Nine fluorescence parameter analysis on a four-color fluorescence activated flow cytometer.
BACKGROUND
In many cases a frequent monitoring of blood is important, e.g. during an infection. Often the availability of blood is the limiting factor.


METHODS
50 microl blood were stained and analyzed using a standard four-color cytometer. Percentages of leukocytes were calculated by FlowJo software.


RESULTS
Our protocol allows the differentiation of B cells, T cells, NK cells, neutrophils, and monocytes/macrophages in small volumes of blood.


CONCLUSIONS
Using nine fluorochrome-labeled antibodies and a specific gating strategy we were able to differentiate the main immune cells in minute amounts using one staining step.
German Research Centre for Biotechnology, Department of Experimental Immunology, Mascheroder Weg 1, D-38124 Braunschweig. urd@gbf.de
Ursula
U
Frischmann
Werner
W
Müller
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology; D001402:B-Lymphocytes / Q000737:chemistry / Q000166:cytology; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000166:cytology; D018414:CD8-Positive T-Lymphocytes / Q000737:chemistry / Q000166:cytology; D002452:Cell Count; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D007371:Interferon-gamma / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D007694:Killer Cells, Natural / Q000737:chemistry / Q000166:cytology; D007962:Leukocytes / Q000737:chemistry / Q000166:cytology; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000166:cytology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D009504:Neutrophils / Q000737:chemistry / Q000166:cytology; D012737:Sex Factors; D013194:Staining and Labeling / Q000379:methods
D000906:Antibodies; D000954:Antigens, Surface; D016753:Interleukin-10; D007371:Interferon-gamma