[Histonet] dehydration after staining

[Gudrun Lang]

Hi all,

Is it possible to over-dehydrate the slides after staining?

 

We do our regular HE-stain with increasing ethanol-concentrations for 1-2
min and then clear in butyl-acetat before coverslipping with Pertex and
glass-coverslips.

One of our pathologists complains about an glassy-3D-appearence of the
tissue, especially of collagenfibers and mamma. The effect is most visible
in his microscope, which is the newest (2002), but only with the 4er and
10er objective. When he switches the condensor in, the effect is gone, but
there is only the small bright space. He compared it to older slides
(cleared in Xylol or Limonen, coverslipped with film or glass) and saw the
same effect in a milder way. The other pathologists don't see the same with
their (older) microskopes.

Could this be a matter of the microscope or a matter of clearing? Or a
matter of wrong handling of the microscope?

 

I would be happy for any input.

 

Gudrun Lang