[Histonet] Fixation and Processing of Mouse Lactating Mammary Gland

[Catherine Simonson]

I think longer fixation AND processing.  Fix for at least 48 hours in NBF
and here is my processing schedule
70% for 45 min
70% for 45 min
95% for 45 min
95% for 45 min
100%  for 45 min
100% for 45 min
100% for 45 min
xylene  for 45 min
xylene for 45 min
xylene for 45 min
paraffin for 45 min
paraffin for 45 min
paraffin for 45 min
paraffin for 45 min

Good luck

On Tue, May 17, 2011 at 1:14 PM, Bell, Pat <Pat.Bell <@t> ucdenver.edu> wrote:

> Hello to all,
>
> I have some lactating mouse mammary tissue that was fixed in 10% formalin
> for 24 hours and then stored in 70% alcohol for 1 day before processing. The
> processing is as follows: 80%-30min, 95%-30min, 95%-40 min, 100% X3-30 min.
> each, xylene X3-40min. each, and paraffin X3-45min. each. The tissue if very
> mushy and I am wondering if the tissue was fixed long enough? The tissue is
> approximately 3mm in thickness but because of the milk in the tissue does it
> need to fix more or should I process longer?
>
> I have embedded the tissue and am wondering how I should go about
> reprocessing the tissue. Should I go back through xylene to alcohol and then
> fix for longer and then reprocess or can I melt the blocks and put in fix
> and then reprocess? I have heard both ways but want to do the method that
> will not comprise the IHC that needs to be done.
>
> Thank you so very much for all of your input and help.
> Pat
>
> Pat Bell HT(ASCP)
> University of Colorado, Denver
> Medical Oncology; MS 8117
> 12801 E 17th Ave.
> RC1-S, L18-8402C
> Aurora, Co. 80045
> 303-724-6077
> pat.bell <@t> ucdenver.edu<mailto:pat.bell <@t> ucdenver.edu>
>
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