Re: staining plastic-embedded sections

From:Philip Oshel <>

The url is:


>John is absolutely correct in that this is a lot for a beginner to take
>on.  Several of the stains mentioned will not work on JB-4, which is
>indeed glycol methacrylate (GMA).  Several years ago (OK, many years
>ago) I did a series of workshops at the NSH and Region IV meetings which
>included a rather lengthy handout on processing and staining tissues
>embedded in GMA.  I gave Steven Slapp at E-Beam Sciences permission to
>put it on their web site a few years ago and believe that most of those
>staining procedures are available there.  The I don't have the URL at
>hand (I'm not at my office computer), but a simple search should turn it
>up. If I can be of assistance please feel free to contact me.
>best regards,
>Robert Schoonhoven
>Laboratory of Molecular Carcinogenesis and Mutagenesis
>Dept. of Environmental Sciences and Engineering
>University of North Carolina
>Chapel Hill, NC 27599
>office 919-966-6343
>    Lab 919-966-6140
>    Fax 919-966-6123
>Don't go around saying the world owes you a living; the world owes you
>nothing; it was here first.
>Mark Twain [Samuel Langhornne Clemens] (1835-1910)
>"J. A. Kiernan" wrote:
>>  On Thu, 5 Oct 2000, Andrea Voogt wrote:
>>  > I'm a college student doing a research project on the histology of the
>>  > garter snake digestive tract and associated glands. I already have
>>  > tissues embedded in plastic, but all the books I can find have staining
>>  > procedures for paraffin.  Does anyone know the staining procedures for
>>  > plastic-embedded tissues?  The stains I'm hoping to use include
>>  > Mallory-Heidenhain's azan, Best's carmine, Gomori's chrome alum
>>  > hematoxylin-phloxine, Iron-hematoxylin and thiazine red staining,
>>  > Bielschowsky-Foot's method, and Masson's stain, but methods for any
>>  > stains that would be good for digestive tract and associated organs
>>  > would be much appreciated!!!
>>     This is a pretty tall order for a beginner! Some of those stains
>>     can be difficult to get right on paraffin sections (for which they
>>     are all intended), and you cannot simply apply the same procedure
>>     to plastic. You mention that the plastic is JB-4. This is glycol
>>     methacrylate (I think) and it has a chemically cross-linked molecular
>>     structure, so it cannot be dissolved out of the plastic. The
>>     embedding medium interferes with penetration of dyes and other
>>     reagents, and also with the dye-tissue specificities.
>>     Fortunately It is possible to cut plastic sections thinner than
>>     paraffin ones, and with a really thin section (say, 1 to 2 micrometres)
>>     you can see plenty of structural detail with just one dye that
>>     stains everything. An alkaline solution of toluidine blue is
>>     often used.
>>     There are published variants of older staining methods that you
>>     can use with glycol methacrylate embedding, and I'm sure you'll
>>     get plenty of good suggestions from the listserver, but I'd
>>     advise trying something very simple first, and not trying to
>>     bite off what might turn out to be too much to chew. Good luck.
>>   John A. Kiernan,
>>   Department of Anatomy & Cell Biology,
>>   The University of Western Ontario,
>>   LONDON,  Canada  N6A 5C1

Philip Oshel
Dept. of Animal Health and Biomedical Sciences
University of Wisconsin
1656 Linden Drive
Madison,  WI  53706-1581
voice: (608) 263-4162
fax: (608) 262-7420 (dept. fax)

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