CFDA [5-(and 6)-Carboxyfluorescein diacetate] *Mixed isomers*
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 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 217.207 µL | 1.086 mL | 2.172 mL | 10.86 mL | 21.721 mL |
5 mM | 43.441 µL | 217.207 µL | 434.414 µL | 2.172 mL | 4.344 mL |
10 mM | 21.721 µL | 108.604 µL | 217.207 µL | 1.086 mL | 2.172 mL |
Molarity calculator
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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
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
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
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
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
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
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
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
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
Authors: Hu H, Dong Y, Feng P, Fechner J, Hamawy M, Knechtle SJ.
Journal: Transplantation (2003): 1075
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