Use of the Intracellular Fluorescent Dye CFSE to Monitor Lymphocyte Migration and Proliferation

Abstract

The stable incorporation of the intracellular fluorescent dye 5-(and -6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) into cells provides a powerful tool to monitor cell migration, and to quantify cell division, because of the sequential decrease in fluorescent labeling in daughter cells. CFSE-labeled lymphocytes have been used to analyze the relationship between cell division and differentiation of cell function, and cell proliferation versus apoptosis, both in vivo and in vitro, and have allowed analysis of the site of response to antigens in vivo.

Fluorescent target array killing assay: A multiplex cytotoxic T‐cell assay to measure detailed T‐cell antigen specificity and avidity in vivo

“…We have previously observed a toxic effect when cells are labeled with CFSE, however, this can be buffered against by incorporating protein into the labeling solution (15,31). Indeed, more recently we have shown that CFSE, CTV, and CPD can be used at a combined concentration of up to 120 lM to label lymphocytes, without any effect on viability or function, provided labeling is undertaken in a buffer containing a high protein content (15).…”

Assessment of Protection Induced by DNA and Live Vaccine Encoding Leishmania MHC Class I Restricted Epitopes against L. major Challenge in Balb/c Mice Model

“…Cells were properly washed twice to remove un-integrated CFSE and restimulated with relevant peptides (10 µg/ml), irrelevant peptide (10 µg/ml) and ConA (5 µg/ml). Cells without CFSE staining were used as control [26]. 48 h later cells were re-harvested and stained for CD3 (PE-Hamster anti-mouse CD3e, BD biosciences) and CD8 markers (perCP Rat anti-mouse CD8α, BD biosciences) after 30 min incubation at 4°C in staining buffer.…”

Three Epstein–Barr virus latency proteins independently promote genomic instability by inducing DNA damage, inhibiting DNA repair and inactivating cell cycle checkpoints

“…Cell cycle and cell division Cell division was assessed by vital labeling using carboxyfluorescein diacetate succinimidyl ester (Molecular Probes, Invitrogen) (Parish et al, 2009) and DNA content was examined using propidium iodide staining.…”