Re: [Histonet] Help with rodent tissue

From:Andrea Grantham



Cheryl,
I do a lot of rodent tissue here and even though I process it myself
occasionally there are some that are pretty dried out and need soaking. A few
months ago there was a discussion on histonet about putting glycerin in the
water that you soak your faced off blocks in. I tried this and it works
wonderfully. We don't measure out the glycerin although I think if you go back
in the histonet archives you will find the actual "recipe". We just put a small
amount of glycerin into a 500 ml bottle and fill 
it up with DI water - shake and
soak.
And I have noticed that my fingertips are now softer too!

Andi Grantham




At 07:42 AM 2/21/2008, Rene J Buesa wrote:
>Cheryl:
>   Rodent tissues are inherently lower in fat 
> and with less water in their fat tissue and 
> this is a "dangerous" combination when 
> processing because, as you point out, the 
> tissues you received are overprocessed so you 
> will have ptoblems sectioning, extending the 
> sections and holding them to your slides.
>   Try ALL the following steps:
>   1- prepare a solution of 0.2% (v/v) of fabric 
> softener and prepare a frozen block with it. 
> This frozen 0.2% fabric softerner will be used 
> to cold the blocks. Face them off and place 
> them face down over the frozen softener covered 
> with paper towels in a way that the blocks are 
> going to be in contact with the softener for at 
> least 30 minutes before trying to section.
>   2- increase the temperature of your water 
> bath at 50-52║C, close to the paraffin melting point WITHOUT reaching it.
>   3- add to your water bath 0.25 mL of liquid 
> detergent (but NOT dish washer). This step, and 
> the previous one are designed to lower the 
> water surface tension and facilitate the sections expansion.
>   4- add 1 mL of Elmer's (white) glue to the 
> water bath. The water will be slightly whitish 
> but this weak glue solution will help to hold the sections to the slides.
>
>   When you get to the area in the block that 
> you want to section, cool it with a ice cube 
> wrapped in a gauze and take the sections SLOWLY.
>
>   This combinaiton should help you, but all the steps together.
>   Good luck with your rodents' blocks!
>RenÚ J.
>
>Cheryl  wrote:
>
>
>
>
>
>
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.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
: (office:  AHSC 4212)          P.O. Box 245044                     :
: (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
: (FAX:  520-626-2097)          (email:  algranth@u.arizona.edu)       :
:...................................................................:
           http://www.cba.arizona.edu/histology-lab.html


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