[Histonet] Mammary Tissue Processing & Sectioning Troubles

Logan, Shannon Shannon.Logan at bellin.org
Wed Sep 27 15:31:51 CDT 2017


Hi Andrea,
Tissue that is not fixed properly will
be mushy and difficult to section, just as you described.  Was there a reason NOT to send it in NBF for storage?

I think the main problem is with the storage in 70%. Or there is a small possibility the 14 hr schedule did not remove all of the water from
the tissue. However, I tend to doubt the tissue was fixed in the first place.
If they said the tissue was fixed for 2 days in NBF I would question that as well.
If it was never fixed or not fixed long enough, then storage in 70% would
be detrimental to proper fixation in the 14 hr schedule.



Shannon H. Logan BS, HTL (ASCP)

Bellin Health
Pathology Dept.
744 South Webster Ave.
Green Bay, WI  54305
920-433-3653

From: Andrea Calhoun via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Wednesday, September 27, 2017 2:26 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Mammary Tissue Processing & Sectioning Troubles

Hi All!

I'm working with primate mammary tissue that was given to us from a pathology group off campus. They say the tissue was fixed in NBF for at least two days, and sent to us in 70% EtOH, where it has sat for a couple months. We are treating these samples as we would with human tissue. After grossing, processing (14hr schedule), and sectioning (4-5um, high-profile blade)), I find the tissue is difficult/ near impossible to section. Even after leaving the blocks on ice for 30+min, the tissue continuously mushes against the blade. If I do get it to cut, multiple knife marks develop quickly and I find I am going through blades like crazy. I am staying superficial incase the fix didn't penetrate deep into the tissue. So far I've only processed a few samples from a larger group to resolve these issues before processing the remainder.

Does anyone have any tips for processing or sectioning? Besides grossing the tissue into smaller pieces, would it help to re-fix them overnight or an additional day? How would you tell if the issue is from fixation or insufficient clearing/infiltration of paraffin?

Any information would be appreciated!

Andrea Calhoun B.S., CEMT
Research Assistant 2, Schedin Lab
Dept. of Cell, Developmental, & Cancer Biology
Oregon Health & Sciences University
calhouan at ohsu.edu<mailto:calhouan at ohsu.edu> , RJH-5350

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