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.
Frischmann
Ursula
U
German Research Centre for Biotechnology, Department of Experimental Immunology, Mascheroder Weg 1, D-38124 Braunschweig. urd@gbf.de
Müller
Werner
W
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