H&E LOOKALIKE stain for resin embedded bone

From:Gayle Callis

The toluidine blue/basic fuchsin stain was developed by Diane Sterchi, and
is published in J of Histotechnology, titled Microwaveable toluidine blue
stain for surface staining of undecalcified bone sections, 17(4):357-359,
1994. 

 It is an H and E "LOOK A LIKE" stain for undecalcified bone in PMMA
plastic.  Keep in mind, when you want something that is specifically
eosinophilic, basic fuchsin will probably not do nor mean the same thing as
for eosinophilic granules in eosinophils.  However the staining with the
modified T blue/BF is excellent for undecalcified bone.  Modified means the
stains are heated in a microwave, but slides are not MICROWAVED IN THE
STAINS- this can mess up PMMA plastic embedded sections.  This stain can
also be done on acid etched, thick ground undecalcified bone sections.  

Cracking with PMMA sections is due to softening of plastic (IF you do not
remove it) and will occur with xylene based mounting medias and xylene.  It
is best to not to use mounting media plus coverslip on top of PMMA
sections. For viewing and correction for refractive index on microscope,
just place a coverglass on top WITHOUT MOUNTING MEDIA, it should work very
well.  The same thing happens with glycol methacrylate plastic sections, or
don't run them through extensive dehydration, after eosin, rinse quickly
with 95% ethanol, air dry and mount a coverglass with mounting media, but
do not go through xylene or clearing agents.  



At 01:18 PM 10/24/01 -0400, you wrote:
>Hi to Rachel and histonet,
>                                         I work with resin embedded soft
>tissue with metal implants and I also had a brownish precipitate that
>made reading the slide for measurements next to impossible.  Also had
>cracking of the section sometimes no matter how fast I covered from
>water.  The solution to the precipitate issue I use is a modified stain
>I pick up at a workshop at NSH 2000. Workshop No. 13 entitled
>Preparation and Methods for Undecalcified Bone sections. The workshop
>was given by D. Brown and M.Hyten both of Eli Lilly & Co.  The stain is
>a modified Toluidine Blue/ Basic Fuchsin.  This stains like an H&E for
>me on soft tissue and should work for you on bone since the work shop
>was for bone in plastic !  
>I never was able to figure out what was causing the cracking !  Was it
>the alcoholic stains ?  Are there any good aqueous hematoxylins ?  Any
>help would be very much appreciated!
>thanks 
>evan
>edessasau@acri.com
>
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)





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