Flowcytometric analysis of the phagocytosis of peripheral blood Leu-M3 positive cells

Abstract

An advanced flowcytometric technique for measuring phagocytosis of the Leu-M3 positive cells was developed. A 500 micro-litters of heparinized blood sample was mixed with 200 microlitters of fluorescent latex beads suspension and incubated at 37°C. The sample was then hemolysed and stained with FITC labelled monocyte-specific monoclonal anti-Leu-M3 antibody. Two color analysis was then applied, and the cells were discriminated by the positivity of Leu-M3 and the number of beads taken. The cells of each group were then sorted, and collected for morphological observations with non-specific esterase staining and for fluorescent microscopic observations. This method has the advantages of less blood required and easy processing of many samples at one time. The mean values of the phagocytosis of peripheral blood monocytes from health were 22.5±8.1% (1 hr), and 43.3±14.8% (2 hr).