[Histonet] Microchatter in mouse brain(isopropanol-storage solution)

Jamie E Erickson jamie.erickson <@t> abbott.com
Fri Aug 3 14:30:18 CDT 2012

Hi All,
              I  Hope someone can provide me with a little insight on a 
problem I'm having. 
I am do IHC on mouse brains that I fix for 8-12 hrs (trimmed after 2-3 
hrs) in 10% NBF and processed overnight , embedded next day.
 My processor holds samples after fixation (8Hr)  in 70% ETOH until it can 
be completed in the AM. My stations are  25min/ station + 1 Hr in paraffin 
If the study comes down on a Friday  I hold samples  in Isopropanol 70% at 
4 degrees C  for the weekend this was in the journal of histotechnology  
Journal of Histotechnology, Volume 34, Number 3, September 2011 , pp. 
I'm noticing the brains are way to dry and I see  a lot of microchatter on 
my sections. The IHC is good but these microchartter is difficult to deal 
with. I notice if  I hold my block on wet ice for 1 hr  they are not 
swollen, if this was a normal mouse brain w/o isopropanol it would be 
swollen up by then, so it is dry...

So what is a guy to do? Do I reduce the Conc of isopropanol to say 40%..
I was running these samples right away. Fixed 6 hrs and processed  right 
away  and I  didn't have this problem but  coming back late at night to 
embed these studies was getting old....

So anyone have similar issues  I'd like to hear how you resolved them.
Extending fixation is possible but I'm not sure the effect on the IHC, 
I've already done 4 studies this way and I had to change but I need better 



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