GRANULOCYTE FUNCTION TEST STIMULATED BY STAPHYLOCOCCUS AUREUS, FDA 209P JC-1 AND ESCHERICHIA COLI, NIHJ JC-2, USING A FLOWCYTOMETER

1. STUDY OF TESTING CONDITIONS

Abstract

The purpose of this research was to evaluate granulocyte function rapidly with small amount of heparinized blood under the condition close to biological response. The asaay of hydrogen peroxide (H₂O₂) generation of granulocyte stimulated by S. aureus FDA 209P JC-1, and E. coli, NIHJ J C-2, using a flowcytometer has been developed.
Magnitude of H₂O₂ generation of granulocyte which was incubated for 24 hours at room temperature, was similar to that of those treated just after bleeding.
Optimum concentration of bacterial solution was OD 0.6 at 620nm with a spectrophotometer. The asaay was performed as follows; after 60 minutes, incubation of the mixture at 37°C containing of 50μl of heparinized whole blood and 50μl of bacterial solution, the mixture was put into cold water at 4°C to suspend the reaction and measured H₂O₂ using a flowcytometer within six hours after suspension.