Re: [Histonet] FW: Microtome at home

[Mark Tarango via Histonet]

I had heard that CLIA was relaxing things and is not requiring a new # to
work from home right now.  Best to check on the regulatory but FFPE isn't
typically infectious.  The ideal spot would in the garage and not the
kitchen though.

On Thu, Apr 16, 2020 at 3:47 PM Roxana Robinson via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> I do not agree with  this in our current situation or actually any
> situation.
> There are quidelines in place with CLIA, OHSA  and CAP for protecting not
> only the patient but also the employee.  Whether research or not.
>
>
> Roxana Robinson
>
> > On Apr 16, 2020, at 4:58 PM, Patsy Ruegg via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > I agree with this point and as far as clocking in and out, I would
> think you could work out something like getting paid piece mill, perhaps
> charge per slide or block cut, that way you could do it on your own time
> and not have to clock in.
> >
> >
> > Patsy Ruegg, HT(ASCP)QIHC
> > Ruegg IHC Consulting
> > 40864 E Arkansas Ave
> > Bennett, CO 80102
> > H 303-644-4538
> > C 720-281-5406
> > prueg...@hotmail.com
> >
> >
> > 
> > From: Joseph Saby 
> > Sent: Thursday, April 16, 2020 8:03 AM
> > To: Porter, Amy ; Porter, Amy via Histonet <
> histonet@lists.utsouthwestern.edu>; histonet@lists.utsouthwestern.edu <
> histonet@lists.utsouthwestern.edu>; Steven Crochiere  >
> > Subject: Re: [Histonet] FW: Microtome at home
> >
> >
> > You will need to make sure all pertinent SOPs and EOPs are followed, as
> well as all safety guidelines/protocols. Just because it is not human
> tissue doesn't mean that it can't have its share of nasties.
> > Joe Saby
> >
> > Sent from Yahoo Mail on Android
> >
> > On Thu, Apr 16, 2020 at 8:21 AM, Porter, Amy via Histonet<
> histonet@lists.utsouthwestern.edu> wrote:   Make sure of insurance
> coverage and safety for the employee and that they are covered in case of
> injury - are they still clocking in and out in some fashion. just
> thinking in a bigger box.
> >
> > 
> > From: Steven Crochiere via Histonet 
> > Sent: Thursday, April 16, 2020 6:36 AM
> > To: histonet@lists.utsouthwestern.edu  >
> > Subject: [Histonet] FW: Microtome at home
> >
> > Jaime,
> >
> > I don't see a problem with a research setting. If it was patient care,
> CLIA would need to inspect the set up in the person home. The same goes for
> our pathologists who read slide at home.
> >
> > Steve
> >
> > -Original Message-
> > From: raestask via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> > Sent: Wednesday, April 15, 2020 7:51 PM
> > To: Jamie Watson ;
> Histonet@lists.utsouthwestern.edu
> > Subject: Re: [Histonet] Microtome at home
> >
> > I wouldn't think there would be any problem.Rae Staskiewicz HT(ASCP)Sent
> from my Verizon, Samsung Galaxy smartphone
> >  Original message From: Jamie Watson via Histonet <
> histonet@lists.utsouthwestern.edu> Date: 4/15/20  6:44 PM  (GMT-06:00)
> To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microtome at
> home Hello all,Our pathologist has come up with the idea of sending a
> microtome and waterbath home to someone that cannot come to work due to
> COVID 19.  We are a research lab and work with mouse and rat tissue.  Does
> anyone know of any issues with doing this?  I have never heard of anyone
> cutting slides at home other than someone with a private business.Thank
> you.Jamie___Histonet mailing
> listHistonet@lists.utsouthwestern.eduhttp://
> lists.utsouthwestern.edu/mailman/listinfo/histonet
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> >
> >
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Re: [Histonet] pan-TRK or NTRK antibody

[Mark Tarango via Histonet]

 While we're on the topic of NTRK, does anyone have a positive case that
they could share?  I'm working up the FISH and have probes for NTRK1, 2 &
3.  Any pan-NTRK positive IHC case would be great for detecting by FISH too.

thanks!

Mark Tarango

On Wed, Sep 11, 2019 at 11:23 AM Piche, Jessica via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Good Afternoon,
>
> Where are people buying the pan-TRK or NTRK antibody from? Pre-dilute
> preferred.  Thank you in advance and have a great day!
>
> Jessica Piche, HT(ASCP)
> Waterbury Hosptial
> Waterbury, CT 06708
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Re: [Histonet] FISH for breast core (Her2)

[Mark Tarango via Histonet]

 Hello Taganrog,

If you are having issues with high autoflorescence due to prolonged
formalin fixation (7 days), it might help to do a few minutes extra
protease digestion (5+ minutes) during tissue pretreatment.  Alternately or
in conjunction, I would suggest re-applying HER2 probe to the same slide
that had already been pretreated and hybridized.  By doing this a few times
(even up to four tries), you will likely get FISH signals that you can see
and enumerate.

Good luck,

Mark Tarango

On Tue, Jun 11, 2019 at 11:33 AM Пешков Максим via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

>
> Dear colleagues!
> Can anyone help? We are very appreciatated any suggestions, tips and hacks
> for Her2 FISH onto breast trepine fixed in 10% NBF. It was here for more
> than 7 days, then processed as usual into wax. IHC for Her2 has value as 2+.
> Sincerely,
> Maxim Peshkov,
> Russia,
> Taganrog.
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Re: [Histonet] Histonet Digest, Vol 184, Issue 1

[Mark Tarango via Histonet]

How about passing on to the department that could fix the issue?  Where's
the empowering innovation?

On Fri, Mar 1, 2019 at 10:35 AM Jordan, Kelley via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> More bad press in histonet...not sure if we should pass it on to
> Marketing??
>
> @Will, Who as Teri Blaud at   Holy Redeemer Hospital ? Teri is always
> always bashing us.
> Kelley Jordan
>
> Strategic Account Manager - SC, NC, TN and KY
>
>
> A Member of the Roche Group
>
> Ventana Medical Systems
>
> Mobile:  803.504.1135
> Customer/Technical Support: 1.800.227.2155
>
> kelley.jor...@roche.com
>
> www.ventana.com
>
>
>
> Empowering | Innovation
>
>
> Confidentiality Note: This message is intended only for the use of the
> named recipient(s) and may contain confidential and/or proprietary
> information. If you are not the intended recipient, please contact the
> sender and delete this message. Any unauthorized use of the information
> contained in this message is prohibited.
>
> histonet 
>
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Re: [Histonet] Another Dispenser Failure

[Mark Tarango via Histonet]

I hope everyone is using on-slide controls :-)

On Thu, Feb 28, 2019 at 11:52 AM Terri Braud via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Another one!!!
> Our Her2 antibody dispenser failed, LOT #E22628
> This one "supposedly" FDA approved.
> Roche, why do you continue to lie to consumers of your product?  You claim
> you've "fixed" the problem but your Ventana dispensers DON'T WORK!
> This is patient care!  Why don't you care about the customers and patients
> you are supposed to be serving
> Shame on you.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> -Original Message-
> From: histonet-requ...@lists.utsouthwestern.edu [mailto:
> histonet-requ...@lists.utsouthwestern.edu]
> Sent: Thursday, February 28, 2019 1:00 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: Histonet Digest, Vol 183, Issue 23
>
>
> Today's Topics:
>1. H Staining question (Charles Riley)
>2. Re: H Staining question (Jay Lundgren)
>3. FYI- Roche Ventana users (Cassie P. Davis)
> Message: 3
> Date: Thu, 28 Feb 2019 15:23:16 +
> From: "Cassie P. Davis" 
> To: histonet 
> Subject: [Histonet] FYI- Roche Ventana users
> Message-ID: 
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi Histoland,
> I am biting my tongue HARD and just letting you know so it doesn't
> happend to you. I just got off the phone with Roche here is the heads-up.
> If one of their anitbody dispensers fails DO NOT put the antibody in one
> of their prep kits, as soon as you do they consider it off label use.
> Call customer service immediately and have them overnight a replacement!
> Cassandra Davis
> Histology Technician
> AP Laboratory
> 302-575-8095
> Email:  cda...@che-east.org
>
>
>
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[Histonet] NTRK positive tissue

[Mark Tarango via Histonet]

Would anyone have NTRK1, NTRK2, or NTRK3 positive tissues?  We are trying
to validate break-apart FISH probes for these gene translocations but don't
have any positive tissue.  If you have something positive by IHC, I would
be very happy to try FISHing it.

thank you

Mark Tarango
CellNetix Pathology
Seattle, WA
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Re: [Histonet] FISH question

[Mark Tarango via Histonet]

Hi Gudrun,

Are you sure you have digested long enough with pepsin?  If the tissue is
not well digested you will see background.  We use sodium thiocyanate for
pretreatment reagent, not citric buffer.  These are my first thoughts.

Mark

On Tue, Feb 19, 2019 at 9:18 AM Gudrun Lang via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Dear histonetters!
>
> I have difficulties with my FISH preparation on FFPET. I struggle with
> massive background. It looks like a thick fluorescent film.
>
> The signals can't be seen because of the background. Even the nuclei are
> hard to see.
>
> The background is within the tissue but also surrounds it. Therefore it
> must
> be directly on the glass slide.
>
>
>
> The slide is clear after deparaffination and after pretreatment with citric
> buffer and pepsin. After the pepsin the slides are rinsed in 2xSSC, then
> 50%-70%-96%-100% ethanol (p.a.).
>
> And then the slides are airdried.
>
> On the dry slides foggy streams appear. The slides become turbid. When I
> rinse them again in graded ethanols it becomes better but still a little
> turbid.
>
> After hybridisation and stringent washing the slides are air-dried again
> and
> coverslipped with Dapi.
>
> When looking at the slides in the fluorescence microscope the trouble
> arises.
>
>
>
> My assumption is, that there is a remnant of the salt of the SSC buffer.
> How
> can I inhibit this deposit? Can I replace the buffer with water without any
> harm to the tissue?
>
> Or ist there a different cause for the turbidiy?
>
> I use fresh reagenses from xylene to buffer and ethanol.
>
> Any hints are welcome.
>
>
>
> Thanks in advance
>
> Gudrun Lang
>
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Re: [Histonet] pathologic staging

[Mark Tarango via Histonet]

The pathologist should be responsible for pathologic staging on excisional
specimens.  The surgeon (sometimes oncologist) does the clinical staging.
At least that is what happens at our local tumor board meeting.

Mark

On Mon, Nov 12, 2018 at 9:05 AM Eck, Allison via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Good morning histonet
> I have a question that is more for a pathologist but I am hoping some of
> you can help.  When staging cancer cases (namely breast), there are 2
> stagings, pathologic and clinical  As per the AJCC, the pathologic staging
> includes  information defined at surgery and clinical staging is determined
> by using information prior to surgery or neoadjuvant therapy.  My question
> is: Are pathologists responsible for staging both the clinical and the
> pathologic stage?  If not, do you know who does the clinical staging?
>
> Thanks
> Allison
>
> Allison Eck, HTL(ASCP)cm,QLS, AHI(AMT),CEAS1
> Lead Tech Histology
> Doylestown Hospital
> 595 W State St
> Doylestown, PA 18901
> 215-345-2264
> a...@dh.org
>
>
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Re: [Histonet] Unstained slides - how long are they good for?

[Mark Tarango via Histonet]

Hi Everyone!

I have seen unstained slides save a patient from re-biopsy many times.
Usually it will be a case where a patient has a known diagnosis, like lung
cancer.  In these types of cases after diagnosis molecular testing (and IHC
for PD-L1) is usually ordered.  There have been countless times that I can
recall where a few unstained slides on a biopsy with scant tumor was able
to get us results for PD-L1, ALK FISH, and ROS1 FISH.  Often in these types
of a cases a touch prep can be used for Next Generation Sequencing or PCR
testing like EGFR or BRAF, allowing for the full panel of molecular tests
to be performed.  For cases that are small specimens I would prefer to have
unstained slides to fall back on for patient convenience, client
satisfaction, and quicker TAT of molecular testing.

Re-biopsy and re-diagnosing the new sample costs money to the patient and
payers and having some unstained slides can often save those costs
providing more value to the original biopsy.  Sometimes when we try to save
money in the lab it can result in more money being spent on healthcare
overall. It is true that some antigens become more difficult to stain over
time and storage is an important consideration.  Limiting the production of
unstained slides to small and scant needle may make storage more practical.

Just some more things to consider.

Sincerely,

Mark Tarango


On Thu, Aug 16, 2018 at 4:48 PM, P Sicurello via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hello My Fellow Histologists,
>
> Happy Friday Eve.
>
> The question has come up..  How long are *unstained* slides good for?
> Not for H but tests like IHC and molecular testing.  These slides have
> been cut, stored at room temperature, not sealed in anyway, and kept in a
> cardboard box.
>
> Please let me know what your opinions are and what your retention policy is
> concerning *unstained* slides.
>
> Thanks oodles.
>
> Sincerely,
>
> Paula Sicurello, HTL (ASCP)CM
>
> Histotechnology Specialist
>
> UC San Diego Health
>
> 200 Arbor Drive
>
> San Diego, CA 92103
>
> (P): 619-543-2872
>
>
>
> *Confidentiality Notice*: The information transmitted in this e-mail is
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Re: [Histonet] Breast Her2Neu IHC vs FISH

[Mark Tarango via Histonet]

We add a comment when the specimen handling is outside of 2013 CAP/ASCO
guidelines for breast or 2016 CAP/ASCO/ASCP guidelines for HER2 testing in
GEA cancers.  These guidelines and the data supplements have some good
information.  We usually have found that decal doesn't affect the IHC much
if not excessive (but still add a comment) and we do attempt performing
FISH and have to rely internal controls (normal 2-signal pattern on
non-neoplastic cells) to know that the FISH reaction is appropriate.  FISH
working will depend on time in formalin before decal and time in decal.
This changes from specimen to specimen, so it may be difficult to validate
the full range of specimen variability.  If you do FISH, the best trick is
to re-apply the probe on the same slide that was already denatured and
hybridized the previous day and run it through denaturation and
hybridization again.  This will bring out the signals in many cases.

Since FISH has internal controls (normal 2-signal pattern in non-neoplastic
cells), it can help give some confidence to a negative HER2 IHC result when
specimen handling was compromised.  I believe that is why the CAP question
below says to perform "confirmatory analysis by in-situ hybridization".

These are some of our comments
Decal:

This assay has not been validated on decalcified tissues.  Results should
be interpreted with caution given the possibility of false negativity on
decalcified specimens.  (we don't add this comment if the result is
positive).

Formalin fixation:
The specimen did not meet optimal formalin fixation guidelines (fixed for
xxx hours).  This can play a role in loss of FISH signals.  Repeat testing
on an appropriately fixed specimen is recommended, if clinically feasible.

Fixation and decal:
The specimen did not meet optimal formalin fixation guidelines (fixed for
xxx hours and xxx minutes).  Both factors may play a role in weak or
missing FISH signals.  Repeat testing on a non-decalcified and properly
fixed specimen is recommended, if clinically feasible.

There is a CAP question that asks about some of those factors.  Not sure if
you are CAP-accredited or not.

**REVISED** 07/28/2015
ANP.22983 HER2; ER/PgR - Fixation Phase I
If the laboratory assesses HER2 protein over-expression by
immunohistochemistry, HER2
(ERBB2) gene amplification by in situ hybridization, or
estrogen/progesterone receptor
expression by immunohistochemistry, there is a written procedure to ensure
appropriate
specimen fixation time.

Anatomic Pathology Checklist 08.17.2016
NOTE: Specimens subject to these tests should be fixed in 10% neutral
buffered formalin for
at least six hours and up to 72 hours. The volume of formalin should be at
least 10 times the
volume of the specimen. Decalcification solutions with strong acids should
not be used. For
cases with negative HER2 results by IHC that were fixed outside these
limits, consideration
should be given to performing confirmatory analysis by in-situ
hybridization.
Laboratories must communicate the following fixation guidelines to clinical
services:
1. Specimens should be immersed in fixative within one hour of the biopsy
or resection
2. If delivery of a resection specimen to the pathology department is
delayed (e.g.
specimens from remote sites), the tumor should be bisected prior to
immersion in
fixative. In such cases, it is important that the surgeon ensure that the
identity of the
resection margins is retained in the bisected specimen; alternatively, the
margins
may be separately submitted.
3. The time of removal of the tissue and the time of immersion of the
tissue in fixative
should be recorded and submitted to the laboratory
Communication may be through memoranda, website, phone, face-to-face
meetings, or other
means. The laboratory should consider monitoring compliance and contacting
clients when these
guidelines are not met.
If specimens are fixed in a medium other than 10% neutral buffered
formalin, the
laboratory must perform a validation study showing that results are
concordant with
results from formalin-fixed tissues.
Laboratories testing specimens obtained from another institution should
have a policy that
addresses time of fixation. Information on time of fixation may be obtained
by appropriate
questions on the laboratory’s requisition form.
Reports should qualify any negative results for specimens not meeting the
above guidelines.
Reports containing ER, PgR or HER2 results for their predictive
characteristics must specify the
type of fixative used and the cold ischemia time. In addition, any
treatment that may potentially
alter immunoreactivity, such as decalcification, must be included.


Mark Tarango


On Wed, Apr 19, 2017 at 12:29 PM, Jason McGough via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Does anyone know or can you point me in the right direction to some
> literature about how to properly test for Her2Neu (IHC vs. FISH) on breast
> tumors if the cold ischemia ti

Re: [Histonet] Her2 IHC

[Mark Tarango via Histonet]

My lab does not have a written policy on this yet but are discussing it.
If the patient has a high grade tumor and HER2 IHC is 1+ (or even 0+), some
of our pathologists will reflex to FISH.  The oncologists will request it
sometimes too, more often when ER and PR are both negative.  We have found
several cases to be positive by FISH that were 0+ or 1+ by IHC.  I can't
say anything about response to HER2 therapy in this group of patients just
that they met the criteria for positive by FISH.

Mark T.
Cellnetix
Seattle, Wa

On Thu, Nov 3, 2016 at 9:12 AM, Algeo, Lacie A via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Are any labs reflexing 1+ for FISH?
> Thanks :)
> Lacie
>
> Lacie Algeo, HTL (ASCP) MBCM
> Histology Supervisor
> Providence Sacred Heart Medical Center Laboratory
> 101 W 8th Avenue
> L-2
> Spokane, WA 99204
> 509-474-4418
> FAX 509-474-2052
> lacie.al...@providence.org
>
>
> This message is intended for the sole use of the addressee, and may
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Re: [Histonet] Slide scribe

[Mark Tarango via Histonet]

You could try these from Ted Pella:
http://www.tedpella.com/glasswar_html/scriber.htm


On Wed, Mar 2, 2016 at 10:19 AM, Cartun, Richard via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Years ago, I obtained a metal scribe for etching glass slides, but I can't
> remember which Histology company I got it from.  Any suggestions?  Thank
> you.
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & The Martin M. Berman, MD Immunopathology &
> Morphologic Proteomics Laboratory
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 972-1596
> (860) 545-2204 Fax
>
>
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[Histonet] MET amplification - need help

[Mark Tarango via Histonet]

Hello everyone,

I am working a validation for MET amplification by FISH and am having a
hard time finding positive cases.  Would anyone have any to share or
trade?  I have access to all kinds of cases from both IHC and molecular for
exchange.

thanks

*Mark Tarango*
Lead Molecular Technologist - FISH

CellNetix Pathology & Laboratories
1124 Columbia Street, Suite 200
Seattle, WA 98104
PH: 206-576-6526
Fax: 206-215-5946.
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Re: [Histonet] Freeze Spray Not Sold as Case

[Mark Tarango via Histonet]

I used to work in a lab that used compressed air held upside down as freeze
spray.  Worked the same for me and might be cheaper.



On Wed, Aug 19, 2015 at 6:53 PM, ian bernard via Histonet 
histonet@lists.utsouthwestern.edu wrote:

 Fellow Histonetters: I'm looking for a source to purchase Freeze-Spray used
 during the frozen procedure.  Our current resource sells Freeze Spray in a
 case of 6 cans. I would like to purchase as an individual can owing to our
 workload rather than a case.



 Any references?



 Ian Bernard



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Re: [Histonet] Positive Control for IF?

[Mark Tarango]

It is April 1st, isn't it?

On Wed, Apr 1, 2015 at 4:12 PM, Paula Sicurello pat...@gmail.com wrote:

 Good Afternoon Netters,

 Since we went down the path of hot dogs and Slim Jim's as positive controls
 for FFPE stains, I was wondering.

 Is there a source, like a hot dog or piece of steak, that could be a
 positive control for immunofluorescence C4d?

 What I'd really like to find is some food or food product (our positive
 patient biopsies for the frozen IF are teeny-tiny) that is positive for the
 whole host of IF stains:  IgG, IgA, IgM, Kappa, Lambda, you get the
 picture.

 Please send your suggestions my way.

 Thanks in advance,

 Paula  :-)
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Re: [Histonet] ALK IHC

[Mark Tarango]

Did you ever run ALK IHC on lung cancer cases or have they always used FISH?

thanks

Mark Tarango

On Fri, Oct 24, 2014 at 12:34 PM, Joelle Weaver joellewea...@hotmail.com
wrote:

 Recently all the pathologists I work with prefer the ALK FISH.


 Joelle Weaver MAOM, HTL (ASCP) QIHC





  Date: Thu, 23 Oct 2014 13:41:15 -0700
  From: marktara...@gmail.com
  To: Histonet@lists.utsouthwestern.edu
  CC:
  Subject: [Histonet] ALK IHC

 
  Does anyone stain lung cancer specimens for ALK using IHC? If not, any
  opinions?
 
  thanks
 
  Mark
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[Histonet] ALK IHC

[Mark Tarango]

Does anyone stain lung cancer specimens for ALK using IHC?  If not, any
opinions?

thanks

Mark
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Re: [Histonet] NCCI interpretation for ISH coding

[Mark Tarango]

My understanding was that this is just for Medicare patients...



On Mon, Jan 6, 2014 at 12:35 PM, Johns, Jill jjoh...@cpallab.com wrote:

 The following was taken from the NCCI manual, Chapter 10, effective 1/1/14:

 9. The unit of service for in situ hybridization reported as CPT codes
 88365, 88367, or 88368 is each probe staining procedure per specimen. If a
 single probe staining procedure for one or more probes is performed on
 multiple blocks from a surgical specimen, multiple slides from a cytologic
 specimen, or multiple slides from a hematologic specimen, only one unit of
 service may be reported for each separate specimen. Physicians should not
 report more than one unit of service for CPT codes 88365, 88367, or 88368
 per specimen for a probe staining procedure even if it contains multiple
 separately interpretable probes.

 I'm wondering how other labs are interpreting this, because I'm not sure
 how we continue to offer FISH testing and not go in the hole financially,
 if we can only bill for ONE probe on ONE specimen, regardless of the actual
 number of interpretable probes (Example: currently, for a HER2 FISH test,
 we bill for 2 units of service--1 for the HER2 probe and 1 for the CEP17
 probe--both of which have to be enumerated by a professional and a ratio
 generated). Any insight from others would be greatly appreciatedThanks!


 Jill A. Johns, MT(ASCP)SH, QCym, CCy
 Manager of Molecular Pathology
 Central PA Alliance Laboratory (CPAL)
 1803 Mt. Rose Ave., Suite C3/C4
 York,  PA17403
 phone: (717) 851-4320
 fax:  (717) 851-1450
 email:  jjoh...@cpallab.commailto:jjoh...@cpallab.com


 
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Re: [Histonet] DNA Extraction

[Mark Tarango]

Hi Elizabeth,

Room temp is fine for FFPE tissue for DNA extraction.  We do it this way
every day!

thanks

Mark


On Tue, Jul 30, 2013 at 12:02 PM, Elizabeth Cameron 
elizabeth.came...@jax.org wrote:

 We are preparing paraffin slides for DNA extraction, and I am not sure if
 once cut they should be kept cold before the extraction is done.  We are
 not doing the extraction, just the prep.  If anyone else is doing this, I
 would greatly appreciate some input.
 Thank you,
 Elizabeth

 The information in this email, including attachments, may be confidential
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Re: [Histonet] Biocare Medical Mach 4

[Mark Tarango]

Hi Kris,

The polymer backbone for this detection kit is conjugated with anti-rabbit
antibodies only (AP Polymer reagent).  The specific probe is a rabbit
anti-mouse antibody in the AP probe reagent.  That is why you only need to
apply the AP probe when your primary antibody is from mouse but not when
it's rabbit.

Mark


On Tue, Jun 11, 2013 at 12:05 PM, Kalleberg, Kristopher 
kristopher.kalleb...@unilever.com wrote:

 Hello All,

 Does anyone have any knowledge of the Biocare Medical Mach 4 AP detection
 system.  How does this system exactly work.  Does it use a dextran backbone
 with 2ndary antibodies attached like other systems and then the polymer
 binds to that?  Their datasheets simply state that it uses a specific probe
 and then the polymer binds to that.  I am very pleased with the results I
 have achieved with this system but am a little confused on the technique of
 labeling.  Any knowledge on this topic would be greatly appreciated.  And
 as always, thanks in advance.

 Kris
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Re: [Histonet] IHC on non-charged slides

[Mark Tarango]

Yes, if the stain can wait get some mount quick from Newcomer.  There is a
good chance the tissue will fall off if you proceed with the tissue on
regular slides.  I would wait until I get the mount quick.

Everyone else reading this should order a tube now for emergencies.  You
will find yourself in this position at some point and if you have the stuff
on hand, it's no problem.

I just used it today to transfer cells from a cytocentrifuge slide to a
charged slide for FISH.  It saved the patient a re-biopsy.

Mark


On Thu, May 16, 2013 at 2:00 PM, Debra Siena dsi...@statlab.com wrote:

 Michael

 There is a procedure where you can transfer the sections to a charges
 slide with a product called mount quick. You can check with newcomer supply
 they sell it. I know of several labs that have used it on these occasions
 and it works well.

 Sent from my iPhone

 On May 16, 2013, at 4:50 PM, Dessoye, Michael J 
 mjdess...@commonwealthhealth.net wrote:

  I'm hoping Histonet can come through for me!  I have an unusual case
 that I need to run IHC on.  Antibody is Pan Keratin Cocktail from Cell
 Marque with recommended protocol on a Benchmark Ultra with i-view
 detection.  Only problem is, I only have two slides and they are not
 charged.  The tissues have been air-dried but not baked.  I cannot obtain
 more slides.
 
  I'm wondering if there's any kind of pre-treatment I could try to try to
 help the tissue stay on.  We have run slides like this in the past with
 mixed results.  Most of the time, the tissue washes off.
 
  Any tips or tricks that might help out in this situation?
 
 
 
  Michael J. Dessoye, M.S. | Histology Supervisor | Wilkes-Barre General
 Hospital | An Affiliate of Commonwealth Health |
 mjdess...@commonwealthhealth.net | 575 N. River Street | Wilkes Barre, PA
 18764 | Tel: 570-552-1432 | Fax: 570-552-1526
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Re: [Histonet] RE: Image analysis

[Mark Tarango]

I'd like to add that we use the Aperio system with Ventana's antibody and
it works well.  It took a lot of work tweaking of the algorithm to get the
software to score accurately using the 4B5 clone but that was the stain our
pathologists were already used to.

Mark


On Tue, Apr 23, 2013 at 1:50 PM, Elizabeth Chlipala l...@premierlab.comwrote:

 Pat

 There is a list of 510K cleared algorithms on the DPA website - Here is
 the link.


 https://digitalpathologyassociation.org/_data/files/DPA_Regulatory-FDA-510k_list.pdf

 Liz

 Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
 Laboratory Manager
 Premier Laboratory, LLC
 PO Box 18592
 Boulder, CO 80308
 Work (303) 682-3949
 Fax (303) 682-9060
 Cell (303) 881-0763
 l...@premierlab.com
 www.premierlab.com

 Ship to address:

 1567 Skyway Drive, Unit E
 Longmont, CO 80504



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Zeitlow
 Sent: Tuesday, April 23, 2013 1:34 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Image analysis

 Looking for opinions on digital image analyzers and software... want to do
 quantitative image analysis for ER/PR and HER2 IHC.


 Thanks!

 Greg Z




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Re: [Histonet] Formalin Neutralizer

[Mark Tarango]

Our safety person claims no test for formalin is accurate after the
addition of sodium sulfite.  I had suggested some kind of testing after I
was surprised by the strength of the fumes when someone was pouring the
treated formalin waste down the drain.  I wish I knew more about it.




On Thu, Apr 11, 2013 at 6:26 PM, Cristi Rigazio cls71...@gmail.com wrote:

 We have a formaldehyde test kit.  It's a dip stick type test.

 Sent from my iPhone

 On Apr 11, 2013, at 5:31 PM, Mark Tarango marktara...@gmail.com wrote:

 Can I ask how you test before dumping?

 Thanks

 Mark
 On Apr 11, 2013 6:21 AM, Cristi Rigazio cls71...@gmail.com wrote:

 We neutralize ours and have no problems with it.  I am not sure how much
 you use, so I will say it is easiest in smaller batches as you do have to
 shake it up to make sure it dissolves.  We purchase ours from BBC
 Biochemical for a very reasonable price and then test before dumping.  We
 have doing this for four years with no issues.
 Thanks,
 Cristi

 Sent from my iPhone

 On Apr 11, 2013, at 6:04 AM, Bustamante, Lin lbustama...@cvm.tamu.edu
 wrote:

  We are looking into the option of neutralizing our formalin waste
 instead of having it to be picked up.
  If you use Formalin Neutralizer, do you have any  pro/con about this
 product?
  Thank you very much.
 
  Lin S. Bustamante, B.S., H.T.(ASCP)
  VIBS Histology Laboratory Supervisor
  College Of Veterinary Medicine
  Texas AM University
  College Station, Texas 77843-4458
  Phone: (979) 845-3177
  Fax: (979) 458-3499
 
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Re: [Histonet] RE: Out of my comfort zone...

[Mark Tarango]

We use a heat block during digestion.  There is less chance of
contamination with a heat block than with a water bath.

... and no we don't use antigen retrieval solution for this!  We use a
Qiagen kit too.

Mark


On Wed, Apr 3, 2013 at 9:11 AM, Helen Fedor hfe...@jhmi.edu wrote:

 We have been using store bought gallons of distilled water in our water
 baths. This water has been boiled so enzyme activity should be absent.


 Helen

 410.614.1660

 http://tmalab.jhmi.edu/
 http://prostatebiorepository.org/



 Helen

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sue Hunter
 Sent: Wednesday, April 03, 2013 10:55 AM
 To: Sarah Dysart; histonet@lists.utsouthwestern.edu
 Subject: [Histonet] RE: Out of my comfort zone...

 Hello
 Just wanted to add one more thing - we actually use a dedicated pyrex dish
 (maybe 6x10 inches) for our water bath for RNA sections.  We use warm tap
 water, but you can put it in the microwave for a short bit if it needs to
 be warmer.  You can spray the dish with RNAse away and wipe before filling
 with water.
 Sue

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
 Sent: Tuesday, April 02, 2013 5:36 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Out of my comfort zone...

 So...I have been asked to do some micro-dissection on some slides and then
 do downstream RT/PCR on them.  My molecular knowledge doesn't go much out
 of the world of IHC so...here is my question...

 Has anyone ever substituted Citrate Buffer pH6 (or whatever HIER solution
 you are using) for proteinase K for use in RNA isolation and then later
 PCR?  Does this work?  The main question is will the HIER step take off the
 formalin linkage from the nucleic acids, or just the protein?

 One last thing is what else goes into these solutions other than Citrate
 Buffer and Tween?  I haven't made it up in forever, I have just been
 ordering it from companies...I know...lazy...

 Thanks

 Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna
 Therapeutics
 2150 Woodward Street
 Suite 100
 Austin, Texas  78744
 (512)901-0900 ext. 6912

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Re: [Histonet] Ventana Labels

[Mark Tarango]

Hi Matthew,

We use them and they work fine.  They are much cheaper too.  We've had them
for about 3 years.  I initially had some concerns about the DAB being
absorbed and staining the labels on some cases.  Our safety person told me
that it's not a safety concern and it's safe to touch the label after it's
stained brown by DAB.  This doesn't happen on every slide but is a random
thing.  It might have something to do with how well it was affixed to the
slide.

The labels don't affect the staining in my experience.

Mark

On Mon, Feb 25, 2013 at 11:33 AM, Matthew Roark mro...@sfmc.net wrote:

 Hello all,

 I am getting ready to try some slide labels as an alternative to the
 Ventana
 labels.  I was wondering if anyone else is using them or have tried them in
 the past.

 They do not have that plastic fold over flap and are suppose to be quite
 economical, though I have not got a quote yet.

 They are called FloProTek (http://www.easternlabsvc.com/floprotek.php) -
 Mercedes Medical Supply sent me the trial roll.

 Thanks!


 Matthew Roark- HT/HTL(ASCP)CM
 Histology Specialist
 Saint Francis Medical Center
 211 Saint Francis Drive
 Cape Girardeau, MO 63703
 573-331-5267
 mro...@sfmc.net
 http://www.sfmc.net





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Re: [Histonet] FISH baking and dewaxing

[Mark Tarango]

The package insert for PathVysion (HER2 FISH) says to bake overnight at 56
degrees C.  We never do this and our signals are clear, bright, and
punctate.  Many of the slides are just air-dried.  Some are baked for about
20 minutes and I don't notice any difference between them.  I wonder if I
would see a difference with overnight baking...

The aging step we use for cytology preps to be FISHed is 2 minutes in 2x
SCC at 73 degrees C before protease digestion.  I didn't know extended
baking was considered an aging step for tissue sections to be FISHed.
 Thanks Gudrun!

On Fri, Feb 15, 2013 at 7:20 AM, Gudrun Lang gu.l...@gmx.at wrote:

 We see better results with FISH-slides baked over night. This is a
 phenomenon called aging, known from cytogenetics.
 It seems, that the time of air-drying and heat leads to a better access of
 the probes to the dna.
 Therefor we let them also in the oven over the weekend.
 Gudrun


 -Ursprüngliche Nachricht-
 Von: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Houston,
 Ronald
 Gesendet: Donnerstag, 14. Februar 2013 20:08
 An: histonet@lists.utsouthwestern.edu
 Betreff: [Histonet] FISH baking and dewaxing

 Can someone please explain why paraffin-embedded slides for FISH need to be
 baked for so long and have extensive dewaxing in xylenes and subsequent
 alcohol pretreatment?
 Our Molecular group bake for 1 hr at 65C, and then treat sections in 3
 changes of xylene 15 min each, and 100% ethanol 15 minutes each x2 before
 air-drying. I have asked them why so long and get the standard response --
 that's the way we always do it!

 Thanks
 Ronnie

 Ronnie Houston, MS HT(ASCP)QIHC
 Anatomic Pathology Manager
 ChildLab, a Division of Nationwide Children's Hospital www.childlab.com

 700 Children's Drive
 Columbus, OH 43205
 (P) 614-722-5450
 (F) 614-722-2899
 ronald.hous...@nationwidechildrens.orgmailto:
 ronald.houston@nationwidechild
 rens.org
 www.NationwideChildrens.orghttp://www.nationwidechildrens.org/

 One person with passion is better than forty people merely interested.
 ~ E.M. Forster

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Re: [Histonet] Her2neu controls

[Mark Tarango]

Hi Jim,

We run the control slide that Ventana sends (although not the best control)
but we add a section from our tissue control block before staining.  The
tissue control block contains a 0+, weakly staining 1+, and a 3+.  We use
the ventana/tissue control slide as our batch control.  This slide must be
QC'd by a pathologist before any HER2 IHC is sent to pathologists for
scoring.  In addition, each patient slide has a section of the tissue
control.  We use a weakly staining 1+ in the tissue control to monitor any
slight change in the staining.  If the 1+ suddenly is a 0+, then we have a
problem.  If you were just running a 3+ control, you probably wouldn't
notice the staining difference in the control.

Mark T.

On Mon, Feb 11, 2013 at 2:53 PM, Vickroy, Jim vickroy@mhsil.com wrote:

 When we order the antibody from Ventana they include some control slides
 with tissues that are 0, 1+, 2+, and 3+.   Currently the lab where we have
 sent our Her2neu's to be done only uses a 3+ control.  I realize that a
 control that had several levels of staining would be ideal but I am trying
 to find out how other diagnostic labs are handling their controls for
 Her2neu.
 Thanks




 James Vickroy BS, HT(ASCP)

 Surgical  and Autopsy Pathology Technical Supervisor
 Memorial Medical Center
 217-788-4046


 
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Re: [Histonet] Process and hold?

[Mark Tarango]

I second that!

On Tue, Feb 12, 2013 at 11:12 AM, Robert Schoonhoven 
robert_schoonho...@yahoo.com wrote:

 All you need to do is have them drain the chamber and place the cassettes
 on a nonabsorbent surface and allow them to cool to room temp.   On Monday
 morning have the techs put the into the cassette storage on your embedding
 center and they should be ready to embed witin a few minutes.  As the
 tissues are processed through paraffin they are protected and will not
 dry out.


 Robert Schoonhoven, HT/HTL (ASCP)


 
 From: Tom McNemar tmcne...@lmhealth.org
 To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
 
 Sent: Tuesday, February 12, 2013 1:56 PM
 Subject: [Histonet] Process and hold?

 Hello all,

 I was wondering about processing breast specimens (needle cores) on
 Fridays.

 We have asked our radiology department to try to avoid scheduling these
 breast biopsies on Fridays since we do not work weekends and are concerned
 about the extended time in formalin.

 I am thinking that we can run these specimens on a second processor over
 Friday night and have someone from the clinical lab come up  and drain the
 paraffin.  The tissues would then sit in a warm moist retort until Monday
 morning when they would be embedded and cut.  I think the specimens would
 be fine.  Processing would be complete at that point and they would hold in
 the unopened retort chamber.

 Our alternative is to have someone come in every Saturday morning just to
 remove and embed these specimens.



 Tom McNemar, HT(ASCP)
 Histology Co-ordinator
 Licking Memorial Health Systems
 (740) 348-4163
 (740) 348-4166
 tmcne...@lmhealth.orgmailto:tmcne...@lmhealth.org
 www.LMHealth.org
 file:///C:\Documents%20and%20Settings\TMCNEMAR\Application%20Data\Microsoft\Signatures\
 www.LMHealth.org

 
 This e-mail, including attachments, is intended for the sole use of the
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 use of the contents of this e-mail and attachments is prohibited. If you
 have received this in error, please advise the sender by reply e-mail and
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Re: [Histonet] High complexity test

[Mark Tarango]

.
 (ii) Score 3. Specialized scientific and technical knowledge is
   essential to perform preanalytic, analytic or postanalytic phases of
 the
   testing.
 (2) Training and experience.
 (i) Score 1. (A) Minimal training is required for preanalytic,
   analytic and postanalytic phases of the testing process; and
 (B) Limited experience is required to perform the test.
 (ii) Score 3. (A) Specialized training is essential to perform the
   preanalytic, analytic or postanalytic testing process; or
 (B) Substantial experience may be necessary for analytic test
   performance.
 (3) Reagents and materials preparation.
 (i) Score 1. (A) Reagents and materials are generally stable and
   reliable; and
 (B) Reagents and materials are prepackaged, or premeasured, or
   Require no special handling, precautions or storage conditions.
 (ii) Score 3. (A) Reagents and materials may be labile and may
   require special handling to assure reliability; or
 (B) Reagents and materials preparation may include manual steps such
   as gravimetric or volumetric measurements.
 (4) Characteristics of operational steps. (i) Score 1. Operational
   steps are either automatically executed (such as pipetting,
 temperature
   monitoring, or timing of steps), or are easily controlled.
 (ii) Score 3. Operational steps in the testing process require close
   monitoring or control, and may require special specimen preparation,
   precise temperature control or timing of procedural steps, accurate
   pipetting, or extensive calculations.
 (5) Calibration, quality control, and proficiency testing materials.
 (i) Score 1. (A) Calibration materials are stable and readily
   available;
 (B) Quality control materials are stable and readily available; and
 (C) External proficiency testing materials, when available, are
   stable.
 (ii) Score 3. (A) Calibration materials, if available, may be
   labile;
 (B) Quality control materials may be labile, or not available; or
 (C) External proficiency testing materials, if available, may be
   labile.
 (6) Test system troubleshooting and equipment maintenance.
 (i) Score 1. (A) Test system troubleshooting is automatic or self-
   correcting, or clearly described or requires minimal judgment; and
 (B) Equipment maintenance is provided by the manufacturer, is seldom
   needed, or can easily be performed.
 (ii) Score 3. (A) Troubleshooting is not automatic and requires
   decision-making and direct intervention to resolve most problems; or
 (B) Maintenance requires special knowledge, skills, and abilities.
 (7) Interpretation and judgment. (i) Score 1. (A) Minimal
   interpretation and judgment are required to perform preanalytic,
   analytic and postanalytic processes; and
 (B) Resolution of problems requires limited independent
   interpretation and judgment; and
 (ii) Score 3. (A) Extensive independent interpretation and judgment
   are required to perform the preanalytic, analytic or postanalytic
   processes; and
 (B) Resolution of problems requires extensive interpretation and
   judgment.


 Tim Morken
 Supervisor, Electron Microscopy/Neuromuscular Special Studies
 Department of Pathology
 UC San Francisco Medical Center





 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
 Sent: Wednesday, February 06, 2013 2:07 PM
 To: Jesus Ellin
 Cc: histonet@lists.utsouthwestern.edu
 Subject: Re: [Histonet] High complexity test

 Just to clarify, this is not my interpretation.  This is what CAP will
 tell you when you give them a call.

 Mark

 On Wed, Feb 6, 2013 at 1:07 PM, Jesus Ellin jel...@yumaregional.org
 wrote:

  I would say this is high complexoty testing and the tech performing
  this has to have knowledge of the process and troubleshooting in case
  there is issues with the results.  I do not agree with the
  interpretation some people give,, but this is based on individual
  institutions
 
  Sent from my iPad
 
  On Feb 6, 2013, at 2:05 PM, Rene J Buesa rjbu...@yahoo.com wrote:
 
   This issue has been discussed at length recently (please go to
   HistoNet
  files).
   The complexity does not deals with the actual test but with the
  ability of the technician to go above and beyond the robotic tasks
  but also able to think and apply knowledge when something goes wrong.
   Sometimes dismissal of complexity is rooted on the desire in
   management
  to pay less for tasks that require a higher licensure grade.
   René J.
  
   From: Sara Baldwin/mhhcc.org sbald...@mhhcc.org
   To: histonet@lists.utsouthwestern.edu
   Sent: Wednesday, February 6, 2013 2:54 PM
   Subject: [Histonet] High complexity test
  
   Hi histonetters
   Is ventana Ultra IHC only doing antibodies no FISH

Re: [Histonet] ER/PR and Her2 image analysis

[Mark Tarango]

Hi Cheryl,

We use the Aperio system for HER2 scoring.  Our lab manager put a cytotech
in charge of validating the digital reading and her next project is ER and
PR.  For HER2 IHC, the software is initially set for Dako's Hercept test.
 We don't use Dako, we use Ventana staining platform and the 4B5 clone for
HER2 IHC.  She had to play with the software until it was scoring
cases appropriately, but it works great now.

The initial slide scanner we got from them was a lemon.  They sent another
and it was a lemon too.  The third instrument (which is a newer model)
usually works without any problems.

The system helps with FISH too.  The pathologist can mark the area of
interest for FISH on the digital slide image.  This is great for my lab
because we have pathologists that are not here at our main lab site.  This
allows me to mark the area and start FISH even though the IHC or HE might
be off-site.

Mark T.

On Tue, Feb 5, 2013 at 7:07 PM, Cheryl tkngfl...@yahoo.com wrote:

 Hey- Help!

 Will ya'll share what image analysis software/hardware you're using?  Is
 anyone doing full digital quatifying analysis of ER, PR and Her2 ?


 Thanks!


 Cheryl Kerry, HT(ASCP), AP Manager
 Pathology Group of Louisiana
 cke...@pathgroupla.com

 (LOOK GUYS!!  No digest :) )
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Re: [Histonet] Re: HER-2

[Mark Tarango]

HER2 FISH really is considered the best method.  It's the only method that
has actually been tied directly to patient outcome.  The other methods are
expert consensus based.  That being said, there are some cases that are
equivocal by FISH.

Sometimes we have to ignore the CEP-17 (green) signals and report the HER2
status based only the HER2 copy number.  If the HER2/CEP17 ratio is
equivocal but the average HER2 count is over 6, we add a comment explaining
that it can be considered positive.  If it's between 4-6, we say
it's equivocal and usually suggest repeating on the excision.  If it's
under 4, we say that it can be considered to be negative.  We can resolve
most cases this way.  In rare instances we have sent out to another lab to
perform alternative FISH probes to enumerate for chromosome 17 (probes
SMS/RARA to Phenopath).  We have not found this to be especially useful.

If a case is 3+ by IHC but for some reason the oncologist wants FISH to
confirm, the patient will not be treated if FISH is negative.  There could
be other reasons besides gene amplification for accumulation or expression
of the HER2 protein.  The drug is only proven to work when the gene is
amplified and you test that by FISH.

PCR has it's own problems too.  The biggest thing, I think, is that you
dilute your HER2 score by including normal tissue during macro or
micro-dissection.  This does not work well for small foci of tumor.  Many
stomach and esophagus biopsies contain only small amounts of tumor.

Mark

On Wed, Feb 6, 2013 at 7:03 AM, Bob Richmond rsrichm...@gmail.com wrote:

 Mark Tarango notes:

 Many pathologists, if they have any doubt about the score will just say
 that it is 2+ so that its gets HER2 by FISH which is considered the best
 method for determining HER2 status.

 On one busy pathology service I worked in 2004-2006 we were quite
 explicit about overcalling HER-2 by IHC 2+. If we had any doubts at
 all, we sent out the FISH. The cost is trivial compared to the cost of
 treating a woman with trastuzumab (Herceptin).

 It's important to understand that IHC and FISH do not measure the same
 thing - IHC is looking at the excessive amount of the gene product,
 while FISH is looking at amplification of the gene itself. Neither is
 necessarily the best method. Some oncologists want both methods done
 on all cases.

 PCR further complicates the situation. I think some reference labs now
 consider this the preferred HER-2 method for adenocarcinomas of the
 stomach and esophagus.

 Bob Richmond
 Samurai Pathologist
 Maryville TN

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Re: [Histonet] High complexity test

[Mark Tarango]

If you are just staining the slides and not reading them, then you are NOT
performing high complexity testing.  The person who reads the slide is
doing the high complexity part.

Mark

On Wed, Feb 6, 2013 at 11:54 AM, Sara Baldwin/mhhcc.org
sbald...@mhhcc.orgwrote:

 Hi histonetters
 Is ventana Ultra IHC only doing antibodies no FISH or CISH is this
 considered High complexity testing?  We are doing ER/PR and some others.

 Thanks
 Histology/Cytology Supervisor
 S. Kathy Baldwin, SCT (ASCP)
 Memorial Hospital and Health Care Center
 sbald...@mhhcc.org
 Ph 812-996-0210, 0216, Fax 812-996-0232,
 Pager 812-481-0897, Cell 812-887-3357
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Re: [Histonet] High complexity test

[Mark Tarango]

Just to clarify, this is not my interpretation.  This is what CAP will tell
you when you give them a call.

Mark

On Wed, Feb 6, 2013 at 1:07 PM, Jesus Ellin jel...@yumaregional.org wrote:

 I would say this is high complexoty testing and the tech performing this
 has to have knowledge of the process and troubleshooting in case there is
 issues with the results.  I do not agree with the interpretation some
 people give,, but this is based on individual institutions

 Sent from my iPad

 On Feb 6, 2013, at 2:05 PM, Rene J Buesa rjbu...@yahoo.com wrote:

  This issue has been discussed at length recently (please go to HistoNet
 files).
  The complexity does not deals with the actual test but with the
 ability of the technician to go above and beyond the robotic tasks but
 also able to think and apply knowledge when something goes wrong.
  Sometimes dismissal of complexity is rooted on the desire in management
 to pay less for tasks that require a higher licensure grade.
  René J.
 
  From: Sara Baldwin/mhhcc.org sbald...@mhhcc.org
  To: histonet@lists.utsouthwestern.edu
  Sent: Wednesday, February 6, 2013 2:54 PM
  Subject: [Histonet] High complexity test
 
  Hi histonetters
  Is ventana Ultra IHC only doing antibodies no FISH or CISH is this
 considered High complexity testing?  We are doing ER/PR and some others.
 
  Thanks
  Histology/Cytology Supervisor
  S. Kathy Baldwin, SCT (ASCP)
  Memorial Hospital and Health Care Center
  sbald...@mhhcc.org
  Ph 812-996-0210, 0216, Fax 812-996-0232,
  Pager 812-481-0897, Cell 812-887-3357
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Re: [Histonet] Joe Nocito's request -- oh my!

[Mark Tarango]

Am I the only one who thinks its funny Joe the Toe did an unsubscribe
e-mail and Cheryl Kerry copied an entire digest to reply.  Oh histonet pet
peeves... haha

On Mon, Feb 4, 2013 at 1:05 PM, Cheryl tkngfl...@yahoo.com wrote:

 Oh no, Joe!  Say it ain't so!!  Please tell us this is just to take off
 your military address and you aren't leaving us forever...

 Cheryl


 Cheryl Kerry, HT(ASCP)
 Full Staff Inc.
 Staffing the AP Lab by helping one GREAT Tech at a time.
 281.852.9457 Office
 800.756.3309 Phone  Fax
 ad...@fullstaff.org

 Sign up for the FREE newsletter AP News--updates, tricks of the trade and
 current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe'
 request to apn...@fullstaff.org. Please include your name and specialty
 in the body of the email.

 --- On Mon, 2/4/13, histonet-requ...@lists.utsouthwestern.edu 
 histonet-requ...@lists.utsouthwestern.edu wrote:
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Re: [Histonet] HER-2

[Mark Tarango]

Many pathologists, if they have any doubt about the score will just say
that it is 2+ so that its gets HER2 by FISH which is considered the best
method for determining HER2 status.  Even if by the scoring criteria it is
a 1+ but the intensity is a little stronger than normal (but maybe
basolateral or not complete staining) they just go with 2+.  Sometimes its
high grade 1+ and it doesn't quite meet the 2+ staining criteria and they
call it 2+ too.  If these things are happening enough it could mean calling
more 2+ cases.  They don't always follow the scoring criteria to the letter.

We use digital image analysis for HER2 IHC scoring and the computer is
pretty right on (matches with FISH), but sometimes the pathologist will
change the score in the report to 2+ even though it's a 1+ by the computer.
 There is one pathologist in particular who doesn't believe in the computer
reading the HER2 score and is trying very hard to find cases that are
positive by FISH but that the computer is calling the IHC 1+.  He also
requests FISH on some 3+ cases to try and find any over-calling by the
computer.  The thought of a woman getting a toxic drug needlessly really
bothers him.

So without Wilson posting more info there's not much help that I think
anyone can offer.  This is stain that has to be validated
more extensively than others, so the protocol shouldn't just be tweaked.
 How do the controls look?  Was there any lot to lot variation?  Lots of
questions..

Mark

On Tue, Feb 5, 2013 at 12:49 PM, Rene J Buesa rjbu...@yahoo.com wrote:

 Yes, that is sometimes a common occurrence amongst pathologists, BUT those
 differences have to be solved in conference before issuing the report.
 Difficult cases (at least at my hospital) are reviewed in conference,
 I agree with you: your protocol (specially if it is based on DAKO'sprotocol) 
 should remain as is.
 The diagnosis differences should not determine a change.
 René J.

   *From:* Mark Tarango marktara...@gmail.com
 *To:* Wilson A wilson6...@yahoo.com
 *Cc:* histonet@lists.utsouthwestern.edu 
 histonet@lists.utsouthwestern.edu
 *Sent:* Tuesday, February 5, 2013 2:24 PM
 *Subject:* Re: [Histonet] HER-2

 I'd be interested to know if all your pathologists agree that the 2+ cases
 are 2+.  Is it possible that one of the pathologists is calling more cases
 as 2+ than the rest?  I would have a lot of questions before modifying the
 staining protocol.  It would be helpful you posted more info.

 Mark T.

 On Fri, Feb 1, 2013 at 6:52 PM, Wilson A wilson6...@yahoo.com wrote:

 
 Hi,
   Our pathologists are concerned we may be reporting too many 2+
  HER2’s.  Can someone  help with this?
 
   Thanks,
 Wilson
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Re: [Histonet] Re: Question

[Mark Tarango]

If anyone has any documentation that says the staining of IHC slides is NOT
high complexity it would help a histonetter out there.  I got an e-mail
from someone who is HT(ASCP)QIHC but does not have an AA degree.  Their lab
director is threatening their job saying they aren't qualified to do IHC
staining.  If anyone has something to refer to it would be helpful for this
person.  I already suggested contacting CAP and getting a written response.

I believe IHC is high complexity but not the staining portion.  Since no
result is being produced by the IHC tech how can this be high complexity?

thanks

Mark

On Wed, Jan 23, 2013 at 11:44 AM, Tim Higgins thiggin...@msn.com wrote:


 The professional interpretation is considered a high complexity test but
 not the actual technical component.



 Thanks,



 Timothy N. Higgins, HT (ASCP), QIHC

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Re: [Histonet] Question

[Mark Tarango]

The staining portion is not high complexity.  The reading of the slide is.

On Tue, Jan 22, 2013 at 10:04 AM, Courtney Pierce 
courtney.pie...@quintiles.com wrote:

 Are IHC high complexity test.

 Courtney Pierce
 IHC Specialist
 Quintiles
 Translational RD - Oncology
 Innovation
 Navigating the new health

 777 Oakmont Lane Suite 100
 Westmont,IL 60559

 Office: + 630-203-6234
 courtney.pie...@quintiles.com

 clinical | commercial | consulting | capital


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Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Mark Tarango]

It can't be used to just pull blocks.  The slides have to be reviewed and
the best block chosen by a pathologist.  If there is only one block then
the pathologist needs to look at the slides and determine if there is
enough tissue for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and
the best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out
over 30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy 
nagy_nata...@holyokehealth.com wrote:

 Hi everyone,
I just have a question about CPT code 88363, first can
 it be used for pulling blocks for Oncotype DX testing, also is there a time
 limit on when this code can be used? Does it have to be within a year, a
 month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


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Re: [Histonet] MDM2 Probe/Ventana

[Mark Tarango]

Hi Vicki,

I don't have a protocol but it should be very similar to the HER2 dual ISH.
 I'm assuming that you'll be using a probe targeted to chromosome 12 too
and not just doing the MDM2 (but maybe I'm wrong).  I would start there.

I'm working on validating MDM2 FISH and it's nearly identical to our HER2
FISH protocol, right down to denaturation and hybridization times.

Mark

On Wed, Dec 26, 2012 at 6:05 AM, Gauch, Vicki gau...@mail.amc.edu wrote:

 Hi,
 Does anyone have a protocol for the MDM2 Probe from Ventana that they
 could share ?  We have been unable to locate one thus far and our IHC
 Technical Specialist needs one.
 Thank you in advance for your assistance,

 Vicki Gauch
 AMCH
 Albany, NY




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[Histonet] moleculer testing

[Mark Tarango]

Hi Cynthia,

We do PCR for KRAS and BRAF.  You cant do these by FISH anyway, the
mutations are too small.

We do FISH for ALK and HER2.  For ALK,  there is no antibody that can
detect all the positive cases.  ALK FISH is the only method that can detect
ALK break aparts regardless of the fusion partner for ALK.  If you want to
do PCR for ALK, you will most likely be testing mRNA which isnt too stable
and since all the ALK mutations are not known, you will miss some.  This
could require more tissue too.

For HER2 there are some chromogenic ISH methods.  I still like FISH since
you can look at each color chanel individually.  I never worry that one
color is hiding the other and you can see each signal (no estimation of #
of signals in a cluster).

I dont think having a permanent record on a slide is the most important
thing for molecular testing.

Mark

On Monday, December 10, 2012, Cynthia Pyse wrote:

 Happy Monday Histonetter

 Who out there in Histoland is doing the testing for BRAF, KRAS, ALK, and
 HER2neu and what testing methods are you using. I prefer not to use FISH.
 Would rather have a permanent record of the slides.  Any information is
 welcome. Have a great week.

 Cindy



 Cindy Pyse, CLT, HT (ASCP)

 Laboratory Manager

 X-Cell Laboratories

 20 Northpointe Parkway Suite 100

 Amherst, NY 14228

 716-250-9235 etx. 232

 e-mail cp...@x-celllab.com



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Re: [Histonet] RE: recommended ICC protocol for use of Conjugated mouse antibodies with unconjugated mouse antibodies

[Mark Tarango]

Francie,

Conjugated antibodes CAN be detected by secondary antibodies.  So the
conjugation doesn't really hide the primary antibody to the secondary.
I've done it and seen it with biotin and FITC labeled antibodies.
Something to consider..

Mark

On Wed, Nov 7, 2012 at 11:43 AM, Frances Elizabeth Barron 
fbar...@stanford.edu wrote:

 Hi All,

 Hope this day finds you well.

 I have two antibodies in the lab that I would like to use together for
 ICC. One is a mouse anti-human pre-conjugated antibody, the other is a
 mouse anti-human unconjugated antibody. My question ishow to get these
 guys to play nice with one another or should I not be worrying about it?

 My thought is that I have to do this sequentially, (stain first with the
 unconjugated, stain with the secondary, then stain with the pre-conjugated
 antibody?). However, I'm questioning whether or not this is necessary.
 Would the mouse secondary react to a conjugated antibody? Does the process
 of conjugating hide the secondary epitope?

 I have had bad luck with loosing substantial signal from the first round
 of primary/secondary antibody staining in a sequential staining, so I'm
 loathe to do it (appreciate any advice on that issue if you have it!). I
 can find one of the antibodies raised in another animal, but it isn't
 validated in human or ICC (just predicted to work). Just trying to see if I
 can use what I have before I go out and buy another antibody that may or
 may not work.

 Appreciate any help with this!

 Best,
 ~Francie

 ***

 Francie Barron, Ph.D.
 Postdoctoral Fellow, Joseph Wu Lab

 Stanford University School of Medicine
 Lorry I. Lokey Stem Cell Research Building
 265 Campus Drive, Room G1105
 Stanford, CA 94305-5454

 Phone: (650) 724-5564 or (650) 724-9240
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Re: [Histonet] RNA isolation form stained slides

[Mark Tarango]

Have you tried using more sections during extraction?  Can you extract into
a smaller volume?

On Friday, November 2, 2012, Vanessa Orsini wrote:







 Hello,

 I need to extract RNA for a RT-PCR after Laser Micro
 Dissection on xGal stained slides.

 I tried using sections from unfixed frozen organs. I fixed the
 sections in EtOH70% for 10min and then I stained them with xGal for 3h at
 37°C.
 After air drying I cut out with the LCM and extract RNA with the PicoPure
 kit
 from Applied Biosystem. So far I didn’t manage to get enough RNA.

 I tried to add RNase inhibitors to all the solutions but it
 didn’t help.



 Any idea/suggestion?

 Do someone think it would be better to do a LacZ antibody staining
 on FFPE sections and extract RNA with an appropriate kit? The RNase would
 they
 be less active?



 Thank in advance for any help you can give me J Vanessa


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Re: [Histonet] ALK Positive lung slides

[Mark Tarango]

Hi Jill,

Have you tried using cases that you've already sent out to other labs and
they've reported as ALK-positive?  That is how we validated this assay in
my lab.  Most cases aren't very positive (you only have to find 15 cells
out of a hundred that you count to call the case positive if a second
observer agrees).  You're supposed to look around for positive cells using
your dual green/red filter.  Since most tumor cells show a fused yellow or
adjacent red  green signals, you really do have to look around for them a
little.  Because of that, I'd suggest doing the validation with cases that
have a range of % of positve cells.  If another lab calls a case positive
at 25% positive cells then you'd want to get that result too most of the
time.

That being said, I can send you some de-identified slides from a case of
metastatic (to a node) NSCLC that is super positive.  I do think it is
helpful to see what a case with a very high % of positive cells look
like.  This is what we're using as our positive control.  We think it's
better to have a tissue control than the cell cultured cells, processed
into cell blocks, that are on the slides sold by Abbott.  Send me your
address and fedex # if interested.

Mark


On Fri, Nov 2, 2012 at 9:20 AM, Johns, Jill jjoh...@cpallab.com wrote:

 Does anybody have any lung tissue slides that are known to be positive for
 the ALK gene rearrangement or know of a source to obtain them? We are
 trying to validate this FISH assay are having a very difficult time finding
 any positive specimens (we have Abbott's positive control slides, but they
 do not look like real tissue specimens)Thanks!!

 Jill A. Johns, MT(ASCP)SH, QCym, CCy
 Manager of Flow Cytometry and Molecular Diagnostics
 Central PA Alliance Laboratory (CPAL)
 1803 Mt. Rose Ave., Suite C3/C4
 York,  PA17403
 phone: (717) 851-4320
 fax:  (717) 851-1450
 email:  jjoh...@cpallab.commailto:jjoh...@cpallab.com


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Re: [Histonet] RNA isolation form stained slides

[Mark Tarango]

 I would try getting more sections from LCM and extracting into 7.5 uL, if
that volume will be enough to for your PCR reactions.

On Friday, November 2, 2012, Vanessa Orsini wrote:

 With the kit I'm extracting in 10ulthe problem is that I have too use
 few stained cells isolated with the LCM...so even if I increase the number
 of slides I'll never have a lot of material...

 Inviato da iPhone

 Il giorno 2 nov. 2012, alle ore 16:44, Mark Tarango 
 marktara...@gmail.com javascript:_e({}, 'cvml',
 'marktara...@gmail.com'); ha scritto:

 Have you tried using more sections during extraction?  Can you extract
 into a smaller volume?

 On Friday, November 2, 2012, Vanessa Orsini wrote:







 Hello,

 I need to extract RNA for a RT-PCR after Laser Micro
 Dissection on xGal stained slides.

 I tried using sections from unfixed frozen organs. I fixed the
 sections in EtOH70% for 10min and then I stained them with xGal for 3h at
 37°C.
 After air drying I cut out with the LCM and extract RNA with the PicoPure
 kit
 from Applied Biosystem. So far I didn’t manage to get enough RNA.

 I tried to add RNase inhibitors to all the solutions but it
 didn’t help.



 Any idea/suggestion?

 Do someone think it would be better to do a LacZ antibody staining
 on FFPE sections and extract RNA with an appropriate kit? The RNase would
 they
 be less active?



 Thank in advance for any help you can give me J Vanessa


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Re: [Histonet] Cassette Labeler

[Mark Tarango]

Hi Norm,

It sounds like he had something to contribute.  I don't think It shouldn't
matter where he works.  Someone else asked the question.

Mark
On Mon, Oct 29, 2012 at 10:00 AM, Norm Burnham norm.burn...@propath.comwrote:

 I didn't know that the Histonet site is being used as a medium for vendors
 to
 advertise their wares?  Your thoughts?
 Norm Burnham

 __
 Norm Burnham, MBA, MT(ASCP)
 Director, Anatomic Laboratory Operations
 ProPath - The Leader in Pathology Services
 1355 River Bend Drive
 Dallas, TX 75247
 norm.burn...@propath.com
 214.237.1602 Office
 214.237.1802 Fax
 214.709.7127 Cell

 To learn more about ProPath, please visit www.propath.com



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dustin
 Paul
 Campbell
 Sent: Monday, October 29, 2012 11:54 AM
 To: Kaye Ryan; Shelly Coker; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Cassette Labeler

 To All,

 TBS offers a compatible cassette and slide labeler that interfaces with
 APEASY. We have several accounts using both systems.  If addition
 information
 is needed please contact me directly. dcampb...@trianglebiomedical.com

 Hope this helps,




 Dustin Campbell
 Service Technician

 3014 Croasdaile Drive, Durham   NC  27705
  p 919.384.9393   f 919.384.9595
 dcampb...@trianglebiomedical.com

 Visit us online at www.trianglebiomedical.com and follow us on



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kaye Ryan
 Sent: Monday, October 29, 2012 9:42 AM
 To: Shelly Coker; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Cassette Labeler

 Please share that information with me also.

 Thanks,
 Kaye

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Shelly
 Coker
 Sent: Saturday, October 27, 2012 10:14 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Cassette Labeler

 Anyone out there with AP Easy software that has a cassette labeler
 interfaced
 with your LIS software that you would recommend?  We are looking at adding
 one sometime next year.  (PS...while responses from vendors are welcome, I
 really want to hear from someone working with the instrument...)

 Thanks!

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[Histonet] DAPI staining

[Mark Tarango]

Hi Carol,

Does this person put them in a freezer for 20 mins or so after DAPI
staining?  Someone once told me this helps with smearing.  I'm not really
sure if its true, but couldn't hurt to try it.

Mark

On Thursday, August 30, 2012, Barone, Carol wrote:

 Someone sent me a question regarding DAPI staining ( I generally  use
 Hoechst )...but anyway, they are having smearing of nuclear contents. I
 would guess that is from rupturing of the nuclear membraneover
 fixation? Over air drying? They are fixing 10 mins in cold acetone and 10
 mins to air dry. What do the experts out there say? This could be a
 learning moment for me , as well.
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Re: [Histonet] Formalin and Operating Rooms

[Mark Tarango]

Hi Debra,

Formaldehyde was listed as known to be a human carcinogen in the 12th
Report on Carcinogens (2011) put out by the Department of Health and Human
Services.  Here is a link
http://ntp.niehs.nih.gov/ntp/roc/twelfth/profiles/Formaldehyde.pdf

This is what is probably behind any recent changes to its use or
availability.

Johnson  Johnson just a few days ago announced that they're going to take
formaldehyde out of baby shampoo!  Yah!

Mark

On Mon, Aug 20, 2012 at 3:25 PM, Debra Siena dsi...@statlab.com wrote:

 Hi All,

 I would like to ask if anyone has heard of any new regulations or laws
 that state that the Operating Room can't have formalin available in the
 room so that they can place formalin onto the sample right away?  I was
 wondering if anyone has heard of this, if you could tell me more about
 where it is coming from so that I can access a copy of it.  We have had
 some inquiries and I have not heard of this.  I appreciate any help that
 you can give and sorry, that I don't have more information.  Best wishes.

   Debbie Siena, HT(ASCP)QIHC
 StatLab Medical Products
 Technical Support Manager
 407 Interchange Street | McKinney, TX 75071
 t: 800.442.3573 ext. 229 | f: 972.436.1369
 dsi...@statlab.com | www.statlab.com


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Re: [Histonet] Cyclin D1 IHC

[Mark Tarango]

Hi Donna,

We use Dako #M3635 at 1:100.  Mantle cell lymphoma is our control.

Mark
On Tue, Jul 24, 2012 at 8:16 AM, Suresch, Donna L.
donna_sure...@merck.comwrote:

 Hello Histonetters,
 Has anyone done IHC using Cyclin D1 antibody?  What vendor was used?  What
 control tissue was used?
 Thank you.
 Donna Suresch - Merck  Co.

 Donna L. Suresch
 Imaging Research Scientist
 Merck Research Laboratories
 Department of Imaging - West Point Campus
 Mail Stop:  WP44KOffice: WP44-H129
 770 Sumneytown Pike
 PO Box 4
 West Point, PA  19486-0004
 Phone:  215-652-7349
 Fax:  215-993-6803


 Notice:  This e-mail message, together with any attachments, contains
 information of Merck  Co., Inc. (One Merck Drive, Whitehouse Station,
 New Jersey, USA 08889), and/or its affiliates Direct contact information
 for affiliates is available at
 http://www.merck.com/contact/contacts.html) that may be confidential,
 proprietary copyrighted and/or legally privileged. It is intended solely
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Re: [Histonet] p53, control

[Mark Tarango]

Hi Cindy,

The one from Dako works well at a 1:200 dilution on the Ventana platforms
(catalog #M7001).  We use HIGH GRADE ovarian serous carcinoma as a positive
control.  You should get intense nuclear staining throughout the tumor with
this control.

Mark

On Thu, Jul 19, 2012 at 12:25 PM, Cindy Bulmer cjbul...@sbcglobal.netwrote:

 What type of tissue is everyone using for a positive control for this Ab,
 and are you using Mouse or Rabbit Ab?

 Thanks,
 Cindy

 Cynthia Bulmer HT(ASCP)QIHC

 IHC Supervisor, CTPL

 Waco, TX
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[Histonet] How to make HP control tissue?

[Mark Tarango]

Hi Willem,

H. pylori needs the low pH of the stomach to survive and grow.  It won't
naturally be in lung tissue.  When looking for HP, it's found on the mucosa
and gastric pits of the stomach.  Since you're looking for it in a specific
place, I think putting it in lung is a bad idea.

Your best bet for control tissue is a positive gastrectomy specimen.  You
could chop that up into a thousand pieces and never run out.  We've had
several cases like this.

Send me your address and a FedEx account number and I'll see about send you
a block or two.

Mark

On Wednesday, July 4, 2012, Hoekert, W.E.J. wrote:

 Hi Histonetters,

 Does anybody has experience in making your own HP control tissue? I am
 tired of using positive biopsies of patients since they are always almost
 finished.  I have heard of a procedure on making HP controls but I am not
 exactly sure how it is done.

 I have tried the following: I went to the microbiology department and
 asked for some freshly grown HP bacteria. We scraped them of the
 petridishes and put them in formalin. I had them fixed for 24 hours and
 then I injected them into some already fixed lung tissue and processed it
 as normal. But if I do a HP stain on that tissue, I don't see any bacteria.
 Probably they rinse off during the process.

 Can anyone tell me how to do it?

 Thanks in advance,

 Willem Hoekert
 OLVG
 The Netherlands


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Re: [Histonet] Billing 88342

[Mark Tarango]

I would think that if you're billing the client and not the insurance that
you could charge per block for the technical.  After all you're just
providing the stain to them.  In my opinion, the client should eat this
cost.  I would let the client know that you'd be billing this way before
staining the slides.  They could order 25 stains on a single part and you
would be in the red big time.  If you're billing the patient directly, you
would have to follow the rule of charging only once per specimen.

Mark

On Tue, Jul 3, 2012 at 9:43 AM, Victor A. Tobias vtob...@uw.edu wrote:

 Looking for other opinions from those who do consult/referral work.

 If a client sends in a request for a single antibody done on multiple
 blocks on a single specimen, do you bill the client for each tech component
 ? The client will do the interpretation.

 What happens in the above scenario if the request is to bill the patient?
 Knowing you get reimbursed for one, do you eat the other charges are make
 the client select the one block?

 We have run numbers on potential lost revenue and the number is
 significant.

 Victor


 Victor Tobias HT(ASCP)
 Clinical Applications Analyst
 Harborview Medical Center
 Dept of Pathology Room NJB244
 Seattle, WA 98104
 vtob...@u.washington.edumailto:vtob...@u.washington.edu
 206-744-2735
 206-744-8240 Fax
 =
 Privileged, confidential or patient identifiable information may be
 contained in this message. This information is meant only for the use of
 the intended recipients. If you are not the intended recipient, or if the
 message has been addressed to you in error, do not read, disclose,
 reproduce, distribute, disseminate or otherwise use this
 transmission. Instead, please notify the sender by reply e-mail, and then
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[Histonet] Billing IHC on MOHS

[Mark Tarango]

I'm trying to think this through.  Like Will said, if it can be shown that
you can charge immunos per block... I thought we could only charge IHC per
specimen these days.  Wouldn't each stage be a different specimen?  I would
think billing per block of the same stage would be over charging.  Why
would the logic change because this is MOHS?  It is still the same
pathology billing code after all (88342).  I would also think that if this
situation is coming up often that someone is having trouble reading the
HEs.

Mark

On Tuesday, June 19, 2012, William Chappell wrote:

 Well, I don't know if that settles that.

 I haven't responded, because I have not worked for a Mohs
 dermatopahtologist who runs Immunos (I have worked at numerous Mohs
 laboratories), however, this explanation is contradictory.  Each stain is
 reported only once per block, not per slide or per layer (stage). Yet the
 definition of a block, Tissue flattened by cutting into pieces, embedded,
 and frozen in mounting medium used by histotechnologists to embed tissue
 for frozen sections.  Every stage represent a new block in which slides
 are cut.  These two statements are contradictory and need clarification.

 Now, my own opinion (again I have talked with my dermatopathologist and
 billing specialist and they are as lost as we) is that by definition, Mohs
 is a frozen section diagnosis that must be made by the surgeon (i.e., for a
 Mohs to be a mohs the surgeon removing the tissue must diagnose the tissue
 -- look it up).  Every section taken, at every stage is a separate block of
 the same case.  In the event you can charge immunos per case, only one
 charge can be made.  If it can be shown that immunos can be charged per
 block (per the definition below), every immuno on every block from every
 stage can be charged.

 Now for the practicality -- we always start questions like this because
 medicare sets standards for billing that other insurance companies then
 adopt.  We should NEVER ask, what can we charge for, but should always
 ask, what work did we do that it is fair for a patient to pay for.
  Ignore what medicare and insurance companies say, bill clients for the
 work we perform and for the results they get.  How much more raw cost is
 there in staining two Mohs blocks with the same immuno?  Is it fair to
 charge a patient double the amount for MUCH less than twice the work?

 Will Chappell HTL(ASCP), QIHC

 On Jun 19, 2012, at 9:15 PM, Kim Donadio wrote:

  Great team work! Job well done and a absolute answer is given.
 
  Thank you
 
 
  
  From: Carol Torrence ctorre...@kmcpa.com
  To: 'Kim Donadio' one_angel_sec...@yahoo.com
  Cc: 'Weems, Joyce K.' joyce.we...@emoryhealthcare.org; 'Ingles
 Claire' cing...@uwhealth.org; histonet@lists.utsouthwestern.edu
  Sent: Tuesday, June 19, 2012 2:10 PM
  Subject: RE: [Histonet] Billing IHC on MOHS
 
 
  The following is the response I recived from a coding specialist at the
 American Academy of Dermatology.  I am trying not to be concerned that the
 reference is 6 years old but I think it clears up what we thought to be
 true.
  88342 for IHC
  88314 other “special stains”
  Here is the description for 88314 according to November 2006 cpt
 Assistant article, the companion piece to the AMA CPT Code Book.
  The work of processing and interpreting one routine stain is included in
 the procedure 17311- 17315. This stain is usually hematoxylin and eosin, or
 toluidine blue. If other special stains are necessary after one routine
 stain, then the code for special stains may be used (88314) as well as
 immunoperoxidase stains (88342) or decalcification procedures (88311).
 Special stains are not typically used and in most Mohs practices are of low
 frequency. Each stain is reported only once per block, not per slide or per
 layer (stage).
  AMA CPT definition of a Block:Tissue flattened by cutting into pieces,
 embedded, and frozen in mounting medium used by histotechnologists to embed
 tissue for frozen sections.
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Re: [Histonet] Billing IHC on MOHS

[Mark Tarango]

Carol,

Its nice to hear this isn't a regular thing.  In reading your original
question, it sounded like you were excited to be charging five times for
those immunos and you were ready to argue for it.  Apparently that was not
the case, you wanted more information and thoughts on the subject.

I once worked for a MOHS surgeon who had billing practices with which I did
not agree.  It was the reason I quit.  I'm glad there are people of high
integrity, such as your group, doing this work.

Mark

On Wednesday, June 20, 2012, Carol Torrence wrote:

 I have nothing more to add regarding this subject but would like to
 address concerns expressed here that touch on fairness, frequency, cost and
 the abilities of techs and surgeons.

 ** **

 This was a very rare incident involving scar tissue and tumor.  Our Mohs
 lab does not do immunos, our pathology lab does.  My quest was to learn
 more about what constitutes a block.  We stained 5 slides that were the
 same ‘stage’ (one block)….’to be sure there would be no ‘fall offs’. We do
 not want to put the patient through more waiting than necessary.   I was
 not trying to charge for each slide I stained or gouge the patient.  My
 quest was for “correct coding” not what “can” I charge.  I certified as a
 CPC in 2005 but do not practice in that field.  That said, I have a  good
 grasp as to the seriousness of the subject of coding and documentation.***
 *

 ** **

 I have 30 plus years of experience in histology including management.   I
 have always had the pleasure of working with physicians of high integrity
 and continue to do so in the area of dermatology.  The majority of my time
 has been spent in a large medical center where coding questions could be
 addressed ‘in house’ so to speak.  When I was consulted by the surgeon
 regarding coding this case, our search for the correct coding stems from
 the level of integrity we practice on a daily basis.  I consulted the 
 *American
 Society for Mohs Surgery* (*ASMS*) as was suggested and they do not offer
 coding advice.  I was told that they are an administrative office and
 suggested that I contact AAD.

 ** **

 Thanks for sharing your thoughts and listening to mine.

 ** **

 Have a good day!  Onward and upward!

 Carol M. Torrence, HT(ASCP)CM 

 ctorre...@kmcpa.com javascript:_e({}, 'cvml', 'ctorre...@kmcpa.com');***
 *

 ** **

 Confidentiality Note: This message is intended for use only by the
 individual or entity to which it is addressed and may contain information
 that is privileged, confidential, and exempt from disclosure under
 applicable law. If the reader of this message is not the intended recipient
 or the employee or agent responsible for delivering the message to the
 intended recipient, you are hereby notified that any dissemination,
 distribution or copying of this communication is strictly prohibited. If
 you have received this communication in error, please contact the sender
 immediately and destroy the material in its entirety, whether electronic or
 hard copy. Thank you

 ** **

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Re: [Histonet] Fluorescence-filters

[Mark Tarango]

Hi Gudrun,

i read about 15 ALK cases a week.  If you are seeing a lot of collagen
fibers around the tumor cells, I'd try increasing the digestion time of
your pepsin (especially if they were fixed for longer than usual).  Before
altering the pretreatment though, you would want to make sure that these
slides were not exposed to light for any period of time.  The discrete
signals of the probes can quickly fade (much faster than with IF stained
slides).  I'd also make sure the door is closed in your dark room.  If
there is light in the corner of your eye the signals can be hard to see.
 ALK FISH should be scored at high power under oil immersion (60-100x
objective).  If they are scoring at 40x, it could be a problem.

good luck!

Mark Tarango

On Tuesday, June 5, 2012, Eric Hoy wrote:

 We do a LOT of fluorescent microscopy in our immunology lab, so I have a
 bit
 of experience with fluorescence.

 The answer to your question depends on what type of filters you have in
 your
 microscope, and what type of light source is on the microscope.

 Older fluorescent systems used absorption filters, which were simply discs
 of coloured glass.  These filters had a fairly wide band-pass, so the
 fluorescence tended to be less than we see with interference filters.  The
 good news with these filters is that they are nearly indestructible (unless
 you drop and break them.)

 Interference filters are produced by vacuum deposition of a thin film of
 metal vapour on high-quality glass.  These filters usually have much
 sharper
 band pass characteristics than absorption filters.  They are also
 considerably more expensive.  If handled properly, these filters will last
 for decades, but improper cleaning and handling of the filters can shorten
 their lifespan.  I have also heard that prolonged exposure to solvent
 vapours (such as we find in a histology lab), can damage the filters,
 although I have not seen any filters that suffered this type of damage.

 I have seen interference filters that show delamination of the metal film
 over time.  In my experience these are older filters that were not produced
 with the current technologies, and filters that have been mishandled.
 Interference filters made in the past 20 years should last as long as the
 microscope, if they are properly handled.

 If you are seeing reduced fluorescence, I would suspect the light source as
 the most likely problem.  Halogen lamps have less intensity than mercury
 vapour lamps, which are less intense than metal halide lamps, which are
 less
 intense than LED sources.  We have converted all of our microscopes to LED
 sources.  If you are using an older HBO or halogen lamp, the age of the
 lamp, the initial wattage of the lamp, and the alignment can all affect the
 fluorescent output.

 As you identified, perhaps the most important aspect of immunofluorescence
 is the skill and experience of the person who reads the slides.

 Let me know if you have further questions.

 Eric Hoy

 ===
 Eric S. Hoy, Ph.D., SI(ASCP)
 Clinical Associate Professor
 Department of Medical Laboratory Sciences
 The University of Texas Southwestern Medical Center
 Dallas, Texas
 Email: eric@utsouthwestern.edu
 ===

 On 6/5/12 9:37 AM, Gudrun Lang gu.l...@gmx.at javascript:; wrote:

  Hi!
 
  Filters for fluorescencemicroscopy tend to burn out after a certain
  duration of usage. What duration?
 
  We have filters for FITC, TRITC, Dapi and a triplefilter. The
 working-hours
  are about 150 per year.
 
 
 
  What do you think? Is it time to change them.
 
  I have often bad feedback about weak signals, and I would not be
 surprised
  if the microscope is the culprit and not our protocol.
 
 
 
  Weak signals refer last times to ALK-FISH on lung biopsies. Well fixed
 but
  tumourcells mixed within collagenfibers.
 
  - and unfortunately unexperienced doctors on reading of this special
 probe.



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Re: [Histonet] Calponin

[Mark Tarango]

I think normal breast staining of myoepithial cells is the best control for
this antibody.   We use BioGenex's antibody (product # MU333-UC) at 1:200
on the Benchmark XT (CC1 mild, 8 mins primary antibody and ultraview DAB
detection).

Mark


On Fri, Mar 16, 2012 at 7:34 AM, Rene J Buesa rjbu...@yahoo.com wrote:

 DAKO monoclonal antibody at 1:1000 for 30 minutes, HIER citrate at pH6,
 colon cancer (+) case as control.
 René J.

 --- On Fri, 3/16/12, Chakib Boussahmain chak_...@yahoo.com wrote:


 From: Chakib Boussahmain chak_...@yahoo.com
 Subject: [Histonet] Calponin
 To: histonet@lists.utsouthwestern.edu
 Date: Friday, March 16, 2012, 9:14 AM


 Hello Histonet
 Does anyone use Calponin stain? if so can you share the protocol with us?
 What do you use for epitope retrieval?
 Many thanks in advance.
 Chakib Boussahmain
 Histotechnologist
 HTL(ASCP)
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Re: [Histonet] Humidity in Histology Lab

[Mark Tarango]

Is the humitity causing any problems or is the number just not what you're
used to seeing when you look at the readout?


On Tue, Aug 30, 2011 at 10:08 AM, Jill Cox jco...@yahoo.com wrote:

 Hi Histonetters!
 I am having humidity issues in a new lab in Long Beach Ca. It's 68%
 humidity inside lab. I have my own a/c system and have tried all settings
 including the dry setting. I had level down to 52% then it went back up. Is
 there something I can do to control this? I'm new to this area so don't know
 if humidity is year round or seasonal. What are you all using or doing if
 you have this problem? Thank you in advance, Jill

 Jill Cox, HT ASCP
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Re: [Histonet] Kappa/Lambda ISH background

[Mark Tarango]

How are you doing your ISH?  A commercial manual kit, home-made kit,
instrument?



On Fri, Aug 5, 2011 at 8:31 AM, Johnson, Nacaela 
nacaela.john...@usoncology.com wrote:

 Would anyone be willing to share their protocols for the Kappa and
 Lambda ISH?  I cannot get rid of the eosinophilic background staining.


 Thanks,



 Nacaela Johnson, B.S. HTL (ASCP)CM

 Histotechnologist

 KCCC Pathology

 12000 110th St., Ste. 400

 Overland Park, KS 66210

 Office:  913-234-0576

 Fax:  913-433-7639

 Email:  nacaela.john...@usoncology.com


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Re: [Histonet] CD117 - FDA APPROVED PROTOCOL

[Mark Tarango]

We're sticking to our IVD CD117/c-kit rabbit poly from Dako.

Mark
On Fri, Jul 29, 2011 at 10:42 AM, Vickroy, Jim vickroy@mhsil.comwrote:

 We are finding that the FDA approved protocol from Ventana does not stain
 some definite gi stromal tumors that stain quite well with another vendor's
 clone and protocol.  My pathologists are trying to decide what to do with
 this.   I would appreciate in hearing what kind of experience other labs
 have had when validating the FDA approved protocol for CD117.

 James Vickroy BS, HT(ASCP)

 Surgical  and Autopsy Pathology Technical Supervisor
 Memorial Medical Center
 217-788-4046


 
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Re: [Histonet] cam 5.2

[Mark Tarango]

We get ours from Becton Dickinson, product #349205.

Mark

On Thu, Jul 28, 2011 at 9:23 AM, Cynthia Pyse cp...@x-celllab.com wrote:

 Hello Histonetters

 What company is everyone buying their Cam 5.2 antibody from? Thanks for the
 information.



 Cindy Pyse, CLT, HT (ASCP)

 Laboratory/Histology Supervisor

 X-Cell Laboratories

 e-mail cp...@x-celllab.com





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Re: [Histonet] cam 5.2

[Mark Tarango]

I should have mentioned...we use it at 1:20 dilution.

Mark

On Thu, Jul 28, 2011 at 9:27 AM, Mark Tarango marktara...@gmail.com wrote:

 We get ours from Becton Dickinson, product #349205.

 Mark

   On Thu, Jul 28, 2011 at 9:23 AM, Cynthia Pyse cp...@x-celllab.comwrote:

 Hello Histonetters

 What company is everyone buying their Cam 5.2 antibody from? Thanks for
 the
 information.



 Cindy Pyse, CLT, HT (ASCP)

 Laboratory/Histology Supervisor

 X-Cell Laboratories

 e-mail cp...@x-celllab.com





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Re: [Histonet] Working Ventana Probe Protocols (EBER)

[Mark Tarango]

Hi Linda,

I had simliar trouble before I bought and added the extra HybReady to the
protocol (there is one that comes with the detection kit, but you need two
for good staining).  It will clean up your stain and A LOT and remove those
blue splotches that show up when you don't use it.

Mark

On Mon, Jul 25, 2011 at 9:39 AM, Sebree Linda A lseb...@uwhealth.orgwrote:

 Hi Marc and Histonetters (especially those using XTs),


 I've been trying to work up VMS's new EBER probe with less than stellar
 results.

 I have some questions about the protocols you sent via Histonet back in
 early June.  My questions/protocols are in red.

 You state the following:

 Depar 16 min  We can only select depar without an incubation
 timedoes your instrument allow you to select a time?  We are doing
 this on an XT, is this an instrument difference?  What instrument are
 you running your ISH onUltra by chance?

 Enzyme Protease 2 for 4 min. We are using ISH Protease 2 / 12 mins. as
 that was our protocol with the old stuff.

 ISH (EBER) probe 4 min. Same

 Denature @ 85 degrees for 12 min. Same

 Hybe 1 hour What temperature are you hybridizing at?

 3 stringency washes for 8 minutes each At what temperature?

 Blue detection for 20 minutes Same

 Counterstain for 4 min We use Nuclear Fast Red for 8 mins.

 Marc, are you using HybReady?

 Anyone else having had some success with the new EBER probe, reagents
 and software, feel free to comment.  Of the 3 specimens I've run, known
 positive soft tissue, bm bx and cell block, the soft tissue and bm bx
 had some positivity with the EBER DNP and U6 DNP but the cell block was
 negative with both.  There was also lots of blue smearing and haze over
 the slides.  I'm afraid this optimization/validation could end up being
 very expensive even with our 3 control tissues picked up together on
 single slides.

 Thanks for everyone's help,

 Linda


 Linda A. Sebree
 University of Wisconsin Hospital  Clinics
 IHC/ISH Laboratory
 DB1-223 VAH
 600 Highland Ave.
 Madison, WI 53792
 (608)265-6596


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[Histonet] Antibody suggestions for dog and cat

[Mark Tarango]

Could anyone suggest suituble antibodies for the following markers in dog
and cat tissue:

Vimentin, Pankeratin, CD3 and CD20, CD18, MelanA, Factor VIII

Also could use a suggestion for C-kit in Dog tissue only.

thanks

Mark
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Re: [Histonet] Paraffin Wax Waste Disposal

[Mark Tarango]

That's funny that you got cited for that.  I was surprised to learn what our
safety officer setup at my lab for paraffin disposal.  A commercial company
takes our paraffin and makes a product with it that is mixed with cedar
sawdust and paraffin wax for starting fires in the BBQ, fireplace, or
campfires, etc.  I wonder if consumers would want to use this product
knowing where the paraffin came from.  The company tested the paraffin and
said it's pure enough to meet their standards.  Just thought it was an
interesting so I decided to share.

Mark

On Fri, Jul 22, 2011 at 1:22 PM, Joanne Clark jcl...@pcnm.com wrote:

 Hi All, we had our CAP inspection yesterday and were cited for disposing of
 our waste wax from the processors in regular waste.  In all my 20+ years of
 working in histology I have never disposed of the dirty wax in biohazard
 waste.  Especially now with the newer processors that have very little carry
 over.  I know this is probably state regulated by is anyone aware of a
 regulation or documentation that states what the amount of hazardous
 chemical in a substance must be before it is considered hazardous?  And if
 so, does anyone know of a way to measure the amount of xylene in waste
 paraffin?

 Thanks in advance.
 Joanne Clark, HT
 Histology Supervisor
 PCNM
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Re: [Histonet] Controls with patient specimen on same slide

[Mark Tarango]

We do put controls on each slide in a case.  Sometimes it's just one slide
that failed in a run.  A batch control wouldn't tell you which slide failed
if there are no internal controls in the patient tissue.  I personally
wouldn't feel comfortable doing IHC with batch controls.
Mark

On Wed, Jul 20, 2011 at 6:18 AM, Rathborne, Toni 
trathbo...@somerset-healthcare.com wrote:

 All of these responses are great. So here's a follow up question.
 Do you place a control tissue on EACH slide if you have multiple blocks for
 a case, or just on one of the slides?

 -Original Message-
 From: Richard Cartun [mailto:rcar...@harthosp.org]
 Sent: Tuesday, July 19, 2011 6:40 PM
 To: histonet@lists.utsouthwestern.edu; Rathborne, Toni
 Subject: Re: [Histonet] Controls with patient specimen on same slide

 We do not put our positive control tissue on the test slide; we run batch
 controls.  Many of the unstained slides (breast, GI, and prostate biopsies)
 that we use for IHC testing are cut in our Histology Laboratory as part of a
 part-type slide protocol.  For example, we cut 7 slides, 2 sections on each
 slide, for breast biopsies and stain #1, 4, and 7 with HE, and then use (if
 needed) #2, 3, 5, and 6 for IHC.  Therefore, it would be very difficult for
 us to place the positive control tissue on the same slide.  In addition, I
 receive a lot of consult cases from other hospitals where they send us
 unstained slides for testing.  Once again, it would be difficult to place
 the positive control tissue on the same slide and it would slow us down in
 terms of starting those slides once they arrive.  However, I think the main
 reason we don't pursue putting the positive control tissue on the same slide
 is the fact that it would consume an enormous amount of control tissue.

 Richard

 Richard W. Cartun, MS, PhD
 Director, Histology  Immunopathology
 Director, Biospecimen Collection Programs Assistant Director, Anatomic
 Pathology Hartford Hospital
 80 Seymour Street
 Hartford, CT  06102
 (860) 545-1596 Office
 (860) 545-2204 Fax


  Rathborne, Toni trathbo...@somerset-healthcare.com 7/19/2011
  3:27 PM 

 Hi,
 I'm interested in knowing how many of you are performing ihc with the
 control tissue and the patient tissue on the same slide. I have seen slides
 available which have designated areas for each tissue to be placed so there
 will not be any confusion. If you're doing it, have you encountered any
 problems? What benefits have you noticed since implementing this process?
 Are your pathologists in favor of this?  If you're not, why not?
 Thanks,
 Toni


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 prohibited and may be unlawful.  If you are not the addressee, please
 promptly delete this message and notify the sender of the delivery error
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Re: [Histonet] Controls with patient specimen on same slide

[Mark Tarango]

Hi Toni,

We put the control on the same slide and after it's stained and coverslipped
we draw a line to seperate the control and patient tissue with each side of
the line having either a C or P written on it.

Mark
On Tue, Jul 19, 2011 at 12:27 PM, Rathborne, Toni 
trathbo...@somerset-healthcare.com wrote:


 Hi,
 I'm interested in knowing how many of you are performing ihc with the
 control tissue and the patient tissue on the same slide. I have seen slides
 available which have designated areas for each tissue to be placed so there
 will not be any confusion. If you're doing it, have you encountered any
 problems? What benefits have you noticed since implementing this process?
 Are your pathologists in favor of this?  If you're not, why not?
 Thanks,
 Toni


 CONFIDENTIALITY NOTICE
 This message and any included attachments are from Somerset Medical Center
 and are intended only for the addressee.  The information contained in this
 message is confidential and may contain privileged, confidential,
 proprietary and/or trade secret information entitled to protection and/or
 exemption from disclosure under applicable law.  Unauthorized forwarding,
 printing, copying, distribution, or use of such information is strictly
 prohibited and may be unlawful.  If you are not the addressee, please
 promptly delete this message and notify the sender of the delivery error
 by e-mail or you may call Somerset Medical Center's computer Help Desk
 at 908-685-2200, ext. 4050.

 Be sure to visit Somerset Medical Center's Web site -
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Re: [Histonet] er pr validation

[Mark Tarango]

Hi Anita,

We ran 40 cases and sent the same blocks to Phenopath for them to
run for comparision.  There is a requistion for doing an ER/PR validation on
their website.

Mark

On Tue, Jul 19, 2011 at 1:43 PM, anita dudley azdud...@hotmail.com wrote:


 I know this has been talked about before here but how many slides are
 people doing for er pr validation,  and are you doing it with another hosp?
 thanks so much for your help.

 anita dudley
 providence hosp
 mobile al
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Re: [Histonet] TTF-1

[Mark Tarango]

My lab uses the SPT24 clone for TTF-1.  Let me know if you'd like any
protocol info.

Mark
On Thu, Jul 14, 2011 at 11:47 AM, Inman, Anna anna.in...@stmarygj.orgwrote:

 Is anyone using the SPT24 clone for TTF-1 using the Ultraview platform
 on the Benchmark XT?

 Our pathologists have read this clone is supposed to be more sensitive
 than Ventana's 8G7G3/1 clone

 Thank you

 Anna



 anna.in...@stmarygj.org

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Re: [Histonet] IHC for H. pylori

[Mark Tarango]

We run the H. pylori IHC stain on all stomach specimens to improve turn
around time; however, when an inflammatory background is not appreciated by
the pathologist the charge is removed before sign-out and we eat the cost of
producing the slide.
Mark

On Wed, Jul 13, 2011 at 11:37 AM, Richard Cartun rcar...@harthosp.orgwrote:

 We only run IHC for H. pylori when the appropriate inflammatory background
 is present or when the patient has tested positive in the past and we
 receive a follow-up gastric specimen where bugs are not identified on HE.

 Running H. pylori IHC on every gastric biopsy is uncalled for and borders
 on Fraud and Abuse.

 Richard

 Richard W. Cartun, MS, PhD
 Director, Histology  Immunopathology
 Director, Biospecimen Collection Programs
 Assistant Director, Anatomic Pathology
 Hartford Hospital
 80 Seymour Street
 Hartford, CT  06102
 (860) 545-1596 Office
 (860) 545-2204 Fax


  Joanne Clark jcl...@pcnm.com 7/13/2011 2:22 PM 
  Hi All, was wondering how everyone does their IHC for H. pylori.  Do you
 automatically run it on all biopsies of the stomach, or do you screen first
 with a giemsa and run IHC on the ones that are negative by giemsa?  Thanks
 for the input!

 Joanne Clark, HT
 Histology Supervisor
 Pathology Consultants of New Mexico
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Re: [Histonet] pax2

[Mark Tarango]

The only problem here is that the Cell Marque antibody is currently on
backorder until at least the end of August from what I've heard.

Mark

On Thu, Jul 7, 2011 at 12:36 PM, Angela Bitting akbitt...@geisinger.eduwrote:

 CC1 standard, 32 min incubation with heat disabled. I use Cellmarques
 predilute
  and UltraView DAB detection.

  Dorothy Glass techman...@yahoo.com 7/7/2011 3:28 PM 
  Does anyone have a working protocol for pax2 on the Ventana Ultra or XT?
  Can you please share on Histonet?

 Dorothy Glass
 Supervisor, SEPA LABS
 Brunswick,Ga.
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Re: [Histonet] VENTANA ULTRA ER,PR,HER2

[Mark Tarango]

I think Barbara was referring to the approval for the ER, PR and Her2
antibodies being for use with the Benchmark XT stainers and that they aren't
yet approved for use on the ULTRA stainers using newer formulated bulk
reagents.  It's funny because they both use the same detection kits and
antibodies.  The formulation for the bulk reagents is different and it is a
new instrument, so I think that's why they need another approval from the
FDA.   The antibodies were already approved for use with the Benchmark XT
stainers.

We aren't running Ventana's new Dual ISH Her2 probe kit but we will be
looking at it.  It does seem to have some perks from the lab's point of
view.  The instrument does the entire ISH procedure/stain.  No more having
to mix the reagents for the pretreatment as with Abbott's Pathvysion Kit,
repeated pipetting, 22 x 22 coverslips or rubber cement.  It's even worse if
you don't have a VP2000 pretreatment unit, then you're stuck doing FISH
pretreatment by hand with coplin jars in waterbaths.   You don't need a
fluorescent scope to see the slide with Ventana's new kit and it would be
easy for the pathologist to make sure the correct area is scored for Her2
amplification (as can be a problem with the pathologist circling the area of
interest on an HE and the tech matching it up to another unstained serial
section with Her2 FISH).  I know a lot of labs use the Pathvysion kit off
label and dilute the probe and change other things from the original FDA
approved protocol.  Then again, even deparaffinzing the slide using xylene
is off label, since it was approved using Hemo-D substitute.  I imagine it
would be harder to go off label with Ventana's software.   I guess you've
got to weigh the pros and cons as with everything.

Mark Tarango

On Sun, Jul 3, 2011 at 2:28 AM, Gudrun Lang gu.l...@gmx.at wrote:

 The Her2 CB11 clone of Ventana is FDA approved. In the package insert of
 the
 Her2 4B5 clone is stated, that the antibody was compared to CB11 and shows
 same or better performance.
 The CB11 clone was exchanged through 4B5.

 On the NordiQC website Ventana 4B5 and Dako Herceptest are called FDA
 approved.
 http://www.nordiqc.org/Run-30-B10/Assessment/assessment-B10-HER2.htm

 So in my opinion the company wouldn't exchange its antibody, if it wouldn't
 be approved now or soon.

 Gudrun Lang



 -Ursprüngliche Nachricht-
 Von: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Patsy
 Ruegg
 Gesendet: Samstag, 02. Juli 2011 18:09
 An: barbara.cr...@lpnt.net; histonet@lists.utsouthwestern.edu
 Betreff: RE: [Histonet] VENTANA ULTRA  ER,PR,HER2

 I thought Ventana and Dako both had FDA approved Her2, I know Dako has
 Hercept Test.

 Patsy Ruegg, HT(ASCP)QIHC
 IHCtech
 12635 Montview Blvd. Ste.215
 Aurora, CO 80045
 720-859-4060
 fax 720-859-4110
 www.ihctech.net
 www.ihcrg.org

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 barbara.cr...@lpnt.net
 Sent: Tuesday, June 28, 2011 8:54 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] VENTANA ULTRA  ER,PR,HER2

 We are investigating getting the Ventana Ultra.
 I discovered that the ER, PR,  HER2 are not yet FDA approved.

 If you are using the Ventana Ultra how are you doing the ER, PR,  HER2?
 Do you use the Benchmark XT?

 Is anyone using the INFORM HER2 Dual ISH DNA Probe Cocktail Assay?




 ANTOINETTE CRILL,
 E-mail:  barbara.cr...@lpnt.netmailto:barbara.cr...@lpnt.net

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Re: [Histonet] Her/2

[Mark Tarango]

Hi Marcia,

Our slides for the Her2 CAP survey stained just fine.  We're using a Ventana
XT to stain for Her2 by IHC.

Mark
On Wed, May 4, 2011 at 9:54 AM, Marcia Funk fu...@mercyhealth.com wrote:

 CAP validation Her/2 slides not staining as clear as before.  Is anyone
 else having any issues ? We use the Ventana system and our daily IHC slides
 are staining
 and controls are working well.  Is there a coating on these slides from CAP
 ?
 Thanks for any help ?
 Marcia Funk HT,QIHC



 Marcia Funk
 Histology Laboratory
 Mercy Medical Center North Iowa
 Mason City, IA, 50401
 641-428-7907
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Re: [Histonet] Pax-2 on FFPE human tissue

[Mark Tarango]

Hi Dana,

Invitrogen sells a rabbit polyclonal anti-pax-2 antibody that works well on
FFPE tissues (Cat # 180483).  We use a multi-tissue control block for a
control.  It has normal kidney to show the endothelial cells around
glomeruli and renal tubules staining, clear cell renal cell Ca to show tumor
staining and a small piece of tonsil to show that B-cells stain.

Hope I helped,

Mark

On Fri, Apr 29, 2011 at 11:04 AM, Settembre, Dana sette...@umdnj.eduwrote:

 Hi All,
 Having a tough time looking for a reliable vendor for Pax-2.
 What vendors are you using?
 I need to use this on formalin fixed paraffin embedded human tissue.
 What control do you use?
 Thank you.

 Dana Settembre
 Immunohistochemistry Lab
 University Hospital - UMDNJ
 Newark, NJ

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Re: [Histonet] RE: [IHCRG] Ventana EBER DNP

[Mark Tarango]

If anyone has the catalog numbers for the probes, that would help.  I
couldn't get this info from Ventana when I asked.  They said we have to ask
another lab, even for catalog numbers!

Mark

On Tue, Apr 19, 2011 at 7:59 AM, Sheila Fonner sfon...@labpath.com wrote:

 SAME HERE!!!  I would love some help with this.  We are going to start
 doing
 the EBER with Ventana and have no idea where to start.



 Thanks in advance for ANY suggestions or  help you could give.



 Sheila, HT (ASCP)

 KDL Pathology

 Knoxville, TN





 From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
 Tallardy, Betty S
 Sent: Tuesday, April 19, 2011 10:55 AM
 To: ih...@googlegroups.com
 Subject: [IHCRG] Ventana EBER DNP



 Hi Group,

 Is anyone stuck in the EBER transition with Ventanas'

  probe changeover? The new product is supplied in a small vial and of
 course
 there are no suggestions about how to use the product.

 If anyone has some ideas on how to proceed with this, I would really
 appreciate some suggestions.

 I hate to waste such a pricey product trying to get something workable.

 Thanks in advance.

 Betty Tallardy, HT(ASCP)QIHC

 Dept. of Pathology

 Presbyterian Hospital

 200 Hawthorne Lane

 Charlotte, NC 28204

 704-384-5490

 bstalla...@novanthealth.org

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Re: [Histonet] CAP checklist, question ANP.23041.

[Mark Tarango]

Well I wouldn't try and use a Ph.D. in religous studies to qualify for high
complexity testing...

On Wed, Mar 16, 2011 at 4:36 PM, Mark Turner mtur...@csilaboratories.comwrote:

 Regarding CAP checklist, question ANP.23041.  The operation of the imaging
 system is performed by high-complexity testing personnel.



 We have a question regarding the qualifications of the operators.  The
 operators of the Aperio system are simply scanning entire slides to make a
 record for us prior to returning slides to the client.  Our operators make
 no evaluation of the image other than whether the scan is adequate for
 recording purposes, and this is verified by our medical staff prior to
 release of the materials.  In your opinion, does this constitute
 high-complexity testing and require CLIA compliant qualifications for the
 operators?

 Mark Turner, Ph.D, HT(ASCP) QIHC

 IHC / Histology Manager

 CSI Laboratories



 770-817-0817 x 394

 678-205-4669  FAX

 mtur...@csilaboratories.com mailto:nmon...@csilaboratories.com

 csilaboratories.com 
 https://csi-srv-007/exchweb/bin/redir.asp?URL=http://www.csi-labs.com/

 11525 Park Woods Circle

 Alpharetta, GA 30005



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Re: [Histonet] VALIDATION of a new clone

[Mark Tarango]

Hi Jim,

If it's a new clone then a new valiation should be performed.  I know of
many instances where one clone stains something that another doesn't.

If this is regarding Ventana not selling certains clones, those clones are
still available from Cell Marque directly.

Mark

On Mon, Mar 14, 2011 at 11:16 AM, Vickroy, Jim vickroy@mhsil.comwrote:

 I am wondering what others  are doing when they validate a new clone of a
 antibody that you have used in the lab for quite sometime.  When we
 introduce a new antibody obviously we go through a pretty extensive
 validation process including sampling of around 20 - 30 specimens, some
 presumed positive cases and some presumed negative cases.   We also try to
 include in the positive cases some that are strongly positive and others
 that are weakly positive.

 Lately several of our regular antibodies are now no longer available in the
 same clone.  I am interested in how different labs would validate the new
 clone to be used in the lab.  It would seem to me that the validation
 process would not have to be as extensive since we have already established
 that the antibody works well in our laboratory.   I know if it was the same
 clone but a different lot we would just do a comparison of the old lot with
 the new lot, but what about a different clone?   I realize that the methods
 are going to be diverse but I would appreciate hearing from others.


 James Vickroy BS, HT(ASCP)

 Surgical  and Autopsy Pathology Technical Supervisor
 Memorial Medical Center
 217-788-4046


 
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Re: [Histonet] Her2 Gastric cases

[Mark Tarango]

We do use the Ventana antibody for gastric cases.  There is some difference
in reading the slide for the pathologist.  I'll send you an article on it in
a seperate e-mail (so I can attach it).

Mark

On Fri, Mar 11, 2011 at 8:54 AM, Coppin, Margaret copp...@aruplab.comwrote:

 Hello,



 I am hoping someone can shed some light on whether or not Her2 4B5
 (Ventana's antibody) is okay to run on gastric biopsies. We are getting
 some client requests for this and I wonder if I should re-direct them
 toward the Dako Hercep Test instead.



 Thank you.



 Margaret G. Coppin, HT(ASCP)

 Technical Supervisor--Immunohistochemistry



 ARUP Laboratories

 500 Chipeta Way

 Salt Lake City, UT 84108

 (801)583-2787 X3869

 copp...@aruplab.com








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Re: [Histonet] Thanks

[Mark Tarango]

So cruel!

On Fri, Feb 25, 2011 at 8:13 AM, sgoe...@mirnarx.com wrote:

 It's sunshine and 75 here in Texas...nanny nanny...I do love my state's
 weather!!  We had shorts on yesterday =)

 Sarah Goebel, BA, HT(ASCP)
 Histotechnologist
 Mirna Therapeutics
 2150 Woodward Street
 Suite 100
 Austin, Texas  78744
 (512)901-0900 ext. 6912


 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia
 Pyse
 Sent: Friday, February 25, 2011 9:38 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Thanks

 Happy Friday Histonetters

 I want to thank everyone for the information on IHC strainers. This is
 just
 what I needed. People who actually use the machines on a daily basis
 giving
 their opinions. What a great forum we have. Hope everyone has a great
 weekend. Currently we are receiving 6-10 inches of snow, hey, what do
 you
 except in Buffalo. Thanks again.

 Cindy



 Cindy Pyse, CLT, HT(ASCP)

 Laboratory/Histology Supervisor

 X-Cell Laboratories

 716-250-9235

 e-mail cp...@x-celllab.com





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Re: [Histonet] Pin4cocktail negative controls

[Mark Tarango]

Hi Milton,

We use Biocare's Universal Negative Control Serum in place of the antibodies
in the negative control staining protocol.  We use normal prostate tissue to
show lack of p504s staining (to make sure it isn't overstaining normal
glands).  The prostate cancer in the tissue control shows lack of p63+ and
CK34BE12+ basal cells.

I hope this helps.

Mark
On Sun, Feb 20, 2011 at 12:00 PM, Gomez, Milton milton.go...@nyumc.orgwrote:

 Hello Histonetters,

 What are you folks using as a negative control when running Pin4 cocktail
 antibody.

 Thanks in advance,
 MG
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Re: [Histonet] QIHC

[Mark Tarango]

Its true, that test is too easy if you ask me.  Dako handbook is all you'll
need.

Mark

On Thu, Feb 10, 2011 at 5:46 AM, Houston, Ronald 
ronald.hous...@nationwidechildrens.org wrote:

 Read and digest the Dako book and you will sail through the exam. For a
 specialist type qualification it really is pitiful.

 Ronnie

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Michael Bradley
 Sent: Wednesday, February 09, 2011 4:30 PM
 To: Histonet@lists.utsouthwestern.edu
 Subject: [Histonet] QIHC

 Hi all

 I am preparing to take the QIHC exam.  Does anyone have any suggestions on
 what I should study?  What areas does the exam concentrate on and what
 research material I can use.

 Thanks
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Re: [Histonet] Benchmark Problems

[Mark Tarango]

Hi Mike,

Did you wash the bottles and rinse them and then pump the water through the
instrument before adding reagents and purging all again?  We had a problem
like this about 2 weeks ago.  I think someone loaded the wrong bulk reagent
onto the instrument.  Cleaning it, purging with water and adding newly made
bulk reagents fixed the problem for us.

Mark

On Wed, Feb 9, 2011 at 8:52 AM, Dessoye, Michael J mjdess...@wvhcs.orgwrote:

 Hello,

 We started having a strange problem with our Ventana Benchmark XT.  Out of
 the blue, our slides stopped staining.  With most antibodies we get no
 reaction, some that were very strongly positive are now only very lightly
 staining.  They do appear to counterstain OK.  Following Ventana's
 recommendations we completely changed out all of our bulk fluids, purged the
 lines, changed antibodies and DAB kits.  The last shot is some kind of
 instrument issue that's not triggering an error.  Anyone ever see anything
 like this?

 Mike Dessoye
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Re: [Histonet] TTF-1/background staining

[Mark Tarango]

Hi Paula,

You mentioned the staining in a liver specimen using the cell marque
antibody against TTF-1.  The clone named 8G7G3/1 that cell marque sells is
known to stain liver cells and liver cancer.  Your pathologist might be
interested in knowing that.  Here is a reference you can give him:
http://www.nordiqc.org/Run-23-B5/Assessment/Assessment-TTF-1.htm   It also
lists another clone that doesn't cross react with liver.

Mark

On Thu, Feb 3, 2011 at 12:38 PM, Paula Lucas plu...@biopath.org wrote:

 How can I eliminate the background or as you say, non-specific staining?



 I'm no expert here, I admit, and so I'm asking for your help and
 suggestions.  I have searched the Histonet archives, and a lot of what I'm
 seeing deals with animal or rodent tissues, and a lot of it was confusing
 to
 me.



 To give you a little background info:

 We use the Lab Vision stainer and the TTF-1 antibody we use is from Cell
 Marque.  We were having issues with this marker from Lab Vision, so we
 switched to TTF-1 a while ago.  We use the UltraVision LP detection kit
 from
 Lab Vision. It's a polymer driven detection kit.



 The antibody is a ready to use, and we have the time set at 30 minutes.



 We were getting the background staining on a liver specimen last week, and
 we also had this problem today on a lung case.  My doctor is getting
 frustrated, and wants me to do something about this, and to repeat the
 TTF-1
 stain on the lung, so if someone can give me some suggestions to try, I'm
 ready to try them.



 Thanks in advance,

 Paula

 Lab Manager

 Bio-Path Medical Group

 Fountain Valley, CA

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Re: [Histonet] Re: Pax-2

[Mark Tarango]

Thanks for the response.  I heard Cell Marque's rabbit died and they aren't
shipping any more ab until they get a new supplier.  If anyone is using an
antibody from someone other than Cell Marque, I'd be interested.

Thanks
Mark
On Thu, Jan 6, 2011 at 8:40 AM, Dessoye, Michael J mjdess...@wvhcs.orgwrote:

 Pax-2 (rabbit poly) is available from Cell Marque but in our experience it
 also has some cytoplasmic background staining.

 Mike Dessoye

 ---
 Message: 21
 Date: Tue, 4 Jan 2011 16:06:56 -0800
 From: Mark Tarango marktara...@gmail.com
 Subject: [Histonet] Pax-2
 To: histonet@lists.utsouthwestern.edu
Histonet@lists.utsouthwestern.edu
 Message-ID:
aanlktimha5h26-8o6fx4wuk5oplobrog4nexmpsnt...@mail.gmail.com
 Content-Type: text/plain; charset=ISO-8859-1

 Hi Histonet,

 My lab needs to switch vendors on this antibody.  Where is everyone buying
 Pax-2?  I found on their website that Invitrogen sells an IVD rabbit poly
 Pax-2 antibody.  Has anyone tried this or others?  What about Epitomics RUO
 rabbit mono?  Do all the antibodies give that cytoplasmic background
 staining that I've seen?

 I'd appreciate any info at all.

 Thanks

 Mark Tarango

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[Histonet] Pax-2

[Mark Tarango]

Hi Histonet,

My lab needs to switch vendors on this antibody.  Where is everyone buying
Pax-2?  I found on their website that Invitrogen sells an IVD rabbit poly
Pax-2 antibody.  Has anyone tried this or others?  What about Epitomics RUO
rabbit mono?  Do all the antibodies give that cytoplasmic background
staining that I've seen?

I'd appreciate any info at all.

Thanks

Mark Tarango
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Re: [Histonet] H pylori control on ventana immuno stainer

[Mark Tarango]

Hi Gloria,

Are you still looking for some good HP controls?  I could send you a block
of one of our controls if you're intersted.  I have blocks from
several different specimens.  Just let me know your address and fedex
number.

Thanks,

Mark

On Sun, Nov 21, 2010 at 12:30 PM, Gloria Cole gloria.c...@usa.net wrote:

 Hi,

 I am setting up a new lab and I am using the Ventana immuno stainer, does
 anyone know of any good HP controls I can purchase for now that will work
 well with the Ventana?

 Thanks,

 Gloria
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Re: [Histonet] p53 cytology controls

[Mark Tarango]

Hi Justin,

Do you run the p53 on a cell block or a cytospin or something different?

Mark
On Mon, Nov 8, 2010 at 12:36 PM, Justin Peters 
jpet...@bostwicklaboratories.com wrote:

 We receive requests from our pathologists for p53 IHC on urine but I am
 unsure as to what controls to use.  We currently use a FFPE tissue
 control (I understand that it is not correct) but up until now this
 issue has not been addressed.  Can anyone help with good cytology
 controls?  Thanks.



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Re: [Histonet] kappa and lambda by CISH

[Mark Tarango]

Hi Melissa,

We use it on bone marrow biopsies all the time on the Ventana XT.  We made
sure to include BMBXs in the validation.  We do get strange staining from
time to time, so it's particularily important to look at the negative
control for these.

Mark Tarango

On Tue, Oct 19, 2010 at 7:49 AM, Kuhnla, Melissa
melissa.kuh...@chsli.orgwrote:

 Hello All,
 Anyone currently running Kappa and Lambda by CISH?  I am in the process
 of validating the Ventana products.  Due to the ASR status of these
 products the vendor is extremely 'tight lipped'.  I have the stains
 worked up and have completed a 25 case validation. Does anyone know if
 this test is not to be used on decaled specimens??
 Thank you


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Re: [Histonet] negative controls

[Mark Tarango]

Hi Vikki,

It's best to run it all together and run a negative control for each
detection kit.

Mark
On Fri, Oct 15, 2010 at 7:25 AM, Victoria Baker bakevicto...@gmail.comwrote:

 Hi
 I have a hypothetical question to those who run IHC on Ventana instruments.
 Are you running your negatives with your patient/test cases or on a
 separate
 run?  Also, if you are doing this and have to use a different detection kit
 how do you work the QA/QC portion of this for CAP requirements.

 Thanks

 Vikki
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Re: [Histonet] negative controls

[Mark Tarango]

Hi Sarah,

It's better to have the control on the same slide.  There are slides that
work and slides that don't.  You know which ones are good and which ones are
bad because you have that control on each slide.  It's not always a complete
run that fails.  Yes, you CAN have a single batch control (it's not against
the rules), but best practice is to have a control on each slide.

You don't process a control for each case, you use a similarly processed
piece of positive tissue placed on each slide (except the negative which
should just have the patient tissue).


Mark
On Fri, Oct 15, 2010 at 8:47 AM, sgoe...@xbiotech.com wrote:

 So for every HP you do, you process a control cassette with the patient
 tissue cassette?  That seems like alot?  How do you get that many
 control tissues on a daily basis?  What do you do with the remaining
 tissue in the control block?  If you throw them away everyday, I would
 be interested in some of them.  How do you know what IHC stains the
 pathologist is going to order to know what control tissue to fix and
 process at the exact same time?  We have always just had a bunch of
 blocks that you cut a control from?  I understand that there is
 variability with processing, age, etc. not trying to be dense just still
 don't understand... Most places I have ever worked have control blocks
 that they cut a fresh control from everyday, then stain with the patient
 tissue.  If there are 3 HP cases, from what I am understanding, you guys
 are saying you need 3 controls for slides that are on the same machine,
 with the same reagents, same antibody, and same times.  Why couldn't you
 just have one for all 3 cases?  Then the next day have a fresh ONE for
 that day, date them, and file them.  So if you needed to see the HP
 control for October 15th, you could go pull the control for that day...

 Sarah Goebel, B.A., HT (ASCP)
 Histotechnician


 XBiotech USA Inc.

 8201 East Riverside Dr. Bldg 4 Suite 100

 Austin, Texas  78744

 (512)386-5107




  Original Message 
 Subject: RE: [Histonet] negative controls
  From: Rene J Buesa rjbu...@yahoo.com
 Date: Fri, October 15, 2010 8:33 am
 To: Sebree Linda A lseb...@uwhealth.org, sgoe...@xbiotech.com
 Cc: Histo Net list server HistoNet@lists.utsouthwestern.edu

 Because each tissue block has its own characteristics regarding fixation
 and processing some of which can influence the reactivity. If you have a
 bank of negative controls, how can you be sure that any of those blocks
 have received exactly the same treatment and reacted in the same way to
 the test block?
 The same goes for any bank of positives, so that is why you should have
 a positive control section in the same slide as the test section.
 René J.

 --- On Fri, 10/15/10, sgoe...@xbiotech.com sgoe...@xbiotech.com wrote:


 From: sgoe...@xbiotech.com sgoe...@xbiotech.com
 Subject: RE: [Histonet] negative controls
 To: Sebree Linda A lseb...@uwhealth.org
 Cc: Histo Net list server HistoNet@lists.utsouthwestern.edu
 Date: Friday, October 15, 2010, 11:17 AM


   Why do you need a negative control for each block if you are runn=
 ing
   the  same  antibody  on each patient block?  Is it just for case by c
  ase  reference  so  the negative is filed with the patient slide?  Why
   co=  uldn't  you  have  a control slide bank that was dated so all
 the
   slides you d= id on that day, on that run, could be referenced back
 to
   that control? = ; Just curious?

   Sarah Goebel, B.A., HT (ASCP)

   Histotechnician= br

   XBiotech USA Inc.

   8201 East Riverside Dr. Bld= g 4 Suite 100

   Austin, Texas  78744

   =

   (512)386-= 5107

    Original Message 
   Subject: RE: [Histonet] negative controls
   From: Sebree Linda A [1]lseb...@= uwhealth.org
   Date: Fri, October 15, 2010 8:08 am
   To:  Victoria  Baker  [2]bakevict= o...@gmail.com, Histo Net
 list
   server
   [3]histo...@lists.uts= outhwestern.edu
   We run negative controls on every block of a case within the same
 run.
   On autopsy cases, we only run 1 negative per tissue type, within the
   same run...this is the only exception to the rule of 1 negative per
   block.
   Linda A. Sebree
   University of Wisconsin Hospital  Clinics
   IHC/ISH Laboratory
   DB1-223 VAH
   600 Highland Ave.
   Madison, WI 53792
   (608)265-6596
   -Original Message-
   From: [4]histonet= -boun...@lists.utsouthwestern.edu
   [[5]mailto:histon=  et-boun...@lists.utsouthwestern.edu]  On Behalf
 Of
   Victoria
   Baker
   Sent: Friday, October 15, 2010 9:26 AM
   To: Histo Net list server
   Subject: [Histonet] negative controls
   Hi
   I have a hypothetical question to those who run IHC on Ventana
   instruments.
   Are you running your negatives with your patient/test cases or on a
   separate
   run? Also, if you are doing this and have to use a different
 detection
   kit
   how do you work the QA/QC portion of this for CAP requirements.
   Thanks
   Vikki
   

Re: [Histonet] Problems with listserv

[Mark Tarango]

* *I think the chatter just died down since many of us are at the NSH
symposium.

Mark

On Wed, Sep 29, 2010 at 8:49 AM, Feher, Stephen sfe...@cmc-nh.org wrote:

 I am having some issues with receiving email from the listserv.  I was
 successfully receiving email and it suddenly stopped.  If I have been
 unsubscribed, please re subscribe my email address.

 sfe...@cmc-nh.org

 Thank you,

 Steve


 Stephen A. Feher, MS, SCT (ASCP)

 Pathology Supervisor

 Catholic Medical Center

 100 McGregor Street

 Manchester, NH 03102

 603-663-6707

 sfe...@cmc-nh.org mailto:sfe...@cmc-nh.org


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Re: [Histonet] Napsin A

[Mark Tarango]

Hi Jessica,

Our control for this antibody is normal lung, adenocarcinoma of the lung,
and thyroid.  We use this same block for other stains too (TTF-1,
sufactant-A, EMA, etc.) but if I were going to make a control block specific
for this antibody, I would want to include a squamous cell carcinoma of the
lung to show that it's negative for napsin-A.  You may see a very tiny
amount of staining on the squamous cell carcinomas where the
pulmonary-aveolar macrophages are eating up the keratin, but the tumor cells
will be negative.

Ovarian carcinomas can also stain.  I would include a few of those too so
the docs can see what that looks like.
I would also make sure to include thyroid and thyroid carcinomas.  With a
particular vendor's antibody, napsin-a seems to stain thyroid (I don't want
to name names), but with Novocastra's antibody it doesn't.  It was helpful
to our pathologists to have an antibody that doesn't stain thyroid.

Thanks
Mark

On Mon, Sep 13, 2010 at 8:57 AM, Jessica Piche jhist...@yahoo.com wrote:

 Hello Everyone,

 We are in the process of working up Napsin A. I was wondering what tissue
 others are using for multi tissue blocks to validate the anitbody? Thanks,


 Jessica Piche-Grocki, HT(ASCP)
 Waterbury Hospital, CT



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Re: [Histonet] FITC on Ventana Ultra

[Mark Tarango]

Hi Nita,

I agree that buffer should be used as the negative control reagent.  If you
have a seperate slide that is cut and put into buffer, just coverslip it
with the same mounting media and you have a negative control.  All the
instrument is doing is putting on the antibody and then rinsing it off.  So
your negative control is a slide that didn't get the antibody (just
buffer).  Makes sense to me.  If you get ventana to add a negative control
to their software it would do just the same thing as coverslipping from
buffer, but you'd be paying ventana for it.

Mark
On Fri, Aug 27, 2010 at 7:30 AM, Nita Searcy nsea...@swmail.sw.org wrote:

 Any users out there that have had A CAP inspector question negative control
 on the instrument? In regard to CAP question ANP.21850 in which the notes
 state, A negative reagent control in which the patient tissue is processed
 in an identical manner to the test specimen but with the primary antibody
 omitted must be performed for each patient test specimen?

 Below is the response from Ventana regarding negatives I am curios if CAP
 accepts these methods??

 In regards to the question below about running a negative control fitc,
 there are two ways to accomplish this.



 1) There is not currently a place in the protocol to select a fitc
 negative. We can still be compliant by ensuring the negative slide is
 treated the same as the patient. The only reagent the patient slide is
 exposed to, besides the fitc antibody, is reaction buffer. Running a
 negative can be accomplished by applying reaction buffer to the slide and
 letting it incubate for the same amount of time. Next, it is important to
 ensure both slides are coverslipped in the came mounting media.
 2) The second method is to utilize internal negatives that are already
 present within the patient tissue. However, this is difficult when running
 Fitc antibodies such as IgG, IgM, etc.
 We are looking into adding this addition into the software. Many customers,
 however have expressed that they would rather utilize the slide drawer for
 another patient slide, rather than running the fitc negative.

 Am interested in your comments.
 Thanks





 Nita Searcy, HT/HTL (ASCP)
 Scott and White Hospital
 Division Manager, Anatomic Pathology
 2401 S. 31st. Street
 254-724-2438
 Temple, Texas, 76502
 nsea...@swmail.sw.org


 254-724-2438


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[Histonet] Re: Anti-IDO (indolamine 2,3-dioxygenase)

[Mark Tarango]

Here I am answering my own question.  I just remembered where I used to get
this antibody.  It's from Millipore/Chemicon.

Mark
On Mon, Aug 9, 2010 at 7:53 AM, Mark Tarango marktara...@gmail.com wrote:

 Hi Histonet,

 Does anyone use an Anti-IDO on FFPE tissues?  Would you please let me know
 where you purchase it?

 Thank you,
 Mark Tarango

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Re: [Histonet] p63 from Ventana

[Mark Tarango]

I've been told by a Biocare Salesperson that the BC4A4 clone is the same
exact clone as 4A4.  The BC in front of 4A4 just means Biocare.  I don't
think that Ventana sells a concentrate of this antibody.  You'd want a
concentrate for your PIN4 so you're probably better off sticking with your
current antibody.

Mark
On Mon, Aug 9, 2010 at 11:46 AM, Adrienne Aperghis Kavanagh 
aaperg...@uspath.com wrote:

 Hi Everyone,

 We have been using the p63 (BC4A4) antibody from BioCare for the past 5
 years with great results.  We use it as a component on our triple stain for
 prostate.  I have been seeing ads on the Ventana website for their p63 and
 was wondering if anyone else had used their p63 and in what immuno?  Any
 feedback would be greatly appreciated!!!

 Thanks again!



 Adrienne Aperghis Kavanagh
 US PATH
 30 W. Century Road
 Suite 255
 Paramus NJ 07652


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Re: [Histonet] Using GEWF solution and IHC staining

[Mark Tarango]

Ethanol/alcohol is what will process the specimen.  If the tissue is fixed
would it really matter that the tissue came into contact with ethanol?

Mark
On Tue, Jul 27, 2010 at 11:03 AM, Hayes, Randi (HorizonNB) 
randi.ha...@horizonnb.ca wrote:

 At a recent conference, our PA learned of using GEWF (glacial acetic acid,
 ethanol, distilled water, 40% formaldehyde) solution as an aid for Lymph
 Node retrieval in Colorectal Cancer resections.  Although a good idea, I'm
 wondering how safe it is to use when staining for IHC.  Does anyone have
 much experience with this or know of a study (studies) that have been done
 to verify that the ethanol is not destroying antigen sites?  We're a little
 concerned..


 Randi Hayes, MLT
 Histology Supervisor / Superviseur d'Histologie
 Horizon Health Network / Réseau de santé Horizon
 (506) 860-2157
 randi.ha...@horizonnb.ca http://www.me.com/mail/
 www.HorizonNB.ca http://www.horizonnb.ca/ http://www.horizonnb.ca/



 /pre--- Horizon Health Network Disclaimer ---brbrThis e-mail
 communication (including any or all attachments) is intendedbronly for the
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Re: [Histonet] Biocare P63 with Dako Flex detection

[Mark Tarango]

Hi Cathy,

I'm using their p63, but on the Ventana XT with ultraview DAB detection.  We
use it at 1:200 and get a strong signal in both breast and prostate
tissues.

If you haven't tried staining various cases of prostate, I'd suggest it.
Sometimes I'll get a prostate that stains the basal cells nicely and other
times the signal is much weaker.  It might have something to do with
fixation in the resection specimens.  If you can't get a strong signal in a
prostate needle biopsy, then it's definately a staining issue and not the
tissue.

I got stuck trying to stain a bad prostate once and wouldn't want that to
happen to you.

Good Luck!

Mark

On Wed, Jul 21, 2010 at 12:46 PM, cathy.crump...@tuality.org wrote:


   Hi  all, I have ran into a troubleshooting dilemma and was wonderi ng
   if  anyone  else could help.  Is there anyone out there that uses the
 Biocare  Medical  P63 concentrated antibody with the Dako TRS and Flex
   detec  tion  kit?  I am curious what dilution you use and if you have
   to  do  an  ything  special  for  the  P63 (extra long retrieval, DAB
   enhancer, etc).  I am having problems getting a strong signal for the
   basal cells in prosta te.  I am running it at 1:50 with 40 minutes in
   pH  9  retrieval  (whichseems  too  long to me but does help).  A
   dilution with the same protoco l at 1:100 is way too light.

   Thanks,
 /DIV
   Cathy Crumpton HT(ASCP), Histology Lead
   Tuality Community Hosp ital
   Hillsboro, OR 97123
   (503)681-1292
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Re: [Histonet] H. pylori RTU antibody

[Mark Tarango]

Biocare has a rabbit polyclonal anti-HP ab and a mouse monoclonal.  Which
one do you use?

Mark
On Wed, Jun 30, 2010 at 8:05 AM, McMahon, Loralee A 
loralee_mcma...@urmc.rochester.edu wrote:

 Use the RTU from Biocare.  It is clean and easy.  You can use it with the
 Dako Flex Kit.

 Loralee McMahon, HTL (ASCP)
 Immunohistochemistry Supervisor
 Strong Memorial Hospital
 Department of Surgical Pathology
 (585) 275-7210
 
 From: histonet-boun...@lists.utsouthwestern.edu [
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Drew Meyer [
 41dm...@gmail.com]
 Sent: Wednesday, June 30, 2010 10:41 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] H. pylori RTU antibody

 I was just wondering if a lot of people use the RTU antibodies available
 out
 there for H. pylori.  Currently, we're testing the RTU from Dako and we're
 having issues with a lot of background staining.  After tweaking the
 protocol a lot, I've managed to get a lot of it removed, but there are
 still
 some issues.  I was wondering if most people are making their own dilutions
 to get a better stain or if the RTUs are commonplace.  I appreciate the
 feedback!

 Thanks,
 Drew
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Re: [Histonet] New CAP question ANP.22760

[Mark Tarango]

You'll have to use prep kit stickers and duplicate protocols to do it, but
it is possible.  You just need to copy the protocol and save it as another
number, then change the primary antibody to a prep kit sticker save again
and then put that sticker on the dispenser.  Then you need to print stickers
for both protocols and stick them on two controls slides and run.

I admit its a little bit of a pain.

Mark

On Fri, Jun 18, 2010 at 9:41 AM, Mike Pence mpe...@grhs.net wrote:


 I don't think I can do this with the automated system we are currently
 using. Ventana. Does any other Ventana users know if you can do this in
 parallel

 Mike
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ellen
 Yee
 Sent: Thursday, June 17, 2010 7:21 PM
 To: Laurie Colbert
  Cc: histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] New CAP question ANP.22760


 Sorry, I should have included it.

 ANP.22760  Are new lots of antibody and detection system reagents tested
 in parallel with old lots?  (NOTE: New lots of primary antibody and
 detection system reagents must be compared to the previous lot using an
 appropriate panel of control tissues.)

 Ellen Yee
  _

  From: Laurie Colbert [mailto:laurie.colb...@huntingtonhospital.com]
 To: Ellen Yee [mailto:e...@dpmginc.com]
 Sent: Thu, 17 Jun 2010 08:47:38 -0700
 Subject: RE: [Histonet] New CAP question ANP.22760

 Can you give us the wording of that question/checklist item? Laurie

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ellen
 Yee
 Sent: Wednesday, June 16, 2010 10:10 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] New CAP question ANP.22760

 How are IHC labs complying with this question? What is considered an
 appropriate panel of control tissues? What do you stain to test your
 detection systems?

 Ellen Yee

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Re: [Histonet] AlphaSMA staining

[Mark Tarango]

Hi Phebe,

I can't be sure about this since you didn't post your protocol, but
alpha-SMA is an antibody that does not require antigen retrieval.  If you're
doing some kind of retrieval, I'd suggest trying it without.

Thanks

Mark

On Wed, May 26, 2010 at 6:56 PM, Phebe Verbrugghe vphe...@yahoo.com wrote:










 Hello everyone,

 We are trying to stain alpha smooth muscle actin (on mouse/human liver)
 using the A5228 Sigma antibody and are getting nuclear staining while this
 stain should be cytoplasmic. Does anyone have experience with nuclear
 staining of alphaSMA? Is there anything I can do to avoid nuclear staining
 or can anyone recommend me another good anti- alphaSMA antibody that works
 on both mouse and human formalin fixed paraffin sections?

 Thank you very much in advance!

 Phebe




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Re: [Histonet] weekend fixation

[Mark Tarango]

I just realized the question came from Belgium.  I have no idea how they do
things there.

Mark

On Thu, May 20, 2010 at 2:14 PM, Mark Tarango marktara...@gmail.com wrote:

 Yes, you can go up to 72 hours for ER/PR (new CAP/ASCO guidlines), but if
 you do HER2 the maximum is still 48 hours.  I'm assuming you want HER2 as
 well, so your best option would probably be to hold the tissues in 70%
 alcochol on the processer until Sunday night.

 Mark Tarango
   On Thu, May 20, 2010 at 12:54 PM, Liz Chlipala l...@premierlab.comwrote:

 I was talking to Peggy Wenk over the weekend at the MSH meeting and they
 had a paper that was published regarding fixation and ER/PR staining
 sensitivity etc.  The biggest problem that they reported is
 underfixation is much worse than over fixation.  I think a minimum of 10
 hours of fixation demonstrated good results and that intensity of
 staining started to decrease but not by much at 48 hours.  I would be
 more concerned over underfixation than overfixation.  Also the new ER/PR
 guidelines state its acceptable to have samples in fixative for 72
 hours.  Maybe Peggy can post the link to this paper.

 Liz

 Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
 Manager
 Premier Laboratory, LLC
 PO Box 18592
 Boulder, Colorado 80308
 office (303) 682-3949
 fax (303) 682-9060
 www.premierlab.com


 Ship to Address:
 1567 Skyway Drive, Unit E
 Longmont, Colorado 80504

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of LINDA
 MARGRAF
 Sent: Thursday, May 20, 2010 1:04 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] weekend fixation


   Histonetters:
 Here's a message I was asked to post..

 Dear Colleagues,
  I  have the following question concerning tissue processing. We do a
 lot of IHC work on NF fixed tissue. To standardize and minimize the
 effect of NF fixation, we fixate the tissue always for 24h. This is of
 course a problem for tissues taken on Friday. In the past, we asked our
 technicians to come on Saturday to embed the tissues in paraffin.
 Unfortunately, this is not possible anymore, and that is why I need your
 advice. What would you suggest ? 1) to leave the tissue in NF until
 Sunday evening and start processing, or 2) to keep the fixation time (24
 hours) and leave the tissue in alcohol 70% until Sunday evening and then
 start processing.
  Thanks for your advice.
 Kind regards,
 Wim.


 Prof. dr. Wim Van den Broeck, DVM, MSc, PhD
 Cell biology and Histology
 Department of Morphology - Faculty of Veterinary Medicine
 Ghent University
 Salisburylaan 133, B-9820 Merelbeke, BELGIUM



 Please consider the environment before printing this e-mail.

 This e-mail, facsimile, or letter and any files or attachments
 transmitted with it contains
 information that is confidential and privileged. This information is
 intended only for the
 use of the individual(s) and entity(ies) to whom it is addressed. If you
 are the intended
 recipient, further disclosures are prohibited without proper
 authorization. If you are not
 the intended recipient, any disclosure, copying, printing, or use of
 this information is
 strictly prohibited and possibly a violation of federal or state law and
 regulations. If you
 have received this information in error, please notify Children's
 Medical Center Dallas
 immediately at 214-456- or via e-mail at priv...@childrens.com.
 Children's Medical
 Center Dallas and its affiliates hereby claim all applicable privileges
 related to this
 information.


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Re: [Histonet] weekend fixation

[Mark Tarango]

Yes, you can go up to 72 hours for ER/PR (new CAP/ASCO guidlines), but if
you do HER2 the maximum is still 48 hours.  I'm assuming you want HER2 as
well, so your best option would probably be to hold the tissues in 70%
alcochol on the processer until Sunday night.

Mark Tarango
On Thu, May 20, 2010 at 12:54 PM, Liz Chlipala l...@premierlab.com wrote:

 I was talking to Peggy Wenk over the weekend at the MSH meeting and they
 had a paper that was published regarding fixation and ER/PR staining
 sensitivity etc.  The biggest problem that they reported is
 underfixation is much worse than over fixation.  I think a minimum of 10
 hours of fixation demonstrated good results and that intensity of
 staining started to decrease but not by much at 48 hours.  I would be
 more concerned over underfixation than overfixation.  Also the new ER/PR
 guidelines state its acceptable to have samples in fixative for 72
 hours.  Maybe Peggy can post the link to this paper.

 Liz

 Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
 Manager
 Premier Laboratory, LLC
 PO Box 18592
 Boulder, Colorado 80308
 office (303) 682-3949
 fax (303) 682-9060
 www.premierlab.com


 Ship to Address:
 1567 Skyway Drive, Unit E
 Longmont, Colorado 80504

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of LINDA
 MARGRAF
 Sent: Thursday, May 20, 2010 1:04 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] weekend fixation


   Histonetters:
 Here's a message I was asked to post..

 Dear Colleagues,
  I  have the following question concerning tissue processing. We do a
 lot of IHC work on NF fixed tissue. To standardize and minimize the
 effect of NF fixation, we fixate the tissue always for 24h. This is of
 course a problem for tissues taken on Friday. In the past, we asked our
 technicians to come on Saturday to embed the tissues in paraffin.
 Unfortunately, this is not possible anymore, and that is why I need your
 advice. What would you suggest ? 1) to leave the tissue in NF until
 Sunday evening and start processing, or 2) to keep the fixation time (24
 hours) and leave the tissue in alcohol 70% until Sunday evening and then
 start processing.
  Thanks for your advice.
 Kind regards,
 Wim.


 Prof. dr. Wim Van den Broeck, DVM, MSc, PhD
 Cell biology and Histology
 Department of Morphology - Faculty of Veterinary Medicine
 Ghent University
 Salisburylaan 133, B-9820 Merelbeke, BELGIUM



 Please consider the environment before printing this e-mail.

 This e-mail, facsimile, or letter and any files or attachments
 transmitted with it contains
 information that is confidential and privileged. This information is
 intended only for the
 use of the individual(s) and entity(ies) to whom it is addressed. If you
 are the intended
 recipient, further disclosures are prohibited without proper
 authorization. If you are not
 the intended recipient, any disclosure, copying, printing, or use of
 this information is
 strictly prohibited and possibly a violation of federal or state law and
 regulations. If you
 have received this information in error, please notify Children's
 Medical Center Dallas
 immediately at 214-456- or via e-mail at priv...@childrens.com.
 Children's Medical
 Center Dallas and its affiliates hereby claim all applicable privileges
 related to this
 information.


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