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1

Development of Pathological Diagnostics of Human Kidney Cancer by Multiple Staining Using New Fluorescent Fluolid Dyes  

PubMed Central

New fluorescent Fluolid dyes have advantages over others such as stability against heat, dryness, and excess light. Here, we performed simultaneous immunostaining of renal tumors, clear cell renal cell carcinoma (RCC), papillary RCC, chromophobe RCC, acquired cystic disease-associated RCC (ACD-RCC), and renal angiomyolipoma (AML), with primary antibodies against Kank1, cytokeratin 7 (CK7), and CD10, which were detected with secondary antibodies labeled with Fluolid-Orange, Fluolid-Green, and Alexa Fluor 647, respectively. Kank1 was stained in normal renal tubules, papillary RCC, and ACD-RCC, and weakly or negatively in all other tumors. CK7 was positive in normal renal tubules, papillary RCC, and ACD-RCC. In contrast, CD10 was expressed in renal tubules and clear cell RCC, papillary RCC, AML, and AC-RCC, and weakly in chromophobe RCC. These results may contribute to differentiating renal tumors and subtypes of RCCs. We also examined the stability of fluorescence and found that fluorescent images of Fluolid dyes were identical between a tissue section and the same section after it was stored for almost three years at room temperature. This indicates that tissue sections can be stored at room temperature for a relatively long time after they are stained with multiple fluorescent markers, which could open a door for pathological diagnostics. PMID:24995295

Wuxiuer, Dilibaier; Zhu, Yun; Ogaeri, Takunori; Mizuki, Keiji; Kashiwa, Yuki; Nishi, Kentaro; Isobe, Shin-ichiro; Aoyagi, Tei-ichiro; Kiyama, Ryoiti

2014-01-01

2

CD133(+) Human Colorectal Adenocarcinoma Cells Are Resistant to Staining with Fluorescent Dyes Used for Analysis of ABC Transporter Activities.  

PubMed

Flow cytometry measurement of the expression of surface marker CD133 simultaneously with the analysis of fluorescent dye exclusion was performed in order to develop new methods for detection of cancer stem cell populations in tumor tissue samples from patients with colorectal adenocarcinoma. No correlation was found between the count of CD133(+) cancer cells and the volume of the "population" formed from cells actively pumping off the fluorescent dye. On the other hand, the fluorescence distribution plot showed predominant location of CD133(+) cancer cells among cells stained with neither DyeCycle Violet DNA-binding dye, nor rhodamine 123 mitochondrial dye. These cells did not show the properties of the classical "side population", because they did not shift to the area of stained cell after treatment with ionic channel blocker verapamil. PMID:25403403

Gisina, A M; Lupatov, A Yu; Karalkin, P A; Mainovskaya, O A; Petrov, L O; Sidorov, D V; Frank, G A; Yarygin, K N

2014-11-01

3

A fast and cost-effective methodology for Fonsecaea pedrosoi ATCC46428 staining using ESIPT fluorescent dyes.  

PubMed

The microscopic morphology of Fonsecaea pedrosoi ATCC46428 was observed using two benzazole derivatives, 2-(2'-hydroxyphenyl)benzoxazole and 2-(5'-amino-2'-hydroxyphenyl)benzoxazole, which emit intense fluorescence by a proton transfer mechanism in the electronically excited state (ESIPT). The cell surface could be successfully stained with fluorescent dye solutions of 10 microM-10 mM using two different fast and cost-effective procedures. At these concentrations, any structure or dye crystallization could be observed. Concerning the external microstructural details, only the amino derivative allowed the differentiation between hyphae and conidia. These dyes presented some advantages comparing to commercial dyes, since the stained cells showed high chemical, thermal and photochemical stability during the experiments and also after several months of storage at room temperature and normal light exposition. Procedure 1 presented the advantage to be used when heating can change the chemical or biochemical cell composition. On the other hand Procedure 2 showed to be useful as a routine methodology for cells staining. The results allowed to propose a simple and highly sensitive assay to study the F. pedrosoi micromorphology by epifluorescence microscopy. This methodology can probably be extended for other fungi of clinical interest. PMID:20385502

Corbellini, Valeriano Antonio; Scroferneker, Maria Lúcia; Carissimi, Mariana; Rodembusch, Fabiano Severo; Stefani, Valter

2010-06-01

4

A Novel Staining Protocol for Multiparameter Assessment of Cell Heterogeneity in Phormidium Populations (Cyanobacteria) Employing Fluorescent Dyes  

PubMed Central

Bacterial populations display high heterogeneity in viability and physiological activity at the single-cell level, especially under stressful conditions. We demonstrate a novel staining protocol for multiparameter assessment of individual cells in physiologically heterogeneous populations of cyanobacteria. The protocol employs fluorescent probes, i.e., redox dye 5-cyano-2,3-ditolyl tetrazolium chloride, ‘dead cell’ nucleic acid stain SYTOX Green, and DNA-specific fluorochrome 4?,6-diamidino-2-phenylindole, combined with microscopy image analysis. Our method allows simultaneous estimates of cellular respiration activity, membrane and nucleoid integrity, and allows the detection of photosynthetic pigments fluorescence along with morphological observations. The staining protocol has been adjusted for, both, laboratory and natural populations of the genus Phormidium (Oscillatoriales), and tested on 4 field-collected samples and 12 laboratory strains of cyanobacteria. Based on the mentioned cellular functions we suggest classification of cells in cyanobacterial populations into four categories: (i) active and intact; (ii) injured but active; (iii) metabolically inactive but intact; (iv) inactive and injured, or dead. PMID:23437052

Tashyreva, Daria; Elster, Josef; Billi, Daniela

2013-01-01

5

Methods of staining and visualization of sphingolipid enriched and non-enriched plasma membrane regions of Arabidopsis thaliana with fluorescent dyes and lipid analogues  

PubMed Central

Background Sterols and Sphingolipids form lipid clusters in the plasma membranes of cell types throughout the animal and plant kingdoms. These lipid domains provide a medium for protein signaling complexes at the plasma membrane and are also observed to be principal regions of membrane contact at the inception of infection. We visualized different specific fluorescent lipophilic stains of the both sphingolipid enriched and non-sphingolipid enriched regions in the plasma membranes of live protoplasts of Arabidopsis thaliana. Results Lipid staining protocols for several fluorescent lipid analogues in plants are presented. The most emphasis was placed on successful protocols for the single and dual staining of sphingolipid enriched regions and exclusion of sphingolipid enriched regions on the plasma membrane of Arabidopsis thaliana protoplasts. A secondary focus was placed to ensure that these staining protocols presented still maintain cell viability. Furthermore, the protocols were successfully tested with the spectrally sensitive dye Laurdan. Conclusion Almost all existing staining procedures of the plasma membrane with fluorescent lipid analogues are specified for animal cells and tissues. In order to develop lipid staining protocols for plants, procedures were established with critical steps for the plasma membrane staining of Arabidopsis leaf tissue and protoplasts. The success of the plasma membrane staining protocols was additionally verified by measurements of lipid dynamics by the fluorescence recovery after photobleaching technique and by the observation of new phenomena such as time dependent lipid polarization events in living protoplasts, for which a putative physiological relevance is suggested. PMID:22867517

2012-01-01

6

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

PubMed

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

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

2014-12-01

7

Exploring the dynamics of fluorescence staining of bacteria with cyanine dyes for the development of kinetic assays  

NASA Astrophysics Data System (ADS)

Bacterial infections continue to be one of the major health risks in the United States. The common occurrence of such infection is one of the major contributors to the high cost of health care and significant patient mortality. The work presented in this thesis describes spectroscopic studies that will contribute to the development of a fluorescent assay that may allow the rapid identification of bacterial species. Herein, the optical interactions between six bacterial species and a series of thiacyanine dyes are investigated. The interactions between the dyes and the bacterial species are hypothesized to be species-specific. For this thesis, two Gram-negative strains, Escherichia coli (E. coli) TOP10 and Enterobacter aerogenes; two Gram-positive bacterial strains, Bacillus sphaericus and Bacillus subtilis; and two Bacillus endospores, B. globigii and B. thuringiensis, were used to test the proposed hypothesis. A series of three thiacyanine dyes---3,3'-diethylthiacyanine iodide (THIA), 3,3'-diethylthiacarbocyanine iodide (THC) and thiazole orange (THO)---were used as fluorescent probes. The basis of our spectroscopic study was to explore the bacterium-induced interactions of the bacterial cells with the individual thiacyanine dyes or with a mixture of the three dyes. Steady-state absorption spectroscopy revealed that the different bacterial species altered the absorption properties of the dyes. Mixed-dye solutions gave unique absorption patterns for each bacteria tested, with competitive binding observed between the bacteria and spectrophotometric probes (thiacyanine dyes). Emission spectroscopy recorded changes in the emission spectra of THIA following the introduction of bacterial cells. Experimental results revealed that the emission enhancement of the dyes resulted from increases in the emission quantum yield of the thiacyanine dyes upon binding to the bacteria cellular components. The recorded emission enhancement data were fitted to an exponential (mono-exponential or bi-exponential) function, and time constants were extracted by regressing on the experimental data. The addition of the TWEEN surfactants decreased the rate at which the dyes interacted with the bacterial cells, which typically resulted in larger time constants derived from an exponential fit. ANOVA analysis of the time constants confirmed that the values of the time constants clustered in a narrow range and were independent of dye concentration and weakly dependent on cell density.

Thomas, Marlon Sheldon

8

Detection of glycoproteins in polyacrylamide gels and on electroblots using Pro-Q Emerald 488 dye, a fluorescent periodate Schiff-base stain.  

PubMed

Pro-Q Emerald 488 glycoprotein stain reacts with periodic acid-oxidized carbohydrate groups, generating a bright green-fluorescent signal on glycoproteins. The stain permits detection of less than 5-18 ng of glycoprotein per band, depending upon the nature and the degree of protein glycosylation, making it roughly 8-16-fold more sensitive than the standard colorimetric periodic acid-Schiff base method using acidic fuchsin dye (pararosaniline). The green-fluorescent signal from Pro-Q Emerald 488 stain may optimally be visualized using charge-coupled device/xenon arc lamp-based imaging systems or 470-488 nm laser-based gel scanners. Though glycoprotein detection may be performed on transfer membranes, direct detection in gels avoids electroblotting and the specificity of staining is better in gels. After detecting glycoproteins with Pro-Q Emerald 488 dye, total protein profiles may subsequently be detected using SYPRO Ruby protein gel stain. Using computer-assisted registration techniques, images may then be merged to generate differential display maps. PMID:12601726

Hart, Courtenay; Schulenberg, Birte; Steinberg, Thomas H; Leung, Wai-Yee; Patton, Wayne F

2003-02-01

9

Pyronin Y as a Fluorescent Stain for Paraffin Sections  

Microsoft Academic Search

Pyronin Y has long been used, in combination with other dyes such as Methyl Green, as a differential stain for nucleic acids\\u000a in paraffin tissue sections. It also forms fluorescent complexes with double-stranded nucleic acids, especially RNA, enabling\\u000a semi-quantitative analysis of cellular RNA in flow cytometry. However, the possibility of using pyronin Y as a fluorescent\\u000a stain for paraffin tissue

Bo Li; Ying Wu; Xiao-Ming Gao

2002-01-01

10

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2011 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2011-04-01

11

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2012 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2012-04-01

12

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2013 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2013-04-01

13

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2014 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2014-04-01

14

Fiberized fluorescent dye microtubes  

NASA Astrophysics Data System (ADS)

In the present work we study the effect of the length of fluorescent dye-filled micro-capillaries on the fluorescence spectra. Two types of micro-capillaries have been studied: a 100 ?m inner diameter fused silica capillary with a transparent coating and one of the holes of a fiber optic glass ferrule with 125 ?m inner diameter. The tubes were filled with solutions of Rhodamine 6G dissolved in ethanol and then in glycerin. Experimental data show that the maximum fluorescence and the largest spectral widths are observed for a sample length of about 0.25 mm for the used concentration. This results show that miniature tunable fiberized dye lasers can be developed using available standard micro-and fibre-optic components.

Vladev, Veselin; Eftimov, Tinko

2013-03-01

15

Nile red: a selective fluorescent stain for intracellular lipid droplets  

Microsoft Academic Search

We report that the dye nile red, 9-diethylamino-5H-benzo(a)phenoxazine-5-one, is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. The specificity of the dye for lipid droplets was assessed on cultured aortic smooth muscle cells and on cultured peritoneal macrophages that were incubated with acetylated low density lipoprotein to induce cytoplasmic lipid overloading.

PHILLIP GREENSPAN; EUGENE P. MAYER; STANLEY D. FOWLER

1985-01-01

16

A fluorescent Gram stain for flow cytometry and epifluorescence microscopy.  

PubMed

The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by SYTO 13; conversely, gram-positive organisms were rendered red-orange fluorescent by HI, which simultaneously quenched SYTO 13 green fluorescence. The technique correctly predicted the Gram status of 45 strains of clinically relevant organisms, including several known to be gram variable. In addition, representative strains of gram-positive anaerobic organisms, normally decolorized during the traditional Gram stain procedure, were classified correctly by this method. PMID:9647848

Mason, D J; Shanmuganathan, S; Mortimer, F C; Gant, V A

1998-07-01

17

Fluorescence changes during electrical activity in frog muscle stained with merocyanine  

Microsoft Academic Search

NERVE membranes stained with various fluorescent dyes show changes in fluorescence intensity concomitant with changes in transmembrane potential1-4. Cohen et al.4 described a number of dyes that exhibit changes in fluorescence emission which vary linearly with transmembrane potential. Two of these have been used to investigate the excitation-contraction process in skeletal muscle5-7. Whole muscles and single fibres stained with Nile

Julio Vergara; Francisco Bezanilla

1976-01-01

18

Improving acid-fast fluorescent staining for the detection of mycobacteria using a new nucleic acid staining approach.  

PubMed

Acid fast staining of sputum smears by microscopy remains the prevalent method for detecting Mycobacterium tuberculosis. The sensitivity of microscopy using acid fast stains requires 10(4) bacilli per ml of sputum. Although fluorescent acid fast stains, such as Auramine-O, show improved sensitivity, almost half of culture-positive TB cases are currently estimated to remain smear-negative. These current diagnosis problems provide impetus for improving staining procedures. We evaluated a novel fluorescent acid-fast staining approach using the nucleic acid-binding dye SYBR(®) Gold on mycobacterial in vitro cultures. The SYBR(®) Gold stain detected 99% of MTB in both actively replicating aerobic and non-replicating hypoxic cultures. Transmission light microscopy with Ziehl-Neelsen fuchsin, and fluorescence microscopy with Auramine-O or Auramine-rhodamine detected only 54%-86% of MTB bacilli. SYBR(®) Gold fluoresces more intensely than Auramine-O, and is highly resistant to fading. The signal to noise ratio is exceptionally high due to a >1000-fold enhanced fluorescence after binding to DNA/RNA, thereby reducing most background fluorescence. Although cost and stability of the dye may perhaps limit its clinical use at this time, these results warrant further research into more nucleic acid dye variants. In the meantime, SYBR(®) Gold staining shows great promise for use in numerous research applications. PMID:25130623

Ryan, Gavin J; Shapiro, Howard M; Lenaerts, Anne J

2014-09-01

19

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

21 Food and Drugs 8 2010-04-01 2010-04-01... 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT... HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

2010-04-01

20

Storable, thermally activated, near-infrared chemiluminescent dyes and dye-stained microparticles for optical imaging  

PubMed Central

Optical molecular imaging employs relatively harmless, low-energy light and technically straightforward instrumentation. Self-illuminating, chemiluminescent systems are especially attractive since they have inherently high signal contrast due to the lack of background emission. Currently, chemiluminescence imaging involves short-lived molecular species that are not stored but instead generated in situ, and they typically emit visible light, which does not penetrate far through heterogeneous biological media. Here, we describe a new paradigm for optical molecular imaging using squaraine rotaxane endoperoxides (SREPs), interlocked fluorescent and chemiluminescent dye molecules that have a squaraine chromophore encapsulated inside a macrocycle endoperoxide. SREPs can be stored indefinitely at temperatures below ?20 °C, but upon warming to body temperature they undergo a unimolecular chemical reaction and emit near infrared light that can pass through a living mouse. Dye-stained microparticles are easily prepared for in vivo near-infrared optical imaging using commercial imaging stations. PMID:21107365

Baumes, Jeffrey M.; Gassensmith, Jeremiah J.; Giblin, Jay; Lee, Jung-Jae; White, Alexander G.; Culligan, William J.; Leevy, W. Matthew; Kuno, Masaru; Smith, Bradley D.

2011-01-01

21

Investigating fluorescent dyes in fluorescence-assisted screenings.  

PubMed

Screening of bead-based peptide libraries against fluorescent dye-labeled target proteins was found to be significantly influenced by the dye characteristics. Commercially available red fluorescent dyes with net negative charges adversely showed strong interactions with library beads. The introduction of zwitterionic dyes significantly reduced the unwanted interactions, which sheds light upon using the right fluorescent probe for acquisition of reliable results in various fluorescence-assisted applications. PMID:25340456

Jee, Joo-Eun; Lim, Jaehong; Hyun, Hoon; Oon, Jessica; Ong, Yong Siang; Massif, Cedrik; Chang, Young-Tae; Choi, Hak Soo; Lee, Su Seong

2014-12-14

22

The Polarization of Fluorescence of DNA Stains Depends on the Incorporation Density of the  

E-print Network

The Polarization of Fluorescence of DNA Stains Depends on the Incorporation Density of the Dye: The fluorescence induced by polarized light sources, such as the lasers that are used in flow cytometry, is often polarized and anisotropic. In addition, most optical detector systems are sensitive to the direc- tion

Asbury, Chip

23

Dimeric fluorescent energy transfer dyes comprising asymmetric cyanine azole-indolenine chromophores  

DOEpatents

Novel fluorescent DNA-staining dyes are provided combining asymmetric cyanine azole-indolenine dyes, which provide for strong DNA affinity, large Stokes shifts and emission in the red region of the spectrum. The dyes find particular application in gel electrophoresis and for labels which may be bound to a variety of compositions in a variety of contexts.

Glazer, Alexander N. (Orinda, CA); Benson, Scott C. (Albany, CA)

1996-01-01

24

Classification and naming of dyes, stains and fluorochromes.  

PubMed

A classification of dyes and other colorants is proposed, based on the chemical features responsible for their visibility and generally consonant with the writings of modern color chemists. The scheme differs in several respects from that of the Colour Index (CI), but it retains some traditional small groups of dyes that include biological stains. Natural dyes, recognized as a group in the CI, are placed with or near synthetic dyes with identical or similar chromophores. The new scheme also provides categories for dyes and fluorochromes that do not have places in the CI classification. Some CI categories, including lactones, aminoketones and hydroxyketones, are not recognized in this new scheme, which is adopted in the forthcoming 10th edition of Conn's Biological Stains: a Handbook of Dyes and Fluorochromes for Use in Biology and Medicine. Some rules are also set out for the spelling of trivial names, which has long been inconsistent in scientific literature. The ending '-ine' is used for compounds derived from organic bases (e.g., fuchsine and thionine, not fuchsin or thionin), and names ending in '-in' are for compounds that are not bases or their derivatives (e.g., eosin and phloxin, not eosine or phloxine). Initial capital letters are used only for words that are names of people or places (e.g., Nile blue or Congo red) and for the 'generic' components of CI application names (as in Acid yellow 36). Other words, including trade names that have fallen into common usage are not capitalized (e.g., alcian blue, biebrich scarlet, coomassie blue). The recommended spellings of some dyes differ from those commonly seen in vendors' catalogs and in biological publications, but they are generally consistent with English and American dictionaries, with recent writings in English by color chemists, and with the trivial names of other organic compounds. PMID:11871748

Kiernan, J A

2001-01-01

25

Specific in vivo staining of astrocytes in the whole brain after intravenous injection of sulforhodamine dyes.  

PubMed

Fluorescent staining of astrocytes without damaging or interfering with normal brain functions is essential for intravital microscopy studies. Current methods involved either transgenic mice or local intracerebral injection of sulforhodamine 101. Transgenic rat models rarely exist, and in mice, a backcross with GFAP transgenic mice may be difficult. Local injections of fluorescent dyes are invasive. Here, we propose a non-invasive, specific and ubiquitous method to stain astrocytes in vivo. This method is based on iv injection of sulforhodamine dyes and is applicable on rats and mice from postnatal age to adulthood. The astrocytes staining obtained after iv injection was maintained for nearly half a day and showed no adverse reaction on astrocytic calcium signals or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows to quantify 3D morphological parameters of the astrocytes and to characterize their network. Our method may become a reference for in vivo staining of the whole astrocytes population in animal models of neurological disorders. PMID:22509398

Appaix, Florence; Girod, Sabine; Boisseau, Sylvie; Römer, Johannes; Vial, Jean-Claude; Albrieux, Mireille; Maurin, Mathieu; Depaulis, Antoine; Guillemain, Isabelle; van der Sanden, Boudewijn

2012-01-01

26

Reactive Fluorescent Dyes For Urethane Coatings  

NASA Technical Reports Server (NTRS)

Molecules of fluorescent dyes chemically bound in urethane conformal-coating materials to enable nondestructive detection of flaws in coats through inspection under ultraviolet light, according to proposal. Dye-bonding technique prevents outgassing of dyes, making coating materials suitable for use where flaw-free coats must be assured in instrumentation or other applications in which contamination by outgassing must be minimized.

Willis, Paul B.; Cuddihy, Edward F.

1991-01-01

27

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

NASA Astrophysics Data System (ADS)

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.

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

2009-02-01

28

Fluorescence lifetime discrimination of cellular DNA and RNA using various intercalating dyes and flow cytometric analysis  

Microsoft Academic Search

Previously we showed that the fluorescence lifetimes of ethidium and propidium iodide were different when intercalated into cellular double-stranded DNA or RNA. Current studies of four ethidium derivatives showed that the lifetime difference of DNA and RNA bound dyes existed in all four compounds, and that the magnitude of the difference was dependent on the dye structure, the staining concentration,

H. H. Cui; Joseph G. Valdez; John A. Steinkamp; Harry A. Crissman

2001-01-01

29

Fluorescence lifetime discrimination of cellular DNA and RNA using various intercalating dyes and flow cytometric analysis  

NASA Astrophysics Data System (ADS)

Previously we showed that the fluorescence lifetimes of ethidium and propidium iodide were different when intercalated into cellular double-stranded DNA or RNA. Current studies of four ethidium derivatives showed that the lifetime difference of DNA and RNA bound dyes existed in all four compounds, and that the magnitude of the difference was dependent on the dye structure, the staining concentration, and the conditions in which flow cytometry (FCM) analysis of stained cells was performed. The fluorescence lifetime of both DNA and RNA bound fluorochromes was reduced with increasing dye concentration; however the level of decrease varied in different dyes. Further analysis of stained cellular DNA in dye-free solution, which favors only high affinity binding, led to elevated fluorescence lifetime values compared to lifetime values obtained from analysis in the dye solution under equilibrium binding conditions. The changes of fluorescence lifetime value reflect the difference in dye structure and distinct interaction between the dyes and nucleic acids. The staining and analysis variables used in these studies may potentially provide additional information on differences in nucleic acid conformation and function in cell populations.

Cui, H. H.; Valdez, Joseph G.; Steinkamp, John A.; Crissman, Harry A.

2001-05-01

30

Fluorescent indicator dyes for calcium ions  

NASA Technical Reports Server (NTRS)

The present invention discloses a new class of highly fluorescent indicator dyes that are specific for calcium ions. The new fluorescent indicator dyes combine a stilbene-type fluorophore with a tetracarboxylate parent Ca.sup.2+ chelating compound having the octacoordinate pattern of liganding groups characteristic of EGTA and BAPTA. Preferred forms contain extra heterocyclic bridges to reinforce the ethylenic bond of the stilbene and to reduce hydrophobicity. Compared to their widely used predecessor, quin2, the new dyes offer up to thirty-fold brighter fluorescence, major changes in wavelength (not just intensity) upon Ca.sup.2+ binding, slightly lower affinities for Ca.sup.2+, slightly longer wavelengths of excitation, and considerably improved selectivity for Ca.sup.2+ over other divalent cations. These properties, particularly the wavelength sensitivity to Ca.sup.2+, make the dyes useful indicators for many intracellular applications, especially in single cells, adherent cell layers, or bulk tissues. The present invention also discloses an improved method for synthesizing alpha-acyloxyalkyl bromides wherein the bromides so synthesized are free of contaminating bis(1-bromoalkyl)ether. The improved method is exemplified herein in the synthesis of acetoxymethyl bromide, a compound useful in preparing the acetoxymethyl esters disclosed herein as novel Ca.sup.2+ specific fluorescent indicators.

Tsien, Roger Y. (Inventor); Grynkiewicz, Grzegorz (Inventor)

1986-01-01

31

Fluorescence emission spectra of calcofluor stained yeast cell suspensions: heuristic assessment of basis spectra for their linear unmixing.  

PubMed

Fluorescence emission spectra of yeast cell suspensions stained with calcofluor have recently been identified as promising markers of variations in the quality of yeast cell wall. It is shown in this paper how the raw fluorescence spectra of calcofluor can be transformed to reliable spectral signatures of cell wall quality, which are independent of actual dye-to-cell concentrations of examined cell suspensions. Moreover, the presented approach makes it possible to assess basis fluorescence spectra that allows for the spectral unmixing of raw fluorescence spectra in terms of respective fluorescence contributions of calcofluor solvated in the suspension medium and bound to yeast cell walls. PMID:22538834

Plášek, Jaromír; Dostál, Marek; Gášková, Dana

2012-07-01

32

Vital staining from dye-coated microprobes identifies new olfactory interneurons for optical and electrical recording.  

PubMed

A versatile technique for dye application in living tissue is described, which results in labeling of viable cells from which electrophysiological or optical recordings can be obtained. The dye-coated surface of a glass microelectrode tip is used to apply anatomical tracers or calcium sensitive probes with spatial precision. A total of three types of dyes have been applied in this way to find and record from olfactory interneurons in the terrestrial mollusc Limax maximus. Crystals of 1,1'-didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) formed on the tips of glass microelectrodes were placed in the procerebral lobe, the major olfactory processing center of Limax. Somata in buccal and pedal ganglia with processes extending several 100 microm to the procerebral lobe were stained within 4-6 h. Intracellular recordings from DiI stained buccal (B(PC)) and pedal (P(PC)) cells were obtained. Cross correlograms of the oscillatory field potential in the procerebral lobe and spontaneous action potentials in P(PC) or B(PC) show that P(PC) activity is weakly coupled to the oscillation in the procerebral lobe, while B(PC) activity is clearly coupled to the oscillation. Stimulation of the procerebral lobe with nitric oxide activated P(PC) cells but suppressed activity in B(PC) cells. Calcium green-10Kdextran coated electrodes were used to place calcium green in the cell body layer of the procerebral lobe. Bursting and nonbursting procerebral neurons incorporated and transported the calcium green-dextran. Optical recordings of changes in fluorescence signals from several bursting cells recorded simultaneously were used to test alternative mechanisms of bursting cell coupling. Application of biotin 3Kdextran to the midline of the cerebral ganglion revealed a group of cells in each procerebral lobe with processes crossing the midline of the cerebral ganglion. These cells may couple right and left procerebral lobe activity during odor processing. PMID:9128173

Gelperin, A; Flores, J

1997-03-01

33

Perfluorodecalin-soluble fluorescent dyes for the monitoring of circulating nanocapsules with intravital fluorescence microscopy.  

PubMed

Perfluorodecalin (PFD) is an established artificial oxygen carrier due to its physical capability to solve the respiratory gases oxygen and carbon dioxide. PFD-filled poly(n-butyl-cyanoacrylate) (PACA) nanocapsules are already discussed as effective artificial oxygen carriers, and their principal suitability for intravenous administration had been shown. To further elucidate their action in vivo, it is imperative to characterise their preclinical safety and particularly their biodistribution. For these purposes, intravital fluorescence microscopy would display an attractive technique in order to monitor the PACA nanocapsules in vivo, but unfortunately, it is impossible to stain the PACA nanocapsules with a fluorescent dye fulfilling special criteria required for in vivo microscopy. In order to develop such a dye, a long-chained fluorinated thiol was used to modify a BODIPY derivative that is a highly fluorescent organic compound belonging to the difluoro-boraindacene family, as well as to functionalise mesoscopic systems, such as CdSe/ZnS-quantum dots and gold nanoparticles. Furthermore, a functionalisation of porphyrin derivatives was investigated by placing divalent ions in the centre of these systems. Due to the high solubility of all synthesised dyes in PFD, it should be possible to stain PFD-filled particles in general. However, only the functionalised BODIPY derivative was suitable for in vivo monitoring of the PFD-filled PACA nanocapsules. PMID:24963954

Laudien, J; Naglav, D; Gro?-Heitfeld, C; Ferenz, K B; de Groot, H; Mayer, C; Schulz, S; Schnepf, A; Kirsch, M

2014-01-01

34

Evaluation of some fluorescent dyes for water tracing  

Microsoft Academic Search

Eight fluorescent dyes were compared in the laboratory and in field experiments to assess their utility in quantitative tracing work. The properties considered included sensitivity and minimum detectability, the effect of water chemistry on dye fluorescence, photochemical and biological decay rates, adsorption losses on equipment and sediments, toxicity to man and aquatic organisms, and cost. Orange dyes are more useful

P. L. Smart; I. M. S. Laidlaw

1977-01-01

35

Efficacy of Dye-Stained Enteral Formula in Detecting Pulmonary Aspiration  

PubMed Central

Study objective To determine the extent to which a mixture of human gastric juice and enteral formula stained with two concentrations of FD&C Blue No. 1 food dye (0.8 and 1.5 mL/L) is visible in suctioned tracheobronchial secretions following three forced small-volume pulmonary aspirations over a 6-h period in an animal model. Design Experimental 2 × 3 repeated measures. Setting Animal laboratory and an acute care hospital. Participants Ninety New Zealand white rabbits weighing approximately 3 kg each, and 90 acutely ill adults who furnished gastric juice. Interventions A mixture of human gastric juice and enteral formula stained with 0.8 or 1.5 mL of dye per liter was instilled intratracheally over a 30-min period into anesthetized intubated animals at baseline, 2 h, and 4 h. A total of 0.4 mL/kg of the mixture was instilled at each session. Ninety minutes after each instillation, suctioned secretions were examined for visible dye and blood. Measurements and results Dye was visible in 46.3% of the secretions (125 of 270). The concentration of dye had no significant effect on dye visibility. Blood that was present in 114 of 270 of the secretions (42.2%) interfered with dye visibility in all but two secretions. For reasons unknown, even in the absence of blood, dye visibility decreased from 90.2% (55 of 61 secretions) after the first aspiration event to only 61% (25 of 41 secretions) after the third aspiration event. Conclusions Findings from this animal model study do not support the use of the dye method to detect repeated small-volume aspirations. For clinicians who choose to use the dye method in selected situations, it appears that a dye concentration of 0.8 mL/L may be as effective in detecting aspiration as a 1.5 mL/L concentration. PMID:12114370

Metheny, Norma A.; Dahms, Thomas E.; Stewart, Barbara J.; Stone, Kathleen S.; Edwards, Sharon J.; Defer, Julie E.; Clouse, Ray E.

2008-01-01

36

A Method for the Direct Identification of Differentiating Muscle Cells by a Fluorescent Mitochondrial Dye  

PubMed Central

Identification of differentiating muscle cells generally requires fixation, antibodies directed against muscle specific proteins, and lengthy staining processes or, alternatively, transfection of muscle specific reporter genes driving GFP expression. In this study, we examined the possibility of using the robust mitochondrial network seen in maturing muscle cells as a marker of cellular differentiation. The mitochondrial fluorescent tracking dye, MitoTracker, which is a cell-permeable, low toxicity, fluorescent dye, allowed us to distinguish and track living differentiating muscle cells visually by epi-fluorescence microscopy. MitoTracker staining provides a robust and simple detection strategy for living differentiating cells in culture without the need for fixation or biochemical processing. PMID:22174849

Miyake, Tetsuaki; McDermott, John C.; Gramolini, Anthony O.

2011-01-01

37

Van Gieson's picrofuchsin. The staining mechanisms for collagen and cytoplasm, and an examination of the dye diffusion rate model of differential staining.  

PubMed

The staining mechanism of van Gieson's picrofuchsin was studied by use of simple protein model systems and tissue sections, and by spectrophotometry and dialysis experiments. At the endpoint of the staining reaction (equilibrium) cytoplasm is yellow. Dye dilution experiments demonstrated that the highest affinity in the tissue section--picrofuchsin system is between binding sites in cytoplasmic protein and acid fuchsin. Nevertheless sections that were first stained in acid fuchsin (AcF) and then in picrofuchsin ended up with cytoplasm stained yellow. It was concluded that differences in the dye diffusion rates and differences in the permeability of tissue components cannot be invoked to explain the differential staining result. Model experiments with dissolved proteins demonstrated a positive relationship between protein concentration and uptake of picric acid (PA) from picrofuchsin. From this and experiments with additives (sodium dodecylsulphate, urea etc.) and organic solvents, it is proposed that coagulant interchain cross-linking at the high protein concentration of the cytoplasm masks potential dye-binding sites. This affects high affinity dyes with multiple binding sites more than small dyes, and so puts AcF at a disadvantage compared to PA. Staining of non-collagen proteins is mainly by hydrophobic bonding, involving ionic attractions, apolar bonds, and release of water. This mode of binding is relatively strong, decreases swelling and leads to slow dye exchange. Dye binding to collagen is mostly by hydrogen bonds, but in aqueous dye solvent nonpolar residues and charged residues may also participate. This structure remains relatively open during and after dye-binding, and the bound dye ions are therefore easily exchanged for other dye ions. PMID:7683012

Prentø, P

1993-02-01

38

An easy method for cutting and fluorescent staining of thin roots  

PubMed Central

Background and Aims Cutting plant material is essential for observing internal structures and may be difficult for various reasons. Most fixation agents such as aldehydes, as well as embedding resins, do not allow subsequent use of fluorescent staining and make material too soft to make good-quality hand-sections. Moreover, cutting thin roots can be very difficult and time consuming. A new, fast and effective method to provide good-quality sections and fluorescent staining of fresh or fixed root samples, including those of very thin roots (such as Arabidopsis or Noccaea), is described here. Methods To overcome the above-mentioned difficulties the following procedure is proposed: fixation in methanol (when fresh material cannot be used) followed by en bloc staining with toluidine blue, embedding in 6 % agarose, preparation of free-hand sections of embedded material, staining with fluorescent dye, and observation in a microscope under UV light. Key Results Despite eventual slight deformation of primary cell walls (depending on the species and root developmental stage), this method allows effective observation of different structures such as ontogenetic changes of cells along the root axis, e.g. development of xylem elements, deposition of Casparian bands and suberin lamellae in endodermis or exodermis or peri-endodermal thickenings in Noccaea roots. Conclusions This method provides good-quality sections and allows relatively rapid detection of cell-wall modifications. Also important is the possibility of using this method for free-hand cutting of extremely thin roots such as those of Arabidopsis. PMID:22419758

Zelko, Ivan; Lux, Alexander; Sterckeman, Thibault; Martinka, Michal; Kollárová, Karin; Lišková, Desana

2012-01-01

39

Ultrafast fluorescence studies of dye sensitized solar cells.  

PubMed

Time-resolved fluorescence spectra from the RuN719 dye exhibit very short lifetimes (<30 fs) in solutions, on non-injecting substrates and on injecting ones. This reveals <10 fs intramolecular energy redistribution competing with the injection. We conclude that injection proceeds on a sub-10 fs time scale from non-thermalized levels of the dye. PMID:22555159

Bräm, Olivier; Cannizzo, Andrea; Chergui, Majed

2012-06-14

40

NIR fluorescent dyes: versatile vehicles for marker and probe applications  

NASA Astrophysics Data System (ADS)

The use of the NIR spectral region (650-900 nm) is advantageous due to the inherently lower background interference and the high molar absorptivities of NIR chromophores. Near-Infrared (NIR) dyes are increasingly used in the biological and medical field. The binding characteristics of NIR dyes to biomolecules are possibly controlled by several factors, including hydrophobicity, size and charge just to mention a few parameters. Binding characteristics of symmetric carbocyanines and found that the hydrophobic nature of the NIR dye is only partially responsible for the binding strength. Upon binding to biomolecules significant fluorescence enhancement can be observed for symmetrical carbocyanines. This fluorescence amplification facilitates the detection of the NIR dye and enhances its utility as NIR reporter. This manuscript discusses some probe and marker applications of such NIR fluorescent dyes. One application discussed here is the use of NIR dyes as markers. For labeling applications the fluorescence intensity of the NIR fluorescent label can significantly be increased by enclosing several dye molecules in nanoparticles. To decrease self quenching dyes that have relatively large Stokes' shift needs to be used. This is achieved by substituting meso position halogens with amino moiety. This substitution can also serve as a linker to covalently attach the dye molecule to the nanoparticle backbone. We report here on the preparation of NIR fluorescent silica nanoparticles. Silica nanoparticles that are modified with aminoreactive moieties can be used as bright fluorescent labels in bioanalytical applications. A new bioanalytical technique to detect and monitor the catalytic activity of the sulfur assimilating enzyme using NIR dyes is reported as well. In this spectroscopic bioanalytical assay a family of Fischer based n-butyl sulfonate substituted dyes that exhibit distinct variation in absorbance and fluorescence properties and strong binding to serum albumin as its sulfonic acid moiety is modified to less water soluble moiety was identified. In polar solvents, these water soluble compounds are strongly fluorescent, however form the less soluble aggregated species with virtual loss of fluorescence when the sulfonate groups are cleaved by enzymatic activity to form the corresponding straight chain alkyl aldehyde derivatives. To achieve this conversion in vitro photo-reduced riboflavin mononucleotide (FMN) with a glucose/ glucose-oxygenase oxygen scavenging system was utilized. The reduced FMN serves as a key substrate in the enzymatic desulfonation. Once the FMNH2 was produced the desulfonation reaction was characterized by using Laser Induced Fluorescence Capillary Zone Electropheresis (LIF-CZE). This method can be utilized as an assay to detect the enzyme activity in vitro with the possibilities of in vivo applications.

Patonay, Gabor; Chapman, Gala; Beckford, Garfield; Henary, Maged

2013-02-01

41

WASTES FROM MANUFACTURE OF DYES AND PIGMENTS. VOLUME 3. STILBENE DYES AND FLUORESCENT BRIGHTENING AGENTS  

EPA Science Inventory

A preliminary study of the manufacture of Stilbene dyes and fluorescent brightening agents was conducted to determine if process waste streams might contain hazardous material. The study first identifies the dyes and pigments that belong to this segment of the industry, the amoun...

42

Optical imaging of kidney cancer with novel near-infrared heptamethine carbocyanine fluorescent dyes  

PubMed Central

PURPOSE To assess the application of near-infrared (NIR) heptamethine carbocyanine dyes, IR-783 and the synthetic analog MHI-148, as optical imaging agents for rapid detection of human kidney cancer. MATERIALS AND METHODS The uptake, retention and subcellular localization of these organic dyes were investigated in cultured kidney cancer cells. Tumor specificity of dye uptake and retention was evaluated by whole-body imaging of mice bearing human kidney cancer xenografts or freshly harvested clinical kidney cancer specimens. In addition, dye accumulation at the tissue and cellular levels was confirmed by ex vivo studies with results confirmed by fluorescence imaging of the frozen tissue sections. Peripheral blood spiked with kidney cancer cells was stained to simulate the detection of circulating tumor cells. RESULTS Preferential uptake and retention of carbocyanine NIR dyes was observed in cultured human kidney cancer cells, human kidney cancer cell-spiked whole blood, human kidney cancer xenografts and freshly harvested human kidney cancer tissues compared to normal kidney epithelial cells or normal host organs. CONCLUSIONS We described a new class of NIR heptamethine carbocyanine dyes showing potential for detecting kidney cancer cells in circulating blood and kidney cancer cells in clinical specimens. NIR carbocyanine dyes can be further developed as dual modality agents for deep-tissue imaging of localized and disseminated kidney cancer in patients. PMID:23000848

Wang, Ruoxiang; Chu, Chia-Yi; Hu, Peizhen; Master, Viraj; Osunkoya, Adeboye O.; Kim, Hyung L; Zhau, Haiyen E.; Chung, Leland W. K.

2014-01-01

43

Development of a Vital Fluorescent Staining Method for Monitoring Bacterial Transport in Subsurface Environments  

Microsoft Academic Search

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies

MARK E. FULLER; SHERYL H. STREGER; RANDI K. ROTHMEL; BRIAN J. MAILLOUX; JAMES A. HALL; TULLIS C. ONSTOTT; JAMES K. FREDRICKSON; DAVID L. BALKWILL; MARY F. DEFLAUN

2000-01-01

44

Near-infrared fluorescence imaging using organic dye nanoparticles.  

PubMed

Near-infrared (NIR) fluorescence imaging in the 700-1000 nm wavelength range has been very attractive for early detection of cancers. Conventional NIR dyes often suffer from limitation of low brightness due to self-quenching, insufficient photo- and bioenvironmental stability, and small Stokes shift. Herein, we present a strategy of using small-molecule organic dye nanoparticles (ONPs) to encapsulate NIR dyes to enable efficient fluorescence resonance energy transfer to obtain NIR probes with remarkably enhanced performance for in vitro and in vivo imaging. In our design, host ONPs are used as not only carriers to trap and stabilize NIR dyes, but also light-harvesting agent to transfer energy to NIR dyes to enhance their brightness. In comparison with pure NIR dyes, our organic dye nanoparticles possess almost 50-fold increased brightness, large Stokes shifts (?250 nm) and dramatically enhanced photostability. With surface modification, these NIR-emissive organic nanoparticles have water-dispersity and size- and fluorescence- stability over pH values from 2 to 10 for almost 60 days. With these superior advantages, these NIR-emissive organic nanoparticles can be used for highly efficient folic-acid aided specific targeting in vivo and ex vivo cellular imaging. Finally, during in vivo imaging, the nanoparticles show negligible toxicity. Overall, the results clearly display a potential application of using the NIR-emissive organic nanoparticles for in vitro and in vivo imaging. PMID:24461324

Yu, Jia; Zhang, Xiujuan; Hao, Xiaojun; Zhang, Xiaohong; Zhou, Mengjiao; Lee, Chun-Sing; Chen, Xianfeng

2014-03-01

45

Visualizing Endocytotic Pathways at Transmission Electron Microscopy via Diaminobenzidine Photo-Oxidation by a Fluorescent Cell-Membrane Dye  

PubMed Central

The endocytotic pathway involves a complex, dynamic and interacting system of intracellular compartments. PKH26 is a fluorescent dye specific for long-lasting cell membrane labelling which has been successfully used for investigating cell internalization processes, at either flow cytometry or fluorescence microscopy. In the present work, diaminobenzidine photo-oxidation was tested as a procedure to detect PKH26 dye at transmission electron microscopy. Our results demonstrated that DAB photo-oxidation is a suitable technique to specifically visualise this fluorescent dye at the ultrastructural level: the distribution of the granular dark reaction product perfectly matches the pattern of the fluorescence staining, and the electron density of the fine precipitates makes the signal evident and precisely detectable on the different subcellular compartments involved in the plasma membrane internalization routes. PMID:25578976

Grecchi, S.; Malatesta, M.

2014-01-01

46

Dimeric fluorescent energy transfer dyes comprising asymmetric cyanine azole-indolenine chromophores  

DOEpatents

Novel fluorescent heterodimeric DNA-staining energy transfer dyes are provided combining asymmetric cyanine azole-indolenine dyes, which provide for strong DNA affinity, large Stokes shifts and emission in the red region of the spectrum. The dyes find particular application in gel electrophoresis and for labels which may be bound to a variety of compositions in a variety of contexts. Kits and individual compounds are provided, where the kits find use for simultaneous detection of a variety of moieties, particularly using a single narrow wavelength irradiation source. The individual compounds are characterized by high donor quenching and high affinity to dsDNA as a result of optimizing the length of the linking group separating the two chromophores.

Glazer, Alexander N. (Orinda, CA); Benson, Scott C. (Oakland, CA)

1998-01-01

47

Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining  

PubMed Central

Summary The article describes the immobilization of different probe oligonucleotides (4, 7, 10) carrying each a racemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol (1a) at the 5’-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The bilayer separates two compartments (cis/trans channel) of an optical transparent microfluidic sample carrier with perfusion capabilities. Injection of unlabeled target DNA sequences (6, 8, or 9), differing in sequence and length, leads in the case of complementarity to the formation of stable DNA duplexes at the bilayer surface. This could be verified by Sybr Green I double strand staining, followed by incubation periods and thorough perfusions, and was visualized by single molecule fluorescence spectroscopy and microscopy. The different bilayer-immobilized complexes consisting of various DNA duplexes and the fluorescent dye were studied with respect to the kinetics of their formation as well as to their stability against perfusion. PMID:25298798

Werz, Emma

2014-01-01

48

Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining.  

PubMed

The article describes the immobilization of different probe oligonucleotides (4, 7, 10) carrying each a racemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol (1a) at the 5'-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The bilayer separates two compartments (cis/trans channel) of an optical transparent microfluidic sample carrier with perfusion capabilities. Injection of unlabeled target DNA sequences (6, 8, or 9), differing in sequence and length, leads in the case of complementarity to the formation of stable DNA duplexes at the bilayer surface. This could be verified by Sybr Green I double strand staining, followed by incubation periods and thorough perfusions, and was visualized by single molecule fluorescence spectroscopy and microscopy. The different bilayer-immobilized complexes consisting of various DNA duplexes and the fluorescent dye were studied with respect to the kinetics of their formation as well as to their stability against perfusion. PMID:25298798

Werz, Emma; Rosemeyer, Helmut

2014-01-01

49

Ultrafast dynamics of a new class of highly fluorescent boron difluoride dyes.  

PubMed

The first ultrafast study of the dimeric fluorescent BF2 dye BOPHY is presented. When compared to a structurally related BODIPY dye, similar photophysical dynamics are observed, including an intermediate kinetic component present in both dye types. PMID:25500795

Wang, L; Tamgho, I-S; Crandall, L A; Rack, J J; Ziegler, C J

2015-01-28

50

Nonlinear Emission of Quinolizinium-Based Dyes with Application in Fluorescence Lifetime Imaging.  

PubMed

Charged molecules based on the quinolizinum cation have potential applications as labels in fluorescence imaging in biological media under nonlinear excitation. A systematic study of the linear and nonlinear photophysics of derivatives of the quinolizinum cation substituted by either dimethylaniline or methoxyphenyl electron donors is performed. The effects of donor strength, conjugation length, and symmetry in the two-photon emission efficiency are analyzed in detail. The best performing nonlinear fluorophore, with two-photon absorption cross sections of 1140 GM and an emission quantum yield of 0.22, is characterized by a symmetric D-?-A(+)-?-D architecture based on the methoxyphenyl substituent. Application of this molecule as a fluorescent marker in optical microscopy of living cells revealed that, under favorable conditions, the fluorophore can be localized in the cytoplasmatic compartment of the cell, staining vesicular shape organelles. At higher dye concentrations and longer staining times, the fluorophore can also penetrate into the nucleus. The nonlinearly excited fluorescence lifetime imaging shows that the fluorophore lifetime is sensitive to its location in the different cell compartments. Using fluorescence lifetime microscopy, a multicolor map of the cell is drafted with a single dye. PMID:25135761

Marcelo, Gema; Pinto, Sandra; Cañeque, Tatiana; Mariz, Inês F A; Cuadro, Ana M; Vaquero, Juan J; Martinho, José M G; Maçôas, Ermelinda M S

2014-09-01

51

Development of Highly Fluorescent Materials Based on Thiophenylimidazole Dyes  

NASA Technical Reports Server (NTRS)

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

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

2000-01-01

52

Electrowetting actuation of a dye-doped fluorescent droplet  

NASA Astrophysics Data System (ADS)

We report tunable color output from a fluorescent dye-doped droplet actuated by electrowetting. The system design, based on a planar electrowetting set-up, is compact and straightforward, with minimal voltage requirements for effective actuation. Fluorescent droplets are sourced from a 1 mM solution of rhodamine 6G in distilled water. Initial contact angle for a dye-doped droplet is 72.1°. At a maximum applied voltage of 20 V, the contact angle decreases to 56.5°. Emission spectra are collected as the droplet fluoresces under UV illumination. Over an electrowetting voltage range of 0 to 20 V, the peak fluorescence wavelength shifts from 568 to 546 nm.

Guerrero, Raphael A.; Mero, Rea Divina C.

2014-11-01

53

Disposable nitrate-selective optical sensor based on fluorescent dye  

Technology Transfer Automated Retrieval System (TEKTRAN)

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

54

Approximate analytic solutions for the optical pumping of fluorescent dyes  

NASA Technical Reports Server (NTRS)

A general technique for solving a system of rate equations describing the interaction of an electromagnetic field and a molecular system is presented. The method is used to obtain approximate time-dependent solutions for the upper-level population of fluorescent dyes in the presence of a pump field.

Lawandy, N. M.

1978-01-01

55

High-sensitivity capillary electrophoresis of double-stranded DNA fragments using monomeric and dimeric fluorescent intercalating dyes  

SciTech Connect

Fluorescence-detected capillary electrophoresis separations of [phi]X174/HaeIII DNA restriction fragments have been performed using monomeric and dimeric intercalating dyes. Replaceable hydroxyethyl cellulose solutions were used as the separation medium. Confocal fluorescence detection was performed following 488-nm laser excitation. The limits of DNA detection for on-column staining with monomeric dyes (ethidium bromide, two propidium dye derivatives, oxazole yellow, thiazole orange, and a polycationic thiazole orange derivative) were determined. The thiazole orange dyes provide the most sensitive detection with limiting sensitivities of 2-4 amol of DNA base pairs per band, and detection of the 603-bp fragment was successful, injecting from [phi]X174/HaeIII samples containing only 1-2 fg of this fragment per microliter. Separations of preformed DNA-dimeric dye complexes were also performed. The breadth of the bands observed in separations of preformed DNA-dimeric dye complexes is due to the presence of DNA fragments with different numbers of bound dye molecules that can be resolved as closely spaced subbands in many of our separations. The quality of these DNA-dye complex separations can be dramatically improved by performing the electrophoresis with 9-aminoacridine (9AA) in the column and running buffers. 43 refs., 10 figs., 1 tab.

Zhu, H.; Clark, S.M.; Benson, S.C.; Rye, H.S.; Glazer, A.N.; Mathies, R.A. (Univ. of California, Berkeley, CA (United States))

1994-07-01

56

A berberine-aniline blue fluorescent staining procedure for suberin, lignin, and callose in plant tissue  

Microsoft Academic Search

Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining

Mark C. Brundrett; Daryl E. Enstone; Carol A. Peterson

1988-01-01

57

Inhibition of beta-amyloid aggregation by fluorescent dye labels  

NASA Astrophysics Data System (ADS)

The fluorescence decay of beta-amyloid's (A?) intrinsic fluorophore tyrosine has been used for sensing the oligomer formation of dye-labelled A? monomers and the results compared with previously studied oligomerization of the non-labelled A? peptides. It has been demonstrated that two different sized, covalently bound probes 7-diethylaminocoumarin-3-carbonyl and Hilyte Fluor 488 (HLF), alter the rate and character of oligomerization to different extents. The ability of HLF to inhibit formation of highly ordered structures containing beta-sheets was also shown. The implications of our findings for using fluorescence methods in amyloidosis research are discussed and the advantages of this auto-fluorescence approach highlighted.

Amaro, Mariana; Wellbrock, Thorben; Birch, David J. S.; Rolinski, Olaf J.

2014-02-01

58

Rapid enumeration of respiratory active mycobacteria with fluorescent double staining.  

PubMed

A new method for rapid enumeration of physiologically active mycobacteria was developed by acid-fast bacilli staining (Auramine O) following formazan reduction. Results can be obtained within 90 min by the optimized procedure, while more than one week is required for the widely used culture-dependent approach. PMID:20600362

Ichijo, Tomoaki; Izumi, Yoko; Yamaguchi, Nobuyasu; Nasu, Masao

2010-09-01

59

Fluorescence of Alexa Fluor Dye Tracks Protein Folding  

PubMed Central

Fluorescence spectroscopy is an important tool for the characterization of protein folding. Often, a protein is labeled with appropriate fluorescent donor and acceptor probes and folding-induced changes in Förster Resonance Energy Transfer (FRET) are monitored. However, conformational changes of the protein potentially affect fluorescence properties of both probes, thereby profoundly complicating interpretation of FRET data. In this study, we assess the effects protein folding has on fluorescence properties of Alexa Fluor 488 (A488), which is commonly used as FRET donor. Here, A488 is covalently attached to Cys69 of apoflavodoxin from Azotobacter vinelandii. Although coupling of A488 slightly destabilizes apoflavodoxin, the three-state folding of this protein, which involves a molten globule intermediate, is unaffected. Upon folding of apoflavodoxin, fluorescence emission intensity of A488 changes significantly. To illuminate the molecular sources of this alteration, we applied steady state and time-resolved fluorescence techniques. The results obtained show that tryptophans cause folding-induced changes in quenching of Alexa dye. Compared to unfolded protein, static quenching of A488 is increased in the molten globule. Upon populating the native state both static and dynamic quenching of A488 decrease considerably. We show that fluorescence quenching of Alexa Fluor dyes is a sensitive reporter of conformational changes during protein folding. PMID:23056480

Lindhoud, Simon; Westphal, Adrie H.; Visser, Antonie J. W. G.; Borst, Jan Willem; van Mierlo, Carlo P. M.

2012-01-01

60

IR-780 Dye for Near-Infrared Fluorescence Imaging in Prostate Cancer.  

PubMed

Background The aim of this study was to investigate near-infrared fluorescence (NIRF) imaging as a novel imaging modality that allows for early detection of cancer and real-time monitoring to acquire related information. IR-780 iodide, a lipophilic dye, accumulates selectively in breast cancer cells and drug-resistant human lung cancer cells, with a peak emission at 780 nm that can be easily detected by the NIRF imaging system. The application of IR-780 for prostate cancer imaging was thoroughly investigated to further expand its clinical value. Material and Methods The impact of IR-780 on the survival of prostate cancer cells PC-3 and LNCaP as well as normal prostate epithelial cells RWPE-1 was determined. Duration of IR-780 dye staining was optimized in PC-3 cells. The involvement of specific OATP1B3 inhibitor in the selective accumulation of IR-780 was investigated. IR-780 for prostate cancer imaging was carried out in athymic nude mouse models and, acute toxicity of IR-780 was evaluated. Results IR-780 incubation resulted in a dose-dependent inhibition to cell proliferation. Mean fluorescence intensity of prostate cancer cells peaked at 20-min IR-780 incubation. Specific uptake of IR-780 dye in prostate cancer cells was mainly through the function of OATP1B3. We also demonstrated that NIRF dye effectively identified the subcutaneous prostate cancer xenografts, subsequently confirmed by histological examination. There was no significant impact on the physical activity, weight, and tissue histology of BABL/C mice with 10-fold imaging dose of 1-month IR-780 dye administration. Conclusions NIRF imaging using IR-780 dye is a feasible and practicable method for prostate cancer detection, with potential tumor-killing ability, although more investigations are needed before clinical translation. PMID:25686161

Yi, Xiaomin; Yan, Fei; Wang, Fuli; Qin, Weijun; Wu, Guojun; Yang, Xiaojian; Shao, Chen; Chung, Leland W K; Yuan, Jianlin

2015-01-01

61

Near-infrared fluorescence imaging of prostate cancer using heptamethine carbocyanine dyes  

PubMed Central

Near-infrared fluorescence (NIRF) imaging is an attractive novel modality for the detection of cancer. A previous study defined two organic polymethine cyanine dyes as ideal NIRF probes, IR-783 and its derivative MHI-148, which have excellent optical characteristics, superior biocompatibility and cancer targeting abilities. To investigate the feasibility of NIRF dye-mediated prostate cancer imaging, dye uptake and subcellular co-localization were investigated in PC-3, DU-145 and LNCaP human prostate cancer cells and RWPE-1 normal prostate epithelial cells. Different organic anion transporting peptide (OATP) inhibitors were utilized to explore the potential role of the OATP subtype, including the nonspecific OATP inhibitor bromosulfophthalein, the OATP1 inhibitor 17?-estradiol, the selective OATP1B1 inhibitor rifampicin and the selective OATP1B3 inhibitor cholecystokinin octapeptide. NIRF dyes were also used for the simulated detection of circulating tumor cells and the rapid detection of prostate cancer in human prostate cancer tissues and prostate cancer xenografts in mouse models. The results revealed that the cancer-specific uptake of these organic dyes in prostate cancer cells occurred primarily via OATP1B3. A strong NIRF signal was detected in prostate cancer tissues, but not in normal tissues that were stained with IR-783. Prostate cancer cells were recognized with particular NIR fluorescence in isolated mononuclear cell mixtures. The results of the present study demonstrated that NIRF dye-mediated imaging is a feasible and practicable method for prostate cancer detection, although further investigative studies are required before clinical translation. PMID:25354708

YUAN, JIANLIN; YI, XIAOMIN; YAN, FEI; WANG, FULI; QIN, WEIJUN; WU, GUOJUN; YANG, XIAOJIAN; SHAO, CHEN; CHUNG, LELAND W.K.

2015-01-01

62

Near?infrared fluorescence imaging of prostate cancer using heptamethine carbocyanine dyes.  

PubMed

Near?infrared fluorescence (NIRF) imaging is an attractive novel modality for the detection of cancer. A previous study defined two organic polymethine cyanine dyes as ideal NIRF probes, IR?783 and its derivative MHI?148, which have excellent optical characteristics, superior biocompatibility and cancer targeting abilities. To investigate the feasibility of NIRF dye?mediated prostate cancer imaging, dye uptake and subcellular co?localization were investigated in PC?3, DU?145 and LNCaP human prostate cancer cells and RWPE?1 normal prostate epithelial cells. Different organic anion transporting peptide (OATP) inhibitors were utilized to explore the potential role of the OATP subtype, including the nonspecific OATP inhibitor bromosulfophthalein, the OATP1 inhibitor 17??estradiol, the selective OATP1B1 inhibitor rifampicin and the selective OATP1B3 inhibitor cholecystokinin octapeptide. NIRF dyes were also used for the simulated detection of circulating tumor cells and the rapid detection of prostate cancer in human prostate cancer tissues and prostate cancer xenografts in mouse models. The results revealed that the cancer?specific uptake of these organic dyes in prostate cancer cells occurred primarily via OATP1B3. A strong NIRF signal was detected in prostate cancer tissues, but not in normal tissues that were stained with IR?783. Prostate cancer cells were recognized with particular NIR fluorescence in isolated mononuclear cell mixtures. The results of the present study demonstrated that NIRF dye?mediated imaging is a feasible and practicable method for prostate cancer detection, although further investigative studies are required before clinical translation. PMID:25354708

Yuan, Jianlin; Yi, Xiaomin; Yan, Fei; Wang, Fuli; Qin, Weijun; Wu, Guojun; Yang, Xiaojian; Shao, Chen; Chung, Leland W K

2015-02-01

63

Fluorescent labeling of dendritic spines in cell cultures with the carbocyanine dye "DiI".  

PubMed

Analyzing cell morphology is a key component to understand neuronal function. Several staining techniques have been developed to facilitate the morphological analysis of neurons, including the use of fluorescent markers, such as DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). DiI is a carbocyanine membrane dye that exhibits enhanced fluorescence upon insertion of its lipophilic hydrocarbon chains into the lipid membrane of cells. The high photostability and prominent fluorescence of the dye serves as an effective means of illuminating cellular architecture in individual neurons, including detailed dendritic arborizations and spines in cell culture and tissue sections. Here, we specifically optimized a simple and reliable method to fluorescently label and visualize dissociated hippocampal neurons using DiI and high-resolution confocal microscopic imaging. With high efficacy, this method accurately labels neuronal and synaptic morphology to permit quantitative analysis of dendritic spines. Accurate imaging techniques of these fine neuronal specializations are vital to the study of their morphology and can help delineate structure-function relationships in the central nervous system. PMID:24847216

Cheng, Connie; Trzcinski, Olivia; Doering, Laurie C

2014-01-01

64

In situ preparation of highly fluorescent dyes upon photoirradiation.  

PubMed

Photoswitchable or photoactivatable fluorescent dyes are potentially applicable to ultrahigh density optical memory media as well as super-resolution fluorescence imaging when the dyes are highly fluorescent and have large absorption coefficients. Here, we report on highly fluorescent photochromic dyes, which are initially nonluminous in solution under irradiation with visible light but activated to emit green or red fluorescence upon irradiation with ultraviolet (UV) light. The dyes 5a-9a are sulfone derivatives of 1,2-bis(2-ethyl-6-phenyl(or thienyl)-1-benzothiophen-3-yl)perfluorocyclopentene. It was found that substitution of phenyl or thiophene rings at 6 and 6' positions of the benzothiophene-1,1-dioxide groups is effective to increase the fluorescence quantum yields of the closed-ring isomers over 0.7 and absorption coefficients over 4 × 10(4) M(-1) cm(-1). The phenyl-substituted derivatives 5a-7a undergo photocyclization reactions to produce yellow closed-ring isomers 5b-7b, which emit brilliant green fluorescence at around 550 nm (?(F) = 0.87-0.88) under irradiation with 488 nm light. Any absorption intensity change of the closed-ring isomers was not observed even after 100 h storage in the dark at 80 °C. The closed-ring isomers slowly returned to the initial open-ring isomers upon irradiation with visible (? > 480 nm) light. The ring-opening quantum yields (?(C?O)) were measured to be (1.6-4.0) × 10(-4). When the phenyl substituents are replaced with thiophene rings, such as compounds 8a and 9a, the absorption bands of the closed-ring isomers shift to longer than 500 nm. The closed-ring isomers exhibit brilliant red fluorescences at around 620 nm (?(F) = 0.61-0.78) under irradiation with 532 nm light. The ring-opening reactions are very slow (?(C?O) < 1 × 10(-5)). The fluorescence lifetimes of these sulfone derivatives were measured to be around 2-3 ns, which is much longer than the value of the closed-ring isomer of 1,2-bis(2-methyl-1-benzothiophen-3-yl)perfluorocyclopentene (?(F) = 4 and 22 ps). The closed-ring isomer 8b in 1,4-dioxane exhibits excellent fatigue resistant property under irradiation with visible light (? > 440 nm) superior to the stability of Rhodamine 101 in ethanol. PMID:21819048

Uno, Kakishi; Niikura, Hiroyuki; Morimoto, Masakazu; Ishibashi, Yukihide; Miyasaka, Hiroshi; Irie, Masahiro

2011-08-31

65

Assessment of gamete quality for the eastern oyster (Crassostrea virginica) by use of fluorescent dyes  

USGS Publications Warehouse

Evaluation of sperm motility is the single most widely used parameter to determine semen quality in mammals and aquatic species. While a good indicator for fresh sperm viability, post-thaw motility is not always effective at predicting fertilizing ability. Techniques using fluorescent dyes can assess functionality of mammalian sperm, but have not been widely applied in aquatic organisms. The eastern oyster Crassostrea virginica is an important mollusk in the United States, and cryopreservation protocols have been developed to preserve sperm and larvae to assist research and hatchery production. In this study, protocols were developed to assess sperm cell membrane integrity and mitochondrial function by flow cytometry and to assess viability of eggs by fluorescence microscopy. The fluorescent dyes SYBR 14 and propidium iodide (PI) (to assess membrane integrity) and rhodamine 123 (R123) (to assess mitochondrial membrane potential) were used to evaluate the quality of thawed oyster sperm previously cryopreserved with different cryoprotectant and thawing treatments. Membrane integrity results were correlated with motility of thawed sperm and mitochondrial membrane potential with fertilizing ability. Fluorescein diacetate (FDA) was used to assess cytotoxicity of cryoprotectant solutions and post-thaw damage to oyster eggs. The results indicated that membrane integrity (P = 0.004) and thawing treatments (P = 0.04), and mitochondrial membrane potential (P = 0.0015) were correlated with motility. Fertilizing ability was correlated with cryoprotectant treatments (P = 0.0258) and with mitochondrial membrane potential (P = 0.001). The dye FDA was useful in indicating structural integrity of fresh and thawed eggs. Exposure of eggs, without freezing, to dimethyl sulfoxide yielded higher percentages of stained eggs and fertilization rate than did exposure to propylene glycol (P = 0.002). Thawed eggs were not stained with FDA (<1%) and larvae were not produced. ?? 2006 Elsevier Inc. All rights reserved.

Paniagua-Chavez, C. G.; Jenkins, J.; Segovia, M.; Tiersch, T.R.

2006-01-01

66

Precise fluorescence measurement for determination of photophysical properties of dyes  

NASA Astrophysics Data System (ADS)

The authors present a double frequency modulation technique to determine the fluorescence power with an accuracy of at least 10 -5. Quantitative analysis of the modulation transfer from the excitation to the fluorescence power is used to derive the triplet state lifetime ?DS and the rate constant for intersystem crossing kED of fluorescent dyes. For that purpose the correlation between the population of transient states and the decrease of fluorescence power is measured. The experimental results of rhodamine 6G are presented. The influence of molecular oxygen is also examined. The photophysical parameters can be determined with high accuracy even if absorption of excited states, photoproducts or other molecules is present. This sensitivity is sufficient to determine the presence of a few hundred molecules in excited states.

Gregor, I.; Heupel, Ma.; Thiel, E.

2001-10-01

67

Fluorescent performance of electrospun polyimide web mixed with hemicyanine dye  

Microsoft Academic Search

Polyamic acid (PAA) solution in N, N-dimethylacetamide was mixed with one hemicyanine dye, DHEASPI-C1, to prepare the fluorescent polyimide (PI) web by electrospinning process (ESP). Firstly, dark orange-red PAA\\/DHEASPI-C1 web consisted of nanofibers about 10–100 nm, could be achieved using ESP at room temperature. Then thermal imidization for 4 h in vacuum oven (200 °C, 133 Pa) led to the cycloimidization of PAA into

Chuanxiang Qin; Jianjun Wang; Si Cheng; Xiaomei Wang; Lixing Dai; Guoqiang Chen

2009-01-01

68

Functionalization of Cloisite 30B with fluorescent dyes  

Microsoft Academic Search

An organo-montmorillonite (Cloisite® 30B) was modified with fluorescent dyes to obtain photo-functional inorganic–organic complexes for real-time monitoring of polymer-clay nanocomposite processing. The photo-functionalization with 9-anthracenemethanol (anth) was tested using adsorption from solution, as well as dry and melt compounding. Rhodamine 6G Perchlorate (RhP) and Nile Blue A Perchlorate (NBAP) were adsorbed by cation exchange. XRD, TGA and spectrofluorimetry showed that

O. Raccurt; J.-Y. Charmeau; J. Duchet-Rumeau

2010-01-01

69

Fluorescent Staining of Tea Pathogenic Fungi in Tea Leaves Using Fluorescein-labeled Lectin  

NASA Astrophysics Data System (ADS)

Fluorochrome-labeled lectin, fluorescein conjugated wheat germ agglutinin (F-WGA) was applied to stain tea pathogenic fungi in tea leaf tissue. Infected leaves were fixed and decolorized with a mixture of ethanol and acetic acid, and cleared with 10% KOH for whole mount before staining with F-WGA. Hyphae of Pestalotiopsis longiseta, Pseudocercospora ocellata, Botrytis cinerea and Colletotrichum theae-sinensis fluoresced brightly in whole mount and sectioned samples of infected leaf tissue. In browned tissue, hyphae did not fluoresce frequently in whole mount sample. Autofluorescence of leaf tissue was strong in browned tissue of sections, it was removed by 10% KOH treatment before staining. Penetration hyphae of C. theae-sinensis in cell wall of trichome and hyphae in basal part of trichome did not fluoresced frequently. In whole mount samples of tea leaf infected with Exobasidium vexans and E. reticulatum, hymenia appeared on leaf surface fluoresced, but hyphae in leaf tissue did not fluoresce. In sectioned samples, hyphae fluoresced brightly when sections were treated with 10% KOH before staining.

Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

70

Deterministically Polarized Fluorescence from Single Dye Molecules Aligned in Liquid Crystal Host  

SciTech Connect

We demonstrated for the first time to our konwledge deterministically polarized fluorescence from single dye molecules. Planar aligned nematic liquid crystal hosts provide deterministic alignment of single dye molecules in a preferred direction.

Lukishova, S.G.; Schmid, A.W.; Knox, R.; Freivald, P.; Boyd, R. W.; Stroud, Jr., C. R.; Marshall, K.L.

2005-09-30

71

Ethidium bromide: a fast fluorescent staining procedure for the detection of symbiotic partnership of flagellates and prokaryotes.  

PubMed

The hindgut of 'lower' termites harbors a dense population of flagellates and bacteria. The flagellates possess ecto- and endosymbiotic prokaryotes. Most of them are hardly visible in the phase contrast microscope. Staining with the DNA-intercalating agent ethidium bromide visualizes the nuclei of the flagellates as well as the ecto- and endosymbiotic bacteria as red objects. Furthermore, it is possible to distinguish between endosymbiotic methanogens and other bacteria. Following UV excitation, the blue-green autofluorescence of the methanogenic bacteria eclipses the red fluorescence light of the intercalated ethidium bromide. The dye facilitates the observation of symbiotic bacteria and helps identify the number, shape, localization, and dividing status of the nuclei. Thus, it is a powerful tool for the examination of microorganisms in complex habitats, which are rich in strongly autofluorescent material, like wood. PMID:10192044

Fröhlich, J; König, H

1999-03-01

72

Stain Reagent Reversible Stain Kits  

E-print Network

GelCode ® Blue Stain Reagent GelCode ® SilverSNAP TM Stain GelCode ® E-Zinc TM Reversible Stain ease of use and relatively good sensitivity. Although alternative methods, such as silver staining,1 as a colloidal suspension. The colloidal particles do not penetrate the gel, but the dye molecules are extracted

Lebendiker, Mario

73

A double filtering method for measuring the translational velocity of fluorescently stained cells  

NASA Astrophysics Data System (ADS)

The authors propose a double filtering method to measure translational velocity for tracking fluorescently stained cells. This method employs two diffraction gratings installed in the infinity space through which the parallel pencil beam of the fluorescence passes. With this method, the change in light intensity whose period is proportional to the translational velocity of the sample can be obtained at the imaging surface. By using a sample that has a random distribution of fluorescence intensity, the authors verified that translational velocity measurements could be achieved using the proposed method.

Yasokawa, Toshiki; Ishimaru, Ichirou; Kuriyama, Shigeki; Masaki, Tsutomu; Takegawa, Kaoru; Tanaka, Naotaka

2007-09-01

74

Vital staining from dye-coated microprobes identifies new olfactory interneurons for optical and electrical recording  

Microsoft Academic Search

A versatile technique for dye application in living tissue is described, which results in labeling of viable cells from which electrophysiological or optical recordings can be obtained. The dye-coated surface of a glass microelectrode tip is used to apply anatomical tracers or calcium sensitive probes with spatial precision. A total of three types of dyes have been applied in this

A Gelperin; J Flores

1997-01-01

75

A New Fluorescence Staining Assay for Visualizing Living Microorganisms in Soil  

PubMed Central

5- (and 6-)Sulfofluorescein diacetate (SFDA), which is converted to a fluorescent product by intracellular esterase activity, was used to stain living microorganisms, including bacteria, Saccharomyces cerevisiae, and fungi, in soil. SFDA (1 mM) dissolved in ethyl alcohol was added to an intact soil sample, and the preparation was examined with an epifluorescence microscope. Bright single cells and colonies of live bacteria were observed without interference from the autofluorescence of soil minerals and detritus. Cultured Escherichia coli was killed through heat treatment; thus, SFDA was concluded to stain only living cells. Microbial colonies obtained from natural soils and various cultured strains were tested. It was found that 151 of 154 colonies from natural soils were stained and that hyphae and spores from 1 of 28 cultured microbial strains were not stained. The SFDA method was successfully used to visualize and count bacteria in soil samples from Mount Shiga in Japan. PMID:16535127

Tsuji, T.; Kawasaki, Y.; Takeshima, S.; Sekiya, T.; Tanaka, S.

1995-01-01

76

Fluorescent liposomes for intravital staining of Kupffer cells to aid in vivo microscopy in rats.  

PubMed

The potential of newly formulated fluorescent-labeled liposomes for the intravital staining of Kupffer cells was evaluated in rats. Fluorescently labeled phosphatidylcholine (PC) was incorporated into liposomes consisting of PC and phosphatidylserine. After intravenous injection, Kupffer cells in the rat liver were intravitally stained and were clearly delineated under the fluorescence image of both confocal laser scanning microscopy and in vivo microscopy. Specificity of the staining was confirmed by immunohistochemistry using the anti-rat macrophage antibody Ki-M2R, which suggested that the liposomes were selectively entrapped by the hepatic reticuloendothelial system. A time-course study revealed that the suitable observation window was between 16 and 24 h after the injection. Phagocytic activity of Kupffer cells after the administration of liposomes was examined by measuring the amount of hepatic uptake of intravenously administered fluorescent microspheres; no detrimental influence of the liposomes on the phagocytic activity was observed. Additionally, no histopathologic changes were found in the livers from liposome-treated rats. Therefore, the fluorescent-labeled liposomes appear to be a useful research tool for labeling Kupffer cells for in vivo microscopic observation of the liver. PMID:17805433

Watanabe, R; Munemasa, T; Matsumura, M; Fujimaki, M

2007-06-01

77

Engineering metal-nanoantennae/dye complexes for maximum fluorescence enhancement.  

PubMed

We theoretically investigate the fluorescence enhancement of a molecule placed in a variable (4 - 20 nm) gap of a plasmonic dimer, with different dye molecules as well as different nanoparticle geometries, using a fully vectorial three-dimensional finite-difference time-domain (3D FDTD) method. This work extends previous studies on molecular fluorescence in the vicinity of metal interfaces and single nanoparticles and shows how the radiative emission of a molecule can be further enhanced by engineering the geometry of a plasmonic structure. Through the use of rigorous 3D FDTD calculations, in conjunction with analytic guidance based on temporal coupled-mode (TCM) theory, we develop a design procedure for antennae assemblies that is useful both for general understanding of molecule-metal structure interaction and experimental efforts in plasmon-enhanced molecular spectroscopy. PMID:25321576

Meng, Xiang; Grote, Richard R; Dadap, Jerry I; Panoiu, Nicolae C; Osgood, Richard M

2014-09-01

78

Development of a Vital Fluorescent Staining Method for Monitoring Bacterial Transport in Subsurface Environments  

PubMed Central

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-105 CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well. PMID:11010903

Fuller, Mark E.; Streger, Sheryl H.; Rothmel, Randi K.; Mailloux, Brian J.; Hall, James A.; Onstott, Tullis C.; Fredrickson, James K.; Balkwill, David L.; DeFlaun, Mary F.

2000-01-01

79

Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments.  

PubMed

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-10(5) CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well. PMID:11010903

Fuller, M E; Streger, S H; Rothmel, R K; Mailloux, B J; Hall, J A; Onstott, T C; Fredrickson, J K; Balkwill, D L; DeFlaun, M F

2000-10-01

80

Treatment of Port-Wine Stains with Flash Lamp Pumped Pulsed Dye Laser on Indian Skin: A Six Year Study  

PubMed Central

Context: Port-wine stain (PWS) is one of the commonly encountered congenital cutaneous vascular lesions, with an equal sex distribution. Pulsed dye lasers (PDL) have revolutionized the treatment of both congential and acquired cutaneous vascular lesions. The pulsed dye lasers owing to its superior efficacy and safety profile have become the gold standard for the management of port-wine stains. Aims: To evaluate the efficacy and side effects of pulsed dye laser for the management of Port-wine stain on Indian skin. Materials and Methods: Seventy five patients of Fitzpatrick skin types IV&V with PWS underwent multiple treatments with PDL (V beam-Candela) over a period of six years at monthly intervals. Laser parameters were wavelength 595nm, spot sizes 7-10mm, fluence 6-12 j/cm2, pulse duration 0.45-10ms, along with cryogen cooling. Serial photographs were taken before and after every session. Clinical improvement scores of comparable photographs using a quartile grading (o=<20%, 1=21-40%, 2=41-60%, 3=61-80%, 4=>80%) were judged independently by two dermatologists after the series of treatment. Minimum number of treatments was 6 and maximum 17. They were followed up at six monthly intervals to observe re darkening of PWS. Results: No patient showed total clearance.Grade3 improvement was observed in 70 % of children and 50% of adults after 8-10 sessions. Children showed better and faster response than adults. Thirty percent of patients developed post inflammatory hyper pigmentation which resolved over a period of six to eight weeks. Two patients had superficial scarring due to stacking of pulses. None of the patients showed re darkening of PWS till now. Conclusion: Pulsed dye laser is an effective and safe treatment for port-wine stain in Indian skin. PMID:24761097

Thajudheen, Chandroth Ponnambath; Jyothy, Kannangath; Priyadarshini, Arul

2014-01-01

81

Cell-death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques.  

PubMed

Apoptosis, a genetically programmed cellular event leads to biochemical and morphological changes in cells. Alterations in DNA caused by several factors affect nucleus and ultimately the entire cell leading to compromised function of the organ and organism. DNA, a master regulator of the cellular events, is an important biomolecule with regards to cell growth, cell death, cell migration and cell differentiation. It is therefore imperative to develop the staining techniques that may lead to visualize the changes in nucleus where DNA is housed, to comprehend the cellular pathophysiology. Over the years a number of nuclear staining techniques such as propidium iodide, Hoechst-33342, 4', 6-diamidino-2-phenylindole (DAPI), Acridine orange-Ethidium bromide staining, among others have been developed to assess the changes in DNA. Some nonnuclear staining techniques such as Annexin-V staining, which although does not stain DNA, but helps to identify the events that result from DNA alteration and leads to initiation of apoptotic cell death. In this review, we have briefly discussed some of the most commonly used fluorescent and nonfluorescent staining techniques that identify apoptotic changes in cell, DNA and the nucleus. These techniques help in differentiating several cellular and nuclear phenotypes that result from DNA damage and have been identified as specific to necrosis or early and late apoptosis as well as scores of other nuclear deformities occurring inside the cells. PMID:24831993

Atale, N; Gupta, S; Yadav, U C S; Rani, V

2014-07-01

82

Toward ultra-stable fluorescent dyes for single-molecule spectroscopy  

NASA Astrophysics Data System (ADS)

The wide-spread use of fluorescent dyes in molecular diagnostics and fluorescence microscopy together with new developments such as single-molecule fluorescence spectroscopy provide researchers from various disciplines with an ever expanding toolbox. Single-molecule fluorescence spectroscopy relies to a large extent on extraordinary bright and photostable organic fluorescent dyes such as rhodamine- or cyanine- derivatives. While in the last decade singlemolecule equipment and methodology have significantly advanced and in some cases reached theoretical limits (e.g. detectors approaching unity quantum yields), instable emission ("blinking") and photobleaching become more and more the bottleneck of further development and spreading of single-molecule fluorescence studies. In recent years, agents and recipes have been developed to increase the photostability of conventional fluorescent dyes. Here, we investigate some of these strategies at the single-molecule level. In particular, we focus on the dye selection criteria for multi-color applications. We investigate fluorescent dyes from the rhodamine, carborhodamine, cyanine, and oxazine family and show that within one dye class the photophysical properties are very similar but that dyes from different classes show strikingly different properties. These findings facilitate dye selection and provide improved chemical environment for demanding fluorescence microscopic applications.

Kasper, Robert; Heilemann, Mike; Tinnefeld, Philip; Sauer, Markus

2007-07-01

83

pH-Sensitive Fluorescent Dyes: Are They Really pH-Sensitive in Cells?  

PubMed Central

Chemically synthesized near-infrared (NIR) aza-BODIPY dyes displayed OFF/ON fluorescence at acidic pH (pKa = 6.2-6.6) through the suppression of photoinduced electron transfer (PET) and/or internal charge transfer (ICT) process. The apparent pKas of the dyes were shifted well above physiological pH in hydrophobic microenvironment, which led to “turned-on” fluorescence in micelles and liposomes at neutral and basic pH. Bovine serum albumin (BSA) also activated the fluorescence, though to a much less extent. When these small molecular dyes entered cells, instead of being fluorescent only in acidic organelles, the whole cytoplasm exhibited fluorescence, with signal/background ratio as high as ?10 in no-wash live cell imaging. The dye 1 labeled cells remained highly fluorescent even after 3 days. Moreover, slight variations of the dye structure resulted in significantly different intracellular fluorescence behaviors, possibly due to their different cellular uptake and intracellular activation capabilities. After separation of cellular components, the fraction of plasma membrane and endoplasmic reticulum (ER) showed the highest fluorescence, further confirming the fluorescence activation by membrane structures. The fluorescence intensity of these dyes at different intracellular pH (6.80 and 8.00) did not differ significantly, indicating that intracellular pH did not play a critical role. Altogether, we showed here for the first time that the fluorescence of pH-sensitive aza-BODIPY dyes were switched intracellularly not by acidic pH, but by intracellular membranes (and proteins as well). The excellent membrane permeability, ultra high fluorescence contrast ratio, persistent fluorescent signal, and minimum biological interference of dye 1 make it an ideal choice for live cell imaging and in vivo cell tracking. These findings also imply that the intracellular fluorescent properties of pH-sensitive dyes should be carefully examined before used as pH indicators. PMID:23464828

Zhang, Xiao-Xiang; Wang, Zhe; Yue, Xuyi; Ma, Ying; Kiesewetter, Dale O.; Chen, Xiaoyuan

2013-01-01

84

Microarray Analysis of Port Wine Stains Before and After Pulsed Dye Laser Treatment  

PubMed Central

Background and Objectives Neither the pathogenesis of port wine stain (PWS) birthmarks nor tissue effects of pulsed dye laser (PDL) treatment of these lesions is fully understood. There are few published reports utilizing gene expression analysis in human PWS skin. We aim to compare gene expression in PWS before and after PDL, using DNA microarrays that represent most, if not all, human genes to obtain comprehensive molecular profiles of PWS lesions and PDL-associated tissue effects. Materials and Methods Five human subjects had PDL treatment of their PWS. One week later, three biopsies were taken from each subject: normal skin (N); untreated PWS (PWS); PWS post-PDL (PWS + PDL). Samples included two lower extremity lesions, two facial lesions, and one facial nodule. High-quality total RNA isolated from skin biopsies was processed and applied to Affymetrix Human gene 1.0ST microarrays for gene expression analysis. We performed a 16 pair-wise comparison identifying either up- or down-regulated genes between N versus PWS and PWS versus PWS + PDL for four of the donor samples. The PWS nodule (nPWS) was analyzed separately. Results There was significant variation in gene expression profiles between individuals. By doing pair-wise comparisons between samples taken from the same donor, we were able to identify genes that may participate in the formation of PWS lesions and PDL tissue effects. Genes associated with immune, epidermal, and lipid metabolism were up-regulated in PWS skin. The nPWS exhibited more profound differences in gene expression than the rest of the samples, with significant differential expression of genes associated with angiogenesis, tumorigenesis, and inflammation. Conclusion In summary, gene expression profiles from N, PWS, and PWS + PDL demonstrated significant variation within samples from the same donor and between donors. By doing pair-wise comparisons between samples taken from the same donor and comparing these results between donors, we were able to identify genes that may participate in formation of PWS and PDL effects. Our preliminary results indicate changes in gene expression of angiogenesis-related genes, suggesting that dysregulation of angiogenic signals and/or components may contribute to PWS pathology. PMID:23440713

Laquer, Vivian T.; Hevezi, Peter A.; Albrecht, Huguette; Chen, Tina S.; Zlotnik, Albert; Kelly, Kristen M.

2014-01-01

85

Identification of human hair stained with oxidation hair dyes by gas chromatographic-mass spectrometric analysis.  

PubMed

This paper describes the gas chromatographic-mass spectrometric (GCMS) analysis of oxidation hair dyes from human hair. Diamines from the dyes were directly extracted from the hair in basic solution and aminophenols were extracted after neutralization. Both extracts were derivatised with trifluoroacetic anhydride and analysed by GCMS. Five components of oxidation hair dyes namely, p-phenylenediamine, toluene-2,5-diamine, o-aminophenol, m-aminophenol and p-aminophenol were clearly identified, whilst no other compounds originating from the hair dyes were detected. The presence and relative amounts of these dye components from hair extracts may assist in the discrimination of human hair especially in cases involving forensic science. PMID:1783337

Tanada, N; Kageura, M; Hara, K; Hieda, Y; Takamoto, M; Kashimura, S

1991-12-01

86

Detection of early interproximal caries in vitro using laser fluorescence, dye-enhanced laser fluorescence and direct visual examination.  

PubMed

This in vitro study evaluated the use of laser fluorescence (LF) for the detection of early interproximal carious lesions and whether the detection could be enhanced using a fluorescent dye (DELF). Direct visual examination (DV) was used for comparison. Eighty extracted teeth were used, arranged in 20 blocks, each block having 2 premolars and 2 molars, lined up in a simulated sextant situation. After cleaning with a microabrasion kit, a subcontact window on half of the surfaces (60) was exposed to Carbopol white-spot solution for 5 days. The teeth were remounted in stone and examined by three independent examiners. For LF and DELF an argon laser was used (mixed wavelength of 488 and 514 nm) viewed through glasses (excluding wavelength <520 nm). For DELF a sodium fluorescein dye (0. 075%) was applied before examination. A clinical examination light was used for DV. The approximal surfaces were scored for lesion presence or absence. To verify lesion presence, the subcontact area was cut perpendicularly to the surface, stained with rhodamine B, and images were taken using a confocal microscope. The images were analyzed using a histogram program for lesion depth and image area. Lesions were present in 62 out of 120 approximal surfaces, with an average depth of 60 microm (range 17-190 microm). Sensitivity ranges for LF, DELF and DV were 56-74, 61-79 and 58-74%, and specificity ranges 67-78, 86-98 and 83-97%, respectively. With this model DELF compared favorably with DV and LF in sensitivity, but specificity was better for DELF and DV than for LF. PMID:10207199

Eggertsson, H; Analoui, M; van der Veen, M; González-Cabezas, C; Eckert, G; Stookey, G

1999-01-01

87

Diolistics: incorporating fluorescent dyes into biological samples using a gene gun  

PubMed Central

The hand-held gene gun provides a rapid and efficient method of incorporating fluorescent dyes into cells, a technique that is becoming known as diolistics. Transporting fluorescent dyes into cells has, in the past, used predominantly injection or chemical methods. The use of the gene gun, combined with the new generation of fluorescent dyes, circumvents some of the problems of using these methods and also enables the study of cells that have proved difficult traditionally to transfect (e.g. those deep in tissues and/or terminally differentiated); in addition, the use of ion- or metabolite-sensitive dyes provides a route to study cellular mechanisms. Diolistics is also ideal for loading cells with optical nanosensors – nanometre-sized sensors linked to fluorescent probes. Here, we discuss the theoretical considerations of using diolistics, the advantages compared with other methods of inserting dyes into cells and the current uses of the technique, with particular consideration of nanosensors. PMID:17945370

O’Brien, John A.; Lummis, Sarah C.R.

2007-01-01

88

Fluorescence of styryl dyes-DNA complexes induced by single- and two-photon excitation.  

PubMed

The series of novel monomer and homodimer styryl dyes based on (p-dimethylaminostyryl) benzothiazolium residues were synthesized and studied as possible fluorescent probes for nucleic acids detection. Spectral-luminescent and spectral-photometric properties of obtained dyes in the unbound state and in DNA presence were studied. Fluorescence emission induced by two-photon excitation of dye-DNA complexes in aqueous buffer solution was registered. Two-photon absorption cross section values of the studied dyes in DNA presence were evaluated. PMID:17031571

Tokar, V P; Losytskyy, M Yu; Kovalska, V B; Kryvorotenko, D V; Balanda, A O; Prokopets, V M; Galak, M P; Dmytruk, I M; Yashchuk, V M; Yarmoluk, S M

2006-11-01

89

Identification Of Natural Dyes On Archaeological Textile Objects Using Laser Induced Fluorescent Technique  

SciTech Connect

This study aims to evaluate the use of Laser Fluorescent as a non-destructive technique for identification of natural dyes on archaeological textile objects. In this study wool textile samples were dyed with 10 natural dyes such as cochineal, cutch, henna, indigo, Lac, madder, safflower, saffron, sumac and turmeric. These dyes common present on archaeological textile objects to be used as standard dyed textile samples. These selected natural dyes will be used as known references that can be used a guide to identify unknown archaeological dyes. The dyed textile samples were investigated with laser radiation in different wavelengths to detect the best wavelengths for identification each dye. This study confirms that Laser Florescent is very useful and a rapid technique can be used as a non-destructive technique for identification of natural dyes on archaeological textile objects. The results obtained with this study can be a guide for all conservators in identification of natural organic dyes on archaeological textile objects.

Abdel-Kareem, O. [Conservation Department, Faculty of Archaeology, Cairo University (Egypt); Eltokhy, A.; Harith, M. A. [National Institute of Laser Enhanced Science, Cairo University (Egypt)

2011-09-22

90

Identification Of Natural Dyes On Archaeological Textile Objects Using Laser Induced Fluorescent Technique  

NASA Astrophysics Data System (ADS)

This study aims to evaluate the use of Laser Fluorescent as a non-destructive technique for identification of natural dyes on archaeological textile objects. In this study wool textile samples were dyed with 10 natural dyes such as cochineal, cutch, henna, indigo, Lac, madder, safflower, saffron, sumac and turmeric. These dyes common present on archaeological textile objects to be used as standard dyed textile samples. These selected natural dyes will be used as known references that can be used a guide to identify unknown archaeological dyes. The dyed textile samples were investigated with laser radiation in different wavelengths to detect the best wavelengths for identification each dye. This study confirms that Laser Florescent is very useful and a rapid technique can be used as a non-destructive technique for identification of natural dyes on archaeological textile objects. The results obtained with this study can be a guide for all conservators in identification of natural organic dyes on archaeological textile objects.

Abdel-Kareem, O.; Eltokhy, A.; Harith, M. A.

2011-09-01

91

Highly Water-soluble Neutral BODIPY Dyes with Controllable Fluorescence Quantum Yields  

PubMed Central

series of novel highly water-soluble neutral BODIPY dyes have been obtained by functionalization of BODIPY dyes with branched oligo(ethylene glycol) methyl ether groups at positions 8, 2 and 6 or 4 and 4?. Use of an ortho-substituent group of branched oligo(ethylene glycol)methyl ether on the meso-phenyl ring of BODIPY dyes, and replacement of the fluorine atoms of BODIPY dyes at positions 4 and 4? with methyloxy or ethynyl subunits significantly enhances fluorescence quantum yields of BODIPY dyes. PMID:21175151

Zhu, Shilei; Zhang, Jingtuo; Vegesna, Giri; Luo, Fen-Tair; Green, Sarah

2011-01-01

92

Estrogen receptor-targeted optical imaging of breast cancer cells with near-infrared fluorescent dye  

NASA Astrophysics Data System (ADS)

Molecular imaging provides the in vivo characterization of cellular molecular events involved in normal and pathologic processes. With the advent of optical molecular imaging, specific molecules, proteins and genes may be tagged with a luminescent reporter and visualized in small animals. This powerful new tool has pushed in vivo optical imaging to the forefront as it allows for direct determination of drug bio-distribution and uptake kinetics as well as an indicator of biochemical activity and drug efficacy. Although optical imaging encompasses diverse techniques and makes use of various wavelengths of light, a great deal of excitement in molecular research lies in the use of tomographic and fluorescence techniques to image living tissues with near-infrared (NIR) light. Nonionizing, noninvasive near-infrared optical imaging has great potential to become promising alternative for breast cancer detection. Fluorescence spectroscopy studies of human tissue suggest that a variety of lesions show distinct fluorescence spectra compared to those of normal tissue. It has also been shown that exogenous dyes exhibit selective uptake in neoplastic lesions and may offer the best contrast for optical imaging. Use of exogenous agents would provide fluorescent markers, which could serve to detect embedded tumors in the breast. In particular, the ability to monitor the fluorescent yield and lifetime may also enable biochemical specificity if the fluorophore is sensitive to a specific metabolite, such as oxygen. As a first step, we have synthesized and characterized one such NIR fluorescent dye conjugate, which could potentially be used to detect estrogen receptors (ER)[2] . The conjugate was synthesized by ester formation between 17-? estradiol and a hydrophilic derivative of indocyanine green (ICG) cyanine dye, bis-1, 1-(4-sulfobutyl) indotricarbocyanine-5- carboxylic acid, sodium salt. The ester formed was found to have an extra binding ability with the receptor cites as compared to ICG, which was established by the partition coefficient studies. The replacement of the sodium ion in the ester by a larger glucosammonium ion was found to enhance the hydrophilicity and reduce the toxic effect on the cell lines. The excitation and emission peaks for the conjugate were recorded in the NIR region as 750nm and 788nm respectively. The ester was found nontoxic on adenocarcinoma breast cancer cell lines MCF-7/MDA-MB-231. Specific binding and endocytosis of the estrogen-labeled conjugate was studied on the MCF-7 (ER positive) and MDA-MB-231 (ER negative). Conjugate staining of MCF-7 cells showed ~ 4-fold increase in signal intensity compared to MDA-MB- 231. Further, estrogen molecules were found to be specifically localized to the nuclear region of MCF-7 cells, whereas MDA-MB-231 showed plasma membrane staining. This technique offers the potential of noninvasive detection of hormone receptor status in breast cancer cells and would help in decreasing the load of unnecessary biopsies. Here, we have reported the progress made in the development of a novel NIR external contrast agent and the work is in progress to use this conjugate for the molecular based, diagnostic imaging of breast cancer.

Jose, Iven; Deodhar, Kodand; Chiplunkar, Shuba V.; Patkar, Meena

2010-02-01

93

Rates of Benthic Protozoan Grazing on Free and Attached Sediment Bacteria Measured with Fluorescently Stained Sediment  

PubMed Central

In order to determine the importance of benthic protozoa as consumers of bacteria, grazing rates have been measured by using monodispersed fluorescently labeled bacteria (FLB). However, high percentages of nongrazing benthic protists are reported in the literature. These are related to serious problems of the monodispersed FLB method. We describe a new method using 5-(4,6-dichlorotriazin-2-yl)-aminofluorescein (DTAF)-stained sediment to measure in situ bacterivory by benthic protists. This method is compared with the monodispersed FLB technique. Our estimates of benthic bacterivory range from 61 to 73 bacteria protist-1 h-1 and are about twofold higher than the results of the monodispersed FLB method. The number of nongrazing protists after incubation for 15 min with DTAF-stained sediment is in agreement with theoretical expectation. We also tested the relative affinity for FLB of protists and discuss the results with respect to a grazing model. PMID:16349315

Starink, Mathieu; Krylova, Irina N.; Bär-Gilissen, Marie-José; Bak, Rolf P. M.; Cappenberg, Thomas E.

1994-01-01

94

A SERIES OF NEW DYES FOR CENTRAL NERVOUS SYSTEM SECTIONS STAINING. NOTE 1  

Microsoft Academic Search

The new diazo dye 5-(4-hydroxy-3-((4-(thiazol-2-yl sulfamoyl)phenyl) carbamoyl) phenyl) azo-2-(2-(4-(4-hydroxy-3-((4-(thiazol-2-ylsulfamoyl) phenyl) carbamoyl) phenyl) azo-2-sulfo-phenyl)vinyl)benzenesulfonic acid (called SINT 7) were obtained by synthesis using 4, 4'-diaminostilbene 2,2'-disulfonic acid (called AASS) and 2- hydroxy-N-(4-(thiazol-2-yl sulfamoyl) phenyl) benzamide. The diazo component (AASS) and the coupling component were likewise obtained by synthesis. (1). A similar diazo dye has not been prepared before, probably due to

RODICA MEHEDIN?I

2009-01-01

95

Staining diatoms with rhodamine dyes: control of emission colour in photonic biocomposites  

PubMed Central

The incorporation of rhodamine dyes in the cell wall of diatoms Coscinodiscus granii and Coscinodiscus wailesii for the production of luminescent hybrid nanostructures is investigated. By systematic variation of the substitution pattern of the rhodamine core, we found that carbonic acids are considerably better suited than esters because of their physiological compatibility. The amino substitution pattern that controls the optical properties of the chromophore has no critical influence on dye uptake and incorporation, thus a variety of biocomposites with different emission maxima can be prepared. Applications in biomineralization studies as well as in materials science are envisioned. PMID:21865248

Kucki, Melanie; Fuhrmann-Lieker, Thomas

2012-01-01

96

Macrophages possess probenecid-inhibitable organic anion transporters that remove fluorescent dyes from the cytoplasmic matrix  

Microsoft Academic Search

We introduced several membrane- impermeant fluorescent dyes, including Lucifer Yellow, carboxyfluorescein, and fura-2, into the cytoplasmic matrix of J774 cells and thioglycollate-elicited mouse peritoneal macrophages by ATP permeabilization of the plasma membrane and observed the subsequent fate of these dyes. The dyes did not remain within the cytoplasmic matrix; instead they were sequestered within phase-lucent cytoplasmic vacuoles and released into

Thomas H. Steinberg; Alan S. Newman; Joel A. Swanson; Samuel C. Silverstein

1987-01-01

97

Local delivery of fluorescent dye for fiber-optics confocal microscopy of the living heart  

PubMed Central

Fiber-optics confocal microscopy (FCM) is an emerging imaging technology with various applications in basic research and clinical diagnosis. FCM allows for real-time in situ microscopy of tissue at sub-cellular scale. Recently FCM has been investigated for cardiac imaging, in particular, for discrimination of cardiac tissue during pediatric open-heart surgery. FCM relies on fluorescent dyes. The current clinical approach of dye delivery is based on systemic injection, which is associated with high dye consumption, and adverse clinical events. In this study, we investigated approaches for local dye delivery during FCM imaging based on dye carriers attached to the imaging probe. Using three-dimensional confocal microscopy, automated bench tests, and FCM imaging we quantitatively characterized dye release of carriers composed of open-pore foam only and foam loaded with agarose hydrogel. In addition, we compared local dye delivery with a model of systemic dye delivery in the isolated perfused rodent heart. We measured the signal-to-noise ratio (SNR) of images acquired in various regions of the heart. Our evaluations showed that foam-agarose dye carriers exhibited a prolonged dye release vs. foam-only carriers. Foam-agarose dye carriers allowed reliable imaging of 5–9 lines, which is comparable to 4–8 min of continuous dye release. Our study in the living heart revealed that the SNR of FCM images using local and systemic dye delivery is not different. However, we observed differences in the imaged tissue microstructure with the two approaches. Structural features characteristic of microvasculature were solely observed for systemic dye delivery. Our findings suggest that local dye delivery approach for FCM imaging constitutes an important alternative to systemic dye delivery. We suggest that the approach for local dye delivery will facilitate clinical translation of FCM, for instance, for FCM imaging during pediatric heart surgery. PMID:25309455

Huang, Chao; Kaza, Aditya K.; Hitchcock, Robert W.; Sachse, Frank B.

2014-01-01

98

Histologic responses of port-wine stains treated by argon, carbon dioxide, and tunable dye lasers. A preliminary report.  

PubMed

Although the blue-green argon laser light has been used successfully to treat port-wine stains (PWSs) for many years, it produces substantial epidermal damage. We have previously shown in normal human skin that pulsed yellow tunable dye lasers (577-nm wavelength) can cause highly selective damage to cutaneous microvessels with minimal injury to the overlying epidermis. Pulsed tunable dye lasers also produce selective vascular injury in the abnormal vessels of PWSs, with clinically apparent lightening of the lesions. Both epidermal injury and fibrosis are less severe with this laser treatment than with argon and carbon dioxide laser treatments. Clinical and histologic responses of PWSs treated by argon, CO2, and pulsed yellow dye lasers were compared and followed up for one month in three patients. Although larger and longer-term clinical trials are necessary to fully evaluate this new treatment modality, it appears that pulsed yellow laser radiation offers a more selective, less traumatic, and probably superior form of treatment for PWSs. PMID:3090945

Tan, O T; Carney, J M; Margolis, R; Seki, Y; Boll, J; Anderson, R R; Parrish, J A

1986-09-01

99

Phthalocyanine dye as an extremely photostable and highly fluorescent near-infrared labeling reagent  

NASA Astrophysics Data System (ADS)

Current organic fluorophores used as labeling reagents for biomolecule conjugation have significant limitations in photostability. This compromises their performance in applications that require a photostable fluorescent reporting group. For example, in molecular imaging and single molecule microscopy, photostable fluorescent labels are important for observing and tracking individual molecular events over extended period of time. We report in this paper an extremely photostable and highly fluorescent phthalocyanine dye, IRDye TM 700DX, as a near-infrared fluorescence labeling reagent to conjugate with biomolecules. This novel water-soluble silicon phthalocyanine dye has an isomericly pure chemical structure. The dye is about 45 to 128 times more photostable than current near-IR fluorophores, e.g. Alexa Fluor"R"680, Cy TM 5.5, Cy TM 7 and IRDye TM 800CW dyes; and about 27 times more photostable than tetramethylrhodamine (TMR), one of the most photostable organic dyes. This dye also meets all the other stringent requirements as an ideal fluorophore for biomolecules labeling such as excellent water solubility, no aggregation in high ionic strength buffer, large extinction coefficient and high fluorescent quantum yield. Antibodies conjugated with IRDye TM 700DX at high D/P ratio exist as monomeric species in high ionic buffer and have bright fluorescence. The IRDye TM 700DX conjugated antibodies generate sensitive, highly specific detection with very low background in Western blot and cytoblot assays.

Peng, Xinzhan; Draney, Daniel R.; Volcheck, William M.; Bashford, Gregory R.; Lamb, Donald T.; Grone, Daniel L.; Zhang, Yonghong; Johnson, Craig M.

2006-02-01

100

Confocal microscopy of multiple-stained biological specimens using fluorescence lifetimes  

NASA Astrophysics Data System (ADS)

We here report on using fluorescence life times recordings in confocal microscopy to detect individual fluorophores in multiple stained tissue. Using indirect fluorescence immunohistochemistry with secondary antisera conjugated to the fluorophores Lissamine Rhodamine (LRSC), Texas Red or Cyanine 3.18 (Cy-3); one, two or three epitopes were labelled in tissues from rat spinal cord and dorsal root ganglia. The tissue sections were examined and analyzed in a beam-scanning confocal microscope equipped with devices for fluorescence lifetime measurements. The results show that fluorophore life-times can be used to separate two or more fluorophores in individual axon terminals, provided that the fluorophore labelling is strong enough and the life-times of the deployed compounds are sufficiently separate. Thus, the presence of Cy-3, LRSC and Texas Red, as well as mixtures of these compounds could be detected in individual tissue profiles. Contribution by tissue autofluorescence was unmistakably identified by its complex multiexponential emission decay. We also show that fluorescence lifetimes differs between antiserum-conjugates for one and the same fluorophore, and the possibility to use fluorophore life-times to probe chemical environmental changes in situ is discussed. The implementation of a life-time recording device in a confocal microscope has the advantage of providing a good spatial resolution. Furthermore, by deploying fluorophores emitting within the same wavelength band, problems due to chromatic errors will be completely avoided.

Brismar, Hjalmar; Ulfhake, Brun

1995-03-01

101

Fluorescence properties and staining behavior of monodansylpentane, a structural homologue of the lysosomotropic agent monodansylcadaverine.  

PubMed

We have recently shown that monodansylcadaverine labels autophagic vacuoles. Analysis of the mechanism underlying the labeling revealed that monodansylcadaverine acts as a lysosomotropic agent, being concentrated into acidic compartments by an ion-trapping mechanism, and as a solvent polarity probe, increasing its relative fluorescence intensity by interacting with membrane lipids that are highly concentrated in the autophagic vacuoles. In this study, we synthesized three structurally related derivatives of monodansylcadaverine, replacing the primary amino group of monodansylcadaverine with a neutral (dansylamylamine; MDH), a polar (dansylaminopentanol; MDOH), or an acidic group (dansylaminovaleric acid; MDA), to replace the lysosomotropic character of the marker. Whereas MDH showed a specific staining of autophagic vacuoles, the polar and acidic derivatives did not show any staining. We further demonstrate that the MDH staining of autophagic vacuoles is independent on the acidic pH and thus on an ion-trapping mechanism, but it still shows the same preferences for autophagic membrane lipids as monodansylcadaverine. We propose that MDH can specifically interact with lamellar bodies of the autophagic type as a solvent polarity probe. Therefore, dansylated aminopentane can be used as a specific marker for autophagic vacuoles in vivo and in fixed cells.(J Histochem Cytochem 49:177-185, 2001) PMID:11156686

Niemann, A; Baltes, J; Elsässer, H P

2001-02-01

102

Dependence of detected intensity of fluorescence of dye on optical fiber tapered tip diameter  

NASA Astrophysics Data System (ADS)

The measurement of pH in small objects (cells, drops of liquid etc.) using optical fluorescence-based sensors on optical fiber tapers is one of the most widely used optical techniques. In these sensors the diameter of the taper can play important role for collecting fluorescence from tested samples. This paper presents results of experimental measurements of fluorescence intensity of dye sensitive to pH in a solution that is excited by a blue laser. The fluorescence of the dye is collected by a taper tip. The fiber tips were prepared from a graded-index fiber with a core diameter of 50 ?m. Measurements with taper tips of different diameters have allowed us to estimated a limited tip diameter necessary for collecting any fluorescence form the dye solution on a level of about 5?m.

Pospisilova, M.; Aubrecht, J.; Martan, T.; Mrazek, J.; Kasik, I.; Matejec, V.

2011-05-01

103

Polylysine crosslinked AIE dye based fluorescent organic nanoparticles for biological imaging applications.  

PubMed

Fluorescent organic nanoparticles based on aggregation induced emission dyes are fabricated through a ring-opening reaction using polylysine as the linker. The fluorescent organic nanoparticles obtained are characterized by a series of techniques including UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier Transform infrared spectroscopy, and transmission electron microscopy. A biocompatibility evaluation and the cell uptake behavior of the fluorescent organic nanoparticles are further investigated to evaluate their potential biomedical applications. It is demonstrated that these fluorescent organic nanoparticles can be obtained at room temperature in an air atmosphere without the need for catalyst or initiator. Furthermore, these crosslinked aggregation induced emission dye based fluorescent organic nanoparticles show uniform morphology, strong red fluorescence, high water dispersability, and excellent biocompatibility, making them promising candidates for various biomedical applications. PMID:24854875

Liu, Meiying; Zhang, Xiqi; Yang, Bin; Liu, Liangji; Deng, Fengjie; Zhang, Xiaoyong; Wei, Yen

2014-09-01

104

Method and apparatus for staining immobilized nucleic acids  

DOEpatents

A method for staining immobilized nucleic acids includes the steps of affixing DNA probes to a solid substrate, moving target DNA material into proximity with the DNA probes, whereby the target DNA hybridized with specific ones of the DNA probes, and moving a fluorescent dye into proximity with the hybridized target DNA, whereby the fluorescent dye binds to the hybridized DNA to enable subsequent detection of fluorescence.

Ramsey, J. Michael (Knoxville, TN); Foote, Robert S. (Oak Ridge, TN); Jacobson, Stephen C. (Knoxville, TN)

2000-01-01

105

Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications.  

PubMed Central

The synthesis, proof of structure, and the absorption and fluorescence properties of two new unsymmetrical cyanine dyes, thiazole orange dimer (TOTO; 1,1'-(4,4,7,7-tetramethyl-4,7- diazaundecamethylene)-bis-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiaz ole)-2- methylidene]-quinolinium tetraiodide) and oxazole yellow dimer (YOYO; an analogue of TOTO with a benzo-1,3-oxazole in place of the benzo-1,3-thiazole) are reported. TOTO and YOYO are virtually non-fluorescent in solution, but form highly fluorescent complexes with double-stranded DNA (dsDNA), up to a maximum dye to DNA bp ratio of 1:4, with greater than 1000-fold fluorescence enhancement. The dsDNA-TOTO (lambda max 513 nm; lambda maxF 532 nm) and dsDNA-YOYO (lambda max 489 nm; lambda maxF 509 nm) complexes are completely stable to electrophoresis on agarose and acrylamide gels. Mixtures of restriction fragments pre-labeled with ethidium dimer (EthD; lambda maxF 616 nm) and those pre-labeled with either TOTO or YOYO were separated by electrophoresis. Laser excitation at 488 nm and simultaneous confocal fluorescence detection at 620-750 nm (dsDNA-EthD emission) and 500-565 nm (dsDNA-TOTO or dsDNA-YOYO emission) allowed sensitive detection, quantitation, and accurate sizing of restriction fragments ranging from 600 to 24,000 bp. The limit of detection of dsDNA-TOTO and YOYO complexes with a laser-excited confocal fluorescence gel scanner for a band 5-mm wide on a 1-mm thick agarose gel was 4 picograms, about 500-fold lower than attainable by conventional staining with ethidium bromide. Images PMID:1614866

Rye, H S; Yue, S; Wemmer, D E; Quesada, M A; Haugland, R P; Mathies, R A; Glazer, A N

1992-01-01

106

A novel frequency method for quantitative analysis of fluorescence dye concentration by using series photodetector frequency circuit system  

Microsoft Academic Search

This study reported the frequency characteristics of series photodetector frequency circuit system for detection of fluorescence dye concentration. In the condition of the same fluorescence intensity, the series photodetector frequency circuit system with higher responsivity of photodetector had higher frequency shift. The 100MHz series photodetector frequency circuit system was applied to determine the fluorescence dye concentration of HEX by frequency

Yuh Ming Hsu; Chung Cheng Chang

2009-01-01

107

Aerial Imaging of Fluorescent Dye in the Near Shore DAVID B. CLARK  

E-print Network

measurements are costly and complex, and few fluorescent dye studies have been conducted in turbid nearshore waters. Passive aerial imaging of solar stimulated fluorescence offers a relatively simple, low) in the nearshore zone have not been described. Here we develop simple airborne-based optical methods for measuring

Boss, Emmanuel S.

108

Functional molecular lumino-materials to probe serum albumins: solid phase selective staining through noncovalent fluorescent labeling.  

PubMed

Selective staining of human serum albumin protein in gel electrophoresis over wide range of other protein(s) is extremely important because it contains more than 60% volume of serum fluid in human body. Given the nonexistence of suitable dye materials for selective staining of serum albumins in gel electrophoresis, we report a new class of easy synthesizable and low molecular weight staining agents based on 3-amino-N-alkyl-carbazole scaffold for selective staining of serum albumins in solid phase. A detailed structure-efficiency relationship (SER) study enabled us to develop two such potent functional molecular probes which stain both human and bovine serum albumin selectively in gel electrophoresis in the presence of other proteins and enzymes. The present gel staining process was found to be very simple and less time-consuming as compared to the conventional coomassie blue staining which in turn makes these probes a new class of serum albumin-specific staining materials in proteome research. Moreover, these molecular lumino-materials can detect serum albumins at subnanomolar level in the presence of broad spectrum of other proteins/enzymes in aqueous buffer (99.9% water, pH = 7.3) keeping the protein secondary structure intact. Our experimental and the docking simulation results show that these probes bind preferentially at 'binding site I' of both the serum proteins. PMID:24926791

Dey, Gourab; Gupta, Abhishek; Mukherjee, Trinetra; Gaur, Pankaj; Chaudhary, Abhishek; Mukhopadhyay, Subhra Kanti; Nandi, Chayan K; Ghosh, Subrata

2014-07-01

109

Use of fluorescent NIR dyes in silica nanoparticles and as enzyme substrates in bioanalytical applications  

NASA Astrophysics Data System (ADS)

Near-Infrared (NIR) absorbing carbocyanine dyes have been increasingly used in analytical, biological and medical fields as they can be useful for developing bioanalytical and biomedical methods. The utilization of the NIR spectral region (650-900 nm) is advantageous and is due to the inherently lower background interference and the high molar absorptivities of NIR chromophores. NIR dyes typically have relatively lower fluorescent quantum yield as compared to visible fluorophores, but much higher molar absorptivities which more than compensates for the lower quantum yields regarding detection limits. Fluorescence intensity of NIR dyes significantly increases by enclosing several dye molecules in silica nanoparticles. Self quenching may become a problem for carbocyanines at such high concentrations that may be present in the silica nanoparticles. Dyes that have large Stokes' shift can significantly decrease this problem. Increased Stokes' shift for carbocyanines dyes can be achieved by substituting meso position halogens with a linker containing aliphatic or aromatic amino moiety which also serves as a covalent linker for attaching the dye molecule to the nanoparticle backbone. The primary applications of these particles are for bright fluorescent labels to be used in bioanalytical applications such as immunochemistry, flow cytometry, etc. This work also discusses the use of NIR dyes as enzyme substrates. NIR dyes can be used as enzyme substrates and hence for characterization of enzyme activity. The well characterized alkenesulfonate monooxygenase enzyme was chosen for these studies. Carbocyanines containing alkylsulfonate moieties do not exhibit significant fluorescence change upon binding to biomolecules however otherwise identical NIR dye analogs that contain alkylaldehyde moiety at the same position do exhibit changes which can be utilized for characterization of alkenesulfonate monooxygenase enzyme activity using near infrared dyes as substrates. In this study a new class of sulfonated penta- and heptamethine dyes were used as substrates in vitro utilizing a photo-reduced riboflavin mononucleotide (FMN) with a glucose/ glucose-oxygenase oxygen scavenging system. Laser Induced Fluorescence (LIF) detected CZE was utilized to detect the sulfonated and de-sulfonated carbocyanines. The lower fluorescence quantum yield of the less water soluble alkylaldehyde analogs was detected and enzyme activity was characterized.

Patonay, Gabor; Chapman, Gala; Beckford, Garfield; Henary, Maged; Ellis, Holly

2014-03-01

110

Characterizing the Interaction Between DNA and GelRed Fluorescent Stain  

E-print Network

We have performed single molecule stretching experiments and dynamic light scattering (DLS) in order to characterize the interaction between the DNA molecule and the fluorescent stain GelRed. The results from single molecule stretching show that the persistence length of the DNA-GelRed complexes increases as the ligand concentration increases up to some critical concentration, then decreasing for higher concentrations. The contour length of the complexes, on the other hand, increases monotonically as a function of GelRed concentration, suggesting that intercalation is the main binding mechanism. In order to characterize the phys- ical chemistry of the interaction, we use the McGhee-von Hippel binding isotherm to extract the physicochemical parameters of the interaction from the contour length data. The DLS experiments were performed to study the changes of the effective size of the DNA-GelRed complexes, measured by the hydrodynamic radius, as a function of ligand concentration. We found a qualitative agreemen...

Crisafuli, F A P; Rocha, M S

2014-01-01

111

Fluorescence of Dyes in Solutions with High Absorbance. Inner Filter Effect Correction  

PubMed Central

Fluorescence is a proven tool in all fields of knowledge, including biology and medicine. A significant obstacle in its use is the nonlinearity of the dependence of the fluorescence intensity on fluorophore concentration that is caused by the so-called primary inner filter effect. The existing methods for correcting the fluorescence intensity are hard to implement in practice; thus, it is generally considered best to use dilute solutions. We showed that correction must be performed always. Furthermore, high-concentration solutions (high absorbance) are inherent condition in studying of the photophysical properties of fluorescent dyes and the functionally significant interactions of biological macromolecules. We proposed an easy to use method to correct the experimentally recorded total fluorescence intensity and showed that informative component of fluorescence intensity numerically equals to the product of the absorbance and the fluorescence quantum yield of the object. It is shown that if dye molecules do not interact with each other and there is no reabsorption (as for NATA) and spectrofluorimeter provides the proportionality of the detected fluorescence intensity to the part of the absorbed light (that is possible for spectrofluorimeter with horizontal slits) then the dependence of experimentally detected total fluorescence intensity of the dye on its absorbance coincides with the calculated dependence and the correction factor for eliminating the primary inner filter effect can be calculated on the basis of solution absorbance. It was experimentally shown for NATA fluorescence in the wide range of absorbance (at least up to 60). For ATTO-425, which fluorescence and absorption spectra overlap, the elimination of the primary and secondary filter effects and additional spectral analysis allow to conclude that the most probable reason of the deviation of experimentally detected fluorescence intensity dependence on solution absorbance from the calculated dependence is the dye molecules self-quenching, which accompanies resonance radiationless excitation energy transfer. PMID:25072376

Fonin, Alexander V.; Sulatskaya, Anna I.; Kuznetsova, Irina M.; Turoverov, Konstantin K.

2014-01-01

112

Characterizing the interaction between DNA and GelRed fluorescent stain.  

PubMed

We have performed single-molecule stretching and dynamic light-scattering (DLS) experiments to characterize the interaction between the DNA molecule and the fluorescent stain GelRed. The results from single-molecule stretching show that the persistence length of DNA-GelRed complexes increases as the ligand concentration increases up to a critical concentration, then decreases for higher concentrations. The contour length of the complexes, on the other hand, increases monotonically as a function of GelRed concentration, suggesting that intercalation is the main binding mechanism. To characterize the physical chemistry of the interaction, we used the McGhee-von Hippel binding isotherm to extract physicochemical data for the interaction from the contour length data. Such analysis enabled us to conclude that the GelRed stain is, in fact, a bis-intercalator. In addition, DLS experiments were performed to study the changes of the effective size of the DNA-GelRed complexes, measured as the hydrodynamic radius, as a function of ligand concentration. We observed qualitative agreement between the results obtained from the two techniques by comparing the behavior of the hydrodynamics radius and the radius of gyration, because the latter quantity can be expressed as a function of mechanical properties determined from the stretching experiments. PMID:25391339

Crisafuli, F A P; Ramos, E B; Rocha, M S

2015-02-01

113

Green tea catechins quench the fluorescence of bacteria-conjugated Alexa fluor dyes.  

PubMed

Accumulating evidence suggests that Green tea polyphenolic catechins, especially the (-)-epigallocatechin gallate (EGCG), can be cross-linked to many proteins, and confer a wide range of anti-bacterial activities possibly by damaging microbial cytoplasmic lipids and proteins. At the doses that conferred protection against lethal polymicrobial infection (induced by cecal ligation and puncture), EGCG significantly reduced bacterial loads particularly in the liver and lung. To elucidate its bactericidal mechanisms, we determined whether EGCG affected the fluorescence intensities of bacteria-conjugated Alexa Fluor 488 or 594 dyes. When mixed with unconjugated Alexa Fluor 488 or 594 dyes, EGCG or analogs did not affect the fluorescence intensity of these dyes. In a sharp contrast, EGCG and some analogs (e.g., Catechin Gallate, CG), markedly reduced the fluorescence intensity of Gram-positive Staphylococcus aureus-conjugated Alexa 594 and Gram-negative Escherichia coli-conjugated Alexa 488. Interestingly, co-treatment with ethanol impaired the EGCG-mediated fluorescence quenching of the G(+) S. aureus, but not of the G(-) E. coli-conjugated Alexa Flour dyes. In light of the notion that Alexa Fluor dyes can be quenched by aromatic amino acids, it is plausible that EGCG exerts antimicrobial activities possibly by altering microbial protein conformations and functions. This possibility can now be explored by screening other fluorescence-quenching agents for possible antimicrobial activities. PMID:24011199

Zhao, Lin; Li, Wei; Zhu, Shu; Tsai, Sheena; Li, Jianhua; Tracey, Kevin J; Wang, Ping; Fan, Saijun; Sama, Andrew E; Wang, Haichao

2013-10-01

114

Photophysics of Laser Dye-Doped Polymer Membranes for Laser-Induced Fluorescence Photogrammetry  

NASA Technical Reports Server (NTRS)

Laser-induced fluorescence target generation in dye-doped polymer films has recently been introduced as a promising alternative to more traditional photogrammetric targeting techniques for surface profiling of highly transparent or reflective membrane structures. We investigate the photophysics of these dye-doped polymers to help determine their long-term durability and suitability for laser-induced fluorescence photogrammetric targeting. These investigations included experimental analysis of the fluorescence emission pattern, spectral content, temporal lifetime, linearity, and half-life. Results are presented that reveal an emission pattern wider than normal Lambertian diffuse surface scatter, a fluorescence time constant of 6.6 ns, a pump saturation level of approximately 20 micro J/mm(exp 2), and a useful lifetime of more than 300,000 measurements. Furthermore, two demonstrations of photogrammetric measurements by laser-induced fluorescence targeting are presented, showing agreement between photogrammetric and physically measured dimensions within the measurement scatter of 100 micron.

Dorrington, Adrian A.; Jones, Thomas W.; Danehy, Paul M.

2004-01-01

115

Diketopyrrolopyrrole: brilliant red pigment dye-based fluorescent probes and their applications.  

PubMed

The development of fluorescent probes for the detection of biologically relevant species is a burgeoning topic in the field of supramolecular chemistry. A number of available dyes such as rhodamine, coumarin, fluorescein, and cyanine have been employed in the design and synthesis of new fluorescent probes. However, diketopyrrolopyrrole (DPP) and its derivatives have a distinguished role in supramolecular chemistry for the design of fluorescent dyes. DPP dyes offer distinctive advantages relative to other organic dyes, including high fluorescence quantum yields and good light and thermal stability. Significant advancements have been made in the development of new fluorescent probes based on DPP in recent years as a result of tireless research efforts by the chemistry scientific community. In this tutorial review, we highlight the recent progress in the development of DPP-based fluorescent probes for the period spanning 2009 to the present time and the applications of these probes to recognition of biologically relevant species including anions, cations, reactive oxygen species, thiols, gases and other miscellaneous applications. This review is targeted toward providing the readers with deeper understanding for the future design of DPP-based fluorogenic probes for chemical and biological applications. PMID:25186723

Kaur, Matinder; Choi, Dong Hoon

2015-01-01

116

Dye distance mapping using waveguide evanescent field fluorescence microscopy and its application to cell biology.  

PubMed

Previous studies have measured the distance between cells and the substratum at sites of adhesion via the emission of a fluorescent dye and waveguide methods. Here, we demonstrate a novel approach to measure the position of fluorescent dyes above a waveguide surface in the 10-200 nm distance range throughout an entire area, yielding a 2D dye distance map or a 3D contour plot. The dye is located in a multilayered Langmuir Blodgett (LB) film or in the plasma membrane of a cell. Waveguide evanescent field fluorescence (WEFF) images obtained using two different waveguide modes are employed allowing, with a simple mathematical approach, the calculation of dye distance maps. Ultra-thin steps made using LB technology, adhesion distances and the bending of the plasma membrane between focal adhesions of osteoblastic cells are shown as examples. The errors are discussed. False color representation of a dye distance map with four osteoblasts. The inset represents an overexposed WEFF image of the same field of view. PMID:25401699

Fleissner, Frederik; Morawitz, Michael; Dixon, S Jeffrey; Langbein, Uwe; Mittler, Silvia

2014-11-17

117

Microfiberoptic Measurement of Extracellular Space Volume in Brain and Tumor Slices Based on Fluorescent Dye Partitioning  

PubMed Central

The fractional volume occupied by extracellular space in tissues, termed ?, is an important parameter of tissue architecture that affects cellular functions and drug delivery. We report a technically simple fluorescent dye partitioning method to measure ? in tissue slices based on microfiberoptic detection of dye fluorescence in tissue versus overlying solution. Microfiberoptic tip geometry and dyes were selected for ? determination from fluorescence intensity ratios, without the need to correct for illumination profile, light scattering/absorption, or dye binding. The method was validated experimentally using cell-embedded gels of specified ?-values and optical properties. In mouse brain slices, ? was strongly location-dependent, ranging from 0.16 in thalamus to 0.22 in brainstem, and was sensitive to cell volume changes. Aquaporin-4 water channel gene deletion caused significant extracellular space expansion, with ? = 0.181 ± 0.002 in cortex in wild-type mice and 0.211 ± 0.003 in Aquaporin-4 knockout mice. In slices of LLC1 cell tumors grown in mice to ?5 mm diameter, ? decreased remarkably from ?0.45 in superficial tumor to <0.25 in deeper (>100 ?m) tumor. Fluorescent dye partitioning with microfiberoptic detection permits rapid, accurate, and anisotropy-insensitive determination of ?-values in tissue slices. PMID:20713014

Zhang, Hua; Verkman, A.S.

2010-01-01

118

Fluorescent styryl dyes as probes for Na,K-ATPase reaction mechanism: significance of the charge of the hydrophilic moiety of RH dyes.  

PubMed

The fluorescence responses of a series of potential-sensitive styryl-based dyes (either zwitterionic RH160, RH421, di-4-ANEPPS, or positively charged RH795, RH414, RH461) to phosphorylation of Na,K-ATPase from ATP or inorganic phosphate, and ouabain binding to phospho- or dephosphoforms, have been characterized and compared in broken membrane preparations of the enzyme. Zwitterionic dyes were more sensitive to molecular events in the Na,K-ATPase reaction cycle than positively charged dyes, but the net charge did not affect the sensitivity of the dyes to a transmembrane electric field. The major part of the response of the zwitterionic dyes to formation of phosphoenzymes was due to a change in the quantum yield of fluorescence. Computer modeling of dyes with identical chromophore structure, and experimental characterization of their optical properties in bulk solvents, revealed two general trends: (1) the absorption maximum of the zwitterionic dye was blue-shifted with respect to the positively charged dye; (2) the quantum yield of the zwitterionic dye was higher and the fluorescence lifetime was longer than that for the positively charged dye. Spectral properties of the dyes in the membrane depended on the presence of Na,K-ATPase. We suggest, that (1) electrostatic interactions between the enzyme and the hydrophilic headgroup of the dye by changing the charge of hydrophilic moiety and thus modifying the net charge of the dye molecule cause both the spectral shifts and the changes in the quantum yield, and (2) interactions between the styryl dyes and the Na,K-ATPase depend on the conformational state of the enzyme. PMID:8527456

Fedosova, N U; Cornelius, F; Klodos, I

1995-12-26

119

Bacterial Viability and Antibiotic Susceptibility Testing with SYTOX Green Nucleic Acid Stain  

Microsoft Academic Search

A fluorescent nucleic acid stain that does not penetrate living cells was used to assess the integrity of the plasma membranes of bacteria. SYTOX Green nucleic acid stain is an unsymmetrical cyanine dye with three positive charges that is completely excluded from live eukaryotic and prokaryotic cells. Binding of SYTOX Green stain to nucleic acids resulted in a >500-fold enhancement

BRUCE L. ROTH; MARTIN POOT; STEPHEN T. YUE; PAUL J. MILLARD

1997-01-01

120

Photosensitive Fluorescent Dye Contributes to Phototoxicity and Inflammatory Responses of Dye-doped Silica NPs in Cells and Mice  

PubMed Central

Dye-doped fluorescent silica nanoparticles provide highly intense and photostable fluorescence signals. However, some dopant dye molecules are photosensitive. A widely-used photosensitive fluorescent dopant, RuBpy, was chosen to systematically investigate the phototoxicity of the dye-doped silica nanoparticles (NPs). We investigated cell viability, DNA damage, and Reactive Oxygen Species (ROS) levels in alveolar macrophages using the dye-doped NPs with or without irradiation. Our results showed that the RuBpy-doped silica NPs could induce significant amount of ROS, DNA damage, apoptosis and impaired proliferation in MH-S cells. In vivo studies in mice showed that RuBpy-doped silica NPs induced significant inflammatory cytokine production and lowered expression in signaling proteins such as ERK1/2 and NF-?B as well as increased lung injury determined by myeloperoxidase and lipid peroxidation. Strikingly, we also found that both RuBpy alone and NPs induced systemic signaling activation in the kidney compared to the liver and lung where showed highly selective signaling patterns, which is more pronounced than RuBpy-doped silica NPs. Moreover, we discovered a critical biomarker (e.g., HMGB1) for silica NPs-induced stress and toxicity and demonstrated differentially-regulated response patterns in various organs. Our results indicate for the first time that the RuBpy-doped silica NPs may impose less inflammatory responses but stronger thermotherapeutic effects on target cells in animals than naked NPs in a time- and dose-dependent manner. PMID:24578727

Zhao, Yang; Ye, Yan; Zhou, Xikun; Chen, Jiao; Jin, Yuihui; Hanson, Aaron; Zhao, Julia Xiaojun; Wu, Min

2014-01-01

121

A method for evaluating the use of fluorescent dyes to track proliferation in cell lines by dye dilution.  

PubMed

Labeling nonquiescent cells with carboxyfluorescein succinimidyl ester (CFSE)-like dyes gives rise to a population width exceeding the threshold for resolving division peaks by flow cytometry. Width is a function of biological heterogeneity plus extrinsic and intrinsic error sources associated with the measurement process. Optimal cytometer performance minimizes extrinsic error, but reducing intrinsic error to the point of facilitating peak resolution requires careful fluorochrome selection and fluorescent cell sorting. In this study, we labeled the Jurkat and A549 cell lines with CFSE, CellTraceViolet (CTV), and eFluor 670 proliferation dye (EPD) to test if we could resolve division peaks in culture after reducing the labeled input widths by cell sorting. Reanalysis of the sorted populations to ascertain the level of reduction achieved always led to widths exceeding the gated limits due to the contribution of errors. Measuring detector-specific extrinsic error by sorting uniform fluorescent particles with similar spectral properties to the tracking dyes allowed us to determine the intrinsic error for each dye and cell type using a simple mathematical approach. We found that cell intrinsic error ultimately dictated whether we could resolve division peaks, and that as this increased, the required sort gate width to resolve any division peaks decreased to the point whereby issues with yield made A549 unsuitable for this approach. Finally, attempts to improve yields by setting two concurrent sort gates on the fluorescence distribution enriched for cells in different stages of the cell cycle that had nonequivalent proliferative properties in culture and thus should be practiced with caution. PMID:24166880

Begum, Julfa; Day, William; Henderson, Carl; Purewal, Sukhveer; Cerveira, Joana; Summers, Huw; Rees, Paul; Davies, Derek; Filby, Andrew

2013-12-01

122

Testing the Fraunhofer line discriminator by sensing fluorescent dye  

NASA Technical Reports Server (NTRS)

The experimental Fraunhofer Line Discriminator (FLD) has detected increments of Rhodamine WT dye as small as 1 ppb in 1/2 meter depths. It can be inferred that increments considerably smaller than 1 ppb will be detectable in depths considerably greater than 1/2 meter. Turbidity of the water drastically reduces luminescence or even completely blocks the transmission of detectable luminescence to the FLD. Attenuation of light within the water by turbidity and by the dye itself are the major factors to be considered in interpreting FLD records and in relating luminescence coefficient to dye concentration. An airborne test in an H-19 helicopter established feasibility of operating the FLD from the aircraft power supply, and established that the rotor blades do not visibly affect the monitoring of incident solar radiation.

Stoertz, G. E.

1969-01-01

123

A double fluorescence staining protocol to determine the cross-sectional area of myofibers using image analysis  

NASA Technical Reports Server (NTRS)

A double fluorescence staining protocol was developed to facilitate computer based image analysis. Myofibers from experimentally treated (irradiated) and control growing turkey skeletal muscle were labeled with the anti-myosin antibody MF-20 and detected using fluorescein-5-isothiocyanate (FITC). Extracellular material was stained with concanavalin A (ConA)-Texas red. The cross-sectional area of the myofibers was determined by calculating the number of pixels (0.83 mu m(2)) overlying each myofiber after subtracting the ConA-Texas red image from the MF-20-FITC image for each region of interest. As expected, myofibers in the irradiated muscle were smaller (P < 0.05) than those in the non-irradiated muscle. This double fluorescence staining protocol combined with image analysis is accurate and less labor-intensive than classical procedures for determining the cross-sectional area of myofibers.

Mozdziak, P. E.; Fassel, T. A.; Schultz, E.; Greaser, M. L.; Cassens, R. G.

1996-01-01

124

Diagnostic value of direct fluorescence antibody staining for detecting Pneumocystis jirovecii in expectorated sputum from patients with HIV infection.  

PubMed

Direct fluorescent antibody (DFA) staining of induced sputum is frequently used to diagnose Pneumocystis pneumonia (PCP) in patients infected with human immunodeficiency virus, although induction can provoke nausea and bronchospasm. Since the diagnostic value of expectorated sputum examined with DFA stain has not been well evaluated, we reviewed the medical records of HIV-infected patients who were clinically diagnosed as having PCP between 1999 and 2011. Over this 13-year period, we found 76 patients whose records included the results of DFA staining of expectorated sputum and noted that 42 (55.3%) were positive. Polymerase chain reaction to detect Pneumocystis in the sputum of 65 of the patients resulted in the finding of 43 (66.2%) who were positive. Our findings suggest that DFA staining of expectorated sputum could be a useful initial diagnostic method in HIV-infected patients with PCP. PMID:24667822

Choe, Pyoeng Gyun; Kang, Yoo Min; Kim, Gayeon; Park, Wan Beom; Park, Sang Won; Kim, Hong Bin; Oh, Myoung-Don; Kim, Eui Chong; Kim, Nam Joong

2014-04-01

125

Collective fluorescence switching of counterion-assembled dyes in polymer nanoparticles  

NASA Astrophysics Data System (ADS)

The current challenge in the field of fluorescent nanoparticles (NPs) for bioimaging is to achieve extreme brightness and external control of their emission using biodegradable materials. Here we propose a new concept of fluorescent polymer NPs, doped with ionic liquid-like salts of a cationic dye (octadecyl rhodamine B) with a bulky hydrophobic counterion (fluorinated tetraphenylborate) that serves as spacer minimizing dye aggregation and self-quenching. The obtained 40-nm poly(D,L-lactide-co-glycolide) NPs containing up to 500 dyes are brighter than quantum dots and exhibit photo-induced reversible on/off fluorescence switching, never reported for dye-doped NPs. We show that this collective switching of hundreds of dyes is due to ultrafast excitation energy transfer and can be used for super-resolution imaging. These NPs, being spontaneously endocytosed by living cells, feature high signal-to-noise ratio and absence of toxicity. The counterion-based concept opens the way to a new class of nanomaterials for sensing, imaging and light harvesting.

Reisch, Andreas; Didier, Pascal; Richert, Ludovic; Oncul, Sule; Arntz, Youri; Mély, Yves; Klymchenko, Andrey S.

2014-06-01

126

Comparative studies of pollen and fluorescent dye transport by bumble bees visiting Erythronium grandiflorum  

Microsoft Academic Search

In the Colorado Rocky Mountains the glacier lily Erythronium grandiflorum exhibits a striking dimorphism in pollen color and is commonly pollinated by the bumble bee Bombus occidentalis. We induced bees to visit sequences of flowers in a flight cage, and compared dispersal of distinctively-colored pollen and fluorescent pigment (“dye”) that the bee had picked up at a single donor flower.

James D. Thomson; Mary V. Price; Nickolas M. Waser; Donald A. Stratton

1986-01-01

127

Use of Tunable, Pulsed Dye Laser for Quantitative Fluorescence in Syphilis Serology (FTA-ABS Test)  

PubMed Central

A pulsed dye laser was used as an excitation source in a fluorescent treponemal antibody absorption (FTA-ABS) test. A high precision in quantitative fluorescence was obtained with this high-power excitation source coupled to an electronic detection system and a storage oscilloscope by standardization of fluorescence evaluation and through elimination of human error. One 0.4-?s pulse exposure was sufficient to record fluorescence intensity data on the oscilloscope. Absence of fading of fluorescence after repeated excitation permitted multiple readings of the same microscope field. Almost 100% reproducible results were obtained for the FTA-ABS test with 40 samples. Electronic detection of fluorescence and the high sensitivity obtained with laser excitation raise doubts about the relative value of quantitative immunofluorescence in the FTA-ABS test. PMID:4598221

Kasatiya, S. S.; Lambert, N. G.; Laurence, R. A.

1974-01-01

128

Acid-fast stain  

MedlinePLUS

The acid-fast stain is a laboratory test that determines if a sample of tissue, blood, or other body ... dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria ...

129

Miniaturized side-viewing imaging probe for fluorescence lifetime imaging (FLIM): validation with fluorescence dyes, tissue structural proteins and tissue specimens  

Microsoft Academic Search

We report a side viewing fibre-based endoscope that is compatible with intravascular imaging and fluorescence lifetime imaging microscopy (FLIM). The instrument has been validated through testing with fluorescent dyes and collagen and elastin powders using the Laguerre expansion deconvolution technique to calculate the fluorescence lifetimes. The instrument has also been tested on freshly excised unstained animal vascular tissues.

D S Elson; J A Jo; L Marcu

2007-01-01

130

Design and synthesis of a novel fluorescent protein probe for easy and rapid electrophoretic gel staining by using a commonly available UV-based fluorescent imaging system.  

PubMed

A new fluorescent molecular probe, methyl 3-(3,5-bis((bis(pyridin-2-ylmethyl)amino)-methyl)-4-hydroxyphenyl)-2-(5-(dimethylamino)naphthalene-1-sulfonamido) propanoate, dizinc(II) chloride salt (Dansyl-1-Zn(II)), which possesses Zn(II) complexes and a dansyl group, was designed and synthesized to enable the detection of proteins in solution and in high-throughput electrophoresis by using a UV-based detection system. Dansyl-1-Zn(II) exhibited weak fluorescence in the absence of proteins and strong green fluorescence at approximately 510 nm in the presence of BSA upon irradiation with light at a wavelength of 345 nm. Compared with conventional protocols for in-gel SDS-PAGE protein staining (e.g. silver staining, SYPRO Ruby, and Oriole), the operating times of which range from 90 min to overnight, Dansyl-1-Zn(II) allowed 1-step protein staining (SDS-PAGE ?Staining ?Detection) and shortened the operating time (35 min) with high sensitivity (LOD: 1 ng or less) under 312-nm or 365-nm light excitation with orange or red emission filters, respectively. Moreover, Dansyl-1-Zn(II) was successfully applied to protein identification by MS via in-gel tryptic digestion, Western blotting, and Native-PAGE. Accordingly, Dansyl-1-Zn(II) may facilitate highly sensitive and high-throughput protein detection, and it may be widely applicable as a convenient tool in various scientific and medical fields. PMID:23801451

Suzuki, Yoshio; Takagi, Nobuyuki; Sano, Takuma; Chimuro, Tomoyuki

2013-09-01

131

Synchronous fluorescence spectroscopic study of solvatochromic curcumin dye  

NASA Astrophysics Data System (ADS)

Curcumin, the main yellow bioactive component of turmeric, has recently acquired attention by chemists due its wide range of potential biological applications as an antioxidant, an anti-inflammatory, and an anti-carcinogenic agent. This molecule fluoresces weakly and poorly soluble in water. In this detailed study of curcumin in thirteen different solvents, both the absorption and fluorescence spectra of curcumin was found to be broad, however, a narrower and simple synchronous fluorescence spectrum of curcumin was obtained at ? ? = 10-20 nm. Lippert-Mataga plot of curcumin in different solvents illustrated two sets of linearity which is consistent with the plot of Stokes' shift vs. the ET30. When Stokes's shift in wavenumber scale was replaced by synchronous fluorescence maximum in nanometer scale, the solvent polarity dependency measured by ?SFSmax vs. Lippert-Mataga plot or ET30 values offered similar trends as measured via Stokes' shift for protic and aprotic solvents for curcumin. Better linear correlation of ?SFSmax vs. ?* scale of solvent polarity was found compared to ?absmax or ?emmax or Stokes' shift measurements. In Stokes' shift measurement both absorption/excitation as well as emission (fluorescence) spectra are required to compute the Stokes' shift in wavenumber scale, but measurement could be done in a very fast and simple way by taking a single scan of SFS avoiding calculation and obtain information about polarity of the solvent. Curcumin decay properties in all the solvents could be fitted well to a double-exponential decay function.

Patra, Digambara; Barakat, Christelle

2011-09-01

132

Development of a quantitative structure activity relations (QSAR) model to guide the design of fluorescent dyes for detecting amyloid fibrils.  

PubMed

Quantitative structure activity relationship (QSAR) studies were performed on a set of polymethine compounds to develop new fluorescent probes for detecting amyloid fibrils. Two different approaches were evaluated for developing a predictive model: part least squares (PLS) regression and an artificial neural network (ANN). A set of 60 relevant molecular descriptors were selected by performing principal component analysis on more than 1600 calculated molecular descriptors. Through QSAR analysis, two predictive models were developed. The final versions produced an average prediction accuracy of 72.5 and 84.2% for the linear PLS and the non-linear ANN procedures, respectively. A test of the ANN model was performed by using it to predict the activity, i.e., staining or non-staining of amyloid fibrils, using 320 compounds. The five candidates whose greatest activities were selected by the ANN model underwent confirmation of their predicted properties by empirical testing. The results indicated that the ANN model potentially is useful for facilitating prediction of activity of untested compounds as dyes for detecting amyloid fibrils. PMID:24251531

Inshyn, D I; Kovalska, V B; Losytskyy, M Y; Slominskii, Yl; Tolmachev, O I; Yarmoluk, S M

2014-01-01

133

High resolution deformation and damage detection using fluorescent dyes  

NASA Astrophysics Data System (ADS)

We demonstrate the application of fluorescence microscopy to detect nanoscale deformation and damage in polymeric materials. Fluorescent probes were dispersed in a poly-dimethyl siloxane matrix, and were subsequently strained with and without the presence of edge cracks. This technique can reveal cracks that are invisible to white light microscopy (smaller than 250 nm), and is outperformed only by high resolution electron or scanning probe microscopy. The technique may find applications in early stage damage detection in structural health monitoring systems for a wide variety of polymeric materials.

Samuel, Benedict A.; Demirel, Melik C.; Haque, Aman

2007-11-01

134

Polymerase chain reaction for verification of fluorescent colonies of Erwinia chrysanthemi and Pseudomonas putida WCS358 in immunofluorescence colony staining.  

PubMed

The potential of polymerase chain reaction (PCR) for verifying the identity of colonies stained by the immunofluorescence colony-staining (IFC) procedure was investigated. Using primers directed against conserved sequences of the pectate lyase-genes coding for isozymes PLa, PLd and PLe of Erwinia chrysanthemi, the authors confirmed the identity of 96% of 20 fluorescent target colonies, punched from IFC-stained samples with pure cultures. In pour plates with mixtures of Erw. chrysanthemi and non-target colonies from potato peel extracts, the identity of 90% of 113 target colonies was confirmed. Using primers directed against sequences of the ferric-pseudobactin receptor gene pupA of Pseudomonas putida WCS358, the identity of 96% of 22 target colonies was confirmed in IFC-stained samples with pure cultures. In pour plates with mixtures of Ps. putida WCS358 and non-target bacteria from compost extracts, the identity of 59% of 108 fluorescent colonies was confirmed by PCR. It was shown that components from non-target bacteria lowered the threshold level of PCR for Ps. putida WCS358 100-fold. PMID:8567494

van der Wolf, J M; van Beckhoven, J R; de Vries, P M; Raaijmakers, J M; Bakker, P A; Bertheau, Y; van Vuurde, J W

1995-11-01

135

Sentinel lymph nodes detection with an imaging system using Patent Blue V dye as fluorescent tracer  

NASA Astrophysics Data System (ADS)

Sentinel lymph node biopsy is the gold standard to detect metastatic invasion from primary breast cancer. This method can help patients avoid full axillary chain dissection, thereby decreasing the risk of morbidity. We propose an alternative to the traditional isotopic method, to detect and map the sentinel lymph nodes. Indeed, Patent Blue V is the most widely used dye in clinical routine for the visual detection of sentinel lymph nodes. A Recent study has shown the possibility of increasing the fluorescence quantum yield of Patent Blue V, when it is bound to human serum albumin. In this study we present a preclinical fluorescence imaging system to detect sentinel lymph nodes labeled with this fluorescent tracer. The setup is composed of a black and white CCD camera and two laser sources. One excitation source with a laser emitting at 635 nm and a second laser at 785 nm to illuminate the region of interest. The prototype is operated via a laptop. Preliminary experiments permitted to determine the device sensitivity in the ?mol.L-1 range as regards the detection of PBV fluorescence signals. We also present a preclinical evaluation performed on Lewis rats, during which the fluorescence imaging setup detected the accumulation and fixation of the fluorescent dye on different nodes through the skin.

Tellier, F.; Steibel, J.; Chabrier, R.; Rodier, J. F.; Pourroy, G.; Poulet, P.

2013-03-01

136

Measurement of atmospheric OH by titration of near-IR fluorescent dyes  

NASA Technical Reports Server (NTRS)

Recent research has shown that certain polymethine dyes can be detected at ultratrace levels (greater than or equal to 6x10(exp -14) M) in solution by fluorimetry. These detection limits are possible because of the inherent sensitivity of fluorescence techniques, because the dyes fluoresce in the near infrared region where background interference is negligible, and because powerful infrared diode lasers are now available to improve the signal to noise ratio. Other work has shown that the hydroxyl radical destroys the ability of polymethine dyes to fluoresce. These observations form the basis for a new hydroxyl radical detector that is essentially a fluorometric titrator. Theoretically, the detector should show an acceptable sensitivity and response time. Assuming that the atmospheric HO concentration is about 10(exp -11) moles m(exp -3) (i.e. 10(exp 6) molecules cm(exp -3)), then 10 L of air 'titrated' with 20 mL of 10(exp -11) M dye solution (an easily detected concentration) should result in a drop in the fluorescent signal of 50 percent - a readily detectable change. At a flow rate of 3 L min(exp -1) the sampling time would be 3 minutes. The biggest potential problem is selectivity: other oxidants may also cause the fluorescence signal to be lost. The chemistry of polymethine dyes has not been studied in detail and so no quantitative data are available. However, a survey of the literature suggests that in general HO should react up to six orders of magnitude faster than HO2 and other radicals such as RO2 and RO. It should also react much more rapidly than H2O2 and O3. Thus it may be possible to discriminate kinetically against potential interfering substances. It was shown in the laboratory that 10(exp -4) M H2O2 has little effect on the absorption spectrum of the dye IR125 over a period of hours but that the band at 780 nm is slowly lost in water over a period of days even under argon in the dark. By contrast, DMSO solutions of IR125 are stable.

Betterton, Eric A.; Gast, Karl

1994-01-01

137

Fluorescence quenching and photocatalytic degradation of textile dyeing waste water by silver nanoparticles  

NASA Astrophysics Data System (ADS)

Silver nanoparticles (Ag NPs) of different sizes have been prepared by chemical reduction method and characterized using UV-vis spectroscopy and transmission electron microscopy (HRTEM). Fluorescence spectral analysis showed that the quenching of fluorescence of textile dyeing waste water (TDW) has been found to decrease with decrease in the size of the Ag NPs. Experimental results show that the silver nanoparticles can quench the fluorescence emission of adsorbed TDW effectively. The fluorescence interaction between Ag NPs (acceptor) and TDW (donor) confirms the Förster Resonance Energy Transfer (FRET) mechanism. Long range dipole-dipole interaction between the excited donor and ground state acceptor molecules is the dominant mechanism responsible for the energy transfer. Furthermore, photocatalytic degradation of TDW was measured spectrophotometrically by using silver as nanocatalyst under UV light illumination. The kinetic study revealed that synthesized Ag NPs was found to be effective in degrading TDW.

Kavitha, S. R.; Umadevi, M.; Janani, S. R.; Balakrishnan, T.; Ramanibai, R.

2014-06-01

138

Oligonucleotide assisted light-emitting Alq3 microrods: energy transfer effect with fluorescent dyes.  

PubMed

Oligonucleotide assisted tri(8-hydroxyquinoline) aluminium (Alq3) microrods were prepared for the first time. When hybridized with oligonucleotide labeled by Cy3 fluorescent dye, a significant photoluminescence variation of the Alq3 microrods was observed due to Förster resonance energy transfer, unlike when Cy5-oligonucleotide was used. Versatile nucleotide manipulation would open up wider applications of Alq3-based materials, based on this fundamental observation. PMID:23535632

Cui, Chunzhi; Park, Dong Hyuk; Kim, Jeongyong; Joo, Jinsoo; Ahn, Dong June

2013-06-14

139

Study of excitation transfer in laser dye mixtures by direct measurement of fluorescence lifetime  

NASA Technical Reports Server (NTRS)

By directly measuring the donor fluorescence lifetime as a function of acceptor concentration in the laser dye mixture Rhodamine 6G-Cresyl violet, we found that the Stern-Volmer relation is obeyed, from which the rate of excitation transfer is determined. The experimental results indicate that the dominant mechanism responsible for the efficient excitation transfer is that of resonance transfer due to long range dipole-dipole interaction.

Lin, C.; Dienes, A.

1973-01-01

140

Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.  

PubMed Central

Fluorescent dye-labeled DNA primers have been developed that exploit fluorescence energy transfer (ET) to optimize the absorption and emission properties of the label. These primers carry a fluorescein derivative at the 5' end as a common donor and other fluorescein and rhodamine derivatives attached to a modified thymidine residue within the primer sequence as acceptors. Adjustment of the donor-acceptor spacing through the placement of the modified thymidine in the primer sequence allowed generation of four primers, all having strong absorption at a common excitation wavelength (488 nm) and fluorescence emission maxima of 525, 555, 580, and 605 nm. The ET efficiency of these primers ranges from 65% to 97%, and they exhibit similar electrophoretic mobilities by gel electrophoresis. With argon-ion laser excitation, the fluorescence of the ET primers and of the DNA sequencing fragments generated with ET primers is 2- to 6-fold greater than that of the corresponding primers or fragments labeled with single dyes. The higher fluorescence intensity of the ET primers allows DNA sequencing with one-fourth of the DNA template typically required when using T7 DNA polymerase. With single-stranded M13mp18 DNA as the template, a typical sequencing reaction with ET primers on a commercial sequencer provided DNA sequences with 99.8% accuracy in the first 500 bases. ET primers should be generally useful in the development of other multiplex DNA sequencing and analysis methods. Images Fig. 4 Fig. 5 PMID:7753809

Ju, J; Ruan, C; Fuller, C W; Glazer, A N; Mathies, R A

1995-01-01

141

Determination of blood plasma fluorescence extinction coefficients for dyes used in three-compartment binding model  

NASA Astrophysics Data System (ADS)

A three-compartment kinetic model for the binding of a ligand to its receptor in tumor tissue has been explained and the kinetic rates of the model are currently being investigated. In order to determine the plasma excretion rates of the dyes of interest, the fluorescence extinction coefficients must be determined. The fluorescence extinction coefficients of the IRDye700DX-carboxylate (IRDye700DX-C) and IRDye800CW-conjugated to EGFR (IRDye800CW-EGF) have been to be 7.98 ×106 ?M-1 cm-1 and 4.73x106 ?M-1 cm-1, respectively. We determined that the linear range of these dyes in the blood plasma of a mouse was 0 - 0.26 ?M. Administration of 1 nmol of each of these dyes to a mouse weighing 25-30g (0.04 ?M - 0.033 ?M, respectively) will result in blood plasma fluorescence in the linear and readable range.

Samkoe, Kimberley S.; Sexton, Kristian; Tichauer, Kenneth; Davis, Scott C.; O'Hara, Julia A.; Hasan, Tayyaba; Pogue, Brian W.

2011-02-01

142

Fluorescence intensity and anisotropy decays of the DNA stain Hoechst 33342 resulting from one-photon and two-photon excitation  

NASA Astrophysics Data System (ADS)

We studied the steady state and time-resolved fluorescence spectral properties of the DNA stain Hoechst 33342 for one-photon (OPE) and two-photon (TPE) excitation. Hoechst 33342 was found to display a large cross-section for two-photon excitation within the fundamental wavelength range of pyridine 2 and rhodamine 6G dye lasers, 690 to 770 nm and 560 to 630 nm, respectively. The time-resolved measurements show that intensity decays are similar for one- and two-photon excitation. The anisotropy decay measurements of bis-benzimide, 2,5'-bi-1H-benzimidazole, 2'-(4- ethoxphenyl)-5-(4-methyl-1-piperazinyl) (HOECHST 33342) in ethanol revealed the same correlation times for two-photon excitation as observed for one-photon excitation. However, the zero-time anisotropies recovered from anisotropy decay measurements are 1.4-fold higher for two-photon excitation than for one-photon excitation. The anisotropy spectra of Hoechst 33342 was examined in glycerol at -20 degree(s)C, revealing limiting values close to the theoretical limits for one-photon (0.4) and two-photon (0.57) excitation. The steady-state anisotropy for one-photon excitation decreases in the shorter wavelength region (R6G dye laser, 280 to 315 nm), but the two-photon anisotropy for 560 to 630 nm excitation remains as high as in the long- wavelength region (690 to 770 nm). This result suggests that one- photon absorption is due to two electronic transitions, but only one transition contributes to the two-photon absorption over the wavelength range from 580 to 770 nm.

Gryczynski, Ignacy; Lakowicz, Joseph R.

1994-08-01

143

Conjugates of a Photoactivated Rhodamine with Biopolymers for Cell Staining  

PubMed Central

Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids) are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan (“Chitosan-PFD”) and histone H1 (“Histone H1.3-PFD”). The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK). Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes (“caged” dyes) for microscopic probing of biological objects. Thus, the synthesized “Chitosan-PFD” and “Histone H1-PFD” have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy. PMID:25383365

Zaitsev, Sergei Yu.; Shaposhnikov, Mikhail N.; Solovyeva, Daria O.; Solovyeva, Valeria V.; Rizvanov, Albert A.

2014-01-01

144

Dictyostelium Extracellular Vesicles Containing Hoechst 33342 Transfer the Dye into the Nuclei of Living Cells: A Fluorescence Study  

Microsoft Academic Search

Cells of the eukaryotic unicellular microorganism Dictyostelium discoideum are constitutively resistant to vital staining of their nuclei by the DNA-specific dye Hoechst 33342. By studying the mechanisms\\u000a of this resistance, we evidenced that these cells expel vesicles containing the dye for detoxification (Tatischeff et al.,\\u000a Cell Mol Life Sci, 54: 476–87, 1998). The question to be addressed in the present

Irène Tatischeff; Françoise Lavialle; Sophie Pigaglio-Deshayes; Christine Péchoux-Longin; Laurent Chinsky; Annette Alfsen

2008-01-01

145

Blood analyte sensing using fluorescent dye-loaded red blood cells  

NASA Astrophysics Data System (ADS)

Measurement of blood analytes provides crucial information about a patient's health. Some such analytes, such as glucose in the case of diabetes, require long-term or near-continuous monitoring for proper disease management. However, current monitoring techniques are far from ideal: multiple-per-day finger stick tests are inconvenient and painful for the patient; implantable sensors have short functional life spans (i.e., 3-7 days). Due to analyte transporters on red blood cell (RBC) membranes that equilibrate intracellular and extracellular analyte levels, RBCs serve as an attractive alternative for encapsulating analyte sensors. Once reintroduced to the blood stream, the functionalized RBCs may continue to live for the remainder of their life span (120 days for humans). They are biodegradable and biocompatible, thereby eliminating the immune system response common for many implanted devices. The proposed sensing system utilizes the ability of the RBCs to swell in response to a decrease in the osmolarity of the extracellular solution. Just before lysis, they develop small pores on the scale of tens of nanometers. While at low temperature, analyte-sensitive dyes in the extracellular solution diffuse into the perforated RBCs and become entrapped upon restoration of temperature and osmolarity. Since the fluorescent signal from the entrapped dye reports on changes in the analyte level of the extracellular solution via the RBC transporters, interactions between the RBCs and the dye are critical to the efficacy of this technique. In this work, we study the use of a near infrared pH sensitive dye encapsulated within RBCs and assess the ability to measure dye fluorescence in vivo.

Ritter, Sarah C.; Shao, Xiaole; Cooley, Nicholas; Milanick, Mark A.; Glass, Timothy E.; Meissner, Kenith E.

2014-02-01

146

Confocal laser scanning microscopy of mitochondria within microspore tetrads of plants using rhodamine 123 as a fluorescent vital stain.  

PubMed

The present study demonstrates that rhodamine 123 penetrates the callose walls surrounding plant microspores before they are released from tetrads. The stain accumulates in active mitochondria due to the electrical potential across the mitochondrial membrane. Accumulation of dye does not occur in mitochondria of fixed cells and fades quickly when mitochondrial activity is inhibited by exposure to carbonyl cyanide m-chlorophenyl hydrazone. Rhodamine can be used as a viability test for microspores still within tetrads, thus making it possible to determine when during development genes leading to pollen sterility are expressed. Rhodamine 123 is excited by blue (550 nm) light and can thus be used with confocal laser scanning microscopy. Anthers of Nicotiana tabacum, Oenothera villaricae, Silene dioica and Lycopersicum esculentum were studied here. PMID:7703302

Gambier, R M; Mulcahy, D L

1994-11-01

147

Detection of acid moisture in photovoltaic modules using a dual wavelength pH-sensitive fluorescent dye  

NASA Astrophysics Data System (ADS)

The formation of acetic acid via the penetration of moisture into ethylene vinyl acetate (EVA) in photovoltaic (PV) modules is cited as the main reason for PV modules’ degradation. Currently, there is no effective method for detecting acetic moisture in PV modules. We proposed a simple method for detecting acid moisture in PV modules using a dual-wavelength pH-sensitive dye that measures pH by the ratio of the intensities of two peaks in the fluorescence spectra of the dye. We detected the pH change caused by acetic acid with the change in the intensity ratio of the fluorescence spectra of the dried dye. Furthermore, we observed that the dry fluorescent dye is heat resistant to withstand the lamination process for the manufacturing of PV modules, and has good long-term durability.

Asaka, Takashi; Iwami, Kentaro; Taguchi, Atsushi; Umeda, Norihiro; Masuda, Atsushi

2014-01-01

148

Reduction of Nonspecific Background Staining in the Fluorescent Treponemal Antibody-Absorption Test  

PubMed Central

The nonspecific background fluorescence which occurs with the fluorescent treponemal antibody-absorption test for syphilis was found to result from a reaction between serum-treated Treponema pallidum organisms and the conjugated antihuman ?-globulin. It was also shown that ?-lipoprotein and albumin were the important contributing factors in human serum. Various dilutions of 2.5% trypsin in phosphate-buffered saline specifically reduced background fluorescence under proper test conditions. By employing a trypsin digestion method, a semiautomated procedure utilizing a visual readout has been postulated as feasible. PMID:4177869

Roberts, Merritt E.; Miller, James N.; Binnings, Gerald F.

1968-01-01

149

Staining paraffin extracted, alcohol rinsed and air dried plant tissue with an aqueous mixture of three dyes.  

PubMed

A staining solution containing alcian blue 8GX, Bismarck brown Y and safranin O was prepared with 0.1 M sodium acetate buffer, pH 5.0. Paraffin was extracted with MicroClear solvent from 10 microm tissue sections mounted on slides. Paraffin solvent was removed by rinsing with isopropanol, and tissues were air dried. Slides with bare dry tissue sections were immersed in the triple stain and structures could be distinguished within 30 min as follows: nonlignified cell walls, blue; lignified cell walls, nuclei and chloroplasts, red; and cuticle, brown or yellow-brown. Excess staining solution was removed by rinsing with tap water, and the tissues were air dried again. Coverslips were affixed with resin over the stained dry tissues. This novel procedure was tested with immature tomato fruit, mature apple fruit, and various leaf and stem specimens of dogwood, laurel, pawpaw, poinsettia and zonal geranium. PMID:9735876

Graham, E T; Trentham, W R

1998-07-01

150

Time-resolved fluorescence lifetime imaging microscopy using a picosecond pulsed tunable dye laser system  

NASA Astrophysics Data System (ADS)

The design and implementation of a time-resolved fluorescence lifetime imaging microscope (TRFLIM) for the biomedical sciences are described. The measurement of fluorescence lifetimes offers many benefits, among which is that they are independent of local signal intensity and concentration of the fluorophore and they provide visualization of the molecular environment in a single living cell. Unlike single photon counting, which employs a photomultiplier as the detector, TRFLIM uses a nanosecond-gated multichannel plate image intensifier providing a two-dimensional map of the spatial distribution of fluorescent lifetime in the sample under observation. Picosecond laser pulses from a tunable dye laser are delivered to the fluorophore inside living cells on the stage of a fluorescent microscope. Images of the fluorescence emission at various times during the decay of the fluorescence are collected using a high-speed gated image intensifier and the lifetimes are calculated on a pixel-by-pixel basis. Lifetimes measured by TRFLIM are compared with those measured by conventional methods.

Periasamy, Ammasi; Wodnicki, Pawel; Wang, Xue F.; Kwon, Seongwook; Gordon, Gerald W.; Herman, Brian

1996-10-01

151

Is the central dogma of flow cytometry true: that fluorescence intensity is proportional to cellular dye content  

SciTech Connect

Measurements and theoretical calculations of fluorescent emission from four samples of polystyrene microspheres (diameter 0.92, 1.63, 1.90 and 4.18 ..mu..m) containing the same fluorescent dye show a general dependence upon particle size, emission angle, and polarization conditions. However, for the excitation and detection conditions used in flow cytometry, the relative fluorescent intensities measured for the four particle sizes are proportional to the dye content to +10% accuracy, independent of particle size. Accordingly, the central dogma of flow cytometry that fluorescence is proportional to cellular dye content is valid to this accuracy for these solid, highly refractive polymer particles. Most mammalian cells are much less refractive, therefore, should conform more closely to the central dogma.

Kerker, M.; Van Dilla, M.A.; Brunsting, A.; Kratohvil, J.P.; Hsu, P.; Wang, D.S.; Gray, J.W.; Langlois, R.G.

1982-01-01

152

Fluorescent Sensing of Chlorophenols in Water Using an Azo Dye Modified ?-Cyclodextrin Polymer  

PubMed Central

A water soluble azo dye modified ?-cyclodextrin polymer 4 was synthesized and used as a chemosensor for the detection of chlorinated phenols, model chlorinated by-products (CBPs) of water treatment for drinking purposes. The characterization of the intermediates and the azo dye modified ?-CD polymer was done by UV/Vis Spectrophotometry, FT-IR and 1H-NMR spectroscopies. The chlorophenols were capable of quenching the fluorescence of the polymer. The polymer showed greater sensitivity towards 2,4-dichlorophenol, with a sensitivity factor of 0.35 compared to 0.05 and 0.12 for phenol and 4-chlorophenol, respectively. The stability constants (Ks) of the pollutants were also determined by the Benesi-Hildebrand method to be 2.104 × 103 M?1 for 2,4-dichlorophenol and 1.120 × 102 M?1 for 4-chlorophenol. PMID:22163864

Ncube, Phendukani; Krause, Rui W.; Mamba, Bhekie B.

2011-01-01

153

Is the central dogma of flow cytometry true: that fluorescence intensity is proportional to cellular dye content  

Microsoft Academic Search

Measurements and theoretical calculations of fluorescent emission from four samples of polystyrene microspheres (diameter 0.92, 1.63, 1.90 and 4.18 ..mu..m) containing the same fluorescent dye show a general dependence upon particle size, emission angle, and polarization conditions. However, for the excitation and detection conditions used in flow cytometry, the relative fluorescent intensities measured for the four particle sizes are proportional

M. Kerker; M. A. Van Dilla; A. Brunsting; J. P. Kratohvil; P. Hsu; D. S. Wang; J. W. Gray; R. G. Langlois

1982-01-01

154

Quantitative diagnosis of cervical neoplasia using fluorescence lifetime imaging on haematoxylin and eosin stained tissue sections.  

PubMed

The use of conventional fluorescence microscopy for characterizing tissue pathological states is limited by overlapping spectra and the dependence on excitation power and fluorophore concentration. Fluorescence lifetime imaging microscopy (FLIM) can overcome these limitations due to its insensitivity to fluorophore concentration, excitation power and spectral similarity. This study investigates the diagnosis of early cervical cancer using FLIM and a neural network extreme learning machine classifier. A concurrently high sensitivity and specificity of 92.8% and 80.2%, respectively, were achieved. The results suggest that the proposed technique can be used to supplement the traditional histopathological examination of early cervical cancer. PMID:23281280

Gu, Jun; Fu, Chit Yaw; Ng, Beng Koon; Gulam Razul, Sirajudeen so; Lim, Soo Kim

2014-07-01

155

Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes.  

PubMed

To enable studies of conformational changes within multimolecular complexes, we present a simultaneous, four-color single molecule fluorescence methodology implemented with total internal reflection illumination and camera-based, wide-field detection. We further demonstrate labeling histidine-tagged proteins noncovalently with Tris-nitrilotriacetic acid (Tris-NTA)-conjugated dyes to achieve single molecule detection. We combine these methods to colocalize the mismatch repair protein MutS? on DNA while monitoring MutS?-induced DNA bending using Förster resonance energy transfer (FRET) and to monitor assembly of membrane-tethered SNARE protein complexes. PMID:21091445

DeRocco, Vanessa; Anderson, Trevor; Piehler, Jacob; Erie, Dorothy A; Weninger, Keith

2010-11-01

156

Application of Temperature-Dependent Fluorescent Dyes to the Measurement of Millimeter Wave Absorption in Water Applied to Biomedical Experiments  

PubMed Central

Temperature sensitivity of the fluorescence intensity of the organic dyes solutions was used for noncontact measurement of the electromagnetic millimeter wave absorption in water. By using two different dyes with opposite temperature effects, local temperature increase in the capillary that is placed inside a rectangular waveguide in which millimeter waves propagate was defined. The application of this noncontact temperature sensing is a simple and novel method to detect temperature change in small biological objects. PMID:25435859

Popenko, Oleksandr

2014-01-01

157

Application of temperature-dependent fluorescent dyes to the measurement of millimeter wave absorption in water applied to biomedical experiments.  

PubMed

Temperature sensitivity of the fluorescence intensity of the organic dyes solutions was used for noncontact measurement of the electromagnetic millimeter wave absorption in water. By using two different dyes with opposite temperature effects, local temperature increase in the capillary that is placed inside a rectangular waveguide in which millimeter waves propagate was defined. The application of this noncontact temperature sensing is a simple and novel method to detect temperature change in small biological objects. PMID:25435859

Kuzkova, Nataliia; Popenko, Oleksandr; Yakunov, Andrey

2014-01-01

158

Gram stain  

MedlinePLUS

A Gram stain is a test used to identify bacteria. It is one of the most common ways to ... Urethral discharge Gram stain; Feces Gram stain; Stool Gram stain; Joint fluid Gram stain; Pericardial fluid Gram stain; Gram stain of ...

159

On the incorporation of Rhodamine B and 2?,7?-dichlorofluorescein dyes in silica: Synthesis of fluorescent nanoparticles  

NASA Astrophysics Data System (ADS)

The present paper reports the incorporation of 2?,7?-dichlorofluorescein (DCF) and Rhodamine B (RhB) dyes in silica nanoparticles by using the Stöber's method with some modifications. Based on infrared and electronic spectroscopies, these dyes were successfully incorporated resulting in fluorescent nanomaterials of an average size of 80 nm. A composite fluorescent nanomaterial containing both dyes was also synthesized and showed the occurrence of Förster resonant energy transfer process (FRET) with the average distance between the donor (DCF) and acceptor (RhB) of 3.6 nm. Furthermore, these fluorescent nanoparticles were modified with folic acid producing nanomaterials whose Zeta potential values were in the range of -2 to -13 mV. These values are consistent with the low dispersivity observed by TEM micrographs. Altogether, these suitable properties can lead to the development of nanomaterials for cancer bioimaging and drug release.

Gomes, Elis C. C.; de Carvalho, Idalina M. M.; Diógenes, Izaura C. N.; de Sousa, Eduardo H. S.; Longhinotti, Elisane

2014-05-01

160

Micelles assembled with carbocyanine dyes for theranostic near-infrared fluorescent cancer imaging and photothermal therapy.  

PubMed

It is an emerging focus to explore a theranostic nanocarrier for simultaneous cancer imaging and therapy. Herein, we demonstrate a theranostic micelle system for cancer near infrared fluorescent (NIRF) imaging with enhanced signal to noise ratio and superior photothermal therapy. The copolymers consisting of monomethoxy poly(ethylene glycol) and alkylamine-grafted poly(L-aspartic acid) are assembled with carbocyanine dyes into theranostic micelles, which exhibit small size, high loading capacity, good stability, sustained release behavior, and enhanced cellular uptake. The micelles achieve the preferable biodistribution and long-term retention of carbocyanine dyes at tumor, which result in enhanced NIRF imaging by generating stable retention of NIRF signals at both hypervascular and hypovascular tumors during a long-term imaging period of up to 8 day, accompanying with negligible noise at normal tissues. The photostability of carbocyanine dye (Cypate) plays an important role for long-term cancer imaging with enhanced SNR. Moreover, the micelles exhibit severe photothermal damage on cancer cells via the destabilization of subcellular organelles upon photoirradiation, causing superior photothermal tumor regress. The micelles act as a powerful theranostic nanocarrier for simultaneous cancer imaging with high contrast and superior photothermal therapy. PMID:24008037

Yang, Hong; Mao, Huajian; Wan, Zhihui; Zhu, Aijun; Guo, Miao; Li, Yanli; Li, Xinming; Wan, Jiangling; Yang, Xiangliang; Shuai, Xintao; Chen, Huabing

2013-12-01

161

Detection of microlesions induced by heavy ions using liposomes filled with fluorescent dye  

NASA Technical Reports Server (NTRS)

In cells irradiation by heavy ions has been hypothesized to produce microlesions, regions of local damage. In cell membranes this damage is thought to manifest itself in the form of holes. The primary evidence for microlesions comes from morphological studies of cell membranes, but this evidence is still controversial, especially since holes also have been observed in membranes of normal, nonirradiated, cells. However, it is possible that damage not associated with histologically discernable disruptions may still occur. In order to resolve this issue, we developed a system for detecting microlesions based on liposomes filled with fluorescent dye. We hypothesized that if microlesions form in these liposomes as the result of irradiation, then the entrapped dye will leak out into the surrounding medium in a measurable way. Polypropylene vials containing suspensions of vesicles composed of either dipalmitoyl phosphatidylcholine, or a combination of egg phosphatidylcholine and cholesterol were irradiated at the Brookhaven National Laboratory using 56Fe ions at 1 GeV/amu. In several cases we obtained a significant loss of the entrapped dye above the background level. Our results suggest that holes may form in liposomes as the result of heavy ion irradiation, and that these holes are large enough to allow leakage of cell internal contents that are at least as large as a 1 nm diameter calcein molecule. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

Koniarek, J. P.; Thomas, J. L.; Vazquez, M.

2004-01-01

162

Energy transfer processes in dye-doped nanostructures yield cooperative and versatile fluorescent probes  

NASA Astrophysics Data System (ADS)

Fast and efficient energy transfer among dyes confined in nanocontainers provides the basis of outstanding functionalities in new-generation luminescent probes. This feature article provides an overview of recent research achievements on luminescent Pluronic-Silica NanoParticles (PluS NPs), a class of extremely monodisperse core-shell nanoparticles whose design can be easily tuned to match specific needs for diverse applications based on luminescence, and that have already been successfully tested in in vivo imaging. An outline of their outstanding properties, such as tuneability, bright and photoswitchable fluorescence and electrochemiluminescence, will be supported by a critical discussion of our recent works in this field. Furthermore, novel data and simulations will be presented to (i) thoroughly examine common issues arising from the inclusion of multiple dyes in a small silica core, and (ii) show the emergence of a cooperative behaviour among embedded dyes. Such cooperative behaviour provides a handle for fine control of brightness, emission colour and self-quenching phenomena in PluS NPs, leading to significantly enhanced signal to noise ratios.

Genovese, Damiano; Rampazzo, Enrico; Bonacchi, Sara; Montalti, Marco; Zaccheroni, Nelsi; Prodi, Luca

2014-02-01

163

Dye analysis of Shosoin textiles using excitation-emission matrix fluorescence and ultraviolet-visible reflectance spectroscopic techniques.  

PubMed

The dyes of 8th century textiles, treasured for more than 1250 years in the Shosoin treasure repository in Japan, were analyzed by nondestructive methods, i.e., excitation-emission matrix (EEM) fluorescence and ultraviolet-visible (UV-vis) reflectance spectrometry, in combination with natural dye references extracted from plants, which have been widely used from ancient times. In this analysis, five dyes were found in the following objects: embroidered shoes dedicated to Great Buddha of the Todaiji temple by the empress of that time, the cloth lining for a case holding a mirror belonging to the emperor of that time, two rolls of yellow and light green plain-weave silks, and a sleeveless coat used for a musical in a Buddhist ceremony in 752 A.D. EEM fluorescence spectrometry distinguished kihada yellow (Amur cork tree), kariyasu yellow (eulalia), and akane red (Japanese madder). UV-vis spectrometry also distinguished kariyasu yellow, ai blue (knotweed), akane red, and shikon purple (murasaki); the characteristic peaks of these dyes were detected by a second derivatization. The results show that although the dyes used easily degrade with age, EEM fluorescence and UV-vis reflectance spectrometry are useful for distinguishing dyes used in the Shosoin textiles, which had been stored for more than 1250 years. PMID:19507884

Nakamura, Rikiya; Tanaka, Yoko; Ogata, Atsuhiko; Naruse, Masakazu

2009-07-15

164

The study of polyplex formation and stability by time-resolved fluorescence spectroscopy of SYBR Green I-stained DNA.  

PubMed

Polyplexes are nanoparticles formed by the self-assembly of DNA/RNA and cationic polymers specifically designed to deliver exogenous genetic material to cells by a process called transfection. There is a general consensus that a subtle balance between sufficient extracellular protection and intracellular release of nucleic acids is a key factor for successful gene delivery. Therefore, there is a strong need to develop suitable tools and techniques for enabling the monitoring of the stability of polyplexes in the biological environment they face during transfection. In this work we propose time-resolved fluorescence spectroscopy in combination with SYBR Green I-DNA dye as a reliable tool for the in-depth characterization of the DNA/vector complexation state. As a proof of concept, we provide essential information on the assembly and disassembly of complexes formed between DNA and each of three cationic polymers, namely a novel promising chitosan-graft-branched polyethylenimine copolymer (Chi-g-bPEI), one of its building block 2 kDa bPEI and the gold standard transfectant 25 kDa bPEI. Our results highlight the higher information content provided by the time-resolved studies of SYBR Green I/DNA, as compared to conventional steady state measurements of ethidium bromide/DNA that enabled us to draw relationships among fluorescence lifetime, polyplex structural changes and transfection efficiency. PMID:25308511

D'Andrea, Cosimo; Pezzoli, Daniele; Malloggi, Chiara; Candeo, Alessia; Capelli, Giulio; Bassi, Andrea; Volonterio, Alessandro; Taroni, Paola; Candiani, Gabriele

2014-12-01

165

Comparison of Pollen Transfer Dynamics by Multiple Floral Visitors: Experiments with Pollen and Fluorescent Dye  

PubMed Central

• Background and Aims Most plant species are visited by a diversity of floral visitors. Pollen transfer of the four most common pollinating bee species and one nectar-robbing bee of the distylous plant Gelsemium sempervirens were compared. • Methods Naturally occurring pollen loads carried by the common floral visitor species of G. sempervirens were compared. In addition, dyed pollen donor flowers and sequences of four emasculated recipient flowers in field cages were used to estimate pollen transfer, and the utility of fluorescent dye powder as an analogue for pollen transfer was determined. • Key Results Xylocopa virginica, Osmia lignaria and Habropoda laboriosa carried the most G. sempervirens pollen on their bodies, followed by Bombus bimaculatus and Apis mellifera. However, B. bimaculatus, O. lignaria and H. laboriosa transferred significantly more pollen than A. mellifera. Nectar-robbing X. virginica transferred the least pollen, even when visiting legitimately. Dye particles were strongly correlated with pollen grains on a stigma, and therefore provide a good analogue for pollen in this system. The ratio of pollen?:?dye across stigmas was not affected by bee species or interactions between bee species and floral morphology. However, dye transfer was more sensitive than pollen transfer to differences in floral morphology. • Conclusions The results from this study add to a growing body of literature highlighting that floral visitors vary in pollination effectiveness, and that visitors carrying the most pollen on their bodies may not always be the most efficient at depositing pollen on stigmas. Understanding the magnitude of variability in pollinator quality is one important factor for predicting how different pollinator taxa may influence the evolution of floral traits. PMID:16299005

ADLER, LYNN S.; IRWIN, REBECCA E.

2006-01-01

166

A rapid and noninvasive method to detect dried saliva stains from human skin using fluorescent spectroscopy  

PubMed Central

Objective: Saliva is one of the vital fluids secreted in human beings. Significant amount of saliva is deposited on the skin during biting, sucking or licking, and can act as an important source in forensic evidence. An enzyme, ? amylase, gives a characteristic emission spectrum at 345–355 nm when excited at 282 nm and this can be identified by using fluorescent spectroscopy and can help in forensic identification. This study describes a rapid method to detect dried saliva on the human skin by fluorescent spectroscopy. Materials and Methods: This study included 10 volunteers, who deposited their own saliva on skin of their ventral forearm by licking and water on the contralateral arm as control. This study was carried out at Central Leather Research Institute, Chennai. Study design: Ten volunteers deposited their own saliva on skin of their ventral forearm by licking. A control sample of water was deposited at the contralateral arm. Each sample was excited at 282 nm and emission spectrum was recorded. Results: The emission spectra of 10 swab samples taken from dried saliva were characterized at the primary peak of 345 to 355 nm whereas the emission spectrum of water as a control was recorded at 362 nm. Conclusion: The presence of emission spectrum at 345–355 nm with excitation at 282 nm proves to be a strong indicator of saliva deposited on human skin. PMID:21731273

Nanda, Kanwar Deep Singh; Ranganathan, K; Umadevi, KM; Joshua, Elizabeth

2011-01-01

167

Quantification of AAV Particle Titers by Infrared Fluorescence Scanning of Coomassie-Stained Sodium Dodecyl Sulfate–Polyacrylamide Gels  

PubMed Central

Abstract Adeno-associated virus (AAV)-based vectors have gained increasing attention as gene delivery vehicles in basic and preclinical studies as well as in human gene therapy trials. Especially for the latter two—for both safety and therapeutic efficacy reasons—a detailed characterization of all relevant parameters of the vector preparation is essential. Two important parameters that are routinely used to analyze recombinant AAV vectors are (1) the titer of viral particles containing a (recombinant) viral genome and (2) the purity of the vector preparation, most commonly assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) followed by silver staining. An important, third parameter, the titer of total viral particles, that is, the combined titer of both genome-containing and empty viral capsids, is rarely determined. Here, we describe a simple and inexpensive method that allows the simultaneous assessment of both vector purity and the determination of the total viral particle titer. This method, which was validated by comparison with established methods to determine viral particle titers, is based on the fact that Coomassie Brilliant Blue, when bound to proteins, fluoresces in the infrared spectrum. Viral samples are separated by SDS–PAGE followed by Coomassie Brilliant Blue staining and gel analysis with an infrared laser-scanning device. In combination with a protein standard, our method allows the rapid and accurate determination of viral particle titers simultaneously with the assessment of vector purity. PMID:22816378

Kohlbrenner, Erik; Henckaerts, Els; Rapti, Kleopatra; Gordon, Ronald E.; Linden, R. Michael; Hajjar, Roger J.

2012-01-01

168

Apparatus for eliminating background interference in fluorescence measurements  

DOEpatents

The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser which excites a different stained component of the same biological particle.

Martin, J.C.; Jett, J.H.

1984-01-06

169

Apparatus for eliminating background interference in fluorescence measurements  

DOEpatents

The disclosure is directed to an apparatus for eliminating background interference during fluorescence measurements in a multiple laser flow cytometer. A biological particle stained with fluorescent dyes is excited by a laser. A fluorescence detector detects the fluorescence. The particle scatters light and a gate signal is generated and delayed until the biological particle reaches the next laser. The delayed signal turns on this next laser, which excites a different stained component of the same biological particle.

Martin, John C. (Los Alamos, NM); Jett, James H. (Los Alamos, NM)

1986-01-01

170

In vivo effects of focused shock waves on tumor tissue visualized by fluorescence staining techniques.  

PubMed

Shock waves can cause significant cytotoxic effects in tumor cells and tissues both in vitro and in vivo. However, understanding the mechanisms of shock wave interaction with tissues is limited. We have studied in vivo effects of focused shock waves induced in the syngeneic sarcoma tumor model using the TUNEL assay, immunohistochemical detection of caspase-3 and hematoxylin-eosin staining. Shock waves were produced by a multichannel pulsed-electrohydraulic discharge generator with a cylindrical ceramic-coated electrode. In tumors treated with shock waves, a large area of damaged tissue was detected which was clearly differentiated from intact tissue. Localization and a cone-shaped region of tissue damage visualized by TUNEL reaction apparently correlated with the conical shape and direction of shock wave propagation determined by high-speed shadowgraphy. A strong TUNEL reaction of nuclei and nucleus fragments in tissue exposed to shock waves suggested apoptosis in this destroyed tumor area. However, specificity of the TUNEL technique to apoptotic cells is ambiguous and other apoptotic markers (caspase-3) that we used in our study did not confirmed this observation. Thus, the generated fragments of nuclei gave rise to a false TUNEL reaction not associated with apoptosis. Mechanical stress from high overpressure shock wave was likely the dominant pathway of tumor damage. PMID:25200989

Lukes, Petr; Zeman, Jan; Horak, Vratislav; Hoffer, Petr; Pouckova, Pavla; Holubova, Monika; Hosseini, S Hamid R; Akiyama, Hidenori; Sunka, Pavel; Benes, Jiri

2014-08-29

171

Study of platelet behavior in vivo after endothelial stimulation with laser irradiation using fluorescence intravital videomicroscopy and PEGylated liposome staining.  

PubMed

Platelets contain an array of potent proinflammatory mediators, and therefore they are regarded as mediator and effector cells in inflammation. Knowing the role of platelets during these processes is crucial and the analysis of their behavior in situ and the associated mechanisms is consequently particularly important. However, conventional in vitro staining techniques induce modification of the characteristics of platelets. This study aimed to evaluate platelet behavior in vivo after endothelial stimulation (without endothelial denudation or exposure of basal lamina and/or collagen) with an argon laser, using video intravital microscopy in combination with a new an innovative platelet staining technique based on polyethyleneglycol (PEG) liposomes. The study was performed on skin by using a dorsal skin-fold chamber implanted in golden hamsters. Platelets were stained by 5,6-CF-encapsulated PEGylated liposomes injected intravenously. The skin microcirculation was observed with an intravital microscope (using x25, x40, and x80 magnifications) fitted with a xenon light source, an epifluorescence assembly, and an ultra-high sensitivity video camera for fluorescence imaging. Platelet activation without endothelial denudation or exposure of basal lamina and/or collagen was obtained with an argon laser emitting at 514.5 nm with the following parameters: 20 mW, 300 ms, 120 J/cm(2). The 80-microm laser beam was focused on a vessel and its position was controlled with the microscope. Thanks to the spatial resolution of the intravital microscopic imaging system, the platelets were seen rolling individually on the endothelium. After laser stimulation, platelets were activated and three phases were observed: recruitment, adhesion and detachment. The observation of these three phases was time dependent and the kinetics of the process were quantified. The recruitment reached a maximum after 90 +/- 20 s. The adhesion phase lasted for 110 +/- 25 s. At last, detachment of all platelets was observed. This detachment started 200 +/- 20 s after irradiation and was completed in less than 2 min. This study confirms that laser irradiation used with optimal parameters can induce platelet activation without thrombus formation. Platelets can adhere only transiently on stimulated endothelium. This phenomenon may therefore represent a defense mechanism, by which platelets would accumulate in the vicinity of an injury, making them available for immediate response. At last, this study has clearly demonstrated the advantages of our new and innovative platelet staining method using PEGylated liposomes, which are (i) in situ labeling, (ii) use of a hydrophilic marker located in an aqueous compartment within the platelet, and (iii) labeling of platelets allowing observation during the whole experiment. PMID:12204655

Mordon, Serge; Begu, Sylvie; Buys, Bruno; Tourne-Peteilh, Corinne; Devoisselle, J M

2002-09-01

172

Silica cross-linked nanoparticles encapsulating fluorescent conjugated dyes for energy transfer-based white light emission and porphyrin sensing.  

PubMed

This work demonstrated that water-soluble fluorescent hybrid materials can be successfully synthesized by use of silica cross-linked micellar nanoparticles (SCMNPs) as scaffolds to encapsulate fluorescent conjugated dyes for pH sensing, porphyrin sensing and tunable colour emission. Three dyes were separately encapsulated inside SCMNPs (short to dye-SCMNPs). Each of the dye-SCMNPs indicated longer lifetime in water than that of free dye dissolved in organic solvent. The 7-(hexadecyloxy) coumarin-3-ethylformate (HCE) encapsulated inside SCMNPs (HCE-SCMNPs) exhibited fluorescence quenching by pH change in aqueous media. Furthermore, it was confirmed that the radiative and nonradiative energy transfer processes both occurred between HCE-SCMNPs and tetraphenyl-porphyrin (TPP), which were used to synthesize the water-soluble TPP sensor. Significantly, HCE-SCMNPs doped with 5,12-dicotyl-quinacridone (8CQA) and TPP showed water-soluble white light emission (CIE (0.29, 0.34)) upon singlet excitation of 376 nm due to colour adjustment of 8CQA and energy transfer from HCE (donor) to TPP (acceptor). PMID:22930394

Gai, Fangyuan; Zhou, Tianlei; Zhang, Ligong; Li, Xiang; Hou, Weijia; Yang, Xinchun; Li, Yantao; Zhao, Xiaogang; Xu, Da; Liu, Yunling; Huo, Qisheng

2012-09-28

173

Characterization of the vitreous body of the human eye using a cyanine dye as a spectral and fluorescent probe  

NASA Astrophysics Data System (ADS)

We used one of cyanine dyes as a spectral and fluorescent probe in the study of the composition of the extracellular matrix of the human eye (its vitreous body). Owing to the unique ability of the dye to bind to collagens and human serum albumin, we revealed the simultaneous presence of both types of biomacromolecules in the vitreous body. The formation of the dye complex with human serum albumin leads to appearance of a long-wavelength absorption band (~612 nm) and a steep rise of fluorescence, whereas in the presence of collagens the dye forms J-aggregates with a longer-wavelength absorption band (640-660 nm) and moderate fluorescence. In this work we studied the composition of the human fetus vitreous body and its dynamics from 9 to 31 gestation weeks. On the basis of the data obtained by this method, we may assume that albumin, being a carrier protein, probably provides the vitreous body and surrounding tissues with necessary growth factors, hormones, lipids, vitamins, and some other biomolecules. The data show that the dye is promising not only for study of albumin functions in eye development, but also for characterization of some eye diseases and for analysis of other extracellular media.

Panova, Ina G.; Tatikolov, Alexander S.

2009-02-01

174

Synthesis and characterization of monodisperse, mesoporous, and magnetic sub-micron particles doped with a near-infrared fluorescent dye  

SciTech Connect

Recently, multifunctional silica nanoparticles have been investigated extensively for their potential use in biomedical applications. We have prepared sub-micron monodisperse and stable multifunctional mesoporous silica particles with a high level of magnetization and fluorescence in the near infrared region using an one-pot synthesis technique. Commercial magnetite nanocrystals and a conjugated-NIR-dye were incorporated inside the particles during the silica condensation reaction. The particles were then coated with polyethyleneglycol to stop aggregation. X-ray diffraction, N{sub 2} adsorption analysis, TEM, fluorescence and absorbance measurements were used to structurally characterize the particles. These mesoporous silica spheres have a large surface area (1978 m{sup 2}/g) with 3.40 nm pore diameter and a high fluorescence in the near infrared region at {lambda}=700 nm. To explore the potential of these particles for drug delivery applications, the pore accessibility to hydrophobic drugs was simulated by successfully trapping a hydrophobic ruthenium dye complex inside the particle with an estimated concentration of 3 wt%. Fluorescence imaging confirmed the presence of both NIR dye and the post-grafted ruthenium dye complex inside the particles. These particles moved at approximately 150 {mu}m/s under the influence of a magnetic field, hence demonstrating the multifunctionality and potential for biomedical applications in targeting and imaging. - Graphical Abstract: Hydrophobic fluorescent Ruthenium complex has been loaded into the mesopores as a surrogate drug to simulate drug delivery and to enhance the multifunctionality of the magnetic NIR emitting particles. Highlights: > Monodisperse magnetic mesoporous silica particles emitting in the near infrared region are obtained in one-pot synthesis. > We prove the capacity of such particles to uptake hydrophobic dye to mimic drug loading. > Loaded fluorescent particles can be moved under a magnetic field in a microfluidic device.

Le Guevel, Xavier, E-mail: xavier.leguevel@dcu.ie [Biomedical Diagnostics Institute, School of Physical Sciences, Dublin City University, Glasnevin, Dublin 9 (Ireland); Nooney, Robert; McDonagh, Colette; MacCraith, Brian D. [Biomedical Diagnostics Institute, School of Physical Sciences, Dublin City University, Glasnevin, Dublin 9 (Ireland)

2011-06-15

175

Fluorescent nanomicelles for selective detection of Sudan dye in Pluronic F127 aqueous media.  

PubMed

Novel self-assembled water-soluble nanomicelles that contain fluorescent conjugated polymers (poly(9,9-dioctylfluorene) (PFO) or poly[2,7-(9,9-dihexylfluorene)-alt-4,4'-phenylether] (PF-PE)) have been obtained and used as the highly sensitive/selective platform for Sudan dye detection. The Fluorescent nanomicelles exhibited a highly selective fluorescence quenching by the prohibited food additive Sudan I, while not for the natural pigments: Capsanthin and Beta-carotene, due to the more suitable matching of the LUMOs (lowest unoccupied molecular orbital) of the conjugated polymers with that of Sudan I molecules. The Stern-Volmer constants (K(SV)) of PF-PE/F127 and PFO/F127 for Sudan I were 1,040,480 and 665,000 M(-1), respectively, which were more than 100 times higher than those of the same conjugated polymers in the orgainc solvents. The significantly enhanced sensitivity was due to the collective effect of the F127 micelles to both chromophore and analyte, through which the fluorophone-analyte binding interaction is significantly strengthened and efficient photoinduced charge transfer occurs. The as-proposed materials and approach may be potentially applied in the real-time food safety screening. PMID:24625370

Ye, Xinliang; Zhang, Jie; Chen, Hui; Wang, Xiaohui; Huang, Fei

2014-04-01

176

Signal Decomposition of Transmembrane Voltage-Sensitive Dye Fluorescence Using a Multiresolution Wavelet Analysis  

PubMed Central

Fluorescence imaging of transmembrane voltage-sensitive dyes is used to study electrical activation in cardiac tissue. However, the fluorescence signals, typically, have low SNRs and may be contaminated with motion artifact. In this report, we introduce a new processing approach for fluoresced transmembrane potentials (fTmps) that is based upon a discrete wavelet transform. We show how fTmp signals can be decomposed and reconstructed to form three subsignals that contain signal noise (noise signal), the early depolarization phase of the action potential (rTmp signal), and motion artifact (rMA signal). A coiflet4 wavelet is used for fTmp decomposition and reconstruction of these subsignals. Results using fTmp signals that are contaminated with motion artifact indicate that the approach is a useful processing step to remove baseline drift, reduce noise, and reveal wavefronts. It streamlines the preprocessing of fTmps for the subsequent measurement of activation times and conduction velocities. It is a promising approach for studying wavefronts without aggressive mechanical tissue constraint or electromechanical uncoupling agents and is, useful for single-camera systems that do not provide for ratiometric imaging. PMID:21511560

Asfour, Huda; Swift, Luther M.; Sarvazyan, Narine; Doroslova?ki, Miloš; Kay, Matthew W.

2013-01-01

177

Monitoring FET flow control and wall adsorption of charged fluorescent dye molecules in nanochannels integrated into a multiple internal reflection  

E-print Network

Monitoring FET flow control and wall adsorption of charged fluorescent dye molecules in nanochannels integrated into a multiple internal reflection infrared waveguide Youn-Jin Oh,a Thomas C. Gamble fabricated multiple internal reflection infrared waveguides embedded with a parallel array of nanofluidic

New Mexico, University of

178

Single-lane single-fluor sequencing using dideoxy-labeled, heavy-atom-modified near-IR fluorescent dyes  

NASA Astrophysics Data System (ADS)

Using a near-IR (NIR) fluorescence detection system and labels synthesized in our laboratories, electropherograms of oligonucleotides separated by capillary gel electrophoresis and detected using NIR fluorescence will be presented. The sequence of nucleotide bases was determined using a single-lane, single-dye technique. The molar concentrations of the ddNTP's used during extension reactions were varied in order to achieve a ratio of 4:2:1:0 (A:C:G:T) which allowed the identification of each terminal base via fluorescence intensity measurements. Sequencing ladders were prepared from the template, M13mp18, using standard Sanger dideoxy chain termination techniques, the modified T7 DNA polymerase, and a NIR-labeled M13 primer. The data indicated reliable sequence determination up to 300 bases with a base-calling accuracy of 90%. In order to eliminate the need for dye-labeled primers and the T7 DNA polymerase enzyme, we have developed a sequencing strategy which utilizes dye-labeled dideoxy nucleotides in a single-lane, single-fluor approach. Base-calling is accomplished by measuring the fluorescence lifetime of intramolecular heavy-atom modified dyes.

Williams, Daryl C.; Flanagan, James H., Jr.; Legendre, Benjamin L., Jr.; Hammer, Robert P.; Soper, Steven A.

1995-04-01

179

Fluorescence enhancement through modified dye molecule absorption associated with the localized surface plasmon resonances of metallic dimers  

NASA Astrophysics Data System (ADS)

Nano-antennae consisting of gold particle dimers were fabricated by electron-beam lithography. Dark-field scattering spectroscopy was used to probe the plasmonic response of individual nano-antennae and to characterize the localized surface plasmon resonances they support. Fluorescence from dye molecules dispersed in a thin polymer film that covered the dimers was used to probe the interaction between fluorophores and the nano-antennae. Through a suitable choice of dye emission spectrum and spectral position of the dimer resonance, we were able to focus on the way the plasmon resonances may mediate absorption of incident light by the dye. We separated out the role of plasmon resonances on absorption from emission. This was done using energy transfer in a donor acceptor pair of dyes.

Zoriniants, George; Barnes, William L.

2008-10-01

180

Monitoring changes in membrane polarity, membrane integrity, and intracellular ion concentrations in Streptococcus pneumoniae using fluorescent dyes.  

PubMed

Membrane depolarization and ion fluxes are events that have been studied extensively in biological systems due to their ability to profoundly impact cellular functions, including energetics and signal transductions. While both fluorescent and electrophysiological methods, including electrode usage and patch-clamping, have been well developed for measuring these events in eukaryotic cells, methodology for measuring similar events in microorganisms have proven more challenging to develop given their small size in combination with the more complex outer surface of bacteria shielding the membrane. During our studies of death-initiation in Streptococcus pneumoniae (pneumococcus), we wanted to elucidate the role of membrane events, including changes in polarity, integrity, and intracellular ion concentrations. Searching the literature, we found that very few studies exist. Other investigators had monitored radioisotope uptake or equilibrium to measure ion fluxes and membrane potential and a limited number of studies, mostly in Gram-negative organisms, had seen some success using carbocyanine or oxonol fluorescent dyes to measure membrane potential, or loading bacteria with cell-permeant acetoxymethyl (AM) ester versions of ion-sensitive fluorescent indicator dyes. We therefore established and optimized protocols for measuring membrane potential, rupture, and ion-transport in the Gram-positive organism S. pneumoniae. We developed protocols using the bis-oxonol dye DiBAC4(3) and the cell-impermeant dye propidium iodide to measure membrane depolarization and rupture, respectively, as well as methods to optimally load the pneumococci with the AM esters of the ratiometric dyes Fura-2, PBFI, and BCECF to detect changes in intracellular concentrations of Ca(2+), K(+), and H(+), respectively, using a fluorescence-detection plate reader. These protocols are the first of their kind for the pneumococcus and the majority of these dyes have not been used in any other bacterial species. Though our protocols have been optimized for S. pneumoniae, we believe these approaches should form an excellent starting-point for similar studies in other bacterial species. PMID:24637356

Clementi, Emily A; Marks, Laura R; Roche-Håkansson, Hazeline; Håkansson, Anders P

2014-01-01

181

Fluorescence spectra of organic dyes in solution: a time dependent multilevel approach.  

PubMed

Classical all-atom molecular dynamics (MD) simulations and quantum mechanical (QM) time-dependent density functional theory (TD-DFT) calculations are employed to study the conformational and photophysical properties of the first emitter excited state of tetramethyl-rhodamine iso-thiocyanate fluorophore in aqueous solution. For this purpose, a specific and accurate force field has been parameterised from QM data to model the fluorophore's first bright excited state. During the MD simulations, the consequences of the ???* electronic transition on the structure and microsolvation sphere of the dye has been analysed in some detail and compared to the ground state behaviour. Thereafter, fluorescence has been calculated at the TD-DFT level on configurations sampled from the simulated MD trajectories, allowing us to include time dependent solvent effects in the computed emission spectrum. The latter, when compared with the absorption spectrum, reproduces well the experimental Stokes shift, further validating the proposed multilevel computational procedure. PMID:21127788

Barone, Vincenzo; Bloino, Julien; Monti, Susanna; Pedone, Alfonso; Prampolini, Giacomo

2011-02-14

182

Carboxylate-modified squaraine dye doped silica fluorescent pH nanosensors.  

PubMed

Novel carboxylate-modified fluorescent silica pH nanosensors were synthesized using a reverse microemulsion method with a pH sensitive squaraine dye used as pH indicator. This pH sensitive squaraine dye was simply doped inside SiNPs without any complicated procedures. To avoid aggregation among the particles and to increase the water solubility of the pH nanosensors, the SiNPs were surface modified with a carboxyl group. This pH probe exhibits a good linear dynamic response between pH 3.01 and 5.72. Many alkali, alkaline earth, and transitional metal ions including Li( + ), Na( + ), K( + ), Rb( + ), Cs( + ), Mg(2 + ), Ca(2 + ), Sr(2 + ), Al(3 + ), V(5 + ), Cr(3 + ), Cr(6 + ), Mn(2 + ), Fe(2 + ), Fe(3 + ), Co(2 + ), Ni(3 + ), Cu(2 + ), Zn(2 + ), As(3 + ), Se(4 + ), Mo(6 + ), Ag( + ), Cd(2 + ), La(3 + ), Er(3 + ), Ir(3 + ), Hg( + ), Hg(2 + ), and Pb(2 + ) had no significant interference on pH value determination. Artificial sample determination showed that the pH nanosensors developed in this work possess a very promising applicability in biological and biomedical fields. PMID:20431191

Xue, Lina; Li, Baiyan; Fei, Qiang; Feng, Guodong; Huan, Yanfu; Shi, Zhan

2010-05-28

183

Total chemical synthesis of biologically active fluorescent dye-labeled Ts1 toxin.  

PubMed

Ts1 toxin is a protein found in the venom of the Brazilian scorpion Tityus serrulatus. Ts1 binds to the domain II voltage sensor in the voltage-gated sodium channel Nav and modifies its voltage dependence. In the work reported here, we established an efficient total chemical synthesis of the Ts1 protein using modern chemical ligation methods and demonstrated that it was fully active in modifying the voltage dependence of the rat skeletal muscle voltage-gated sodium channel rNav1.4 expressed in oocytes. Total synthesis combined with click chemistry was used to label the Ts1 protein molecule with the fluorescent dyes Alexa-Fluor 488 and Bodipy. Dye-labeled Ts1 proteins retained their optical properties and bound to and modified the voltage dependence of the sodium channel Nav. Because of the highly specific binding of Ts1 toxin to Nav, successful chemical synthesis and labeling of Ts1 toxin provides an important tool for biophysical studies, histochemical studies, and opto-pharmacological studies of the Nav protein. PMID:24989851

Dang, Bobo; Kubota, Tomoya; Correa, Ana M; Bezanilla, Francisco; Kent, Stephen B H

2014-08-18

184

Spectroscopic characterization of diverse amyloid fibrils in vitro by the fluorescent dye Nile red.  

PubMed

The fluorescence of Nile red (9-diethylamino-5H-benzophenoxazine-5-one) is quenched in aqueous solutions but shows augmented fluorescence in hydrophobic environments. Nile red fluorescence was blue shifted and strongly augmented in the presence of various amyloid fibrils assayed under acidic as well as neutral pH conditions. Fibrils grown from lysozyme and insulin (at pH 1.6 and 65 °C), transthyretin (TTR) fibrils grown from the acid unfolded monomer (pH 2.0, 21 °C) or from the dissociated tetramer starting from native protein under less acidic conditions (pH 4.4, 37 °C) were detected. Nile red was also successfully employed in detecting A?1-42 and human prion protein (PrP90-231) amyloid fibrils grown at neutral pH. Nile red was amyloid fibril specific and did not fluoresce appreciably in the presence of the monomeric precursor proteins. Stoke's shifts of the wavelength maximum of Nile red bound to various fibrils were different (ranging from 615 nm to 638 nm) indicating sensitivity to the tertiary structure in its respective binding sites of different amyloid proteins. A polarity assay using ethanol-water mixtures and pure octanol ranging from dielectric constants between 10 and 70 showed a linear correlation of Nile red Stoke's shift and allowed assignment of amyloid fibril binding site polarity. Fluorescence resonance energy transfer between Thioflavin T (ThT) and Nile red was proven to be efficient and co-staining was employed to discriminate between conformational isoforms of A?1-42 amyloid fibrils grown under agitated and quiescent conditions. This paper demonstrates the complementary use of this fluorometric method for conformational typing of amyloid structures. PMID:21279219

Mishra, Rajesh; Sjölander, Daniel; Hammarström, Per

2011-04-01

185

Fluorescence enhancement of dyes embedded in nanoparticles of Lu, Eu, Al, and Sc diketonates of different composition and concentration  

NASA Astrophysics Data System (ADS)

We have studied the effect of central ions (Lu(III), Eu(III), Sc(III), and Al(III)), organic ligands (2-naphthoyltrifluoroacetone (NTA) and p-phenylbenzoyltrifluoroacetone (PhBTA)), and their concentration in a water-alcohol solution on the fluorescence of ?-diketonate complexes formed and nanoparticles (NPs) generated by the self-assembly of these complexes. The fluorescence quenching of ligands of the complexes of nanoparticles because of the introduction of molecules of dyes, such as Nile Blue (NB), Lissamine Rhodamine RB-200 (RB), and Crystal Violet (CV), in these nanoparticles is investigated, and the NP-sensitization of the fluorescence of these dyes is explored. The dependence of the intensity of the NP-sensitized fluorescence of NB on its concentration in nanoparticles consisting of complexes that differ in composition and concentration is studied. By analyzing this dependence for the nanoparticles consisting of Sc(NTA)3, the size of the studied nanoparticles is evaluated. It is shown that the nature of this dependence is determined by a competition of two processes: the migration of the excitation energy over complexes to dyes and the migration of the excitation energy of dyes to impurities or dimer of dyes. The size of nanoparticles is compared to the estimated values of the exciton diffusion length and the critical radius of energy transfer from complexes to NB. An energy transfer of close to 100% from the nanoparticles formed of 10 ?M of Sc(NTA)3 to 50 nM of NB molecules embedded therein is observed. The introduction of NB molecules into nanoparticles leads to a 200-fold increase in fluorescence intensity compared to their direct excitation in solution.

Mironov, L. Yu.; Sveshnikova, E. B.; Ermolaev, V. L.

2014-12-01

186

A universal fluorescent aptasensor based on AccuBlue dye for the detection of pathogenic bacteria.  

PubMed

We report a universal fluorescent aptasensor based on the AccuBlue dye, which is impermeant to cell membranes, for the detection of pathogenic bacteria. The sensor consists of AccuBlue, an aptamer strand, and its complementary strand (cDNA) that partially hybridizes to the aptamer strand. We have fabricated two models by changing the sequence of the reaction between the elements in the system. One is the "signal on" model in which the aptamer is first bound to the target, followed by the addition of cDNA and AccuBlue, at which time the cDNA hybridizes with the free unreacted aptamer and forms a double-stranded DNA (dsDNA) duplex. Such hybridization causes AccuBlue to insert into the dsDNA and exhibit significantly increased fluorescence intensity because of the specific intercalation of the AccuBlue into dsDNA rather than single-stranded DNA (ssDNA). The other model, "signal off," involves hybridization of the aptamer with cDNA first, resulting in high fluorescence intensity on the addition of AccuBlue. When the target is added, the aptamer binds the target, causing the cDNA to detach from the dsDNA duplex and resulting in low fluorescence as a result of the liberation of AccuBlue. Because this design is based purely on DNA hybridization, and AccuBlue is impermeant to cell membranes, it could potentially be adapted to a wide variety of analytes. PMID:24650583

Duan, Nuo; Wu, Shijia; Ma, Xiaoyuan; Xia, Yu; Wang, Zhouping

2014-06-01

187

High-resolution liquid chromatography of fluorescent dye-labeled nucleic acids.  

PubMed

Using 100 mM of triethylammonium acetate as ion-pairing reagent, phosphodiester oligonucleotides labeled fluorescently at their 5' terminus could be separated successfully on alkylated nonporous 2.3-microns poly(styrene-divinylbenzene) particles by means of high-resolution liquid chromatography. Applying excitation wavelengths of 490, 520, 550, and 575 nm, respectively, optimum sensitivity was achieved for the fluorophores 5-carboxyfluorescein, 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein, N,N,N',N'-tetramethyl-6-carboxyrhodamine, and 6-carboxy-X-rhodamine (FAM, JOE, TAMRA, and ROX, respectively) at emission wavelengths of 520, 550, 580, and 605 nm, respectively. With calibration curves being linear over at least three orders of magnitude, the lower detection limits were 0.5, 2, 2, and 3 fmol, respectively. Depending on the type of fluorescent dye attached, retention times increased in the order JOE < FAM < TAMRA < ROX. Subsequently, fluorescent oligonucleotides were employed to prime polymerase chain reactions (PCR). Again the fluorophores were found to increase the retention times of double-stranded nucleic acids, but to a lesser degree than those of single-stranded oligonucleotides. Using a single FAM label attached to one of the two PCR primers, the sensitivity of fluorescence detection was found to be approximately 1 fmol or 30-70 times higher than that of uv absorbance detection depending on the length of the PCR product. Since the technique allows the separation of PCR products differing only 4 to 8 base pairs in length within a size range of 50 to 200 base pairs, it may be employed for the quantitative assessment of competitive PCR. PMID:7695100

Oefner, P J; Huber, C G; Umlauft, F; Berti, G N; Stimpfl, E; Bonn, G K

1994-11-15

188

Substituent and Solvent Effects on Excited State Charge Transfer Behavior of Highly Fluorescent Dyes Containing Thiophenylimidazole-Based Aldehydes  

NASA Technical Reports Server (NTRS)

The excited state charge transfer for a series of highly fluorescent dyes containing thiophenylimidazole moiety was investigated. These systems follow the Twisted Intramolecular Charge Transfer (TICT) model. Dual fluorescence was observed for each substituted dye. X-ray structures analysis reveals a twisted ground state geometry for the donor substituted aryl on the 4 and 5 position at the imidazole ring. The excited state charge transfer was modeled by a linear solvation energy relationship using Taft's pi and Dimroth's E(sub T)(30) as solvent parameters. There is linear relation between the energy of the fluorescence transition and solvent polarity. The degree of stabilization of the excited state charge transfer was found to be consistent with the intramolecular molecular charge transfer. Excited dipole moment was studied by utilizing the solvatochromic shift method.

Santos, Javier; Bu, Xiu R.; Mintz, Eric A.

2001-01-01

189

Fluorescence microscopy is superior to polarized microscopy for detecting amyloid deposits in Congo red-stained trephine bone marrow biopsy specimens.  

PubMed

The classic gold standard for detecting amyloid deposits is Congo red-stained bright field and polarized microscopy (CRPM). A prior study showed that Congo red fluorescence (CRF) microscopy had increased sensitivity compared with traditional CRPM when analyzing fat pad specimens. The purpose of the current study was to determine the sensitivity of CRF for evaluating Congo red-stained bone marrow biopsy specimens, and to compare these results with those of CRPM. We compared the CRPM and the CRF analyses of 33 trephine bone marrow biopsy specimens with clinical or morphologic suspicion of amyloid deposits. These results were verified against immunohistochemical staining with anti-amyloid P antibody. CRF achieved 100% sensitivity, and CRPM achieved 75% sensitivity. Both groups showed 100% specificity compared with amyloid P immunohistochemical staining. The results show that CRF is a sensitive method to analyze trephine bone marrow biopsy specimens for amyloid deposits. PMID:23010714

Marcus, Alan; Sadimin, Evita; Richardson, Maurice; Goodell, Lauri; Fyfe, Billie

2012-10-01

190

Amyloid Histology Stain for Rapid Bacterial Endospore Imaging ? †  

PubMed Central

Bacterial endospores are some of the most resilient forms of life known to us, with their persistent survival capability resulting from a complex and effective structural organization. The outer membrane of endospores is surrounded by the densely packed endospore coat and exosporium, containing amyloid or amyloid-like proteins. In fact, it is the impenetrable composition of the endospore coat and the exosporium that makes staining methodologies for endospore detection complex and challenging. Therefore, a plausible strategy for facile and expedient staining would be to target components of the protective surface layers of the endospores. Instead of targeting endogenous markers encapsulated in the spores, here we demonstrated staining of these dormant life entities that targets the amyloid domains, i.e., the very surface components that make the coats of these species impenetrable. Using an amyloid staining dye, thioflavin T (ThT), we examined this strategy. A short incubation of bacillus endospore suspensions with ThT, under ambient conditions, resulted in (i) an enhancement of the fluorescence of ThT and (ii) the accumulation of ThT in the endospores, affording fluorescence images with excellent contrast ratios. Fluorescence images revealed that ThT tends to accumulate in the surface regions of the endospores. The observed fluorescence enhancement and dye accumulation, coupled with the sensitivity of emission techniques, provide an effective and rapid means of staining endospores without the inconvenience of pre- or posttreatment of samples. PMID:21653779

Xia, Bing; Upadhyayula, Srigokul; Nuñez, Vicente; Landsman, Pavel; Lam, Samuel; Malik, Harbani; Gupta, Sharad; Sarshar, Mohammad; Hu, Jingqiu; Anvari, Bahman; Jones, Guilford; Vullev, Valentine I.

2011-01-01

191

Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry  

PubMed Central

Summary We developed a rapid detection method for Legionella pneumophila (Lp) by filtration, immunomagnetic separation, double fluorescent staining, and flow cytometry (IMS?FCM method). The method requires 120?min and can discriminate ‘viable’ and ‘membrane?damaged’ cells. The recovery is over 85% of spiked Lp SG 1 cells in 1?l of tap water and detection limits are around 50 and 15 cells per litre for total and viable Lp, respectively. The method was compared using water samples from house installations in a blind study with three environmental laboratories performing the ISO 11731 plating method. In 53% of the water samples from different taps and showers significantly higher concentrations of Lp were detected by flow cytometry. No correlation to the plate culture method was found. Since also ‘viable but not culturable’ (VNBC) cells are detected by our method, this result was expected. The IMS?FCM method is limited by the specificity of the used antibodies; in the presented case they target Lp serogroups 1–12. This and the fact that no Lp?containing amoebae are detected may explain why in 21% of all samples higher counts were observed using the plate culture method. Though the IMS?FCM method is not yet fit to completely displace the established plating method (ISO 11731) for routine Lp monitoring, it has major advantages to plating and can quickly provide important insights into the ecology of this pathogen in water distribution systems. PMID:23062200

Keserue, Hans?Anton; Baumgartner, Andreas; Felleisen, Richard; Egli, Thomas

2012-01-01

192

Oxidative chemistry of fluorescent dyes: implications in the detection of reactive oxygen and nitrogen species.  

PubMed

HE (hydroethidine), a widely used fluorescent dye for detecting intracellular superoxide, undergoes specific oxidation and hydroxylation reactions. The reaction between HE and O2•- (superoxide radical) yields a diagnostic marker product, 2-hydroxyethidium. This is contrary to the popular notion that O2•- oxidizes HE to form ethidium. HE, however, undergoes a non-specific oxidation to form ethidium in the presence of other oxidants (hydroxyl radical, peroxynitrite and perferryl iron) and other dimeric products. The mitochondria-targeted HE analogue Mito-SOX® undergoes the same type of oxidative chemistry to form products similar to those formed from HE. On the basis of the oxidative chemical mechanism of HE and Mito-SOX®, we conclude that flurorescence microscopy or related techniques are not sufficient to measure the superoxide-specific hydroxylated products. HPLC methodologies are required to separate and identify these products. Peroxynitrite reacts rapidly and stoichiometrically with boronates to form specific products. Assays using fluorescent-based boronate probes will be more reliable for peroxynitrite determination than those using either dichlorodihydrofluorescein or dihydrorhodamine. PMID:21936793

Kalyanaraman, Balaraman

2011-10-01

193

Fluorescent dye-labelled polymer synthesis by nitroxide mediated radical polymerization  

NASA Astrophysics Data System (ADS)

New applications of polymers at advanced technologies demand increased requirements on their properties. These properties are influenced by molecular as well as supramolecular structure. Controlled radical polymerization mediated by stable nitroxides (NMP) or substituted alkoxyamines offers simple method for preparation of polymers with programmable structure of macromolecules which possess remarkable better physical as well as chemical properties. They can be used as a macro initiators for the synthesis of block copolymers. At the present time it has been generally accepted that the extent of "livingness" is high for all conversions [1-4]. To verify this statement a series of fluorescent dye-labelled regulators has been synthesized, spectrally characterized and used as the mediators of styrene and n-butyl acrylate polymerization. Direct quantification of dormant species concentration (extent of livingness) and calculation of molar mass of marked polymers was performed by absorption and/or emission spectroscopy. Controlled radical polymerization mediated by stable nitroxides bearing fluorescence mark represents unconventional approach for monitoring and evaluation of mechanism and kinetics of polymerization process. Results indicate that the extent of livingness is strongly influenced by conversion as well as mediator concentration. There is a clear tendency toward to decreasing amount of dormant species with increasing monomer conversion. Moreover, lower mediator concentration decreases livingness of polymerization process.

Kollár, Jozef; Chmela, Štefan; Hr?ková, Ľudmila; Hrdlovi?, Pavol

2012-07-01

194

Ex vivo imaging of excised tissue using vital dyes and confocal microscopy.  

PubMed

Vital dyes routinely used for staining cultured cells can also be used to stain and image live tissue slices ex vivo. Staining tissue with vital dyes allows researchers to collect structural and functional data simultaneously and can be used for qualitative or quantitative fluorescent image collection. The protocols presented here are useful for structural and functional analysis of viable properties of cells in intact tissue slices, allowing for the collection of data in a structurally relevant environment. With these protocols, vital dyes can be applied as a research tool to disease processes and properties of tissue not amenable to cell culture-based studies. PMID:22752953

Johnson, Simon; Rabinovitch, Peter

2012-07-01

195

Use of Fluorescence Imaging in Combination with Patent Blue Dye versus Patent Blue Dye Alone in Sentinel Lymph Node Biopsy in Breast Cancer  

PubMed Central

Purpose Near-infrared fluorescence imaging with indocyanine green (ICG) has the potential to improve sentinel lymph node (SLN) mapping in breast cancer. In this clinical trial, we compared the potential value of ICG combined with blue dye with that of blue dye alone for detecting SLNs. Methods Patients undergoing SLN biopsy (SLNB) between November 2010 and November 2013 were included. Up to December 2011, SLNs were detected by using patent blue (PB) alone, and since January 2012, by using PB in combination with ICG. The patients were divided into the following two groups: group A (ICG-PB; n=96) and group B (PB; n=73), and SLN detection parameters were compared between the groups. All patients underwent level I and II axillary dissections after SLNB. Results In group A, the SLN detection rate was 96.9% (93/96), the accuracy of detection was 98.9% (92/93), and the false-negative rate (FNR) was 3.4% (1/29). In group B, the SLN detection rate was 84.9% (62/73), the accuracy of detection was 96.8% (60/62), and the FNR was 11.1% (2/18). The ICG-PB group showed significantly superior results compared to the PB group for SLN detection (p=0.005) and a greatly improved FNR. Conclusion The combined fluorescence and blue dye-based tracer technique was superior to the use of blue dye alone for identifying SLNs, and for predicting axillary lymph node status in patients with breast cancer; in addition, the combined technique had reduced false-negative results. PMID:25320623

Tong, Meng; Gao, Wei

2014-01-01

196

SELECTIVITY AND SPECIFICITY OF SMALL MOLECULE FLUORESCENT DYES/PROBES USED FOR THE DETECTION OF Zn2+ AND Ca2+ IN CELLS  

PubMed Central

Fluorescent dyes are widely used in the detection of labile (free or exchangeable) Zn2+ and Ca2+ in living cells. However, their specificity over other cations and selectivity for detection of labile vs. protein-bound metal in cells remains unclear. We characterized these important properties for commonly used Zn2+ and Ca2+ dyes in a cellular environment. By tracing the fluorescence emission signal along with UV-Vis and size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS) in tandem, we demonstrated that among the dyes used for Zn2+, Zinpyr-1 fluoresces in the low molecular mass (LMM) region containing labile Zn2+, but also fluoresces in different molecular mass regions where zinc ion is detected. However, FluoZin™-3 AM, Newport Green™ DCF and Zinquin ethyl ester display weak fluorescence, lack of metal specificity and respond strongly in the high molecular mass (HMM) region. Four Ca2+ dyes were studied in an unperturbed cellular environment, and two of these were tested for binding behavior under an intracellular Ca2+ release stimulus. A majority of Ca2+ was in the labile form as tested by SEC-ICP-MS, but the fluorescence from Calcium Green-1™ AM, Oregon Green® 488 BAPTA-1, Fura red™ AM and Fluo-4 NW dyes in cells did not correspond to free Ca2+ detection. Instead, the dyes showed non-specific fluorescence in the mid- and high-molecular mass regions containing Zn, Fe and Cu. Proteomic analysis of one of the commonly seen fluorescing regions showed the possibility for some dyes to recognize Zn and Cu bound to metallothionein-2. These studies indicate that Zn2+ and Ca2+ binding dyes manifest fluorescence responses that are not unique to recognition of labile metals and bind other metals, leading to suboptimal specificity and selectivity. PMID:24356796

Landero-Figueroa, Julio A.; Vignesh, Kavitha Subramanian; Deepe, George; Caruso, Joseph

2014-01-01

197

Fluorescent stain method for the simultaneous determination of mitochondrial potential and integrity of plasma and acrosomal membranes in boar sperm.  

PubMed

The purpose of this study was to validate a technique for simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in boar spermatozoa, using an association of fluorescent probes: Propidium iodide (PI), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and JC-1. Three ejaculates from each of four different boars, all showing motility >or=80% and abnormal morphology stain technique. To a 150-microl aliquot of diluted semen it was added 3 microl of PI (0.5 mg/ml), 2 microl of JC-1 (153 microm) and 50 microl of FITC-PSA (100 microg/ml). Samples were incubated at 38.5 degrees C for 8 min, in the dark. An 8-microl sample was put on a slide, coverslipped and immediately evaluated by epifluorescent microscopy. The association of fluorescent probes was divided into eight cell classes, according to plasma membrane integrity, intact acrosome and mitochondrial function. For plasma membrane integrity, detected by PI probe, the equation: (p < 0.0001) and R(2) = 0.97 was obtained. The intact acrosome, verified by the FITC-PSA probe, produced the equation: (p < 0.0001) and R(2) = 0.98. The mitochondrial potential, marked by JC-1, was estimated by the equation: (p < 0.001) and R(2) = 0.99. The group of spermatozoa with combined intact plasma membrane, intact acrosome and high mitochondrial potential (IPIAH), was estimated by the equation: (p < 0.0001) and R(2) = 0.97. The resulting linear equations demonstrate that this technique is efficient and practical for the simultaneous evaluations of the plasma, acrosomal and mitochondrial membranes in boar spermatozoa. PMID:17348977

de Andrade, A F C; de Arruda, R P; Celeghini, E C C; Nascimento, J; Martins, S M M K; Raphael, C F; Moretti, A S

2007-04-01

198

Directional Fluorescence Spectra of Laser Dye in Opal and Inverse Opal Photonic Crystals Lydia Bechger,* Peter Lodahl, and Willem L. Vos  

E-print Network

Directional Fluorescence Spectra of Laser Dye in Opal and Inverse Opal Photonic Crystals Lydia polystyrene opals and alumina inverse opals are studied, allowing us to compare direct and inverted structures emission was first reported in refs 6 and 7: titania inverse opals doped with laser dye showed a broadband

Vos, Willem L.

199

Red/blue spectral shifts of laser-induced fluorescence emission due to different nanoparticle suspensions in various dye solutions.  

PubMed

Red/blue shifts of laser-induced fluorescence (LIF) are investigated using several guest dielectric nanoscatterers, such as TiO2, ZnO, Al2O3, and SiO2, in the host Rd6G, RdB, Coumarin 4, and Coumarin 7 ethanolic solutions. A couple of inflection points are identified varying nanoparticle (NP) density into dye solutions based on LIF spectroscopy. The inflection of the spectral shift exhibits that the suspension of NPs in dye solutions significantly involves a couple of competitive chemical and optical mechanisms during photon traveling in scattering media regarding ballistic and diffusive transport. It is shown that the low, medium, and high NP additives in fluorescent suspension induce blue, red, and blue spectral shifts, respectively. PMID:25321111

Bavali, A; Parvin, P; Mortazavi, S Z; Mohammadian, M; Mousavi Pour, M R

2014-08-20

200

Bioconjugatable azo-based dark-quencher dyes: synthesis and application to protease-activatable far-red fluorescent probes.  

PubMed

We describe the efficient synthesis and one-step derivatization of novel, nonfluorescent azo dyes based on the Black Hole Quencher-3 (BHQ-3) scaffold. These dyes were equipped with various reactive and/or bioconjugatable groups (azido, ?-iodoacetyl, ketone, terminal alkyne, vicinal diol). The azido derivative was found to be highly reactive in the context of copper-catalyzed azide-alkyne cycloaddition (CuAAC) reactions and allowed easy synthetic access to the first water-soluble (sulfonated derivative) and aldehyde-modified BHQ-3 dyes, the direct preparation of which failed by means of conventional azo-coupling reactions. The aldehyde- and ?-iodoacetyl-containing fluorescence quenchers were readily conjugated to aminooxy- and cysteine-containing peptides by the formation of a stable oxime or thioether linkage, respectively. Further fluorescent labeling of the resultant peptide conjugates with red- or far-red-emitting rhodamine or cyanine dyes through sequential and/or one-pot bioconjugations, led to novel Förster resonance energy transfer (FRET) based probes suitable for the in vivo detection and imaging of urokinase plasminogen activator, a key protease in cancer invasion and metastasis. PMID:23255474

Chevalier, Arnaud; Massif, Cédrik; Renard, Pierre-Yves; Romieu, Anthony

2013-01-28

201

Interaction of fluorescence dyes with 5-fluorouracil: A photoinduced electron transfer study in bulk and biologically relevant water  

NASA Astrophysics Data System (ADS)

The interactions of widely used chemotherapeutic drug, 5-fluorouracil (5FU) with coumarin dyes have been investigated for the first time using steady-state and time-resolved fluorescence spectroscopic measurements. The fluorescence quenching along with the decrease in lifetimes of excited state of coumarin derivatives with gradual addition of 5FU is explained by photoinduced electron transfer (PET) mechanism. Our studies were performed in bulk water and confined water of AOT (aerosol OT) reverse micelle to investigate the effect of confinement on PET dynamics. The feasibility of PET reaction for coumarin-5FU systems is investigated calculating the standard free energy changes using the Rehm-Weller equation.

Kuchlyan, Jagannath; Banik, Debasis; Kundu, Niloy; Roy, Arpita; Sarkar, Nilmoni

2014-10-01

202

A multifunctional magnetic nanocarrier bearing fluorescent dye for targeted drug delivery by enhanced two-photon triggered release  

Microsoft Academic Search

We report a novel nanoformulation for targeted drug delivery which utilizes nanophotonics through the fusion of nanotechnology with biomedical application. The approach involves an energy-transferring magnetic nanoscopic co-assembly fabricated of rhodamine B (RDB) fluorescent dye grafted gum arabic modified Fe3O4 magnetic nanoparticle and photosensitive linker by which dexamethasone drug is conjugated to the magnetic nano-assembly. The advantage offered by this

Shashwat S. Banerjee; Dong-Hwang Chen

2009-01-01

203

Dynamics of the higher lying excited states of cyanine dyes. An ultrafast fluorescence study.  

PubMed

The electronic relaxation dynamics of the second singlet excited states of several cyanine dyes was studied through the femtosecond fluorescence up-conversion technique. Our interest in these molecules comes from the potential applications of systems with upper excited singlet states with a long lifetime, which can include electron and energy transfer from the higher lying singlets after one- or two-photon absorption. We studied three series of cyanines with 4-quinolyl, 2-quinolyl, or benzothiazolyl type end groups, each with varying sp(2) carbon conjugation lengths in the methinic bridge. The dynamics after electronic excitation to singlet states above the fluorescent state vary significantly as a function of cyanine structure and conjugation length. In particular, for the 4-quinolyl series the cyanine with an intermediate conjugation length (three methinic carbons) has the slowest S2 decays with lifetimes of 5.4 ps in ethanol and 6.6 ps in ethylene glycol. On the other hand, we observed that the 2-quinolyl family has S2 decay times in the subpicosecond range independent of the conjugation length between the end groups. The slowest internal conversion was observed for the benzothiazolyl type cyanine with five methinic carbons, with an S2 lifetime of 17.3 ps in ethanol. For the planar cyanines of this study we observed for the first time a clear systematic trend in the S2 decay times which closely follow the energy gap law. It was also demonstrated that a slow S2 decay is as well observed upon excitation through degenerate two-photon absorption with near-IR pulses. The present study isolates the most important variables for the design of cyanines with long S2 lifetimes. PMID:23697505

Guarin, Cesar A; Villabona-Monsalve, Juan P; López-Arteaga, Rafael; Peon, Jorge

2013-06-20

204

Use of DNA stains in immunophenotyping by slide-based cytometry  

NASA Astrophysics Data System (ADS)

Immunophenotyping of peripheral blood leukocytes (PBL) is a very well documented application of Slide Based Cytometry (SBC). As for any other assay it is of highest importance to ensure that all cells which are relevant for an analysis are recognized. Unlike assays for cultured cells which have homogenous morphology immunophenotyping of PBLs is performed on cells with heterogeneous size and shape. Therefore, triggering on parameters related to cell morphology might lead to an incomplete analysis of just a subset of cells especially in pathological conditions. Several dyes stain DNA specifically in a wide variety of emission spectra. Many of them show some influence of the chromatin condensation and organization on the staining intensity. DNA dyes therefore can be used to differentiate between cell types having the same ploidy. This can be exploited for immunophenotyping since some dyes therefore can partially replace antibody staining. The concept of using DNA dyes in the setting of immunostaining has the following advantages: (1) nuclear staining provides a stable and easy triggering signal that guarantees both, that neither cells are excluded nor that debris or polluting particles are included into the analysis; (2) some DNA dyes differentiate between mononuclear and polymorphonuclear cells. A disadvantage of DNA dyes is that mostly cells have to be permeabilized. Because of this only one set of immunophenotypic markers can be stained, cells are fixed and permeabilized, and then nuclei are stained with the appropriate DNA dye. In the study we demonstrate the use of the most commonly available DNA dyes (7-AAD, To-Pro, To-To, PI etc.) in their applicability in immunophenotyping. An overview of spectral properties, fluorescence spill-over and optimal combinations with surface antigen staining will be shown. As in general for SBC only very small sample volumes are needed. This allows to serially analyze PBL in clinical settings that up to now could not be studied in detail such as in the critical ill patient, during major surgery, and in new-borns and infants.

Gerstner, Andreas O. H.; Laffers, Wiebke; Bootz, Friedrich; Tarnok, Attila

2003-06-01

205

Treatment of port wine stains with pulsed dye laser: a retrospective study of 848 cases in Shandong Province, People’s Republic of China  

PubMed Central

Background Currently, 595 nm pulsed dye laser (PDL) therapy is offered as one of the effective treatments of port wine stains (PWSs). However, the efficacy of PDL differs in different populations. Objective The purpose of the study was to investigate the efficacy, and related factors, of 595 nm PDL in the treatment of PWSs in Chinese patients with skin type III to IV. Methods A total of 848 cases that were treated with PDL were enrolled and analyzed in this study. An independent dermatologist evaluated these lesions according to the before and after photographs. Results The response rate (RR) of all the 848 PWS patients was 69.9%, within which the cure rate was 6.3%. The patients aged ?1 year had the highest RR (93.9%), whereas those treated after age 50 reacted the worst (RR =25%). We analyzed the anatomical distribution of the lesion and found that the temporal region had the highest lesion clearance (RR =75.3%), while the extremities had the lowest clearance (RR =44.5%). Compared with the patients whose lesion size was larger than 80 cm2, the patients with small lesion size, of 0–20 cm2, had better clinical effect (RR =73.8% vs 53.2%). The reactions of the patients with hyperplastic lesion were worse than those with red patches (RR =36.4% vs 71.7%). As well, increasing treatment numbers could achieve higher clearance rates (P=0.005). Conclusion The PDL had a relatively high RR but a low clearance rate in Chinese patients with PWS, although the earlier the intervention, the better was the efficacy. The response of PDL was, not only related to the anatomical area, but also, to the lesion size, type of lesion (ie, the presence of existing hyperplastic lesions), and the number of treatment, all of which are essential for the evaluation of therapeutic effect and acquisition of patients consent before treatment. PMID:25548515

Shi, Wenhao; Wang, Jinliang; Lin, Yan; Geng, Jianhui; Wang, Haixia; Gong, Yueqin; Liu, Huaxu; Zhang, Furen

2014-01-01

206

Fluorescent Porous Film Modified Polymer Optical Fiber via "Click" Chemistry: Stable Dye Dispersion and Trace Explosive Detection.  

PubMed

In this paper, we report a facile strategy to fabricate fluorescent porous thin film on the surface of U-bent poly(methyl methacrylate) optical fiber (U-bent POF) in situ via "click" polymerization for vapor phase sensing of explosives. Upon irradiation of evanescent UV light transmitting within the fiber under ambient condition, a porous film (POSS-thiol cross-linking film, PTCF) is synthesized on the side surface of the fiber by a thiol-ene "click" reaction of vinyl-functionalized polyhedral oligomeric silsesquioxanes (POSS-V8) and alkane dithiols. When vinyl-functionalized porphyrin, containing four allyl substituents at the periphery, is added into precursors for the polymerization, fluorescence porphyrin can be covalently bonded into the cross-linked network of PTCF. This "fastened" way reduces the aggregation-induced fluorescence self-quenching of porphyrin and enhances the physicochemical stability of the porous film on the surface of U-bent POF. Fluorescent signals of the PTCF/U-bent POF probe made by this method exhibit high fluorescence quenching toward trace TNT and DNT vapor and the highest fluorescence quenching efficiency is observed for 1, 6-hexanedimercaptan-based film. In addition, because of the presence of POSS-V8 with multi cross-linkable groups, PTCF exhibits well-organized pore network and stable dye dispersion, which not only causes fast and sensitive fluorescence quenching against vapors of nitroaromatic compounds, but also provides a repeatability of the probing performance. PMID:25487515

Ma, Jiajun; Lv, Ling; Zou, Gang; Zhang, Qijin

2015-01-14

207

Native chemical ligation combined with spirocyclization of benzopyrylium dyes for the ratiometric and selective fluorescence detection of cysteine and homocysteine.  

PubMed

Spirocyclization of xanthene dyes has become a powerful technique for developing fluorescent probes. Herein, we extend this unique fluorescence switching mechanism to a near-infrared (NIR) dye, 2-(7-diethylamino-2-oxo-2H-1-benzopyran-3-yl)-4-(2-carboxyphenyl)-7-diethylamino-1-benzopyrylium (CB), and construct a ratiometric fluorescent probe 1 for cysteine (Cys)/homocysteine (Hcy). The ratiometric sensing of probe 1 toward Cys/Hcy is realized by utilizing a tandem native chemical ligation/spirocyclization reaction to interrupt the large ?-conjugated system of CB fluorophore, thereby affording remarkable blue shifts in the spectra of sensing system (from 669 to 423 nm in absorption spectra and from 694 to 474 nm in emission spectra). Probe 1 shows a high sensitivity for Cys/Hcy, and the detection limits (3 ?) for Cys and Hcy are 1.6 × 10(-7) and 1.8 × 10(-7) M, respectively. Moreover, since both the sulfhydril and the adjacent amino groups are involved in the sensing process, probe 1 is selective toward Cys/Hcy over other thiols such as glutathione. All these unique features make it particularly favorable for ratiometric Cys/Hcy sensing and bioimaging applications. It has been preliminarily used for Cys detection in rabbit serum samples and the ratiometric fluorescent imaging of Cys in living HepG2 cells. PMID:24410246

Lv, Hongmin; Yang, Xiao-Feng; Zhong, Yaogang; Guo, Yuan; Li, Zheng; Li, Hua

2014-02-01

208

[Observation of double-stained African green monkey kidney COS-7 cells using total internal reflection double-channel fluorescence microscopy].  

PubMed

Using a Dual-View wavelength splitter, double-stained African green monkey kidney COS-7 cells, transfected with pEGFP-Myosin 15a and costained with Rhodamine-filopodia were observed based on an ICCD(intensified charge couple device) fluorescence micro-imaging systems. Total internal reflection fluorescence microscopy was used to observe the overexpression of Myosin 15a to the tips of the elongation filopodia. An approach to collecting fluorescence in two channels and avoiding spectra crosstalk was employed to observe Myosin 15a and filopodia distribution in African green monkey kidney COS-7 cells. High sensitivity TIRF technology and two channels imaging method provided a wide application in bio-medical studies. PMID:21137398

Liu, Xiao-chen; Guan, Li-zhao; Ma, Wan-yun; Zhang, Hong-quan

2010-10-01

209

Synthesis of Near-IR Fluorescent Oxazine Dyes with Esterase-labile Sulfonate Esters  

PubMed Central

Near-IR oxazine dyes are reported that contain sulfonate esters which are rapidly cleaved by esterase activity to unmask highly polar anionic sulfonates. Strategies for the synthesis of these dyes included the development of milder dye condensation conditions with improved functional compatibility, and the use of an alkyl halide that allows for the introduction of esterase-labile sulfonates without the need for sulfonation of the target molecule. PMID:22047733

Pauff, Steven M.

2011-01-01

210

Role of rare earth oxide nanoparticles (CeO2 and La2O3) in suppressing the photobleaching of fluorescent organic dyes.  

PubMed

Aqueous solutions with Rhodamine dye, and fluorescently labeled polymer samples of fibrin and collagen were mixed with aqueous dispersions of cerium oxide, lanthanum oxide, iron (II) oxide nanoparticles, and OxyFluor, a commonly used reagent for suppressing photobleaching. From time dependent studies of the fluorescence from these samples, we observed that the dyes in samples containing rare earth oxide nanoparticles exhibited significantly slower rates of fluorescence decay compared to control samples without additives, or containing OxyFluor or iron oxide nanoparticles. We posit that this may be related to the oxygen free radical scavenging properties of rare earth oxides. PMID:24706286

Guha, Anubhav; Basu, Anindita

2014-05-01

211

Alexa and Oregon Green dyes as fluorescence anisotropy probes for measuring protein-protein and protein-nucleic acid interactions.  

PubMed

The fluorescence properties of Alexa 488, Oregon Green 488, and Oregon Green 514 (Molecular Probes (Eugene, OR)) are compared when conjugated to biomolecules and as model compounds free in solution. We show that these relatively new, green fluorescence probes are excellent probes for investigation of the thermodynamics of protein-protein and protein-nucleic acid interactions by fluorescence anisotropy. Unlike fluorescein, the emission of these dyes has minimal pH dependence near neutrality and is significantly less susceptible to photobleaching. Steady-state and time-resolved fluorescence anisotropy data are compared for two interacting proteins of different size and for the association of a transcription factor with a DNA oligonucleotide containing a specific binding site. The temperature dependence of the fluorescence lifetimes of the probes is reported, and the effects of molecular size and probe motion on steady-state anisotropy data are discussed. The critical interplay among correlation time, fluorescence lifetime, and the observed steady-state anisotropy is evaluated. PMID:12234459

Rusinova, Elena; Tretyachenko-Ladokhina, Vira; Vele, Oana E; Senear, Donald F; Alexander Ross, J B

2002-09-01

212

Sensitive spectroscopic detection of large and denatured protein aggregates in solution by use of the fluorescent dye Nile red.  

PubMed

The fluorescent dye Nile red was used as a probe for the sensitive detection of large, denatured aggregates of the model protein beta-galactosidase (E. coli) in solution. Aggregates were formed by irreversible heat denaturation of beta-galactosidase below and above the protein's unfolding temperature of 57.4 degrees C, and the presence of aggregates in heated solutions was confirmed by static light scattering. Interaction of Nile red with beta-galactosidase aggregates led to a shift of the emission maximum (lambda (max)) from 660 to 611 nm, and to an increase of fluorescence intensity. Time-resolved fluorescence and fluorescence correlation spectroscopy (FCS) measurements showed that Nile red detected large aggregates with hydrodynamic radii around 130 nm. By steady-state fluorescence measurements, it was possible to detect 1 nM of denatured and aggregated beta-galactosidase in solution. The comparison with size exclusion chromatography (SEC) showed that native beta-galactosidase and small aggregates thereof had no substantial effect on the fluorescence of Nile red. Large aggregates were not detected by SEC, because they were excluded from the column. The results with beta-galactosidase demonstrate the potential of Nile red for developing complementary analytical methods that overcome the size limitations of SEC, and can detect the formation of large protein aggregates at early stages. PMID:17294134

Sutter, Marc; Oliveira, Sabrina; Sanders, Niek N; Lucas, Bart; van Hoek, Arie; Hink, Mark A; Visser, Antonie J W G; De Smedt, Stefaan C; Hennink, Wim E; Jiskoot, Wim

2007-03-01

213

Isolation of full-size mRNA from ethanol-fixed cells after cellular immunofluorescence staining and fluorescence-activated cell sorting (FACS)  

SciTech Connect

Preparation of intact, full-size RNA from tissues or cells requires stringent precautions against ubiquitous and rather stable RNases. Fluorescence-activated cell sorting (FACS) usually aims at the isolation of cells according to cell surface markers on living cells, from which RNA can be obtained by standard protocols. The separation of cells according to intracellular immunofluorescence markers, such as intranuclear, intracytoplasmic, or secreted molecules, requires permeation of the cell membrane for the staining antibodies, which is usually achieved by fixation. However, commonly used fixatives such as ethanol, methanol, or formaldehyde do not inactivate RNases completely, thereby hampering the analysis of complete RNA molecules from fixed cells. We report isolation of intact, full size RNA suitable for Northern blotting from cells that were fixed by 95% ethanol/5% acetic acid containing RNase inhibitors, stained intracellularly, and sorted by FACS. 21 refs., 2 figs., 1 tab.

Esser, C.; Kremer, J.; Hundeiker, C. [Univ. of Duesseldorf (Germany); Goettlinger, C.; Radbruch, A. [Univ. of Cologne, Koeln (Germany)

1995-12-01

214

Study of foxing stains on paper by chemical methods, infrared spectroscopy, micro-X-ray fluorescence spectrometry and laser induced breakdown spectroscopy  

NASA Astrophysics Data System (ADS)

Foxing spots appear on the paper as stains of reddish-brown, brown or yellowish color, generally of small dimensions, with sharp or irregular edges, most of which, if excited with UV light, show fluorescence. The formation mechanisms of foxed areas have been studied since 1935, however, despite more recent intensive research there are still no conclusive results. Some authors found evidence of bacterial or fungal growth in some foxed areas sometimes associated with the presence of iron. We decided to focus our attention on the influence of the different iron valence in the formation of stains in the paper. For this reason we artificially induced the formation of foxing by adding to the paper small, known quantities of iron (III) and iron (II) ions. We prepared aqueous solutions of ferric chloride and ferrous sulfate at three different concentrations and we always used the same quantity of each solution (5 ?l) to obtain a foxing stain. Part of the paper samples was artificially aged in a climatic chamber at 80 °C, 65% relative humidity for 15 days and part was submitted to aging for the same period at ambient temperatures under UV light at 240 nm. All papers were then analyzed for stain diameter, chromaticity coordinates, fluorescence under UV illumination, water content in the paper and in the spots, carbonyl content and then examined with infrared spectroscopy, X-ray fluorescence spectrometry and laser induced breakdown spectroscopy. Infrared spectra were collected in transmittance from potassium bromide pellets or directly in reflectance under microscope; X-ray fluorescence analysis were carried out using an X-ray microbeam (350 ?m beam spot; W X-ray tube) and LIBS analysis with Nd:YAG laser coupled with a Czerny-Turner spectrometer. As a result it is stated that the foxing phenomenon is related to a strong oxidation of the cellulose chain. Concerning the color coordinates there are no great differences between samples treated with iron (III) and iron (II). Carbonyl content, on the contrary, varies for the two sets of samples, especially in relation with the kind of aging. ?-XRF and LIBS measurements show a relationship between iron valence and calcium ion displacement in the foxed areas.

Bicchieri, M.; Ronconi, S.; Romano, F. P.; Pappalardo, L.; Corsi, M.; Cristoforetti, G.; Legnaioli, S.; Palleschi, V.; Salvetti, A.; Tognoni, E.

2002-07-01

215

A universal and label-free aptasensor for fluorescent detection of ATP and thrombin based on SYBR Green I dye.  

PubMed

A facile and universal aptamer-based label-free approach for selective and sensitive fluorescence detection of proteins and small biomolecules by using the SYBR Green I (SGI) dye is developed. This robust versatile biosensing strategy relies on fluorescence turn-off changes of SGI, resulting from target-induced structure switching of aptamers. Upon binding with the targets, the aptamers dissociate from the respective cDNA/aptamer duplexes, leading to the release of the dsDNA-intercalated SGI into solution and the quenching of the corresponding fluorescence intensities. Such target-induced conformational changes and release of aptamers from the DNA duplexes essentially lead to the change in the fluorescence signal of the SGI and thus constitute the mechanism of our aptamer-based label-free fluorescence biosensor for specific target analyses. Under optimized conditions, our method exhibits high sensitivity and selectivity for the quantification of ATP and thrombin with low detection limits (23.4 nM and 1.1 nM, respectively). Compared with previous reported methods for aptamer-based detection of ATP and thrombin, this label-free approach is selective, simple, convenient and cost-efficient without any chemical labeling of the probe or the target. Therefore, the present strategy could be easily applicable to biosensors that target a wide range of biomolecules. PMID:23202351

Kong, Ling; Xu, Jin; Xu, Yunying; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

2013-04-15

216

Interactions of L-Arg with calf thymus DNA using neutral red dye as a fluorescence probe  

NASA Astrophysics Data System (ADS)

The interaction between L-Arg and calf thymus DNA (ctDNA) in sodium acetate-acetic acid buffer (pH = 4) was investigated with the use of neutral red (NR) dye as a spectral probe coupled with UV-vis absorption, fluorescence, and circular dichroism (CD) spectroscopy technique. The UV absorption spectroscopy indicated that L-Arg interacted with ctDNA via electrostatic force and the fluorescence enhancing of the DNA-NR system verified the electrostatic interaction. In addition, detectable changes in the CD spectrum of ctDNA in the presence of L-Arg indicated conformational changes in the DNA double helix after interaction with the drug. Docking studies were found to corroborate the experimental results. All these results prove that this drug interacts with ctDNA via an electrostatic binding mode.

Lin, Jing; Liu, Rutao; Gao, Canzhu

2012-11-01

217

A Long-Wavelength Fluorescent Squarylium Cyanine Dye Possessing Boronic Acid for Sensing Monosaccharides and Glycoproteins with High Enhancement in Aqueous Solution  

PubMed Central

Fluorescence sensing of saccharides and glycoproteins using a boronic acid functionalized squarylium cyanine dye (“SQ-BA”) is characterized in terms of synthetic, fluorometric, thermodynamic and kinetic parameters. In our previous work, this newly synthesized dye was successfully applied to the separation and quantification of Gram-positive bacteria by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF); however, the fundamental properties of the dye and its saccharide complexes still required elucidation, as presented in this paper. The dye itself forms nonemissive, soluble aggregates in aqueous solution. With the addition of a monosaccharide, the dye aggregate dissociates to form an emissive monomer accompanied by the formation of a cyclic cis-diol ester with long-wavelength emission (?ex = 630 nm, ?em = 660 nm). A very large fluorescence enhancement factor of 18× was observed for the sensing dye as a fructose complex at pH 10, yielding a limit of detection of 10 ?M fructose. The relative order of fluorescence enhancement of SQ-BA with other monosaccharides was found to be: fructose > ribose > arabinose ? galactose > xylose > mannose > rhamnose > fucose ? glucose; and apparent affinity constants of 102.80, 102.08 and 100.86 M?1 were determined for fructose, ribose and glucose, respectively. Formation of the emissive complexes occurred within minutes, proving the kinetics of the sugar-dye interactions to be suitable for on-column labeling methods in CE-LIF. Furthermore, the sensing dye was successfully applied to glycoproteins, mucin type I–S and type III, which were detected with high sensitivity in batch aqueous solution as a result of the sugar-selective boronic acid-diol esterification as well as hydrophobic interactions. PMID:22778592

Saito, Shingo; Massie, Tara L.; Maeda, Takeshi; Nakazumi, Hiroyuki; Colyer, Christa L.

2012-01-01

218

In-situ investigation of adsorption of dye and coadsorbates on TiO2 films using QCM-D, fluorescence and AFM techniques  

NASA Astrophysics Data System (ADS)

Simultaneous adsorption of dye molecules and coadsorbates is important for the fabrication of high-efficiency dyesensitized solar cells, but its mechanism is not well understood. Herein, we use a quartz crystal microbalance with dissipation technique (QCM-D) to study dynamically and quantitatively the sensitization of TiO2 in situ. We investigate dye loading for a ruthenium(II) polypyridyl complex (Z907), of a triphenylamine-based D-?-A dye (Y123), and of a ullazine sensitizer (JD21), as well as the simultaneous adsorption of the latter two with the coadsorbate chenodeoxycholic acid. By combining the QCM-D technique with fluorescence measurements, we quantify molar ratios between the dye and coadsorbate. Furthermore, we will present first studies using liquid-phase AFM on the adsorbed dye monolayer, thus obtaining complementary microscopic information that may lead to understanding of the adsorption mechanism on the molecular scale.

Harms, Hauke A.; Tétreault, Nicolas; Voitchovsky, Kislon; Stellacci, Francesco; Grätzel, Michael

2013-09-01

219

A highly sensitive fluorescent indicator dye for calcium imaging of neural activity in vitro and in vivo  

PubMed Central

Calcium imaging of individual neurons is widely used for monitoring their activity in vitro and in vivo. Synthetic fluorescent calcium indicator dyes are commonly used, but the resulting calcium signals sometimes suffer from a low signal-to-noise ratio (SNR). Therefore, it is difficult to detect signals caused by single action potentials (APs) particularly from neurons in vivo. Here we showed that a recently developed calcium indicator dye, Cal-520, is sufficiently sensitive to reliably detect single APs both in vitro and in vivo. In neocortical neurons, calcium signals were linearly correlated with the number of APs, and the SNR was > 6 for in vitro slice preparations and > 1.6 for in vivo anesthetised mice. In cerebellar Purkinje cells, dendritic calcium transients evoked by climbing fiber inputs were clearly observed in anesthetised mice with a high SNR and fast decay time. These characteristics of Cal-520 are a great advantage over those of Oregon Green BAPTA-1, the most commonly used calcium indicator dye, for monitoring the activity of individual neurons both in vitro and in vivo. PMID:24405482

Tada, Mayumi; Takeuchi, Atsuya; Hashizume, Miki; Kitamura, Kazuo; Kano, Masanobu

2014-01-01

220

Evaluation of quantum dot-based concentric FRET configurations with a fluorescent dye and dark quencher for multiplexed bioanalyses  

NASA Astrophysics Data System (ADS)

Semiconductor quantum dots (QDs) continue to emerge as a highly advantageous platform for bioanalysis. Their unique physical and optical properties are especially well suited for Förster resonance energy transfer (FRET)-based bioprobes. Concentric FRET configurations are a recent development in this area of research and are best described as QD bioconjugates where multiple energy transfer pathways have been assembled around the central QD. Concentric FRET configurations permit multiplexed bioanalysis using one type of QD vector, but require more sophisticated analyses than conventional FRET pairs. In this paper, we describe the design and characterization of a new concentric FRET configuration that assembles both a fluorescent dye, Alexa Fluor 555 or Alexa Fluor 647, and a dark quencher, QSY9, at different ratios around a central CdSeS/ZnS QD. It was found that the magnitudes of the total photoluminescence (PL) intensity and either the A555/QD or A647/QD PL ratio can be related to the number of QSY9 and A555 or A647 per QD. The trends in these parameters with changes in the number of each dye molecule per QD have both similarities and differences between configurations with A555 and A647. In each case, a system of equations can be defined to permit calculation of the number of each dye molecule per QD from PL measurements. Both of these dark quencher-based concentric FRET configurations are therefore good candidates for quantitative, multiplexed bioanalysis.

Conroy, Erin M.; Algar, W. Russ

2014-03-01

221

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Glazer, Alexander M. (Orinde, CA); Benson, Scott C. (Albany, CA)

1998-01-01

222

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Glazer, Alexander M. (Orinda, CA); Benson, Scott C. (Albany, CA)

1999-01-01

223

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Glazer, Alexander N. (Orinda, CA); Benson, Scott C. (Albany, CA)

1997-01-01

224

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figures.

Glazer, A.N.; Benson, S.C.

1995-03-28

225

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figs.

Glazer, A.M.; Benson, S.C.

1998-06-16

226

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated. 4 figs.

Glazer, A.N.; Benson, S.C.

1997-07-08

227

DNA complexes with dyes designed for energy transfer as fluorescent markers  

DOEpatents

Heteromultimeric fluorophores are provided for binding to DNA, which allow for the detection of DNA in electrical separations and preparation of probes having high-fluorescent efficiencies and large Stokes shifts. In addition, by appropriate choice of fluorescent molecules, one can use a single narrow wavelength band excitation light source, while obtaining fluorescent emissions having sufficient separation to be readily discriminated.

Glazer, Alexander N. (Orinda, CA); Benson, Scott C. (Albany, CA)

1995-01-01

228

Calmodulin activation of the Ca2+ pump revealed by fluorescent chelator dyes in human red blood cell ghosts  

PubMed Central

Ca2+ transport in red blood cell ghosts was monitored with fura2 or quin2 incorporated as the free acid during resealing. This is the first report of active transport monitored by the fluorescent intensity of the chelator dyes fura2 (5-50 microM) or quin2 (250 microM) in hemoglobin-depleted ghosts. Since there are no intracellular compartments in ghosts and the intracellular concentrations of all assay chelator substances including calmodulin (CaM), the dyes, and ATP could be set, the intracellular concentrations of free and total Ca [( Cafree]i and [Catotal]i) could be calculated during the transport. Ghosts prepared with or without CaM rapidly extruded Ca2+ to a steady- state concentration of 60-100 nM. A 10(4)-fold gradient for Ca2+ was routinely produced in medium containing 1 mM Ca2+. During active Ca2+ extrusion, d[Cafree]i/dt was a second order function of [Cafree]i and was independent of the dye concentration, whereas d[Catotal]i/dt increased as a first order function of both the [Cafree]i and the concentration of the Ca:dye complex. CaM (5 microM) increased d[Catotal]i/dt by 400% at 1 microM [Cafree]i, while d[Cafree]i/dt increased by only 25%. From a series of experiments we conclude that chelated forms of Ca2+ serve as substrates for the pump under permissive control of the [Cafree]i, and this dual effect may explain cooperativity. Free Ca2+ is extruded, and probably also Ca2+ bound to CaM or other chelators, while CaM and the chelators are retained in the cell. PMID:1371307

1992-01-01

229

Measurement of Bluetongue Virus Binding to a Mammalian Cell Surface Receptor by an In Situ Immune Fluorescent Staining Technique  

Technology Transfer Automated Retrieval System (TEKTRAN)

A quantifiable in situ immune fluorescent assay (IFA) was developed to measure bluetongue virus (BTV) binding to mammalian cells. The utility of the assay was demonstrated with both Chinese hamster ovary (CHO) and bovine pulmonary artery endothelial (CPAE) cells. Since heparin sulfate (HS) has been ...

230

Time-Saving Benefits of Intravital Staining.  

PubMed

One of the challenges in labeling tissues for fluorescence microscopy is minimizing sample processing while maintaining or improving the information generated by the fluorescent label. Generally, tissues are extracted, fixed, and embedded in mounting media (such as paraffin), sectioned, and then postprocessed by removing the paraffin, blocking, labeling, and washing. Despite all of these steps, the consistency of labeling quality can vary as a result of several factors, including heterogeneity in labeling efficiency from slide to slide, the necessity of postprocessing to obtain information on sequential sections of tissue, interference from the mounting media, and loss of native three-dimensional structural information, especially in thicker sections. A method for embedding and processing tissues that have been labeled by intravital staining is described in this study. Intravital staining is the process in which live-cell dyes and other labels are injected into the bloodstream before fixation of the tissues. Tissues processed this way can be imaged upon sectioning without further staining and retain their native, three-dimensional information, thereby improving the information retained by the labels and speeding up sample processing. PMID:20622939

Macgillivray, Catherine; Sylvan, Jeremy; Lee, Richard T; Huang, Hayden

2008-09-01

231

Dye Like A Natural  

NSDL National Science Digital Library

In this activity, learners stain fabrics--on purpose! Learners explore the art of natural dyeing by using dyes and substrates that are both derived from plant or animal sources as well as mordant solutions. Learners compare the color and effectiveness of different mordant/dye combinations on the different substrates.

Yu, Julie

2010-01-01

232

Investigation of time-resolved fluorescence lifetime of perylene dye molecules embedded in silicon nanopillars  

NASA Astrophysics Data System (ADS)

The radiative decay rate of a perylene dye molecule attached to silicon nanopillar is investigated using a conventional time-correlated single photon counting technique. It is hard to produce a sustainable host with exactly the same dimensions all the time during fabrication to accommodate dye molecules for enhancement of spontaneous emission rate. The laser-induced electrochemical anodization method allows us to have a control over size and shape of the silicon nanostructures. The effect of the silicon nanopillar on the radiative decay rate of the dye molecules is described by the Klimov's prolate nanospheroid model. It is observed that the decay rate is significantly enhanced or inhibited due to plasmon resonance, depending on whether the dipole is embedded closely right at the tip or at equator of the prolate nanospheroid. Both inhibition and enhancement disappear when the distance between the dipole and prolate nanospheroid becomes large. Thus, the decay rate of the dye molecule approaches its natural value in the free space.

Acikgoz, Sabriye

2015-02-01

233

Bacterial population dynamics in a reverse-osmosis water purification system determined by fluorescent staining and PCR-denaturing gradient gel electrophoresis.  

PubMed

The bacterial population dynamics in an industrial scale reverse-osmosis (RO) water purification system were analyzed by fluorescent staining methods and denaturing gradient gel electrophoresis (DGGE). Bacterial numbers increased with storage in a tank, and bacterial diversity changed during the water purification process. A DNA sequence-based analysis of the major bands on the DGGE gel revealed that Simonsiella sp. (Betaproteobacteria) was abundant in the source water (activated sludge-treated waste effluent), while Bosea sp. and Rhizobium sp. (Alphaproteobacteria), which usually exist in an oligotrophic environment, became abundant during the water purification process. These results suggest the importance of microbiological monitoring by culture-independent methods for quality control in RO water purification systems. These methods could provide an early warning of impending problems and clarify critical steps in controlling specific bacteria contributing to the contamination of RO water systems. PMID:21566369

Baba, Takashi; Matsumoto, Rie; Yamaguchi, Nobuyasu; Nasu, Masao

2009-01-01

234

Ultrasound-guided fine-needle aspiration of a posterior neck dedifferentiated liposarcoma with MDM2 fluorescence in situ hybridization performed on a Pap-stained smear.  

PubMed

Head and neck liposarcomas, while rare, tend to be subcutaneous and well-differentiated. Dedifferentiated liposarcomas of the head and neck are exceedingly rare in the literature. We present a case of a dedifferentiated liposarcoma arising in the soft tissue of the posterior neck of an 86-year-old man and diagnosed by fine-needle aspiration. Aspirate smears showed a dual population of atypical lipomatous and spindled cells. MDM2 (murine double minute 2) amplification was demonstrated on a Pap-stained smear using fluorescence in situ hybridization (FISH). To the best of our knowledge, this is the first report of MDM2 FISH amplification in a liposarcoma performed on an aspirate smear. Diagn. Cytopathol. 2014. © 2014 Wiley Periodicals, Inc. PMID:25132684

Zreik, Riyam; Soyalp, Krystal; Ruiz, Steve; Ward, Russell; Dobin, Sheila; Chen, Xiangbai; Liu, Lina; Rao, Arundhati

2014-07-31

235

Application of a vital fluorescent staining method for simultaneous, near-real-time concentration monitoring of two bacterial strains in an Atlantic coastal plain aquifer in Oyster, Virginia.  

PubMed

Two differentially labeled bacterial strains were monitored in near-real time during two field-scale bacterial transport experiments in a shallow aquifer in July 2000 and July 2001. Comamonas sp. strain DA001 and Acidovorax sp. strain OY-107 were grown and labeled with the vital fluorescent stain TAMRA/SE (5 [and -6]-carboxytetramethylrhodamine, succinimidyl ester) or CFDA/SE (5 [and -6]-carboxyfluorescein diacetate, succinimidyl ester). Fluorescently labeled cells and a conservative bromide tracer were introduced into a suboxic superficial aquifer, followed by groundwater collection from down-gradient multilevel samplers. Cells were enumerated in the field by microplate spectrofluorometry, with confirmatory analyses for selected samples done in the laboratory by epifluorescence microscopy, flow cytometry, and ferrographic capture. There was general agreement in the results from all of the vital-stain-based enumeration methods, with differences ranging from <10% up to 40% for the analysis of identical samples between different tracking methods. Field analysis by microplate spectrofluorometry was robust and efficient, allowing thousands of samples to be analyzed in quadruplicate for both of the injected strains. The near-real-time data acquisition allowed adjustments to the predetermined sampling schedule to be made. The microplate spectrofluorometry data sets for the July 2000 and July 2001 experiments allowed the transport of the injected cells to be related to the site hydrogeology and injection conditions and enabled the assessment of differences in the transport of the two strains. This near-real-time method should prove effective for a number of microbial ecology applications. PMID:15006793

Fuller, Mark E; Mailloux, Brian J; Streger, Sheryl H; Hall, James A; Zhang, Pengfei; Kovacik, William P; Vainberg, Simon; Johnson, William P; Onstott, Tullis C; DeFlaun, Mary F

2004-03-01

236

Application of a Vital Fluorescent Staining Method for Simultaneous, Near-Real-Time Concentration Monitoring of Two Bacterial Strains in an Atlantic Coastal Plain Aquifer in Oyster, Virginia  

PubMed Central

Two differentially labeled bacterial strains were monitored in near-real time during two field-scale bacterial transport experiments in a shallow aquifer in July 2000 and July 2001. Comamonas sp. strain DA001 and Acidovorax sp. strain OY-107 were grown and labeled with the vital fluorescent stain TAMRA/SE (5 [and -6]-carboxytetramethylrhodamine, succinimidyl ester) or CFDA/SE (5 [and -6]-carboxyfluorescein diacetate, succinimidyl ester). Fluorescently labeled cells and a conservative bromide tracer were introduced into a suboxic superficial aquifer, followed by groundwater collection from down-gradient multilevel samplers. Cells were enumerated in the field by microplate spectrofluorometry, with confirmatory analyses for selected samples done in the laboratory by epifluorescence microscopy, flow cytometry, and ferrographic capture. There was general agreement in the results from all of the vital-stain-based enumeration methods, with differences ranging from <10% up to 40% for the analysis of identical samples between different tracking methods. Field analysis by microplate spectrofluorometry was robust and efficient, allowing thousands of samples to be analyzed in quadruplicate for both of the injected strains. The near-real-time data acquisition allowed adjustments to the predetermined sampling schedule to be made. The microplate spectrofluorometry data sets for the July 2000 and July 2001 experiments allowed the transport of the injected cells to be related to the site hydrogeology and injection conditions and enabled the assessment of differences in the transport of the two strains. This near-real-time method should prove effective for a number of microbial ecology applications. PMID:15006793

Fuller, Mark E.; Mailloux, Brian J.; Streger, Sheryl H.; Hall, James A.; Zhang, Pengfei; Kovacik, William P.; Vainberg, Simon; Johnson, William P.; Onstott, Tullis C.; DeFlaun, Mary F.

2004-01-01

237

Fluorescence-based, high-throughput assays for ?-opioid receptor activation using a membrane potential-sensitive dye.  

PubMed

The development of new and improved opioid analgesics requires high-throughput screening (HTS) methods to identify potential therapeutics from large libraries of lead compounds. Here we describe two simple, real-time fluorescence-based assays of ?-opioid receptor activation that may be scaled up for HTS. In AtT-20 cells expressing the ?-opioid receptor (MOPr), opioids activate endogenous G protein gated inwardly rectifying K channels (GIRK channels), leading to membrane hyperpolarization. In Chinese hamster ovary cells expressing MOPr, adenylyl cyclase activation via forskolin results in membrane hyperpolarization, which is inhibited by opioids. Changes in membrane potential can be measured using a proprietary membrane potential-sensitive dye. In contrast to many HTS methods currently available, these assays reflect naturalistic coupling of the receptor to effector molecules. PMID:25293325

Knapman, Alisa; Connor, Mark

2015-01-01

238

Dual color fluorescence quantitative detection of specific single-stranded DNA with molecular beacons and nucleic acid dye SYBR Green I.  

PubMed

We have developed a dual color fluorescence quantitative detection method for specific single-stranded DNA with molecular beacons (MBs) and nucleic acid dye SYBR Green I by synchronous scanning fluorescence spectrometry. It is demonstrated by a reverse-transcription oligonucleotide sequence (target DNA, 33 bases) of RNA fragment of human immunodeficiency virus (HIV) as a model system. In the absence of target DNA, the MBs are in the stem-closed state, the fluorescence of 5-carboxy-X-rhodamine (ROX) is quenched by black hole quencher-2 (BHQ-2), and the interaction between SYBR Green I and the MBs is very weak. At this time the fluorescence signals of ROX and SYBR Green I are all very weak. In the presence of target DNA, MBs hybridize with target DNA and form a double-strand structure, the fluorophore ROX is separated from the quencher BHQ-2, and the fluorescence of ROX recovers. At the same time, SYBR Green I binds to hybridized dsDNA, whose fluorescence intensity is significantly enhanced. Thus, dual color fluorescence quantitative detection for the target DNA can be realized by synchronous scanning fluorescence spectrometry. In this strategy, the fluorescence signal of SYBR Green I is far larger than that of ROX, so the quantitative analysis of target DNA with the fluorescence intensity of SYBR Green I can significantly improve the detection sensitivity. In addition, the false-positive signals of MBs do not affect the fluorescence signals of nucleic acid dye SYBR Green I. Thereby, in the analysis of complex samples, quantitative analysis of target DNA with SYBR Green I can avoid the false-positive signals of MBs and improve the detection accuracy. PMID:22763945

Xiang, Dong-Shan; Zhou, Guo-Hua; Luo, Ming; Ji, Xing-Hu; He, Zhi-Ke

2012-08-21

239

A robust and versatile signal-on fluorescence sensing strategy based on SYBR Green I dye and graphene oxide  

PubMed Central

A robust and versatile signal-on fluorescence sensing strategy was developed to provide label-free detection of various target analytes. The strategy used SYBR Green I dye and graphene oxide as signal reporter and signal-to-background ratio enhancer, respectively. Multidrug resistance protein 1 (MDR1) gene and mercury ion (Hg2+) were selected as target analytes to investigate the generality of the method. The linear relationship and specificity of the detections showed that the sensitive and selective analyses of target analytes could be achieved by the proposed strategy with low detection limits of 0.5 and 2.2 nM for MDR1 gene and Hg2+, respectively. Moreover, the strategy was used to detect real samples. Analytical results of MDR1 gene in the serum indicated that the developed method is a promising alternative approach for real applications in complex systems. Furthermore, the recovery of the proposed method for Hg2+ detection was acceptable. Thus, the developed label-free signal-on fluorescence sensing strategy exhibited excellent universality, sensitivity, and handling convenience. PMID:25565810

Qiu, Huazhang; Wu, Namei; Zheng, Yanjie; Chen, Min; Weng, Shaohuang; Chen, Yuanzhong; Lin, Xinhua

2015-01-01

240

Diversity-Oriented Facile Access to Highly Fluorescent Membrane-Permeable Benz[c,d]indole N-Heteroarene BF2 Dyes.  

PubMed

A diversity-oriented one-pot synthesis of a series of membrane-permeable BF2-rigidified benz[c,d]indole N-heteroarene BBN and BBC dyes has been achieved from the condensation of two commercial components (benz[c,d]indol-2-one and a set of N-heteroarene derivatives that can be selected from thousands of commercially available sources) and the subsequent in situ BF2 complexation reaction. These dyes enjoy a set of excellent photophysical properties including the large Stokes shift, high solution and solid-state fluorescence, and excellent photostabilities. PMID:25551331

Cheng, Chi; Gao, Naixun; Yu, Changjiang; Wang, Zhaoyun; Wang, Jun; Hao, Erhong; Wei, Yun; Mu, Xiaolong; Tian, Yanli; Ran, Chongzhao; Jiao, Lijuan

2015-01-16

241

Fluorescent-dye-doped sol-gel sensor for highly sensitive carbon dioxide gas detection below atmospheric concentrations.  

PubMed

Optical fluorescence sol-gel sensors have been developed for the detection of carbon dioxide gas in the 0.03-30% range with a detection limit of 0.008% (or 80 ppm) and a quantitation limit of 0.02% (or 200 ppm) CO(2). Sol-gels were spin-coated on glass slides to create an organically modified silica-doped matrix with the 1-hydroxypyrene-3,6,8-trisulfonate (HPTS) fluorescent indicator. The luminescence intensity of the HPTS indicator (513 nm) is quenched by CO(2), which protonates the anionic form of HPTS. An ion pair technique was used to incorporate the lipophilic dye into the hydrophilic sol-gel matrix. TiO(2) particles (<5 microm diameter) were added to induce Mie scattering and increase the incident light interaction with the sensing film, thus increasing the signal-to-noise ratio. Moisture-proof overcoatings have been used to maintain a constant level of water inside the sensor films. The optical sensors are inexpensive to prepare and can be easily coupled to fiber optics for remote sensing capabilities. A fiber-optic bundle was used for the gas detection and shown to work as part of a multianalyte platform for simultaneous detection of multiple analytes. The studies reported here resulted in the development of sol-gel optical fluorescent sensors for CO(2) gas with sensitivity below that in the atmosphere (ca. 387 ppm). These sensors are a complementary approach to current FT-IR measurements for real-time carbon dioxide detection in environmental applications. PMID:20038093

Dansby-Sparks, Royce N; Jin, Jun; Mechery, Shelly J; Sampathkumaran, Uma; Owen, Thomas William; Yu, Bi Dan; Goswami, Kisholoy; Hong, Kunlun; Grant, Joseph; Xue, Zi-Ling

2010-01-15

242

Adsorption of fluorescent R6G dye into organophilic C12TMA laponite films  

Microsoft Academic Search

The absorption and fluorescence properties of rhodamine 6G (R6G) in organophilic laponite (Lap) clay films are studied. For this purpose, organo-Lap clays are synthesized by the incorporation of dodecyltrimethylammonium (C12TMA) as surfactant into the interlayer space of Lap clays. Two organo-Lap clays are prepared: one with moderate surfactant content (around 70% of the total cation-exchange capacity (CEC) of the clay)

S. Salleres; F. López Arbeloa; V. Martínez; T. Arbeloa; I. López Arbeloa

2008-01-01

243

High pressure study of flexible fluorescent dye molecules in solid polymeric media. I. Julolidinemalononitrile  

NASA Astrophysics Data System (ADS)

The pressure effect on the fluorescence of julolidinemalononitrile (JDMN) in several solid polymers has been studied. An increase of fluorescence intensity by a factor of 8 to 15 has been observed, in different polymers, within 120 kbar. Two general regions of pressure behavior have been distinguished, in all polymers. In the first pressure region (0-20 kbar), the amplification of fluorescence intensity is explained by a retardation of motion (twist) in the excited state by increasing pressure. This process is shown to be controlled by an energy barrier between two conformations of molecule (planar "P" and twisted "T") in its excited state. The relaxation from one form to the other is a single process reflecting thus one possible twist of the malonitrile group. The exponential relation obtained between nonradiative rate for intramolecular twist ( k' nr) and emitting state energy ( EfP) implies the pressure insensitivity of the energy between intersection of the potential wells for "P" and "T" excited states relative to the "P" ground state. In the second pressure region (>40 kbar), the intensity decrease is controlled by the nonradiative rate knrp for the originally excited state. It has been shown that the energy gap law is applicable to explain experimental data in this region.

Dreger, Z. A.; Lang, J. M.; Drickamer, H. G.

1993-02-01

244

Vacuolar staining methods in plant cells.  

PubMed

Commercially available fluorescent dyes enable the fast and specific visualization of plant vacuoles, allowing for investigation of membrane dynamics and vacuolar biogenesis in living cells. Here, we describe different approaches tinting the tonoplast or the vacuolar lumen with a range of dyes, and illustrate its utilization with established fluorescent-tagged marker lines. PMID:25408446

Scheuring, David; Schöller, Maria; Kleine-Vehn, Jürgen; Löfke, Christian

2015-01-01

245

Label-free and sensitive fluorescence detection of nucleic acid, based on combination of a graphene oxid /SYBR green I dye platform and polymerase assisted signal amplification  

NASA Astrophysics Data System (ADS)

A new label-free isothermal fluorescence amplification detection for nucleic acid has been developed. In this paper, we first developed a novel sensitive and specific detection platform with an unmodified hairpin probe (HP) combination of the graphene oxid (GO)/ SYBR green I dye (SG), which was relied on the selective principle of adsorption and the high quenching efficiency of GO. Then for the application of this new strategy, we used Mirco RNA-21 (Mir-21) as the target to evaluate this working principle of our design. When the target was hybridizing with the HP and inducing its conformation of change, an efficient isothermal circular strand-displacement polymerization reaction was activating to assist the first signal amplification. In this format, the formed complex conformation of DNA would interact with its high affinity dye, then detached from the surface of GO after incubating with the platform of GO/intercalating dye. This reaction would accompany with obvious fluorescence recovery, and accomplish farther signal enhancement by a mass of intercalating dye inserting into the minor groove of the long duplex replication product. By taking advantage of the multiple amplification of signal, this method exerted substantial enhancement in sensitivity and could be used for rapid and selective detection of Mir-21 with attomole range. It is expected that this cost-effective GO based sensor might hold considerable potential to apply in bioanalysis studies.

Zhu, Xiao; Xing, Da

2012-12-01

246

Integrating fluorescent dye flow-curve testing and acoustic Doppler velocimetry profiling for in situ hydraulic evaluation and improvement of clarifier performance.  

PubMed

Enhancing the performance of clarifiers requires a thorough understanding of their hydraulics. Fluorescence spectroscopy and acoustic doppler velocimeter (ADV) profiling generally have been used separately to evaluate secondary settlers. We propose that simultaneous use of these techniques is needed to obtain a more reliable and useful evaluation. Experiments were performed on laboratory- and full-scale clarifiers. Factors affecting Fluorescein and Rhodamine 6G properties were identified. Underestimations up to 500% in fluorescence intensities may be derived from differential fluorescence quenching by oxygen. A careful control and interpretation of fluorescent dye experiments is needed to minimize artifacts in real settings. While flow-curve tests constructed under controlled conditions provided a more accurate overall quantitative estimation of the hydraulic performance, ADV velocity and turbulence profiling provided a detailed spatial understanding of flow patterns that was used to troubleshoot and fix the causes of hydraulic short-circuits. PMID:20853746

Tarud, F; Aybar, M; Pizarro, G; Cienfuegos, R; Pastén, P

2010-08-01

247

Method for measuring the three-dimensional distribution of a fluorescent dye in a cell membrane  

NASA Astrophysics Data System (ADS)

This letter reports on a method for accurately determining the component distribution in a cell membrane over the entire cell surface. This method involves exciting a fluorescent-dyed cell membrane using evanescent light and scanning the entire cell surface by rotating the cell using a noncontact technique, namely, proximal two-beam optical tweezers. To position the cell membrane in the thin evanescent field, the authors designed an optical system capable of precisely positioning the focal position. Using this method, they were able to measure the surface distribution of glycoprotein labeled by lectin in a breast cancer cell membrane.

Yamamoto, Kazuya; Ishimaru, Ichirou; Fujii, Yoshiki; Yasokawa, Toshiki; Kuriyama, Shigeki; Masaki, Tsutomu; Takegawa, Kaoru; Tanaka, Naotaka

2007-01-01

248

Fluorescence Ratiometric Properties Induced by Nanoparticle Plasmonics and Nanoscale Dye Dynamics  

PubMed Central

Nanoscale transport of merocyanine 540 within/near the plasmon field of gold nanoparticles was recognized as an effective inducer of single-excitation dual-emission ratiometric properties. With a high concentration of the signal transducer (ammonium), a 700% increase in fluorescence was observed at the new red-shifted emission maximum, compared to a nanoparticle free sensor membrane. A previously nonrecognized isosbestic point is demonstrated at 581.4 ± 0.1?nm. The mechanism can be utilized for enhanced and simplified ratiometric optical chemical sensors and potentially for thin film engineering to make solar cells more effective and stable by a broader and more regulated absorption. PMID:23781159

2013-01-01

249

A SENSITIVE FLUORIMETRIC METHOD FOR THE DETERMINATION OF NITRITE AND NITRATE IN SEAWATER BY A NOVEL RED-REGION FLUORESCENCE DYE  

Microsoft Academic Search

A novel method for the determination of dissolved inorganic nitrite and nitrate in surface seawater was reported, which is based on the diazotization reaction between nitrite and a novel red-region fluorescent dye, tetra-substituted amino aluminum phthalocyanine (TAAlPc). Nitrate is determined as nitrite after reduction on a cadmium column. Under optimal conditions, the linear range of the calibration curve is 21–840

Xin-Qi Zhan; Dong-Hui Li; Hong Zheng; Jin-Gou Xu

2001-01-01

250

Retrograde labeling of retinal ganglion cells in adult zebrafish with fluorescent dyes.  

PubMed

As retrograde labeling retinal ganglion cells (RGCs) can isolate RGCs somata from dying sites, it has become the gold standard for counting RGCs in RGCs survival and regeneration experiments. Many studies have been performed in mammalian animals to research RGCs survival after optic nerve injury. However, retrograde labeling of RGCs in adult zebrafish has not yet been reported, though some alternative methods can count cell numbers in retinal ganglion cell layers (RGCL). Considering the small size of the adult zebrafish skull and the high risk of death after drilling on the skull, we open the skull with the help of acid-etching and seal the hole with a light curing bond, which could significantly improve the survival rate. After absorbing the dyes for 5 days, almost all the RGCs are labeled. As this method does not need to transect the optic nerve, it is irreplaceable in the research of RGCs survival after optic nerve crush in adult zebrafish. Here, we introduce this method step by step and provide representative results. PMID:24837333

Zou, Su-Qi; Tian, Chen; Du, Su-Tie; Hu, Bing

2014-01-01

251

Comparison of a chimeric anti-carcinoembryonic antigen antibody conjugated with visible or near-infrared fluorescent dyes for imaging pancreatic cancer in orthotopic nude mouse models  

NASA Astrophysics Data System (ADS)

The aim of this study was to evaluate a set of visible and near-infrared dyes conjugated to a tumor-specific chimeric antibody for high-resolution tumor imaging in orthotopic models of pancreatic cancer. BxPC-3 human pancreatic cancer was orthotopically implanted into pancreata of nude mice. Mice received a single intravenous injection of a chimeric anti-carcinoembryonic antigen antibody conjugated to one of the following fluorophores: 488-nm group (Alexa Fluor 488 or DyLight 488); 550-nm group (Alexa Fluor 555 or DyLight 550); 650-nm group (Alexa Fluor 660 or DyLight 650), or the 750-nm group (Alexa Fluor 750 or DyLight 755). After 24 h, the Olympus OV100 small-animal imaging system was used for noninvasive and intravital fluorescence imaging of mice. Dyes were compared with respect to depth of imaging, resolution, tumor-to-background ratio (TBR), photobleaching, and hemoglobin quenching. The longer wavelength dyes had increased depth of penetration and ability to detect the smallest tumor deposits and provided the highest TBRs, resistance to hemoglobin quenching, and specificity. The shorter wavelength dyes were more photostable. This study showed unique advantages of each dye for specific cancer imaging in a clinically relevant orthotopic model.

Maawy, Ali A.; Hiroshima, Yukihiko; Kaushal, Sharmeela; Luiken, George A.; Hoffman, Robert M.; Bouvet, Michael

2013-12-01

252

Modular video endoscopy for in vivo cross-polarized and vital-dye fluorescence imaging of Barrett's-associated neoplasia  

NASA Astrophysics Data System (ADS)

A modular video endoscope is developed and tested to allow imaging in different modalities. This system incorporates white light imaging (WLI), cross-polarized imaging (CPI), and vital-dye fluorescence imaging (VFI), using interchangeable filter modules. CPI and VFI are novel endoscopic modalities that probe mucosal features associated with Barrett's neoplasia. CPI enhances vasculature, while VFI enhances glandular architecture. In this pilot study, we demonstrate the integration of these modalities by imaging areas of Barrett's metaplasia and neoplasia in an esophagectomy specimen. We verify that those key image features are also observed during an in vivo surveillance procedure. CPI images demonstrate improved visualization of branching blood vessels associated with neoplasia. VFI images show glandular architecture with increased glandular effacement associated with neoplasia. Results suggests that important pathologic features seen in CPI and VFI are not visible during standard endoscopic white light imaging, and thus the modalities may be useful in future in vivo studies for discriminating neoplasia from Barrett's metaplasia. We further demonstrate that the integrated WLI/CPI/VFI endoscope is compatible with complementary high-resolution endomicroscopy techniques such as the high-resolution microendoscope, potentially enabling two-step ("red-flag" widefield plus confirmatory high-resolution imaging) protocols to be enhanced.

Thekkek, Nadhi; Pierce, Mark C.; Lee, Michelle H.; Polydorides, Alexandros D.; Flores, Raja M.; Anandasabapathy, Sharmila; Richards-Kortum, Rebecca R.

2013-02-01

253

Sputum direct fluorescent antibody (DFA)  

MedlinePLUS

Direct immunofluorescence test; Direct fluorescent antibody - sputum ... fluorescent dye are added to the sample. These antibodies are ... bright glow (fluorescence) can be seen in the sputum sample using ...

254

The use of vitamins as tracer dyes for laser-induced fluorescence in liquid flow applications  

NASA Astrophysics Data System (ADS)

Tracers commonly used in experimental flow studies are mostly nocuous to the environment and human health. Particularly, in large flow installations, this can become a problem. In this study, a solution of this problem is presented, based on using water-soluble vitamins. Five of them are examined here for their applicability in flow studies. Vitamins B2 and B6 turned out to be the most promising candidates, and the dependency of their fluorescence intensity on parameters like concentration, laser energy, temperature, and pH are determined for two commonly used laser excitation wavelengths (532, 355 nm). Two examples of application in a static mixer and a spray flow are shown and demonstrate the applicability of the vitamin tracers.

Zähringer, Katharina

2014-04-01

255

Papanicolaou staining--a review.  

PubMed

Some technical aspects of Papanicolaou staining are reviewed. The history of the technique is traced from Mallory's aniline blue technique through Masson's trichrome procedure to the techniques of Shorr. The histochemistry of the three Papanicolaou staining solutions (aluminum-"hematoxylin", OG and EA) is discussed. The structure of the aluminum-hematein chelate and its mode of action are considered. A tentative mechanism is proposed for the characteristic differential counterstaining produced by Papanicolaou techniques, i.e. orangeophilia vs cyanophilia. It is suggested that differential counterstaining occurs as a consequence of 1) differences in cytoplasmic density, 2) differences in the molecular size of the anionic dyes, and 3) the inhibitory effects of phosphotungstic acid on the binding of small dyes. The review considers some recent quantitative studies of Papanicolaou stained cells and outlines some modifications of Papanicolaou's procedures. The text concludes with a discussion of alternatives to the Papanicolaou technique. PMID:6195507

Marshall, P N

1983-05-01

256

application note Fluorescent protein staining  

E-print Network

Protein separation by 1-D or 2-D gel electrophoresis is one of the most widely used forms of protein For 1-D SDS polyacrylamide gels, low-molecular weight protein standards were separated on 15 of concentrated ammonia (sp. gr. 0.88­0.89). Gels were then left in signal stabilization solution (0.75% glacial

Lebendiker, Mario

257

Efficiency of staining hair with indocyanine green  

NASA Astrophysics Data System (ADS)

The efficiency of staining hair with indocyanine green (ICG) solution depending on type of hair, natural color, staining time and other parameters was investigated. Bonding ICG with hair material occurs due to interaction between ICG molecules and keratinocyte albumin. The penetration of ICG dye into hair meets with difficulties owing to surface protective layer.

Kulyabina, Tatyana V.; Kochubey, Vyacheslav I.

2005-06-01

258

Synthesis, spectroscopic and DFT studies of novel fluorescent dyes: 3-Aminoimidazo[1,2-a]pyridines possessing 4-pyrone moieties  

NASA Astrophysics Data System (ADS)

A series of novel imidazo[1,2-a]pyridines possessing 4-pyrone ring were synthesized by three-component condensation of 4-pyrone carbaldehydes, 2-aminopyridines and isocyanides. Bismuth (III) chloride was used as a catalyst in these reactions and desired products were synthesized in good yields at a very short period of time under solvent free conditions. UV-Vis absorption and fluorescence emission spectra of these compounds were investigated. It shown that two of these compounds (10f and 10g) exhibit intense fluorescence in dichloromethane. Optimized ground-state molecular geometries and orbital distributions of these two fluorescent dyes were obtained using density functional theory (DFT). Thermogravimetric analysis and electrochemical properties of these compounds were also studied.

Shahrisa, Aziz; Safa, Kazem Dindar; Esmati, Somayeh

2014-01-01

259

White emitters by tuning the excited-state intramolecular proton-transfer fluorescence emission in 2-(2'-hydroxybenzofuran)benzoxazole dyes.  

PubMed

The synthesis, structural, and photophysical properties of a new series of original dyes based on 2-(2'-hydroxybenzofuran)benzoxazole (HBBO) is reported. Upon photoexcitation, these dyes exhibit intense dual fluorescence with contribution from the enol (E*) and the keto (K*) emission, with K* being formed through excited-state intramolecular proton transfer (ESIPT). We show that the ratio of emission intensity E*/K* can be fine-tuned by judiciously decorating the molecular core with electron-donating or -attracting substituents. Push-pull dyes 9 and 10 functionalized by a strong donor (nNBu2 ) and a strong acceptor group (CF3 and CN, respectively) exhibit intense dual emission, particularly in apolar solvents such as cyclohexane in which the maximum wavelength of the two bands is the more strongly separated. Moreover, all dyes exhibit strong solid-state dual emission in a KBr matrix and polymer films with enhanced quantum yields reaching up to 54?%. A wise selection of substituents led to white emission both in solution and in the solid state. Finally, these experimental results were analyzed by time-dependent density functional theory (TD-DFT) calculations, which confirm that, on the one hand, only E* and K* emission are present (no rotamer) and, on the other hand, the relative free energies of the two tautomers in the excited state guide the ratio of the E*/K* emission intensities. PMID:25145709

Benelhadj, Karima; Muzuzu, Wenziz; Massue, Julien; Retailleau, Pascal; Charaf-Eddin, Azzam; Laurent, Adèle D; Jacquemin, Denis; Ulrich, Gilles; Ziessel, Raymond

2014-09-26

260

Long-term monitoring of live cell proliferation in presence of PVP-Hypericin: a new strategy using ms pulses of LED and the fluorescent dye CFSE  

PubMed Central

Summary During fluorescent live cell imaging it is critical to keep excitation light dose as low as possible, especially in the presence of photosensitizer drugs, which generate free radicals upon photobleaching. During fluorescent imaging, stress by excitation and free radicals induces serious cell damages that may arrest the cell cycle. This limits the usefulness of the technique for drug discovery, when prolonged live cell imaging is necessary. This paper presents a strategy to provide gentle experimental conditions for dynamic monitoring of the proliferation of human lung epithelial carcinoma cells (A549) in the presence of the photosensitizer Polyvinylpyrrolidone-Hypericin. The distinctive strategy of this paper is based on the stringent environmental control and optimizing the excitation light dose by (i) using a low-power pulsed blue light-emitting diode with short pulse duration of 1.29 ms and (ii) adding a nontoxic fluorescent dye called carboxyfluorescein-diacetate-succinimidyl-ester (CFSE) to improve the fluorescence signals. To demonstrate the usefulness of the strategy, fluorescence signals and proliferation of dual-marked cells, during 5-h fluorescence imaging under pulsed excitation, were compared with those kept under continuous excitation and nonmarked reference cells. The results demonstrated 3% cell division and 2% apoptosis due to pulsed excitation compared to no division and 85% apoptosis under the continuous irradiation. Therefore, our strategy allows live cell imaging to be performed over longer time scales than with conventional continuous excitation. PMID:21974829

Penjweini, Rozhin; Loew, Hans G.; Hamblin, Michael R.; Kratky, Karl W.

2011-01-01

261

New fluorescence reactions in DNA cytochemistry. 2. Microscopic and spectroscopic studies on fluorescent aluminum complexes  

SciTech Connect

Metal-dye complexes are widely applied in light microscopic techniques for chromatin staining (e.g., hematoxylin and carmine), but fluorescent complexes between phosphate-binding cations and suitable ligands have been little used. Preformed and postformed Al complexes with different anionic dyes induced strong and selective fluorescence reactions in nuclei from chicken blood smears, frozen sections, paraffin-embedded sections and Epon-embedded sections of mouse and rat tissues, mitotic chromosomes, meiotic chromosomes and kinetoplasts of Trypanosoma cruzi epimastigotes. The DNA-dependent fluorescence of these structures showed a very low fading rate. The emission colors were related to the ligand. The most suitable compounds for forming fluorescent Al chelates were 8-hydroxyquinoline, morin, nuclear fast red and purpurin. Staining with diluted carmine solutions and InCl3 mordanting, followed by 8-hydroxyquinoline, also induced chromatin fluorescence. After treating isolated mouse chromosomes with the preformed complex Al-nuclear fast red, x-ray microanalysis indicated a P:Al:dye binding ratio of about 40:15:1. The selectivity, stability and easy formation of these fluorescent Al complexes are obvious advantages for their use as new cytochemical probes in cytologic studies.

Del Castillo, P.; Llorente, A.R.; Gomez, A.; Gosalvez, J.; Goyanes, V.J.; Stockert, J.C. (Autonomous Univ., Madrid (Spain))

1990-02-01

262

Purified azure B as a reticulocyte stain.  

PubMed Central

A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two dyes, varying amounts of an extraneous, particulate dye deposit were present in these preparations, making accuracte counting both tedious and timeconsuming. Purified azure B, on the other hand, gave reproducibly stained, deposit-free preparations. Reticulocyte counts obtained from azure B preparations correlated almost exactly with those determined using new methylene blue. Purified azure B is therefore recommended as a convenient reticulocyte stain for routine use. Images PMID:64475

Marshall, P N; Bentley, S A; Lewis, S M

1976-01-01

263

Labeling the oily core of nanocapsules and lipid-core nanocapsules with a triglyceride conjugated to a fluorescent dye as a strategy to particle tracking in biological studies.  

PubMed

The synthesis of novel fluorescent materials represents a very important step to obtain labeled nanoformulations in order to evaluate their biological behavior. The strategy of conjugating a fluorescent dye with triacylglycerol allows that either particles differing regarding supramolecular structure, i.e., nanoemulsions, nanocapsules, lipid-core nanocapsules, or surface charge, i.e., cationic nanocapsules and anionic nanocapsules, can be tracked using the same labeled material. In this way, a rhodamine B-conjugated triglyceride was obtained to prepare fluorescent polymeric nanocapsules. Different formulations were obtained, nanocapsules (NC) or lipid-core nanocapsules (LNC), using the labeled oil and Eudragit RS100, Eudragit S100, or poly(caprolactone) (PCL), respectively. The rhodamine B was coupled with the ricinolein by activating the carboxylic function using a carbodiimide derivative. Thin layer chromatography, proton nuclear magnetic resonance ((1)H-NMR), Fourier transform infrared spectroscopy (FTIR), UV-vis, and fluorescence spectroscopy were used to identify the new product. Fluorescent nanocapsule aqueous suspensions were prepared by the solvent displacement method. Their pH values were 4.6 (NC-RS100), 3.5 (NC-S100), and 5.0 (LNC-PCL). The volume-weighted mean diameter (D 4.3) and polydispersity values were 150 nm and 1.05 (NC-RS100), 350 nm and 2.28 (NC-S100), and 270 nm and 1.67 (LNC-PCL). The mean diameters determined by photon correlation spectroscopy (PCS) (z-average) were around 200 nm. The zeta potential values were +5.85 mV (NC-RS100), -21.12 mV (NC-S100), and -19.25 mV (LNC-PCL). The wavelengths of maximum fluorescence emission were 567 nm (NC-RS100 and LNC-PCL) and 574 nm (NC-S100). Fluorescence microscopy was used to evaluate the cell uptake (human macrophage cell line) of the fluorescent nanocapsules in order to show the applicability of the approach. When the cells were treated with the fluorescent nanocapsules, red emission was detected around the cell nucleus. We demonstrated that the rhodamine B-conjugated triglyceride is a promising new material to obtain versatile dye-labeled nanocarriers presenting different chemical nature in their surfaces. PMID:24936156

Fiel, Luana Almeida; Contri, Renata Vidor; Bica, Juliane Freitas; Figueiró, Fabrício; Battastini, Ana Maria Oliveira; Guterres, Sílvia Stanisçuaski; Pohlmann, Adriana Raffin

2014-01-01

264

Identification of cervidae DNA in feedstuff using a real-time polymerase chain reaction method with the new fluorescence intercalating dye EvaGreen.  

PubMed

A real-time qualitative and quantitative polymerase chain reaction method (cer-194) using the fluorescence dye EvaGreen and aimed at the cytochrome b sequence was established for detection of cervidae DNA in feedstuff. Eight meat meal samples derived from deer, bovine, ovine, camel, pig, rabbit, fish, and chicken and 17 cervidae hair samples covering 2 subfamilies, 4 genera, and 7 species were tested to prove the specificity of the cer-194 system and its universality within the cervidae family. Detection limit of 0.1% deer meat in fish meal, blood powder, and feather powder matrixes was confirmed. PMID:19382576

Chen, Ying; Wu, Yajun; Wang, Jing; Xu, Baoliang; Zhong, Zhengyu; Xia, Jingshi

2009-01-01

265

DEVELOPMENT AND CALIBRATION OF A TWO-DYE FLUORESCENCE SYSTEM FOR USE IN TWO-PHASE MICRO FLOW THERMOMETRY  

E-print Network

. Applications of such cooling strategies include the use of heat pipes [2], micro-jet impingement [3], micro fraction Extinction coefficient of water [1/m] a Molar extinction coefficient of the dye [L/mol-m] c Molar

Hidrovo, Carlos H.

266

Bench-Top Antigen Detection Technique that Utilizes Nanofiltration and Fluorescent Dyes which Emit and Absorb Light in the Near Infrared  

NASA Technical Reports Server (NTRS)

This article discusses the development of a bench-top technique to detect antigens in fluids. The technique involves the use of near infrared NIR fluorescent dyes conjugated to antibodies, centrifugation, nanofilters, and spectrometry. The system used to detect the antigens utilizes a spectrometer, fiber optic cables, NIR laser, and laptop computer thus making it portable and ideally suited for desk top analysis. Using IgM as an antigen and the secondary antibody, anti-IgM conjugated to the near infrared dye, IRDye (trademark) 800, for detection, we show that nanofiltration can efficiently and specifically separate antibody-antigen complexes in solution and that the complexes can be detected by a spectrometer and software using NIR laser excitation at 778 nm and NIR dye offset emission at 804 nm. The peak power detected at 778 nm for the excitation emission and at 804 nm for the offset emission is 879 pW (-60.06 dBm) and 35.7 pW (-74.5 dBm), respectively.

Varaljay-Spence, Vanessa A.; Scardelletti, Maximilian C.

2007-01-01

267

Organic fluorescent dyes supported on activated boron nitride: a promising blue light excited phosphors for high-performance white light-emitting diodes.  

PubMed

We report an effective and rare-earth free light conversion material synthesized via a facile fabrication route, in which organic fluorescent dyes, i.e. Rhodamine B (RhB) and fluorescein isothiocyanate (FITC) are embedded into activated boron nitride (?BN) to form a composite phosphor. The composite phosphor shows highly efficient Förster resonance energy transfer and greatly improved thermal stability, and can emit at broad visible wavelengths of 500-650?nm under the 466?nm blue-light excitation. By packaging of the composite phosphors and a blue light-emitting diode (LED) chip with transparent epoxy resin, white LED with excellent thermal conductivity, current stability and optical performance can be realized, i.e. a thermal conductivity of 0.36?W/mk, a Commission Internationale de 1'Eclairage color coordinates of (0.32, 0.34), and a luminous efficiency of 21.6?lm·W(-1). Our research opens the door toward to the practical long-life organic fluorescent dyes-based white LEDs. PMID:25682730

Li, Jie; Lin, Jing; Huang, Yang; Xu, Xuewen; Liu, Zhenya; Xue, Yanming; Ding, Xiaoxia; Luo, Han; Jin, Peng; Zhang, Jun; Zou, Jin; Tang, Chengchun

2015-01-01

268

Vinyl-triphenylamine dyes, a new family of switchable fluorescent probes for targeted two-photon cellular imaging: from DNA to protein labeling.  

PubMed

On the basis of our previous work on vinyl-triphenylamine derived DNA fluorophores we explored the structure space around this core by coupling it to diverse cationic, anionic and zwitterionic groups in the aim of targeting different classes of biomolecules. In parallel core modifications were performed to optimize the photophysical properties (quantum yield, two-photon absorption). The resulting water soluble ?-conjugated molecules are called TP dyes and display an exceptional combination of optical properties: high two-photon absorption cross-section, high photostability, no self-quenching, and switchable fluorescence emission when bound to a biopolymer matrix. The linear and nonlinear optical properties of the TP dyes were studied in vitro in presence of DNA and in presence of a model protein (human serum albumin) using complementary absorption and fluorescence spectroscopy characterization tools. Structure modifications enabled to switch from DNA probes (cationic TP-pyridinium series) to protein probes (anionic TP-rhodanine series) without affecting the optical properties. Finally most TP compounds appear cell-permeant and show an intracellular localization consistent with their in vitro target specificity. PMID:22614341

Dumat, Blaise; Bordeau, Guillaume; Aranda, Ana I; Mahuteau-Betzer, Florence; El Harfouch, Yara; Metgé, Germain; Charra, Fabrice; Fiorini-Debuisschert, Céline; Teulade-Fichou, Marie-Paule

2012-08-14

269

In vivo tumor-targeted dual-modal fluorescence/CT imaging using a nanoprobe co-loaded with an aggregation-induced emission dye and gold nanoparticles.  

PubMed

As an intensely studied computed tomography (CT) contrast agent, gold nanoparticle has been suggested to be combined with fluorescence imaging modality to offset the low sensitivity of CT. However, the strong quenching of gold nanoparticle on fluorescent dyes requires complicated design and shielding to overcome. Herein, we report a unique nanoprobe (M-NPAPF-Au) co-loading an aggregation-induced emission (AIE) red dye and gold nanoparticles into DSPE-PEG2000 micelles for dual-modal fluorescence/CT imaging. The nanoprobe was prepared based on a facile method of "one-pot ultrasonic emulsification". Surprisingly, in the micelles system, fluorescence dye (NPAPF) efficiently overcame the strong fluorescence quenching of shielding-free gold nanoparticles and retained the crucial AIE feature. In vivo studies demonstrated the nanoprobe had superior tumor-targeting ability, excellent fluorescence and CT imaging effects. The totality of present studies clearly indicates the significant potential application of M-NPAPF-Au as a dual-modal non-invasive fluorescence/X-ray CT nanoprobe for in vivo tumor-targeted imaging and diagnosis. PMID:25542798

Zhang, Jimei; Li, Chan; Zhang, Xu; Huo, Shuaidong; Jin, Shubin; An, Fei-Fei; Wang, Xiaodan; Xue, Xiangdong; Okeke, C I; Duan, Guiyun; Guo, Fengguang; Zhang, Xiaohong; Hao, Jifu; Wang, Paul C; Zhang, Jinchao; Liang, Xing-Jie

2015-02-01

270

Comparison of the Sequence-Dependent Fluorescence of the Cyanine Dyes Cy3, Cy5, DyLight DY547 and DyLight DY647 on Single-Stranded DNA  

PubMed Central

Cyanine dyes are commonly used for fluorescent labeling of DNA and RNA oligonucleotides in applications including qPCR, sequencing, fluorescence in situ hybridization, Förster resonance energy transfer, and labeling for microarray hybridization. Previous research has shown that the fluorescence efficiency of Cy3 and Cy5, covalently attached to the 5? end of single-stranded DNA, is strongly sequence dependent. Here, we show that DY547 and DY647, two alternative cyanine dyes that are becoming widely used for nucleic acid labeling, have a similar pattern of sequence-dependence, with adjacent purines resulting in higher intensity and adjacent cytosines resulting in lower intensity. Investigated over the range of all 1024 possible DNA 5mers, the intensities of Cy3 and Cy5 drop by ?50% and ?65% with respect to their maxima, respectively, whereas the intensities of DY547 and DY647 fall by ?45% and ?40%, respectively. The reduced magnitude of change of the fluorescence intensity of the DyLight dyes, particularly of DY647 in comparison with Cy5, suggests that these dyes are less likely to introduce sequence-dependent bias into experiments based on fluorescent labeling of nucleic acids. PMID:24454899

Kretschy, Nicole; Somoza, Mark M.

2014-01-01

271

DUAL STAINING OF NATURAL BACTERIOPLANKTON WITH 4,6-DIAMINDO-2-PHENYLINDOLE AND FLUORESCENT OLIGONUCLEOTIDE PROBES TARGETING KINGDOM-LEVEL 16S RRNA SEQUENCES  

EPA Science Inventory

A method for quantifying eubacterial cell densities in dilute communities of small bacterioplankton is presented. ells in water samples were stained with 4',6-diamidino-2-phenylindole (DAPI), transferred to gelatin-coated slides, and hybridized with rhodamine-labeled oligonucleot...

272

Fluorescent styryl dyes FM1-43 and FM2-10 are muscarinic receptor antagonists: intravital visualization of receptor occupancy.  

PubMed

The fluorescent styryl dyes FM1-43 and FM2-10 have been used to visualize the endocytic and exocytic processes involved in neurotransmission in a variety of central and peripheral nerve preparations. Their utility is limited to some extent by a poorly understood vesicular-independent labelling of cells and tissues. We show here that one likely cause of this troublesome background labelling is that FM1-43 and FM2-10 are selective and competitive antagonists at both cloned and endogenously expressed muscarinic acetylcholine receptors. In radioligand binding studies, FM1-43 and FM2-10 bound with moderate affinity (23-220 nM) to membranes of Chinese hamster ovary (CHO) cells expressing cloned human muscarinic receptors (M1-M5). In functional studies in vitro, FM1-43 and FM2-10 inhibited electrical field stimulation (EFS) and acetylcholine-induced cholinergic contractions of guinea-pig tracheal strips (IC50: FM1-43, 0.4 +/- 0.1; FM2-10, 1.6 +/- 0.1 microM; concentration of antagonist producing a 2-fold leftward shift in the acetylcholine concentration-response curve (Kb): FM1-43, 0.3 +/- 0.1; FM2-10, 15.8 +/- 10.1 microM). Neither compound inhibited EFS-evoked, non-adrenergic non-cholinergic nerve-mediated relaxations or contractions of the airways, or contractions mediated by histamine H1 receptor or tachykinin NK2 receptor activation. Incubating freshly excised tracheal whole-mount preparations with 5 microM FM1-43 resulted in intense fluorescence labelling of the smooth muscle that was reduced by up to 90% in the presence of selective M2 and M3 receptor antagonists. The potency of the FM dyes as muscarinic receptor antagonists is within the concentration range used to study vesicular cycling at nerve terminals. Given that muscarinic receptors play a key role in the regulation of neurotransmitter release from a variety of neurones, the anticholinergic properties of FM dyes may have important implications when studying vesicular events in the nervous system. In addition, these dyes may provide a novel tool for visualizing muscarinic receptor occupancy in living tissue or cell preparations. PMID:16728454

Mazzone, Stuart B; Mori, Nanako; Burman, Miriam; Palovich, Michael; Belmonte, Kristen E; Canning, Brendan J

2006-08-15

273

A symmetrical fluorous dendron-cyanine dye-conjugated bimodal nanoprobe for quantitative 19F MRI and NIR fluorescence bioimaging.  

PubMed

(19)F MRI and optical imaging are two powerful noninvasive molecular imaging modalities in biomedical applications. (19)F MRI has great potential for high resolution in vivo imaging, while fluorescent probes enable ultracontrast cellular/tissue imaging with high accuracy and sensitivity. A bimodal nanoprobe is developed, integrating the merits of (19)F MRI and fluorescence imaging into a single synthetic molecule, which is further engineered into nanoprobe, by addressing shortcomings of conventional contrast agents to explore the quantitative (19)F MRI and fluorescence imaging and cell tracking. Results show that this bimodal imaging nanoprobe presents high correlation of (19)F MR signal and NIR fluorescence intensity in vitro and in vivo. Additionally, this nanoprobe enables quantitative (19)F MR analysis, confirmed by a complementary fluorescence analysis. This unique feature can hardly be obtained by traditional (19)F MRI contrast agents. It is envisioned that this nanoprobe can hold great potential for quantitative and sensitive multi-modal molecular imaging. PMID:24789108

Wang, Zhe; Yue, Xuyi; Wang, Yu; Qian, Chunqi; Huang, Peng; Lizak, Marty; Niu, Gang; Wang, Fu; Rong, Pengfei; Kiesewetter, Dale O; Ma, Ying; Chen, Xiaoyuan

2014-08-01

274

DNA fragment sizing and sorting by laser-induced fluorescence  

DOEpatents

A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.

Hammond, Mark L. (Angier, NC); Jett, James H. (Los Alamos, NM); Keller, Richard A. (Los Alamos, NM); Marrone, Babetta L. (Los Alamos, NM); Martin, John C. (Los Alamos, NM)

1996-01-01

275

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes.  

PubMed

Human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM)-based assays are emerging as a promising tool for the in vitro preclinical screening of QT interval-prolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. To test the precision and applicability of the near-infrared fluorescent voltage-sensitive dye 1-(4-sulfanatobutyl)-4-{?[2-(di-n-butylamino)-6-naphthyl]butadienyl}quinolinium betaine (di-4-ANBDQBS) for moderate-throughput electrophysiological analyses, we compared simultaneous transmembrane voltage and optical action potential (AP) recordings in human iPSC-CM loaded with di-4-ANBDQBS. Optical AP recordings tracked transmembrane voltage with high precision, generating nearly identical values for AP duration (AP durations at 10%, 50%, and 90% repolarization). Human iPSC-CMs tolerated repeated laser exposure, with stable optical AP parameters recorded over a 30-min study period. Optical AP recordings appropriately tracked changes in repolarization induced by pharmacological manipulation. Finally, di-4-ANBDQBS allowed for moderate-throughput analyses, increasing throughput >10-fold over the traditional patch-clamp technique. We conclude that the voltage-sensitive dye di-4-ANBDQBS allows for high-precision optical AP measurements that markedly increase the throughput for electrophysiological characterization of human iPSC-CMs. PMID:25172899

Lopez-Izquierdo, Angelica; Warren, Mark; Riedel, Michael; Cho, Scott; Lai, Shuping; Lux, Robert L; Spitzer, Kenneth W; Benjamin, Ivor J; Tristani-Firouzi, Martin; Jou, Chuanchau J

2014-11-01

276

The antioxidant glutathione in the fish cell lines EPC and BCF-2: response to model pro-oxidants as measured by three different fluorescent dyes.  

PubMed

Reduced glutathione (GSH) protects cells against injury by oxidative stress and maintains a range of vital functions. In vitro cell cultures have been used as experimental models to study the role of GSH in chemical toxicity in mammals; however, this approach has been rarely used with fish cells to date. The present study aimed to evaluate sensitivity and specificity of three fluorescent dyes for measuring pro-oxidant-induced changes of GSH contents in fish cell lines: monochlorobimane (mBCl), 5-chloromethylfluorescein diacetate (CMFDA) and 7-amino-4-chloromethylcoumarin (CMAC-blue). Two cell lines were studied, the EPC line established from a skin tumour of carp Cyprinus carpio, and BF-2 cells established from fins of bluegill sunfish Lepomis macrochirus. The cells were exposed for 6 and 24 h to low cytotoxic concentrations of pro-oxidants including hydrogen peroxide, paraquat (PQ), copper and the GSH synthesis inhibitor, L-buthionine-SR-sulfoximine (BSO). The results indicate moderate differences in the GSH response between EPC and BF-2 cells, but distinct differences in the magnitude of the GSH response for the four pro-oxidants. Further, the choice of GSH dye can critically affect the results, with CMFDA appearing to be less specific for GSH than mBCl and CMAC-blue. PMID:19444932

Jos, A; Cameán, A M; Pflugmacher, S; Segner, H

2009-04-01

277

A review of the chemistry and uses of crocins and crocetin, the carotenoid natural dyes in saffron, with particular emphasis on applications as colorants including their use as biological stains.  

PubMed

The perennial flowering plant, saffron crocus (Crocus sativus L.), is the source of the most expensive spice in the world. The dried stigmas of saffron flowers are the source of a natural dye, saffron, which has been used from ancient times for dyeing silk and fabric rugs, and for painting; it also has been used for cooking and in medicine. The yellow compounds present in the dye include crocins, which are 20-carbon water soluble glycosyl derivatives of the carotenoid, crocetin, and the dicarboxylic acid itself. We review the chemistry of these compounds and discuss various applications of saffron as a natural dye. We review in particular the use of saffron or its constituents in histopathologic techniques. PMID:24665936

Bathaie, S Z; Farajzade, A; Hoshyar, R

2014-08-01

278

Joint fluid Gram stain  

MedlinePLUS

Gram stain of joint fluid ... result means no bacteria are present on the Gram stain. Note: Normal value ranges may vary slightly ... Abnormal results mean bacteria were seen on the Gram stain. This may be a sign of a ...

279

pH-Induced Vesicle-to-Micelle Transition in Amphiphilic Diblock Copolymer: Investigation by Energy Transfer between in Situ Formed Polymer Embedded Gold Nanoparticles and Fluorescent Dye.  

PubMed

The ability to regulate the formation of nanostructures through self-assembly of amphiphilic block copolymers is of immense significance in the field of biology and medicine. In this work, a new block copolymer synthesized by using reversible addition-fragmentation chain transfer (RAFT) polymerization technique from poly(ethylene glycol) monomethyl ether acrylate (PEGMA) and Boc-l-tryptophan acryloyloxyethyl ester (Boc-l-trp-HEA) was found to spontaneously form pH-responsive water-soluble nanostructures after removal of the Boc group. While polymer vesicles or polymerosomes were formed at physiological pH, the micelles were formed at acidic pH (< 5.2), and this facilitated a pH-induced reversible vesicle-to-micelle transition. Formation of these nanostructures was confirmed by different characterization techniques, viz. transmission electron microscopy, dynamic light scattering, and steady-state fluorescence measurements. Further, these vesicles were successfully utilized to reduce HAuCl4 and stabilize the resulting gold nanoparticles (AuNPs). These AuNPs, confined within the hydrophobic shell of the vesicles, could participate in energy transfer process with fluorescent dye molecules encapsulated in the core of the vesicles, thus forming a nanometal surface energy transfer (NSET) pair. Subsequently, following the efficiency of energy transfer between this pair, it was possible to monitor the process of transition from vesicles to micelles. Thus, in this work, we have successfully demonstrated that NSET can be used to follow the transition between nanostructures formed by amphiphilic block copolymers. PMID:25494810

Maiti, Chiranjit; Banerjee, Rakesh; Maiti, Saikat; Dhara, Dibakar

2015-01-13

280

Analysis of Ground-Water Flow in the Madison Aquifer using Fluorescent Dyes Injected in Spring Creek and Rapid Creek near Rapid City, South Dakota, 2003-04  

USGS Publications Warehouse

The Madison aquifer, which contains fractures and solution openings in the Madison Limestone, is used extensively for water supplies for the city of Rapid City and other suburban communities in the Rapid City, S. Dak., area. The 48 square-mile study area includes the west-central and southwest parts of Rapid City and the outcrops of the Madison Limestone extending from south of Spring Creek to north of Rapid Creek. Recharge to the Madison Limestone occurs when streams lose flow as they cross the outcrop. The maximum net loss rate for Spring and Rapid Creek loss zones are 21 and 10 cubic feet per second (ft3/s), respectively. During 2003 and 2004, fluorescent dyes were injected in the Spring and Rapid Creek loss zones to estimate approximate locations of preferential flow paths in the Madison aquifer and to measure the response and transit times at wells and springs. Four injections of about 2 kilograms of fluorescein dye were made in the Spring Creek loss zone during 2003 (sites S1, S2, and S3) and 2004 (site S4). Injection at site S1 was made in streamflow just upstream from the loss zone over a 12-hour period when streamflow was about equal to the maximum loss rate. Injections at sites S2, S3, and S4 were made in specific swallow holes located in the Spring Creek loss zone. Injection at site R1 in 2004 of 3.5 kilograms of Rhodamine WT dye was made in streamflow just upstream from the Rapid Creek loss zone over about a 28-hour period. Selected combinations of 27 wells, 6 springs, and 3 stream sites were monitored with discrete samples following the injections. For injections at sites S1-S3, when Spring Creek streamflow was greater than or equal to 20 ft3/s, fluorescein was detected in samples from five wells that were located as much as about 2 miles from the loss zone. Time to first arrival (injection at site S1) ranged from less than 1 to less than 10 days. The maximum fluorescein concentration (injection at site S1) of 120 micrograms per liter (ug/L) at well CO, which is located adjacent to the loss zone, was similar to the concentration in the stream. Fluorescein arrived at well NON (injection at site S1), which is located about 2 miles northeast of the loss zone, within about 1.6 days, and the maximum concentration was 44 ug/L. For injection at site S4, when streamflow was about 12 ft3/s, fluorescein was detected in samples from six wells and time to first arrival ranged from 0.2 to 16 days. Following injection at site S4 in 2004, the length of time that dye remained in the capture zone of well NON, which is located approximately 2 miles from the loss zone, was almost an order of magnitude greater than in 2003. For injection at site R1, Rhodamine WT was detected at well DRU and spring TI-SP with time to first arrival of about 0.5 and 1.1 days and maximum concentrations of 6.2 and 0.91 ug/L, respectively. Well DRU and spring TI-SP are located near the center of the Rapid Creek loss zone where the creek has a large meander. Measurable concentrations were observed for spring TI-SP as many as 109 days after the dye injection. The direction of a conduit flow path in the Spring Creek area was to the northeast with ground-water velocities that ranged from 770 to 6,500 feet per day. In the Rapid Creek loss zone, a conduit flow path east of the loss zone was not evident from the dye injection.

Putnam, Larry D.; Long, Andrew J.

2007-01-01

281

Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label  

Technology Transfer Automated Retrieval System (TEKTRAN)

Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as antibody labels to increase the fluorescence signal and sensitivity. Ep...

282

Ratiometric fluorescence detection of cysteine and homocysteine with a BODIPY dye by mimicking the native chemical ligation.  

PubMed

The selective detection of cysteine and homocysteine over glutathione and other amino acids was demonstrated with an 8-MeS-BODIPY probe by mimicking the native chemical ligation approach, which allowed the selective and ratiometric fluorescence sensing of cysteine over other biothiols at physiologically relevant concentrations and in different organs of a zebrafish. PMID:25426498

Ma, Dong Hee; Kim, Dokyoung; Seo, Eunseok; Lee, Sang-Joon; Ahn, Kyo Han

2014-12-15

283

Selective recognition of parallel and anti-parallel thrombin-binding aptamer G-quadruplexes by different fluorescent dyes.  

PubMed

G-quadruplexes (G4) have been found increasing potential in applications, such as molecular therapeutics, diagnostics and sensing. Both Thio?avin T (ThT) and N-Methyl mesoporphyrin IX (NMM) become fluorescent in the presence of most G4, but thrombin-binding aptamer (TBA) has been reported as the only exception of the known G4-forming oligonucleotides when ThT is used as a high-throughput assay to identify G4 formation. Here, we investigate the interactions between ThT/NMM and TBA through fluorescence spectroscopy, circular dichroism and molecular docking simulation experiments in the absence or presence of cations. The results display that a large ThT ?uorescence enhancement can be observed only when ThT bind to the parallel TBA quadruplex, which is induced to form by ThT in the absence of cations. On the other hand, great promotion in NMM fluorescence can be obtained only in the presence of anti-parallel TBA quadruplex, which is induced to fold by K+ or thrombin. The highly selective recognition of TBA quadruplex with different topologies by the two probes may be useful to investigate the interactions between conformation-specific G4 and the associated proteins, and could also be applied in label-free fluorescent sensing of other biomolecules. PMID:25245945

Zhao, Dan; Dong, Xiongwei; Jiang, Nan; Zhang, Dan; Liu, Changlin

2015-01-01

284

LysoTracker staining to aid in monitoring autophagy in Drosophila.  

PubMed

LysoTracker is an acidotropic dye that stains cellular acidic compartments, including lysosomes and autolysosomes. LysoTracker has been used to detect autophagy-associated lysosomal activity in Drosophila tissues including the fat body, midgut, salivary gland and ovary, as well as in Drosophila cell culture. A low level of LysoTracker staining can be observed under resting or well-fed conditions, and is increased following autophagic stimuli such as starvation. Here we provide a protocol for examining LysoTracker levels in Drosophila cultured cells in vitro using standard cell culture methods and flow cytometry. We also describe how to examine LysoTracker in fixed and nonfixed Drosophila tissues using fluorescence microscopy. Ovary tissue is used as an example. Dissections of ovaries are relatively easy to perform, given their large size. PMID:25183815

DeVorkin, Lindsay; Gorski, Sharon M

2014-09-01

285

Pontentiometric Cyanine Dyes Are Sensitive Probes for Mitochondria in Intact Plant Cells 1  

PubMed Central

Selected fluorescent dyes were tested for uptake by mitochrondria in intact cells of barley, maize, and onion. The cationic cyanine dye 3,3?-diheptyloxacarbocyanine iodide [DiOC7(3)] accumulated in mitochondria within 15 to 30 minutes without appreciable staining of other protoplasmic constituents. The number, shape, and movement of the fluorescent mitochondria could be seen readily, and the fluorescence intensity of the mitochondria could be monitored with a microscope photometer. Fluorescence was eliminated in 1 to 5 minutes by the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) indicating that maintenance of dye concentration was dependent on the inside-negative transmembrane potential maintained by functional mitochondria. Fluorescence of prestained mitochondria was enhanced within 5 to 10 minutes after addition of 0.1 millimolar kinetin to cells. The fluorescence in kinetintreated cells was dissipated by CCCP. These results suggest that kinetin interacted with respiratory processes resulting in higher potential across the mitochondrial membrane. Images Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:16665615

Liu, Zhanjiang; Bushnell, W. R.; Brambl, Robert

1987-01-01

286

Immunofluorescence staining of paraffin sections: creating DAB staining like virtual digital images using CMYK color conversion.  

PubMed

Crystal violet treatment of formalin fixed paraffin embedded tissue slides greatly reduces the endogenous autofluorescence, and allows immunofluorescence (IF) staining with FITC or Alexa488 conjugated antibodies. Using cold CCD camera to capture the fluorescence images makes this staining method very sensitive. Here we show that combination of IF with the simultaneous recording of crystal violet induced red and Hoechst 33258 induced blue fluorescence permits the localization of the IF signal over a cytoplasmic: nuclear red:blue stain that visualizes the microscopic anatomy of the underlying tissue. To make the visual interpretation of the IF staining easier for microscopists, who are used to DAB staining over weak hematoxilin-eosin background, we created a simple color conversion procedure that turns the captured three-color fluorescence RGB (red, green, blue) images over a black background into four color CMYK (cyan, magenta, yellow, key color (black)) images. PMID:19112433

Buchynska, L; Kashuba, E; Szekely, L

2008-12-01

287

7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2014 CFR

...2014-01-01 2014-01-01 false Wood and concrete stains. 3201.87 Section...PROCUREMENT Designated Items § 3201.87 Wood and concrete stains. (a) Definition...be applied as a finish for concrete and wood surfaces and that contain dyes or...

2014-01-01

288

7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2013 CFR

...2013-01-01 2013-01-01 false Wood and concrete stains. 3201.87 Section...PROCUREMENT Designated Items § 3201.87 Wood and concrete stains. (a) Definition...be applied as a finish for concrete and wood surfaces and that contain dyes or...

2013-01-01

289

Dye-coated europium monosulfide  

SciTech Connect

Nanoparticles of EuS were synthesized using europium dithiocarbamate complexes. The resulting nanoparticles were coated with the dye, 1-pyrene carboxylic acid and the resulting material was characterized using X-ray powder diffraction, TEM, and UV-visible spectroscopy. Fluorescence spectroscopy was used to determine the relative energy of the conduction band edge to the excited state energy of the dye. -- Graphical abstract: Dye sensitized magnetic semiconductor materials were prepared by synthesizing EuS nanoparticles using single source precursors and coating with the dye, 1-pyrene carboxylic acid. Display Omitted highlights: > Synthesized EuS nanoparticles, 11{+-}2.4 nm characterized using XRD, TEM, and UV-vis. spect. > Grafted a dye to the surface and characterized the product using XRD, FTIR, UV-vis., and TEM. > Studied the photophysical properties using fluorescence spectroscopy. > Determined the relative dye excited state to the conduction band of the semiconductor.

Kar, Srotoswini [Department of Chemistry, Georgetown University, Washington D.C. 20057 (United States); Dollahon, Norman R. [Department of Biology, Villanova University, Villanova, PA 19085 (United States); Stoll, Sarah L., E-mail: sls55@georgetown.ed [Department of Chemistry, Georgetown University, Washington D.C. 20057 (United States)

2011-05-15

290

Detection of chloroform in water using an azo dye-modified ?-cyclodextrin - Epichlorohydrin copolymer as a fluorescent probe  

NASA Astrophysics Data System (ADS)

Chlorination disinfection by-products (DBPs) in water pose a health threat to humans and the aquatic environment. Their detection in water sources is therefore vital. Herein we present the detection of chloroform, a DBP, using a molecular fluorescent probe. The detection was based on the quenching of fluorescence of the probe by chloroform due to host-guest complex formation between ?-cyclodextrin in the probe and the chloroform molecule. The stability constant for the host-guest complex was high at 3.825 × 104 M-1. Chloroform quenched the fluorescence of the copolymer the most compared to the other small chlorinated compounds studied, suggesting that the probe was more sensitive to chloroform, with a sensing factor of 0.35 compared to as low as 0.0073 for dichloromethane. There was no interference from other chloroalkanes on the quenching efficiency of chloroform. The probe was used to detect chloroform in dam water as well as in bottled water. Detection of chloroform in both water samples using the probe was possible without chemically treating the water samples which may introduce other pollutants.

Ncube, Phendukani; Krause, Rui W. M.; Mamba, Bhekie B.

291

Coffee stain on textiles. Mechanisms of staining and stain removal  

Microsoft Academic Search

Coffee stains on textiles are mainly caused by the water-soluble and acidic colored substances in coffee. The acidic nature\\u000a of coffee stain has been shown by ultraviolet and visible spectroscopy of coffee as a function of pH; ion-pair formation with\\u000a a cationic surfactant and titration with Hyamine 1622 and a surfactant-specific electrode; and precipitation of the colored\\u000a components in coffee

Erik Kissa

1995-01-01

292

The development of simple and sensitive small-molecule fluorescent probes for the detection of serum proteins after native polyacrylamide gel electrophoresis.  

PubMed

In this paper, a simple and sensitive small-molecule fluorescent probe, 2,5-dihydroxy-4'-dimethylaminochalcone (DHDMAC), was designed and synthesized for the detection of human serum proteins via hydrophobic interactions after polyacrylamide gel electrophoresis (PAGE). This probe produced lower fluorescence emission in the absence of proteins, and the emission intensity was significantly increased after the interaction with serum proteins. To demonstrate the imaging performance of this probe as a fluorescent dye, a series of experiments was conducted that included sensitivity comparison and 2D-PAGE. The results indicated that the sensitivity of DHDMAC staining is comparable to that of the most widely used fluorescent dye, SYPRO Ruby, and more protein spots (including thyroxine-binding globulin, angiotensinogen, afamin, zinc-?-2-glycoprotein and ?-1-antichymotrypsin) were detected after 2D-PAGE. Therefore, DHDMAC is a good protein reporter due to its fast staining procedure, low detection limits and high resolution. PMID:22475746

Wang, Fangfang; Huang, Lingyun; Na, Na; He, Dacheng; Sun, Dezhi; Ouyang, Jin

2012-05-21

293

Introduction of impermeable actin-staining molecules to mammalian cells by optoporation  

PubMed Central

The selective insertion of foreign materials, such as fluorescent markers or plasmids, into living cells has been a challenging problem in cell biology due to the cell membrane's selective permeability. However, it is often necessary that researchers insert such materials into cells for various dynamical and/or drug delivery studies. This problem becomes even more challenging if the study is to be limited to specific cells within a larger population, since other transfection methods, such as viral transfection and lipofection, are not realizable with a high degree of spatial selectivity. Here, we have used a focused femtosecond laser beam to create a small transient hole in the cellular membrane (optoporation) in order to inject nanomolar concentrations of rhodamine phalloidin (an impermeable dye molecule for staining filamentous actin) into targeted living mammalian cells (both HEK and primary cortical neurons). Following optoporation, the dye bound to the intracellular actin network and rise in fluorescence intensity was observed. Theoretical dynamics of the dye's diffusion is discussed, and numerical simulations of diffusion time constants are found to match well with experimental values. PMID:25315642

Dhakal, Kamal; Black, Bryan; Mohanty, Samarendra

2014-01-01

294

Introduction of impermeable actin-staining molecules to mammalian cells by optoporation.  

PubMed

The selective insertion of foreign materials, such as fluorescent markers or plasmids, into living cells has been a challenging problem in cell biology due to the cell membrane's selective permeability. However, it is often necessary that researchers insert such materials into cells for various dynamical and/or drug delivery studies. This problem becomes even more challenging if the study is to be limited to specific cells within a larger population, since other transfection methods, such as viral transfection and lipofection, are not realizable with a high degree of spatial selectivity. Here, we have used a focused femtosecond laser beam to create a small transient hole in the cellular membrane (optoporation) in order to inject nanomolar concentrations of rhodamine phalloidin (an impermeable dye molecule for staining filamentous actin) into targeted living mammalian cells (both HEK and primary cortical neurons). Following optoporation, the dye bound to the intracellular actin network and rise in fluorescence intensity was observed. Theoretical dynamics of the dye's diffusion is discussed, and numerical simulations of diffusion time constants are found to match well with experimental values. PMID:25315642

Dhakal, Kamal; Black, Bryan; Mohanty, Samarendra

2014-01-01

295

Clinical staining of the ocular surface: mechanisms and interpretations.  

PubMed

In this article we review the mechanism of ocular surface staining. Water-soluble dyes are excluded from the normal epithelium by tight junctions, the plasma membranes and the surface glycocalyx. Shed cells can take up dye. A proportion of normal corneas show sparse, scattered time-dependent, punctate fluorescein uptake, which, we hypothesise, is due to a graded loss of the glycocalyx barrier, permitting transcellular entry into pre-shed cells. In pathological staining, there is little evidence of 'micropooling' at sites of shedding and the term 'punctate erosion' may be a misnomer. It is more likely that the initial event involves transcellular dye entry and, in addition, diffusion across defective tight junctions. Different dye-staining characteristics probably reflect differences in molecular size and other physical properties of each dye, coupled with differences in visibility under the conditions of illumination used. This is most relevant to the rapid epithelial spread of fluorescein from sites of punctate staining, compared to the apparent confinement of dyes to staining cells with dyes such as lissamine green and rose bengal. We assume that fluorescein, with its lower molecular weight, spreads initially by a paracellular route and then by transcellular diffusion. Solution-Induced Corneal Staining (SICS), related to the use of certain contact lens care solutions, may have a different basis, involving the non-pathological uptake of cationic preservatives, such as biguanides, into epithelial membranes and secondary binding of the fluorescein anion. It is transient and may not imply corneal toxicity. Understanding the mechanism of staining is relevant to the standardisation of grading, to monitoring disease and to the conduct of clinical trials. PMID:25461622

Bron, A J; Argüeso, P; Irkec, M; Bright, F V

2015-01-01

296

Dye filled security seal  

DOEpatents

A security seal for providing an indication of unauthorized access to a sealed object includes an elongate member to be entwined in the object such that access is denied unless the member is removed. The elongate member has a hollow, pressurizable chamber extending throughout its length that is filled with a permanent dye under greater than atmospheric pressure. Attempts to cut the member and weld it together are revealed when dye flows through a rupture in the chamber wall and stains the outside surface of the member.

Wilson, Dennis C. W. (Tijeras, NM)

1982-04-27

297

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye  

PubMed Central

An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to < 10 copies of target sequence in a plasmid and up to a 10-2 dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation. PMID:24323513

Watts, Matthew R.; James, Gregory; Sultana, Yasmin; Ginn, Andrew N.; Outhred, Alexander C.; Kong, Fanrong; Verweij, Jaco J.; Iredell, Jonathan R.; Chen, Sharon C-A.; Lee, Rogan

2014-01-01

298

A loop-mediated isothermal amplification (LAMP) assay for Strongyloides stercoralis in stool that uses a visual detection method with SYTO-82 fluorescent dye.  

PubMed

An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to < 10 copies of target sequence in a plasmid and up to a 10(-2) dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation. PMID:24323513

Watts, Matthew R; James, Gregory; Sultana, Yasmin; Ginn, Andrew N; Outhred, Alexander C; Kong, Fanrong; Verweij, Jaco J; Iredell, Jonathan R; Chen, Sharon C-A; Lee, Rogan

2014-02-01

299

[Physical-chemical properties of the mutant (protein) form of D-glucose/D-galactose-binding protein GGBP/H152C with an attached fluorescent dye BADAN].  

PubMed

The influence of various factors on the physico-chemical characteristics and complexation of glucose with a mutant form of D-glucose/D-galactose-binding protein which can be regarded as a sensor of the glucometer, namely the protein GGBP/H152C with solvatochromic dye BADAN attached to the cysteine residue Cys 152, has been investigated. The point mutation His 152Cys and attaching BADAN reduced the affinity of the mutant form GGBP/H152C to glucose more than 8-fold compared to the wild type protein. This allows using this mutant for the determination of sugar content in biological fluids extracted by transdermal technologies. Sufficiently rapid complexation of GGBP/H152C with glucose (the time of protein-glucose complex formation is not more than three seconds even in solutions with a viscosity of 4 cP) provides timely monitoring changes in the concentration of sugar. The changes of ionic strength and pH within the physiological range of values of these variables do not have significant influence on fluorescent characteristics of GGBP/H152C-BADAN. At acidic pH, (see symbol) some of the molecules GGBP/H152C is in the unfolded state. It has been shown that mutant form GGBP/H152C has relatively low resistance to guanidine hydrochloride denaturing effects. This result indicates the need for more stable proteins to create a sensor for glucose biosensor system. PMID:25474908

Fonin, A V; Stepanenko, O V; Povarova, O I; Volova, E A; Filippova, E M; Bublikov, G S; Kuznetsova, I M; Demchenko, A P; Turoverov, K K

2013-01-01

300

[Physical-chemical properties of the mutant (protein) form of D-glucose/D-galactose-binding protein GGBP/H152C with an attached fluorescent dye BADAN].  

PubMed

The influence of various factors on the physico-chemical characteristics and complexation of glucose with a mutant form of D-glucose/D-galactose-binding protein which can be regarded as a sensor of the glucometer, namely the protein GGBP/H152C with solvatochromic dye BADAN attached to the cysteine residue Cys 152, has been investigated. The point mutation His 152Cys and attaching BADAN reduced the affinity of the mutant form GGBP/H152C to glucose more than 8-fold compared to the wild type protein. This allows using this mutant for the determination of sugar content in biological fluids extracted by transdermal technologies. Sufficiently rapid complexation of GGBP/H152C with glucose (the time of protein-glucose complex formation is not more than three seconds even in solutions with a viscosity of 4 cP) provides timely monitoring changes in the concentration of sugar. The changes of ionic strength and pH within the physiological range of values of these variables do not have significant influence on fluorescent characteristics of GGBP/H152C-BADAN. At acidic pH, (see symbol) some of the molecules GGBP/H152C is in the unfolded state. It has been shown that mutant form GGBP/H152C has relatively low resistance to guanidine hydrochloride denaturing effects. This result indicates the need for more stable proteins to create a sensor for glucose biosensor system. PMID:25508687

2013-01-01

301

Spectral characteristics of the mutant form GGBP/H152C of D-glucose/D-galactose-binding protein labeled with fluorescent dye BADAN: influence of external factors  

PubMed Central

The mutant form GGBP/H152C of the D-glucose/D-galactose-binding protein with the solvatochromic dye BADAN linked to cysteine residue Cys 152 can be used as a potential base for a sensitive element of glucose biosensor system. We investigated the influence of various external factors on the physical-chemical properties of GGBP/H152C-BADAN and its complex with glucose. The high affinity (Kd = 8.5 µM) and high binding rate of glucose make GGBP/H152C-BADAN a good candidate to determine the sugar content in biological fluids extracted using transdermal techniques. It was shown that changes in the ionic strength and pH of solution within the physiological range did not have a significant influence on the fluorescent characteristics of GGBP/H152C-BADAN. The mutant form GGBP/H152C has relatively low resistance to denaturation action of GdnHCl and urea. This result emphasizes the need to find more stable proteins for the creation of a sensitive element for a glucose biosensor system. PMID:24711960

Fonin, Alexander V.; Stepanenko, Olga V.; Povarova, Olga I.; Volova, Catherine A.; Philippova, Elizaveta M.; Bublikov, Grigory S.; Kuznetsova, Irina M.; Demchenko, Alexander P.

2014-01-01

302

Port-Wine Stain  

MedlinePLUS

... related to port-wine stains are sometimes called salmon patches, which may also be called angel kisses ( ... of the baby's neck). Like port-wine stains, salmon patches start as flat, pink or red patches; ...

303

Stool Gram stain  

MedlinePLUS

... series of special stains are added to the sample. The lab team member looks at the stained smear under the microscope to check for bacteria. The color, size, and shape of the cells help identify the ...

304

Gram stain of urethral discharge  

MedlinePLUS

Urethral discharge Gram stain ... microscope slide. A series of stains called a gram stain is applied to the specimen. The stained ... culture ) should be performed in addition to the gram stain. More sophisticated diagnostic tests (such as PCR ...

305

Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections  

Microsoft Academic Search

Synopsis  Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency.Examination of tissue sections from 15

L. C. U. Junqueira; G. Bignolas; R. R. Brentani

1979-01-01

306

DNA fragment sizing and sorting by laser-induced fluorescence  

SciTech Connect

A method is provided for obtaining DNA fingerprints using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a selected sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is directly proportional to the fragment length. Additional dyes can be bound to the DNA piece and DNA fragments to provide information additional to length information. Oligonucleotide specific dyes and/or hybridization probes can be bound to the DNA fragments to provide information on oligonucleotide distribution or probe hybridization to DNA fragments of different sizes.

Jett, J.H.; Hammond, M.L.; Keller, R.A.; Marrone, B.L.; Martin, J.C.

1992-12-31

307

Detection Of Concrete Deterioration By Staining  

DOEpatents

A method using concentrated aqueous solutions of sodium cobaltinitrite and a rhodamine dye is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR), and to identify degraded concrete which results in a porous or semi-permeable paste due to carbonation or leaching. These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na--K--Ca--Si gels are identified by yellow staining, and alkali-poor, Ca--Si gels are identified by pink staining.

Guthrie, Jr., George D. (Santa Fe, NM); Carey, J. William (Santa Fe, NM)

1999-09-21

308

Laser treatment of port-wine stains  

PubMed Central

Port-wine stains are a type of capillary malformation affecting 0.3% to 0.5% of the population. Port-wine stains present at birth as pink to erythematous patches on the skin and/or mucosa. Without treatment, the patches typically darken with age and may eventually develop nodular thickening or associated pyogenic granuloma. Laser and light treatments provide improvement through selective destruction of vasculature. A variety of vascular-selective lasers may be employed, with the pulsed dye laser being the most common and well studied. Early treatment produces more optimal results. Advances in imaging and laser treatment technologies demonstrate potential to further improve clinical outcomes. PMID:25624768

Brightman, Lori A; Geronemus, Roy G; Reddy, Kavitha K

2015-01-01

309

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis  

PubMed Central

High Mechanical Index (MI) echocardiography with contrast agent has been shown to induce Evans blue staining of cardiomyocytes, seen one day after exposure, in addition to contraction band necrosis, seen immediately after exposure. This research examined the roles of necrosis vs. apoptosis in these bioeffects. Myocardial contrast echocardiography at high MI with 1:4 ECG triggering was performed in anesthetized rats at 1.5 MHz. Histologically observable cell injury was accumulated by infusing a high dose of 50 ?l/kg Definity® via tail vein for 5 min at the start of 10 min of scanning. Evans blue dye or propidium iodide was injected as an indicator of cardiomyocyte plasma membrane integrity. Histological sections were stained using the TUNEL method for labeling nuclei with DNA degradation (e.g. apoptosis). Evans blue fluorescent cells were counted on frozen sections or on hematoxylin-stained and TUNEL labeled paraffin sections. In addition, transmission electron microscopy was used to assess potential apoptotic nuclei. Hypercontraction and propidium iodide staining were observed immediately after imaging-exposure. Although TUNEL positive cells were evident after 4 h, these also had indications of contraction band necrosis and features of apoptosis were not confirmed by electron microscopy. Inflammatory cell infiltration was evident after 24 h. A second, more subtle injury was recognized by Evans blue staining, with minimal inflammatory cell infiltration at the morphologically intact stained cells after 24 h. Apoptosis was not detected by the TUNEL method in the cardiomyocytes stained with Evans blue at 24 h. However, Evans blue stained cell numbers declined after 48 h, with continued inflammatory cell infiltration. The initial insult for Evans blue stained cardiomyocytes apparently induced partial permeability of the plasma membrane, which led to gradual degeneration (but not apoptosis) and necrosis after 24–48 h. PMID:17689176

Miller, Douglas L.; Li, Peng; Dou, Chunyan; Armstrong, William F.; Gordon, David

2008-01-01

310

Voltage-Sensitive Dyes And Imaging Techniques Reveal New Patterns Of Electrical Activity In Heart Cortex  

NASA Astrophysics Data System (ADS)

Voltage-sensitive dyes bind to the plasms membrane of excitable cells (ie., muscle or nerve cells) and exhibit fluorescence and/or absorption changes that vary linearly with changes in transmembrane electrical potential. These potentiometric optical probes can be used to measure local changes in transmembrane potential by monitoring optical signals from dye molecules bound to the surface membrane. Consequently, when excitable cells are stained with such a dye and are stimulated to fire an electrical impulse (ie., an action potential (AP)), the changes in dye fluorescence have the characteristic shape and time course of APs recorded with an intracellular micro-electrode. Potentiometric dyes in conjuction with imaging techniques can now be used to visualize complex patterns and propagation of electrical activity. With photodiode arrays on video imaging techniques, patterns of biological electrical activity can be obtained with high temporal and spatial resolution which could not be obtained by conventional micro-electrodes. These methods reveal new details and offer powerful approaches to study fundamental problem in cardiac electrophysiology, communication in nerve networks, and the organization of cortical neurons.

Salama, Guy

1988-04-01

311

Bright, fluorescent dyes based on imidazo[1,2-a]pyridines that are capable of two-photon absorption.  

PubMed

A library of imidazo[1,2-a]pyridines was synthesized by using the Gevorgyan method and their linear and non-linear optical properties were studied. Derivatives that contained both electron-donating and electron-withdrawing groups at the 2?position were comprehensively investigated. Their emission quantum yield ranged between 0.2-0.7 and it was shown to depend on the substitution pattern, most notably that on the phenyl ring. Electron-donating substituents improved the luminescence performance of these compounds, whereas electron-withdrawing substituents led to a more erratic behavior. Substitution on the six-membered ring had less effect on the fluorescence properties. Extension of the delocalization increased the luminescence quantum yield. A new quadrupolar system was designed that contained two imidazo[1,2-a]pyridine units on its periphery and a 1,4-dicyanobenzene unit at its center. This system exhibited a large Stokes-shifted luminescence that was affected by the polarity and rigidity of the solvent, which was ascribed to emission from an excited state with strong charge-transfer character. This quadrupolar feature also led to an acceptable two-photon absorption response in the NIR region. PMID:23564658

Firmansyah, Dikhi; Ciuciu, Adina I; Hugues, Vincent; Blanchard-Desce, Mireille; Flamigni, Lucia; Gryko, Daniel T

2013-06-01

312

The Chemically Synthesized Ageladine A-Derivative LysoGlow84 Stains Lysosomes in Viable Mammalian Brain Cells and Specific Structures in the Marine Flatworm Macrostomum lignano.  

PubMed

Based on the chemical structure and the known chemical synthesis of the marine sponge alkaloid ageladine A, we synthesized the ageladine A-derivative 4-(naphthalene-2-yl)-1H-imidazo[4,5-c]pyridine trifluoroacetate (LysoGlow84). The two-step synthesis started with the Pictet-Spengler reaction of histamine and naphthalene-2-carbaldehyde to a tetrahydropyridine intermediate, which was dehydrogenated with activated manganese (IV) oxide to LysoGlow84. Structure and purity of the synthesized LysoGlow84 were confirmed by NMR spectroscopy and mass spectrometry. The fluorescence intensity emitted by LysoGlow84 depended strongly on the pH of the solvent with highest fluorescence intensity recorded at pH 4. The fluorescence maximum (at 315 nm excitation) was observed at 440 nm. Biocompatibility of LysoGlow84 was investigated using cultured rat brain astrocytes and the marine flatworm Macrostomum lignano. Exposure of the astrocytes for up to 6 h to micromolar concentrations of LysoGlow84 did not compromise cell viability, as demonstrated by several viability assays, but revealed a promising property of this compound for staining of cellular vesicles. Conventional fluorescence microscopy as well as confocal scanning microscopy of LysoGlow84-treated astrocytes revealed co-localization of LysoGlow84 fluorescence with that of LysoTracker® Red DND-99. LysoGlow84 stained unclear structures in Macrostomum lignano, which were identified as lysosomes by co-staining with LysoTracker. Strong fluorescence staining by LysoGlow84 was further observed around the worms' anterior gut and the female genital pore which were not counterstained by LysoTracker Red. Thus, LysoGlow84 is a new promising dye that stains lysosomes and other acidic compartments in cultured cells and in worms. PMID:25679913

Mordhorst, Thorsten; Awal, Sushil; Jordan, Sebastian; Petters, Charlotte; Sartoris, Linda; Dringen, Ralf; Bickmeyer, Ulf

2015-01-01

313

Polarized fluorescent nanospheres.  

PubMed

Fluorescent beads (nanoparticles, nanospheres) are commonly used in fluorescence spectroscopy and microscopy. Due to the random distribution of dye and high dye to nanoparticle ratio, the fluorescence polarization observed from the beads is low. Therefore beads are not used for polarization study. We demonstrate that photoselective bleaching creates beads with highly polarized fluorescence. First, the beads were immobilized in a PVA polymer. Second, the beads-doped PVA film was exposed to the illumination within the dye absorption band. A progressive decrease of absorption was observed. Next, photophysical properties of photobleached and not bleached films dissolved in water were compared. PMID:20389440

Luchowski, Rafal; Gryczynski, Zygmunt; Földes-Papp, Zeno; Chang, Aaron; Borejdo, Julian; Sarkar, Pabak; Gryczynski, Ignacy

2010-03-01

314

Twisted Cyanines: A Non-Planar Fluorogenic Dye with Superior Photostability and its Use in a Protein-Based Fluoromodule  

PubMed Central

The cyanine dye thiazole orange (TO) is a well-known fluorogenic stain for DNA and RNA, but this property precludes its use as an intracellular fluorescent probe for non-nucleic acid biomolecules. Further, as is the case with many cyanines, the dye suffers from low photostability. Here we report the synthesis of a bridge-substituted version of TO named ?-CN-TO, where the central methine hydrogen of TO is replaced by an electron withdrawing cyano group, which was expected to decrease the susceptibility of the dye toward singlet oxygen-mediated degradation. An X-ray crystal structure shows that ?-CN-TO is twisted drastically out of plane, in contrast to TO, which crystallizes in the planar conformation. ?-CN-TO retains the fluorogenic behavior of the parent dye TO in viscous glycerol/water solvent, but direct irradiation and indirect bleaching studies showed that ?-CN-TO is essentially inert to visible light and singlet oxygen. In addition, the twisted conformation of ?-CN-TO mitigates non-specific binding and fluorescence activation by DNA and a previously selected TO-binding protein and exhibits low background fluorescence in HeLa cell culture. ?-CN-TO was then used to select a new protein that binds and activates fluorescence from the dye. The new ?-CN-TO/protein fluoromodule exhibits superior photostability to an analogous TO/protein fluoromodule. These properties indicate that ?-CN-TO will be a useful fluorogenic dye in combination with specific RNA and protein binding partners for both in vitro and cell-based applications. More broadly, structural features that promote nonplanar conformations can provide an effective method for reducing nonspecific binding of cationic dyes to nucleic acids and other biomolecules. PMID:23252842

Shank, Nathaniel I.; Pham, Ha; Waggoner, Alan S.; Armitage, Bruce A.

2013-01-01

315

Dye Application, Manufacture of Dye Intermediates and Dyes  

NASA Astrophysics Data System (ADS)

It is difficult if not impossible to determine when mankind first systematically applied color to a textile substrate. The first colored fabrics were probably nonwoven felts painted in imitation of animal skins. The first dyeings were probably actually little more than stains from the juice of berries. Ancient Greek writers described painted fabrics worn by the tribes of Asia Minor. But just where did the ancient craft have its origins? Was there one original birthplace or were there a number of simultaneous beginnings around the world?

Freeman, H. S.; Mock, G. N.

316

Shedding light on the photostability of two intermolecular charge-transfer complexes between highly fluorescent bis-1,8-naphthalimide dyes and some ?-acceptors: a spectroscopic study in solution and solid states.  

PubMed

Given the great importance of the various uses of 1,8-naphthalimides in the trends of biology, medicine and industry, the current study focused on extending the scope of these dyes by introducing some of their charge-transfer (CT) complexes. For this purpose, two highly fluorescent bis-1,8-naphthalimide dyes and their complexes with some ?-acceptors have been synthesized and characterized spectroscopically. The ?-acceptors include picric acid (PA), chloranilic acid (CLA), tetracyanoquinodimethane (TCNQ) and dichlorodicyanobenzoquinone (DDQ). The molecular structure, spectroscopic and fluorescence properties as well as the binding modes were deduced from IR, UV-vis and (1)H NMR spectral studies. The binding ratio of complexation was determined to be 1:1 according to the elemental analyses and photometric titrations. It has been found that the order of acceptance ability for the different acceptors is TCNQ>DDQ>CLA>PA. The photostability of 1,8-naphthalimide dye as a donor and its charge-transfer complex doped in polymethyl methacrylate/PMMA were exposed to UV-Vis radiation and the change in the absorption spectra was achieved at different times during irradiation period. PMID:25022501

Refat, Moamen S; Ismail, Lamia A; Adam, Abdel Majid A

2015-01-01

317

Shedding light on the photostability of two intermolecular charge-transfer complexes between highly fluorescent bis-1,8-naphthalimide dyes and some ?-acceptors: A spectroscopic study in solution and solid states  

NASA Astrophysics Data System (ADS)

Given the great importance of the various uses of 1,8-naphthalimides in the trends of biology, medicine and industry, the current study focused on extending the scope of these dyes by introducing some of their charge-transfer (CT) complexes. For this purpose, two highly fluorescent bis-1,8-naphthalimide dyes and their complexes with some ?-acceptors have been synthesized and characterized spectroscopically. The ?-acceptors include picric acid (PA), chloranilic acid (CLA), tetracyanoquinodimethane (TCNQ) and dichlorodicyanobenzoquinone (DDQ). The molecular structure, spectroscopic and fluorescence properties as well as the binding modes were deduced from IR, UV-vis and 1H NMR spectral studies. The binding ratio of complexation was determined to be 1:1 according to the elemental analyses and photometric titrations. It has been found that the order of acceptance ability for the different acceptors is TCNQ > DDQ > CLA > PA. The photostability of 1,8-naphthalimide dye as a donor and its charge-transfer complex doped in polymethyl methacrylate/PMMA were exposed to UV-Vis radiation and the change in the absorption spectra was achieved at different times during irradiation period.

Refat, Moamen S.; Ismail, Lamia A.; Adam, Abdel Majid A.

2015-01-01

318

Loading and release of fluorescent dye from layer-by-layer film-coated magnetic particles in response to hydrogen peroxide.  

PubMed

Polymer-coated magnetic particles (MPs) were prepared to study the binding of fluorescence dye on the surface and its H2O2-induced release. For this goal, multilayer films were prepared by layer-by-layer deposition of shikimic acid-appended poly(allylamine hydrochloride) (SA-PAH) and poly(styrenesulfonate) (PSS) on the surface of MPs. 3-(Dansylamino)phenylboronic acid (DPBA) was loaded on the MPs through boronate ester bonding between SA-PAH and DPBA. DPBA was released from the MPs in response to H2O2 as a result of breakage of the boronate ester bond by an oxidative reaction with H2O2. DPBA release was dependent on the H2O2 concentration. For example, 65% and 93% of the DPBA was released from (SA-PAH/PSS)4SA-PAH film-coated MPs in 30min after the addition of 0.1 and 0.5mM H2O2, respectively. In addition, the multilayer film-coated MPs were further modified by using glucose oxidase (GOx) to develop glucose-induced release systems. GOx-modified MPs released DPBA in response to 0.1mM d-glucose as a result of H2O2 generation through a GOx-catalyzed oxidation reaction of d-glucose. The results suggest a potential use of the multilayer film-coated MPs in the development of H2O2- and/or glucose-sensitive drug delivery systems. PMID:25084230

Sato, Katsuhiko; Abe, Eiichi; Takahashi, Mao; Anzai, Jun-ichi

2014-10-15

319

Deep in vivo two-photon imaging of blood vessels with a new dye encapsulated in pluronic nanomicelles  

NASA Astrophysics Data System (ADS)

Our purpose is to test if Pluronic® fluorescent nanomicelles can be used for in vivo two-photon imaging of both the normal and the tumor vasculature. The nanomicelles were obtained after encapsulating a hydrophobic two-photon dye: di-stryl benzene derivative, in Pluronic block copolymers. Their performance with respect to imaging depth, blood plasma staining, and diffusion across the tumor vascular endothelium is compared to a classic blood pool dye Rhodamin B dextran (70 kDa) using two-photon microscopy. Pluronic nanomicelles show, like Rhodamin B dextran, a homogeneous blood plasma staining for at least 1 h after intravenous injection. Their two-photon imaging depth is similar in normal mouse brain, using 10 times less injected mass. In contrast with Rhodamin B dextran, no extravasation is observed in leaky tumor vessels due to their large size: 20-100 nm. In conclusion, Pluronic nanomicelles can be used as a blood pool dye, even in leaky tumor vessels. The use of Pluronic block copolymers is a valuable approach for encapsulating two-photon fluorescent dyes that are hydrophobic and not suitable for intravenous injection.

Maurin, Mathieu; Stéphan, Olivier; Vial, Jean-Claude; Marder, Seth R.; van der Sanden, Boudewijn

2011-03-01

320

Quick Stain Removal Guide  

E-print Network

. Over time, soil can build up on some fabrics, espe- cially polyesters. A pretreatment product may actually super-clean an area, and may resemble a bleached spot. You can usually correct this by treating the entire garment with a prespotter... or presoak and rewashing with extra detergent. Treat the stain quickly. Time and heat exposure make removing stains harder. Use a blotting motion. Work from the inside of the garment or back of the stain to force the stain out rather than into the fabric...

Brown, Pamela J.

1998-07-29

321

Anthralin stain removal.  

PubMed

Results of an anthralin stain removal study on white 65% polyester/35% cotton, white 100% polyester, white 100% cotton, a white shower curtain, white tile with crevice, and white ceramic shower tile are reported. An optimum stain removal technic was developed by using a 10-minute soak in full-strength chlorine bleach (Good Measure or Clorox) followed by a water rinse and air drying. This technic completely removed all stains of 24-hour duration from the test fabrics. The stain removal test on shower curtains, floor tiles, and ceramic shower tiles was also discussed. PMID:2431014

Wang, J C; Krazmien, R J; Dahlheim, C E; Patel, B

1986-11-01

322

Staining of intracellular deposits of uranium in cultured murine macrophages.  

PubMed

In our studies of the health effects of internalized depleted uranium, we developed a simple and rapid light microscopic method to stain specifically intracellular uranium deposits. Using J774 cells, a mouse macrophage line, treated with uranyl nitrate and the pyridylazo dye 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol, uranium uptake by the cells was followed. Specificity of the stain for uranium was accomplished by using masking agents to prevent the interaction of the stain with other metals. Prestaining wash consisting of a mixture of sodium citrate and ethylenediaminetetraacetic acid eliminated staining of metals other than uranium. The staining solution consisted of the pyridylazo dye in borate buffer along with a quaternary ammonium salt, ethylhexadecyldimethylammonium bromide, and the aforementioned sodium citrate/ethylenediaminetetraacetic acid mixture. The buffer was essential for maintaining the pH within the optimum range of 8 to 12, and the quaternary ammonium salt prevented precipitation of the dye. Staining was conducted at room temperature and was complete in 30 min. Staining intensity correlated with both uranyl nitrate concentration and incubation time. Our method provides a simple procedure for detecting intracellular uranium deposits in macrophages. PMID:11871745

Kalinich, J F; McClain, D E

2001-01-01

323

Near-infrared fluorescence imaging of experimentally collagen-induced arthritis in rats using the nonspecific dye tetrasulfocyanine in comparison with gadolinium-based contrast-enhanced magnetic resonance imaging, histology, and clinical score  

NASA Astrophysics Data System (ADS)

Using 15 rats with collagen-induced arthritis (30 joints) and 7 control rats (14 joints), we correlated the intensity of near-infrared fluorescence (NIRF) of the nonspecific dye tetrasulfocyanine (TSC) with magnetic resonance imaging (MRI), histopathology, and clinical score. Fluorescence images were obtained in reflection geometry using a NIRF camera system. Normalized fluorescence intensity (INF) was determined after intravenous dye administration on different time points up to 120 min. Contrast-enhanced MRI using gadodiamide was performed after NIRF imaging. Analyses were performed in a blinded fashion. Histopathological and clinical scores were determined for each ankle joint. INF of moderate and high-grade arthritic joints were significantly higher (p<0.005) than the values of control and low-grade arthritic joints between 5 and 30 min after TSC-injection. This result correlated well with post-contrast MRI signal intensities at about 5 min after gadodiamide administration. Furthermore, INF and signal increase on contrast-enhanced MRI showed high correlation with clinical and histopathological scores. Sensitivities and specificities for detection of moderate and high-grade arthritic joints were slightly lower for NIRF imaging (89%/81%) than for MRI (100%/91%). NIRF imaging using TSC, which is characterized by slower plasma clearance compared to indocyanine green (ICG), has the potential to improve monitoring of inflamed joints.

Gemeinhardt, Ines; Puls, Dorothee; Gemeinhardt, Ole; Taupitz, Matthias; Wagner, Susanne; Schnorr, Beatrix; Licha, Kai; Schirner, Michael; Ebert, Bernd; Petzelt, Diethard; Macdonald, Rainer; Schnorr, Jörg

2012-10-01

324

Solid acid catalysts: Stain and shine  

NASA Astrophysics Data System (ADS)

Catalyst particles for fluid catalytic cracking are vital for the oil-refinery industry, but their activity is hard to diagnose because of their inter- and intra-particle structural inhomogeneity. With fluorescence confocal microscopy and selective staining, one can now pinpoint the catalytic activity within single catalyst particles from an industrial reactor.

Chen, Peng

2011-11-01

325

Preparation of 6-hydroxyindolines and their use for preparation of novel laser dyes  

DOEpatents

A novel method is described for the synthesis of 6-hydroxyindolines and new fluorescent dyes produced therefrom, which dyes are ring-constrained indoline-based rhodamine class dyes. These dyes have absorption and emission spectra which make them particularly useful in certain dye laser applications.

Field, G.F.; Hammond, P.R.

1993-10-26

326

Preparation of 6-hydroxyindolines and their use for preparation of novel laser dyes  

DOEpatents

A novel method for the synthesis of 6-hydroxyindolines and new fluorescent dyes produced therefrom, which dyes are ring-constrained indoline-based rhodamine class dyes. These dyes have absorption and emission spectra which make them particularly useful in certain dye laser applications.

Field, George F. (Danville, CA); Hammond, Peter R. (Livermore, CA)

1993-01-01

327

Anaphylaxis to annatto dye: a case report.  

PubMed

Annatto dye is an orange-yellow food coloring extracted from the seeds of the tree Bixa orellana. It is commonly used in cheeses, snack foods, beverages, and cereals. Previously reported adverse reactions associated with annatto dye have included urticaria and angioedema. We present a patient who developed urticaria, angioedema, and severe hypotension within 20 minutes following ingestion of milk and Fiber One cereal, which contained annatto dye. Subsequent skin tests to milk, wheat, and corn were negative. The patient had a strong positive skin test to annatto dye, while controls had no response. The nondialyzable fraction of annatto dye on SDS-PAGE demonstrated two protein staining bands in the range of 50 kD. Immunoblotting demonstrated patient IgE-specific for one of these bands, while controls showed no binding. Annatto dye may contain contaminating or residual seed proteins to which our patient developed IgE hypersensitivity. Annatto dye is a potential rare cause of anaphylaxis. PMID:1994783

Nish, W A; Whisman, B A; Goetz, D W; Ramirez, D A

1991-02-01

328

Quinone-methide species, a gateway to functional molecular systems: from self-immolative dendrimers to long-wavelength fluorescent dyes.  

PubMed

Over the last 30 years, the quinone-methide elimination has served as a valuable tool for achieving various important molecular functions. Molecular adaptors based on quinone-methide or aza-quinone-methide reactivity have been designed, synthesized, and used in diagnostic probes, molecular amplifiers, drug delivery systems, and self-immolative dendritic/polymeric molecular systems. These unique adaptors function as stable spacers between an enzyme- or reagent-responsive group and a reporter moiety and can undergo 1,4-, 1,6-, or 1,8-type elimination reactions upon cleavage of the triggering group. Such reactivity results in the release of the reporter group through formation of a quinone-methide species. This type of elimination was applied to design distinct molecular adaptors capable of multiple quinone-methide eliminations. Using this chemistry, we have developed unique molecular structures that are known today as self-immolative dendrimers. These dendrimers disassemble upon a single triggering event in a domino-like manner from the focal point to their periphery with the consequent release of multiple end-groups. Such molecular structures are used in self-immolative dendritic prodrugs and in diagnostic probes to obtain a significant amplification effect. To further enhance amplification, we have developed the dendritic chain reaction, which uses simple molecules to achieve functionality of high-generation virtual self-immolative dendrimers. In addition, we harnessed the quinone-methide elimination reactivity to design polymers that disassemble from head-to-tail initiated by an analyte-responsive event. Following this example, other chemical reactivities were demonstrated by scientists to design such polymeric molecules. In a manner analogous to the quinone-methide elimination, electron rearrangement can lead to formation of conjugated quinone-methide-type dyes with long-wavelength emission of fluorescence. We have recently applied an intramolecular charge transfer to form a unique kind of quinone-methide type derivative based on a donor-two-acceptors molecular structure. This intramolecular charge transfer produces a new fluorochrome with an extended conjugation of ?-electron system that is used for the design of long-wavelength fluorogenic probes with a turn-ON option. The rapidly expanding use of quinone-methide species, reflected in the increased number of examples reported in the literature, indicates the importance of this tool in chemistry. These species provide a useful gateway to functional molecular structures with distinct reactivities and spectroscopic characteristics. PMID:25181456

Gnaim, Samer; Shabat, Doron

2014-10-21

329

Identification of small-scale low and high permeability layers using single well forced-gradient tracer tests: Fluorescent dye imaging and modelling at the laboratory-scale  

NASA Astrophysics Data System (ADS)

Heterogeneity in aquifer permeability, which creates paths of varying mass flux and spatially complex contaminant plumes, can complicate the interpretation of contaminant fate and transport in groundwater. Identifying the location of high mass flux paths is critical for the reliable estimation of solute transport parameters and design of groundwater remediation schemes. Dipole flow tracer tests (DFTTs) and push-pull tests (PPTs) are single well forced-gradient tests which have been used at field-scale to estimate aquifer hydraulic and transport properties. In this study, the potential for PPTs and DFTTs to resolve the location of layered high- and low-permeability layers in granular porous media was investigated with a pseudo 2-D bench-scale aquifer model. Finite element fate and transport modelling was also undertaken to identify appropriate set-ups for in situ tests to determine the type, magnitude, location and extent of such layered permeability contrasts at the field-scale. The characteristics of flow patterns created during experiments were evaluated using fluorescent dye imaging and compared with the breakthrough behaviour of an inorganic conservative tracer. The experimental results show that tracer breakthrough during PPTs is not sensitive to minor permeability contrasts for conditions where there is no hydraulic gradient. In contrast, DFTTs are sensitive to the type and location of permeability contrasts in the host media and could potentially be used to establish the presence and location of high or low mass flux paths. Numerical modelling shows that the tracer peak breakthrough time and concentration in a DFTT is sensitive to the magnitude of the permeability contrast (defined as the permeability of the layer over the permeability of the bulk media) between values of 0.01-20. DFTTs are shown to be more sensitive to deducing variations in the contrast, location and size of aquifer layered permeability contrasts when a shorter central packer is used. However, larger packer sizes are more likely to be practical for field-scale applications, with fewer tests required to characterise a given aquifer section. The sensitivity of DFTTs to identify layered permeability contrasts was not affected by test flow rate.

Barns, Gareth L.; Thornton, Steven F.; Wilson, Ryan D.

2015-01-01

330

Application of fluorescent indicators to analyse intracellular calcium and morphology in filamentous fungi.  

PubMed

A novel staining and quantification method to investigate changes in intracellular calcium levels [Ca(2+)](i) and morphology in filamentous fungus is presented. Using a simple protocol, two fluorescent dyes, Fluo-4-AM and Cell trace calcein red-orange-AM were loaded into the filamentous fungus Penicillium chrysogenum. The present study investigates the applicability of using Ca(2+)-sensitive dye to quantify and image [Ca(2+)](i) in P. chrysogenum cultures chosen for its potential as an experimental system to study Ca(2+) signalling in elicited cultures. The dye loading was optimised and investigated at different pH loading conditions. It was observed that the fluorophore was taken up throughout the hyphae, retaining cell membrane integrity and no dye compartmentalisation within organelles was observed. From the fluorescent plate-reader studies a significant rise (p<0.001) in the relative fluorescence levels corresponding to [Ca(2+)](i) levels in the hyphae was observed when challenged with an elicitor (mannan oligosaccharide, 150mgL(-1)) which was dependent upon extracellular calcium. Concurrently a novel application of dye-loaded hyphae for morphological analysis was also examined using the imaging software Filament Tracer (Bitplane). Essential quantitative mycelial information including the length and diameter of the segments and number of branch points was obtained using this application based on the three-dimensional data. PMID:21530914

Nair, Rakesh; Raina, Sheetal; Keshavarz, Tajalli; Kerrigan, Mark J P

2011-01-01

331

Effect of acidic pH on flow cytometric detection of bacteria stained with SYBR Green I and their distinction from background  

NASA Astrophysics Data System (ADS)

Unspecific background caused by biotic or abiotic particles, cellular debris, or autofluorescence is a well-known interfering parameter when applying flow cytometry to the detection of microorganisms in combination with fluorescent dyes. We present here an attempt to suppress the background signal intensity and thus to improve the detection of microorganisms using the nucleic acid stain SYBR® Green I. It has been observed that the fluorescent signals from SYBR Green I are greatly reduced at acidic pH. When lowering the pH of pre-stained samples directly prior to flow cytometric analysis, we hypothesized that the signals from particles and cells with membrane damage might therefore be reduced. Signals from intact cells, temporarily maintaining a neutral cytosolic pH, should not be affected. We show here that this principle holds true for lowering background interference, whereas the signals of membrane-compromised dead cells are only affected weakly. Signals from intact live cells at low pH were mostly comparable to signals without acidification. Although this study was solely performed with SYBR® Green I, the principle of low pH flow cytometry (low pH-FCM) might hold promise when analyzing complex matrices with an abundance of non-cellular matter, especially when expanded to non-DNA binding dyes with a stronger pH dependence of fluorescence than SYBR Green I and a higher pKa value.

Baldock, Daniel; Nebe-von-Caron, Gerhard; Bongaerts, Roy; Nocker, Andreas

2013-12-01

332

The reaction between phosphatidylethanolamines and HOCl investigated by TLC: fading of the dye primuline is induced by dichloramines.  

PubMed

Phosphatidylethanolamines (PEs) and phosphatidylglycerols (PGs) are abundant lipid constituents of the membranes of Escherichia coli. The reaction between these lipids and hypochlorous acid (HOCl), an important constituent of disinfectants, was investigated by combined thin-layer chromatography (TLC), mass spectrometry (MS), UV and fluorescence spectroscopy. Primuline is a common dye in lipid research that binds non-covalently to lipids and allows, thus, the direct evaluation of TLC plates by MS. However, primuline staining of the products between PE and HOCl is accompanied by fading of the dye. This only holds if acidic but not alkaline conditions are applied. Using a combination of TLC, UV and fluorescence spectroscopy, it will be shown that dichloramines of PE are responsible for the observed primuline fading. Since dichloramines are slowly converted under alkaline conditions into the nitriles that lack the characteristic UV properties of dichloramines, fading is not observed under alkaline conditions. PMID:18440878

Richter, Grit; Schober, Celestina; Süss, Rosmarie; Fuchs, Beate; Müller, Matthias; Schiller, Jürgen

2008-05-15

333

Apparatus Would Stain Microscope Slides  

NASA Technical Reports Server (NTRS)

Proposed apparatus meters specific amounts of fluid out of containers at specific times to stain microscope slides. Intended specifically for semiautomated staining of microbiological and hematological samples in microgravity, leakproof apparatus used in other environments in which technicians have little time to allocate to staining procedures and/or exposure to toxic staining agents or to micro-organisms to be stained hazardous. Apparatus adapted to perform almost any staining procedure and accommodates multiple staining reagents, useful for small or remote clinical laboratories.

Breeding, James D.

1993-01-01

334

Port-Wine Stains  

MedlinePLUS

... their own, they can be treated. In fact, laser therapies can make many port-wine stains much ... mark might be. The good news is that lasers (highly concentrated light energy) can make many kids' ...

335

Fluorometric procedures for dye tracing  

USGS Publications Warehouse

This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The advantages of dye tracing are (1) low detection and measurement limits and (2) simplicity and accuracy in measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section on aerial photography is included because of its possible use to supplement ground-level fluorometry.

Wilson, James F.

1968-01-01

336

Fluorometric procedures for dye tracing  

USGS Publications Warehouse

This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The outstanding characteristics of dye tracing are: (1) the low detection and measurement limits, and (2) the simplicity and accuracy of measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a general guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section is included on aerial photography because of its possible use to supplement ground-level fluorometry. (USGS)

Wilson, James E., Jr.; Cobb, E.D.; Kilpatrick, F.A.

1984-01-01

337

Fluorometric procedures for dye tracing  

USGS Publications Warehouse

This manual describes the current fluorometric procedures used by the U.S. Geological Survey in dye tracer studies such as time of travel, dispersion, reaeration, and dilution-type discharge measurements. The advantages of dye tracing are (1) low detection and measurement limits and (2) simplicity and accuracy in measuring dye tracer concentrations using fluorometric techniques. The manual contains necessary background information about fluorescence, dyes, and fluorometers and a description of fluorometric operation and calibration procedures as a guide for laboratory and field use. The background information should be useful to anyone wishing to experiment with dyes, fluorometer components, or procedures different from those described. In addition, a brief section on aerial photography is included because of its possible use to supplement ground-level fluorometry.

Wilson, James F.; Cobb, Ernest D.; Kilpatrick, F.A.

1986-01-01

338

A low-toxic artificial fluorescent glycoprotein can serve as an efficient cytoplasmic labeling in living cell.  

PubMed

To maintain the virtue of good optical property and discard the dross of conventional fluorescent staining dyes, we provide a strategy for designing new fluorescent scaffolds. In this study, a novel fluorescent labeling glycoprotein (chitosan-poly-l-cysteine, CPC) was synthesized through graft copolymerization. CPC gives emission peak at 465-470nm when excited at 386nm. The submicro-scale CPC microspheres could be localized and persisted specifically in the cytoplasm of living cells, with strong blue fluorescence. Moreover, CPC was highly resistant to photo bleaching, the fluorescence was remained stable for up to 72h as the cells grew and developed. The glycoprotein CPC was bio-compatible and in zero grade cytotoxicity as quantified by MTT assay. The fluorescent labeling process with our newly designed glycoprotein CPC is exceptionally efficient. PMID:25498627

Si, Jiangju; Liang, Dawei; Kong, Dan; Wu, Sufang; Yuan, Lan; Xiang, Yan; Jiang, Lei

2015-03-01

339

Automated four color CD4/CD8 analysis of leukocytes by scanning fluorescence microscopy using Quantum dots  

NASA Astrophysics Data System (ADS)

Scanning Fluorescence Microscope (SFM) is a new technique for automated motorized microscopes to measure multiple fluorochrome labeled cells (Bocsi et al. Cytometry 2004, 61A:1). The ratio of CD4+/CD8+ cells is an important in immune diagnostics in immunodeficiency and HIV. Therefor a four-color staining protocol (DNA, CD3, CD4 and CD8) for automated SFM analysis of lymphocytes was developed. EDTA uncoagulated blood was stained with organic and inorganic (Quantum dots) fluorochromes in different combinations. Aliquots of samples were measured by Flow Cytometry (FCM) and SFM. By SFM specimens were scanned and digitized using four fluorescence filter sets. Automated cell detection (based on Hoechst 33342 fluorescence), CD3, CD4 and CD8 detection were performed, CD4/CD8 ratio was calculated. Fluorescence signals were well separable on SFM and FCM. Passing and Bablok regression of all CD4/CD8 ratios obtained by FCM and SFM (F(X)=0.0577+0.9378x) are in the 95% confidence interval. Cusum test did not show significant deviation from linearity (P>0.10). This comparison indicates that there is no systemic bias between the two different methods. In SFM analyses the inorganic Quantum dot staining was very stable in PBS in contrast to the organic fluorescent dyes, but bleached shortly after mounting with antioxidant and free radical scavenger mounting media. This shows the difficulty of combinations of organic dyes and Quantum dots. Slide based multi-fluorescence labeling system and automated SFM are applicable tools for the CD4/CD8 ratio determination in peripheral blood samples. Quantum Dots are stable inorganic fluorescence labels that may be used as reliable high resolution dyes for cell labeling.

Bocsi, Jozsef; Mittag, Anja; Varga, Viktor S.; Molnar, Bela; Tulassay, Zsolt; Sack, Ulrich; Lenz, Dominik; Tarnok, Attila

2006-02-01

340

[Vital dyes in chromovitrectomy].  

PubMed

The aim of this article is to present the current data with regard to the application of vital dyes during vitreoretinal surgery, 'chromovitrectomy', as well as to overview the current literature regarding the properties of dyes, techniques of application, indications and complications in chromovitrectomy. A large body of published research has recently addressed the toxicity profile of indocyanine green for chromovitrectomy. Experimental data demonstrate dose-dependent toxicity of indocyanine green to various retinal cells. Newer generation vital dyes for chromovitrectomy include trypan blue, patent blue, triamcinolone acetonide, infracyanine green, sodium fluorescein, bromophenol blue, fluorometholone acetate and brilliant blue. Novel instruments may enable a selective painting of preretinal tissues during chromovitrectomy. This review suggests that the field of chromovitrectomy represents an expanding area of research. The first line agents for internal limiting membrane staining in chromovitrectomy are indocyanine green, infracyanine green, and brilliant blue. Patent blue, bromophenol blue and trypan blue arose as outstanding biostains for visualization of epiretinal membranes. Novel dyes available for chromovitrectomy deserve further investigation. PMID:20098913

Dib, Eduardo; Rodrigues, Eduardo Büchelle; Maia, Maurício; Meyer, Carsten H; Penha, Fernando Marcondes; Furlani, Bruno de Albuquerque; Costa, Elaine de Paula Fiod; Farah, Michel Eid

2009-01-01

341

Time-resolved multicolor two-photon excitation fluorescence microscopy of cells and tissues  

NASA Astrophysics Data System (ADS)

Multilabeling which maps the distribution of different targets is an indispensable technique in many biochemical and biophysical studies. Two-photon excitation fluorescence (TPEF) microscopy of endogenous fluorophores combining with conventional fluorescence labeling techniques such as genetically encoded fluorescent protein (FP) and fluorescent dyes staining could be a powerful tool for imaging living cells. However, the challenge is that the excitation and emission wavelength of these endogenous fluorophores and fluorescent labels are very different. A multi-color ultrafast source is required for the excitation of multiple fluorescence molecules. In this study, we developed a two-photon imaging system with excitations from the pump femtosecond laser and the selected supercontinuum generated from a photonic crystal fiber (PCF). Multiple endogenous fluorophores, fluorescent proteins and fluorescent dyes were excited in their optimal wavelengths simultaneously. A time- and spectral-resolved detection system was used to record the TPEF signals. This detection technique separated the TPEF signals from multiple sources in time and wavelength domains. Cellular organelles such as nucleus, mitochondria, microtubule and endoplasmic reticulum, were clearly revealed in the TPEF images. The simultaneous imaging of multiple fluorophores of cells will greatly aid the study of sub-cellular compartments and protein localization.

Zheng, Wei

2014-11-01

342

Identification of senescence and death in Emiliania huxleyi and Thalassiosira pseudonana: Cell staining, chlorophyll alterations, and  

E-print Network

permeability, hydrolytic enzyme, and caspase-like activities using fluorescent cell stains to document diacetate (CMFDA; hydrolytic activity), whereas E. huxleyi developed two distinct CMFDA populations

Berges, John A.

343

Ruby Protein Stains Instruction Manual  

E-print Network

SYPRO® Ruby Protein Stains Instruction Manual SYPRO® Ruby protein gel stain Catalog Numbers 170 ......................................................................................7 2.2 Visualizing and Imaging a Blot .................................................9 PART 4

Lebendiker, Mario

344

[Novelty of vital dyes in ophthalmic surgery].  

PubMed

The aim of this paper is to present recent developments in the area of novelty of vital dyes in intraocular surgery. The authors present the advantages and disadvantages of several vital dyes currently used in ophthalmic surgery. Vital dyes are used to allow better intraoperative visualization of both the anterior and posterior segments. Indocyanine green and trypan blue are the most frequently used and the most efficacious dyes for staining the important anatomic areas but often are associated with significant side effects. These dyes are used in cataract and vitreo-retinal surgery. Other dyes including rhodamine 6G, E68, bromophenol blue, light green and Chicago blue are still under preclinical assessment. PMID:20825072

Rejdak, Robert; Oleszczuk, Agnieszka; Ma?kowska, Anna; Kiczy?ska, Magdalena; Zagórski, Zbigniew; Zarnowski, Tomasz

2010-01-01

345

Reduced Fluorescence Quenching of Cyclodextrin-Acetylene Dye Rotaxanes Jong S. Park, James N. Wilson, Kenneth I. Hardcastle, Uwe H. F. Bunz,*, and  

E-print Network

. Wilson, Kenneth I. Hardcastle,¶ Uwe H. F. Bunz,*, and Mohan Srinivasarao*,,,§ School of Polymer, Textile quantum yield and high oxidation potential due to the electron-withdrawing effect of the -CtC- unit. There is no significant difference in the electronic absorption and emission spectra of free and rotaxanated dyes. However

Srinivasarao, Mohan

346

Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions  

Microsoft Academic Search

The application of the “critical electrolyte concentration” (CEC) concept to the differentiation of acidic glycosaminglycans (mucopolysaccharides) is described. Alcian Blue 8GX stains with increasing selectivity as increasing amounts of magnesium chloride are incorporated into the dye solution. Model experiments with pure polyanions, or artifically carboxylated, phosphorylated and sulphated liver sections, showed that binding of dye to carboxylate or phosphate groups

J. E. Scott; J. Dorling

1965-01-01

347

Stained Glass Glue  

NSDL National Science Digital Library

In this activity on page 6 of the PDF, learners use glue instead of glass to create artwork that can be hung in a window. Discover how the chemicals in various materials mix together to make a colorful, translucent "stained glass" creation.

Society, American C.

2001-01-01

348

Stained-Glass Pastels  

ERIC Educational Resources Information Center

The author has always liked the look of stained-glass windows. Usually the designs are simplified and the shapes are easier for younger students to draw. This technique seemed to be the perfect place for her fifth-graders to try their hand at color mixing. The smaller spaces and simple shapes were just what she needed for this group. Her students…

Laird, Shirley

2009-01-01

349

Fluorescence imaging of microbe-containing particles shot from a two-stage Light-gas gun into an aerogel  

NASA Astrophysics Data System (ADS)

We have proposed an experiment (the Tanpopo mission) to capture microbes on the Japan Experimental Module of the International Space Station. An ultra low-density silica aerogel will be exposed to space for more than 1 year. After retrieving the aerogel, particle tracks and particles found in it will be visualized by fluorescence microscopy after staining it with a DNA-specific fluorescence dye. In preparation for this study, we simulated particle trapping in an aerogel so that methods could be developed to visualize the particles and their tracks. During the Tanpopo mission, particles that have an orbital velocity of ~8 km/s are expected to collide with the aerogel. To simulate these collisions, we shot Deinococcus radiodurans-containing Lucentite particles into the aerogel from a two-stage light-gas gun (acceleration 4.2 km/s). The shapes of the captured particles, and their tracks and entrance holes were recorded with a microscope/camera system for further analysis. The size distribution of the captured particles was smaller than the original distribution, suggesting that the particles had fragmented. We were able to distinguish between microbial DNA and inorganic compounds after staining the aerogel with the DNA-specific fluorescence dye SYBR green I as the fluorescence of the stained DNA and the autofluorescence of the inorganic particles decay at different rates. The developed methods are suitable to determine if microbes exist at the International Space Station altitude.

Kawaguchi, Yuko; Sugino, Tomohiro; Tabata, Makoto; Okudaira, Kyoko; Imai, Eichi; Yano, Hajime; Hasegawa, Sunao; Hashimoto, Hirofumi; Yabuta, Hikaru; Kobayashi, Kensei; Kawai, Hideyuki; Mita, Hajime; Yokobori, Shin-ichi; Yamagishi, Akihiko

2014-02-01

350

Fluorescence imaging of microbe-containing particles shot from a two-stage Light-gas gun into an aerogel.  

PubMed

We have proposed an experiment (the Tanpopo mission) to capture microbes on the Japan Experimental Module of the International Space Station. An ultra low-density silica aerogel will be exposed to space for more than 1 year. After retrieving the aerogel, particle tracks and particles found in it will be visualized by fluorescence microscopy after staining it with a DNA-specific fluorescence dye. In preparation for this study, we simulated particle trapping in an aerogel so that methods could be developed to visualize the particles and their tracks. During the Tanpopo mission, particles that have an orbital velocity of ~8 km/s are expected to collide with the aerogel. To simulate these collisions, we shot Deinococcus radiodurans-containing Lucentite particles into the aerogel from a two-stage light-gas gun (acceleration 4.2 km/s). The shapes of the captured particles, and their tracks and entrance holes were recorded with a microscope/camera system for further analysis. The size distribution of the captured particles was smaller than the original distribution, suggesting that the particles had fragmented. We were able to distinguish between microbial DNA and inorganic compounds after staining the aerogel with the DNA-specific fluorescence dye SYBR green I as the fluorescence of the stained DNA and the autofluorescence of the inorganic particles decay at different rates. The developed methods are suitable to determine if microbes exist at the International Space Station altitude. PMID:25086872

Kawaguchi, Yuko; Sugino, Tomohiro; Tabata, Makoto; Okudaira, Kyoko; Imai, Eichi; Yano, Hajime; Hasegawa, Sunao; Hashimoto, Hirofumi; Yabuta, Hikaru; Kobayashi, Kensei; Kawai, Hideyuki; Mita, Hajime; Yokobori, Shin-ichi; Yamagishi, Akihiko

2014-02-01

351

Fluorescence imaging of microbe-containing particles shot from a two-stage Light-gas gun into an aerogel  

NASA Astrophysics Data System (ADS)

We have proposed an experiment (the Tanpopo mission) to capture microbes on the Japan Experimental Module of the International Space Station. An ultra low-density silica aerogel will be exposed to space for more than 1 year. After retrieving the aerogel, particle tracks and particles found in it will be visualized by fluorescence microscopy after staining it with a DNA-specific fluorescence dye. In preparation for this study, we simulated particle trapping in an aerogel so that methods could be developed to visualize the particles and their tracks. During the Tanpopo mission, particles that have an orbital velocity of ~8 km/s are expected to collide with the aerogel. To simulate these collisions, we shot Deinococcus radiodurans-containing Lucentite particles into the aerogel from a two-stage light-gas gun (acceleration 4.2 km/s). The shapes of the captured particles, and their tracks and entrance holes were recorded with a microscope/camera system for further analysis. The size distribution of the captured particles was smaller than the original distribution, suggesting that the particles had fragmented. We were able to distinguish between microbial DNA and inorganic compounds after staining the aerogel with the DNA-specific fluorescence dye SYBR green I as the fluorescence of the stained DNA and the autofluorescence of the inorganic particles decay at different rates. The developed methods are suitable to determine if microbes exist at the International Space Station altitude.

Kawaguchi, Yuko; Sugino, Tomohiro; Tabata, Makoto; Okudaira, Kyoko; Imai, Eichi; Yano, Hajime; Hasegawa, Sunao; Hashimoto, Hirofumi; Yabuta, Hikaru; Kobayashi, Kensei; Kawai, Hideyuki; Mita, Hajime; Yokobori, Shin-ichi; Yamagishi, Akihiko

2014-08-01

352

Enhancing Analysis of Cells and Proteins by Fluorescence Imaging on Silk-Based Biomaterials: Modulating the Autofluorescence of Silk.  

PubMed

Silk is a versatile and established biomaterial for various tissue engineering purposes. However, it also exhibits strong autofluorescence signals-thereby hindering fluorescence imaging analysis of cells and proteins on silk-derived biomaterials. Sudan Black B (SB) is a lysochrome dye commonly used to stain lipids in histology. It has also been reported to be able to quench autofluorescence of tissues in histology and has been tested on artificial biomedical polymers in recent years. It was hypothesized that SB would exert similar quenching effects on silk, modulating the autofluorescence signals, and thereby enabling improved imaging analysis of cells and molecules of interests. The quenching effect of SB on the intrinsic fluorescence properties of silk and on commercial fluorescent dyes were first investigated in this study. SB was then incorporated into typical fluorescence-based staining protocols to study its effectiveness in improving fluorescence-based imaging of the cells and proteins residing with the silk-based biomaterials. Silk processed into various forms of biomaterials (e.g., films, sponges, fibers, and electrospun mats) was seeded with cells and cultured in vitro. At sacrificial time points, specimens were harvested, fixed, and prepared for fluorescence staining. SB, available commercially as a powder, was dissolved in 70% ethanol (0.3% [w/v]) to form staining solutions. SB treatment was introduced at the last step of typical immunofluorescence staining protocols for 15-120?min. For actin staining protocols by phalloidin toxin, SB staining solutions were added before and after permeabilization with Triton-X for 15-30?min. Results showed that ideal SB treatment duration is about 15?min. Apart from being able to suppress the autofluorescence of silk, this treatment duration was also not too long to adversely affect the fluorescent labeling probes used. The relative improvement brought about by SB treatment was most evident in the blue and green emission wavelengths compared with the red emission wavelength. This study has showed that the use of SB is a cost and time effective approach to enhance fluorescence-based imaging analyses of cell-seeded silk biomaterials, which otherwise would have been hindered by the unmodulated autofluorescence signals. PMID:25050876

Neo, Puay Yong; Tan, Daryl Jian-An; Shi, Pujiang; Toh, Siew Lok; Goh, James Cho-Hong

2014-10-17

353

Direct measurement of the voltage sensitivity of second-harmonic generation from a membrane dye in patch-clamped cells  

NASA Astrophysics Data System (ADS)

We report what is to our knowledge the first optical imaging of voltage-clamped cells by second-harmonic generation. For the membrane-staining styryl dye di-4-ANEPPS, we determined the sensitivity of second-harmonic generation to be 18%/100 mV at an excitation wavelength of 850 nm. This sensitivity is significantly better than the optimal 10%/100 mV under fluorescence and further establishes the importance of second-harmonic generation for the functional imaging of membrane potential in living cells.

Millard, Andrew C.; Jin, Lei; Lewis, Aaron; Loew, Leslie M.

2003-07-01

354

Photobleaching and reorientational dynamics of dyes in a nematic liquid crystal M. Nollmann and D. Shalom  

E-print Network

Received 25 August 1998 The polarized fluorescence of excited dyes in a prototype nematic liquid crystalPhotobleaching and reorientational dynamics of dyes in a nematic liquid crystal M. No¨llmann and D of the liquid crystal, if the dyes are resonantly excited. The fluorescence emission in this latter case is able

Nollmann, Marcelo

355

Suitability of Polymeric Media In Solid State Dye Lasers  

NASA Astrophysics Data System (ADS)

Solid hosts doped with organic dyes are suitable for tunable solid state lasers because of large band width in visible region. Moreover they also overcome the problems of toxicity and limited tunability due to liquid solutions of the dyes. We report fluorescence spectra of different rhodamine dyes in different solid hosts which can be quite helpful in choosing the proper solid host for solid state dye lasers.

Sharma, Amit; Saini, G. S. S.

2011-12-01

356

Photophysics of xanthene dyes in surfactant solution  

NASA Astrophysics Data System (ADS)

The spectral (both absorption and fluorescence) and photoelectrochemical studies of some anionic xanthene dyes namely erythrosin B, rose bengal and eosin have been carried out in micellar solution of cationic cetyl trimethyl ammonium bromide (CTAB), anionic sodium dodecyl sulphate (SDS) and neutral triton X-100 (TX-100). The results show that all these dyes form 1:1 electron-donor-acceptor (EDA) or charge-transfer (CT) complexes with TX-100, which acts as an electron donor. There is no interaction of these dyes with SDS, whereas the interaction with CTAB is mainly electrostatic in nature. In presence of TX-100, these dyes show enhancement of fluorescence intensity with a red shift and develop photovoltage in a photoelectrochemical cell. A good correlation has been found among the photovoltage generation in the systems consisting of these dyes and TX-100, spectral shift due to complex formation and thermodynamic properties of these complexes.

Bhowmik, Benoy B.; Ganguly, Papia

2005-07-01

357

Fluorescent antibody localization of myosin in the cytoplasm, cleavage furrow, and mitotic spindle of human cells  

PubMed Central

We have studied the distribution of myosin molecules in human cells using myosin-specific antibody coupled with fluorescent dyes. Rabbits were immunized with platelet myosin or myosin rod. They produced antisera which precipitated only myosin among all the components in crude platelet extracts. From these antisera we isolated immunoglobulin- G (IgG) and conjugated it with tetramethylrhodamine or fluorescein. We separated IgG with 2-5 fluorochromes per molecule from both under- and over-conjugated IgG by ion exchange chromatography and used it to stain acetone-treated cells. The following controls established the specificity of the staining patterns: (a) staining with labeled preimmune IgG; (b) staining with labeled immune IgG adsorbed with purified myosin; (c) staining with labeled immune IgG mixed with either unlabeled preimmune or immune serum; and (d) staining with labeled antibody purified by affinity chromatography. In blood smears, only the cytoplasm of platelets and leukocytes stained. In spread Enson and HeLa cells, stress fibers stained strongly in closely spaced 0.5 mum spots. The cytoplasm stained uniformly in those cells presumed to be motile before acetone treatment. In dividing HeLa cells there was a high concentration of myosin-specific staining in the vicinity of the contractole ring and in the mitotic spindle, especially the region between the chromosomes and the poles. We detected no staining of erythrocytes, or nuclei of leukocytes and cultured cells, or the surface of platelets and cultured cells. PMID:62755

1976-01-01

358

Determination of cell concentration in a plant cell suspension using a fluorescence microplate reader.  

PubMed

Microscopic counting of plant cells is a very tedious and time-consuming process and is therefore seldom used to evaluate plant cell number on a routine basis. This study describes a fast and simple method to evaluate cell concentration in a plant cell suspension using a fluorescence microplate reader. Eschscholtzia californica cells were fixed in a mix of methanol and acetic acid (3:1) and stained with a fluorescent DNA binding dye (Hoechst 33258). Readings were done in a fluorescence microplate reader at 360/465 nm. Specific binding of the dye to double-stranded DNA was significantly favored over unspecific binding when 1.0 M Tris buffer at pH 7.5 containing 1.0 M NaCl and 75 microg ml(-1) of Hoechst 33258 was used. Fluorescence readings must be done between 4 min and 12 min following the addition of the staining solution to the sample. The microplate counting method provides a convenient, rapid and sensitive procedure for determining the cell concentration in plant cell suspensions. The assay has a linear detection range from 0.2 x 10(6) cells to 10.0 x 10(6) cells per milliliter (actual concentration in the tested cell suspension). The time needed to perform the microplate counting was 10% of that needed for the microscopic enumeration. However, this microplate counting method can only be used on genetically stable cell lines and on asynchronous cell suspensions. PMID:15747158

Lamboursain, Laurence; Jolicoeur, Mario

2005-03-01

359

Dye Painting with Fiber Reactive Dyes  

ERIC Educational Resources Information Center

In her description of how to use dyes directly onto fabrics the author lists materials to be used, directions for mixing dyes, techniques for applying dyes, references for additional reading and sources for dye materials. Preceding the activity with several lessons in design and other textile techniques with the dye process will ensure a…

Benjamin-Murray, Betsy

1977-01-01

360

Multiple equilibria of phenothiazine dyes in aqueous cyclodextrin solutions  

SciTech Connect

A wide variety of applications of the phenothiazine (PN) dyes have been reported, for example, as sensitizers in solar energy conversion. The dimerization and inclusion complexation equilibria of six phenothiazine dyes with cyclodextrins ({alpha}-, {beta}-, and {gamma}-CDs) in aqueous media have been studied using absorption and fluorescence spectroscopy. The PN dyes used in this study are thionine (TH), azure A (AZA), methylene blue (MB), toluidine blue (TB), new methylene blue (NMB), and 1,9-dimethylmethylene blue (DMMB). The dimerization constants (K{sub D}) of the dyes having two methyl substituents at the phenothiazine ring (NMB and DMMB) are much greater than those of other dyes having unsubstituted rings, and the presence of methyl groups on the amine groups affects little the K{sub D} values. The positions of the monomer/dimer equilibria do not change with the presence of {alpha}-CD, while the addition of {beta}-CD suppresses and {gamma}-CD enhances the dimerization of the dyes except DMMB. The equilibrium constants for the inclusion complexation of the dye monomers and dimers with CDs are determined from the analysis of the dependence of the absorption spectra of the dye solutions on the concentrations of the CDs using a multiple-equilibrium scheme. The results indicated that, except DMMB, which has methyl groups at the 1-position of the fused phenothiazine ring, the dye monomers fit better to {beta}-CD and the dimers fit snugly to {gamma}-CD. Fluorescence spectroscopic studies indicate that the dye dimers are not fluorescent and inclusion of the monomer in {beta}-CD results in a 3--5 times enhancement of fluorescence intensity. The determined equilibrium constants of the multiple-equilibrium scheme of the dyes in CD media and fluorescent properties of the dyes can be used to control the dye aggregation and the photophysical and photochemical properties of the phenothiazine dyes for various applications.

Lee, C.; Sung, Y.W.; Park, J.W. [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Chemistry] [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Chemistry

1999-02-04

361

Gram stain of skin lesion  

MedlinePLUS

... of different colored stains is applied to the sample. A laboratory team member examines the stained slide under a microscope, checking for bacteria. The color, size, and shape of the cells help identify the ...

362

Length of stain dosimeter  

NASA Technical Reports Server (NTRS)

Payload customers for the Space Shuttle have recently expressed concerns about the possibility of their payloads at an adjacent pad being contaminated by plume effluents from a shuttle at an active pad as they await launch on an inactive pad. As part of a study to satisfy such concerns a ring of inexpensive dosimeters was deployed around the active pad at the inter-pad distance. However, following a launch, dosimeters cannot be read for several hours after the exposure. As a consequence factors such as different substrates, solvent systems, and possible volatilization of HCl from the badges were studied. This observation led to the length of stain (LOS) dosimeters of this invention. Commercial passive LOS dosimeters are sensitive only to the extent of being capable of sensing 2 ppm to 20 ppm if the exposure is 8 hours. To map and quantitate the HCl generated by Shuttle launches, and in the atmosphere within a radius of 1.5 miles from the active pad, a sensitivity of 2 ppm HCl in the atmospheric gases on an exposure of 5 minutes is required. A passive length of stain dosimeter has been developed having a sensitivity rendering it capable of detecting a gas in a concentration as low as 2 ppm on an exposure of five minutes.

Lueck, Dale E. (inventor)

1994-01-01

363

Development of novel fluorescent probe- protein protected gold nanoclusters for biomedical applications  

NASA Astrophysics Data System (ADS)

This dissertation explores photo-physics, fluorescence polarization properties; resonance energy transfer (RET) probes, and cellular imaging applications for novel fluorescent probe BSA Au clusters By Rayleigh scattering subtraction from extinction spectrum, we found that BSA Au clusters shows peak absorption around 360 nm and shoulder near 500 nm. Peak fluorescence emission lies around 650nm. The fluorescence quantum yield was determined to be 0.06%, while it displayed long fluorescence lifetime of 1.8 mus. BSA Au clusters show stable fluorescence properties and resistance to unfolding and quenching with varying pH, temperature, quencher and denaturant concentration. Moreover, BSA Au clusters shows distinct fluorescence polarization behavior as measured in solvents of different viscosity. The BSA Au cluster, due to long lifetime and high polarization, can potentially be used in studying large macromolecules such as protein complexes with large molecular weight and developing fluorescence polarization immunoassays. BSA Au clusters suffer from several disadvantages such as low quantum efficiency (typically near 6%) and broad emission spectrum (540nm to 800nm). We describe an approach by developing RET probes to enhance the apparent brightness more than 2 fold of BSA Au clusters by linking it with high extinction donor organic dye pacific blue (PB). Moreover, we prepared another conjugate of BSA Au clusters with the near infra-red (NIR) dye Dylight 750 (Dy750), where BSA Au cluster act as a donor to Dy750, showing 46% transfer efficiency to the NIR dye Dy750 with long lifetime component in acceptor decay through RET. Transferring energy from BSA Au cluster to Dy750 will have a RET probe with narrow emission spectrum and long lifetime component which can be explored in imaging applications. Furthermore, herein I describe the use of these long lived BSA Au clusters in cellular and tissue imaging applications. In first approach, I have shown its utility as FLIM probe as well as a time gated intensity imaging probe. In second approach we have shown the one can increase the intensity of long lived BSA Au cluster in cells without increasing the short lived auto-fluorescence background thereby increasing signal to noise ratio at least by 15 times. Furthermore, by applying gated detection strategy to multipulse excitation imaging experiment we increased the signal to noise ratio to 30, a dramatic improvement in contrast for low fluorescence quantum yield dye. In summary, BSA Au clusters can be used as a fluorescent cellular stain advancing the bioimaging technology via their long fluorescence lifetime.

Rostamzadeh Renani, Fatemeh

364

Fluorescence characteristics of 5-carboxytetramethylrhodamine linked covalently to the 5' end of oligonucleotides: multiple conformers of single-stranded and double-stranded dye-DNA complexes.  

PubMed Central

Fluorescence steady-state and lifetime experiments have been carried out on duplex and single-stranded DNA molecules labeled at the 5' ends with 5-carboxytetramethylrhodamine (TMRh). The temperature and ionic strength of the solutions were varied over large ranges. The results reveal at least three well-defined states of the TMRh-DNA molecules for the single-stranded as well as for the double-stranded DNA molecules. Two states are fluorescent, with lifetimes in the range of 0.5-1 ns and 2.5-3 ns. A third state of TMRh-DNA does not fluoresce (a dark species of TMRh-DNA). The distribution of the TMRh-DNA molecules among these three states is strongly temperature and ionic strength dependent. Estimates are made of some reaction parameters of the multistate model. The results are discussed in terms of the photophysics of TMRh, and consequences of the multiple conformers of TMRh-DNA for studies involving fluorescence studies with TMRh-labeled DNA are considered. PMID:8842236

Vámosi, G; Gohlke, C; Clegg, R M

1996-01-01

365

Effect of molecular-level insulation on the performance of a dye-sensitized solar cell: fluorescence studies in solid state.  

PubMed

The performance of a dye-sensitized solar cell (DSSC) that is based on the host-guest encapsulation of 5-[4-diphenylamino)phenyl]thiophene-2-cyanoacrylic acid (L1) inside ?-cyclodextrin hosts has been tested. The formation of the complex in the solid state and when adsorbed on TiO2 was characterized using steady and picosecond time-resolved emission techniques, as well as time dependent DFT calculations. The molecular-level insulation has led to a small enhancement in the energy-conversion performance of the fabricated DSSC with the best results being an increase in the open circuit voltage (Voc) from 0.7 to 0.8 V. The importance of the present investigation lies in the unique spectroscopic characterizations of the examined materials in the solid state. PMID:25398318

Saleh, Na'il; Al-Trawneh, Salah; Al-Dmour, Hmoud; Al-Taweel, Samir; Graham, John P

2015-01-01

366

Environmentally safe removal/disposal of Coomassie Brilliant Blue from gel destain and used gel stain.  

PubMed

Gel destaining following Coomassie Brilliant Blue (CBB) staining involves the use of toxic reagents. Here we demonstrate the efficacy of various paper adsorbents in adsorbing CBB. Kimwipes adsorbed the best, followed by Teri towels, multifold towels, and Whatman numbers 1 and 3 filter papers. Three Kimwipes completely adsorbed the dye released from a CBB-stained mini-gel. Nonradioactive destain solution can, therefore, be recycled for destaining CBB-stained gels. Stain removal with Kimwipes helps in reducing destain use and in reducing organic liquid waste, and it is 7.5-fold cheaper compared with an available method for CBB disposal. Following this, we determined the suitability of this procedure to remove the dye from a used CBB staining solution awaiting proper disposal by our Institutional Safety Office. The dye from a 0.05% CBB staining solution could be removed in 5 to 10 min using 75 Kimwipes. The CBB-adsorbed Kimwipes did not release the stain when squeezed dry even after incubation in various salts over 1week and in water for 5 weeks. The CBB removed allows its easy disposal as solid waste and will not leach out from solid landfills. Thus, stain removal with Kimwipes helps in disposing CBB in an environmentally friendly manner and allows recycling of destaining solution. PMID:20507825

Dorri, Yaser; Kurien, Biji T

2010-09-15

367

Quirks of dye nomenclature. 3. Trypan blue.  

PubMed

Trypan blue is colorant from the 19(th) century that has an association with Africa as a chemotherapeutic agent against protozoan (Trypanosomal) infections, which cause sleeping sickness. The dye still is used for staining biopsies, living cells and organisms, and it also has been used as a colorant for textiles. PMID:24867494

Cooksey, C J

2014-11-01

368

Enhanced Port Wine Stain Lightening Achieved with Combined Treatment of Selective Photothermolysis and Imiquimod  

PubMed Central

Background Pulsed dye laser is the gold standard for treatment of port wine stain birthmarks but multiple treatments are required and complete resolution is often not achieved. Post-treatment vessel recurrence is thought to be a factor that limits efficacy of pulsed dye laser treatment of port wine stains. Imiquimod 5% cream is an immunomodulator with anti-angiogenic effects. Objective To determine if application of imiquimod 5% cream after pulsed dye laser improves treatment outcome. Methods Healthy patients with port wine stains (n = 24) were treated with pulsed dye laser and then randomized to apply post-treatment placebo or imiquimod 5% cream for 8 weeks. Chromameter measurements (CIE L*a*b* colorspace) for 57 port wine stain sites (multiple sites per subject) were taken at baseline and compared with measurements taken 8 weeks post-treatment. The change in a* and ?E were measured to quantify treatment outcome. Results Two subjects developed minor skin irritation. Other adverse effects weren't noted. Average ?a* was 0.43 for pulsed dye laser + placebo sites (n = 25) and 1.27 for pulsed dye laser + imiquimod sites (n = 32) (p value = 0.0294) indicating a greater reduction in erythema with imiquimod. Average ?E was 2.59 for pulsed dye laser + placebo and 4.08 for pulsed dye laser + imiquimod (p value = 0.0363), again indicating a greater color improvement with imiquimod. Limitations Effects were evaluated after a single treatment and duration of effect is unknown. Conclusion Combined selective photothermolysis and anti-angiogenic therapy may enhance port wine stain treatment efficacy. PMID:22244840

Tremaine, Anne Marie; Armstrong, Jennifer; Huang, Yu-Chih; Elkeeb, Leila; Ortiz, Arisa; Harris, Ronald; Choi, Bernard; Kelly, Kristen M.

2012-01-01

369

Measurement of time of travel in streams by dye tracing  

USGS Publications Warehouse

The use of fluorescent dyes and tracing techniques provides a means for measuring the time-of-travel and dispersion characteristics of steady and gradually varied flow in streams. Measurements of the dispersion and concentration of dyes give insight into the behavior of soluble contaminants that may be introduced into a stream. This manual describes methods of measuring time of travel of water and waterborne solutes by dye tracing. The fluorescent dyes, measuring equipment used, and the field and laboratory procedures are also described. Methods of analysis and presentation to illustrate time-oftravel and dispersion characteristics of streams are provided.

Kilpatrick, F.A.; Wilson, James F.

1989-01-01

370

Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes  

PubMed Central

This review presents three examples of using voltage- or calcium-sensitive dyes to image the activity of the brain. Our aim is to discuss the advantages and disadvantages of each method with particular reference to its application to the study of the brainstem. Two of the examples use wide-field (one-photon) imaging; the third uses two-photon scanning microscopy. Because the measurements have limited signal-to-noise ratio, the paper also discusses the methodological aspects that are critical for optimizing the signal. The three examples are the following. (i) An intracellularly injected voltage-sensitive dye was used to monitor membrane potential in the dendrites of neurons in in vitro preparations. These experiments were directed at understanding how individual neurons convert complex synaptic inputs into the output spike train. (ii) An extracellular, bath application of a voltage-sensitive dye was used to monitor population signals from different parts of the dorsal brainstem. We describe recordings made during respiratory activity. The population signals indicated four different regions with distinct activity correlated with inspiration. (iii) Calcium-sensitive dyes can be used to label many individual cells in the mammalian brain. This approach, combined with two-photon microscopy, made it possible to follow the spike activity in an in vitro brainstem preparation during fictive respiratory rhythms. The organic voltage- and ion-sensitive dyes used today indiscriminatively stain all of the cell types in the preparation. A major effort is underway to develop fluorescent protein sensors of activity for selectively staining individual cell types. PMID:19651647

Homma, Ryota; Baker, Bradley J.; Jin, Lei; Garaschuk, Olga; Konnerth, Arthur; Cohen, Lawrence B.; Zecevic, Dejan

2009-01-01

371

Detection of embolization of vessels by a double staining technique.  

PubMed

Air seeding and water vapor embolization of vessels were induced in branches of Acer rubrum by freezing-thawing experiments. A technique has been developed for detecting embolized vessels at the microscopical level. In this technique, one dye is used to label functioning vessels prior to freezing while the second dye is offered after a freezing-thawing cycle at vacuum pressure or at atmospheric pressure in order to reveal water vapor embolization or recovering at 1 bar. Double staining is achieved by differential staining of vessel walls and of their particular contact pits facing paratracheal contact cells. The water potential of the leaves and the rate of uptake of the dye solutions was recorded during the experiments using a Scholander pressure bomb and a potetometer, respectively. While almost 100% of the vessels proved to be functioning in the outer 5 to 7 annual rings, one freezing-thawing cycle inhibited water conduction in up to 82 to 98% of the vessels provided vacuum (0.13 bar) was applied. Rapid recovering however is observed, if the second dye is introduced at atmospheric pressure. PMID:23195303

Sauter, J J

1984-01-01

372

Rapid Microscopy and Use of Vital Dyes: Potential to Determine Viability of Cryptococcus neoformans in the Clinical Laboratory  

PubMed Central

Background Cryptococcus neoformans is the commonest cause of fungal meningitis, with a substantial mortality despite appropriate therapy. Quantitative culture of cryptococci in cerebrospinal fluid (CSF) during antifungal therapy is of prognostic value and has therapeutic implications, but is slow and not practicable in many resource-poor countries. Methods We piloted two rapid techniques for quantifying viable cryptococci using mixtures of live and heat-killed cryptococci cultured in vitro: (i) quantitative microscopy with exclusion staining using trypan blue dye, and (ii) flow cytometry, using the fluorescent dye 2?-7?-Bis-(2-carboxyethyl)-5-(6)-carboxyfluorescein, acetoxymethyl ester (BCECF-AM). Results were compared with standard quantitative cryptococcal cultures. Quantitative microscopy was also performed on cerebrospinal fluid (CSF) samples. Results Both microscopy and flow cytometry distinguished between viable and non-viable cryptococci. Cell counting (on log scale) by microscopy and by quantitative culture were significantly linearly associated (p<0.0001) and Bland-Altman analysis showed a high level of agreement. Proportions of viable cells (on logit scale), as detected by flow cytometry were significantly linearly associated with proportions detected by microscopy (p<0.0001) and Bland-Altman analysis showed a high level of agreement. Conclusions Direct microscopic examination of trypan blue-stained cryptococci and flow-cytometric assessment of BCECF-AM-stained cryptococci were in good agreement with quantitative cultures. These are promising strategies for rapid determination of the viability of cryptococci, and should be investigated in clinical practice. PMID:25625210

McMullan, Brendan J.; Desmarini, Desmarini; Djordjevic, Julianne T.; Chen, Sharon C-A.; Roper, Michael; Sorrell, Tania C.

2015-01-01

373

Phosphomolybdic and Phosphotungstic Acid-Victoria Blue R Stains Two Histochemically Distinct Collagens: Dense Dark Blue and Loose Areolar Pale Gree  

Microsoft Academic Search

Victoria blue stains nuclei and elastic tissue but aqueous mounting media are required since stains are completely washed out by usual alcohol dehydration. Afterchroming with phosphomolybdic (PMo) or phosphotungstic (PW) acid converts the dye to an alcohol insoluble pigment. This is also insoluble in 1 N HCI and in acid alcohol. Premordanting with PMo or PW yields a staining pattern

R. D. LILLIE; C. REYNOLDS; P. PIZZOLATO

374

Rapid in situ assessment of physiological activities in bacterial biofilms using fluorescent probes  

NASA Technical Reports Server (NTRS)

Two rapid in situ enumeration methods using fluorescent probes were used to assess the physiological activities of Klebsiella pneumoniae biofilms on stainless steel. Fluorescent dyes, 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and rhodamine 123 (Rh 123), were chosen to perform this study. CTC is a soluble redox indicator which can be reduced by respiring bacteria to fluorescent CTC-formazan crystals. Rh 123 is incorporated into bacteria with respect to cellular proton motive force. The intracellular accumulation of these fluorescent dyes can be determined using epifluorescence microscopy. The results obtained with these two fluorescent probes in situ were compared to the plate count (PC) and in situ direct viable count (DVC) methods. Viable cell densities within biofilms determined by the three in situ methods were comparable and always showed approximately 2-fold higher values than those obtained with the PC method. As an additional advantage, the results were observed after 2 h, which was shorter than the 4 h incubation time required for the DVC method and 24 h for colony formation. The results indicate that staining with CTC and Rh 123 provides rapid information regarding cell numbers and physiological activities of bacteria within biofilms.

Yu, F. P.; McFeters, G. A.

1994-01-01

375

ORIGINAL PAPER Optimization of the Near-Infrared Fluorescence Labeling  

E-print Network

ORIGINAL PAPER Optimization of the Near-Infrared Fluorescence Labeling for In Vivo Monitoring The objective of this study is to optimize the parameters in labeling near-infrared (NIR)fluorescent dye cypate Near infrared . Fluorescence dye . Protein drugs . Labeling condition . In vivo optical imaging

Larson-Prior, Linda

376

Treatment of port-wine stains: analysis.  

PubMed

Port-wine stains (PWS) are bluish red skin stains that are caused by enlarged, ectatic blood vessels in the dermis. Laser treatment of PWS is analyzed from computation of the spatial distribution of heat production by direct absorption of the laser light and subsequent heat conduction. The absorption and scattering caused by oxyhemoglobin, epidermis, and dermis as a function of wavelength are utilized in this analysis. Ideal treatment is defined as coagulating the ectatic blood vessels without irreversible damage to the epidermis and dermis. The analysis shows that a millisecond pulsed, yellow dye laser at 577 nm (one of the large absorption bands in blood) is the laser of choice to treat PWS, offering as close to the "ideal treatment" as possible. The blue-green argon laser, which is currently the most frequently used laser for this purpose, is strongly recommended with irradiation times in milliseconds. Other lasers that are in clinical use, such as the red ruby and near-infrared Nd-YAG lasers, can provide selective treatment only when the epidermis is cooled concurrently. The CO2 laser, on the other hand, can coagulate the blood vessels only through heat conduction from the hot epidermis; hence, it has neither the treatment selectivity nor any other physical option to force this selectivity. PMID:3452741

van Gemert, M J; Welch, A J

1987-08-01

377

Chromoendoscopy and intravital staining techniques.  

PubMed

Chromoendoscopy and intravital staining techniques are synonymous methods for the endoscopic early detection of malignant changes in the intestinal tract. Endoscopic intravital staining involves the use of absorptive stains (methylene blue and Lugol's solution), contrast stains (indigo carmine) and reactive stains (Congo red). Lugol's iodine solution is used to identify superficial carcinomas in the squamous epithelium of the oesophagus. Methylene blue stains the specialized intestinal epithelium in Barrett's oesophagus and, in addition to this, is helpful in the diagnosis of dysplasia. Intravital staining with indigo carmine contributes to contrasting and accentuating changed mucosal processes. Together with Cresyl violet, contrast staining is particularly important in detecting small, early malignant changes in the colon. The use of chromoendoscopy enables a biopsy diagnosis of superficial dysplastic changes and an accurate delineation of carcinomatous areas. In conjunction with the modern video-endoscopy (high-resolution endoscopy and magnification endoscopy), vital staining forms the diagnostic foundation for the detection of early malignant changes in the gastrointestinal tract. These techniques are therefore prerequisites to local endoscopic tumour therapy (mucosectomy). Despite their increasing acceptance, these methods must prove their diagnostic merit in randomized studies. PMID:11030630

Jung, M; Kiesslich, R

1999-04-01

378

Efficient Blind Spectral Unmixing of Fluorescently Labeled Samples Using Multi-Layer Non-Negative Matrix Factorization  

PubMed Central

The ample variety of labeling dyes and staining methods available in fluorescence microscopy has enabled biologists to advance in the understanding of living organisms at cellular and molecular level. When two or more fluorescent dyes are used in the same preparation, or one dye is used in the presence of autofluorescence, the separation of the fluorescent emissions can become problematic. Various approaches have been recently proposed to solve this problem. Among them, blind non-negative matrix factorization is gaining interest since it requires little assumptions about the spectra and concentration of the fluorochromes. In this paper, we propose a novel algorithm for blind spectral separation that addresses some of the shortcomings of existing solutions: namely, their dependency on the initialization and their slow convergence. We apply this new algorithm to two relevant problems in fluorescence microscopy: autofluorescence elimination and spectral unmixing of multi-labeled samples. Our results show that our new algorithm performs well when compared with the state-of-the-art approaches for a much faster implementation. PMID:24260120

Zudaire, Isabel; Ortiz-de-Solorzano, Carlos

2013-01-01

379

NIR Dyes for Bioimaging Applications  

PubMed Central

Summary of recent advances Fluorescent dyes based on small organic molecules that function in the near infra red (NIR) region are of great current interest in chemical biology. They allow for imaging with minimal autofluorescence from biological samples, reduced light scattering and high tissue penetration. Herein, examples of ongoing NIR fluorophore design strategies as well as their properties and anticipated applications relevant to the bioimaging are presented. PMID:19926332

Escobedo, Jorge O.; Rusin, Oleksandr; Lim, Soojin

2009-01-01

380

Dye laser amplifier  

DOEpatents

An improved dye laser amplifier is disclosed. The efficiency of the dye laser amplifier is increased significantly by increasing the power of a dye beam as it passes from an input window to an output window within the dye chamber, while maintaining the intensity of the dye beam constant. 3 figs.

Moses, E.I.

1992-12-01

381

Biomat flow: fluorescent dye field experiments, pore-scale modeling of flow and transport properties, and field-scale flow models  

NASA Astrophysics Data System (ADS)

Recent studies highlight the important role that the upper litter layer in forest soils (biomat) plays in hillslope and catchment runoff generation. This biomat layer is a very loose material with high porosity and organic content. Direct sampling is usually problematic due to limited layer thickness. Conventional laboratory measurements can mobilize solids or even cause structure failure of the sample thus making measurements unreliable. It is also difficult to assess local variation in soil properties and transition zones using these methods; thus, they may not be applicable to biomat studies. However, if the physics of flow through this layer needs to be quantified and incorporated into a model, a detailed study of hydraulic properties is necessary. Herein we show the significance of biomat flow by staining experiments in the field, study its structure and transition to mineral soil layer using X-ray micro-tomography, assess hydraulic properties and structure differences using a pore-scale modeling approach, and, finally, use conventional variably-saturated flow modeling based on Richards equation to simulate flow in the hillslope. Using staining tracers we show that biomat flow in forested hillslopes can extend long distances (lateral displacement was about 1.2 times larger than for subsurface lateral flow) before infiltration occurs into deeper layers. The three-dimensional structure of an undisturbed sample (4 x 3 x 2.5 cm) of both biomat and deeper consolidated soil was obtained using an X-ray micro-tomography device with a resolution of 15 um. Local hydraulic properties (e.g., permeability and water retention curve) for numerous layers (e.g., transition zones, biomat, mineral soil) were calculated using Stokes flow FDM solution and pore-network modeling. Anisotropy, structure differences, and property fluctuations of different layers were quantified using local porosity analysis and correlation functions. Current results support the hypothesis that small-scale structural differences alone can explain the lateral transport observed in field. Possible water repellent behaviour at the biomat-mineral soil interface may also be contributing to extended surface flow. Based on conventional modeling with HYDRUS-2D we show how pore-scale modelling-derived hydraulic properties can improve large scale simulations and clarify how local heterogeneities in wetting and hydraulic properties affect infiltration into deeper soils and likely magnify preferential flow. This work was partially supported by RFBR grants 12-05-33089, 12-04-32264, 13-04-00409, 13-05-01176 and 12-05-01130.

Gerke, K.; Sidle, R. C.; Mallants, D.; Vasilyev, R.; Karsanina, M.; Skvortsova, E. B.; Korost, D. V.

2013-12-01

382

Biomat flow: fluorescent dye field experiments, pore-scale modeling of flow and transport properties, and field-scale flow models  

NASA Astrophysics Data System (ADS)

Recent studies highlight the important role that the upper litter layer in forest soils (biomat) plays in hillslope and catchment runoff generation. This biomat layer is a very loose material with high porosity and organic content. Direct sampling is usually problematic due to limited layer thickness. Conventional laboratory measurements can mobilize solids or even cause structure failure of the sample thus making measurements unreliable. It is also difficult to assess local variation in soil properties and transition zones using these methods; thus, they may not be applicable to biomat studies. However, if the physics of flow through this layer needs to be quantified and incorporated into a model, a detailed study of hydraulic properties is necessary. Herein we show the significance of biomat flow by staining experiments in the field, study its structure and transition to mineral soil layer using X-ray micro-tomography, assess hydraulic properties and structure differences using a pore-scale modeling approach, and, finally, use conventional variably-saturated flow modeling based on Richards equation to simulate flow in the hillslope. Using staining tracers we show that biomat flow in forested hillslopes can extend long distances (lateral displacement was about 1.2 times larger than for subsurface lateral flow) before infiltration occurs into deeper layers. The three-dimensional structure of an undisturbed sample (4 x 3 x 2.5 cm) of both biomat and deeper consolidated soil was obtained using an X-ray micro-tomography device with a resolution of 15 um. Local hydraulic properties (e.g., permeability and water retention curve) for numerous layers (e.g., transition zones, biomat, mineral soil) were calculated using Stokes flow FDM solution and pore-network modeling. Anisotropy, structure differences, and property fluctuations of different layers were quantified using local porosity analysis and correlation functions. Current results support the hypothesis that small-scale structural differences alone can explain the lateral transport observed in field. Possible water repellent behaviour at the biomat-mineral soil interface may also be contributing to extended surface flow. Based on conventional modeling with HYDRUS-2D we show how pore-scale modelling-derived hydraulic properties can improve large scale simulations and clarify how local heterogeneities in wetting and hydraulic properties affect infiltration into deeper soils and likely magnify preferential flow. This work was partially supported by RFBR grants 12-05-33089, 12-04-32264, 13-04-00409, 13-05-01176 and 12-05-01130.

Gerke, K.; Sidle, R. C.; Mallants, D.; Vasilyev, R.; Karsanina, M.; Skvortsova, E. B.; Korost, D. V.

2011-12-01

383

Novel aminobenzanthrone dyes for amyloid fibril detection  

NASA Astrophysics Data System (ADS)

A series of novel fluorescent aminobenzanthrone dyes have been tested for their ability to identify and characterize the oligomeric and fibrillar aggregates of lysozyme. The parameters of the dye binding to native, oligomeric and fibrillar protein have been calculated from the results of fluorimetric titration. Furthermore, several additional quantities reflecting the preference of the probe to either pre-fibrillar or fibrillar protein aggregates, have been evaluated. Based on the comparative analysis of the recovered parameters, AM4 was recommended for selective detection of protein pre-fibrillar assemblies, while the dyes AM1, AM2, AM3 were selected as the most prospective amyloid tracers.

Vus, Kateryna; Trusova, Valeriya; Gorbenko, Galyna; Kirilova, Elena; Kirilov, Georgiy; Kalnina, Inta; Kinnunen, Paavo

2012-04-01

384

Polypeptide micelles with dual pH activatable dyes for sensing cells and cancer imaging  

NASA Astrophysics Data System (ADS)

pH is an important control parameter for maintenance of cell viability and tissue functions. pH monitoring provides valuable information on cell metabolic processes and the living environment. In this study, we prepared dual pH-sensitive, fluorescent dye-loaded polypeptide nanoparticles (DPNs) for ratiometric sensing of pH changes in living cells. DPNs contain two types of dyes: N-(rhodamine B) lactam cystamine (RBLC), an acid activatable fluorescent dye with increased fluorescence in an acidic environment, and fluorescein isothiocyanate (FITC), a base activatable fluorescent dye with enhanced fluorescence in an alkaline environment. Hence, DPNs exhibited a dual response signal with strong red fluorescence and weak green fluorescence under acidic conditions; in contrast, they showed strong green fluorescence and almost no red fluorescence under alkaline and neutral conditions. The favorable inverse pH responses of the two fluorescent dyes resulted in ratiometric pH determination for DPNs with an optimized pH-sensitive range of pH 4.5-7.5. Quantitative analysis of the intracellular pH of intact MCF-7 cells has been successfully demonstrated with our nanosensor. Moreover, single acid activatable fluorescent dye doped polypeptide nanoparticles that only contained RBLC can distinguish tumor tissue from normal tissue by monitoring the acidic extracellular environment.pH is an important control parameter for maintenance of cell viability and tissue functions. pH monitoring provides valuable information on cell metabolic processes and the living environment. In this study, we prepared dual pH-sensitive, fluorescent dye-loaded polypeptide nanoparticles (DPNs) for ratiometric sensing of pH changes in living cells. DPNs contain two types of dyes: N-(rhodamine B) lactam cystamine (RBLC), an acid activatable fluorescent dye with increased fluorescence in an acidic environment, and fluorescein isothiocyanate (FITC), a base activatable fluorescent dye with enhanced fluorescence in an alkaline environment. Hence, DPNs exhibited a dual response signal with strong red fluorescence and weak green fluorescence under acidic conditions; in contrast, they showed strong green fluorescence and almost no red fluorescence under alkaline and neutral conditions. The favorable inverse pH responses of the two fluorescent dyes resulted in ratiometric pH determination for DPNs with an optimized pH-sensitive range of pH 4.5-7.5. Quantitative analysis of the intracellular pH of intact MCF-7 cells has been successfully demonstrated with our nanosensor. Moreover, single acid activatable fluorescent dye doped polypeptide nanoparticles that only contained RBLC can distinguish tumor tissue from normal tissue by monitoring the acidic extracellular environment. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr00519h

Gong, Ping; Yang, Yueting; Yi, Huqiang; Fang, Shengtao; Zhang, Pengfei; Sheng, Zonghai; Gao, Guanhui; Gao, Duyang; Cai, Lintao

2014-04-01

385

FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells.  

PubMed

FM-dyes are widely used to study endocytosis, vesicle trafficking and organelle organization in living eukaryotic cells. The increasing use of FM-dyes in plant cells has provoked much debate with regard to their suitability as endocytosis markers, which organelles they stain and the precise pathways they follow through the vesicle trafficking network. A primary aim of this article is to assess critically the current status of this debate in plant cells. For this purpose, background information on the important characteristics of the FM-dyes, and of optimal dye concentrations, conditions of dye storage, and staining and imaging protocols, are provided. Particular emphasis is placed on using the FM-dyes in double labelling experiments to identity specific organelles. In this way, staining of the Golgi with FM4-64 has been demonstrated for the first time. PMID:15102063

Bolte, S; Talbot, C; Boutte, Y; Catrice, O; Read, N D; Satiat-Jeunemaitre, B

2004-05-01

386

Identification of an ABCB1 (P-glycoprotein)-positive carfilzomib-resistant myeloma subpopulation by the pluripotent stem cell fluorescent dye CDy1.  

PubMed

Multiple myeloma (MM) is characterized by the malignant expansion of differentiated plasma cells. Although many chemotherapeutic agents display cytotoxic activity toward MM cells, patients inevitably succumb to their disease because the tumor cells become resistant to the anticancer drugs. The cancer stem cell hypothesis postulates that a small subpopulation of chemotherapy-resistant cancer cells is responsible for propagation of the tumor. Herein we report that efflux of the pluripotent stem cell dye CDy1 identifies a subpopulation in MM cell lines characterized by increased expression of P-glycoprotein, a member of the ABC (ATP-binding cassette) superfamily of transporters encoded by ABCB1. We also demonstrate that ABCB1-overexpressing MM cells are resistant to the second-generation proteasome inhibitor carfilzomib that recently received accelerated approval for the treatment of therapy-refractive MM by the U.S. Food and Drug Administration. Moreover, increased resistance to carfilzomib in sensitive MM cells following drug selection was associated with upregulation of ABCB1 cell-surface expression which correlated with increased transporter activity as measured by CDy1 efflux. We further show that chemosensitization of MM cells to carfilzomib could be achieved in vitro by cotreatment with vismodegib, a hedgehog pathway antagonist which is currently in MM clinical trials. CDy1 efflux may therefore be a useful assay to determine whether high expression of ABCB1 is predictive of poor clinical responses in MM patients treated with carfilzomib. Our data also suggest that inclusion of vismodegib might be a potential strategy to reverse ABCB1-mediated drug resistance should it occur. PMID:23475625

Hawley, Teresa S; Riz, Irene; Yang, Wenjing; Wakabayashi, Yoshiyuki; Depalma, Louis; Chang, Young-Tae; Peng, Weiqun; Zhu, Jun; Hawley, Robert G

2013-04-01

387

ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES  

EPA Science Inventory

The U.S. Environmental Protection Agency's Method 1623 is used to detect and quantify the presence of {ital Cryptosporidium} spp. oocysts in water. The protocol consists of concentrating a sample, staining this concentrate with a fluorescent antibody, and examining the sample mi...

388

Molecular cytogenetics using fluorescence in situ hybridization  

SciTech Connect

Fluorescence in situ hybridization (FISH) with chromosome-specific probes enables several new areas of cytogenetic investigation by allowing visual determination of the presence and normality of specific genetic sequences in single metaphase or interphase cells. in this approach, termed molecular cytogenetics, the genetic loci to be analyzed are made microscopically visible in single cells using in situ hybridization with nucleic acid probes specific to these loci. To accomplish this, the DNA in the target cells is made single stranded by thermal denaturation and incubated with single-stranded, chemically modified probe under conditions where the probe will anneal only with DNA sequences to which it has high DNA sequence homology. The bound probe is then made visible by treatment with a fluorescent reagent such as fluorescein that binds to the chemical modification carried by the probe. The DNA to which the probe does not bind is made visible by staining with a dye such as propidium iodide that fluoresces at a wavelength different from that of the reagent used for probe visualization. We show in this report that probes are now available that make this technique useful for biological dosimetry, prenatal diagnosis and cancer biology. 31 refs., 3 figs.

Gray, J.W.; Kuo, Wen-Lin; Lucas, J.; Pinkel, D.; Weier, H-U.; Yu, Loh-Chung.

1990-12-07

389

Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms - which stain is suitable?  

PubMed Central

Background There is confusion over the definition of the term “viability state(s)” of microorganisms. “Viability staining” or “vital staining techniques” are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data. Discussion Many terms describe “vitality states” of microorganisms, however, several of them are misleading. Authors define “viable” as “capable to grow”. Accordingly, staining methods are substitutes, since no staining can prove viability. The reliability of a commercial “viability” staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the “viability” kit are dependent on the stains’ concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique. To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research. Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported. Summary –?The nomenclature regarding “viability” and “vitality” should be used carefully. –?The manual of the commercial “viability” kit itself points out that the kit is not suitable for natural multispecies biofilm research, as supported by an array of literature. –?Results obtained with various stains are influenced by the relationship between bacterial counts and the amount of stain used in the test. Corresponding vitality data are prone to artificial shifting. –?As microbiological parameter the Plating Efficiency should be used for comparison. –?Ethidium Bromide is mutagenic. Researchers should be aware that alternative staining compounds may also be or even are mutagenic. PMID:24410850

2014-01-01

390

Automated Feulgen staining with a temperature-controlled staining machine.  

PubMed

A Shandon Varistain 24-3 staining machine was modified in order to run automated DNA Feulgen staining. Initial studies showed a strict dependence of the staining intensity (integrated optical density [IOD]) on the temperature of the DNA hydrolysis in 4 N HCl: a difference of 0.5 degrees C around the optimum hydrolysis temperature of 27.5 degrees C resulted in IOD differences of up to 7.8% in epithelial cells and up to 12.0% in lymphocytes. A temperature-controlled stainless steel cuvette, covered with a 4 N HCl-resistant material, was developed and integrated into the machine. Temperature measurements were performed at different positions in the cuvette and on glass slides with copper-constantan electrodes fixed on them; no temperature gradient could be detected within the cuvette. The adjusted temperature of 27.5 degrees C remained constant over 24 hours. The coefficient of variation (CV) of the staining intensity in lymphocytes between different areas on the same slide and between different slides of the same staining cycle was less than 0.6%. The CV between different staining cycles was 5.9%. This system for automated Feulgen staining thus gives reproducible and reliable results and may be introduced into routine diagnostic procedures. PMID:2472804

Chatelain, R; Willms, A; Biesterfeld, S; Auffermann, W; Böcking, A

1989-06-01

391

Certain tricyclic and pentacyclic-hetero nitrogen rhodol dyes  

DOEpatents

Novel fluorescent dyes based on the rhodol structure are provided. The new reagents contain functional groups capable of forming a stable fluorescent product with functional groups typically found in biomolecules or polymers including amines, phenols, thiols, acids, aldehydes and ketones. Reactive groups in the rhodol dyes include activated esters, isothiocyanates, amines, hydrazines, halides, acids, azides, maleimides, aldehydes, alcohols, acrylamides and haloacetamides. The products are detected by their absorbance or fluorescence properties. The spectral properties of the fluorescent dyes are sufficiently similar in wavelengths and intensity to fluorescein or rhodamine derivatives as to permit use of the same equipment. The dyes, however, show less spectral sensitivity to pH in the physiological range than does fluorescein, have higher solubility in non-polar solvents and have improved photostability and quantum yields.

Haugland, Richard P. (Eugene, OR); Whitaker, James E. (Eugene, OR)

1993-01-01

392

Double-Staining Method for Differentiation of Morphological Changes and Membrane Integrity of Campylobacter coli Cells  

PubMed Central

We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells. PMID:12324366

Alonso, Jose L.; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A.; Hernández, Javier

2002-01-01

393

Double-staining method for differentiation of morphological changes and membrane integrity of Campylobacter coli cells.  

PubMed

We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells. PMID:12324366

Alonso, Jose L; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A; Hernández, Javier

2002-10-01

394

Verifying Removal Of Red Penetrant Dye From Inspected Welds  

NASA Technical Reports Server (NTRS)

Clean surface assured for more sensitive inspection with fluorescent penetrant dye. Simple procedure devised to ensure visible (red) penetrant dye used to identify flaws in welded surface completely removed from surface. Consists in applying reversible penetrant developer to surface to be inspected.

Torkelson, Jan R.

1996-01-01

395

Just Dyeing to Find Out.  

ERIC Educational Resources Information Center

Presents a multidisciplinary unit on natural dyes designed to take advantage of the natural curiosity of middle school students. Discusses history of dyes, natural dyes, preparation of dyes, and the dyeing process. (JRH)

Monhardt, Becky Meyer

1996-01-01

396

Never say dye  

PubMed Central

Recent years have seen a remarkable increase in the number of publications dealing with the application of epifluorescence microscopy in cell biology. This can be widely attributed to the development of state-of-the-art image processing programs, as well as the development of new reagents/probes, which allow the labeling of most cell structures, organelles and metabolites with high specificity. However, the use of a specific fluorescent dye, 3,3?-dihexyloxacarbocyanine iodide (DiOC6), has been recently revisited and several new application potentials have emerged. The goal of this mini-review is to provide an up-to-date overview of the multiple roles of this multifaceted probe. PMID:22476459

2012-01-01

397

Flow cytometric readout based on Mitotracker Red CMXRos staining of live asexual blood stage malarial parasites reliably assesses antibody dependent cellular inhibition  

PubMed Central

Background Functional in vitro assays could provide insights into the efficacy of malaria vaccine candidates. For estimating the anti-parasite effect induced by a vaccine candidate, an accurate determination of live parasite count is an essential component of most in vitro bioassays. Although traditionally parasites are counted microscopically, a faster, more accurate and less subjective method for counting parasites is desirable. In this study mitochondrial dye (Mitotracker Red CMXRos) was used for obtaining reliable live parasite counts through flow cytometry. Methods Both asynchronous and tightly synchronized asexual blood stage cultures of Plasmodium falciparum were stained with CMXRos and subjected to detection by flow cytometry and fluorescence microscopy. The parasite counts obtained by flow cytometry were compared to standard microscopic counts obtained through examination of Giemsa-stained thin smears. A comparison of the ability of CMXRos to stain live and compromised parasites (induced by either medium starvation or by anti-malarial drug treatment) was carried out. Finally, parasite counts obtained by CMXRos staining through flow cytometry were used to determine specific growth inhibition index (SGI) in an antibody-dependent cellular inhibition (ADCI) assay. Results Mitotracker Red CMXRos can reliably detect live intra-erythrocytic stages of P. falciparum. Comparison between staining of live with compromised parasites shows that CMXRos predominantly stains live parasites with functional mitochondria. Parasite counts obtained by CMXRos staining and flow cytometry were highly reproducible and can reliably determine the ability of IgG from hyper-immune individuals to inhibit parasite growth in presence of monocytes in ADCI assay. Further, a dose-dependent parasite growth inhibitory effect could be detected for both total IgG purified from hyper-immune sera and affinity purified IgGs against the N-terminal non-repeat region of GLURP in ADCI assays coupled with determination of parasite counts through CMXRos staining and flow cytometry. Conclusions A flow cytometry method based on CMXRos staining for detection of live parasite populations has been optimized. This is a rapid and sensitive method with high inter-assay reproducibility which can reliably determine the anti-parasite effect mediated by antibodies in functional in vitro assays such as ADCI assay. PMID:22818754

2012-01-01

398

DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy  

PubMed Central

This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. PMID:24727376

Osipov, Andreyan; Arkhangelskaya, Ekaterina; Vinokurov, Alexei; Smetanina, Nadezhda; Zhavoronkov, Alex; Klokov, Dmitry

2014-01-01

399

Photophysical properties of a new DyLight 594 dye.  

PubMed

We describe spectral properties of novel fluorescence probe DyLight 594. Absorption and fluorescence spectra of this dye are in the region of Alexa 594 fluor spectra. The quantum yield of DyLight 594 in conjugated form to IgG is higher than corresponding quantum yield of Alexa 594 by about 50%. The new DyLight dye also shows slightly longer lifetime and photostability. These favorable properties and high anisotropy value, as well as a high cross-section for two-photon excitation, make this fluorophore attractive as a fluorescence probe in biochemical/biological studies involving fluorescence methods. PMID:19948414

Sarkar, Pabak; Sridharan, Savitha; Luchowski, Rafal; Desai, Surbhi; Dworecki, Boguslawa; Nlend, Marie; Gryczynski, Zygmunt; Gryczynski, Ignacy

2010-01-21

400

Fluorescent dye imaging of the volume sampled by single well forced-gradient tracer tests evaluated in a laboratory-scale aquifer physical model  

NASA Astrophysics Data System (ADS)

This study presents a new method to visualise forced-gradient tracer tests in 2-D using a laboratory-scale aquifer physical model. Experiments were designed to investigate the volume of aquifer sampled in vertical dipole flow tracer tests (DFTT) and push-pull tests (PPT), using a miniature monitoring well and straddle packer arrangement equipped with solute injection and recovery chambers. These tests have previously been used to estimate bulk aquifer hydraulic and transport properties for the evaluation of natural attenuation and other remediation approaches. Experiments were performed in a silica glass bead-filled box, using a fluorescent tracer (fluorescein) to deduce conservative solute transport paths. Digital images of fluorescein transport were captured under ultraviolet light and processed to analyse tracer plume geometry and obtain point-concentration breakthrough histories. Inorganic anion mixtures were also used to obtain conventional tracer breakthrough histories. Concentration data from the conservative tracer breakthrough curves was compared with the digital images and a well characterised numerical model. The results show that the peak tracer breakthrough response in dipole flow tracer tests samples a zone of aquifer close to the well screen, while the sampling volume of push-pull tests is limited by the length of the straddle packers used. The effective sampling volume of these single well forced-gradient tests in isotropic conditions can be estimated with simple equations. The experimental approach offers the opportunity to evaluate under controlled conditions the theoretical basis, design and performance of DFTTs and PPTs in porous media in relation to measured flow and transport properties. DFTT = Dipole Flow Tracer Test, PPT = Push Pull Tracer Test.

Barns, Gareth L.; Wilson, Ryan D.; Thornton, Steven F.

2012-02-01

401

Development of unique xanthene-cyanine fused near-infrared fluorescent fluorophores with superior chemical stability for biological fluorescence imaging.  

PubMed

The development of near-infrared (NIR) functional fluorescent dyes has gained increasing attention over the last few decades. Herein, we describe the development of a unique type of xanthene-cyanine fused NIR fluorophores, XC dyes, formed by reacting chloro-substituted cyanine with resorcin or its analogues under anhydrous conditions. XC dyes are a hybrid of cyanine and xanthene. The preliminary mechanistic studies indicate that the formation of XC compounds likely includes a sequence of cyclization and oxidation. XC dyes have absorption and emission in the NIR region, and their fluorescence properties can be controlled by modifications of the key hydroxyl and amine groups. The novel XC NIR dyes are advantageous over previously developed merocyanine dyes NIR dyes in their chemical stability against strong nucleophiles. Quantum chemical calculations reveal that the distinct properties of XC and HD dyes can be attributed to their structural differences. By taking advantage of the superior properties of XC dyes, we have further constructed a new NIR fluorescent probe, XC-H2 S, which is capable of monitoring both the concentration- and time-dependent variations of H2 S in living animals, highlighting the value of XC NIR dyes. We expect that the unique XC NIR dyes developed herein will find broader applications than HD NIR dyes as fluorescent platforms for the development of a wide variety of NIR fluorescent probes, in particular, those suitable for targets of interest that have strong nucleophilic character. PMID:25388080

Chen, Hua; Lin, Weiying; Cui, Haijun; Jiang, Wenqing

2015-01-01

402

Spore Stain of Bacillus cereus  

NSDL National Science Digital Library

This strain of Bacillus cereus was isolated from a sample of gasoline-contaminated soil and cultured on blood agar. This picture allows students to see spores utilizing a simple, reliable method of staining.

American Society For Microbiology;

2002-01-01

403

Normalization of Voltage-Sensitive Dye Signal with Functional Activity Measures  

PubMed Central

In general, signal amplitude in optical imaging is normalized using the well-established ?F/F method, where functional activity is divided by the total fluorescent light flux. This measure is used both directly, as a measure of population activity, and indirectly, to quantify spatial and spatiotemporal activity patterns. Despite its ubiquitous use, the stability and accuracy of this measure has not been validated for voltage-sensitive dye imaging of mammalian neocortex in vivo. In this report, we find that this normalization can introduce dynamic biases. In particular, the ?F/F is influenced by dye staining quality, and the ratio is also unstable over the course of experiments. As methods to record and analyze optical imaging signals become more precise, such biases can have an increasingly pernicious impact on the accuracy of findings, especially in the comparison of cytoarchitechtonic areas, in area-of-activation measurements, and in plasticity or developmental experiments. These dynamic biases of the ?F/F method may, to an extent, be mitigated by a novel method of normalization, ?F/?Fepileptiform. This normalization uses as a reference the measured activity of epileptiform spikes elicited by global disinhibition with bicuculline methiodide. Since this normalization is based on a functional measure, i.e. the signal amplitude of “hypersynchronized” bursts of activity in the cortical network, it is less influenced by staining of non-functional elements. We demonstrate that such a functional measure can better represent the amplitude of population mass action, and discuss alternative functional normalizations based on the amplitude of synchronized spontaneous sleep-like activity. These findings demonstrate that the traditional ?F/F normalization of voltage-sensitive dye signals can introduce pernicious inaccuracies in the quantification of neural population activity. They further suggest that normalization-independent metrics such as waveform propagation patterns, oscillations in single detectors, and phase relationships between detector pairs may better capture the biological information which is obtained by high-sensitivity imaging. PMID:19116673

Dann, Benjamin; Wanger, Tim; Ohl, Frank W.

2008-01-01

404

Coomassie blue staining for high sensitivity gel-based proteomics.  

PubMed

Gel electrophoresis, particularly one- (1DE) and two-dimensional electrophoresis (2DE), remain among the most widely used top-down methods for resolving and analysing proteomes. Detection of the resulting protein maps relies on staining (i.e. colloidal coomassie blue (CCB) or SYPRO Ruby (SR), in addition to many others). Fluorescent in-gel protein stains are generally preferred for higher sensitivity, reduced background, and wider dynamic range. Although traditionally used for densitometry, CBB has fluorescent properties. Indeed, infrared detection of CCB stained protein was comparable to SR, with BioSafe (Bio-Rad) and the Neuhoff formulation (NCCB) identified as potentially superior to SR; a minor sensitivity issue encountered in gel-resolved proteomes; might have been due to the unified staining protocol used. Here the staining protocol for both CCB formulations was optimised, yielding improved selectivity without affecting sensitivity; the resulting linear dynamic range was similar for BioSafe and NCCB and somewhat better than SR. 2D gel-based analyses of mouse brain and Arabidopsis thaliana (leaf) proteomes indicated markedly superior spot detection using the NCCB formulation. Thus more sensitive, quantitative in-gel protein analyses can be achieved using NCCB, at a fraction of the cost. PMID:23428344

Gauci, Victoria J; Padula, Matthew P; Coorssen, Jens R

2013-09-01

405

Fading of auramine-stained mycobacterial smears and implications for external quality assurance.  

PubMed

Light-emitting diode fluorescence microscopy is being scaled up for tuberculosis control, but fading of auramine-stained slides could compromise external quality assurance. We stored auramine-stained slides and reexamined them over time. Slides stored in all environments faded quickly, with significant changes in the proportion of positive slides in as little as 1 week. PMID:21430105

Minion, Jessica; Shenai, Shubhada; Vadwai, Viral; Tipnis, Tejashree; Greenaway, Christina; Menzies, Dick; Ramsay, Andrew; Rodrigues, Camilla; Pai, Madhukar

2011-05-01

406

Fluorescence Live Cell Imaging  

PubMed Central

Fluorescence microscopy of live cells has become an integral part of modern cell biology. Fluorescent protein tags, live cell dyes, and other methods to fluorescently label proteins of interest provide a range of tools to investigate virtually any cellular process under the microscope. The two main experimental challenges in collecting meaningful live cell microscopy data are to minimize photodamage while retaining a useful signal-to-noise ratio, and to provide a suitable environment for cells or tissues to replicate physiological cell dynamics. This chapter aims to give a general overview on microscope design choices critical for fluorescence live cell imaging that apply to most fluorescence microscopy modalities, and on environmental control with a focus on mammalian tissue culture cells. In addition, we provide guidance on how to design and evaluate fluorescent protein constructs by spinning disk confocal microscopy. PMID:24974023

Ettinger, Andreas

2014-01-01

407

Near Infrared Dyes as Lifetime Solvatochromic Probes for Micropolarity Measurements of Biological Systems  

E-print Network

Near Infrared Dyes as Lifetime Solvatochromic Probes for Micropolarity Measurements of Biological, metabolism, and excretion. With the recent widespread use of near-infrared (NIR) fluorescent dyes, ultraviolet/visible/near-infrared (NIR) absorption, and fluorescence (7,8) have been developed. The latter

Larson-Prior, Linda

408

Optimized excitation energy transfer in a three-dye luminescent solar concentrator  

Microsoft Academic Search

The spectral range of sunlight absorbed by a luminescent solar concentrator (LSC) is increased by using multiple dyes. Absorption, fluorescence, and fluorescence excitation spectra, and relative light output are reported for LSCs made with one, two, or three BODIPY dyes in a thin polymer layer on glass. Losses caused by multiple emission and reabsorption events are minimized by optimizing resonance

Sheldon T. Bailey; Gretchen E. Lokey; Melinda S. Hanes; John D. M. Shearer; Jason B. McLafferty; Gregg T. Beaumont; Timothy T. Baseler; Joshua M. Layhue; Dustin R. Broussard; Yu-Zhong Zhang; Bruce P. Wittmershaus

2007-01-01

409

Textile dye dermatitis.  

PubMed

The literature concerning textile dye dermatitis published during the last decade was reviewed. Sixty-one cases of dye-allergic contact dermatitis in which the presentation or course of the dermatitis was unusual or the dye allergen was one not previously reported have been described. The four new dye allergens discovered were Disperse Blue 106, Disperse Blue 85, Disperse Brown 1, and Basic Red 46. The incidence of dye dermatitis varied from 1% to 15.9% depending on the country, patient sample, and number of dyes in the patch test series. The 10 new dye allergens discovered in these studies were Disperse Blue 153, Disperse Orange 13, Basic Black 1, Basic Brown 1, the acid dyes Supramine Yellow and Supramine Red, the direct dye Diazol Orange, the basic dye Brilliant Green, Turquoise Reactive, and Neutrichrome Red. Disperse Blue 106 and Disperse Blue 124 were shown to be the strongest clothing dye sensitizers to date. Standard screening patch test series were found to be inadequate for the detection of textile dye sensitivity; therefore textile dye patch test series should be used. It is difficult to determine whether the incidence of dye dermatitis is increasing or decreasing because controlled epidemiologic studies are lacking, but data suggest that textile dye sensitivity is more common than previously believed. PMID:7896955

Hatch, K L; Maibach, H I

1995-04-01