I've been testing a protocol to evaluate stained whole blood in a wet mount for a WBC differential using Toluidine Blue, which results in clumping.
The protocol requires a 1:50 ratio of whole blood mixed with normal saline, then a 1:1 ration of blood dilution to stain incubated at 37C for 5 minutes. 10ul are placed into a disposable hemocytometer for inspection.
The staining is perfect however the cells clump together creating difficulties in count and differentiation.
A blood smear is not an option in this particular scenario. Alternative stains for a WBC differential using a wet mount could be an option.
Any guidance would be appreciated. Thank you.
Dear D. Kresser,
Clumping of whole blood in a wet mount stained with Toluidine Blue can be reduced by taking certain precautions and optimizing your staining technique. Here are some steps you can follow:
Thank you
The question was for a wet mount preparation, not a blood smear. It would also not use a fixative.
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