Re: Rush Kidney Process

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From:"Gregory Lloyd" <> (by way of histonet)
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Pamela wrote:

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."

Hi Pamela

We have developed a process where we can have the biopsy slides on the
pathologists desk in about 3 hours (this includes dividing times for EM, and
Immunofluorescence). I'm not sure how it will fit into your lab but there
are two caveats.  One, there is a slight to moderate shrinkage of the
glomerulus.  Two, this requires a very large amount of tech time to perform.

There is a lab requirement that the biopsy must be considered stat, and that
the time of receipt must be after 2 o'clock in the afternoon.  If both of
these conditions are present then a tech is pulled from their bench (a
rotational transplant tech) and others fill in the gap so the transplant
tech can do the biopsy. If one of the conditions is not met/necessary (the
path's decision) then it is processed on a four hour schedule VIP processor
for that same day or next day.

The process is very easy and requires a microwave, beakers and other common
lab equipment.  The major skill is the experience to determine processed end
points of the various solutions.

The needle core biopsy is fixed in formalin in a 60 degree oven for 20
minutes, then transferred to 70% alcohol (at 60 degrees) for 10 minutes.
Two solutions of absolute alcohol are heated in succession for 34 sec until
the temp reached 62degrees +\- 3 degrees.  The biopsy is placed in each for
3 minutes in a 60 degree oven.  Xylene is then heated to 43 degrees and the
tissue is processed for 3 minutes in this solution.  Then the biopsy is
transferred to two molten paraffin containers at 75 degrees for 3 minutes
each.  Embed and then cut.  Our pathologists read only the H&E's in rush
cases the special stains are held until the next working day.

Before your safety rep jumps all over you and says "potentially explosive"
we do this in a well ventilated room and cover the solutions when they are
being heated and used.  No open flames are permitted in the lab as a
requirement and small volumes (150 ml) are being used.

Sorry I didn't intend this to be so lengthy, there is more.  If you would
like to see the entire procedure you can e-mail me back and I can forward it
directly to you.

Cameron Lloyd
Edmonton Alberta.

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