Re: JB-4 Plastic Embedding

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From:"Bill Sinai (Anatomical Pathology)" <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

Date:          Wed, 27 Jan 1999 09:12:31 -0800
From:          "Grant, Gordon" <>
Subject:       JB-4 Plastic Embedding
To:            "''" <>

When using plastic embedding/resin embedding for large specimens I
found dehydration in graded acetone much more effective than ethanol
which is not always 100% (usually a little water).
A typical protocol would be: depending on tissue size:
70% acetone  1-2hours
80% acetone  1-2hours
95% acetone  1-2hours
100% acetone 2 changes of 1-2 hours.
uncatalysed plastic or resin to impregnate under vacuum
1-2 hours
then embed.

I think you will find many techniques in Electron Microscopic texts
or protocols.
 Good evening
Histonetters I have been asked to deploy a metallic stent in a
coronary artery then process for plastic embedding. The specimen will
then be cured, sawed in half and polished to a clear finnish and
evaluated under a dissection microscope. I found in Poysciences a
JB-4 embedding kit. My question is to you all is what dehydration and
infiltration technique should I use? I was thinking of using the
processing schedule below: 1	50% ETOH		30 minutes 2	60%
ETOH		30
minutes 3	70% ETOH		30 minutes
4	85% ETOH		30 minutes
5	95% ETOH 		30 minutes
6	100% ETOH		30 minutes
7	100% ETOH		30 minutes
8	100% ETOH		30 minutes
9	100% ETOH/JB-4	4 hours
10	JB-4			overnight
11	Embed
It looked intriguing because it is water soluble, clear and
Polysciences says it can be done quickly.
Any advice?
Thanks Gordon

Bill Sinai
Department Manager
Tissue Pathology
ICPMR Westmead Hospital
Phone 61+2+9845 7774  Fax 61+2+9687 2330

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