Re: [Histonet] Problem with Vectashield Hard Set

From:Gayle Callis


What you need to do with Vectashield Hard Set and also with Molecular 
Probes Prolong Gold  antifade, ready to use (hard set) is seal the edges of 
the coverslip with nail polish (only if you do not have GFP in the tissue 
section).  These hard set mounting medias  retract, as you describe, when 
they dry.  Sometimes the inserts do not tell you these little things -  I 
learned of this from both companies tech services.

We like to dump the clear nail polish out, rinse the nail polish bottle 
with acetone, let dry then put toluene or xylene diluted mounting media in 
the little bottle  since we like the tidy little brush for application to 
coverglass edges.   When we have GFP or DsRed present in tissue section, we 
use the diluted mounting media - since it does NOT contain alcohol.

At 09:42 AM 11/30/2005, you wrote:
>A couple of weeks ago I inquired about aqueous coverslipping media that
>harden. Vectashield Hard Set was recommended by a few people. Then I
>remembered that I actually had some of the product, which I purchased a
>couple of months ago, in my refrigerator.  I tried it, and at first was very
>pleased with it.  It seemed to have a refractive index closer to that of
>glass than most aqueous products, and the resolution was excellent.
>However, I am now finding that after being coverslipped for a couple of
>weeks a large amount of air space develops, first immediately around the
>tissue section, and eventually extending onto the tissue section.  This does
>not appear to be the typical problem of air being drawn under the edges of
>the coverslip due to evaporation of solvent, as there is no air space near
>the edges, only on and immediately around the tissue.  Besides, the stuff is
>supposed to harden overnight, according to the package insert.
>Has anyone else encountered this problem?  What is the nature of the
>problem?  Are the spaces actually air?  Or some evaporated component of the
>product?  And any hints on how to deal with it, other than soaking off the
>coverslips and recoverslipping?
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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