I want to prepare NBT solution, but I don't know how much NBT powder to dissolve in PBS. Also, I don't have a BSA, can I do the NBT test without a BSA?
1. Take 100ul of blood and mix with 100ul of 0.2% nitro-blue tetrazolium in saline and 100ul of PBS. You may incubate this mixture at 37°C for 15 min and then leave it at room temperature for 15 min.
2. You may make smear on a glass slide and fix with methanol for 3 min and then stain with Pappenheim's stain for 3–5 min. Neutrophils that ingest the dye and have an oxidative burst create large black formazan deposits in the cytoplasm.
3. You may run this assay in conjunction with blood from healthy control subjects to identify any problems that you may face with specimen handling.