Although exactly the same procedure is done at the two different labs (in different cities) to permeate WBCs to cytoplasmic markers such as MPO or TdT, the preparation time differs significantly. The problem is that it usually takes more than 2 hours (after adding Triton) for one lab, but interestingly samples are ready in 10 mins in the other lab!
The only considerable difference between these two locations is their humidity!
That's the process I do:
- Add Paraformaldehyde 2% in PBS to the sample
- Keep at 2-4 C for 10 mins
- Add PBS and Centrifuge for 5 mins
- Add cold Triton to the sediment
- Put at 25 C till have a transparent solution
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