Mice were injected with thioglycollate and after 4 h peritoneal cells were harvested. Peritoneal cells from mice of same genotype were pooled for neutrophils purification by Percoll gradient. Neutrophils were aspirated out from the interphase of 63% and 72% of Percoll gradient.

2.5 × 10⁶ neutrophils were added to 1 × 10⁶ heat-killed candida and incubated for 15 min at 37°C. Trypan blue (1 mL) was added to each mixture and wet mount was prepared. At least 500 yeasts (phagocytosed and extracellular) were scored on the basis of trypan blue intake.

1.5 × 10⁶ neutrophils were preincubated with 25% mouse serum at 37°C for 5 min and then 1.0 × 10⁷ candida was added to the cells. The tubes were rotated for 60 min at 37°C. Thereafter methoxycholate was added to stop the reaction followed by methylene blue. At least 300 yeast cells in wet preps were counted under microscope distinguishing the viable (unstained) versus dead (stained) cells.

Data are presented as mean ± SD of 3 experiments.