CFSE

Description: CFSE is widely used for cell tracking and proliferation studies. It has also been used in CTL assays and cell motility studies. CFSE readily crosses intact cell membranes. Once inside the cells, intracellular esterases cleave the acetate groups to yield the fluorescent carboxyfluorescein molecule. The succinimidyl ester group reacts with primary amines, crosslinking the dye to intracellular proteins. Cell division can be measured as successive halving of the fluorescence intensity of CFSE. Cells labeled with CFSE may be fixed and permeabilized for analysis of intracellular targets using standard formaldehyde-containing fixatives and saponin-based permeabilization buffers, such as the Foxp3 Staining Buffer Set (cat. 00-5523) or the IC Fixation Buffer (cat. 00-8222) and Permeabilization Buffer (10X) (cat. 00-8333).

CFSE has a molecular weight of 557.47. After the acetate groups are cleaved, it has a peak excitation of 494 nm and peak emission of 521 nm. Each vial of CFSE may be reconstituted to a stock concentration of 10 mM with 90 uL of anhydrous DMSO; once reconstituted it should be protected from light and stored at -20°C; avoid freeze-thawing.

References: Parish CR, Glidden MH, Quah BJ, Warren HS. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr Protoc Immunol. 2009 Feb;Chapter 4:Unit4.9.

Ingulli E. Tracing tolerance and immunity in vivo by CFSE-labeling of administered cells. Methods Mol Biol. 2007;380:365-76.

Miller MJ, Wei SH, Parker I, Cahalan MD. Two-photon imaging of lymphocyte motility and antigen response in intact lymph node. Science. 2002 Jun 7;296(5574):1869-73.

Lyons AB, Parish CR. Determination of lymphocyte division by flow cytometry. J Immunol Methods. 1994 May 2;171(1):131-7.