Tracking of leukocyte recruitment into tissues of mice by in situ labeling of blood cells with the fluorescent dye CFDA SE.
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.
Becker
Henry M
HM
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto General Research Institute, Toronto General Hospital, 200 Elizabeth Street, Eaton-4, Toronto, Ontario, Canada M5G 2C4.
Chen
Mian
M
Hay
John B
JB
Cybulsky
Myron I
MI
D000818:Animals; D002465:Cell Movement / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D005452:Fluoresceins; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D016130:Immunophenotyping; D007962:Leukocytes / Q000737:chemistry* / Q000166:cytology / Q000276:immunology*; D008198:Lymph Nodes / Q000737:chemistry / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013047:Specific Pathogen-Free Organisms; D013154:Spleen / Q000737:chemistry / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods*; D013388:Succinimides / Q000097:blood*
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides