[Histonet] mouse heart chatter upon sectioning...
Rene J Buesa
rjbuesa <@t> yahoo.com
Thu Jun 5 07:37:37 CDT 2008
Mouse tissues are specially low in fat and prone to become brittle with too much ethanol or xylene. I think you should cut the dehydration times in half and use isopropanol (ISO) instead of ethanol.
After the ISO place the hearts in a mixture of ISO and mineral oil (M) 5:1 for 1 hour at 50ºC, another hour in a ISO:M mixture 2:1 at 50ºC; another hour in pure M at 50ºC, another hour in a 1:1 mixture of M and paraffin at 50ºC, and complete your infiltration with paraffin as usual.
This method has been a modification by Maxim Peshkov of my mineral oil method, Give it a try. It is now standard procedure in a veterinary lab in the UK.
"Gaupp, Dina D " <dgaupp <@t> tulane.edu> wrote:
Whenever I process (whole)mouse hearts, the tissue chatters upon sectioning. The tissue is usually fixed overnight in 10%nbf at 4C before processed. I tried different processing times. The last time I processed hearts, I processed them for 5 hours total no vaccum. Should I decrease the processing time even lower than that? I am afraid of underprocessing. I have to section the entire heart from top to bottom, so I really need some advice.
(open system) Processor Set Up:
1 - fix (if tissue fixed overnight, skip #1 & #2 stations)
2 - fix
3 - 70%
4 - 80%
5 & 6 - 95%
7 & 8 - 100%
9 & 10- Xylene Sub. - Clear-rite
11 & 12-Paraffin stations
dgaupp <@t> tulane.edu
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