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CFSE

5-(and 6)-Carboxyfluorescein diacetate, succinimidyl ester; CAS 150347-59-4
CFSE is a cell permeable, green fluorogenic tracer useful for multi-generational proliferation assays in flow cytometry platforms, with an excitation peak at 498 nm and an emission peak at 517 nm.

Principle and mechanism

CFSE (carboxyfluorescein diacetate succinimidyl ester) is a non-fluorescent dye that can be used to track cell proliferation over multiple generations. It is cell permeable, able to pass through the membrane of live cells. Once inside, the acetate groups of the molecule are cleaved by non-specific intracellular esterases, resulting in a fluorescent product that accumulates in the cell. Non-viable cells do not have active esterases, nor do their leaky membranes retain the dye; therefore they remain non-fluorescent. The activated CFSE stain forms covalent bonds with free primary amine groups exposed on surface and intracellular proteins, such as lysine side chains. This means CFSE labeling can be used for tracing, counting and identification of specific cells across multiple divisions.

Excitation and emission

CFSE (fluorescein, CFDA, SE) is a fluorescent compound with an excitation peak at 498 nm and an emission peak at 517 nm. It is well excited by the 488 nm argon laser line. The emission can be monitored with a flow cytometer using the FL1 channel or FITC filter set.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of CFSE [5-(and 6)-Carboxyfluorescein diacetate, succinimidyl ester] *CAS 150347-59-4* 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 mM179.385 µL896.925 µL1.794 mL8.969 mL17.939 mL
5 mM35.877 µL179.385 µL358.77 µL1.794 mL3.588 mL
10 mM17.939 µL89.693 µL179.385 µL896.925 µL1.794 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

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Spectrum

Citations

View all 9 citations: Citation Explorer
Microalgal cell division tracking using CFSE
Authors: Pozzobon, Victor and Lagirarde, Jules and Arnoudts, Clarisse and Levasseur, Wendie
Journal: Algal Research (2024): 103501
Hypoxia-activated ADCC-enhanced humanized anti-CD147 antibody for liver cancer imaging and targeted therapy with improved selectivity
Authors: Qi, Fang-Zheng and Su, Hui-Shan and Wang, Bo and Qian, Luo-Meng and Wang, Yang and Wang, Chen-Hui and Hou, Ya-Xin and Chen, Ping and Zhang, Qing and Li, Dong-Mei and others,
Journal: MedComm (2024)
Restoration of miR-299-3p promotes macrophage phagocytosis and suppresses malignant phenotypes in breast cancer carcinogenesis via dual-targeting CD47 and ABCE1
Authors: Tong, Shoufang and Zhu, Yingli and Leng, Yeqing and Wu, Yunling and Xiao, Xingxing and Zhao, Wenfeng and Tan, Shuhua
Journal: International Immunopharmacology (2024): 111708
Interaction of Treponema pallidum, the syphilis spirochete, with human platelets
Authors: Church, Brigette and Wall, Erika and Webb, John R and Cameron, Caroline E
Journal: PloS one (2019): e0210902
Antigen presentation of the Oct4 and Sox2 peptides by CD154-activated B lymphocytes enhances the killing effect of cytotoxic T lymphocytes on tumor stem-like cells derived from cisplatin-resistant lung cancer cells
Authors: Zhang, Xueyan and Zhang, Yanwei and Xu, Jianlin and Wang, Huimin and Zheng, Xiaoxuan and Lou, Yuqing and Han, Baohui
Journal: Journal of Cancer (2018): 367

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
Page updated on September 30, 2024

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Physical properties

Molecular weight

557.46

Solvent

DMSO

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

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

CAS

150347-59-4
CFSE (fluorescein, CFDA, SE) is a fluorescent compound with an excitation peak at 498 nm and an emission peak at 517 nm. Other spectra of interest include: FITC (Fluorescein-5-isothiocyanate) and iFluor 488. CFSE (fluorescein, CFDA, SE) belongs to the following categories: Amine Reactive Dyes and Probes for Conjugation, Cell Proliferation Assays, Flow Cytometry Reagents, Fluoresceins, and Fluorescence Activated Cell Sorting (FACS).
CFSE (fluorescein, CFDA, SE) is a fluorescent compound with an excitation peak at 498 nm and an emission peak at 517 nm. Other spectra of interest include: FITC (Fluorescein-5-isothiocyanate) and iFluor 488. CFSE (fluorescein, CFDA, SE) belongs to the following categories: Amine Reactive Dyes and Probes for Conjugation, Cell Proliferation Assays, Flow Cytometry Reagents, Fluoresceins, and Fluorescence Activated Cell Sorting (FACS).
CFSE (fluorescein, CFDA, SE) is a fluorescent compound with an excitation peak at 498 nm and an emission peak at 517 nm. Other spectra of interest include: FITC (Fluorescein-5-isothiocyanate) and iFluor 488. CFSE (fluorescein, CFDA, SE) belongs to the following categories: Amine Reactive Dyes and Probes for Conjugation, Cell Proliferation Assays, Flow Cytometry Reagents, Fluoresceins, and Fluorescence Activated Cell Sorting (FACS).
Treponema pallidum-platelet interactions. (A) Flow cytometry isolates the co-localized platelet and treponeme populations by SSC and FSC gating. Dot plots of (B) CFSE-labeled heat-treated treponemes demonstrate reduced binding to human platelets labeled with PE/Cy5 anti-CD41a compared with (C) CSFE-labeled viable treponemes. (D) CFSE-labeled viable treponemes bound significantly more human platelets (mean = 55.13% &plusmn; 2.91 [SD] *P&lt;0.0001) compared with heat-treated treponemes (mean = 19.05% &plusmn; 2.29 [SD]) following co-incubation for 16 hours at 37&deg;C and ~ 5% oxygen. Results represent the mean of three independent experiments with statistical significance computed by two-way ANOVA, with a minimum of 3 replicates per sample type per experiment. (E) Darkfield microscopy FOV counts (20 random locations/slide) demonstrate viable treponemes bind significantly more human platelets (mean = 31.73% &plusmn; 1.19 [SD] *P&lt;0.0001) than heat-treated treponemes (mean = 6.47% &plusmn; 1.19 [SD]) following co-incubation at 5% oxygen at 34&deg;C for 48 hours. Results represent the mean of three independent experiments with statistical significance computed by unpaired two-tailed Student&rsquo;s t test, n = 3. (F) Heat-treated treponemes (top) are non-motile yet morphologically similar to viable treponemes (bottom). (G) Darkfield microscopy at 1000x magnification demonstrates platelet interactions are reversible. Image capture from video micrographs show edgewise attachment of a treponeme to an activated platelet (0.01 s to 8.10 s) which then detaches and moves away (9.05 s). Source:&nbsp;<strong>Interaction of Treponema pallidum, the syphilis spirochete, with human platelets </strong>by Church et al., <em>PLOS</em>, &nbsp;Jan. 2019.
Effects of GTM (1, 1/2 and 1/4 -fold serum concentrations) and probenecid (Prob, 80&thinsp;&mu;M) on the intracellular accumulation of 6-CF in CHO-hOAT1 cells (left) and 5-CF in HEK293-hOAT3 cells.(right). The effects of GTM on the uptake activity of hOAT1 and hOAT3 are shown in&nbsp;Fig. 6.&nbsp;GTM at 1/4&minus;, 1/2&minus; and 1-fold serum concentrations significantly reduced the intracellular accumulation of 6-CF, an OAT1 substrate, by 43.9, 41.9 and 58.1%, respectively, when compared with blank serum specimen at corresponding concentrations. Likewise, GTM at 1/2&minus; and 1-fold serum concentration significantly reduced the intracellular accumulation of 5-CF, an OAT3 substrate, by 36.4% and 31.3%, respectively, when compared to blank serum specimen at corresponding concentration. As positive control inhibitors of hOAT1 and hOAT3, probenecid (80&thinsp;&mu;M) significantly reduced the intracellular accumulation of 6-CF and 5-CF by 50.4 and 50.7%, respectively. These&nbsp;<em>in vitro</em>&nbsp;studies indicated that GTM significantly inhibited the uptake transport mediated by hOAT1 and hOAT3. *P&thinsp;&lt;&thinsp;0.05, **P&thinsp;&lt;&thinsp;0.01 and ***P&thinsp;&lt;&thinsp;0.001. Source: <strong>Green tea inhibited the elimination of nephro-cardiovascular toxins and deteriorated the renal function in rats with renal failure </strong>by Peng et al.,<em> Scientific Reports</em>, Nov. 2015.
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