Re: Internal Elastic Lamina and PMMA in general

<< Previous Message | Next Message >>
From:Gayle Callis <>
To:Taffy <>,
Content-Type:text/plain; charset=us-ascii

I do not know of any elastic stain that will work on thick PMMA sections,
unless you can modify from paraffin to plastic and etch the plastic to make
stain penetration deeper. PMMA just does not permit large molecule dye
penetration, and there only a few dyes that will do the job well, toluidine
blue, MacNeals tetrachrome made inhouse, Stevenels blue which is a
potassium permanganate oxidized methylene blue to produce t blue, thionin,
azures and methylene blue/basic fuchsin method.   Basic fuchsin, light
green, acid fuchsin and Van Giesons all work.  

You need to get into Richard Reckers book on Bone Histomorphometry and
Colin Anderson's book Manual on the Examination of Bone, and Method of
Calcified Tissue Preparation by GR Dickson and check out staining methods
to see if they will show the fibers you need to see.  Dickson is out of
print, but could be obtained via interlibrary loan in some way. Digging
through the journals as suggested by Cathy Mayton and Linda Jenkins for
PMMA on bone (Sorry Linda, but I have not published on PMMA, but Diane
Sterchi and Cathy Sanderson (Mayton) have numerable papers) is the other
alternative, been there, done that, learned a lot, and used a bunch of

J of Histotechnology, 20th anniversary issue, Sept 1997 has an article by
Cathy Sanderson (Mayton) which is an excellent review of PMMA on bone,
HOWEVER these staining technics will demonstrate SOFT TISSUES and it may
come down to you having to reinvent the wheel to make them work for what
you want.  So goes PMMA work, not easy at best.

Kulzer had a series of stains put out on for their Exakt system brochure
some years ago, and these may also work. 

Unles you can keep the ground 70 um section on a slide AFTER total removal
of PMMA with warm xylene or some other solvent that does the job, you may
not be able to use some of the tried and true PMMA staining methods.  This
is a tough call, and I wish you good luck 

At 02:12 PM 8/28/00 -0500, you wrote:
>Hello AGAIN Histonetters,
>I have a big problem..PLEASE help, if you can..
>I have an investigator that is looking for a stain for the internal Elastic
>Lamina (IEL) in THICK (70um) plastic sections (yep, this is still the
>artery)..they are wanting something that will stain this so they can do
>histomorphometry..ANY ideas?????????
>Thanks again for your help!!! 
>Taffy Hooser
>Mary (Taffy) Hooser, HT(ASCP)
>Department of Anatomy and Cell Biology
>Indiana University School of Medicine
>MS-5035, 635 Barnhill Drive
>Indianapolis, IN. 46202-5120
>Tel: (317)274-2308.
>Fax: (317)278-2040.
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

<< Previous Message | Next Message >>