[Histonet] dehydration after staining

[Geoff McAuliffe]

    Sounds like a poorly aligned (or dirty) microscope to me. Who looks 
at slides with the conderser out? (answer: people who don't understand 
microscope optics). I don't see how one can "over-dehydrate" a slide. If 
you leave some water in it will form bubbles as it separates from the 
mounting media.
    Good luck!

Geoff

Gudrun Lang wrote:

>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
>
> 
>
> 
>
>
>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff <@t> umdnj.edu
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