[Histonet] Re: [Histone) More on questions about restaining......

[Mildred Fail]

Don't Bleach  before restaining remove all the resin with xylene after removing the coverslip, rehydrate to water then repeat H&E only leave slides just a little longer even 20 yo slides will look like you stained them yesterday
>>> "Mindy Johnson" <m5johnso <@t> meded.ucsd.edu> 09/04/07 01:39PM >>>
Thanks for the emails about the tissue.. But I can disregard that as I have
found that our Medical Center has what I need!  But, I could still use some
trouble shooting help on my restaining issues...  Thanks everyone!

 

 Mindy A Johnson

SRA II

UCSD - School of Medicine

Medical Teaching Labs

Fax: 858-822-5250

 

From: Mindy Johnson [mailto:m5johnso <@t> meded.ucsd.edu] 
Sent: Tuesday, September 04, 2007 9:51 AM
To: 'histonet <@t> lists.utsouthwestern.edu' 
Subject: In need of tissue and questions about restaining......

 

Hello to all..

 

I am fairly new to histonet.  I am not a histologist by trade, but I am
doing a bunch of histology work at UCSD.  I have a few questions.  Does
anyone know where we can find some pathological tissue?  We need this for
students sets of pathology for our medical students.  It has become pretty
hard to find tissue.  I scoured the internet and found Biomax, but that has
been it.

 

Also, since I have no formal training (I wish I would have taken histology
courses in school) I need some advice on restaining some old tissue.  This
is for our student sets again.  The stain has faded significantly.  Most of
the tissue is at least 15 yrs old.  Some older.   Some a about 10 yrs old.
I have been soaking them in xylene for weeks and I can't seem to 1) either
get the resin off the slide or 2) I have no clue what this option is as they
have been soaking for so long and I do several changes!!  I tried restaining
some of they are pretty much ruined!  So, I have some others and could
really use some help!  If there is any.  Could the tissue be so old and used
so often under the microscope that it has degraded and is not useful
anymore?

 

This is what I have been doing:

 

Soak in xylene (do several changes)

Bleach with potassium permanganate and oxalic acid.

Restain with H&E protocol.

 

But, that wasn't working and was not consistent at all.  So I just tried
putting the slides back in the hematoxylin after soaking in xylene and I got
better results, with one type of tissue, but the older tissue didn't work so
well.

 

Thank you for all your help!!!

 

 Mindy A Johnson

SRA II

UCSD - School of Medicine

Medical Teaching Labs

Fax: 858-822-5250

 

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Rena Fail