5-Carboxyfluorescein diacetate N-succinimidyl ester is a cell permeable dye (Ex=492 nm, Em=517 nm). 5-Carboxyfluorescein diacetate N-succinimidyl ester can label cells by covalently binding to intracellular molecules. 5-Carboxyfluorescein diacetate N-succinimidyl ester is used to track lymphocyte migration and proliferation[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].1. Dilute the stock solution in PBS (to give a 50 μM solution).2. Add 110 μL of this solution per milliliter of cells (to give a final concentration of 5 μM) and mix rapidly.3. After 5 min at room temperature, add 10 vol of PBS containing 5% FBS. 4. Centrifuge cells 5 min at 300 ×g, 20 ℃.5. Remove the supernatant, and wash threetimes, each time by resuspending in 10 vol PBS containing 5% FBS. 6. Centrifuge cells 5 min at 300 × g, 20 ℃, and removing the supernatant. Note: Labeling occurs rapidly, and it is essential that the tracker is dispersed as evenly and quickly as possible so that cells are uniformly labeled.
[1]. Christopher R Parish, et al. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr Protoc Immunol. 2009 Feb;Chapter 4:Unit4.9.