Nine fluorescence parameter analysis on a four-color fluorescence activated flow cytometer

Abstract

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 μl 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. © 2006 International Society for Analytical Cytology.