RE: [Histonet] processing[Scanned]

From:Kemlo Rogerson

Wasn't there a procedure in Lillie's book about regenerating 'overcooked'
tissue? I fancy it involved using glycerol or something like that; I'll have
a scout around but I'm sure I used it when I was a pup.

-----Original Message-----
From: Gayle Callis [] 
Sent: 15 March 2005 20:53
To: Jennifer Sipes;
Subject: Re: [Histonet] processing[Scanned]


I doubt this would help but rather make it worse as you have already 
removed both the free water in tissue (what you want to do) but if you take 
it backwards through alcohols again, you are going to further remove more 
of the bound water (that you want to retain) on proteins.  The best thing 
to do is try soaking tissues after facing the block, with warm water then 
cool the block and cut.

Even if you rehydrated it and then reprocessed,  any water you put into 
tissue spaces is probably going to be removed again plus reexposure to heat 
of paraffin is going to compound the issue, cooked twice over - so to speak.

Best is rethink the processing schedule for your particular tissue and use 
shorter processing times in future.

At 01:22 PM 3/15/2005, you wrote:
>I have some tissue that I processed and it came out a little bit drier 
>than I thought it would.  Is it possible to take it backwards through the 
>processing and rehydrate it and then process it again, BUT at shorter 
>times to ensure that the tissue would be processed properly the second 
>time?  Thanks in advance!
>Happy St. Patrick's Day!
>Jennifer K. Sipes, RALAT
>Sr. Laboratory Technician
>Johns Hopkins University
>Ross 929
>720 Rutland Avenue
>Baltimore, MD  21205
>phone:  410-614-0131
>cell:     443-413-0853
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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