[Histonet] Fixation and Processing of Mouse Lactating Mammary Gland

[Bell, Pat]

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>