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CFDA [5-(and 6)-Carboxyfluorescein diacetate] *Mixed isomers*

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Physical properties
Molecular weight460.39
SolventDMSO
Spectral properties
Excitation (nm)498
Emission (nm)517
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


See also: Fluoresceins
Molecular weight
460.39
Excitation (nm)
498
Emission (nm)
517
It is widely recognized that fluorescent labeling of cells is an effective means to determine total cell numbers or how many viable cells exist in a sample. Flow cytometry combined with fluorescent staining is a powerful tool to analyze heterogeneous cell populations. Fluorescein diacetate (FDA) and its derivatives are non-fluorescent molecules that diffuse into cells and are hydrolyzed by intracellular non-specific esterases to give fluorescent products. The fluorescent products can be accumulated only in those cells that have intact cell membranes; therefore, dead cells with leaky membranes are not stained. The precise kinetics of membrane transport and intracellular hydrolysis of FDA and its analogs (such as CDCFDA) are related to cellular functions, thus FDA labeling can be used for monitoring cells by flow cytometry or fluorescence microscopy. The fluorescence intensity of labeled cells by FDA dyes varies considerably among cell lines and strains, probably because of differences in intracellular esterase activity. CFDA is an amine-reactive FDA derivative that can be used to prepare a variety of FDA conjugates.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of CFDA [5-(and 6)-Carboxyfluorescein diacetate] *Mixed isomers* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM217.207 µL1.086 mL2.172 mL10.86 mL21.721 mL
5 mM43.441 µL217.207 µL434.414 µL2.172 mL4.344 mL
10 mM21.721 µL108.604 µL217.207 µL1.086 mL2.172 mL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Excitation (nm)498
Emission (nm)517

Images


Citations


View all 4 citations: Citation Explorer
Fluid and cell behaviors along a 3D printed alginate/gelatin/fibrin channel
Authors: Xu, Yufan and Wang, Xiaohong
Journal: Biotechnology and bioengineering (2015): 1683--1695
Overexpression of the CaTIP1-1 pepper gene in tobacco enhances resistance to osmotic stresses
Authors: Yin, Yan-Xu and Wang, Shu-Bin and Xiao, Huai-Juan and Zhang, Huai-Xia and Zhang, Zhen and Jing, Hua and Zhang, Ying-Li and Chen, Ru-Gang and Gong, Zhen-Hui
Journal: International journal of molecular sciences (2014): 20101--20116
Identification of VAR2CSA domain-specific inhibitory antibodies of the Plasmodium falciparum erythrocyte membrane protein 1 using a novel flow cytometry assay
Authors: Obiakor, Harold and Avril, Marion and MacDonald, Nicholas J and Srinivasan, Prakash and Reiter, Karine and Anderson, Charles and Holmes, Kevin L and Fried, Michal and Duffy, Patrick E and Smith, Joseph D and others, undefined
Journal: Clinical and Vaccine Immunology (2013): 433--442
Aging enhances maceration-induced ultrastructural alteration of the epidermis and impairment of skin barrier function
Authors: Minematsu, Takeo and Yamamoto, Yuko and Nagase, Takashi and Naito, Ayumi and Takehara, Kimie and Iizaka, Shinji and Komagata, Kazunori and Huang, Lijuan and Nakagami, Gojiro and Akase, Tomoko and others, undefined
Journal: Journal of dermatological science (2011): 160--168

References


View all 68 references: Citation Explorer
Novel method for cell debris removal in the flow cytometric cell cycle analysis using carboxy-fluorescein diacetate succinimidyl ester
Authors: Terho P, Lassila O.
Journal: Cytometry A (2006): 552
Modification of the fluorescein diacetate assay for screening of antifungal agents against Candida albicans: comparison with the NCCLS methods
Authors: Brouwer N, Kohen J, Jamie J, Vemulpad S.
Journal: J Microbiol Methods (2006): 234
Optimisation of the fluorescein diacetate antibacterial assay
Authors: Wan, undefined and y S, Brouwer N, Liu Q, Mahon A, Cork S, Karuso P, Vemulpad S, Jamie J.
Journal: J Microbiol Methods (2005): 21
A three-dimensional flow control concept for single-cell experiments on a microchip. 2. Fluorescein diacetate metabolism and calcium mobilization in a single yeast cell as stimulated by glucose and pH changes
Authors: Peng XY, Li PC.
Journal: Anal Chem (2004): 5282
Effect of immunosuppressants on T-cell subsets observed in vivo using carboxy-fluorescein diacetate succinimidyl ester labeling
Authors: Hu H, Dong Y, Feng P, Fechner J, Hamawy M, Knechtle SJ.
Journal: Transplantation (2003): 1075
Carboxy-fluorescein diacetate, succinimidyl ester labeled papillomavirus virus-like particles fluoresce after internalization and interact with heparan sulfate for binding and entry
Authors: Drobni P, Mistry N, McMillan N, Ev and er M., undefined
Journal: Virology (2003): 163
Detection of Saccharomyces cerevisiae carboxylesterase activity after native and sodium dodecyl sulfate electrophoresis by using fluorescein diacetate as substrate
Authors: Lomolino G, Lante A, Crapisi A, Spettoli P, Curioni A.
Journal: Electrophoresis (2001): 1021
Fluorescein diacetate hydrolysis as a measure of fungal biomass in soil
Authors: Gaspar ML, Cabello MN, Pollero R, Aon MA.
Journal: Curr Microbiol (2001): 339
Extrusion of fluorescein diacetate by multidrug-resistant Candida albicans
Authors: Yang HC, Mikami Y, Imai T, Taguchi H, Nishimura K, Miyaji M, Branchini ML.
Journal: Mycoses (2001): 368
Potential problems with fluorescein diacetate assays of cell viability when testing natural products for antimicrobial activity
Authors: Clarke JM, Gillings MR, Altavilla N, Beattie AJ.
Journal: J Microbiol Methods (2001): 261