CFDA-SE is a fluorescent dye that can penetrate the cell membrane. It can react with the free amine group in the cytoskeleton protein inside the cell, and finally form a protein complex with fluorescence. After entering the cell, CFDA-SE locates in the cell membrane, cytoplasm and nucleus, and the fluorescence staining is strongest in the nucleus^[1]. CFDA-SE dye can be uniformly inherited by the cells with cell division and proliferation, and its attenuation is proportional to the number of cell divisions. This phenomenon can be detected and analyzed by flow cytometry under the excitation light of 488 nm, and can be used to detect the proliferation of cells^[1].

Guide (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
Preparation of CFDA-SE working solution
1.1 Preparation of the stock solution
Dissolve 1 mg of CFDA-SE in 0.1794 mL of DMSO to obtain 10 mM of CFSE.
Note: It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of CFDA-SE working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 5-10 μM of CFDA-SE working solution.
Note: Please adjust the concentration of CFDA-SE working solution according to the actual situation.
Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of CFDA-SE working solution, and then incubate at room temperature for 30 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Banks HT, et al. Quantifying CFSE Label Decay in Flow Cytometry Data. Appl Math Lett. 2013 May 1;26(5):571-577.  [Content Brief]

  [2]. A Bruce Lyons, et al. Flow cytometric analysis of cell division by dilution of CFSE and related dyes. Curr Protoc Cytom. 2013;Chapter 9:Unit9.11.  [Content Brief]

  [3]. Cui J, et al. Prostaglandin E3 attenuates macrophage-associated inflammation and prostate tumour growth by modulating polarization. J Cell Mol Med. 2021 Jun;25(12):5586-5601.  [Content Brief]

  [4]. Xiuyun Jiang et al. Long-lived pancreatic ductal adenocarcinoma slice cultures enable precise study of the immune microenvironment. Oncoimmunology. 2017; 6(7): e1333210.  [Content Brief]