CFSE | AAT Bioquest

CFSE

5-(and 6)-Carboxyfluorescein diacetate, succinimidyl ester; CAS 150347-59-4

CFSE (carboxyfluorescein diacetate succinimidyl ester) is a cell-permeable fluorescent dye that covalently labels intracellular proteins, enabling multi-generational cell proliferation tracking through progressive fluorescence dilution in flow cytometry.

Cell Proliferation Tracking: Halves in fluorescence with each cell division
Intracellular Protein Labeling: Cell-permeable dye covalently binds cytoplasmic proteins for long-term retention
488 nm Laser Excitation: Compatible with standard FITC channels in flow cytometry and microscopy
Alternative to CMFDA: Superior stability and lower toxicity compared to traditional proliferation dyes

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).

Catalog	Size	Price	Quantity
22022	25 mg	Price

Overview

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 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	179.385 µL	896.925 µL	1.794 mL	8.969 mL	17.939 mL
5 mM	35.877 µL	179.385 µL	358.77 µL	1.794 mL	3.588 mL
10 mM	17.939 µL	89.693 µL	179.385 µL	896.925 µL	1.794 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	557.46 g/mol	=	mL	x	mM	=	mmol

Citations

View all 10 citations: Citation Explorer

Comparative evaluation of bispecific antibody formats targeting PD-1 and LAG-3 for dual checkpoint blockade in cancer immunotherapy

Authors:

Wang, Jie and Shi, Ning and Zhao, Pinnan and Zhou, Yangyihua and Tian, Juan and Ma, Yaowei and Yang, Xuechen and Feng, Jiannan and Qiao, Chunxia and Li, Xinying and others,

Journal:

Biomedicine \& Pharmacotherapy (2025): 118583

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

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

Physical properties

Molecular weight	557.46
Solvent	DMSO

Spectral properties

Excitation (nm)	498
Emission (nm)	517

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200
CAS	150347-59-4

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Page updated on September 30, 2024