RE: Bone in plastic

you can not remove GMA

if you put your blocks in a dessicator for a while the water should be
removed and they may harden up

try using a really sharp tungsten carbide knife, i prefer D profile

the thinner you cut usually the fewer wrinkles you get, try cutting thinner

some people put ammonia in the water bath, or a little alcohol, i have never
found either to help

after picking the sections up off the water bath onto + slides place them
directly on a flat slide warmer/hot plate set at 65 dC for about 10 min.

if the wrinkling is occuring during staining or dehydration, do not use
ethyl alcohol, use isopropyl and get to 100% as quickly as possible, the
alcohol with water in it (95%) will cause the section to loosen or wrinkle
more than 100% or xylene

i mix my own gma because the kits are not consistant from batch to batch,
and during the summer they add more hydroqunione to keep it from
prepolymerizing which affects how it sets up, also if you mix your own you
can control how much plasticizer you put in it which determines how brittle
it is, from soft eraser like )too much, to very hard and brittle like, too
little, the initator can also be controlled if you make your own.  this is
pretty involved but really easy to make on your own, if you need a recipe
let me know.

patsy ruegg

-----Original Message-----
From: Su, Phy-Huynh
To: ''
Sent: 10/13/00 11:25 AM
Subject: RE: Bone in plastic

There is no implant in the bones, and the histologist before me bought
kit from Electron Microscopy Sciences.
I've tried to do the first batch, but it didn't work well.
I have problems with 
1.  Infiltration.
2.  Embedding and polymerization. Some blocks are softer than others at
cutting.  The microtome I use is sitting in an equipment room with
processor, embedding, and an autoclave machine in there.  I suspect that
steam pumping out from the autoclave increases too much the humidity in
room, thus soften the blocks, but I am not sure.  I'll look for room to
transfer out the microtome. 
3.  Cutting.  Oh boy, so many wrinkles.  They are horrible!!!  I used
Leica Polycut S, and steel knife.  Sections were put in a water bath
Temp), but till so many wrinkles!!!  What is wet cutting?  
4.  Do I have to store all the blocks in a dessiccant?
5.  Is there any way to get the tissues out of the plastic block (the
you can deparaffinize a block and re-embed again)?  I think for the poor
infiltrated ones, I need to re-infiltrate them.

Thank you a lot for any help.

> -----Original Message-----
> From: []
> Sent:	Wednesday, September 13, 2000 2:37 PM
> To:
> Cc:
> Subject:	RE: Bone in plastic
> Do you want to use Glycol Methacrylate or Methyl?  Do the sample have
> implants (metal types)?  If they have metal implants you will have to
> MMA.  I can help you with GMA.
> Patsy
> 		-----Original Message-----
> 		From:	Su, Phy-Huynh []
> 		Sent:	Tuesday, September 12, 2000 10:46 AM
> 		To:	'Histonet'
> 		Subject:	Bone in plastic
> 		Dear Histonetters:
> 		Experienced bone plastic histologists, please help!  My
> wants to switch
> 		from paraffin to plastic for bone and cartilage.  We
> have two block of tissues from an animal
> with
> 		Legg-Calf-Pelve like disease, and they were not fixed.
> dehydrated
> 		gradually in 40% EtOH at 4 degree C till 100%, and are
> in the
> 		refrigerator.  
> 		I have sent out our old tunsgten carbide knife for
> reconditioning. 
> 		Please advise on what chemicals I should use.  The
> are from femoral
> 		heads of 50 day old swine.  They are quite big.  My
> sections would
> 		normally occupy most of the microscopic glass slide.  It
> have cartilage,
> 		bone marrow, and some connective tissues on it.
> of plastic
> 		embedding, tricks, should do and shouldn't do, etc... in
> polymerizing,
> 		cutting to avoid wrinkles, and materials to coat the
> are greatly
> 		appreciated.  And deplasticizing techniques for IHC???  
> 		I can't damage these tissues, because they are very hard
> get.  Please
> 		help!
> 		And thank you a thousand times!  '
> 		Su

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