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 time is greater than 1 hour or if the 

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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