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CFSE Labeling to detect cell proliferation (CAT#: STEM-CBT-0008-WXH)

Introduction

The fluorescent dye hydroxyfluorescein diacetate succinimidyl ester(CFSE) is a membrane-permeable fluorescent dye that can label living cells.
When CFSE diffuses into the intracellular environment, endogenous Esterase can hydrolyze its acetic acid group. This form of CFSE molecule has high fluorescence activity and can produce green fluorescence when excited, but it no longer has membrane permeability. It can also interact with Free amine groups in intracellular cytoskeletal proteins react to form fluorescent protein adducts.
Therefore, when the cells divide and proliferate, the cytoplasmic proteins with fluorescence are evenly distributed to the second generation cells, so their fluorescence intensity will be reduced to half; The fluorescence intensity of the third-generation cells will be weaker than that of the second-generation cells. This phenomenon can be detected and analyzed by flow cytometry under the excitation light of 488nm, and the situation of cell division and proliferation can be further analyzed by detecting the continuous decrease of the fluorescence intensity of the cells.




Applications

• Detection of cell proliferation and cell division
• Estimation of the cell cycle
• Live cell detection
• Observing long-term cell activity

Procedure

1. Cell Treatment
2. Make cell suspension
3. CFSE incubation
4. Termination marker
5. Resuspend cells
6. Flow cytometry analysis/fluorescence microplate quantification/fluorescence microscope observation

Notes

Customer provided:
1. Cell type and name
2. Detection time node

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