Re: JB-4 Plastic Embedding

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From:"R.Wadley" <> (by way of histonet)
To:histonet <>
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	Dear Grant,

	Your processing schedule will be work just fine if speed is
	Personally, I do not dehydrate at all, I take my specimen from
fixative to
JB4 infiltrating solution, using several changes until the tissue sample
sinks to the bottom of the infiltration container, then specimen can be
embedded.  Depending on the size & mass of the tissue infiltration can take
from 3 hours to 3 days.
	To polymerise I place the mould  over liquid nitrogen in a loosely
container, that is then refrigerated at 4C overnight.  To ensure complete
polymerisation around the top of the mould I follow up with an hour or so
under vacuum.  As the liquid nitrogen boils off it replaces the oxygen in
the container, thus providing anaerobic conditions.  Additionally it keeps
the curing resin cold, the polymerisation process is highly exothermic.  Do
not put uncured JB4 under vacuum as it boils rapidly and cures at 10-15
times its normal volume.
	While JB4 is clear it does colour with time to an orange/brown colour
which does not seem to interfere with staining or photography.
	I have not experienced any problems with swelling or shrinkage
except those caused by the fixative.
	As an alternative supplier I would suggest ProSciTech:

At 09:12 AM 1/27/99 -0800, you wrote:
>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 fininsh 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 soluable, clear and
>Polysciences says it can be done quickly.
>Any advice?
>Thanks Gordon

R. Wadley, B.App.Sc, M.L.S
Laboratory Manager
Cellular Analysis Facility
School of Microbiology & Immunology
UNSW 2052
Ph (BH) 	+61 (2) 9385 3517
Ph (AH)	+61 (2) 9555 1239
Fax 	+61 (2) 9385 1591
	(Under development)

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