Re: Tissue storage.

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From:"R.Wadley" <s9803537@pop3.unsw.edu.au>
To:HistoNet@Pathology.swmed.edu
Reply-To:
Date:Wed, 17 Mar 1999 18:26:15 +1000
Content-Type:text/plain; charset="us-ascii"

	Dear Ian,

	Have you considered museum techniques?

	Most Pathology Museum specimens are stored for long periods of time in low
formaldehyde concentration solutions.  In fact there is some evidence to
suggest that over time the formaldehyde can diffuse through the walls of
perspex pots, thus giving that charming(!?) concave effect to the large
surfaces.

	Wentworth's 5 solution for example is:
	Formaldehyde (40%)		20 ml
	Sodium acetate			40g
	Tri-sodium citrate			1g
	Glycerine				200 ml
	Distilled water			1000 ml
	
	Sodium hydrosulphite is also added (3 g/1000 g of tissue) to preserve colour.

	The glycerine of course is to make the solution a similar optical density
to perspex so can be ignored, the 2 sodium compounds = buffer set your pH
as you like it (7.5 for museum stuff).

	The other approach is to remove the formaldehyde from the tissue by
washing in running water, some authors suggest the addition of a little
soap to warm running water may help.  For un processed tissue section
washing may take only a couple of hours, 24 - 72 hours is recommended for
whole organ specimens.

	An alternative storage medium is a 1 - 10% detergent solution in distilled
water.  This system was/is used by the Queen Victoria Museum in Launceston
Tasmania.  I'm afraid I don't know much about it except the detergent is a
common household type, the company is Hunters, the product 'SAVLON'.

	Just a few ideas.

	Rob W.

At 09:24 AM 3/16/99 +0000, you wrote:
>	My group will be fixing small arteries at phyiological pressure
>then staining with nuclear dyes for confocal studies. After confocal
>microscopy some of the arteries will be processed for routine histology and
>ICC (endothelium, smooth muscle, elastin and collagen) while others will
>remain in a suitable fluid, just in case we need to repeat the confocal
>microscopy. In the past, the arteries were left in formalin as ICC was not
>required, but now it might be. Any suggestions as to a long term storage
>solution , bearing in mind that it might be at least 6 months or more
>before any decisions are made regarding further use.
>	I'm dealing with a bunch of Physiologists and Pharmacologists who
>will decide after 3-4 months that a particular specimen demonstrated a
>result which merits further study. I'll be led in, then asked to perform
>the necessary miracle be it histology, ICC or E.M. "Ah but, it was sitting
>in formalin for 6 months, it's nae use". "Why not, and why didn't you tell
>us earlier".

R. Wadley, B.App.Sc, M.L.S
Laboratory Manager
Cellular Analysis Facility
School of Microbiology & Immunology
UNSW, New South Wales, Australia, 2052
Ph (BH) 	+61 (2) 9385 3517
Ph (AH)	+61 (2) 9555 1239
Fax 	+61 (2) 9385 1591
E-mail	r.wadley@unsw.edu.au
www	http://www.unsw.edu.au/clients/microbiology/CAF.html
	(Under development)



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