Carboxyfluorescein succinimidyl ester (CFSE) is a fluorescent cell staining dye. CFSE is cell permeable and covalently couples, via its succinimidyl group, to intracellular molecules,[1] notably, to intracellular lysine residues and other amine sources. Due to this covalent coupling reaction fluorescent CFSE can be retained within cells for extremely long periods. Also, due to this stable linkage, once incorporated within cells the dye is not transferred to adjacent cells. CFSE is commonly confused with carboxyfluorescein diacetate succinimidyl ester (CFDA-SE), although they are not strictly the same molecule; CFDA-SE, due to its acetate groups, is highly cell permeable, while CFSE is much less so. As CFDA-SE, which is non-fluorescent, enters the cytoplasm of cells, intracellular esterases remove the acetate groups and convert the molecule to the fluorescent ester. CFSE was originally developed as a fluorescent dye that could be used to stably label lymphocytes and track their migration within ...

Large benthic foraminifera are unicellular calcifying reef organisms that can form symbiotic relationships with a range of different microalgae. However, the cellular functions, such as symbiosis and calcification, and other aspects of cellular physiology in large benthic foraminifera are not fully understood. Amphisorus kudakajimensis was used as a model to determine the detailed cellular characteristics of large benthic foraminifera. We used calcein acetoxymethyl ester (calcein AM) as a fluorescent indicator for live confocal imaging. We demonstrated that calcein AM is a useful fluorescent indicator to stain the fine network of reticulopodia and the cytoplasm in living A. kudakajimensis. We showed that at least two types of reticulopodia exist in A. kudakajimensis: the straight bundle of reticulopodia that spreads from the aperture and the fine reticulopodia along the surface of the aperture and chamber walls. The cytoplasm in outer chambers was highly branched and contained a few dinoflagellates. In

Anti-fluorescein antibodies are excellent model systems for studying the biochemical basis of molecular recognition because a prodigious amount of both physico-chemical and structural information is available for these antibodies. Furthermore, recombinant single-chain antibodies have been produced for several anti-fluorescein antibodies, and site-specific mutagenesis studies have defined the energetic contributions of a number of key active-site residues. In previous studies, we determined the three-dimensional structure of an antigen-binding fragment of a high-affinity anti-fluorescein antibody (4-4-20) in complex with fluorescein. These studies showed that fluorescein binds tightly in an aromatic slot and participates in a network of electrostatic interactions. In this report, we examine the role of electrostatic interactions in the 4-4-20 antigen-combining site by observing the effects of pH on the fluorescence of fluorescein and antigen-binding affinity. These studies showed that the salt ...

Animals. All mice were used at 6-8 weeks of age. C57BL/6 mice were purchased from the National Cancer Institute (Frederick, Maryland, USA). A breeding pair of lymphotoxin-α (LTα) knockout mice (on a mixed 129 × C57BL/6 background) were originally obtained from David Chaplin (Washington University, St. Louis, Missouri, USA) and have since been backcrossed for more than eight generations onto a C57BL/6 background. For some experiments, LTα knockout mice were splenectomized 2 weeks prior to immunization. Splenectomies were performed by staff of the Veterinarian Care Services at Yale University.. Antigens. L. major promastigotes of the WR309 substrain were maintained at 23°C in Schneiders Drosophila medium (GIBCO BRL; Invitrogen Corporation, Grand Island, New York, USA) supplemented with 20% FCS and 50 μg/ml gentamicin. For some experiments, L. major parasites were labeled with CFSE (5- and 6-carboxyfluorescein diacetate succinimidyl ester; Molecular Probes Inc., Eugene, Oregon, USA) using a ...

ANA M. MATA, ANN E. SCHOFIELD, JANE WOODBINE, ANTHONY G. LEE, J. MALCOLM EAST; Probing the nucleotide binding site of sarcoplasmic reticulum (Ca2+ -Mg2)-ATPase with anti-fluorescein antibodies. Biochem Soc Trans 1 December 1989; 17 (6): 1105-1106. doi: https://doi.org/10.1042/bst0171105. Download citation file:. ...

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The present investigation has demonstrated that ionizing radiation in the therapeutic dose range stimulates a transient cellular generation of ROS/RNS. Temporally coincident is a radiation-induced reversible depolarization of the mitochondrial ΔΨ and decrease in mitochondrial entrapped calcein fluorescence, both hallmarks of the mitochondrial permeability transition. The amount of ROS/RNS generated is relatively constant over the dose range tested, but the number of cells that respond increases with the dose. The radiation-induced ROS/RNS generation, ΔΨ depolarization, and calcein release are inhibited by CsA but not by the structural analogue, CsH. Overexpression of the Ca2+-binding protein calbindin 28K or treatment of cells with BAPTA/AM, an intracellular Ca2+ chelator, also effectively block radiation-induced ROS/RNS. The increased ROS/RNS generation observed with radiation is common to all of the cell types examined but for one important exception, the mitochondrial DNA-less ρo ...

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TRPV4 was originally identified as a plasma membrane channel activated by hypotonic cell swelling (Liedtke et al., 2000; Strotmann et al., 2000; Watanabe et al., 2002; Vriens et al., 2004; Loukin et al., 2010a). A decrease in extracellular osmolarity induces Ca2+ influx through TRPV4 channels (Güler et al., 2002; Liedtke and Friedman, 2003; Raoux et al., 2007; Phan et al., 2009). To determine whether hypotonicity modulates [Ca2+]RGCs, cells were exposed to saline solutions with osmolarity reduced from 280 to 190 mOsm. As illustrated in Figure 6A for an RGC loaded with the Ca2+-insensitive cell-volume indicator dye calcein AM, a reduction in osmolarity of the superfusing saline from 280 to 192 mOsm saline evoked sustained swelling of the cell. The resulting increase in cell volume was detected as a decrease in the intensity of calcein fluorescence (Fig. 6A, green trace). In contrast to hypotonic stimuli, no change in intracellular volume was observed during exposure to 100 μm glutamate.. Cell ...

Fluorescent Dyes , Fluorescent Physiological and Neurological Indicators , Calcein, AM *UltraPure Grade*, 5 mM solution in anhydrous DMSO; Calcein, AM is a cell-permeant and non-fluorescent compound that is widely used for determining cell viability. In live cells the non-fluorescent calcein, AM is hydrolyzed by intracellular esterases into the strongly green fluorescent anion calcein. The fluorescent calcein is well-retained in the cytoplasm in live cells. Hela cells stained with AnaSpec s Calcein AM fluorescent dye (Cat# AS-89201, AS-89202, AS-89203 and AS-89204).; MCF-7 cells grown in 8-well slide chamber for 2 days. Cells were washed twice before adding 1 uM Calcein AM (AnaSpec cat# AS-89202). Images acquired every 3 seconds and total elapsed time on video is 12 min.; C46H46N2O23

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Determination of β-glucan in carbohydrates was studied by detecting change of exciting wavelength and emission wavelength as sodium fluorescein was mixed with β-glucan, glucose, fructose and maltose. The results showed that if sodium fluorescein was mixed with β-glucan, UV-absorption of fluorescein sodium changed from 436.5 nm to 510.6 nm, fluorescence absorption also changed: exciting wavelength from 487.3 nm to 475.9 nm, emission wavelength from 517.6 nm to 510.6 nm, if sodium fluorescein was mixed with other carbohydrates such as glucose, fructose or maltose, no change happened for fluorescence absorption. The optimal proportion of sodium fluorescein and β-glucan was 6:1, the linear concentration range was 0.1 μg/mL～0.25 μg/mL, the regression equation: y = -2.014x + 1.075 7,R2 = 0.997 8, the purity of β-glucan in raw material was 95 % using such equation, comparing with 96 % by standard enzyme method.

TY - JOUR. T1 - Intracellular fluorescent staining with carboxyfluorescein. T2 - a rapid and reliable method for quantifying dye-coupling in mammalian central nervous system. AU - Rao, G.. AU - Barnes, C. A.. AU - McNaughton, B. L.. N1 - Funding Information: We thank E. Marder and R. Eaton for technical advice, J. Pickett-Heaps and R.J. Mclntosh for use of microscope facilities, and J. Baldwin for technical assistance. This research was supported by P.H.S. Grant AG03376.. PY - 1986/6. Y1 - 1986/6. N2 - Previous studies investigating electrotonic coupling in mammalian central nervous system have used the fluorescent marker Lucifer Yellow as an indicator of the presence of intercellular junctions between neurons. The fluorescent dye 5,6-carboxyfluorescein is known to have approximately 5 times the fluorescent yield of Lucifer Yellow. We have investigated the use of this dye as a potential alternative fluorescent marker on two types of neurons in the rat hippocampus in vitro. Unfixed hippocampal ...

China Food and Drug Administration (CFDA) has released a handful of draft documents about clinical trials of medical devices. The documents, which detail the process for on-site inspections and the products that are exempt from studies, continue the ongoing overhaul of the regulation of medical devices in China.. CFDA released the documents in separate posts that went out within days of each other. The first post covered three draft documents about inspections of medical device clinical trials. In one of the documents, CFDA provides a long list of points regulatory officials should check when inspecting the conduct of medical device clinical trials. Most of the points, such as whether the informed consent process meets the relevant requirements, are applicable to all clinical research programs. Others are more specific to the conduct of clinical trials of medical devices.. The regulator released the checklist alongside two other draft documents. One of the texts, referred to as annex 3, details ...

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Fluorescein with NDC 51662-1533 is a a human prescription drug product labeled by Hf Acquisition Co Llc, Dba Healthfirst. The generic name of Fluorescein is fluorescein.

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Many yeast samples from brewing contain extracellular debris and are small in size, making them difficult to count using a traditional hemocytometer or brightfield cell counter. This technical note will provide the information needed to accurately count yeast concentration and viability using the fluorescent dyes fluorescein diacetate (FDA) and propidium iodide (PI ...

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At the 2009 CFDA Awards, Tracey Ullman ribbed Marc Jacobs, and Jack White gave Anna Sui the Geoffrey Beene Lifetime Achievement Award.

Traditional fluorescein (FAM) fluorophore - universal dye useful for various applications, compatible with most of existing fluorescent instruments. All products are available as pure 5- or 6-isomers.

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FLUORESCEÍNĂ s. f. Colorant organic fluorescent, obținut din rezorcină și acid ftalic. [Pr.: flu-o-] - Din fr. fluorescéine.

In the present experiments on microdissected tubules of rabbit kidney we present a refined stop-flow method for determining the rate of HCO3- absorption (J(HCO3)) or H+ secretion (JH) that can be applied to isolated microperfused tubules. Using the pH-sensitive indicator dye BCECF (2,7-bis [2-carboxyethyl]-5[6]-carboxyfluorescein) the luminal perfusate pH is continuously ...

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Mesenchymal stem cells (MSCs) have been shown to alleviate acute lung injury (ALI) and induce the production of regulatory dendritic cells (DCregs), but the potential link between these two cell types remains unclear. The goal of this study was to investigate the effect and mechanism of MSC-induced regulatory dendritic cells in ALI mice. In vivo experiments, C57BL/6 wild-type male mice were sacrificed at different times after intratracheal injection of LPS to observe changes in lung DC maturation and pathological damage. MSCs, DCregs or/and carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled DCs were administered to the mice by tail vein, and flow cytometry was performed to measure the phenotype of lung DCs and T cells. Lung injury was estimated by the lung wet weight/body weight ratio and histopathological analysis. In vitro, Western blotting or flow cytometry was used to detect the expression of Notch ligand or receptor in MSCs or DCs after coculture or LPS stimulation. Finally, in vivo and

CD4|sup|+|/sup|CD25|sup|+|/sup|Foxp3|sup|+|/sup| Tregs control the immune response and maintain immune homeostasis. This study examined whether Tregs can affect mouse enteritis and the Foxp3 (Forkhead transcription factor) transcriptional pathway. Mouse CD4|sup|+|/sup|CD25|sup|+|/sup| Treg cells were labelled using CFSE (5,6-carboxyfluorescein diacetate succinimidyl ester) and transferred to enteritis model mice. The mice were randomly divided into an enteritis group, a Treg-infusion group, a Treg-inhibiting group, and a control group. Histopathology, ELISA, flow cytometry, western blot, immunohistochemistry, and immunofluorescence were performed. Our results demonstrated that CD4|sup|+|/sup|CD25|sup|+|/sup| Tregs were successfully transferred. The disease activity index (DAI) scores in the Tregs-infusion group were lower than those of the enteritis and Tregs-inhibiting groups. The number of goblet cells and inflammatory cells was reduced, and the levels of IL-1|i|β|/i|, TNF-|i|α|/i|, NO,

Objective: Metal alloys used in dentistry and in other biomedical fields may release nickel ions in the oral environment. The release of nickel might influence the normal biological and physiological processes, including tissue wound healing, cell growth and proliferation. The aim of this study was to evaluate in vitro the effects of nickel ions on cell cycle, viability and proliferation. Materials and Methods: Human osteosarcoma cells (U2OS) and human keratinocytes (HaCat) were exposed to different nickel chloride (NiCl2) concentrations (0 - 5mM) for various periods exposure. The viability of cultured cells was estimated by flow cytometry using Annexin V-FITC and Propidium Iodide (PI). Cell proliferation was evaluated by using carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometry. Finally, the effects of NiCl2 on cell cycle were assessed and quantified by flow cytometry. Statistical analysis was performed by means of ANOVA followed by Tukeys test. Results: NiCl2 induced ...

Eukaryotic cells have a protective plasma membrane, which restricts the free movement of molecules from the external environment to the internal environment. This study aims to computationally model the transport of fluorescein derivatives across the monolayer of human brain microvascular endothelial cells (HBMEC). The determination of plausible effective diffusion constants (Deff) will allow models to be built that could be useful beyond in vitro experimentation. Fluorescein-5-isothiocyanate (FITC) modeling produced a Deff range of 1E-20 to 5E-20 cm2/s at a 1 µm cell monolayer thickness and a Deff constant near 5E-29 cm2/s at a 5 µm cell monolayer thickness. Both fluorescein and sodium fluorescein (NaFl) modeling at the 1 and 5 µm thicknesses did not produce simulations that closely resembled the HBMEC in vitro model. Overall, it is possible that the fluorescent intensity noted with fluorescein and NaFl may be better explained by a mechanism other than passive diffusion. Simulations of FITC

0148] Likewise, at least for the microspheres used in incubation step (a), the signal precursor molecules may be low molecular weight substances selected from the group consisting of fluorophores and their derivatives, luminophores and their derivatives, chromophores and their derivatives, prosthetic groups, or redox active substances selected from redox mediators, electrode-active substances. Preferably, low molecular weight signal precursor molecules are fluorophores such as fluoresceins, cyanines, carbocyanines, rhodamines, xanthenes, diazo-dye based fluorescent substances, and small fluorescent aromatic and heteroaromatic molecules. Alternatively, the low molecular weight signal precursor molecules may be chromophores such as pyrazolone, anthraquinone, carotenoid and diazo- and monoazo, oxazine, indigoid, or riboflavine based dye substances. Most preferably, the low molecular weight signal precursor molecules are fluorescein derivatives such as fluorescein diacetate (FDA), fluorescein ...

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Colicin E1 induces the efflux of carboxyfluorescein and calcein from liposomes whose phospholipid composition is similar to that of Escherichia coli. This colicin action takes place at protein-to-liposome ratios and within pH ranges that are physiologically meaningful. Colicin-induced permeability of carboxyfluorescein is not limited to the initial phase of colicin membrane interaction but is sustained thereafter. Colicin E1 requires negatively charged phospholipids in the liposomal membrane in order to bind and induce efflux.

We have developed a quantitative assay to monitor the oxidative burst (H2O2 production) of polymorphonuclear leukocytes (PMNL) using single cell analysis by flow cytometry, and have examined whether PMNL respond to membrane stimulation with an all-or-none oxidative burst. During incubation with normal neutrophils, dichlorofluorescin diacetate diffused into the cells, was hydrolyzed to 2,7-dichlorofluorescin (DCFH) and was thereby trapped within the cells. The intracellular DCFH, a nonfluorescent fluorescein analogue, was oxidized to highly fluorescent 2,7-dichlorofluorescein (DCF) by PMNL stimulated by phorbol myristate acetate (PMA). That the oxidative product was DCF was shown by excitation/emission spectra and by mass spectrometry of the product from PMA-stimulated PMNL. Normal resting and PMA-stimulated PMNL oxidized 6.9 +/- 0.7 and 160 +/- 13 attomoles DCF per cell, respectively, in 15 min. Absence of calcium and magnesium ions and/or addition of 2 mM EDTA did not inhibit DCF formation ...

Sodium fluorescein leakage for mono-, co-, and tetraculture in transwell inserts with 0.4, 1 and 3 μm pore size. Medium containing 10 µg/mL of sodium fluores

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The first step in determining whether a fluorescent dye can be used for antibody labeling consists in collecting data on its physical interaction with the latter. In the present study, the interaction between the 2-(2-hydroxy-5-nitrobenzylidene)-1,3-indanedione (HNBID) dye and the IgG1 monoclonal mouse antibody anti-human heart fatty acid binding protein (anti-hFABP) has been investigated by fluorescence and circular dichroism spectroscopies and complementary structural results were obtained by molecular modeling. We have determined the parameters characterizing this interaction, namely the quenching and binding constants, classes of binding sites, and excited state lifetimes, and we have predicted the localization of HNBID within the Fc region of anti-hFABP. The key glycosidic and amino acid residues in anti-hFABP interacting with HNBID have also been identified. A similar systematic study was undertaken for the well-known fluorescein isothiocyanate fluorophore, for comparison purposes. Our results

Fluorescein dye disappearance. Easiest and most common, place fluorescein drops in each eye, Wait five minutes, Objectively grade remaining fluorescein. If it

The CFDA has entered into an alliance with the countrys tax department to set up a social credit system. CFDA is presenting the credit system as a way to build a more complete picture of the corporate integrity of the businesses it regulates, a resource it sees as helping focus its efforts on high-risk organizations.. By partnering with the Chinese state tax administration, CFDA is looking to gain access to resources and capabilities to set up and build out the database of drug company credit ratings it proposed late last year. CFDA will work with the tax department to set up a credit information sharing platform. The system will enable the two organizations to pool information on companies. As well as the financial information associated with credit scores in the West, the CFDA database will include details of the regulatory status of each company.. Full details of what CFDA plans to track are yet to emerge. In the government briefing document that outlined the social credit system, the ...

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Martin Hughes (Bioc) (mjgh at uk.ac.cam.bio.mbfs) wrote: : ,In article ,26lcir$n6j at bigboote.WPI.EDU, eeyore at wpi.WPI.EDU ( eeYORE ) writes: : ,,Subject: viability assays : , : [about wanting a viability assay for roots : ,,I have tried FDA (fluorescein diacetate) stain, but this is not quantitative : ,,or reliable for plant tissue. Anyone have any ideas or experience? : ,, Melissa : I would also like to add a request. I have been using FDA to assay : for viability in protoplasts, which is a normal recomended method. : I am trying (as a control) to kill off protoplasts. Unfortunately, : follwing a death treatment (30 min in 0.1% NaAzide, or 0.3% KCyanide)- : which I imagine should be pretty fatal to the cells, the FDA stain : still shows positive (this is also the case with even higher toxin : levels). Any suggestions for a more sensitive method of viability : staining would be appreciated. I used neutral red which is accumulated in the vacuoles of living cells with a characteristic ...

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Electroporation affects the transfer of fluorescein through inner membranes, but not the chorion. Embryos were incubated with 10 μg/ml fluorescein for 40 minut

Fluorescein is a fluorescent dye thats used in everything from eye exams to oceanography. Many researchers use fluorescein to...

Biomerica, Inc. (OTCBB: BMRA), a global provider of advanced diagnostic products for the early detection of medical conditions, today announced that it received two new China CFDA approvals for its Parathyroid Hormone (PTH) and Calcitonin products. The products will be distributed through a new China distribution channel unrelated to Biomericas current distribution channel in China

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Special note about Community Feature Content. Any content and/or opinions uploaded, expressed or submitted through any Community Feature or any other publicly available section of the Web Site (including password-protected areas), and all articles and responses to questions, other than the content explicitly authorized by the Company, are solely the opinions and responsibility of the person or entity submitting them and do not necessarily reflect the opinions of the Company. By way of example, any recommended or suggested use of products or services available from the Company that is posted through a Community Feature is not a sign of approval or recommendation by the Company. If you choose to follow any such recommendation you do so at your own risk.. Links to Third Party Sites. The Web Site may contain links to other websites on the internet. The Company is not responsible for the content, products, services or practices of any third party websites, including without limitation sites linked to ...

Subject: Re: fluorescein > >Storage should be in the dark, to prevent photo bleaching of FITC when >samples are collected, fixed, handled - etc. After cutting sections >must be in dark after mounting with antifade mounting media. Vector >has a nice one, Vectashield Hardset. > >She can probably do all three preparations, but remember that if she >fixes in formalin or paraformaldehyde, the tissue will have some >autofluorescence. If the FITC signal is stronger, brighter than then >dimmer autofluoresence, then all should go well. In general, the >microcapsules are huge! and very bright. > >I would think ingesting FITC might damage it (digestive enzymes, etc) >but not sure, there may be some pH and enzyme considerations. Have >her contact tech services at Molecular Probes, they usually have >answers for fluorochromes. > >We have used FITC labelled microcapsules injected into bovine tonsil, >removed tissue, snap freeze and cryosectioned and fixed sections >immediately with NBF. Some autofluorescence ...

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Fluorescein injection, USP is a sterile solution for use intravenously as a diagnostic aid. It is a dark reddish orange solution with a pH of 8.3 to 9.8 and an osmolarity of 572 to 858mOsm/kg.. ...

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