Jonathan Sheehan wrote:

> Jim-
> Regarding your protocol:
> _____________________________________________
> >Tissue Clearing Procedure for thick slices
> >
> >  This procedure is used for clearing thick (300-500
>
>
------------------------------------------------------------------------
>
> µm) slices of rat brain
> >tissue which has been fixed with 4% paraformaldehyde  and is stored in
> >buffer.
> >1. Transfer the tissue to a glass container.
> >2. Wash twice with distilled water for 5-10 minutes.
> >3. Wash with graded alcohols (65%, 70%, 80%, 85%, 90%, 95% and absolute)

> >for 5-10 minutes each.
> >4. Wash with xylene for 5-10 minutes.
> >5. Wash with Histoclear® for 5-10 minutes.
> >6. Mount the tissue in a depression well slide with Methyl Salicylate
and
> >seal  the cover slip with clear nail polish.
> _____________________________________________
>
> Would this be suitable for use with an inverted scope? I have heard (only

> anecdotally) that methyl salicylate could solvate nail polish; this would
make
> me hesitate to work with it inverted over my objectives. Perhaps you can
dispel
> my misinformation. Or perhaps you use a specific brand of nail polish?
>
> Thanks,

 Jonathan ,

  This method is ok for inverted microscopes. Yes, the nail polish is
solubilized by the methyl salicylate. It happens slowly and the preps are
stable
for months. The trick is to blot the slide dry after cover slipping and
before
nail polishing to minimise the solvent interface. We've used nail polish
from EM
supply houses and also various brands from pharmacies, etc. and see no
difference
in performance. We have also mounted these tissue samples in Permount®
media with
similar results.


Don

--

Donald Szarowski                [log in to unmask]
Research Scientist              Office: 518-402-5233
Wadsworth Center  Fax:    518-473-2900
Box 509
Albany, NY 12201