[Histonet] Question regarding dehydration

[Julio Benavides]

Hi Ryan,

thanks a lot for your thoughts. These blocks were processed elsewhere 
and sent to us for the cutting and staining. Tissues were dehydrated in 
five consecutive baths of ethanol 100%, 5 hours each (manual 
processing). Then, they went to xilene (three baths, 4 hours each) and 
paraffin (two batch, 1h30´ each). Truth is that I have never seen such 
processing before. In our lab, with automatic processing, we begin with 
60% ethanol and go to 70, then 95 and then 100 before ethanol/xylene.

I was wondering if anybody has used the 100% ethanol processing before 
and which was the influence over tissues.

Thanks

Julio


-------- Forwarded Message --------
Subject: 	RE: [EXTERNAL] [Histonet] Question regarding dehydration
Date: 	Fri, 13 Feb 2015 09:06:47 -0500
From: 	Roy, Ryan <Ryan.Roy <@t> va.gov>
To: 	'Julio Benavides' <j.benavides <@t> eae.csic.es>



If the tissue is fatty it will blow apart on the water bath. If the H and E staining is poor its probably the processing.

What is the exact procedure for your process including time that the tissue was in each reagent? I would include this in a message to all of histonet as well as some may more knowledgeable than myself.

Also, I would recommend using a 95% solution prior to the 100% alcohol (s) if not a 70-80% and 95%.


Ryan Roy HTL (ASCP)
Manchester Veterans Affairs Medical Center
Manchester New Hampshire

Disclosure: The content of this email does not represent the views or opinons of the VA





-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Julio Benavides
Sent: Friday, February 13, 2015 4:26 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [EXTERNAL] [Histonet] Question regarding dehydration

Hi there,

we are having problems trying to cut some embedded samples (they crumble in the bath and the few cuts we manage to get into HE are crap). These are formalin fixed samples (bovine foetal and placenta samples) which went straight from formlin into 100º ethanol for the dehydration before clearing. I was wondering if such drastic dehydration step (no 60º, 70º or 90º ethanol before the 100º) could have damage the tissue. Has anyone have a similar issue before? do you think the samples are ruined for histology?

Thanks a lot for your help

Regards

Julio
Instituto de Ganaderia de Montaña
Spain

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