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

From:Gayle Callis

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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