Tracking of leukocyte recruitment into tissues of mice by in situ labeling of blood cells with the fluorescent dye CFDA SE

J Immunol Methods. 2004 Mar;286(1-2):69-78. doi: 10.1016/j.jim.2003.11.016.

Abstract

Models of inflammatory and immune diseases are extensively studied in mice with engineered genetic mutations, and tracking the recruitment of blood leukocytes into tissues is an important component of these studies. A direct in situ method for labeling the total pool of blood cells in mice by a single intravenous injection of the fluorescent dye CFDA SE (CFSE) is described. The fluorescence intensity of labeled cells initially declines, but remains stable after 4 h, enabling detection weeks after labeling. Labeled leukocytes can be tracked as they accumulate in lymphoid tissues and sites of inflammation and then be immunophenotyped for analysis by flow cytometry. This method is rapid, reproducible and simple to perform.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Movement / immunology*
  • Female
  • Flow Cytometry
  • Fluoresceins
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Immunophenotyping
  • Leukocytes / chemistry*
  • Leukocytes / cytology
  • Leukocytes / immunology*
  • Lymph Nodes / chemistry
  • Lymph Nodes / immunology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Specific Pathogen-Free Organisms
  • Spleen / chemistry
  • Spleen / immunology
  • Staining and Labeling / methods*
  • Succinimides / blood*

Substances

  • 5-(6)-carboxyfluorescein diacetate succinimidyl ester
  • Fluoresceins
  • Fluorescent Dyes
  • Succinimides