[Histonet] processing and staining preserved specimen

[Schade, Adelle]

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