Stain for manual body fluid cell counts

[Jonna]

We currently use crystal violet in acetic acid to stain body fluid WBCs and lyse the RBS in a manual cell count. A new employee suggested we use New Methylene Blue as it stains the WBCs, but does not lyse the RBCs. Does anyone have experience with this?

Thanks

[SMILLER]

For cell counts on various fluids, we simply use a hemacytometer without any staining.  I would think that it is pretty common?

Scott

[Jonna]

Yes, most of the experienced techs do not use any stain, but we offer the stain until they get more confident looking at the cells on the hemacytometer.

[kimannez]

Some of our techs would use New Methylene Blue on manual spinal fluid counts to make it easier to differentiate the lymphs from the RBCs on the hemacytometer.  However, didn't CAP recommend this at one point for manual fluid counts that are being done on brightfield?

[Jonna]

I just looked up the CAP standard HEM.35395, thanks for the suggestion! It actually mentions methylene blue. We have always used either the acetic acid/crystal violet or slide correlation.

Red Cell Confirmation Techniques Phase I

There is an additional procedure beyond unstained bright-field microscopic visualization

of cells on the hemocytometer used when necessary to ensure the accurate distinction of

erythrocytes from other cell types.

NOTE: Suggested techniques include acid rinsing of the fluid sample to lyse erythrocytes after

initially counting all cells, the addition of a stain such as methylene blue to improve recognition of

non-erythrocytes, correlation with the number and proportion of cells on the cytospin preparation

or phase microscopy.