Home Immunology The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation
Immunology JoVE (Open Access) Citable · DOI

The Use of Carboxyfluorescein Diacetate Succinimidyl Ester (CFSE) to Monitor Lymphocyte Proliferation

DOI: 10.3791/2259-v
What you'll learn
  • Label mouse lymphocytes with CFSE fluorescent dye
  • Measure cell division by tracking fluorescence dilution
  • Apply CFSE labeling in vitro and in vivo
Protocol

Biopharma Insights CFSE covalently labels long-lived intracellular molecules with the fluorescent dye, carboxyfluorescein. As such, when a CFSE-labeled cell divides, its progeny have half the amount of fluorescence, which can thereby be used to assess cell division. This article describes the procedures typically used for labeling mouse lymphocytes with CFSE.

Difficulty
intermediate
Total time
~2–3 hours per sample batch (including incubation and wash steps)
Model organism
Mouse lymphocytes
Biosafety
BSL-1

Steps

1
Label lymphocytes with CFSE dye

Incubate isolated mouse lymphocytes with carboxyfluorescein diacetate succinimidyl ester (CFSE) to covalently label long-lived intracellular molecules with fluorescent carboxyfluorescein.

▶ 01:16
2
Apply CFSE method in vitro and in vivo

Employ the labeled lymphocytes to assess cell proliferation through fluorescence dilution, either in cell culture (in vitro) or within living animals (in vivo).

▶ 03:01
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