Dear Pam,

> I am looking for help in establishing a reasonable turn around time for
>rapid processing of kidney biopsy specimens on transplant patients. Our
>clinicians  want "four hours from collection to microsc>
> One of our clinicians wants us to replace the formalin fixation with
>Zenkers (as they did in the good old days) to improve the quality, any
>comments on that suggestion?  Thanks in advance for your he>
>
Zenkers will take longer to fix than 10% formalin. Increase the time
in formalin as much as possible. Remember the average renal biopsy is
quite thin, so the biopsy will dehydrate and clear quite quickly. Wax
impregnation, I would expect to take longer than the clearing step.

Be careful of high temperatures during the processing and slide
drying stage. If sections are over cooked, especially since one would
expect them to be underfixed with formalin and therefore lack the
stability of thorough formalin crosslinking, nuclei will appear
opaque and lack the graininess of well fixed and processed chromatin.

A suggested protocol:
1.	Fix in formalin 1 hr
2.	75% alcohol 10 min
3.	90% alcohol 10 min
4.	Absolute alcohol 10 min x2
5.	Xylene 10 min x2
6.	Wax 10 min
7.	Wax 15 min
This will allow 30 min for embedding, sectioning and staining.
Cutting a bit fine but possible.

I hope this helps?
Regards, Tony
.

Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital
Sydney, AUSTRALIA

http://www2.one.net.au/~henwood
http://www.pathsearch.com/homepages/TonyHenwood/default.html