From:Bruce Gapinski

No, maybe I'm extra cautious. I refer you to an article in CAP Today 1198
Vol. #12 No.1 pg. 59-61 section - Summing Up by Patrick C. Roche, Ph.D. He
recommends cutting IHC fresh. Because I'd rather mount the control on the
same slide as the "unknown" tissue, I find this convenient. Why spend time
doing a study to find out when pre-cut tissues are too old? 
Bruce Gapinski HT(ASCP) 

		-----Original Message-----
		From:	Richard Cartun []
		Sent:	Thursday, October 04, 2001 6:05 AM
		Subject:	RE: IHC

		Hi Bruce:

		I find it amazing that you have to cut all tissues fresh and
store your control blocks in the freezer.  We pre-cut all of our control
tissues and have observed little, if any, diminution of immunoreactivity
over time.  Yes, there are a few controls (especially nuclear antigens)
where we may only cut enough controls for only 1-2 months.  Is there
something unusual about your fixation or tissue processing that creates this
problem for you?

		R. Cartun

		>>> Bruce Gapinski  10/02/01
11:21AM >>>
		I've had great experience w/ BioGenex reagents. What they
don't have you may
		find w/ Vector or Dako. Controls are specified by the
manufacturer, and you
		should be working closely w/ your Pathologist(s). We used to
send out cases
		to an IHC lab, then we tried the same procedure in-house w/
our reagents.
		The Pathologist should make the final "call" on all the
stains. You can buy
		control slides from BioGenex (I'll bet Dako sell them too).
That would be a
		good start, I think. 
		QA - all IHC stains in our lab must be run w/ a negative
(both control and
		patient). Save reagents by mounting the control on the same
slide as the
		"unknown" slide or patient slide. One last touch is to cut
all tissue fresh
		each time. Stored control slides may stain lighter. With
HER-2 neu this is
		critical, as the staining intensity is what the pathologist
is looking for.
		We store all our IHC control blocks in the freezer, so they
are ready when
		we need them. 
		Happy trails. Bruce Gapinski HT(ASCP)

				-----Original Message-----
				From:	Ryan, Genoula K Ms USAMH
				Sent:	Tuesday, October 02, 2001 5:07 AM
				To:	'Bruce Gapinski'
				Subject:	RE: IHC

				What kind of antibodies/controls did you
use?  How about
		QA/QC?  Is there a
				lot more one would have to do?  

				-----Original Message-----
				From: Bruce Gapinski
				Sent: Monday, October 01, 2001 4:40 PM
				To: 'Ryan, Genoula K Ms USAMH'
				Subject: RE: IHC

				I use to do IHC by hand. Low budget; Just
find an old gram
		stain rack. Etch
				around the tissue w/ a scribe and put your
antibodies on.
		Use a timer and
				that's about all you'll need. Be careful not
to FLOOD the
		slide w/ expensive
				reagents. Respectfully,
				Bruce Gapinski HT(ASCP)

						-----Original Message-----
						From:	Ryan, Genoula K Ms
						Sent:	Monday, October 01,
2001 3:36 AM
						To:	'HistoNet Server'
						Subject:	IHC

						To HistoNetters,
						Our new Chief of Pathology
would like us to
		do a few
				immunos.  We are not
						set up to do any.  Is there
a cheap way to
		do immunos?  We
				don't want to
						purchase a Ventanna or
anything fancy.  Our
		workload is
				small and he only
						wants the basics (i.e.,
S100, CK, ER/PR).
		Any suggestions
				on how to get
						this up and running will be
		appreciated.  Also if
				there are any good
						reference books for IHC.
Thanks in advance,
						Genie K. Ryan,
						Histology Supervisor
						DSN: 371-2653

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