to 7 μM (R 2 = 0.998), and the limit of detection (S/N = 3) was as low as 0.012 μM. Moreover, a portable agarose hydrogel solid matrix-based ratiometric fluorescent nanoprobe (m-CDs@[Ru(bpy) 3 ] 2+ ) sensor was subsequently developed for visual on-site detection of ClO - with the naked eyes under a UV lamp, suggesting its potential in practical application with low cost and excellent performance in water quality monitoring. Additionally, intracellular detection of exogenous ClO - was demonstrated via ratiometric imaging microscopy.

Multistimuli-responsive benzothiadiazole-cored phenylene vinylene derivative with nanoassembly properties.

PubMed

Dou, Chuandong; Chen, Dong; Iqbal, Javed; Yuan, Yang; Zhang, Hongyu; Wang, Yue

2011-05-17

A trifluoromethyl-substituted benzothiadiazole-cored phenylene vinylene fluorophore (1) was synthesized and displayed piezo- and vapochromism and thermo-induced fluorescence variation in solid phase. Grinding could disrupt the crystalline compound 1 with orange emission into amorphous compound 1 with green emission, and heating treatment could change the amorphous compound 1 into crystalline compound 1. Ultraviolet-visible (UV-vis) absorption spectra, (13)C nuclear magnetic resonance (NMR), and powder X-ray diffraction (PXRD) characterizations demonstrated that crystalline and amorphous compound 1 possess different molecular packing. A differential scanning calorimetry (DSC) measurement revealed that the emission switching was due to the exchange between the thermodynamic-stable crystalline and metastable amorphous states. The ground sample exhibited vapochromic fluorescence property. Furthermore, compound 1 showed interesting supramolecular assembly characteristics in solution. Slowly cooling the hot N,N-dimethylformamide (DMF) solution of compound 1 resulted in the formation of orange fluorescent fibers, whereas sonication treatment of the cooling solution led to the generation of organic molecular gel. The field emission scanning electronic microscope (FESEM) and fluorescent microscopy images revealed smooth nano- or microfiber and network morphology properties. The PXRD spectra confirmed that these nano- or microstructures had a similar molecular-packing model with the crystalline state of compound 1. Slow evaporation of the toluene solution of compound 1 could produce green emissive microrods, which exhibited interesting thermo-induced fluorescence variation.

Effects of Solar Ultraviolet Radiation on the Potential Efficiency of Photosystem II in Leaves of Tropical Plants1

PubMed Central

Krause, G. Heinrich; Schmude, Claudia; Garden, Hermann; Koroleva, Olga Y.; Winter, Klaus

1999-01-01

The effects of solar ultraviolet (UV)-B and UV-A radiation on the potential efficiency of photosystem II (PSII) in leaves of tropical plants were investigated in Panama (9°N). Shade-grown tree seedlings or detached sun leaves from the outer crown of mature trees were exposed for short periods (up to 75 min) to direct sunlight filtered through plastic or glass filters that absorbed either UV-B or UV-A+B radiation, or transmitted the complete solar spectrum. Persistent changes in potential PSII efficiency were monitored by means of the dark-adapted ratio of variable to maximum chlorophyll a fluorescence. In leaves of shade-grown tree seedlings, exposure to the complete solar spectrum resulted in a strong decrease in potential PSII efficiency, probably involving protein damage. A substantially smaller decline in the dark-adapted ratio of variable to maximum chlorophyll a fluorescence was observed when UV-B irradiation was excluded. The loss in PSII efficiency was further reduced by excluding both UV-B and UV-A light. The photoinactivation of PSII was reversible under shade conditions, but restoration of nearly full activity required at least 10 d. Repeated exposure to direct sunlight induced an increase in the pool size of xanthophyll cycle pigments and in the content of UV-absorbing vacuolar compounds. In sun leaves of mature trees, which contained high levels of UV-absorbing compounds, effects of UV-B on PSII efficiency were observed in several cases and varied with developmental age and acclimation state of the leaves. The results show that natural UV-B and UV-A radiation in the tropics may significantly contribute to photoinhibition of PSII during sun exposure in situ, particularly in shade leaves exposed to full sunlight. PMID:10594122

A compact OPO/SFG laser for ultraviolet biological sensing

NASA Astrophysics Data System (ADS)

Tiihonen, Mikael; Pasiskevicius, Valdas; Laurell, Fredrik; Jonsson, Per; Lindgren, Mikael

2004-07-01

A compact parametric oscillator (OPO) with intracavity sum-frequency generation (SFG) to generate 293 nm UV laser irradiation, was developed. The OPO/SFG device was pumped by a 100 Hz Nd:YAG laser (1064 nm) of own design, including subsequent second harmonic generation (SHG) in an external periodically poled KTiOPO4 (KTP) crystal. The whole system could be used to deliver more than 30 μJ laser irradiation per pulse (100 Hz) at 293 nm. The UV laser light was introduced in an optical fiber attached to a sample compartment allowing detection of fluorescence emission using a commercial spectrometer. Aqueous samples containing biomolecules (ovalbumin) or bacteria spores (Bacillus subtilis) were excited by the UV-light at 293 nm resulting in strong fluorescence emission in the range 325 - 600 nm.

Oxygen Sensing Based on the Yellowing of Newspaper.

PubMed

Yu, Jingjing; Qin, Xingcai; Xian, Xiaojun; Tao, Nongjian

2018-01-26

Newspaper is known to turn yellow over time. We show here that this yellowing process is sensitive to oxygen when exposed to UV light, leading to oxygen sensing. Oxygen sensing is critical to many applications, including industrial process control and breath analysis, but the existing oxygen sensors have limitations, especially for breath analysis that operates at 100% humidity. The UV irradiation also triggers fluorescence emission from newspaper, and the fluorescence intensity depends on oxygen concentration, providing an additional oxygen sensing method. Newspaper is stable in ambient air, and reactive to oxygen only with UV activation, which overcomes the instability issue of a typical colorimetric sensor in ambient air. The newspaper oxygen sensor works in 100% relative humidity air, containing various interferents. These unique properties of newspaper promise low cost and reliable oxygen sensing applications.

Fluorescence Fluctuations and Equivalence Classes of Ca 2+ Imaging Experiments

PubMed Central

Piegari, Estefanía; Lopez, Lucía; Perez Ipiña, Emiliano; Ponce Dawson, Silvina

2014-01-01

release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP3Rs) plays a relevant role in numerous physiological processes. IP3R-mediated signals involve -induced -release (CICR) whereby release through one open IP3R induces the opening of other channels. IP3Rs are apparently organized in clusters. The signals can remain localized (i.e., puffs) if CICR is limited to one cluster or become waves that propagate between clusters. puffs are the building blocks of waves. Thus, there is great interest in determining puff properties, especially in view of the current controversy on the spatial distribution of activatable IP3Rs. puffs have been observed in intact cells with optical techniques proving that they are intrinsically stochastic. Obtaining a correct picture of their dynamics then entails being able to detect the whole range of puff sizes. puffs are observed using visible single-wavelength dyes, slow exogenous buffers (e.g., EGTA) to disrupt inter-cluster CICR and UV-photolyzable caged IP3. Single-wavelength dyes increase their fluorescence upon calcium binding producing images that are strongly dependent on their kinetic, transport and photophysical properties. Determining the artifacts that the imaging setting introduces is particularly relevant when trying to analyze the smallest signals. In this paper we introduce a method to estimate the expected signal-to-noise ratio of imaging experiments that use single-wavelength dyes. The method is based on the Number and Brightness technique. It involves the performance of a series of experiments and their subsequent analysis in terms of a fluorescence fluctuation model with which the model parameters are quantified. Using the model, the expected signal-to-noise ratio is then computed. Equivalence classes between different experimental conditions that produce images with similar signal-to-noise ratios can then be established. The method may also be used to estimate the smallest signals that can reliably be observed with each setting. PMID:24776736

Visual outdoor response of multiple wild bee species: highly selective stimulation of a single photoreceptor type by sunlight-induced fluorescence.

PubMed

Rao, Sujaya; Ostroverkhova, Oksana

2015-07-01

Bees have ultraviolet (UV), blue and green photoreceptor types in their compound eyes with which they locate food sources in landscapes that change continuously in cues emanating from plants and backgrounds against which they are perceived. The complexity of bee vision has been elucidated through studies examining individual species under laboratory conditions. Here, we used a bee-attractive fluorescent blue trap as a model for analyzing visual signals in operation outdoors, and across bee species. We manipulated trap color (appearance to humans under light with weak UV component) and UV-induced fluorescence emission, and aligned field capture results with bee vision models. Our studies show that the bees were attracted to traps that under solar illumination exhibited strong fluorescence emission exclusively in the blue spectral region. Through quantitative analysis, we established that strong spectral overlap of trap emittance with the photosensitivity characteristic of the blue receptor type and minimal overlap with those of the other two receptor types is the most critical property of attractive traps. A parameter has been identified which predicts the degree of attractiveness of the traps and which captures trends in the field data across wild bee species and for a diversity of backgrounds.

UV-Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 2: selected applications.

PubMed

Antosiewicz, Jan M; Shugar, David

In Part 2 we discuss application of several different types of UV-Vis spectroscopy, such as normal, difference, and second-derivative UV absorption spectroscopy, fluorescence spectroscopy, linear and circular dichroism spectroscopy, and Raman spectroscopy, of the side-chain of tyrosine residues in different molecular environments. We review the ways these spectroscopies can be used to probe complex protein structures.

UV-Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 2: selected applications.

PubMed

Antosiewicz, Jan M; Shugar, David

2016-06-01

In Part 2 we discuss application of several different types of UV-Vis spectroscopy, such as normal, difference, and second-derivative UV absorption spectroscopy, fluorescence spectroscopy, linear and circular dichroism spectroscopy, and Raman spectroscopy, of the side-chain of tyrosine residues in different molecular environments. We review the ways these spectroscopies can be used to probe complex protein structures.

Detection of Organic Matter in Greenland Ice Cores by Deep-UV Fluorescence

NASA Astrophysics Data System (ADS)

Willis, M.; Malaska, M.; Wanger, G.; Bhartia, R.; Eshelman, E.; Abbey, W.; Priscu, J. C.

2017-12-01

The Greenland Ice Sheet is an Earthly analog for icy ocean worlds in the outer Solar System. Future missions to such worlds including Europa, Enceladus, and Titan may potentially include spectroscopic instrumentation to examine the surface/subsurface. The primary goal of our research is to test deep UV/Raman systems for in the situ detection and localization of organics in ice. As part of this effort we used a deep-UV fluorescence instrument able to detect naturally fluorescent biological materials such as aromatic molecules found in proteins and whole cells. We correlated these data with more traditional downstream analyses of organic material in natural ices. Supraglacial ice cores (2-4 m) were collected from several sites on the southwest outlet of the Greenland Ice Sheet using a 14-cm fluid-free mechanical coring system. Repeat spectral mapping data were initially collected longitudinally on uncut core sections. Cores were then cut into 2 cm thick sections along the longitudinal axis, slowly melted and analyzed for total organic carbon (TOC), total dissolved nitrogen (TDN), and bacterial density. These data reveal a spatial correlation between organic matter concentration, cell density, and the deep UV fluorescence maps. Our results provide a profile of the organics embedded within the ice from the top surface into the glacial subsurface, and the TOC:TDN data from the clean interior of the cores are indicative of a biological origin. This work provides a background dataset for future work to characterize organic carbon in the Greenland Ice Sheet and validation of novel instrumentation for in situ data collection on icy bodies.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, David C.; Kempe, Michael D.; Muller, Matthew T.

We examined the durability of polymeric encapsulation materials using outdoor exposure at the nominal geometric concentration of 500 suns. The results for 36-month cumulative field deployment are presented for materials including: poly(ethylene-co-vinyl acetate), (EVA); polyvinyl butyral (PVB); ionomer; polyethylene/polyoctene copolymer (PO); thermoplastic polyurethane (TPU); poly(dimethylsiloxane) (PDMS); poly(diphenyl dimethyl siloxane) (PDPDMS); and poly(phenyl-methyl siloxane) (PPMS). Measurements of the field conditions including ambient temperature and ultraviolet (UV) dose were recorded at the test site during the experiment. Our measurements for the experiment included optical transmittance (with subsequent analysis of solar-weighted transmittance, UV cut-off wavelength, and yellowness index), mass, visual photography, photoelastic imaging,more » and fluorescence spectroscopy. While the results to date for EVA are presented and discussed, examination here focuses more on the siloxane materials. A specimen recently observed to fail by thermal decomposition is discussed in terms of the implementation of the experiment as well as its fluorescence signature, which was observed to become more pronounced with age. Modulated thermogravimetry (allowing determination of the activation energy of thermal decomposition) was performed on a subset of the siloxanes to quantify the propensity for decomposition at elevated temperatures. Supplemental, Pt-catalyst- and primer-solutions as well as peroxide-cured PDMS specimens were examined to assess the source of the luminescence. Furthermore, our results, including the change in optical transmittance, observed failure modes, and subsequent analyses of the failure modes are described in the conclusions.« less

Novel tissue phantom for testing a dual-modality diagnostic system: time-resolved fluorescence spectroscopy and high frequency ultrasound

NASA Astrophysics Data System (ADS)

Sun, Yang; Liao, Kuo-Chih; Sun, Yinghua; Park, Jesung; Marcu, Laura

2008-02-01

A unique tissue phantom is reported here that mimics the optical and acoustical properties of biological tissue and enables testing and validation of a dual-modality clinical diagnostic system combining time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) and ultrasound backscatter microscopy (UBM). The phantom consisted of contrast agents including silicon dioxide particles with a range of diameters from 0.5 to 10 μm acting as optical and acoustical scatterers, and FITC-conjugated dextran mimicking the endogenous fluorophore in tissue. The agents were encapsulated in a polymer bead attached to the end of an optical fiber with a 200 μm diameter using a UV-induced polymerization technique. A set of beads with fibers were then implanted into a gel-based matrix with controlled patterns including a design with lateral distribution and a design with successively changing depth. The configuration presented here allowed the validation of the hybrid fluorescence spectroscopic and ultrasonic system by detecting the lateral and depth distribution of the contrast agents, as well as for coregistration of the ultrasonic image with spectroscopic data. In addition, the depth of the beads in the gel matrix was changed to explore the effect of different concentration ratio of the mixture on the fluorescence signal emitted.

Fluorescence development of fingerprints by combining conjugated polymer nanoparticles with cyanoacrylate fuming.

PubMed

Chen, Hong; Ma, Rong-Liang; Fan, Zhinan; Chen, Yun; Wang, Zizheng; Fan, Li-Juan

2018-05-23

Selecting appropriate developing methods/reagents or their combination to enhance the effect for fingerprint development is of great significance for practical forensic investigation. Ethyl-2-cyanoacrylate ester (superglue) fuming is a popular method for "in-situ" developing fingerprints in forensic science, followed by fluorescence staining to enhance the contrast of the fingerprint image in some occasion. In this study, a series of fluorescent poly(p-phenylene vinylene) (PPV) nanoparticles (NPs) in colloidal solution were successfully prepared and the emission color was tuned via a simple way. The fuming process was carried out using a home-made device. The staining was accomplished by immersing a piece of absorbent cotton into the solution of NPs, and then gently applied on the fumed fingerprints for several times. The PPV NPs were found to have a better developing effect than Rhodamine 6G when excited by 365 nm UV lamp. Different emission colors of NPs are advantageous in developing fingerprints on various substrates. Mechanism study suggested that the NPs were embedded in the porous structure of the superglue resin. In all, the combination of fuming method with the staining by conjugated polymer NPs has been demonstrated to be successful for fluorescent fingerprint development and be promising for more practical forensic applications. Copyright © 2018. Published by Elsevier Inc.

Enhancement of cell internalization and photostability of red and green emitter quantum dots upon entrapment in novel cationic nanoliposomes.

PubMed

Samadikhah, Hamid Reza; Nikkhah, Maryam; Hosseinkhani, Saman

2017-06-01

Two quantum dots (QDs), a green emitter, CdSe and a red emitter, CdSe with ZnS shell are encapsulated into novel liposomes in two different formulations including cationic liposomes. Quantum dots have proven themselves as powerful inorganic fluorescent probes, especially for long-term, multiplexed imaging and detection. Upon delivery into a cell, in endocytic vesicles such as endosomes, their fluorescence is quenched. We have investigated the potential toxic effects, photophysical properties and cell internalization of QDs in new formulation of liposomes as an in vitro vesicle model. Entrapment of QDs into liposomes is brought about with a decrease in their intrinsic fluorescence and toxicities and an increase in their photostability and lifetime. The biomimetic lipid bilayer of liposomes provides high biocompatibility, thereby enhancing the effectiveness of fluorescent nanoparticles for biological recognition in vitro and in vivo. The prepared lipodots could effectively prevent QDs from photo-oxidation during storage and when exposed to ultraviolet (UV) light. Moreover, the flow cytometry of HEK 293 T cells showed that the cell internalization of encapsulated QDs in (DSPC/CHO/DOPE/DOAB) liposome is enhanced 10 times compared with non-encapsulated QD (bare QDs). Copyright © 2016 John Wiley & Sons, Ltd.

Simple, fast, and low-cost camera-based water content measurement with colorimetric fluorescent indicator

NASA Astrophysics Data System (ADS)

Song, Seok-Jeong; Kim, Tae-Il; Kim, Youngmi; Nam, Hyoungsik

2018-05-01

Recently, a simple, sensitive, and low-cost fluorescent indicator has been proposed to determine water contents in organic solvents, drugs, and foodstuffs. The change of water content leads to the change of the indicator's fluorescence color under the ultra-violet (UV) light. Whereas the water content values could be estimated from the spectrum obtained by a bulky and expensive spectrometer in the previous research, this paper demonstrates a simple and low-cost camera-based water content measurement scheme with the same fluorescent water indicator. Water content is calculated over the range of 0-30% by quadratic polynomial regression models with color information extracted from the captured images of samples. Especially, several color spaces such as RGB, xyY, L∗a∗b∗, u‧v‧, HSV, and YCBCR have been investigated to establish the optimal color information features over both linear and nonlinear RGB data given by a camera before and after gamma correction. In the end, a 2nd order polynomial regression model along with HSV in a linear domain achieves the minimum mean square error of 1.06% for a 3-fold cross validation method. Additionally, the resultant water content estimation model is implemented and evaluated in an off-the-shelf Android-based smartphone.

Systematic characterization and fluorescence threshold strategies for the wideband integrated bioaerosol sensor (WIBS) using size-resolved biological and interfering particles

NASA Astrophysics Data System (ADS)

Savage, Nicole J.; Krentz, Christine E.; Könemann, Tobias; Han, Taewon T.; Mainelis, Gediminas; Pöhlker, Christopher; Huffman, J. Alex

2017-11-01

Atmospheric particles of biological origin, also referred to as bioaerosols or primary biological aerosol particles (PBAP), are important to various human health and environmental systems. There has been a recent steep increase in the frequency of published studies utilizing commercial instrumentation based on ultraviolet laser/light-induced fluorescence (UV-LIF), such as the WIBS (wideband integrated bioaerosol sensor) or UV-APS (ultraviolet aerodynamic particle sizer), for bioaerosol detection both outdoors and in the built environment. Significant work over several decades supported the development of the general technologies, but efforts to systematically characterize the operation of new commercial sensors have remained lacking. Specifically, there have been gaps in the understanding of how different classes of biological and non-biological particles can influence the detection ability of LIF instrumentation. Here we present a systematic characterization of the WIBS-4A instrument using 69 types of aerosol materials, including a representative list of pollen, fungal spores, and bacteria as well as the most important groups of non-biological materials reported to exhibit interfering fluorescent properties. Broad separation can be seen between the biological and non-biological particles directly using the five WIBS output parameters and by taking advantage of the particle classification analysis introduced by Perring et al. (2015). We highlight the importance that particle size plays on observed fluorescence properties and thus in the Perring-style particle classification. We also discuss several particle analysis strategies, including the commonly used fluorescence threshold defined as the mean instrument background (forced trigger; FT) plus 3 standard deviations (σ) of the measurement. Changing the particle fluorescence threshold was shown to have a significant impact on fluorescence fraction and particle type classification. We conclude that raising the fluorescence threshold from FT + 3σ to FT + 9σ does little to reduce the relative fraction of biological material considered fluorescent but can significantly reduce the interference from mineral dust and other non-biological aerosols. We discuss examples of highly fluorescent interfering particles, such as brown carbon, diesel soot, and cotton fibers, and how these may impact WIBS analysis and data interpretation in various indoor and outdoor environments. The performance of the particle asymmetry factor (AF) reported by the instrument was assessed across particle types as a function of particle size, and comments on the reliability of this parameter are given. A comprehensive online supplement is provided, which includes size distributions broken down by fluorescent particle type for all 69 aerosol materials and comparing threshold strategies. Lastly, the study was designed to propose analysis strategies that may be useful to the broader community of UV-LIF instrumentation users in order to promote deeper discussions about how best to continue improving UV-LIF instrumentation and results.

Analysis of conjugation of chloramphenicol and hemoglobin by fluorescence, circular dichroism and molecular modeling

NASA Astrophysics Data System (ADS)

Ding, Fei; Liu, Wei; Sun, Ye; Yang, Xin-Ling; Sun, Ying; Zhang, Li

2012-01-01

Chloramphenicol is a low cost, broad spectrum, highly active antibiotic, and widely used in the treatment of serious infections, including typhoid fever and other life-threatening infections of the central nervous system and respiratory tract. The purpose of the present study was to examine the conjugation of chloramphenicol with hemoglobin (Hb) and compared with albumin at molecular level, utilizing fluorescence, UV/vis absorption, circular dichroism (CD) as well as molecular modeling. Fluorescence data indicate that drug bind Hb generate quenching via static mechanism, this corroborates UV/vis absorption measurements that the ground state complex formation with an affinity of 10 4 M -1, and the driving forces in the Hb-drug complex are hydrophilic interactions and hydrogen bonds, as derived from computational model. The accurate binding site of drug has been identified from the analysis of fluorescence and molecular modeling, α1β2 interface of Hb was assigned to possess high-affinity for drug, which located at the β-37 Trp nearby. The structural investigation of the complexed Hb by synchronous fluorescence, UV/vis absorption, and CD observations revealed some degree of Hb structure unfolding upon complexation. Based on molecular modeling, we can draw the conclusion that the binding affinity of drug with albumin is superior, compared with Hb. These phenomena can provide salient information on the absorption, distribution, pharmacology, and toxicity of chloramphenicol and other drugs which have analogous configuration with chloramphenicol.

Using resonance light scattering and UV/vis absorption spectroscopy to study the interaction between gliclazide and bovine serum albumin.

PubMed

Zhang, Qiu-Ju; Liu, Bao-Sheng; Li, Gai-Xia; Han, Rong

2016-08-01

At different temperatures (298, 310 and 318 K), the interaction between gliclazide and bovine serum albumin (BSA) was investigated using fluorescence quenching spectroscopy, resonance light scattering spectroscopy and UV/vis absorption spectroscopy. The first method studied changes in the fluorescence of BSA on addition of gliclazide, and the latter two methods studied the spectral change in gliclazide while BSA was being added. The results indicated that the quenching mechanism between BSA and gliclazide was static. The binding constant (Ka ), number of binding sites (n), thermodynamic parameters, binding forces and Hill's coefficient were calculated at three temperatures. Values for the binding constant obtained using resonance light scattering and UV/vis absorption spectroscopy were much greater than those obtained from fluorescence quenching spectroscopy, indicating that methods monitoring gliclazide were more accurate and reasonable. In addition, the results suggest that other residues are involved in the reaction and the mode 'point to surface' existed in the interaction between BSA and gliclazide. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

SHERLOC on Mars 2020

NASA Astrophysics Data System (ADS)

Beegle, L. W.; Bhartia, R.; DeFlores, L. P.; Abbey, W.; Asher, S. A.; Burton, A. S.; Fries, M.; Conrad, P. G.; Clegg, S. M.; Wiens, R. C.; Edgett, K. S.; Ehlmann, B. L.; Nealson, K. H.; Minitti, M. E.; Popp, J.; Langenhorst, F.; Sobron, P.; Steele, A.; Williford, K. H.; Yingst, R. A.

2017-12-01

The Scanning Habitable Environments with Raman & Luminescence for Organics & Chemicals (SHERLOC) investigation is part of the Mars 2020 integrated payload. SHERLOC enables non-contact, spatially resolved, and highly sensitivity detection and characterization of organics and minerals in the Martian surface and near subsurface. SHERLOC is an arm-mounted, Deep UV (DUV) resonance Raman and fluorescence spectrometer utilizing a 248.6-nm DUV laser. Deep UV induced native fluorescence is very sensitive to condensed carbon and aromatic organics, enabling detection at or below 10-6 w/w (1 ppm) at <100 µm spatial scales. SHERLOC's deep UV resonance Raman enables detection and classification of aromatic and aliphatic organics with sensitivities of 10-2 to below 10-4 w/w. In addition to organics, the deep UV Raman enables detection and classification of minerals relevant to aqueous chemistry with grain sizes below 20 µm. SHERLOC will be able to map the distribution of organic material with respect to visible features and minerals that are identifiable with the Raman spectrometer. These maps will enable analysis of the distribution of organics with minerals.

Photosynthetic benefits of ultraviolet-A to Pimelea ligustrina, a woody shrub of sub-alpine Australia.

PubMed

Turnbull, Tarryn L; Barlow, Alexandra M; Adams, Mark A

2013-10-01

The definition of photosynthetically active radiation (Q) as the visible waveband (λ 400-700 nm) is a core assumption of much of modern plant biology and global models of carbon and water fluxes. On the other hand, much research has focused on potential mutation and damage to leaves caused by ultraviolet (UV) radiation (280-400 nm), and anatomical and physiological adaptations that help avoid such damage. Even so, plant responses to UV-A are poorly described and, until now, photosynthetic utilization of UV-A has not been elucidated under full light conditions in the field. We found that the UV-A content of sunlight increased photosynthetic rates in situ by 12% in Pimelea ligustrina Labill., a common and indigenous woody shrub of alpine ecosystems of the Southern Hemisphere. Compared to companion shrubs, UV-A-induced photosynthesis in P. ligustrina resulted from reduced physical and chemical capacities to screen UV-A at the leaf surface (illustrated by a lack of cuticle and reduced phenol index) and the resulting ability of UV-A to excite chlorophyll (Chl) a directly, and via energy provided by the carotenoid lutein. A screening of 55 additional sub-alpine species showed that 47% of the plant taxa also display Chl a fluorescence under UV-A. If Chl a fluorescence indicates potential for photosynthetic gain, continued exclusion of UV-A from definitions of Q in this ecosystem could result in underestimates of measured and modeled rates of photosynthesis and miscalculation of potential for carbon sequestration. We suggest that carbon gain for alpine environs across the globe could be similarly underestimated given that UV-A radiation increases with altitude and that the frequently dominant herb and grass life-forms often transmit UV-A through the epidermis.

Ultraviolet light emitting diodes and bio-aerosol sensing

NASA Astrophysics Data System (ADS)

Davitt, Kristina M.

Recent interest in compact ultraviolet (UV) light emitters has produced advances in material quality and device performance from aluminum-rich alloys of the nitride semiconductor system. The epitaxial growth of device structures from this material poses remarkable challenges, and state-of-the-art in semiconductor UV light sources at wavelengths shorter than 350 nm is currently limited to LEDs. A portion of the work presented in this thesis involves the design and characterization of UV LED structures, with particular focus on sub-300 nm LEDs which have only been demonstrated within the last four years. Emphasis has been placed on the integration of early devices with modest efficiencies and output powers into a practical, fluorescence-based bio-sensing instrument. The quality of AlGaInN and AlGaN-based materials is characterized by way of the performance of 340 nm and 290 nm LEDs respectively. A competitive level of device operation is achieved, although much room remains for improvement in the efficiency of light emission from this material system. A preliminary investigation of 300 nm LEDs grown on bulk AIN shows promising electrical and optical characteristics, and illustrates the numerous advantages that this native substrate offers to the epitaxy of wide bandgap nitride semiconductors. The application of UV LEDs to the field of bio-aerosol sensing is pursued by constructing an on-the-fly fluorescence detection system. A linear array of UV LEDs is designed and implemented, and the capability of test devices to excite native fluorescence from bacterial spores is established. In order to fully capitalize on the reduction in size afforded by LEDs, effort is invested in re-engineering the remaining sensor components. Operation of a prototype system for physically sorting bio-aerosols based on fluorescence spectra acquired in real-time from single airborne particles excited by a UV-LED array is demonstrated using the bio-fluorophores NADH and tryptophan. Sensor performance is shown to be ultimately linked to the material quality of high aluminum fraction nitrides, and is expected to show progress as this field matures.

Observations of fluorescent aerosol-cloud interactions in the free troposphere at the Sphinx high Alpine research station, Jungfraujoch

NASA Astrophysics Data System (ADS)

Crawford, I.; Lloyd, G.; Bower, K. N.; Connolly, P. J.; Flynn, M. J.; Kaye, P. H.; Choularton, T. W.; Gallagher, M. W.

2015-09-01

The fluorescent nature of aerosol at a high Alpine site was studied using a wide-band integrated bioaerosol (WIBS-4) single particle multi-channel ultra violet-light induced fluorescence (UV-LIF) spectrometer. This was supported by comprehensive cloud microphysics and meteorological measurements with the aims of cataloguing concentrations of bio-fluorescent aerosols at this high altitude site and also investigating possible influences of UV-fluorescent particle types on cloud-aerosol processes. Analysis of background free tropospheric air masses, using a total aerosol inlet, showed there to be a minor but statistically insignificant increase in the fluorescent aerosol fraction during in-cloud cases compared to out of cloud cases. The size dependence of the fluorescent aerosol fraction showed the larger aerosol to be more likely to be fluorescent with 80 % of 10 μm particles being fluorescent. Whilst the fluorescent particles were in the minority (NFl/NAll = 0.27±0.19), a new hierarchical agglomerative cluster analysis approach, Crawford et al. (2015) revealed the majority of the fluorescent aerosol were likely to be representative of fluorescent mineral dust. A minor episodic contribution from a cluster likely to be representative of primary biological aerosol particles (PBAP) was also observed with a wintertime baseline concentration of 0.1±0.4 L-1. Given the low concentration of this cluster and the typically low ice active fraction of studied PBAP (e.g. pseudomonas syringae) we suggest that the contribution to the observed ice crystal concentration at this location is not significant during the wintertime.

Widespread bone-based fluorescence in chameleons.

PubMed

Prötzel, David; Heß, Martin; Scherz, Mark D; Schwager, Martina; Padje, Anouk Van't; Glaw, Frank

2018-01-15

Fluorescence is widespread in marine organisms but uncommon in terrestrial tetrapods. We here show that many chameleon species have bony tubercles protruding from the skull that are visible through their scales, and fluoresce under UV light. Tubercles arising from bones of the skull displace all dermal layers other than a thin, transparent layer of epidermis, creating a 'window' onto the bone. In the genus Calumma, the number of these tubercles is sexually dimorphic in most species, suggesting a signalling role, and also strongly reflects species groups, indicating systematic value of these features. Co-option of the known fluorescent properties of bone has never before been shown, yet it is widespread in the chameleons of Madagascar and some African chameleon genera, particularly in those genera living in forested, humid habitats known to have a higher relative component of ambient UV light. The fluorescence emits with a maximum at around 430 nm in blue colour which contrasts well to the green and brown background reflectance of forest habitats. This discovery opens new avenues in the study of signalling among chameleons and sexual selection factors driving ornamentation.

QSPR studies on the photoinduced-fluorescence behaviour of pharmaceuticals and pesticides.

PubMed

López-Malo, D; Bueso-Bordils, J I; Duart, M J; Alemán-López, P A; Martín-Algarra, R V; Antón-Fos, G M; Lahuerta-Zamora, L; Martínez-Calatayud, J

2017-07-01

Fluorimetric analysis is still a growing line of research in the determination of a wide range of organic compounds, including pharmaceuticals and pesticides, which makes necessary the development of new strategies aimed at improving the performance of fluorescence determinations as well as the sensitivity and, especially, the selectivity of the newly developed analytical methods. In this paper are presented applications of a useful and growing tool suitable for fostering and improving research in the analytical field. Experimental screening, molecular connectivity and discriminant analysis are applied to organic compounds to predict their fluorescent behaviour after their photodegradation by UV irradiation in a continuous flow manifold (multicommutation flow assembly). The screening was based on online fluorimetric measurement and comprised pre-selected compounds with different molecular structures (pharmaceuticals and some pesticides with known 'native' fluorescent behaviour) to study their changes in fluorescent behaviour after UV irradiation. Theoretical predictions agree with the results from the experimental screening and could be used to develop selective analytical methods, as well as helping to reduce the need for expensive, time-consuming and trial-and-error screening procedures.

Characterization of low-dose doxorubicin-loaded silica-based nanocomposites

NASA Astrophysics Data System (ADS)

Prokopowicz, Magdalena

2018-01-01

In this study, we synthesized multicomponent solid films of low-dose doxorubicin (DOX)-loaded polydimethylsiloxane (PDMS)-SiO2/CaP nanocomposites via sol-gel process combined with the method of evaporation-induced self-assembly (EISA) at low temperature. Nanomechanical properties (elasticity and adhesion) of the synthesized multicomponent films were determined by using atomic force microscopy with a PeakForce™ quantitative nanomechanical mapping imaging technique. Solid state of DOX in the synthesized films was studied by using UV-vis and fluorescence spectroscopy. The release profile of different concentrations of DOX loaded (1, 3, and 5 wt%) on the multicomponent films was assessed using USP Apparatus 4 and via UV-vis end analysis. Results indicate drug-component interactions on the overall morphology of domains (size and shape), nanomechanical properties, and release behavior of the DOX-loaded nanocomposites. We observed a progressive increase in surface roughness and mean adhesive value with increasing concentration of DOX loaded (0-5 wt%). In addition, for all the different concentrations of DOX-loaded, we observed a diffusion-controlled drug release.

A look into the invisible: ultraviolet-B sensitivity in an insect (Caliothrips phaseoli) revealed through a behavioural action spectrum

PubMed Central

Mazza, Carlos A.; Izaguirre, Miriam M.; Curiale, Javier; Ballaré, Carlos L.

2010-01-01

Caliothrips phaseoli, a phytophagous insect, detects and responds to solar ultraviolet-B radiation (UV-B; λ ≤ 315 nm) under field conditions. A highly specific mechanism must be present in the thrips visual system in order to detect this narrow band of solar radiation, which is at least 30 times less abundant than the UV-A (315–400 nm), to which many insects are sensitive. We constructed an action spectrum of thrips responses to light by studying their behavioural reactions to monochromatic irradiation under confinement conditions. Thrips were maximally sensitive to wavelengths between 290 and 330 nm; human-visible wavelengths (λ ≥ 400 nm) failed to elicit any response. All but six ommatidia of the thrips compound eye were highly fluorescent when exposed to UV-A of wavelengths longer than 330 nm. We hypothesized that the fluorescent compound acts as an internal filter, preventing radiation with λ > 330 nm from reaching the photoreceptor cells. Calculations based on the putative filter transmittance and a visual pigment template of λmax = 360 nm produced a sensitivity spectrum that was strikingly similar to the action spectrum of UV-induced behavioural response. These results suggest that specific UV-B vision in thrips is achieved by a standard UV-A photoreceptor and a sharp cut-off internal filter that blocks longer UV wavelengths in the majority of the ommatidia. PMID:19846453

The research on a novel type of the solar-blind UV head-mounted displays

NASA Astrophysics Data System (ADS)

Zhao, Shun-long

2011-08-01

Ultraviolet technology of detecting is playing a more and more important role in the field of civil application, especially in the corona discharge detection, in modern society. Now the UV imaging detector is one of the most important equipments in power equipment flaws detection. And the modern head-mounted displays (HMDs) have shown the applications in the fields of military, industry production, medical treatment, entertainment, 3D visualization, education and training. We applied the system of head-mounted displays to the UV image detection, and a novel type of head-mounted displays is presented: the solar-blind UV head-mounted displays. And the structure is given. By the solar-blind UV head-mounted displays, a real-time, isometric and visible image of the corona discharge is correctly displayed upon the background scene where it exists. The user will see the visible image of the corona discharge on the real scene rather than on a small screen. Then the user can easily find out the power equipment flaws and repair them. Compared with the traditional UV imaging detector, the introducing of the HMDs simplifies the structure of the whole system. The original visible spectrum optical system is replaced by the eye in the solar-blind UV head-mounted displays. And the optical image fusion technology would be used rather than the digital image fusion system which is necessary in traditional UV imaging detector. That means the visible spectrum optical system and digital image fusion system are not necessary. This makes the whole system cheaper than the traditional UV imaging detector. Another advantage of the solar-blind UV head-mounted displays is that the two hands of user will be free. So while observing the corona discharge the user can do some things about it. Therefore the solar-blind UV head-mounted displays can make the corona discharge expose itself to the user in a better way, and it will play an important role in corona detection in the future.

UV-Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 1: basic principles and properties of tyrosine chromophore.

PubMed

Antosiewicz, Jan M; Shugar, David

Spectroscopic properties of tyrosine residues may be employed in structural studies of proteins. Here we discuss several different types of UV-Vis spectroscopy, like normal, difference and second-derivative UV absorption spectroscopy, fluorescence spectroscopy, linear and circular dichroism spectroscopy, and Raman spectroscopy, and corresponding optical properties of the tyrosine chromophore, phenol, which are used to study protein structure.

UV-Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 1: basic principles and properties of tyrosine chromophore.

PubMed

Antosiewicz, Jan M; Shugar, David

2016-06-01

Spectroscopic properties of tyrosine residues may be employed in structural studies of proteins. Here we discuss several different types of UV-Vis spectroscopy, like normal, difference and second-derivative UV absorption spectroscopy, fluorescence spectroscopy, linear and circular dichroism spectroscopy, and Raman spectroscopy, and corresponding optical properties of the tyrosine chromophore, phenol, which are used to study protein structure.

Dual-Targeting Lactoferrin-Conjugated Polymerized Magnetic Polydiacetylene-Assembled Nanocarriers with Self-Responsive Fluorescence/Magnetic Resonance Imaging for In Vivo Brain Tumor Therapy.

PubMed

Fang, Jen-Hung; Chiu, Tsung-Lang; Huang, Wei-Chen; Lai, Yen-Ho; Hu, Shang-Hsiu; Chen, You-Yin; Chen, San-Yuan

2016-03-01

Maintaining a high concentration of therapeutic agents in the brain is difficult due to the restrictions of the blood-brain barrier (BBB) and rapid removal from blood circulation. To enable controlled drug release and enhance the blood-brain barrier (BBB)-crossing efficiency for brain tumor therapy, a new dual-targeting magnetic polydiacetylene nanocarriers (PDNCs) delivery system modified with lactoferrin (Lf) is developed. The PDNCs are synthesized using the ultraviolet (UV) cross-linkable 10,12-pentacosadiynoic acid (PCDA) monomers through spontaneous assembling onto the surface of superparamagnetic iron oxide (SPIO) nanoparticles to form micelles-polymerized structures. The results demonstrate that PDNCs will reduce the drug leakage and further control the drug release, and display self-responsive fluorescence upon intracellular uptake for cell trafficking and imaging-guided tumor treatment. The magnetic Lf-modified PDNCs with magnetic resonance imaging (MRI) and dual-targeting ability can enhance the transportation of the PDNCs across the BBB for tracking and targeting gliomas. An enhanced therapeutic efficiency can be obtained using Lf-Cur (Curcumin)-PDNCs by improving the retention time of the encapsulated Cur and producing fourfold higher Cur amounts in the brain compared to free Cur. Animal studies also confirm that Lf targeting and controlled release act synergistically to significantly suppress tumors in orthotopic brain-bearing rats. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Noninvasive quantitative measurement of colloid transport in mesoscale porous media using time lapse fluorescence imaging.

PubMed

Bridge, Jonathan W; Banwart, Steven A; Heathwaite, A Louise

2006-10-01

We demonstrate noninvasive quantitative imaging of colloid and solute transport at millimeter to decimeter (meso-) scale. Ultraviolet (UV) excited fluorescent solute and colloid tracers were independently measured simultaneously during co-advection through saturated quartz sand. Pulse-input experiments were conducted at constant flow rates and ionic strengths 10(-3), 10(-2) and 10(-1) M NaCl. Tracers were 1.9 microm carboxylate latex microspheres and disodium fluorescein. Spatial moments analysis was used to quantify relative changes in mass distribution of the colloid and solute tracers over time. The solute advected through the sand at a constant velocity proportional to flow rate and was described well by a conservative transport model (CXTFIT). In unfavorable deposition conditions increasing ionic strength produced significant reduction in colloid center of mass transport velocity over time. Velocity trends correlated with the increasing fraction of colloid mass retained along the flowpath. Attachment efficiencies (defined by colloid filtration theory) calculated from nondestructive retained mass data were 0.013 +/- 0.03, 0.09 +/- 0.02, and 0.22 +/- 0.05 at 10(-3), 10(-2), and 10(-1) M ionic strength, respectively, which compared well with previously published data from breakthrough curves and destructive sampling. Mesoscale imaging of colloid mass dynamics can quantify key deposition and transport parameters based on noninvasive, nondestructive, spatially high-resolution data.

Development of a General Aza-Cope Reaction Trigger Applied to Fluorescence Imaging of Formaldehyde in Living Cells.

PubMed

Bruemmer, Kevin J; Walvoord, Ryan R; Brewer, Thomas F; Burgos-Barragan, Guillermo; Wit, Niek; Pontel, Lucas B; Patel, Ketan J; Chang, Christopher J

2017-04-19

Formaldehyde (FA) is a reactive signaling molecule that is continuously produced through a number of central biological pathways spanning epigenetics to one-carbon metabolism. On the other hand, aberrant, elevated levels of FA are implicated in disease states ranging from asthma to neurodegenerative disorders. In this context, fluorescence-based probes for FA imaging are emerging as potentially powerful chemical tools to help disentangle the complexities of FA homeostasis and its physiological and pathological contributions. Currently available FA indicators require direct modification of the fluorophore backbone through complex synthetic considerations to enable FA detection, often limiting the generalization of designs to other fluorophore classes. To address this challenge, we now present the rational, iterative development of a general reaction-based trigger utilizing 2-aza-Cope reactivity for selective and sensitive detection of FA in living systems. Specifically, we developed a homoallylamine functionality that can undergo a subsequent self-immolative β-elimination, creating a FA-responsive trigger that is capable of masking a phenol on a fluorophore or any other potential chemical scaffold for related imaging and/or therapeutic applications. We demonstrate the utility of this trigger by creating a series of fluorescent probes for FA with excitation and emission wavelengths that span the UV to visible spectral regions through caging of a variety of dye units. In particular, Formaldehyde Probe 573 (FAP573), based on a resorufin scaffold, is the most red-shifted and FA sensitive in this series in terms of signal-to-noise responses and enables identification of alcohol dehydrogenase 5 (ADH5) as an enzyme that regulates FA metabolism in living cells. The results provide a starting point for the broader use of 2-aza-Cope reactivity for probing and manipulating FA biology.

Highly selective luminescent nanostructures for mitochondrial imaging and targeting

NASA Astrophysics Data System (ADS)

Fanizza, E.; Iacobazzi, R. M.; Laquintana, V.; Valente, G.; Caliandro, G.; Striccoli, M.; Agostiano, A.; Cutrignelli, A.; Lopedota, A.; Curri, M. L.; Franco, M.; Depalo, N.; Denora, N.

2016-02-01

Here a luminescent hybrid nanostructure based on functionalized quantum dots (QDs) is used as a fluorescent imaging agent able to target selectively mitochondria thanks to the molecular recognition of the translocator protein (TSPO). The selective targeting of such an 18 kDa protein mainly located in the outer mitochondrial membrane and overexpressed in several pathological states including neurodegenerative diseases and cancers may provide valuable information for the early diagnosis and therapy of human disorders. In particular, the rational design of amino functionalized luminescent silica coated QD nanoparticles (QD@SiO2 NPs) provides a versatile nanoplatform to anchor a potent and selective TSPO ligand, characterized by a 2-phenyl-imidazo[1,2-a]pyridine acetamide structure along with a derivatizable carboxylic end group, useful to conjugate the TSPO ligand and achieve TSPO-QD@SiO2 NPs by means of a covalent amide bond. The colloidal stability and optical properties of the proposed nanomaterials are comprehensively investigated and their potential as mitochondrial imaging agents is fully assessed. Sub-cellular fractionation, together with confocal laser scanning fluorescence microscopy and co-localization analysis of targeted TSPO-QD@SiO2 NPs in C6 glioma cells overexpressing the TSPO, proves the great potential of these multifunctional nanosystems as in vitro selective mitochondrial imaging agents.Here a luminescent hybrid nanostructure based on functionalized quantum dots (QDs) is used as a fluorescent imaging agent able to target selectively mitochondria thanks to the molecular recognition of the translocator protein (TSPO). The selective targeting of such an 18 kDa protein mainly located in the outer mitochondrial membrane and overexpressed in several pathological states including neurodegenerative diseases and cancers may provide valuable information for the early diagnosis and therapy of human disorders. In particular, the rational design of amino functionalized luminescent silica coated QD nanoparticles (QD@SiO2 NPs) provides a versatile nanoplatform to anchor a potent and selective TSPO ligand, characterized by a 2-phenyl-imidazo[1,2-a]pyridine acetamide structure along with a derivatizable carboxylic end group, useful to conjugate the TSPO ligand and achieve TSPO-QD@SiO2 NPs by means of a covalent amide bond. The colloidal stability and optical properties of the proposed nanomaterials are comprehensively investigated and their potential as mitochondrial imaging agents is fully assessed. Sub-cellular fractionation, together with confocal laser scanning fluorescence microscopy and co-localization analysis of targeted TSPO-QD@SiO2 NPs in C6 glioma cells overexpressing the TSPO, proves the great potential of these multifunctional nanosystems as in vitro selective mitochondrial imaging agents. Electronic supplementary information (ESI) available: Additional TEM micrographs, fluorescence and UV-Vis absorbance spectra of silica coated QD nanoparticles and TSPO ligand. See DOI: 10.1039/c5nr08139d

Fluorescence imaging spectroscopy (FIS) for comparing spectra from corn ears naturally and artificially infected with aflatoxin producing fungus.

PubMed

Hruska, Zuzana; Yao, Haibo; Kincaid, Russell; Darlington, Dawn; Brown, Robert L; Bhatnagar, Deepak; Cleveland, Thomas E

2013-08-01

In an effort to address the problem of rapid detection of aflatoxin in grain, particularly oilseeds, the current study assessed the spectral differences of aflatoxin production in kernels from a cornfield inoculated with spores from 2 different strains of toxigenic Aspergillus flavus. Aflatoxin production in corn from the same field due to natural infestation was also assessed. A small corn plot in Baton Rouge, La., U.S.A., was used during the 2008-growing season. Two groups of 400 plants were inoculated with 2 different inocula and 1 group of 400 plants was designated as controls. Any contamination detected in the controls was attributed to natural infestation. A subset of each group was imaged with a visible near infra red (VNIR) hyperspectral system under ultra violet (UV) excitation and subsequently analyzed for aflatoxin using affinity column fluorometry. Group differences were statistically analyzed. Results indicate that when all the spectral data across all groups were averaged, any potential differences between groups (treated and untreated) were obscured. However, spectral analysis based on contaminated "hot" pixel classification showed a distinct spectral shift/separation between contaminated and clean ears with fluorescence peaks at 501 and 478 nm, respectively. All inoculated and naturally infected control ears had fluorescence peaks at 501 nm that differed from uninfected corn ears. Results from this study may be useful in evaluating rapid, noninvasive instrumentation and/or methodology for aflatoxin detection in grain. © 2013 Institute of Food Technologists®

A portable near-infrared fluorescence image overlay device for surgical navigation (Conference Presentation)

NASA Astrophysics Data System (ADS)

McWade, Melanie A.

2016-03-01

A rise in the use of near-infrared (NIR) fluorescent dyes or intrinsic fluorescent markers for surgical guidance and tissue diagnosis has triggered the development of NIR fluorescence imaging systems. Because NIR wavelengths are invisible to the naked eye, instrumentation must allow surgeons to visualize areas of high fluorescence. Current NIR fluorescence imaging systems have limited ease-of-use because they display fluorescent information on remote display monitors that require surgeons to divert attention away from the patient to identify the location of tissue fluorescence. Furthermore, some systems lack simultaneous visible light imaging which provides valuable spatial context to fluorescence images. We have developed a novel, portable NIR fluorescence imaging approach for intraoperative surgical guidance that provides information for surgical navigation within the clinician's line of sight. The system utilizes a NIR CMOS detector to collect excited NIR fluorescence from the surgical field. Tissues with NIR fluorescence are overlaid with visible light to provide information on tissue margins directly on the surgical field. In vitro studies have shown this versatile imaging system can be applied to applications with both extrinsic NIR contrast agents such as indocyanine green and weaker sources of biological fluorescence such as parathyroid gland tissue. This non-invasive, portable NIR fluorescence imaging system overlays an image directly on tissue, potentially allowing surgical decisions to be made quicker and with greater ease-of-use than current NIR fluorescence imaging systems.

Impact of metal binding on the antitumor activity and cellular imaging of a metal chelator cationic imidazopyridine derivative.

PubMed

Roy, Mithun; Chakravarthi, Balabhadrapatruni V S K; Jayabaskaran, Chelliah; Karande, Anjali A; Chakravarty, Akhil R

2011-05-14

A new water soluble cationic imidazopyridine species, viz. (1E)-1-((pyridin-2-yl)methyleneamino)-3-(3-(pyridin-2-yl)imidazo[1,5-a]pyridin-2(3H)-yl)propan-2-ol (1), as a metal chelator is prepared as its PF(6) salt and characterized. Compound 1 shows fluorescence at 438 nm on excitation at 342 nm in Tris-HCl buffer giving a fluorescence quantum yield (φ) of 0.105 and a life-time of 5.4 ns. Compound 1, as an avid DNA minor groove binder, shows pUC19 DNA cleavage activity in UV-A light of 365 nm forming singlet oxygen species in a type-II pathway. The photonuclease potential of 1 gets enhanced in the presence of Fe(2+), Cu(2+) or Zn(2+). Compound 1 itself displays anticancer activity in HeLa, HepG2 and Jurkat cells with an enhancement on addition of the metal ions. Photodynamic effect of 1 at 365 nm also gets enhanced in the presence of Fe(2+) and Zn(2+). Fluorescence-based cell cycle analysis shows a significant dead cell population in the sub-G1 phase of the cell cycle suggesting apoptosis via ROS generation. A significant change in the nuclear morphology is observed from Hoechst 33258 and an acridine orange/ethidium bromide (AO/EB) dual nuclear staining suggesting apoptosis in cells when treated with 1 alone or in the presence of the metal ions. Apoptosis is found to be caspase-dependent. Fluorescence imaging to monitor the distribution of 1 in cells shows that 1 in the presence of metal ions accumulates predominantly in the cytoplasm. Enhanced uptake of 1 into the cells within 12 h is observed in the presence of Fe(2+) and Zn(2+).

qF-SSOP: real-time optical property corrected fluorescence imaging

PubMed Central

Valdes, Pablo A.; Angelo, Joseph P.; Choi, Hak Soo; Gioux, Sylvain

2017-01-01

Fluorescence imaging is well suited to provide image guidance during resections in oncologic and vascular surgery. However, the distorting effects of tissue optical properties on the emitted fluorescence are poorly compensated for on even the most advanced fluorescence image guidance systems, leading to subjective and inaccurate estimates of tissue fluorophore concentrations. Here we present a novel fluorescence imaging technique that performs real-time (i.e., video rate) optical property corrected fluorescence imaging. We perform full field of view simultaneous imaging of tissue optical properties using Single Snapshot of Optical Properties (SSOP) and fluorescence detection. The estimated optical properties are used to correct the emitted fluorescence with a quantitative fluorescence model to provide quantitative fluorescence-Single Snapshot of Optical Properties (qF-SSOP) images with less than 5% error. The technique is rigorous, fast, and quantitative, enabling ease of integration into the surgical workflow with the potential to improve molecular guidance intraoperatively. PMID:28856038

Quantum dots versus organic fluorophores in fluorescent deep-tissue imaging--merits and demerits.

PubMed

Bakalova, Rumiana; Zhelev, Zhivko; Gadjeva, Veselina

2008-12-01

The use of fluorescence in deep-tissue imaging is rapidly expanding in last several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecular targets in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With the development of novel bright fluorophores based on nanotechnologies and 3D fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. The fluorescent imaging has also a potential to give a real map of human anatomy and physiology. The current review outlines the advantages of fluorescent nanoparticles over conventional organic dyes in deep-tissue imaging in vivo and defines the major requirements to the "perfect fluorophore". The analysis proceeds from the basic principles of fluorescence and major characteristics of fluorophores, light-tissue interactions, and major limitations of fluorescent deep-tissue imaging. The article is addressed to a broad readership - from specialists in this field to university students.

Enhancement of growth and lipid production from microalgae using fluorescent paint under the solar radiation.

PubMed

Seo, Yeong Hwan; Cho, Changsoon; Lee, Jung-Yong; Han, Jong-In

2014-12-01

Solar radiation has intensity that is too high to inhibit microalgae activity and is composed of wide light spectrum including ultraviolet (UV) range which cannot be utilized for microalgae. For these reasons, the modification of solar radiation is required for effective microalgae cultivation, and to do that, fluorescent paint was used for not only blocking excessive solar energy but also converting UV to visible light. With fluorescent aqueous layer, microalgae was protected from photoinhibition and could grow well, but there was difference in growth and lipid accumulation efficiencies depending on the color; maximum dry weight of 1.7 g/L was achieved in red paint, whereas best lipid content of 30% was obtained in blue one. This phenomenon was due to the different light spectrum made by colors. With simple process using fluorescent paint, modification of light was successfully done and allowing microalgae to grow under strong radiation such as solar radiation. Copyright © 2014 Elsevier Ltd. All rights reserved.

[The spectroscopic studies on the binding of Al(III) to EHPG].

PubMed

Li, Ying-qi; Bai, Hai-jing; Yang, Bin-sheng

2002-06-01

Interaction of ethylene-N,N'-bis(o-hydioxyphenylglycine) (EHPG) with Al3+ has been investigated by both UV difference and fluorescent spectra. Both results show that the molar ration of the complex is most likely 1:1. Aluminum binding produces peaks at 235 and 291 nm. The molar absorptivity of aluminum ions to EHPG at 235 nm is 1.27 x 10(4) cm-1.mol-1.L. The conditional stability constant for Al3+ binding to EHPG is determined to be IgK = 14.20 +/- 0.03 in 0.1 mol.L-1 Hepes buffer at room temperature, pH 7.4 by UV difference spectra. At the same condition, the fluorescent intensity of EHPG at 310 nm has been monitored. In result, the fluorescent intensity of EHPG at 310 nm is decreased with the addition of Al3+. Then the quench of the fluorescent intensity is ascribed to deprotonated phenolic groups coordinated to aluminum ions.

Spectrometric studies on the interaction of fluoroquinolones and bovine serum albumin

NASA Astrophysics Data System (ADS)

Ni, Yongnian; Su, Shaojing; Kokot, Serge

2010-02-01

The interaction between fluoroquinolones (FQs), ofloxacin and enrofloxacin, and bovine serum albumin (BSA) was investigated by fluorescence and UV-vis spectroscopy. It was demonstrated that the fluorescence quenching of BSA by FQ is a result of the formation of the FQ-BSA complex stabilized, in the main, by hydrogen bonds and van der Waals forces. The Stern-Volmer quenching constant, KSV, and the corresponding thermodynamic parameters, Δ H, Δ S and Δ G, were estimated. The distance, r, between the donor, BSA, and the acceptor, FQ, was estimated from fluorescence resonance energy transfer (FRET). The effect of FQ on the conformation of BSA was analyzed with the aid of UV-vis absorbance spectra and synchronous fluorescence spectroscopy. Spectral analysis showed that the two FQs affected the conformation of the BSA but in a different manner. Thus, with ofloxacin, the polarity around the tryptophan residues decreased and the hydrophobicity increased, while for enrofloxacin, the opposite effect was observed.

Serious complications in experiments in which UV doses are effected by using different lamp heights.

PubMed

Flint, Stephan D; Ryel, Ronald J; Hudelson, Timothy J; Caldwell, Martyn M

2009-10-06

Many experiments examining plant responses to enhanced ultraviolet-B radiation (280-315nm) simply compare an enhanced UV-B treatment with ambient UV-B (or no UV-B radiation in most greenhouse and controlled-environment studies). Some more detailed experiments utilize multiple levels of UV-B radiation. A number of different techniques have been used to adjust the UV dose. One common technique is to place racks of fluorescent UV-emitting lamps at different heights above the plant canopy. However, the lamps and associated support structure cast shadows on the plant bed below. We calculated one example of the sequence of shade intervals for two common heights of lamp racks and show the patterns and duration of shade which the plants receive is distributed differently over the course of the day for different heights of the lamp racks. We also conducted a greenhouse experiment with plants (canola, sunflower and maize) grown under unenergized lamp racks suspended at the same two heights above the canopy. Growth characteristics differed in unpredictable ways between plants grown under the two heights of lamp racks. These differences could enhance or obscure potential UV-B effects. Also, differences in leaf mass per unit foliage area, which were observed in this experiment, could contribute to differences in plant UV-B sensitivity. We recommend the use of other techniques for achieving multiple doses of UV-B radiation. These range from simple and inexpensive approaches (e.g., wrapping individual fluorescent tubes in layers of a neutral-density filter such as cheese cloth) to more technical and expensive alternatives (e.g., electronically modulated lamp control systems). These choices should be determined according to the goals of the particular experiment.

Exclusion of solar UV radiation improves photosynthetic performance and yield of wheat varieties.

PubMed

Kataria, Sunita; Guruprasad, K N

2015-12-01

Field studies were conducted to determine the potential for alterations in photosynthetic performance and grain yield of four wheat (Triticum aestivum) varieties of India- Vidisha, Purna, Swarna and Naveen Chandausi by ambient ultraviolet radiation (UV). The plants were grown in specially designed UV exclusion chambers, wrapped with filters that excluded UV-B (<315 nm), UV-A/B (<400 nm) or transmitted ambient UV or lacked filters. The results indicated that solar UV exclusion increased the leaf mass per area ratio, leaf weight ratio and chlorophylls per unit area of flag leaves in all the four varieties of wheat. Polyphasic chlorophyll a fluorescence transients from the flag leaves of UV excluded wheat plants gave a higher fluorescence yield. Exclusion of solar UV significantly enhanced photosynthetic performance as a consequence of increased efficiency of PS II, performance index (PIABS) and rate of photosynthesis in the flag leaves of wheat varieties along with a remarkable increase in carbonic anhydrase, Rubisco and nitrate reductase activities. This additional fixation of carbon and nitrogen by exclusion of UV was channelized towards the improvement in grain yield of wheat varieties as there was a decrease in the UV-B absorbing substances and an increase in soluble protein content in flag leaves of all the four varieties of wheat. The magnitude of response for UV exclusion for all the measured parameters was higher in two varieties of wheat Vidisha and Purna as compared to Swarna and Naveen Chandausi. Cumulative stress response index (CSRI) for each variety was developed from the cumulative sum of physiological and yield parameters such as leaf mass area ratio of flag leaf, total chlorophyll content, performance index at absorption basis, rate of photosynthesis and grain yield. All the varieties had a negative CSRI, demonstrating a negative impact of ambient UV radiation. Naveen Chandausi and Swarna are less sensitive to ambient UV radiation; Vidisha is more sensitive to both UV-A and UV-B and Purna is more sensitive to ambient UV-B radiation. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Extending the use of ultraviolet light for fruit quality sorting in citrus packinghouses

USDA-ARS?s Scientific Manuscript database

Illumination with ultraviolet light (UV) is commonly used in citrus packinghouses as a means to aid in the identification and removal of decayed oranges from the packline. This technique is effective because areas of decay strongly fluoresce under UV illumination. It was observed that oranges often ...

In Vivo Follow-up of Brain Tumor Growth via Bioluminescence Imaging and Fluorescence Tomography

PubMed Central

Genevois, Coralie; Loiseau, Hugues; Couillaud, Franck

2016-01-01

Reporter gene-based strategies are widely used in experimental oncology. Bioluminescence imaging (BLI) using the firefly luciferase (Fluc) as a reporter gene and d-luciferin as a substrate is currently the most widely employed technique. The present paper compares the performances of BLI imaging with fluorescence imaging using the near infrared fluorescent protein (iRFP) to monitor brain tumor growth in mice. Fluorescence imaging includes fluorescence reflectance imaging (FRI), fluorescence diffuse optical tomography (fDOT), and fluorescence molecular Imaging (FMT®). A U87 cell line was genetically modified for constitutive expression of both the encoding Fluc and iRFP reporter genes and assayed for cell, subcutaneous tumor and brain tumor imaging. On cultured cells, BLI was more sensitive than FRI; in vivo, tumors were first detected by BLI. Fluorescence of iRFP provided convenient tools such as flux cytometry, direct detection of the fluorescent protein on histological slices, and fluorescent tomography that allowed for 3D localization and absolute quantification of the fluorescent signal in brain tumors. PMID:27809256

In Vivo Follow-up of Brain Tumor Growth via Bioluminescence Imaging and Fluorescence Tomography.

PubMed

Genevois, Coralie; Loiseau, Hugues; Couillaud, Franck

2016-10-31

Reporter gene-based strategies are widely used in experimental oncology. Bioluminescence imaging (BLI) using the firefly luciferase (Fluc) as a reporter gene and d-luciferin as a substrate is currently the most widely employed technique. The present paper compares the performances of BLI imaging with fluorescence imaging using the near infrared fluorescent protein (iRFP) to monitor brain tumor growth in mice. Fluorescence imaging includes fluorescence reflectance imaging (FRI), fluorescence diffuse optical tomography (fDOT), and fluorescence molecular Imaging (FMT ® ). A U87 cell line was genetically modified for constitutive expression of both the encoding Fluc and iRFP reporter genes and assayed for cell, subcutaneous tumor and brain tumor imaging. On cultured cells, BLI was more sensitive than FRI; in vivo, tumors were first detected by BLI. Fluorescence of iRFP provided convenient tools such as flux cytometry, direct detection of the fluorescent protein on histological slices, and fluorescent tomography that allowed for 3D localization and absolute quantification of the fluorescent signal in brain tumors.

White-nose syndrome pathology grading in Nearctic and Palearctic bats

PubMed Central

Pikula, Jiri; Amelon, Sybill K.; Bandouchova, Hana; Bartonička, Tomáš; Berkova, Hana; Brichta, Jiri; Hooper, Sarah; Kokurewicz, Tomasz; Kolarik, Miroslav; Köllner, Bernd; Kovacova, Veronika; Linhart, Petr; Piacek, Vladimir; Turner, Gregory G.; Zukal, Jan; Martínková, Natália

2017-01-01

While white-nose syndrome (WNS) has decimated hibernating bat populations in the Nearctic, species from the Palearctic appear to cope better with the fungal skin infection causing WNS. This has encouraged multiple hypotheses on the mechanisms leading to differential survival of species exposed to the same pathogen. To facilitate intercontinental comparisons, we proposed a novel pathogenesis-based grading scheme consistent with WNS diagnosis histopathology criteria. UV light-guided collection was used to obtain single biopsies from Nearctic and Palearctic bat wing membranes non-lethally. The proposed scheme scores eleven grades associated with WNS on histopathology. Given weights reflective of grade severity, the sum of findings from an individual results in weighted cumulative WNS pathology score. The probability of finding fungal skin colonisation and single, multiple or confluent cupping erosions increased with increase in Pseudogymnoascus destructans load. Increasing fungal load mimicked progression of skin infection from epidermal surface colonisation to deep dermal invasion. Similarly, the number of UV-fluorescent lesions increased with increasing weighted cumulative WNS pathology score, demonstrating congruence between WNS-associated tissue damage and extent of UV fluorescence. In a case report, we demonstrated that UV-fluorescence disappears within two weeks of euthermy. Change in fluorescence was coupled with a reduction in weighted cumulative WNS pathology score, whereby both methods lost diagnostic utility. While weighted cumulative WNS pathology scores were greater in the Nearctic than Palearctic, values for Nearctic bats were within the range of those for Palearctic species. Accumulation of wing damage probably influences mortality in affected bats, as demonstrated by a fatal case of Myotis daubentonii with natural WNS infection and healing in Myotis myotis. The proposed semi-quantitative pathology score provided good agreement between experienced raters, showing it to be a powerful and widely applicable tool for defining WNS severity. PMID:28767673

White-nose syndrome pathology grading in Nearctic and Palearctic bats.

PubMed

Pikula, Jiri; Amelon, Sybill K; Bandouchova, Hana; Bartonička, Tomáš; Berkova, Hana; Brichta, Jiri; Hooper, Sarah; Kokurewicz, Tomasz; Kolarik, Miroslav; Köllner, Bernd; Kovacova, Veronika; Linhart, Petr; Piacek, Vladimir; Turner, Gregory G; Zukal, Jan; Martínková, Natália

2017-01-01

While white-nose syndrome (WNS) has decimated hibernating bat populations in the Nearctic, species from the Palearctic appear to cope better with the fungal skin infection causing WNS. This has encouraged multiple hypotheses on the mechanisms leading to differential survival of species exposed to the same pathogen. To facilitate intercontinental comparisons, we proposed a novel pathogenesis-based grading scheme consistent with WNS diagnosis histopathology criteria. UV light-guided collection was used to obtain single biopsies from Nearctic and Palearctic bat wing membranes non-lethally. The proposed scheme scores eleven grades associated with WNS on histopathology. Given weights reflective of grade severity, the sum of findings from an individual results in weighted cumulative WNS pathology score. The probability of finding fungal skin colonisation and single, multiple or confluent cupping erosions increased with increase in Pseudogymnoascus destructans load. Increasing fungal load mimicked progression of skin infection from epidermal surface colonisation to deep dermal invasion. Similarly, the number of UV-fluorescent lesions increased with increasing weighted cumulative WNS pathology score, demonstrating congruence between WNS-associated tissue damage and extent of UV fluorescence. In a case report, we demonstrated that UV-fluorescence disappears within two weeks of euthermy. Change in fluorescence was coupled with a reduction in weighted cumulative WNS pathology score, whereby both methods lost diagnostic utility. While weighted cumulative WNS pathology scores were greater in the Nearctic than Palearctic, values for Nearctic bats were within the range of those for Palearctic species. Accumulation of wing damage probably influences mortality in affected bats, as demonstrated by a fatal case of Myotis daubentonii with natural WNS infection and healing in Myotis myotis. The proposed semi-quantitative pathology score provided good agreement between experienced raters, showing it to be a powerful and widely applicable tool for defining WNS severity.

Fluorescence Imaging of Underexpanded Jets and Comparison with CFD

NASA Technical Reports Server (NTRS)

Wilkes, Jennifer A.; Glass, Christopher E.; Danehy, Paul M.; Nowak, Robert J.

2006-01-01

An experimental study of underexpanded and highly underexpanded axisymmetric nitrogen free jets seeded with 0.5% nitric oxide (NO) and issuing from a sonic orifice was conducted at NASA Langley Research Center. Reynolds numbers based on nozzle exit conditions ranged from 770 to 35,700, and nozzle exit-to-ambient jet pressure ratios ranged from 2 to 35. These flows were non-intrusively visualized with a spatial resolution of approximately 0.14 mm x 0.14 mm x 1 mm thick and a temporal resolution of 1 s using planar laser-induced fluorescence (PLIF) of NO, with the laser tuned to the strongly-fluorescing UV absorption bands of the Q1 band head near 226.256 nm. Three laminar cases were selected for comparison with computational fluid dynamics (CFD). The cases were run using GASP (General Aerodynamic Simulation Program) Version 4. Comparisons of the fundamental wavelength of the jet flow showed good agreement between CFD and experiment for all three test cases, while comparisons of Mach disk location and Mach disk diameter showed good agreement at lower jet pressure ratios, with a tendency to slightly underpredict these parameters with increasing jet pressure ratio.

Fluorescein diacetate for determination of cell viability in 3D fibroblast-collagen-GAG constructs.

PubMed

Powell, Heather M; Armour, Alexis D; Boyce, Steven T

2011-01-01

Quantification of cell viability and distribution within engineered tissues currently relies on representative histology, phenotypic assays, and destructive assays of viability. To evaluate uniformity of cell density throughout 3D collagen scaffolds prior to in vivo use, a nondestructive, field assessment of cell viability is advantageous. Here, we describe a field measure of cell viability in lyophilized collagen-glycosaminoglycan (C-GAG) scaffolds in vitro using fluorescein diacetate (FdA). Fibroblast-C-GAG constructs are stained 1 day after cellular inoculation using 0.04 mg/ml FdA followed by exposure to 366 nm UV light. Construct fluorescence quantified using Metamorph image analysis is correlated with inoculation density, MTT values, and histology of corresponding biopsies. Construct fluorescence correlates significantly with inoculation density (p  <  0.001) and MTT values (p  <  0.001) of biopsies collected immediately after FdA staining. No toxicity is detected in the constructs, as measured by MTT assay before and after the FdA assay at different time points; normal in vitro histology is demonstrated for the FdA-exposed constructs. In conclusion, measurement of intracellular fluorescence with FdA allows for the early, comprehensive measurement of cellular distributions and viability in engineered tissue.

Measurement of liquid film thickness by optical fluorescence and its application to an oscillating piston positive displacement flowmeter

NASA Astrophysics Data System (ADS)

Morton, Charlotte E.; Baker, Roger C.; Hutchings, Ian M.

2011-12-01

The movement of the circular piston in an oscillating piston positive displacement flowmeter is important in understanding the operation of the flowmeter, and the leakage of liquid past the piston plays a key role in the performance of the meter. The clearances between the piston and the chamber are small, typically less than 60 µm. In order to measure this film thickness a fluorescent dye was added to the water passing through the meter, which was illuminated with UV light. Visible light images were captured with a digital camera and analysed to give a measure of the film thickness with an uncertainty of less than 7%. It is known that this method lacks precision unless careful calibration is undertaken. Methods to achieve this are discussed in the paper. The grey level values for a range of film thicknesses were calibrated in situ with six dye concentrations to select the most appropriate one for the range of liquid film thickness. Data obtained for the oscillating piston flowmeter demonstrate the value of the fluorescence technique. The method is useful, inexpensive and straightforward and can be extended to other applications where measurement of liquid film thickness is required.

Fabrication, Light Emission, and Magnetism of Silica Nanoparticles Hybridized with AIE Luminogens and Inorganic Nanostructures

NASA Astrophysics Data System (ADS)

Faisal, Mahtab

Much research efforts have been devoted in developing new synthetic approaches for fluorescent silica nanoparticles (FSNPs) due to their potential high-technological applications. However, light emissions from most of the FSNPs prepared so far have been rather weak. This is due to the emission quenching caused by the aggregation of fluorophores in the solid state. We have observed a novel phenomenon of aggregation-induced emission (AIE): a series of propeller-shaped molecules such as tetraphenylethene (TPE) and silole are induced to emit efficiently by aggregate formation. Thus, they are ideal fluorophors for the construction of FSNPs and my thesis work focuses on the synthesis of silica nanoparticles containing these luminogens and magnetic nanostructures. Highly emissive FSNPs with core-shell structures are fabricated by surfactant-free sol-gel reactions of tetraphenylethene- (TPE) and silole-functionalized siloxanes followed by the reactions with tetraethoxysilane. The FSNPs are uniformly sized, surface-charged and colloidally stable. The diameters of the FSNPs are tunable in the range of 45--295 nm by changing the reaction conditions. Whereas their TPE and silole precursors are non-emissive, the FSNPs emit strong visible lights, thanks to the novel aggregation-induced emission characteristics of the TPE and silole aggregates in the hybrid nanoparticles. The FSNPs pose no toxicity to living cells and can be utilized to selectively image cytoplasm of HeLa cells. Applying the same tool in the presence of citrate-coated magnetite nanoparticles, uniform magnetic fluorescent silica nanoparticles (MFSNPs) with smooth surfaces are fabricated. These particles exhibit appreciable surface charges and hence good colloidal stability. They are superparamagnetic, exhibiting no hysteresis at room temperature. UV irradiation of a suspension of MFSNPs in ethanol gives strong blue and green emissions. The MFSNPs can selectively stain the cytoplasmic regions of the living cells. Sol-gel reaction in the presence of (3-aminopropyl)triethoxysilane has generated MFSNP-NH2 with numerous amino functionalities decorated on the surfaces, enabling them to immobilize bovine serum albumin efficiently. FSNPs with strong light emissions are facilely fabricated by thio-click chemistry, Cu(I)-catalyzed 1,3-dipolar cycloaddition, and sol-gel reaction. The FSNPs are characterized by SEM, TEM, IR, PL, and zeta potential analyses. They are uniformly sized with smooth surfaces. Upon photoexcitation, the FSNPs emit strong visible lights with fluorescence quantum yields up to 25.5%. Sugar-functionalized fluorescent silica nanoparticles are facilely fabricated by click reaction of azide-modified FSNPs with sugar- containing phenylacetylene catalyzed by Cu(PPh3)3Br in THF. The nanoparticles are uniformly sized and emit efficient light upon photoexcitation. They can function as fluorescent visualizers for intracellular imaging and can target specific cancer cells. Folic acid-functionalized fluorescent silica nanoparticles are facilely fabricated by surface functionalization of FSNPs with folic acid. The nanoparticles are spherical in shape. They possess high zeta potentials and hence exhibit excellent colloidal stability. UV irradiation of suspensions of the nanoparticles in ethanol gives strong blue and green emissions at 465 and 490 nm with absolute fluorescence quantum yields up to 47%. Carboxylic acid and thiol-functionalized fluorescent silica nanoparticles (FSNP-COOH and FSNP-SH) with uniform particle sizes, narrow size distributions, and smooth surface morphologies are fabricated. The nanoparticles possess high surface charges and exhibit strong light emissions upon photoexcitation. They can adsorb lysozyme strongly on their surfaces and for 5 mg of FSNP-COOH and FSNP-SH, they can take 209 and 86 mug of lysozyme. Thus, they are potential carriers for protein and fluorescent probes or biosensors for an array of biological applications.

Carcinogenic damage to deoxyribonucleic acid is induced by near-infrared laser pulses in multiphoton microscopy via combination of two- and three-photon absorption

NASA Astrophysics Data System (ADS)

Nadiarnykh, Oleg; Thomas, Giju; Van Voskuilen, Johan; Sterenborg, Henricus J. C. M.; Gerritsen, Hans C.

2012-11-01

Nonlinear optical imaging modalities (multiphoton excited fluorescence, second and third harmonic generation) applied in vivo are increasingly promising for clinical diagnostics and the monitoring of cancer and other disorders, as they can probe tissue with high diffraction-limited resolution at near-infrared (IR) wavelengths. However, high peak intensity of femtosecond laser pulses required for two-photon processes causes formation of cyclobutane-pyrimidine-dimers (CPDs) in cellular deoxyribonucleic acid (DNA) similar to damage from exposure to solar ultraviolet (UV) light. Inaccurate repair of subsequent mutations increases the risk of carcinogenesis. In this study, we investigate CPD damage that results in Chinese hamster ovary cells in vitro from imaging them with two-photon excited autofluorescence. The CPD levels are quantified by immunofluorescent staining. We further evaluate the extent of CPD damage with respect to varied wavelength, pulse width at focal plane, and pixel dwell time as compared with more pronounced damage from UV sources. While CPD damage has been expected to result from three-photon absorption, our results reveal that CPDs are induced by competing two- and three-photon absorption processes, where the former accesses UVA absorption band. This finding is independently confirmed by nonlinear dependencies of damage on laser power, wavelength, and pulse width.

One trinucleus dimethine cyanine dye: Experimental and theoretical studies on molecular structure as well as absorption and fluorescence properties

NASA Astrophysics Data System (ADS)

Zhang, D. D.; Wang, L. Y.; Su, J. J.; Zhang, X. F.; Lei, Y. B.; Zhai, G. H.; Wen, Z. Y.

2013-05-01

A kind of trinucleus dimethine cyanine dye: 1-methyl-2,6-bis[2-(furan-2-yl)vinyl]pyridinium iodide (1) was synthesized and characterized by 1H NMR, 13C NMR, IR, MS, UV-Vis spectroscopy and elemental analysis. The crystals of dye 1, obtained from slow evaporation of solvent acetone, crystallized in the triclinic space group P - 1 with a = 9.6501(16) Å, b = 10.2308(17) Å, c = 10.7341(17) Å, V = 887.2(3) Å3, and Z = 2 (at 298(2) K), and it was stabilized by the hydrogen bonds and intermolecular face-to-face π⋯π aromatic stacking interactions. Crystallographic, IR, 1H NMR and UV-Vis data of dye 1 were compared with the results of density functional theory (DFT) method, and the calculated molecular geometries, vibrational bands, 1H NMR chemical shifts and UV-Vis maximum absorption were consistent with the experimental results. The fluorescence spectra were predicted in four different solvents with CIS/PCM methods. Compared with experimental values, the absolute deviations of emission maxima were -17.4 nm in chloroform, 6.3 nm in DMSO, 4.9 nm in methanol, and 6.8 nm in water, respectively. And the experimental fluorescence spectra were nicely reproduced by the simulated fluorescence spectra for each solvent.

UV and fluorescence spectral changes induced by neodymium binding of N,N'-ethylenebis[2-(o-hydroxyphenolic)glycine] and N,N'-di(2-hydroxybenzyl)ethylenediamine-N,N' diacetic acid.

PubMed

Wang, Zhijun; Yang, Binsheng

2006-11-01

In 0.01 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Hepes), pH 7.4 and room temperature, the binding of neodymium to N,N'-ethylenebis[2-(o-hydroxyphenolic)glycine] (EHPG), or N,N'-di(2-hydroxybenzyl)ethylenediamine-N,N' diacetic acid (HBED) had been studied from 210 to 330 nm by means of difference UV spectra. Two peaks at 240 and 292 nm appear in difference UV spectra after neodymium binding to EHPG or HBED. The 1:1 stable complex can be confirmed from spectral titration curves. The molar extinction coefficient of Nd-EHPG and Nd-HBED complexes are Deltaepsilon(Nd-EHPG)=(12.93+/-0.21) x 10(3)cm(-1)M(-1), Deltaepsilon(Nd-HBED)=(14.45+/-0.51) x 10(5)cm(-1)M(-1) at 240 nm, respectively. Using EDTA as a competitor, the conditional equilibrium constants of the complexes are logK(Nd-EHPG)=11.89+/-0.09 and logK(Nd-HBED)=12.19+/-0.15, respectively. At the same conditions, fluorescence measurements show that neodymium binding to EHPG leads to a quenching of the fluorescence of EHPG at near 310 nm. However, there is no obvious fluorescence change of HBED at 318 nm with the binding of neodymium to HBED.

Detection of biological warfare agents using ultra violet-laser induced fluorescence LIDAR

NASA Astrophysics Data System (ADS)

Joshi, Deepti; Kumar, Deepak; Maini, Anil K.; Sharma, Ramesh C.

This review has been written to highlight the threat of biological warfare agents, their types and detection. Bacterial biological agent Bacillus anthracis (bacteria causing the disease anthrax) which is most likely to be employed in biological warfare is being discussed in detail. Standoff detection of biological warfare agents in aerosol form using Ultra violet-Laser Induced Fluorescence (UV-LIF) spectroscopy method has been studied. Range-resolved detection and identification of biological aerosols by both nano-second and non-linear femto-second LIDAR is also discussed. Calculated received fluorescence signal for a cloud of typical biological agent Bacillus globigii (Simulants of B. anthracis) at a location of ˜5.0 km at different concentrations in presence of solar background radiation has been described. Overview of current research efforts in internationally available working UV-LIF LIDAR systems are also mentioned briefly.

Near-UV OH Prompt Emission in the Innermost Coma of 103P/Hartley 2

NASA Astrophysics Data System (ADS)

La Forgia, Fiorangela; Bodewits, Dennis; A'Hearn, Michael F.; Protopapa, Silvia; Kelley, Michael S. P.; Sunshine, Jessica; Feaga, Lori; Farnham, Tony

2017-11-01

The Deep Impact spacecraft flyby of comet 103P/Hartley 2 occurred on 2010 November 4, 1 week after perihelion with a closest approach (CA) distance of about 700 km. We used narrowband images obtained by the Medium Resolution Imager on board the spacecraft to study the gas and dust in the innermost coma. We derived an overall dust reddening of 15%/100 nm between 345 and 749 nm and identified a blue enhancement in the dust coma in the sunward direction within 5 km from the nucleus, which we interpret as a localized enrichment in water ice. OH column density maps show an anti-sunward enhancement throughout the encounter, except for the highest-resolution images, acquired at CA, where a radial jet becomes visible in the innermost coma, extending up to 12 km from the nucleus. The OH distribution in the inner coma is very different from that expected for a fragment species. Instead, it correlates well with the water vapor map derived by the HRI-IR instrument on board Deep Impact. Radial profiles of the OH column density and derived water production rates show an excess of OH emission during CA that cannot be explained with pure fluorescence. We attribute this excess to a prompt emission process where photodissociation of H2O directly produces excited OH*(A 2Σ+) radicals. Our observations provide the first direct imaging of near-UV prompt emission of OH. We therefore suggest the use of a dedicated filter centered at 318.8 nm to directly trace the water in the coma of comets.

In Situ Determination of Illegal Drugs in Oral Fluid by Portable Capillary Electrophoresis with Deep UV Excited Fluorescence Detection.

PubMed

Saar-Reismaa, Piret; Erme, Enn; Vaher, Merike; Kulp, Maria; Kaljurand, Mihkel; Mazina-Šinkar, Jekaterina

2018-05-15

The present study demonstrates the potential of a portable capillary electrophoresis (CE) instrument, coupled to deep UV fluorescence detector (FD) with a 230-255 nm excitation wavelength range, for the determination of the abuse of illegal drugs in oral fluids in situ. CE was introduced in this study due its exceptional power of separation and resolution, short analysis time, and ability for miniaturization for on-site assessment of different substances. The deep UV fluorescence detector was equipped with five interchangeable emission filters, in the emission wavelength range from 278 to 600 nm, and was successfully employed for determination of natively fluorescing illegal drugs, such as cocaine, cocaethylene, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxeamphetamine (MDA), 3,4-methylenedioxy- N-ethylamphetamine (MDEA), para-methoxyamphetamine (PMA), para-methoxy- N-methylamphetamine (PMMA), amphetamine (AMP), methamphetamine (METH), tetrahydrocannabinol (THC) and cannabidiol (CBD). The developed FD showed impressive sensitivity. The instrumental detection limit was 0.5 μg/L for MDMA. It also showed broad linearity, up to 50 mg/L for MDMA. The noise CV% was 1.1% for an empty capillary and 0.6% for a capillary filled with acetonitrile. The portable CE-FD with developed electrophoretic methodologies was successfully utilized for the determination of illegal abuse of drugs during "Weekend" 2016 and 2017 Music Festivals (Estonia). Moreover, CE-FD can be employed for detection of other natively fluorescing compounds in the proposed range (e.g., for different phenolic compounds, BTEX, naphthalene derivatives, and others), significantly widening the applicability of developed CE-FD instrument.

Development of green fluorescent protein-expressing bacterial strains and evaluation for potential use as positive controls in sample analyses.

PubMed

Noah, Charles W; Shaw, Christine I; Ikeda, Jack S; Kreuzer, Karen S; Sofos, John N

2005-04-01

Strains of enterohemorrhagic Escherichia coli O157:H7 and Salmonella Typhimurium were engineered to express the gene for a modified green fluorescent protein (GFP) and were evaluated for potential use as positive controls in sample analyses. The strains fluoresced when observed as colonies with a handheld UV lamp or as individual cells under a fluorescent microscope. The strains maintained their fluorescence following growth in three series of transfer experiments including 8 to 11 passages from broth to broth and twice for 15 consecutive transfers from broth onto Trypticase soy agar plates. Cultures also maintained stability in the ability to fluoresce when agar plates were refrigerated (4 degrees C) for up to 12 days. Growth characteristics of the GFP-positive strains were comparable to those of corresponding control strains. The GFP-positive strains were successfully identified using rapid diagnostic methods and were differentiated from their corresponding non-GFP strains by pulsed-field gel electrophoresis but not by repetitive extragenic palindromic PCR. The GFP-positive and the control strains were recovered successfully from individually inoculated food samples (Feta cheese, raw shrimp, cooked shrimp, and cooked crawfish). However, in one Feta cheese sample and one raw shrimp sample inoculated with combined GFP-positive and GFP-negative cultures, colonies of the GFP-positive strains were not observed under UV light; fluorescing cells in one of the inoculated samples (raw shrimp) were revealed by microscopy. In general, the isolates from the inoculated foods were GFP positive by microscopic examination; the pure isolates could also be restreaked onto Trypticase soy agar, and colonies could be visually examined under UV light. Because GFP strains are not known to occur naturally in the environment, the use of the Salmonella GFP-positive strain may offer advantages as a positive control even when distinct and rare serotypes are available. The GFP-positive E. coli O157:H7 strain may also prove beneficial for use as a positive control strain for sample analyses.

COMPARATIVE TOXICITY OF FLUORANTHENE TO FRESHWATER AND SALTWATER SPECIES UNDER FLUORESCENT AND ULTRAVIOLET LIGHT

EPA Science Inventory

The acute and chronic toxicity of fluoranthene was determined for a diverse group of freshwater and saltwater species under both standard laboratory fluorescent light and ultraviolet (UV) light test conditions. Acute tests with 21 species demonstrated that fluoranthene was not le...

Two-photon excited photoconversion of cyanine-based dyes

NASA Astrophysics Data System (ADS)

Kwok, Sheldon J. J.; Choi, Myunghwan; Bhayana, Brijesh; Zhang, Xueli; Ran, Chongzhao; Yun, Seok-Hyun

2016-03-01

The advent of phototransformable fluorescent proteins has led to significant advances in optical imaging, including the unambiguous tracking of cells over large spatiotemporal scales. However, these proteins typically require activating light in the UV-blue spectrum, which limits their in vivo applicability due to poor light penetration and associated phototoxicity on cells and tissue. We report that cyanine-based, organic dyes can be efficiently photoconverted by nonlinear excitation at the near infrared (NIR) window. Photoconversion likely involves singlet-oxygen mediated photochemical cleavage, yielding blue-shifted fluorescent products. Using SYTO62, a biocompatible and cell-permeable dye, we demonstrate photoconversion in a variety of cell lines, including depth-resolved labeling of cells in 3D culture. Two-photon photoconversion of cyanine-based dyes offer several advantages over existing photoconvertible proteins, including use of minimally toxic NIR light, labeling without need for genetic intervention, rapid kinetics, remote subsurface targeting, and long persistence of photoconverted signal. These findings are expected to be useful for applications involving rapid labeling of cells deep in tissue.

Simultaneous measurements of velocity, temperature, and pressure using rapid CW wavelength-modulation laser-induced fluorescence of OH

NASA Technical Reports Server (NTRS)

Chang, A. Y.; Battles, B. E.; Hanson, R. K.

1990-01-01

In high speed flows, laser induced fluorescence (LIF) on Doppler shifted transitions is an attractive technique for velocity measurement. LIF velocimetry was applied to combined single-point measurements of velocity, temperature, and pressure and 2-D imaging of velocity and pressure. Prior to recent research using NO, LIF velocimetry in combustion related flows relied largely on the use of seed molecules. Simultaneous, single-point LIF measurements is reported of velocity, temperature, and pressure using the naturally occurring combustion species OH. This experiment is an extension of earlier research in which a modified ring dye laser was used to make time resolved temperature measurements behind reflected shock waves by using OH absorption an in postflame gases by using OH LIF. A pair of fused-silica rhombs mounted on a single galvanonmeter in an intracavity-doubled Spectra-Physics 380 ring laser permit the UV output to be swept continuously over a few wave numbers at an effective frequency of 3kHz.

Quantitative NO-LIF imaging in high-pressure flames

NASA Astrophysics Data System (ADS)

Bessler, W. G.; Schulz, C.; Lee, T.; Shin, D.-I.; Hofmann, M.; Jeffries, J. B.; Wolfrum, J.; Hanson, R. K.

2002-07-01

Planar laser-induced fluorescence (PLIF) images of NO concentration are reported in premixed laminar flames from 1-60 bar exciting the A-X(0,0) band. The influence of O2 interference and gas composition, the variation with local temperature, and the effect of laser and signal attenuation by UV light absorption are investigated. Despite choosing a NO excitation and detection scheme with minimum O2-LIF contribution, this interference produces errors of up to 25% in a slightly lean 60 bar flame. The overall dependence of the inferred NO number density with temperature in the relevant (1200-2500 K) range is low (<±15%) because different effects cancel. The attenuation of laser and signal light by combustion products CO2 and H2O is frequently neglected, yet such absorption yields errors of up to 40% in our experiment despite the small scale (8 mm flame diameter). Understanding the dynamic range for each of these corrections provides guidance to minimize errors in single shot imaging experiments at high pressure.

Comparisons Between NO PLIF Imaging and CFD Simulations of Mixing Flowfields for High-Speed Fuel Injectors

NASA Technical Reports Server (NTRS)

Drozda, Tomasz G.; Cabell, Karen F.; Ziltz, Austin R.; Hass, Neal E.; Inman, Jennifer A.; Burns, Ross A.; Bathel, Brett F.; Danehy, Paul M.

2017-01-01

The current work compares experimentally and computationally obtained nitric oxide (NO) planar laser induced fluorescence (PLIF) images of the mixing flowfields for three types of high-speed fuel injectors: a strut, a ramp, and a rectangular flushwall. These injection devices, which exhibited promising mixing performance at lower flight Mach numbers, are currently being studied as a part of the Enhanced Injection and Mixing Project (EIMP) at the NASA Langley Research Center. The EIMP aims to investigate scramjet fuel injection and mixing physics, and improve the understanding of underlying physical processes relevant to flight Mach numbers greater than eight. In the experiments, conducted in the NASA Langley Arc-Heated Scramjet Test Facility (AHSTF), the injectors are placed downstream of a Mach 6 facility nozzle, which simulates the high Mach number air flow at the entrance of a scramjet combustor. Helium is used as an inert substitute for hydrogen fuel. Both schlieren and PLIF techniques are applied to obtain mixing flowfield flow visualizations. The experimental PLIF is obtained by using a UV laser sheet to interrogate a plane of the flow by exciting fluorescence from the NO molecules, which are present in the AHSTF air. Consequently, the absence of signal in the resulting PLIF images is an indication of pure helium (fuel). The computational PLIF is obtained by applying a fluorescence model for NO to the results of the Reynolds-averaged simulations (RAS) of the mixing flow field carried out using the VULCAN-CFD solver. This approach is required because the PLIF signal is a nonlinear function of not only NO concentration, but also pressure, temperature, and the flow velocity. This complexity allows additional flow features to be identified and compared with those obtained from the computational fluid dynamics (CFD) simulations, however, such comparisons are only semiquantitative. Three-dimensional image reconstruction, similar to that used in magnetic resonance imaging, is also used to obtain images in the streamwise and spanwise planes from select cross-stream PLIF plane data. Synthetic schlieren is also computed from the RAS data. Good agreement between the experimental and computational results provides increased confidence in the CFD simulations for investigations of injector performance.

Interpretation of the fluorescence signatures from vegetation

NASA Astrophysics Data System (ADS)

Buschmann, C.

Vegetation emits fluorescence as part of the energy taken up by absorption %of solar radiation from UV to the visible. This fluorescence consists of light with low intensity (only few percents of the reflected light) emitted from the leaves. The fluorescence emission of a green leaf is characterized by four bands with maxima in the blue (440 nm), green (520 nm), red (690 nm) and far red (740 nm) spectral region. The intensity of fluorescence in the maxima of the emission spectrum varies depending on the following six basic parameters which must be taken into account for the interpretation of fluorescence signatures from vegetation: (a) content of the fluorophores (ferulic acid, chlorophyll a), (b) temperature of the leaf, (c) penetration of excitation light into the leaf, (d) emission of fluorescence from the leaf (re-absorption inside the leaf tissue), (e) photosynthetic activity of the leaf, (f) non-radiative decay (heat production) parallel to the fluorescence The ratios between the intensities of the maxima (F440/F690, F440/F520, F690/F740) are used as characteristic fluorescence parameter. The wide range of changes of these ratios caused by differences in the leaf tissue (aerial interspaces, variegated/homogeneous green leaves), various types of stress (UV, photoinhibition, sun exposure, heat, water deficiency, N-deficiency) and chemicals (inhibitors, fertilizers) can be explained by changes of the six basic parameters. It will be shown that the interpretation of the fluorescence signatures, in most cases, must be based on a complex consideration of more than one of the basic parameters.

Time-resolvable fluorescent conjugates for the detection of pathogens in environmental samples containing autofluorescent material

NASA Astrophysics Data System (ADS)

Connally, Russell; Veal, Duncan; Piper, James A.

2003-07-01

Water is routinely monitored for environmental pathogens such a Cryptosporidium and Giardia using immunofluorescence microscopy (IFM). Autofluorescence can greatly diminish an operators capacity to resolve labeled pathogens from non-specific background. Naturally fluorescing components (autofluorophores) encountered in biological samples typically have fluorescent lifetimes (τ) of less than 100 nanoseconds and their emissions may be excluded through use of time-resolved fluorescence microscopy (TRFM). TRFM relies on the large differences in τ between autofluorescent molecules and long-lived lanthanide chelates. In TRFM, targets labeled with a time-resolvable fluorescent immunoconjugate are excited by an intense (UV) light pulse. A short delay is imposed to permit the decay of autofluorescence before capture of luminescence from the excited chelate using an image intensified CCD camera. In our experience, autofluorescence can be reduced to insignificant levels with a consequent 30-fold increase in target visibility using TRFM techniques. We report conjugation of a novel europium chelate to a monoclonal antibody specific for Giardia lamblia and use of the immunoconjugate for TRFM studies. Initial attempts to conjugate the same chelate to a monoclonal antibody directed against Cryptosporidium parvum led to poorly fluorescent constructs that were prone to denature and precipitate. We successfully conjugated BHHCT to anti-mouse polyvalent immunoglobulin and used this construct to overcome the difficulties in direct labeling of the anti-Cryptosporidium antibody. Both Giardia and Cryptosporidium were labeled using the anti-mouse protocol with a subsequent 20-fold and 6.6-fold suppression of autofluorescence respectively. A rapid protocol for conjugating and purifying the immunoconjugate was found and methods of quantifying the fluorescence to protein ratio determined. Performance of our TRFM was dependent on the quality and brightness of the immunoconjugate and optimization of the conjugation process is necessary to reap the full benefit of time-resolved techniques.

Multispectral open-air intraoperative fluorescence imaging.

PubMed

Behrooz, Ali; Waterman, Peter; Vasquez, Kristine O; Meganck, Jeff; Peterson, Jeffrey D; Faqir, Ilias; Kempner, Joshua

2017-08-01

Intraoperative fluorescence imaging informs decisions regarding surgical margins by detecting and localizing signals from fluorescent reporters, labeling targets such as malignant tissues. This guidance reduces the likelihood of undetected malignant tissue remaining after resection, eliminating the need for additional treatment or surgery. The primary challenges in performing open-air intraoperative fluorescence imaging come from the weak intensity of the fluorescence signal in the presence of strong surgical and ambient illumination, and the auto-fluorescence of non-target components, such as tissue, especially in the visible spectral window (400-650 nm). In this work, a multispectral open-air fluorescence imaging system is presented for translational image-guided intraoperative applications, which overcomes these challenges. The system is capable of imaging weak fluorescence signals with nanomolar sensitivity in the presence of surgical illumination. This is done using synchronized fluorescence excitation and image acquisition with real-time background subtraction. Additionally, the system uses a liquid crystal tunable filter for acquisition of multispectral images that are used to spectrally unmix target fluorescence from non-target auto-fluorescence. Results are validated by preclinical studies on murine models and translational canine oncology models.

Detection of chitinase activity by 2-aminobenzoic acid labeling of chito-oligosaccharides.

PubMed

Ghauharali-van der Vlugt, Karen; Bussink, Anton P; Groener, Johanna E M; Boot, Rolf G; Aerts, Johannes M F G

2009-01-01

Chitinases are hydrolases capable of hydrolyzing the abundant natural polysaccharide chitin. Next to artificial fluorescent substrates, more physiological chito-oligomers are commonly used in chitinase assays. Analysis of chito-oligosaccharides products is generally accomplished by UV detection. However, the relatively poor sensitivity poses a serious limitation. Here we report on a novel, much more sensitive assay for the detection of chito-oligosaccharide reaction products released by chitinases, based on fluorescent detection, following chemical labeling by 2-aminobenzoic acid. Comparison with existing UV-based assays, shows that the novel assay offers the same advantages yet allows detection of chito-oligosaccharides in the low picomolar range.

340nm UV LED excitation in time-resolved fluorescence system for europium-based immunoassays detection

NASA Astrophysics Data System (ADS)

Rodenko, Olga; Fodgaard, Henrik; Tidemand-Lichtenberg, Peter; Pedersen, Christian

2017-02-01

In immunoassay analyzers for in-vitro diagnostics, Xenon flash lamps have been widely used as excitation light sources. Recent advancements in UV LED technology and its advantages over the flash lamps such as smaller footprint, better wall-plug efficiency, narrow emission spectrum, and no significant afterglow, have made them attractive light sources for gated detection systems. In this paper, we report on the implementation of a 340 nm UV LED based time-resolved fluorescence system based on europium chelate as a fluorescent marker. The system performance was tested with the immunoassay based on the cardiac marker, TnI. The same signal-to-noise ratio as for the flash lamp based system was obtained, operating the LED below specified maximum current. The background counts of the system and its main contributors were measured and analyzed. The background of the system of the LED based unit was improved by 39% compared to that of the Xenon flash lamp based unit, due to the LEDs narrower emission spectrum and longer pulse width. Key parameters of the LED system are discussed to further optimize the signal-to-noise ratio and signal-to-background, and hence the sensitivity of the instrument.

Remote Detection of Biological Particles and Chemical Plumes Using UV Fluorescence Lidar

NASA Technical Reports Server (NTRS)

Tiee, J. J.; Hof, D. E.; Karl, R. R.; Martinez, R. J.; Quick, C. R.; Cooper, D. I.; Eichinger, W. E.; Holtkamp, D. B.

1992-01-01

A lidar system based on ultraviolet (UV) laser induced fluorescence (LIF) was developed for the remote detection of atmospherically dispersed biological particles and chemical vapors. This UV fluorescence lidar has many potential applications for monitoring environmental pollution, industrial waste emission, agricultural insect control, illicit chemical processing, and military defense operations. The general goal of this work is to investigate the research issues associated with the long range detection and identification of chemicals, e.g. aromatic solvents and chemical precursors, and biological materials, e.g. bacillus thuringiensis (BT) and bacillus globiggi (BG). In the detection of biological particulates, we are particularly interested in extending the detection range of an existing solar-blind 248-nm lidar system. We are investigating the use of longer excitation laser wavelengths (i.e. lambda greater than 280-nm to have more favorable atmospheric light transmission characteristics) for improving detection range to better than 10 km. In the detection of chemical plumes, our main research objectives are to determine how accurately and sensitively a chemical plume can be located at range, and how well spectrally the chemical species can be measured to allow their identification.

UV light-emitting-diode photochemical mercury vapor generation for atomic fluorescence spectrometry.

PubMed

Hou, Xiaoling; Ai, Xi; Jiang, Xiaoming; Deng, Pengchi; Zheng, Chengbin; Lv, Yi

2012-02-07

A new, miniaturized and low power consumption photochemical vapor generation (PVG) technique utilizing an ultraviolet light-emitting diode (UV-LED) lamp is described, and further validated via the determination of trace mercury. In the presence of formic acid, the mercury cold vapor is favourably generated from Hg(2+) solutions by UV-LED irradiation, and then rapidly transported to an atomic fluorescence spectrometer for detection. Optimum conditions for PVG and interferences from concomitant elements were investigated in detail. Under optimum conditions, a limit of detection (LOD) of 0.01 μg L(-1) was obtained, and the precision was better than 3.2% (n = 11, RSD) at 1 μg L(-1) Hg(2+). No obvious interferences from any common ions were evident. The methodology was successfully applied to the determination of mercury in National Research Council Canada DORM-3 fish muscle tissue and several water samples.

Comparison of the effect of UV laser radiation and of a radiomimetic substance on chromatin

NASA Astrophysics Data System (ADS)

Radulescu, Irina; Radu, Liliana; Serbanescu, Ruxandra; Nelea, V. D.; Martin, C.; Mihailescu, Ion N.

1998-07-01

The damages of the complex of deoxyribonucleic acid (DNA) and proteins from chromatin, produced by the UV laser radiation and/or by treatment with a radiomimetic substance, bleomycin, were compared. The laser radiation and bleomycin effects on chromatin structure were determined by the static and dynamic fluorimetry of chromatin complexes with the DNA specific ligand-- proflavine and by the analysis of tryptophan chromatin intrinsic fluorescence. Time resolved spectroscopy is a sensitive technique which allows to determine the excited state lifetimes of chromatin--proflavine complexes. Also, the percentage contributions to the fluorescence of proflavine, bound and unbound to chromatin DNA, were evaluated. The damages produced by the UV laser radiation on chromatin are similar with those of radiomimetic substance action and consists in DNA and proteins destruction. The DNA damage degree has been determined. The obtained results may constitute some indications in the laser utilization in radiochimiotherapy.

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...

The High-Excitation Planetary Nebula NCG 246: Optical and Near-Ultraviolet Observations and Two-dimensional Numerical Models

NASA Technical Reports Server (NTRS)

Szentgyorgyi, Andrew; Raymond, John; Franco, Jose; Villaver, Eva; Lopez-Martin, Luis

2003-01-01

We have imaged the planetary nebula (PN) NGC 246 in the near-ultraviolet wavelengths [Ne v] 342.6 nm, the Bowen fluorescence line of 0 111 at 344.4 nm, and a nearby line-free region centered on 338.6 nm, as well as H(alpha), [O III] 500.7 nm, and [S II] 673.0 and 671.5 nm. Imaging in the 344.4 nm line is necessary to deconvolve contamination of the [Ne v] images by O III 342.9 nm. The emission from the shell and inner parts of the nebula is detected in [Ne v]. The radial profiles of the [Ne v] brightness decrease with radius from the exciting star, indicating that the bulk of the emission from this ion is due to the hard UV stellar radiation field, with a (probably) marginal contribution from collisional ionization in a shock between the PN shell and the interstellar medium (ISM). In contrast, the radial profiles of the emission in H(alpha), [0 III] 500.7 nm, and [S II] are flatter and peak at the location of the shell. The emission of [S II] probably traces the interaction of the PN with the ambient ISM. We also present two-dimensional numerical simulations for this PN-ISM interaction. The simulations consider the stellar motion with respect to the ambient ISM, with a velocity of 85 km/s , and include the time evolution of the wind parameters and UV radiation field from the progenitor star.

Effect of Solar Ultraviolet-B Radiation during Springtime Ozone Depletion on Photosynthesis and Biomass Production of Antarctic Vascular Plants1

PubMed Central

Xiong, Fusheng S.; Day, Thomas A.

2001-01-01

We assessed the influence of springtime solar UV-B radiation that was naturally enhanced during several days due to ozone depletion on biomass production and photosynthesis of vascular plants along the Antarctic Peninsula. Naturally growing plants of Colobanthus quitensis (Kunth) Bartl. and Deschampsia antarctica Desv. were potted and grown under filters that absorbed or transmitted most solar UV-B. Plants exposed to solar UV-B from mid-October to early January produced 11% to 22% less total, as well as above ground biomass, and 24% to 31% less total leaf area. These growth reductions did not appear to be associated with reductions in photosynthesis per se: Although rates of photosynthetic O2 evolution were reduced on a chlorophyll and a dry-mass basis, on a leaf area basis they were not affected by UV-B exposure. Leaves on plants exposed to UV-B were denser, probably thicker, and had higher concentrations of photosynthetic and UV-B absorbing pigments. We suspect that the development of thicker leaves containing more photosynthetic and screening pigments allowed these plants to maintain their photosynthetic rates per unit leaf area. Exposure to UV-B led to reductions in quantum yield of photosystem II, based on fluorescence measurements of adaxial leaf surfaces, and we suspect that UV-B impaired photosynthesis in the upper mesophyll of leaves. Because the ratio of variable to maximal fluorescence, as well as the initial slope of the photosynthetic light response, were unaffected by UV-B exposure, we suggest that impairments in photosynthesis in the upper mesophyll were associated with light-independent enzymatic, rather than photosystem II, limitations. PMID:11161031

Effect of solar ultraviolet-B radiation during springtime ozone depletion on photosynthesis and biomass production of Antarctic vascular plants.

PubMed

Xiong, F S; Day, T A

2001-02-01

We assessed the influence of springtime solar UV-B radiation that was naturally enhanced during several days due to ozone depletion on biomass production and photosynthesis of vascular plants along the Antarctic Peninsula. Naturally growing plants of Colobanthus quitensis (Kunth) Bartl. and Deschampsia antarctica Desv. were potted and grown under filters that absorbed or transmitted most solar UV-B. Plants exposed to solar UV-B from mid-October to early January produced 11% to 22% less total, as well as above ground biomass, and 24% to 31% less total leaf area. These growth reductions did not appear to be associated with reductions in photosynthesis per se: Although rates of photosynthetic O(2) evolution were reduced on a chlorophyll and a dry-mass basis, on a leaf area basis they were not affected by UV-B exposure. Leaves on plants exposed to UV-B were denser, probably thicker, and had higher concentrations of photosynthetic and UV-B absorbing pigments. We suspect that the development of thicker leaves containing more photosynthetic and screening pigments allowed these plants to maintain their photosynthetic rates per unit leaf area. Exposure to UV-B led to reductions in quantum yield of photosystem II, based on fluorescence measurements of adaxial leaf surfaces, and we suspect that UV-B impaired photosynthesis in the upper mesophyll of leaves. Because the ratio of variable to maximal fluorescence, as well as the initial slope of the photosynthetic light response, were unaffected by UV-B exposure, we suggest that impairments in photosynthesis in the upper mesophyll were associated with light-independent enzymatic, rather than photosystem II, limitations.

Light-responsive polymer microcapsules as delivery systems for natural active agents

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bizzarro, Valentina; Carfagna, Cosimo; Cerruti, Pierfrancesco

2016-05-18

In this work we report the preparation and the release behavior of UV-responsive polymeric microcapsules containing essential oils as a core. The oil acted also as a monomer solvent during polymerization. Accordingly, the potentially toxic organic solvent traditionally used was replaced with a natural active substance, resulting in a more sustainable functional system. Polymer shell was based on a lightly cross-linked polyamide containing UV-sensitive azobenzene moieties in the main chain. The micro-sized capsules were obtained via interfacial polycondensation in o/w emulsion, and their mean size was measured via Dynamic Light Scattering. Shape and morphology were analyzed through Scanning Electron andmore » Optical Microscopy. UV-responsive behavior was evaluated via spectrofluorimetry, by assessing the release kinetics of a fluorescent probe molecule upon UV light irradiation (λ{sub max}=360 nm). The irradiated samples showed an increase in fluorescence intensity, in accordance with the increase of the probe molecule concentration in the release medium. As for the un-irradiated sample, no changes could be detected demonstrating the effectiveness of the obtained releasing system.« less

Fiber-optic fluorescence imaging

PubMed Central

Flusberg, Benjamin A; Cocker, Eric D; Piyawattanametha, Wibool; Jung, Juergen C; Cheung, Eunice L M; Schnitzer, Mark J

2010-01-01

Optical fibers guide light between separate locations and enable new types of fluorescence imaging. Fiber-optic fluorescence imaging systems include portable handheld microscopes, flexible endoscopes well suited for imaging within hollow tissue cavities and microendoscopes that allow minimally invasive high-resolution imaging deep within tissue. A challenge in the creation of such devices is the design and integration of miniaturized optical and mechanical components. Until recently, fiber-based fluorescence imaging was mainly limited to epifluorescence and scanning confocal modalities. Two new classes of photonic crystal fiber facilitate ultrashort pulse delivery for fiber-optic two-photon fluorescence imaging. An upcoming generation of fluorescence imaging devices will be based on microfabricated device components. PMID:16299479

Multispectral analysis tools can increase utility of RGB color images in histology

NASA Astrophysics Data System (ADS)

Fereidouni, Farzad; Griffin, Croix; Todd, Austin; Levenson, Richard

2018-04-01

Multispectral imaging (MSI) is increasingly finding application in the study and characterization of biological specimens. However, the methods typically used come with challenges on both the acquisition and the analysis front. MSI can be slow and photon-inefficient, leading to long imaging times and possible phototoxicity and photobleaching. The resulting datasets can be large and complex, prompting the development of a number of mathematical approaches for segmentation and signal unmixing. We show that under certain circumstances, just three spectral channels provided by standard color cameras, coupled with multispectral analysis tools, including a more recent spectral phasor approach, can efficiently provide useful insights. These findings are supported with a mathematical model relating spectral bandwidth and spectral channel number to achievable spectral accuracy. The utility of 3-band RGB and MSI analysis tools are demonstrated on images acquired using brightfield and fluorescence techniques, as well as a novel microscopy approach employing UV-surface excitation. Supervised linear unmixing, automated non-negative matrix factorization and phasor analysis tools all provide useful results, with phasors generating particularly helpful spectral display plots for sample exploration.

Synthesis and crystal structure determination of copper(II)-complex: In vitro DNA and HSA binding, pBR322 plasmid cleavage, cell imaging and cytotoxic studies.

PubMed

Tabassum, Sartaj; Zaki, Mehvash; Ahmad, Musheer; Afzal, Mohd; Srivastav, Saurabh; Srikrishna, Saripella; Arjmand, Farukh

2014-08-18

New Cu(II) complex 1 of indole-3-propionic acid and 1,10-phenanthroline was synthesized and characterized by analytical, spectroscopic and single crystal X-ray diffraction. In vitro DNA binding studies of 1 was performed by employing UV-vis and fluorescence spectroscopic techniques. The binding affinity towards human serum albumin (HSA) was also investigated to understand the carrier role in body system, as the time dependent HPLC experiment of 1 revealed that bonded drug with protein releases slowly in presence of DNA. Complex 1 exhibited good anti-tumor activity (GI50 values <10 μg/ml), and to elucidate the mechanism of tumor inhibition, topoisomerase I enzymatic activity was carried out and further validated by cell imaging studies which clearly showed its nuclear localization. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Identification of forensic samples by using an infrared-based automatic DNA sequencer.

PubMed

Ricci, Ugo; Sani, Ilaria; Klintschar, Michael; Cerri, Nicoletta; De Ferrari, Francesco; Giovannucci Uzielli, Maria Luisa

2003-06-01

We have recently introduced a new protocol for analyzing all core loci of the Federal Bureau of Investigation's (FBI) Combined DNA Index System (CODIS) with an infrared (IR) automatic DNA sequencer (LI-COR 4200). The amplicons were labeled with forward oligonucleotide primers, covalently linked to a new infrared fluorescent molecule (IRDye 800). The alleles were displayed as familiar autoradiogram-like images with real-time detection. This protocol was employed for paternity testing, population studies, and identification of degraded forensic samples. We extensively analyzed some simulated forensic samples and mixed stains (blood, semen, saliva, bones, and fixed archival embedded tissues), comparing the results with donor samples. Sensitivity studies were also performed for the four multiplex systems. Our results show the efficiency, reliability, and accuracy of the IR system for the analysis of forensic samples. We also compared the efficiency of the multiplex protocol with ultraviolet (UV) technology. Paternity tests, undegraded DNA samples, and real forensic samples were analyzed with this approach based on IR technology and with UV-based automatic sequencers in combination with commercially-available kits. The comparability of the results with the widespread UV methods suggests that it is possible to exchange data between laboratories using the same core group of markers but different primer sets and detection methods.

Investigation on the pH-dependent binding of benzocaine and lysozyme by fluorescence and absorbance

NASA Astrophysics Data System (ADS)

Li, Shihui; Li, Daojin

2011-11-01

The interaction mechanism between benzocaine (BZC) and lysozyme (Lys) has been investigated by fluorescence, synchronous fluorescence, ultraviolet-vis (UV) absorption spectra, and three-dimensional fluorescence (3-D) in various pH medium. The observations of fluorescence spectra were mainly rationalized in terms of a static quenching process at lower concentration of BZC ( CBZC/ CLys < 9) and a combined quenching process at higher concentration of BZC ( CBZC/ CLys > 9) at pH 7.4 and 8.4. However, the fluorescence quenching was mainly arisen from static quenching by complex formation in all studied drug concentrations at pH 3.5. The structural characteristics of BZC and Lys were probed, and their binding affinities were determined under different pH conditions (pH 3.5, 7.4, and 8.4). The results indicated that the binding abilities of BZC to Lys decreased at the pH below and above the simulative physiological condition (pH 7.4) due to the alterations of the protein secondary and tertiary structures or the structural change of BZC. The effect of BZC on the conformation of Lys was analyzed using UV, synchronous fluorescence and three-dimensional fluorescence under different pH conditions. These results indicate that the binding of BZC to Lys causes apparent change in the secondary and tertiary structures of Lys. The effect of Zn 2+ on the binding constant of BZC with Lys under various pH conditions (pH 3.5, 7.4, and 8.4) was also studied.

Two sugar-rhodamine "turn-on" fluorescent probes for the selective detection of Fe3 +

NASA Astrophysics Data System (ADS)

Chen, Qing; Fang, Zhijie

2018-03-01

Two new sugar-rhodamine fluorescent probes (RDG1 and RDG2) have been synthesized and characterized by 1H NMR, 13C NMR and HRMS. Their UV-Vis, fluorescence spectra and fluorescence-response to Fe3 + are investigated and discussed. RDG1 had a very nice linear relationship between UV absorbance and Fe3 + concentration with the correlation coefficient as high as 0.997 and the detection limit is 3.46 × 10- 6 M. Upon the addition of Fe3 +, the spirolactam ring of RDG1 was opened and a 1:1 metal ligand complex was formed from Job's plot. The results showed that RDG1 can be used as an effective fluorescent probe for selective detection of Fe3 + in water. RDG2 was incorporated the well-known rhodamine group and a water-soluble D-glucose group within one molecule and can be used for detecting Fe3 + in natural water as a selective fluorescent sensor. The addition of Fe3 + into RDG2 resulted in a strongly enhanced fluorescence as well as color change of solution from colorless to pink. Job's plot of RDG2 indicated 1:1 stoichiometry of RDG2-Fe3 +. RDG2 can serve as a probe for Fe3 + between pH = 4.0 to 7.0 and it's detection limit is 2.09 × 10- 6 M. The OFF-ON fluorescent mechanisms of RDG1-Fe3 + and RDG2-Fe3 + are proposed.

Monitoring biological aerosols using UV fluorescence

NASA Astrophysics Data System (ADS)

Eversole, Jay D.; Roselle, Dominick; Seaver, Mark E.

1999-01-01

An apparatus has been designed and constructed to continuously monitor the number density, size, and fluorescent emission of ambient aerosol particles. The application of fluorescence to biological particles suspended in the atmosphere requires laser excitation in the UV spectral region. In this study, a Nd:YAG laser is quadrupled to provide a 266 nm wavelength to excite emission from single micrometer-sized particles in air. Fluorescent emission is used to continuously identify aerosol particles of biological origin. For calibration, biological samples of Bacillus subtilis spores and vegetative cells, Esherichia coli, Bacillus thuringiensis and Erwinia herbicola vegetative cells were prepared as suspensions in water and nebulized to produce aerosols. Detection of single aerosol particles, provides elastic scattering response as well as fluorescent emission in two spectral bands simultaneously. Our efforts have focuses on empirical characterization of the emission and scattering characteristics of various bacterial samples to determine the feasibility of optical discrimination between different cell types. Preliminary spectroscopic evidence suggest that different samples can be distinguished as separate bio-aerosol groups. In addition to controlled sample results, we will also discuss the most recent result on the effectiveness of detection outdoor releases and variations in environmental backgrounds.

Non-covalent attachment of silver nanoclusters onto single-walled carbon nanotubes with human serum albumin as linking molecule

NASA Astrophysics Data System (ADS)

Rodríguez-Galván, Andrés; Heredia, Alejandro; Amelines-Sarria, Oscar; Rivera, Margarita; Medina, Luis A.; Basiuk, Vladimir A.

2015-03-01

The attachment of silver nanoclusters (AgNCs) onto single-walled carbon nanotubes (SWNTs) for the formation of integrated fluorescence sites has attracted much attention due their potential applications as biological probes and nanovectors in theragnosis. Here, we report the preparation through assembly of fluorescent quasi 1-D nanomaterial based on SWNTs and silver nanoclusters (AgNCs) non-covalently attached to human serum albumin as biological linker. The fluorescent SWNT-AgNCs-HSA conjugates were characterized by atomic force microscopy, high-resolution transmission electron microscopy (HRTEM), high angle annular dark field scanning TEM (HAADF-STEM), fluorescent and UV-vis spectroscopy. The above techniques confirmed that AgNCs were non-covalently attached onto the external surface of SWNTs. In addition, it was observed that the modification did not affect the optical properties of the synthesized AgNCs since the absorption spectra and fluorescence under UV irradiation (λ = 365 nm) remain the same. The effect of the functionalized systems was tested on mammal red blood cells (RBCs) and it was found that their structural integrity was compromised by the conjugates, limiting their biological and medical applications.

UV/PAR radiation and DOM properties in surface coastal waters of the Canadian shelf of the Beaufort Sea during summer 2009

NASA Astrophysics Data System (ADS)

Para, J.; Charrière, B.; Matsuoka, A.; Miller, W. L.; Rontani, J. F.; Sempéré, R.

2013-04-01

Surface waters from the Beaufort Sea in the Arctic Ocean were evaluated for dissolved organic carbon (DOC), and optical characteristics including UV (ultraviolet) radiation and PAR (photosynthetically active radiation) diffuse attenuation (Kd), and chromophoric and fluorescent dissolved organic matter (CDOM and FDOM) as part of the MALINA field campaign (30 July to 27 August). Spectral absorption coefficients (aCDOM (350 nm) (m-1)) were significantly correlated to both diffuse attenuation coefficients (Kd) in the UV-A and UV-B and to DOC concentrations. This indicates CDOM as the dominant attenuator of both UV and PAR solar radiation and suggests its use as an optical proxy for DOC concentrations in this region. While the Mackenzie input is the main driver of CDOM dynamics in low salinity waters, locally, primary production can create significant increases in CDOM. Extrapolating CDOM to DOC relationships, we estimate that ∼16% of the DOC in the Mackenzie River does not absorb radiation at 350 nm. The discharges of DOC and its chromophoric subset (CDOM) by the Mackenzie River during the MALINA cruise are estimated as ∼0.22 TgC and 0.18 TgC, respectively. Three dissolved fluorescent components (C1-C3) were identified by fluorescence excitation/emission matrix spectroscopy (EEMS) and parallel factor (PARAFAC) analysis. Our results showed an aquatic dissolved organic matter (DOM) component (C1), probably produced in the numerous lakes of the watershed, that co-dominated with a terrestrial humic-like component (C2) in the Mackenzie Delta Sector. This aquatic DOM could partially explain the high CDOM spectral slopes observed in the Beaufort Sea.

Durability of Polymeric Encapsulation Materials for a PMMA/glass Concentrator Photovoltaic System

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, David C.; Kempe, Michael D.; Muller, Matthew T

2014-04-08

The durability of polymeric encapsulation materials was examined using outdoor exposure at the nominal geometric concentration of 500 suns. The results for 36 months cumulative field deployment are presented for materials including: poly(ethylene-co-vinyl acetate), (EVA); polyvinyl butyral (PVB); ionomer; polyethylene/ polyoctene copolymer (PO); thermoplastic polyurethane (TPU); poly(dimethylsiloxane) (PDMS); poly(diphenyl dimethyl siloxane) (PDPDMS); and poly(phenyl-methyl siloxane) (PPMS). Measurements of the field conditions including ambient temperature and ultraviolet (UV) dose were recorded at the test site during the experiment. Measurements for the experiment included optical transmittance (with subsequent analysis of solar-weighted transmittance, UV cut-off wavelength, and yellowness index), mass, visual photography, photoelasticmore » imaging, and fluorescence spectroscopy. While the results to date for EVA are presented and discussed, examination here focuses more on the siloxane materials. A specimen recently observed to fail by thermal decomposition is discussed in terms of the implementation of the experiment as well as its fluorescence signature, which was observed to become more pronounced with age. Modulated thermogravimetry (allowing determination of the activation energy of thermal decomposition) was performed on a subset of the siloxanes to quantify the propensity for decomposition at elevated temperatures. Supplemental, Pt-catalyst- and primer-solutions as well as peroxide-cured PDMS specimens were examined to assess the source of the luminescence. The results of the study including the change in optical transmittance, observed failure modes, and subsequent analyses of the failure modes are described in the conclusions.« less

Durability of polymeric encapsulation materials in a PMMA/glass concentrator photovoltaic system

DOE PAGES

Miller, David C.; Kempe, Michael D.; Muller, Matthew T.; ...

2016-07-13

We examined the durability of polymeric encapsulation materials using outdoor exposure at the nominal geometric concentration of 500 suns. The results for 36-month cumulative field deployment are presented for materials including: poly(ethylene-co-vinyl acetate), (EVA); polyvinyl butyral (PVB); ionomer; polyethylene/polyoctene copolymer (PO); thermoplastic polyurethane (TPU); poly(dimethylsiloxane) (PDMS); poly(diphenyl dimethyl siloxane) (PDPDMS); and poly(phenyl-methyl siloxane) (PPMS). Measurements of the field conditions including ambient temperature and ultraviolet (UV) dose were recorded at the test site during the experiment. Our measurements for the experiment included optical transmittance (with subsequent analysis of solar-weighted transmittance, UV cut-off wavelength, and yellowness index), mass, visual photography, photoelastic imaging,more » and fluorescence spectroscopy. While the results to date for EVA are presented and discussed, examination here focuses more on the siloxane materials. A specimen recently observed to fail by thermal decomposition is discussed in terms of the implementation of the experiment as well as its fluorescence signature, which was observed to become more pronounced with age. Modulated thermogravimetry (allowing determination of the activation energy of thermal decomposition) was performed on a subset of the siloxanes to quantify the propensity for decomposition at elevated temperatures. Supplemental, Pt-catalyst- and primer-solutions as well as peroxide-cured PDMS specimens were examined to assess the source of the luminescence. Furthermore, our results, including the change in optical transmittance, observed failure modes, and subsequent analyses of the failure modes are described in the conclusions.« less

Hyperspectral small animal fluorescence imaging: spectral selection imaging

NASA Astrophysics Data System (ADS)

Leavesley, Silas; Jiang, Yanan; Patsekin, Valery; Hall, Heidi; Vizard, Douglas; Robinson, J. Paul

2008-02-01

Molecular imaging is a rapidly growing area of research, fueled by needs in pharmaceutical drug-development for methods for high-throughput screening, pre-clinical and clinical screening for visualizing tumor growth and drug targeting, and a growing number of applications in the molecular biology fields. Small animal fluorescence imaging employs fluorescent probes to target molecular events in vivo, with a large number of molecular targeting probes readily available. The ease at which new targeting compounds can be developed, the short acquisition times, and the low cost (compared to microCT, MRI, or PET) makes fluorescence imaging attractive. However, small animal fluorescence imaging suffers from high optical scattering, absorption, and autofluorescence. Much of these problems can be overcome through multispectral imaging techniques, which collect images at different fluorescence emission wavelengths, followed by analysis, classification, and spectral deconvolution methods to isolate signals from fluorescence emission. We present an alternative to the current method, using hyperspectral excitation scanning (spectral selection imaging), a technique that allows excitation at any wavelength in the visible and near-infrared wavelength range. In many cases, excitation imaging may be more effective at identifying specific fluorescence signals because of the higher complexity of the fluorophore excitation spectrum. Because the excitation is filtered and not the emission, the resolution limit and image shift imposed by acousto-optic tunable filters have no effect on imager performance. We will discuss design of the imager, optimizing the imager for use in small animal fluorescence imaging, and application of spectral analysis and classification methods for identifying specific fluorescence signals.

Probing the binding interaction of a phenazinium dye with serum transport proteins: a combined fluorometric and circular dichroism study.

PubMed

Bose, Debosreeta; Sarkar, Deboleena; Chattopadhyay, Nitin

2010-01-01

In the present investigation, an attempt has been made to study the interaction of phenosafranin (PSF), a cationic phenazinium dye with the transport proteins, bovine serum albumin (BSA) and human serum albumin (HSA), employing steady-state and time-resolved fluorometric and circular dichroism (CD) techniques. The photophysical properties of the dye are altered on binding with the serum proteins. An explicit study with respect to the modification of the fluorescence and fluorescence anisotropy upon binding, effect of denaturant, fluorescence lifetime and CD measurements reveal that the dye binds to both BSA and HSA with almost the same affinity. Far-UV CD spectra indicate a decrease in the percentage of alpha-helicity only for BSA upon binding with the probe. Near-UV CD responses indicate an alteration in the tertiary structure of both the transport proteins because of binding.