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

b427297 <@t> aol.com b427297 <@t> aol.com
Fri Aug 3 16:02:53 CDT 2012

This is speculation on my part, but since your brain is about 60% lipid, you are depleting it by storage in alcohol. Most antibodies in my experience work well after 48 hours in formalin, so why don't you just let them stay in formalin over the weekend and diminish time in alcohol? 

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On Aug 3, 2012, at 2:30 PM, Jamie E Erickson <jamie.erickson <@t> abbott.com> wrote:

> 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 
> (3X).
> 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. 
> 132-137(6).
> 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 
> sections
> Thanks,
> Jamie
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