[Histonet] RE: processing and staining preserved specimen

[Schade, Adelle]

Thank you Tom and Pat-
Here are the biopsy sizes: 

I have my classes experimenting with specimen size to try to find what will work best for our situation.  My 1st block class has 2mm biopsy cores, 2nd block has 4mm biopsy cores and 3rd block has 6mm biopsy cores- each about 2- 3 mm thick.

I will try some specimen altering the times as Tom suggested to see what we get- Thanks again
Adelle

-----Original Message-----
From: Truscott, Tom [mailto:ttruscot <@t> vetmed.wsu.edu] 
Sent: Friday, October 17, 2014 1:32 PM
To: Schade, Adelle; histonet <@t> lists.utsouthwestern.edu
Subject: RE: processing and staining preserved specimen

Hi Adelle, Biopsy sizes will matter, but you might shorten some of these times, but be sure to add at least one more Histoclear to make sure there is no alcohol in it and at least one more paraffin to make sure there is no histoclear carryover.If you go back to 70% at any time you will have to start over, so try to get a janitor or someone to start early or a coach to finish late. Make sure you keep things safe, because of all the flammables. Tom T 

-----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 10:09 AM
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