[Histonet] RE: Histonet Digest, Vol 78, Issue 2
Mike Pence
mpence <@t> grhs.net
Mon May 3 08:18:39 CDT 2010
I had one of my techs come to me one day saying that she thought I
needed to look at her microtome because she had not been able to cut
anything on her microtome today. She thought something had gone wrong
with the tissue during processing. The other techs though were
sectioning fine. This tech has been with me for over 20 years. I went
over and looked at her microtome and told her that it might improve if
she tried a blade in the blade holder to start.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Liebig,
Tyler K.
Sent: Monday, May 03, 2010 7:32 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Histonet Digest, Vol 78, Issue 2
That is a funny story, I would like to see the slide that was cut from a
"blade holder section" with no blade in the clamp. I have heard it can
be done but never see it myself.
Though in old blade holders with loose clamps I have seen users get two
blades in there at once. That didn't section to well either.
Tyler
-----Original Message-----
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Sent: Sunday, May 02, 2010 10:04 AM
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Subject: Histonet Digest, Vol 78, Issue 2
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Today's Topics:
1. Revalidation (Tench, Bill)
2. RE-hydrating air dried slides for Pap stain (Jeffrey Silverman)
3. RE: H. D., Vol 78, Issue 1, Message: 7 (Keith)
4. RE: disposable blade holder (histotech <@t> imagesbyhopper.com)
5. RE: Slide and Block Storage (histotech <@t> imagesbyhopper.com)
----------------------------------------------------------------------
Message: 1
Date: Sat, 1 May 2010 10:37:02 -0700
From: "Tench, Bill" <Bill.Tench <@t> pph.org>
Subject: [Histonet] Revalidation
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <2820431BF953BB4DA3E9E1A5882265FD02863034 <@t> MAIL1.pph.local>
Content-Type: text/plain; charset=us-ascii
Kevin, trust your judgment. You have made significant changes in your
protocol. You need to revalidate. (and frankly, most changes in a
standard protocol would require revalidation).
Bill Tench
Associate Dir. Laboratory Services
Chief, Cytology Services
Palomar Medical Center
555 E. Valley Parkway
Escondido, California 92025
Bill.Tench <@t> pph.org
Voice: 760- 739-3037
Fax: 760-739-2604
mail2.pph.org made the following annotations
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Message: 2
Date: Sat, 1 May 2010 11:43:23 -0700 (PDT)
From: Jeffrey Silverman <pathmaster <@t> yahoo.com>
Subject: [Histonet] RE-hydrating air dried slides for Pap stain
To: vrodriguez10 <@t> gmail.com
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <789166.47922.qm <@t> web111111.mail.gq1.yahoo.com>
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Hi Valerie,
It sounds like you are performing a Pap stain using saline baths to
RE-hydrate air dried smears prepared at the bedside, I assume for?
staining with Diff Quik stains.
The rehydration ( not dehydration) takes place in the sodium chloride
baths and restores the morphology of the cells to something closer to a
wet fixed state. This takes place in 30 seconds.? You then must actually
fix the? now rehydrated cells in 95% alcohol. This step should be of
sufficient time to completely fix the cells as one would from the start,
up to 20 minutes is needed.
.
Next the cells need to be rehydrated again, just placed in a water
rinse, before staining with hematoxylin and proceeding with the Pap
cytoplasmic stains.
Awareness of these steps should obviate any problems.
Finally, I wouldn't judge the success or failure of the stain by the
color of the slide since different tissue fluid backgrounds and amounts
of lysed blood might stain differently.? Check the slide
microscoipically to assess the stain.
Jeff Silverman
------------------------------
Message: 3
Date: Sat, 1 May 2010 12:05:58 -0700
From: "Keith" <mikoff <@t> pacbell.net>
Subject: [Histonet] RE: H. D., Vol 78, Issue 1, Message: 7
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <NLENIJCPKPOMIABJIDGIIEEBGGAB.mikoff <@t> pacbell.net>
Content-Type: text/plain; charset="us-ascii"
Here's a copy of a procedure that provided consistently adequate results
from my old one-man lab histo days...
For your case, perhaps the slides in question were fixed differently or
improperly(?) I would pull the questionable slides, delicately rehydrate
them, run a control slide, and reprocess them with the following
procedure. In this procedure, the staining agents, dehydrants, and
clearing agents and can be modified to your facility and reagent list.
Keith M. Mikoff, HTL/HT (ASCP)
PAPANICOLAOU STAIN (PAP)
PURPOSE: The following staining procedure is to be performed
only
on spray fixed slides. It is a routine stain used for any
cytol-
ogy (i.e., PAP smears, FNA procedures, or specimens of a fluid
nature where cytology is required).
PROCEDURE:
1. Dehydrate slides in 2 changes of isopropyl alcohol, 5
minutes each.
2. Rinse in tap water for two minutes.
3. Stain in Harris' Hematoxylin for 2 minutes.
4. Rinse in fresh tap water for 3 minutes.
5. Place in ammonia water for 1 minute.
6. Rinse in fresh tap water for 1 minute.
7. Dehydrate in 3 changes of Isopropyl alcohol, 2 minutes
each.
8. Stain in O-G 6 for one minute.
9. Dehydrate in Isopropyl alcohol for 2 changes, 2 minutes
each.
10. Stain in EA-50 for 4 minutes.
11. Place in Isopropyl alcohol, 3 changes, at 1 1/2 minutes
each
12. To clear the slides, place in 3 changes of Histoclear,
for 2
minutes each, and coverslip.
------------------------------
Message: 4
Date: Sun, 2 May 2010 09:01:04 -0400
From: <histotech <@t> imagesbyhopper.com>
Subject: RE: [Histonet] disposable blade holder
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <1729E60B0B6D456BB67E686C815CC071 <@t> hopperPC>
Content-Type: text/plain; charset="US-ASCII"
As a comical sidelight to the last sentence that Tyler wrote ...
I have a pathologist who was very upset that our cryostat didn't seem to
be working properly. I rushed up to the frozen room to assist and he
said, "well, nevermind, I got a slide and diagnosis." I looked in the
cryostat and said to him "you actually got a section and were able to
render a diagnosis?" "yes" he said. "WOW!, you must be the ONLY
pathologist who is capable of sectioning tissue without a knife ..."
;o) He sectioned the tissue and truly had no idea there wasn't a knife
in the blade holder!
:o)
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Liebig,
Tyler K.
Sent: Friday, April 30, 2010 3:41 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] disposable blade holder
Hello Lucy,
In regards to your trouble finding an HM310 blade holder we do have one
in our refurbished pool that I could have sent to you. It is used but in
working order so we can let you have it at no charge. If you let me
know where to ship it I will have it sent.
Hope that helps, It's hard to cut without a blade holder.
Regards,
Tyler Liebig
Product Manager
Histology Instrumentation and IHC
Anatomical Pathology
Thermo Fisher Scientific
46360 Fremont Blvd
Fremont, CA 94538
USA
Office: (510) 979-5000
Mobile: (510) 299-1751
Fax: (510) 979-5239
tyler.liebig <@t> thermofisher.com<mailto:tyler.liebig <@t> thermofisher.com>
www.thermo.com<http://www.thermo.com>
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Message: 5
Date: Sun, 2 May 2010 09:04:16 -0400
From: <histotech <@t> imagesbyhopper.com>
Subject: RE: [Histonet] Slide and Block Storage
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <7180DF002734414D8FEEE9A4A4600940 <@t> hopperPC>
Content-Type: text/plain; charset="US-ASCII"
In our facility, we maintain both the blocks and slides.
We also store older blocks/slides in a climate controled an off-site
facility.
Michelle
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Elizabeth Kronenberger
Sent: Friday, April 30, 2010 11:00 AM
To: 'Nails, Felton'; 'Daniel Adams'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Slide and Block Storage
Clients fax orders for deepers and special stains which we perform and
send them out with their next package for them to read and report.
-----Original Message-----
From: Nails, Felton [mailto:flnails <@t> texaschildrens.org]
Sent: Friday, April 30, 2010 10:48 AM
To: 'Elizabeth Kronenberger'; 'Daniel Adams';
histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Slide and Block Storage
So what if the reporting company need to order special stains, it adds
too much time having to request the blocks or additional unstained
slides.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Elizabeth Kronenberger
Sent: Friday, April 30, 2010 9:40 AM
To: 'Daniel Adams'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Slide and Block Storage
If our lab does slide preps for other clients (Technical component
only), we hold the blocks at our facility. The slides are read and
reported by the client, they hold the slides.
Liz
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Daniel
Adams
Sent: Friday, April 30, 2010 12:41 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Slide and Block Storage
My laboratory provides professional services to some clients and
technical services to others. The question has recently come up, who
should store blocks and slides when the pathologist and histology
laboratory are different companies?
- Dan
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