RE: Rapid Processing of Kidney Biopsies

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From:Rob Geske <> (by way of histonet)
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Dear Pamela,

I am amazed and impressed when I see clinical professionals as yourself
meeting such demanding time constraints.  Working in research has many
rewards, and not having to deal with such ulcerogenic situations as you
describe is one of them.  I appreciate your concern about quality and think
that is a pertinent consideration.  I know at times it is difficult to find
time to "experiment" while doing the million and one things that need to be
done clinically, but would it be possible to obtain multiple renal biopsies
from the next autopsy and try various permutations of fixation and
processing (including any of the great suggestions you will get from fellow
netters)?  I would suggest that you get your pathologist involved so he/she
can see the relative results.  It is then up to them to choose what
schedule is within their comfort zone for making an appropriate/accurate
diagnosis and you should document their processing choice.  If "freshness"
of the autopsy specimen compared to the clinical sample is an issue (and it
might well be), you might consider an appropriate laboratory animal for the
survey.  I think this could provide a standard that you could compare your
future clinical samples against.

As far as Zenkers and "the good old days"...... it depends what the
pathologist is evaluating.  If the PAS is being done for basement
membranes, then Zenkers will be fine.  If it is being done to assess
glycogen content then Zenkers and other mercury fixatives are not
recommended.  Formalin is probably your best bet.  Besides, in a time
constrained system why would you want to add extra steps for removal of
mercury pigment?

Robert S. Geske
Research Associate
Center for Comparative Medicine and Department of Pediatrics
Baylor College of Medicine

-----Original Message-----
Sent:	Monday, November 29, 1999 9:37 AM
Subject:	Rapid Processing of Kidney Biopsies

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 microscopic slides
available for diagnosis." We have been trying to meet these expectations
but the quality is suffering, not to mention the level of stress in the
department. The major problem is the reduced processing times are impacting
the morphology. Is there anyone out providing this type of Rush service? If
so, would you be willing to share your procedure? Our current protocol is
to fix in 10% formalin, process on a short run, embed and to cut 20 serial
slides at 2 microns staining with H&E, PAS and trichrome. The PAS needs to
be available for interpretation with the H&E. The trichrome can be sent
later. I almost forget, this includes transport time from the University
campus to our consolidated lab (different campus, across the river) for
processing and the slides returned for interpretation. We usually received
 the specimens at 12:45 and have the slides available for return transport
at 3:45. Which reduces actual technical time to 3 hours.

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 help.

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