[Histonet] RE:Muscle Artifact

Wed Jul 29 14:17:28 CDT 2009

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Subject: Histonet Digest, Vol 68, Issue 38

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Today's Topics:

   1. Artifact in  Muscle bx's frozen for extended time (Sharon Allen)
   2. flash frozen tissue (Kalleberg, Kristopher)
   3. Mouse Eyes (Jo-Ann Bader, Ms.)
   4. SLIDE PRINTER (Janice Mitchell)
   5. Re: Mouse Eyes (Rene J Buesa)

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Message: 1
Date: Wed, 29 Jul 2009 09:27:24 -0500
From: "Sharon Allen" <SAllen <@t> exchange.hsc.mb.ca>
Subject: [Histonet] Artifact in  Muscle bx's frozen for extended time
To: <histonet <@t> pathology.swmed.edu>
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	<BB6ADCD4B7ABB045A09A7634AC15CC610BEC8077 <@t> hscxmsmx0010.ad.wrha.mb.ca>
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Hi,
I would like suggestions on how to eliminate or decrease the effects of extended storage of muscle bx's in a -70°C freezer.  We do approx. 150 muscle bx's /year in our Neuropathology Lab and we keep all frozen muscle bx's indefinitely, (we have them all from the 80's on). We are asked fairly regularly to go back to these cases for diagnostic & research purposes & find many of them are compromised, readable but not optimum,  I am assuming from drying out from months/years of freezing.
We freeze them using the isopentane/liquid nitrogen method on a pc of cork with OCT anchoring them.  We store them in a small plastic tube with a screw top.
I would like to know if anyone has further methods to eliminate this artefact.
Possibly sealing with OCT?  Placing water in the bottom of the tube?   Wrapping the muscle bx in Parafilm, tin foil etc?
What containers do you use to store them in if you do not get this artefact?
Suggestions for long term storage would be very much appreciated.
Thanks
Sharon Allen
Senior Neuropathology Technologist
HSC
WPG, MB, CA
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Message: 2
Date: Wed, 29 Jul 2009 10:31:39 -0400
From: "Kalleberg, Kristopher" <Kristopher.Kalleberg <@t> unilever.com>
Subject: [Histonet] flash frozen tissue
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Hello All,

I will be running a study upcoming and we have decided not to run FFPE as we normally do and have decided to use flash frozen.  My big concern is what is the proper way to fix all of these SKIN samples to use for IHC.  Any recommendations as to if prefix the samples in sucrose and then flash freeze in OCT is best or any recommendations as to if post fixing (in acetone??) the already sectioned, flash frozen sample in OCT is best.  Any and and all help will be greatly appreciated.  Thank you.

Kris Kalleberg    

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Message: 3
Date: Wed, 29 Jul 2009 11:38:05 -0400
From: "Jo-Ann Bader, Ms." <jo-ann.bader <@t> mcgill.ca>
Subject: [Histonet] Mouse Eyes
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
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	<76D119EF12C904418800ED67CCB2062905E2AB9071 <@t> EXMBXVS1.campus.mcgill.ca>
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We have a new project pending with mouse eyes.  The lens must remain intact.  What is the  best method to process, embed and cut them?  Paraffin or glycol methacrylate?

Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University
1600 Pine Ave W. 
Room 312
Montreal, QC, Canada
H3G 1Y6

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Message: 4
Date: Wed, 29 Jul 2009 11:42:27 -0400
From: "Janice Mitchell" <MITCHELLJA <@t> email.chop.edu>
Subject: [Histonet] SLIDE PRINTER
To: <histonet <@t> lists.utsouthwestern.edu>
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Anyone out there in Histoland have any experience with Thermo slide mate?  We are looking into purchasing a few.   Thanks for any input, Janice

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Message: 5
Date: Wed, 29 Jul 2009 09:26:48 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Mouse Eyes
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>,	" Ms.Jo-Ann Bader"
	<jo-ann.bader <@t> mcgill.ca>
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I used to cut entire eyes of fish and lizards embedded in paraffin with great results. The only thing is that on those (many years ago) days I used clearing with Canada balsam followed by a short wash in benzene (not very safe) before the paraffin infiltration.
René J.

--- On Wed, 7/29/09, Jo-Ann Bader, Ms. <jo-ann.bader <@t> mcgill.ca> wrote:

From: Jo-Ann Bader, Ms. <jo-ann.bader <@t> mcgill.ca>
Subject: [Histonet] Mouse Eyes
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Wednesday, July 29, 2009, 11:38 AM

We have a new project pending with mouse eyes.  The lens must remain intact.  What is the  best method to process, embed and cut them?  Paraffin or glycol methacrylate?

Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University
1600 Pine Ave W. 
Room 312
Montreal, QC, Canada
H3G 1Y6
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