Re: staining plastic-embedded sections

From:Robert Schoonhoven <>

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


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