Rapid wright giemsa staining solution

In this paper, we investigated the phagocytosis rate and phagocytosis index by Wright-Giemsa staining tests. Here, we chose primary murine peritoneal macrophages as effector cells, C. albicans SC5314 and 11g (0.0313 × 10^–3 mg/mL)-treated C. albicans SC5314 as target cells. Macrophages were cultured in 48-well plates at 37°C for 48 h. Overnight cultures of C. albicans SC5314 and 11g (0.0313 × 10^–3 mg/mL)-treated C. albicans SC5314 were harvested and washed three times with PBS. After discarding the culture medium of macrophages, C. albicans SC5314 or 11g (0.0313 × 10^–3 mg/mL)-treated C. albicans SC5314 was added to macrophages [multiplicity of infection (MOI) = 30] and cultured at 37°C for 0.5 h. After discarding the culture medium, 48-well plates were washed by PBS for two times. Cells were stained by rapid Wright-Giemsa staining solution (BBI Life Sciences Corporation, Shanghai, China). Then cells were washed by PBS for three times. Cells were photographed and counted by a microscope (Nicola and Casadevall, 2012 (link)). Phagocytosis rate = (number of macrophages phagocytizing fungi in 200 macrophages/200) × 100%; Phagocytosis index = number of fungi phagocytized by 200 macrophages/200.

Zebrafish embryos at 60 hpf were anesthetized by tricaine, then were immersed in mixture solution (40% FBS [HyClone] and a final concentration of 5 mM EDTA solution in 1X PBS). Red blood cells were collected from the heart using a microinjection needle. The cell solution was centrifuged at 1000 rpm for 5 min and the supernatant was discarded. Then the enriched cells were smeared evenly on slide and air-dried rapidly. After fixing in the methanol for 5 min, the slide was soaked in rapid Wright–Giemsa staining solution (BBI Life Sciences) for 10 min. Finally, slides were rinsed with deionized water. Images were captured with 100× lens after air dry. Assessment of erythrocyte status was based on previously published article^{34 (link)}.