Re: [Histonet] Detection Systems

[Sally Price via Histonet]

Like Amos, I have extensive experience in working with biotin/streptavidin
(BSA) detection systems, and use them primarily because, in my analyses,
such reagents cost about half as much as polymers kits in the same volume
containers.  In addition, knowing that endogenous biotin can be problematic
in a small number of different tissues, our (clinical) lab simply avoids
using BSA in those situations.  As required by CAP standards, our IHC
policy acknowledges that we’ve conducted the necessary testing to determine
if/when endogenous biotin makes interpretation challenging, and refrain
from doing so when applicable.

Finally, although I don’t agree with Carl’s belief that one MUST use the
DAB chromogen+substrate components offered by the same vendor as the
detection components, we DO purchase such a reagents from the same
manufacturer simply because they’re more inclined to assist us in
troubleshooting if we run into a problem with EITHER reagent.
Sally


On Sun, Feb 6, 2022 at 2:40 PM Hobbs, Carl via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> As I stated:
> Polymer systems only work GREAT when you also use the DAB in the kit
> You pay dearly for it
> Check out my comparison in Histonet Images
> Carl
> Carl Hobbs FIBMS
> Histology and Imaging Manager
> Wolfson CARD
> Guys Campus, London Bridge
> Kings College London
> London
> SE1 1UL
>
>
>
> 020 7848 6810
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
-- 
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Re: [Histonet] Antibody diluent

[Sally Price via Histonet]

Maria:
Personally, I believe in using the right tool for the job, and have learned
a lot about the science of IHC from Biocare.  I recently received an email
from them with a link to download a paper from their website that’s all
about diluents and I found this information very useful.  I would encourage
others to review this information if they want to learn why using a
universal reagent best option.
Sally--
Sally Price
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Re: [Histonet] ADH5 multiplex procedure

[Sally Price via Histonet]

-- Forwarded message --
From: "Kurth Virginia L." 
To: "histonet@lists.utsouthwestern.edu" 

Virginia:
I’m pretty certain that the ADH5 multiplex procedure is performed at CSI
Labs in the Atlanta area.  And I don’t think that biocare refers to it by
that name any more either - I think they just spell out all the antigens
that the antibodies in this cocktail can identify, like CK-5/14+p63+CK-7/18.

Sally

- - - - - - - - - - -

Date: Tue, 15 Oct 2019 14:48:40

Subject: [Histonet] ADH5
Does anyone know if there is a reference lab that performs ADH5.
(particular the BioCare multiplex stain?)

Thanks Ginny--
Sally Price
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Re: [Histonet] Histonet Digest, Vol 167, Issue 12

[Sally Price via Histonet]

All:
I beg to differ with what seems to be the general consensus, but the Dako
handbook is not really representative of the available technology within
the IHC field and shouldn't be thought of as a prescription for
success.  All the Dako book addresses is Dako's methods, reagents and
instruments, which is pretty biased.  In order to properly prepare for the
ASCPs exam, one needs to think beyond reagents and automated systems and
REALLY understand the underlying science.  I've attended a number of
seminars on this subject and learned that, especially because of recent
changes to this test, one also needs to understand the clinical application
of these procedures.  More and more we're called upon to provide
pathologists with support in the way of procedure validation and
troubleshooting and we need to learn a great deal more than can be learned
from reading one vendor's publication.
+Sally
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[Histonet] South Carolina Society for Histotechnology Fall Expo

[Sally Price via Histonet]

Hey folks –  I'm definitely not as good at promoting a meeting as Billie
Zimmerman is but I wouldn't want anyone to overlook their opportunity to
participate in the SCSH's Expo, which will be held the weekend of November
4 - 6 in beautiful Pawleys Island, just a short drive north of Charleston
and south of Myrtle Beach.  This may be your last chance to obtain
much-needed CEU's before the end of the year,  unless you enjoy those
boring on-line programs.  For more information, please contact SCSH's
President, Chad McMahan, who can be reached at < clmc...@clemson.edu >.
Hope to see y'all there!

-- 
Sally Price
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[Histonet] South Carolina Society for Histotechnology's Fall Expo

[Sally Price via Histonet]

Greetings HistoNetters - In the event that you couldn't make one of
the state-society meetings that were held in the Spring or the NSH
Symposium/Convention just a few weeks ago, I hope you'll consider attending
the South Carolina Society for Histotechnology's Fall Expo, which will be
held Friday, November 4th through Sunday, November 6th at the Litchfield
Beach and Golf Resort in Pawley's Island - which is right between Myrtle
Beach and Charleston.

Our meeting will feature nationally recognized speakers including Liz
Chlipala, B.S., HTL(ASCP), former South Carolinian Joe Myers, M.S.,
CT(ASCP)QIHC, and Bill DeSalvo, HT(ASCP) - who are not only knowledgeable
in their respective fields, but also quite entertaining...or so I'm told.

For more information on this event, please contact the SCSH's President,
Chad McMahan at: < clmc...@clemson.edu >

Hope to see ya'll there!
-- 
Sally Price
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Re: [Histonet] Tissue Controls (Deceptive)

[Sally Price via Histonet]

Not cool Rachel.  I took the survey assuming that it was intended and might 
help a colleage, but it's nothing but a marketing ploy, which are strongly 
discouraged on this forum.

 Original message 
From histonet-requ...@lists.utsouthwestern.edu 
Date: 06/15/2016  1:00 PM  (GMT-05:00) 
To histonet@lists.utsouthwestern.edu 
Subject Histonet Digest, Vol 151, Issue 14 
 
-

Message: 1
Date: Tue, 14 Jun 2016 14:09:16 -0400
From: Rachel M Gonzalez 
To: "histonet@lists.utsouthwestern.edu"

Subject: [Histonet] Tissue controls
Message-ID:

[Histonet] 12ml tubes for Dako IHC stainers

[Sally Price]

Can anyone recommend a vendor for the 12ml tubes that are used on original
Dako stainers other than Dako?



-- 
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[Histonet] Collagen fragmentation in skin sections after heat retrieval

[Sally Price]

Our lab has been experiencing a problem recently and we could use some
input on what might be causing it and what we might be able to do to
prevent it.  The issue is a very noticeable fragmentation of the dermal
collagen within most skin specimens, which isn't visible after HE
staining, but is after heat-retrieval.  We think our retrieval is pretty
gentle: It's performed in a veggie steamer (approx. 92oC) for 40 min,
followed by 15 min colloing on the countertop before rinsing.  We've tried
a variety of charged slides to see if the collagen holds on better, but it
doesn't.  We think our processing is pretty routine as well.  If it will
help, I can send a photo of this phenomenon to anyone who's interested.
Thanks,
Sally
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[Histonet] In FL, who can perform IHC/ISH?

[Sally Price]

I realize that this is a controversial topic, bu t I'm looking for a
definitive answer to the question of who can perfrom IHC and ISH in
Florida.  I ask this because, in addition to attempting to comply with the
Federal CLIA regs governing labs, Florida has personnel licensure regs that
cover our staff.  So, can an ASCP-registered technician (HT, MLT) handle
these procedures, or does it require a technologist (HTL, MT)?  And what
about unregistered/unlicensed people who work in physicians-office labs?
I'm especially interested in receiving feedback from the board members of
the FL Society for Histotechnology and respresentatives from the FL
Department of Health's division on Clinical Lab Personnel.
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[Histonet] New KOH-type stain

[Sally Price]

Histonetters:
The other day I recieved by mail a brochure for a stain in a one-gallon
bottle that claimed it stained fungal elements bluish-purple and
helped reading KOH's.  I gave the brochure to someone and they seem to have
misplaced it.  It wasn't the Chicago Sky Blue stain that's in those kits
(and is very expensive, 1 oz. bottles for $70).  Has anyone else received
this info and if so could they share the vendor/product info?
Cheers,
Sally
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RE: [Histonet] NCCI policy on IHC billing

[Sally Price]

Histonetters:
I waited a few days to see how others might weigh in after this information
was posted.  Call me crazy, but I expected quite a bit more reaction from
our community.  How is it that such a signifcant change in how IHC testing
may be conducted and will be paid for in the future can produce so little
response?

The way this new policy is stated, it looks pretty straightfoward: one
antibody (IHC procedure) per specimen; so, when it's necessary to use a
battery of IHC stains to determine the origin of an undifferentiated
neoplasm, the lab can only bill for one procedure.  How could such an
approach be possible?  And what about multi-antibody procedures, which
are usually more cost effective than single-antibody procedures?

Come on folks, this is a big deal becuase IHC staining is essential to to
the practice of anatomic pathology and provides a lot of us with our
livelihood.  I know I'm not alone in thinking that the CMS needs to know
that this new policy is completely impractical and must be changed.  Sure,
there's some unnecessary IHC procedures being performed, but this isn't the
way limit the problem.
Sally
--

Message: 6
Date: Fri, 30 Dec 2011 12:33:17 -0600
From: Webb, Dorothy L dorothy.l.w...@healthpartners.com
Subject: [Histonet] NCCI policy update
To: 'histonet@lists.utsouthwestern.edu'

Is everyone aware that beginning 1/1/12, we can no longer bill for each
block regarding IHC billing, only one unit of billing for each part type no
matter how many blocks are stained? Also IHC cocktail stains, such as
PIN4 must now be billed as one unit even though multiple antibodies are
reported out.

Kind of a surprising reversal of the policy set in motion 10/1/2009.
SPECIMEN becomes the unit of service rather than block(s) for IHC codes
88342, 88360, and 88361.

Happy New Year to everyone out there. May 2012 find you happiness and
health!

Dorothy Webb, HT
Regions Histology TS
651-254-2962
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[Histonet] p53 staining in mouse tissue

[Sally Price]

Dear HistoNetters:
I'm looking to identify p53 in mouse tissues and was hoping that you all
might be able to recommend sources of antibodies that react well in murine
tissues.
Thanks,
Sally
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RE: [Histonet] IHC validation

[Sally Price]

Liz:
I'd like to know more about these recommendations.  Could you provide a
journal reference to the paper from CAP?
Thx,
Sally

--

Message: 15
Date: Tue, 8 Feb 2011 16:19:53 -0700
From: Liz Chlipala l...@premierlab.com
Subject: RE: [Histonet] IHC validation
To: Weems, Joyce jwe...@sjha.org, Joe Nocito
jnoc...@satx.rr.com,Histonet histonet@lists.utsouthwestern.edu
Nope that's the recommendation for everything, in the paper they state
additional development is required for prognostic markers. Once you have
validated an antibody it only requires 3 tissues when you get a new lot
number:  1 strong positive, 1 weak to moderate positive and 1 negative.
From how I read the article its 25 tissues and then 3 tissues, it does
not talk about slides so it is possible to put multiple tissues on one
slide.  Again these are just recommendations; I do not think that there
is a set standard yet.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org]
Sent: Tuesday, February 08, 2011 4:13 PM
To: Liz Chlipala; Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

But that is for receptors, correct? Do you do that for everything?
Thanks, j

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz
Chlipala
Sent: Tuesday, February 08, 2011 17:58
To: Joe Nocito; Histonet
Subject: RE: [Histonet] IHC validation

Joe

If you are following the recommendations from the CAP paper on IHC
standardization then it would be 25 tissues (10 strong positive, 10 weak
to moderate positive and 5 negative).

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joe Nocito
Sent: Tuesday, February 08, 2011 3:44 PM
To: Histonet
Subject: [Histonet] IHC validation

Greetings Histoland,
I need some help. We are about to switch IHC machines from the
Richard-Allen Axiom to the Ventana Benchmark Ultra. How many slides, per
antibody, do you run for the validation study? We have over 100 primary
antibodies. Normally, when we work up a new antibody, we  start with a
titer. Once that is established, we run 10 cases to check for
specificity. Hopefully we can obtain cases that are really positive,
some weakly positive and some flat out negative. Once that is completed,
we run 10 different tissue types to check for any unexpected
cross-reactivity.
The ultra holds 30 slides and we are receiving two machines. If we
run 10 slides/antibody, that's going to take a while, not to mention the
number of detection kits that will be used. Do you think 5
slides/antibody is sufficient? I emailed CAP last week for their take
and they never returned my email (I told my medical director to hold
their check for the year and see how fast they respond to that). Ah oh,
don't go down that road Joe, it's unhealthy. What are your thoughts?
Thanks

Joe (JTT)
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RE: [Histonet] in situ - thanks 1 more query

[Sally Price]

Our anti-Dig reagent is included in Biocare's ISH detction kit. Its
unlabled, but is detected with a secondary-polymer-HRP reagent
that's directed at the anti-Dig primary.

--

Message: 14
Date: Tue, 25 Jan 2011 08:46:47 +0200
From: louise renton louise.ren...@gmail.com
Subject: [Histonet] in situ - thanks  1 more query
To: Histonet Histonet@lists.utsouthwestern.edu

Hi all, just wanted to say thanks for all the info received. L
Looking through the protocols it appears that i need an anti-DIG antibody,
preferably HRP labelled - any suggestions as to supplier? I remember using
one from DAKO, but don't see it in the latest catalogue. Please - any
sugestions as to where I can get it?

Thanks again - you guys (and gals) rock!!
--
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
+27 11 717 2298 (tel  fax)
073 5574456 (emergencies only)
There are nights when the wolves are silent and only the moon howls.
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
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[Histonet] RE: Help with Biocare polymer detection kit!!

[Sally Price]

Jennifer:
There are a variety possible causes for the background staining, which
probably have more to do with the specimen you're attempting to stain than
the detection systems, especially because you stated that you observed
similar background with a biotin/avidin-based system.  The first thing that
comes to mind is whether or not Mach 1 is intended for use on canine/feline
tissue; I suspect that its not, but you should check with Biocare's tech
support to be sure.  My guess is that either the immunologic enhancer
reagent or antibody conjugated to the polymer-enzyme reagent is binding to
endogenous immunolglobulins.  Another possible cause of the background is
that your tissue is over- retrieved/digested.  It could also be that your
antibody is not diluted adequately, or, as you suggested, the
protein-blocking reagent is actually contributing to the problem.
Good Luck!
Sally
--

Message: 1
Date: Sat, 31 Jul 2010 12:03:39 -0700
From: Jennifer Campbell jcampb...@vdxpathology.com
Subject: [Histonet] Help with Biocare polymer detection kit!!
To: histonet@lists.utsouthwestern.edu

I have been trying to get the Mach 1 polymer kit from Biocare to work on my
K9 and feline tissue and I am getting terrible nonspecific staining in my
positive and negative controls.  It appears to be sticking to something in
the glands of the intestinal epithelia.  I am also getting nonspecific
staining in my LN tissue.  I decided to switch over to polymer detection kit
because I had been dealing with the issue of endogenous biotin when using
the streptavidin biotin-based detection system, and now I am getting this
other nonspecific staining!  I have tried doing runs eliminating my protein
block and eliminating the negative control and primary antibody step
altogether (to see if maybe the protein block or the diluent used in my
negative control and antibody dilutions are causing the problem).  I have
also reduced the incubation time of both the probe and polymer from 15 min
and 30 min, respectively, to 10 min each.  I have tried cutting down my
antigen retrieval time substantially as well.

I'm not sure what else could be causing this.  Any suggestions??

Thanks in advance,

Jennifer Campbell
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[Histonet] Best charged slides for IHC

[Sally Price]

Has anybody done a comparison of charged slides, to see which ones are best
for IHC staining - espcially when slides will be pretreated in a pressure
cooker?
-- 
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[Histonet] RE: GI, Uro, or Derm Path Lab Set Up

[Sally Price]

...and another thing -- since when is it OK to post such blatant
solicitations on the HistoNet?  other vendors are chastised for this on a
regular basis!
Sally
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[Histonet] Ventana Her2/neu

[Sally Price]

On behalf of a colleaugue who doesn't participate in the Histonet, I'd like
to ask folks if there experiencing difficulty validating their Pathyway
Her2/neu antibody.  My friend has tried two different lots and both are
showing unusually weak staining.  Just wondering if anyone else is having
this issue and if you can offer some suggestions.
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[Histonet] Endogenous biotin blocking

[Sally Price]

I recently had a discussion with one of my coworkers about
the need/requirement for blocking of endoegnous biotin whenever an
avidin-biotin detection system is used, and I was hoping that the IHC
experts on the histonet might be able to provide us with some feeback.  Its
been my understanding that blocking is only necessary when one is certain
that background staining is caused by endogenous biotin, but maybe I'm
off-base here.
I look forward to eveyone's input.
Sally
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RE: [Histonet] Double HRP immunostains

[Sally Price]

Andrea - We love Biocare's Bajoran Purple.  It's out of this world...or it
galaxy?
 Sally

--

Message: 7
Date: Wed, 26 Aug 2009 14:05:41 -0400
From: Andrea Hooper anh2...@med.cornell.edu
Subject: [Histonet] Double HRP immunostains
To: Histonet histonet@lists.utsouthwestern.edu

I do double HRP immunostains. Works well using DAB/AEC and True Blue... but
what are your favorite chromagens to use? Would love some feedback for
inspiration.

Andrea
--
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RE: [Histonet] RE:Antigen retrieval paraffin skin samples

[Sally Price]

Maybe I've misinterpreted Nick's posting, but I think what he's looking for
is what kinds of reagents are used for retrieving Ck antigens.  It may
something to do with the PFA fixation, but I'm not too sure about using
ficin for CKs,  We usually use other enzymes or heat.  Proteinase K, or
trypsin, or citrate solutions work really well in most cases.
Sally
--

Message: 8
Date: Thu, 30 Jul 2009 09:30:37 -0600
From: Sorenson, Jon (Nampa) jonsoren...@chiwest.com
Subject: [Histonet] RE:Antigen retrieval paraffin skin samples
To: histonet@lists.utsouthwestern.edu

We use and like the PASCAL pressure cooker system from DAKO, it is uniform,
convenient and fairly Easy to use.
Jon Sorenson
Histology Coordinator
Mercy Medical Center
jonsore...@chiwest.com
208-463-5267

--

Message: 17
Date: Wed, 29 Jul 2009 14:05:29 -0700 (PDT)
From: Nicholas David Evans ndev...@stanford.edu
Subject: [HISTONET] Antigen retrieval paraffin skin samples
To: histonet@lists.utsouthwestern.edu

Dear all,
Might anyone be able to offer their advice on the best method to retrieve
antigens on paraffin embedded skin sections? I am immunostaining for
epithelial cytokeratins, and I have block-to-block variation in the quality
of the staining - some block give consistently good staining while others
giving negligible staining. The tissues were prepared by fixation in 4% PFA
for 24 - 48 hours followed by dehydration and embedding. I
currently use 15 mins Ficin digestion. I suspect that the differences may be
accounted for by variability in the length of time samples were fixed in PFA
and then 70% ethanol.
Best wishes
Nick
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RE: [Histonet] immunohistochemistry

[Sally Price]

Hatem:
Since this info hasn't already been offered, I'd suggest that you
considering using a 'programmable' water bath or pressure cooker -- where
the temperature is reduced to 65 to 75 degrees, but the exposure time
extended to three to six hours.
Sally

---

Date: Thu, 4 Jun 2009 20:01:50 -0700 (PDT)
From: Hatem Salim dr.hatemsa...@yahoo.com
Subject: [Histonet] immunohistochemistry
To: histonet@lists.utsouthwestern.edu

HI
I am Research assistant at Physiology department of Michigan state
university. I am using the (ß-Catenin Antibody (Carboxy-terminal Antigen)
#9587 for IHC on mice femur sections. I have used this antigen unmasking
method: For Citrate: Bring slides to a boil in 10 mM sodium citrate buffer
pH 6.0 then maintain at a sub-boiling temperature for 10 minutes. Cool
slides on bench top for 30 minutes.
The problem is that bone detachment always occurs so I wonder if there is a
method of retrieval that prevents bone detachment.
Waiting to hear from you soon.
Thank you for your consideration
Best wishes
Hatem Salim
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[Histonet] Need refurbished HE stainer

[Sally Price]

Fellow 'Netters:
I'm helping a friend set up a new lab and they'd like to get a refurbished
HE stainer.  Any suggestions, including likely price range, will be greatly
appreciated.  The lab's located in Ohio, if that makes a difference to
potential sellers.
Thx,
Sally
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RE: [Histonet] New Dako IHC Instrument and T.P.I.D.

[Sally Price]

Beth:
I evaluated the AS-Link/TPID and wasn't too impressed.  Its the same basic
machine with a new outside/skin and the only difference is that in can read
barcode labels on the reagents.  To me, the postive-ID part should be
handles by the LIS, not the staining system.  One part of the deal that
really concerned me is that Dako is pushing a new detectionsystem called
Flex, which costs a lot more than Envision plus. And they want to bill our
lab on a per-slide basis, which seems fishy to me.  Ican send you my cost
analysis if you want.  Next stop: Biocare's intellipath.
Sally
-Original Message-

From: histonet-boun...@lists.utsouthwestern.edu [mailto:
histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Fye Beth
Sent: Thursday, January 15, 2009 12:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Dako IHC Instrument and T.P.I.D.

I'm  curious if anyone out there has any feedback or comments on the new
Dako IHC Instrument, Autostainer Link 48 and/or their TPID (True Positive
ID).  We are in desparate need of replacing our current DAKO instruments,
and are probably going to stick with DAKO again after looking at several
other instruments.  I just thought I check to see if anyone using it has
anything to say.

Beth
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[Histonet] RE: Dako and Leica immunostainers

[Sally Price]

All,
After reading this thread I just had offer my comments.  I'm not a big fan
of systems that do the dewax and AR, primarily because it costs way too much
to automate these steps.  I've never used the Bond, but I hear that you've
gotta put some plastic thingy - that probably costs too much - on top of
each slide, you gotta use their detection reagents - which probably
cost more than other companies, they charge you for empty barcoded
reagent containers, all the slides in the same tray have to use the same
detection reagents - which means that the continuous-feed feature has some
serious limits, it can't do double-stains, and they have less than 50
IVD-approved antbodies.  Can someone verify for me if all this issues are
true?  If so, why would someone want one of these stainers?  The Dako
stainer is a dinosaur and with all the newre/better ones available,
they should probably take it off the market.
Cheers,
Sally

-Original Message-

From: Josie Britton [EMAIL PROTECTED]
To: Michael Bradley [EMAIL PROTECTED];
Histonet@lists.utsouthwestern.edu
Sent: Tue, 18 Nov 2008 11:31 am
Subject: RE: [Histonet] Dako

Hi,
We just got the new Bond Max IHC stainer and we love it.  You just cut the
slides dry them and place them on the Bond.  It does retrieval, antibody
staining, and counter stain.  You just dehydrate , clear, and mount your
coverslip.  It is easy to use.  It has 3 individual slide tray's of 10.  You
can load more slides on the empty tray's and start a new batch while the
others are running.  We run into the pathologist's adding more antibodies to
the list an hour after we have run the first batch frequently, so this
feature is great.  When you add more IHC's the run time on all the slide
tray's run times do increase, but it's better than having to wait another
2-3 hours to put your next set of immuno's on.
Hope this helps!
Josie Britton HT
Cheshire Medical Center
580 Court Street
Keene, NH 03431

-Original Message-

Message: 9
Date: Wed, 19 Nov 2008 17:17:52 -0500
From: [EMAIL PROTECTED]
Subject: Re: [Histonet] Dako
To: [EMAIL PROTECTED], [EMAIL PROTECTED],
Histonet@lists.utsouthwestern.edu

Just another FYI...
The Biocare IntelliPath does the same thing, but has 4 trays of 10 and it
allows you to make one of the trays?RUSH.? SO depending, on the volume?of
IHC you do either of these machines would work out great.
Roxanne
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