RE: [Histonet] Re: Liquid nitrogen, mouse spleen - Freezing tissues

From:"Alan Bright"

Dear Andrea/ Gayle,

We have been manufacturing metal moulds for our cryostats and have found
the same, the front face of the block had been perfect for the Mohs
technique too.

Best Regards

Alan Bright

Bright Instrument Co.Ltd.
St Margaret's Way
Huntingdon
Cambridgeshire
PE29 6EU
England

Tel No:+44 (0)1480 454528
Fax No:+44 (0)1480 456031
Email: abright@brightinstruments.com
Web Site: www.brightinstruments.com
Skype User ID: dazzle0


-----Original Message-----
From: Gayle Callis [mailto:gcallis@montana.edu] 
Sent: 04 January 2007 18:27
To: Andrea Hooper; Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: Liquid nitrogen, mouse spleen - Freezing tissues


Dear Andrea and all,

Hooray for liberation!   We will try the metal sterilization trays 
and  what about disposable metal weighing dishes?   I think these are
out 
in  vendor land too.  Will be havet to go shopping.   Another key issue 
with  isopentane also know as 2 methyl butane is the storage problem.
The 
"floating boat" method eliminates a rather nasty solvent, although you 
still need to use Liquid nitrogen with good ventilation.

Gayle Callis

At 11:01 AM 1/4/2007, you wrote:
>I second Gayle's technique described below. It is wonderful. After
>spending so many years fixated on isopentane in liquid nitrogen and 
>getting cracked blocks in order to adhere with what was histologically 
>correct, I have finally moved onto the floating boat technique and my 
>frozens look nearly as good as paraffin sections. The only difference
is 
>that I use metal floating boats (read: sterilization trays) instead of 
>plastic petri dishes. This way the blocks cool even quicker - better
for 
>the tissue and morphology - though they still don't shatter.  I finally

>feel liberated :)
>
>
>
>At 9:27 AM -0700 1/4/07, Gayle Callis wrote:
>>Yes, your very last suggestion will work perfectly.  We place a 
>>plastic
>>petri dish IN the liquid nitrogen, then you can set the OCT embedded 
>>spleen in the petri dish.  We have shattered spleen and if we do 
>>isopentane cooled by liquid nitrogen, or direct immersion into liquid 
>>nitrogen.  Just make sure the petri dish is supported so it does not
tip 
>>over in the Liq N2, and do not allow Liq N2 into the dish.  You need
to 
>>replenish the liq nitrogen as it is important to maintain the dish 
>>directly in the N2.  In other words, the dish goes canoeing in the Liq
N2.
>>
>>Good luck
>
>
>--


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