[Histonet] RE: processing and staining preserved specimen

[Marcum, Pamela A]

First of all THANK YOU for getting our field into a high school and teaching the kids.  This is a very good thing for all of us in the field.  

Could you give us an idea of the size of the specimen you are attempting to process?  A great deal depends on the tissue size as to how long it can be left in each reagent and paraffin.  I realize your limitation in time and did hand processing of monkey and rat brains as well as other tissue, for years.  No automated system was available and in research no money for one at the time.  I would be more than happy to share. 

Have you checked with TJ and maybe HUP to see if they old equipment that is still good from a research lab that might be available to be donated?  (Yes, I am from Philly)   

Pam Marcum
UAMS

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Schade, Adelle
Sent: Friday, October 17, 2014 12:09 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] processing and staining preserved specimen

Hello everyone,
I have been following and learning from this listserv for the past two years, I have enjoyed all of the advice this forum presents.  This is my first inquiry for the group.  I am a high school Anatomy and Physiology teacher and a grad student at Thomas Jefferson (Philadelphia, PA).  In our research lab, we process, section, embed and image for all research projects.  I was lucky enough to acquire some used histology equipment for my high school lab this school year.  I am teaching the students about histology and the processing.  We have an embedding station, microtome, staining dishes and a good microscope to image.  We do not have an automatic tissue processor so we are processing manually (I prepare all solutions for the students).  We are taking biopsy samples from our preserved mink that we are dissecting at the moment.  I have processed the tissue using the following method and have pretty good results, not great.  (we are doing a basic H&E stain to see the cells of the tissue).  If we do not complete the  manual processing method and need to leave school for the day, I move the cassettes into 70% EtOH until the next morning when we continue the process.  I am wondering if anyone has manually processed preserved specimen tissue in the past and has any advice/ protocol that you would suggest?  This is what I am using:

Processing Method

1.  70% EtOH:  1 hour
2.  80%  EtOH:  1 hour
3.  90%  EtOH:  1 hour
4.  95% EtOH:  2 hours
5.  100% EtOH:  2 hours
6.  Histoclear II:  2 hours
7.  Paraffin:  2 hours

Also- I will eventually be looking for grants to try to purchase an automatic tissue processor- this would alleviate the time issues we are facing.  Does anyone know of grants that would focus on the educational aspect of histology?
Thank you for your time and have a great weekend!
Adelle Schade
Graduate student:  Jefferson School of Biomedical Science


Ms. Adelle L. Schade, B.S., M.Ed.
Anatomy and Physiology
Conrad Weiser High School
44 Big Spring Rd.
Robesonia, PA  19551
a_schade <@t> conradweiser.org



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