[Histonet] Re: uncertified techs in Histology (Chris Evanish)

Wed Feb 11 12:21:36 CST 2009

I have been in the Histology field for 20 years and I have seen
registered and nonresgistered techs and believe me some unregistered
tech did a better job then the registered. Passing a test and getting a
piece of paper does not mean that the tech was properly trained and has
experience and knowledge in the field. Plus now you have to go to
college to be eligible to take the test. 

>>> <histonet-request <@t> lists.utsouthwestern.edu> 2/11/2009 11:57 am >>>
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Today's Topics:

   1. Help with antigen unmaksing on frozen sections (Sarah Tarran)
   2. please unsubscribe (Cooperman, Sharon (NIH/NICHD) [E])
   3. RE: Sakura Glas Coverslipper and Xylene Sub (Breckenridge, Sue)
   4. semen analysis (Anne van Binsbergen)
   5. Re: Help with antigen unmaksing on frozen sections (Joanne
Mauger)
   6. Reagent Labeling Policy (Scott, Allison D)
   7. Re: Histonet Digest, Vol 63, Issue 13 (AWeiss <@t> shorememorial.org)
   8. uncertified techs in Histology (Steven Coakley)
   9. RE: uncertified techs in Histology (Pamela Marcum)
  10. brain sections falling off slides during IHC (Thurby, Christina)
  11. Re: uncertified techs in Histology (Rene J Buesa)
  12. Re: uncertified techs in Histology (Larry Woody)
  13. RE: brain sections falling off slides during IHC (Linke, Noelle)
  14. RE: uncertified techs in Histology (Mary Abosso)
  15. Re: uncertified techs in Histology (J. Fernandez)
  16. RE: brain sections falling off slides during IHC (Watson, Linda)
  17. RE: Sakura Glas Coverslipper and Xylene Sub (Breckenridge, Sue)
  18. RE: uncertified techs in Histology (Watson, Linda)
  19. Re: brain sections falling off slides during IHC (TF)
  20. RE: uncertified techs in Histology (Tom McNemar)
  21. Re: Help with antigen unmaksing on frozen sections ( TF )
  22. Subject: [Histonet] brain sections falling off slides during
      IHC (Bull, Jennifer L.)
  23. RE: uncertified techs in Histology (Jennifer MacDonald)

----------------------------------------------------------------------

Message: 1
Date: Wed, 11 Feb 2009 14:08:35 +1100 (EST)
From: "Sarah Tarran" <sarah_tarran <@t> wmi.usyd.edu.au>
Subject: [Histonet] Help with antigen unmaksing on frozen sections
To: histonet <@t> lists.utsouthwestern.edu 
Message-ID:
	<34319.192.195.170.5.1234321715.squirrel <@t> www.wmi.usyd.edu.au>
Content-Type: text/plain;charset=iso-8859-1

Hi everyone,

I some help with antigen on frozen sections as I am having a lot of
trouble getting antibodies to work. I am using a GFP mouse so can't
formalin fix, parafin embed my samples. I tried frsh freezing but
found
that the GFP leaked so I couldn't use that either. So I currently fix
my
tissue in 4% paraformaldehyde for 2 hours, then cryopreserve in 18%
sucrose overnight before embedding in OCT the following day. I have
tried
non-enzymatic heat retieval (boiling for 15 minutes in Dako's Target
retrieval solution pH 9) but this destroys the GFP, if I use proteinase
K
it 'eats' away at the tissue. I have tried diluting it but that hasn't
helped get the antibodies to work. I have also tried overnight at 4
degree
incubations without any luck.

Does anyone have any other ideas that I can try to get the antibodies
to
work? The antibodies that I am having problems with are: alpha smooth
muscle actin, LSP-1, procollagen, serum amyloid p and TGF-beta.

Thanks

Sarah Tarran
Postdoctoral Fellow
Vascular Biology Research Centre,
Department of Surgery
Westmead Hospital, Westmead, NSW, 2145

------------------------------

Message: 2
Date: Wed, 11 Feb 2009 00:14:53 -0500
From: "Cooperman, Sharon (NIH/NICHD) [E]" <cooperms <@t> mail.nih.gov>
Subject: [Histonet] please unsubscribe
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<2B184EA4ADDB0D41845D8B3730B10F651DA995 <@t> NIHCESMLBX7.nih.gov>
Content-Type: text/plain;	charset="iso-8859-1"

Thanks!

------------------------------

Message: 3
Date: Wed, 11 Feb 2009 07:38:00 -0500
From: "Breckenridge, Sue" <sbreckenridge <@t> caperegional.com>
Subject: RE: [Histonet] Sakura Glas Coverslipper and Xylene Sub
To: "Paula Lucas" <plucas <@t> biopath.org>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4D95C24EC0E4C84787A6919F1165B25E015B24D3 <@t> btmhems01.BTHOSP.INT>
Content-Type: text/plain;	charset="us-ascii"

Paula, try Micro-Clear CS from Micron Environmental Industries (phone
703-458-2776, catalog number HC1250). I have been using it for almost
20
years now; since 9/97 on the Leica CV5000 coverslipper. It's
comparable
in drying time to other xylene substitutes; but with a much better
safety aspect (and I don't notice any odor). 
Susan Breckenridge, Histology Supervisor, BS, HTL(ASCP)
Cape Regional Medical Center
Cape May Court House, NJ
P.S. I am an end user of the product, not a sales rep. I do not get
any
compensation for my opinion. 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula
Lucas
Sent: Tuesday, February 10, 2009 1:41 PM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] Sakura Glas Coverslipper and Xylene Sub

Hello,

We use Xylene in the Sakura Glas auto-coverslipper and I would like to
know if anyone uses a xylene sub in this piece of equipment. I'm
looking
for one that is okay to use in the equipment and one with less odor
than
Xylene.

Thanks in advance,
Paula Lucas
Lab Manager
BioPath MG

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Message: 4
Date: Wed, 11 Feb 2009 16:45:48 +0400
From: Anne van Binsbergen <annigyg <@t> gmail.com>
Subject: [Histonet] semen analysis
To: histonet <@t> lists.utsouthwestern.edu 
Message-ID:
	<f8332fbe0902110445n64a7c363m38eb32804679e080 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

who out there is using automation for semen analysis and what equipment
is
recommended?
i am considering the SQA-V from MES
what say you?

cheers

-- 
Anne van Binsbergen (Hope)
Abu Dhabi
UAE

------------------------------

Message: 5
Date: Wed, 11 Feb 2009 08:17:24 -0500
From: "Joanne Mauger" <MAUGER <@t> email.chop.edu>
Subject: Re: [Histonet] Help with antigen unmaksing on frozen sections
To: <histonet <@t> lists.utsouthwestern.edu>,<sarah_tarran <@t> wmi.usyd.edu.au>
Message-ID: <s99289a7.007 <@t> email.chop.edu>
Content-Type: text/plain; charset=US-ASCII

Sarah,

Try fixing your cut sections in 10%NBF for 20-30 minutes after
deparaffinizing, then do the retrievals. SMA doesn't usually require
any
retrieval. TCR beta works well with pepsin for 20 mins. - this is less
harsh than proK. I'm not familiar with the other abs you mention.

Jo Mauger

>>> "Sarah Tarran" <sarah_tarran <@t> wmi.usyd.edu.au> 02/10/09 10:08 PM
>>>
Hi everyone,

I some help with antigen on frozen sections as I am having a lot of
trouble getting antibodies to work. I am using a GFP mouse so can't
formalin fix, parafin embed my samples. I tried frsh freezing but
found
that the GFP leaked so I couldn't use that either. So I currently fix
my
tissue in 4% paraformaldehyde for 2 hours, then cryopreserve in 18%
sucrose overnight before embedding in OCT the following day. I have
tried
non-enzymatic heat retieval (boiling for 15 minutes in Dako's Target
retrieval solution pH 9) but this destroys the GFP, if I use
proteinase
K
it 'eats' away at the tissue. I have tried diluting it but that hasn't
helped get the antibodies to work. I have also tried overnight at 4
degree
incubations without any luck.

Does anyone have any other ideas that I can try to get the antibodies
to
work? The antibodies that I am having problems with are: alpha smooth
muscle actin, LSP-1, procollagen, serum amyloid p and TGF-beta.

Thanks

Sarah Tarran
Postdoctoral Fellow
Vascular Biology Research Centre,
Department of Surgery
Westmead Hospital, Westmead, NSW, 2145

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 6
Date: Wed, 11 Feb 2009 07:24:39 -0600
From: "Scott, Allison D" <Allison_Scott <@t> hchd.tmc.edu>
Subject: [Histonet] Reagent Labeling Policy
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<1872B4A455B7974391609AD8034C79FC8BD503 <@t> LBEXCH01.hchd.local>
Content-Type: text/plain; charset="us-ascii"

Does anyone have a reagent labeling policy that they would be willing
to
share.  Thanks in advance

Allison Scott HT(ASCP)
Histology Supervisor
LBJ Hospital
Houston, Texas 77026
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------------------------------

Message: 7
Date: Wed, 11 Feb 2009 08:34:06 -0500
From: AWeiss <@t> shorememorial.org 
Subject: [Histonet] Re: Histonet Digest, Vol 63, Issue 13
To: histonet <@t> lists.utsouthwestern.edu 
Message-ID:
	<OF13E03C62.3791FEEE-ON8525755A.004A66A7-8525755A.004AA61E <@t> shorememorial.org>

Content-Type: text/plain; charset=ISO-8859-1

I do FNA biospy and adequacy at bedside with radiologists and
pulmonary
docs I use 88173  this includes tech component.

Is anyone aware of a CPT code for a Tech assisted bone marrow
aspiration
and biopsy?  From time to time we have been helping the Oncologist and
Radiologist on these procedures. It is becoming more of a common
practice
and we spend anywhere from a ½ hour to 1 ½ hours and not billing for
our
services. Is there anyone else in a similar situation?

Andrea J Weiss BST CT (ASCP)
Cytotechnologist
609 653 3577 Ext 4907
aweiss <@t> shorememorial.org 

------------------------------

Message: 8
Date: Wed, 11 Feb 2009 06:55:56 -0800 (PST)
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] uncertified techs in Histology
To: Histonet <@t> lists.utsouthwestern.edu 
Message-ID: <816453.11897.qm <@t> web38208.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big advantage do all these facilities think there gaining by going with
unregistered techs, especially when theres always ongoing quality issues
when theres so many trained certified HT looking for work?  In my area
of the country I can't believe how many Hospitals go this way.

------------------------------

Message: 9
Date: Wed, 11 Feb 2009 10:21:52 -0500
From: "Pamela Marcum" <pmarcum <@t> vet.upenn.edu>
Subject: RE: [Histonet] uncertified techs in Histology
To: "'Steven Coakley'" <sjchtascp <@t> yahoo.com>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001501c98c5c$77d92850$095a5b82 <@t> vet.upenn.edu>
Content-Type: text/plain;	charset="iso-8859-1"

It is called be cheap and don't pay for training!!  Sorry that is how I
see
and ASCP does not seem to care or force the issue for training in
histology.

Pamela A Marcum
University of Pennsylvania 
School of Veterinary Medicine
Comparative Orthopedic Laboratory (CORL)
382 W Street Rd
Kennett Square PA 19438
610-925-6278

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Steven
Coakley
Sent: Wednesday, February 11, 2009 9:56 AM
To: Histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] uncertified techs in Histology

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
advantage do all these facilities think there gaining by going with
unregistered techs, especially when theres always ongoing quality
issues
when theres so many trained certified HT looking for work?  In my area
of
the country I can't believe how many Hospitals go this way.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 10
Date: Wed, 11 Feb 2009 10:37:55 -0500
From: "Thurby, Christina" <christina.thurby <@t> bms.com>
Subject: [Histonet] brain sections falling off slides during IHC
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<E9C77491626EBF41BD449EA1098B04062749DB70 <@t> ushpwbmsmmp007.one.ads.bms.com>

Content-Type: text/plain; charset="us-ascii"

Hello all,
Can any of you all with experience immunostaining brain sections please
help.  My sections are almost completely washing off the slides.  I am
using 10% NBF fixed canine brain sections.  Embedded in paraffin and
sectioned at 4 microns on plus slides with distilled water in the water
bath.  The slides are dried at 65 C for 15 minutes and allowed to stand
overnight.  I am using a Dako Autostainer for staining.  There is no
antigen retrieval step.  My staining procedure is approximately 3.5
hours long.  At the completion of the procedure, the tissue on my slides
is 50-80% washed off.

Any thoughts??
Christina Thurby

------------------------------

Message: 11
Date: Wed, 11 Feb 2009 07:58:26 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] uncertified techs in Histology
To: Histonet <@t> lists.utsouthwestern.edu, Steven Coakley
	<sjchtascp <@t> yahoo.com>
Message-ID: <283333.28525.qm <@t> web65715.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

A hospital that relies on uncertified techs to do histology work is
motivated by the pursue of costs cuts (you can call it greed!) and shows
total disregard for quality of work and patient care. They may end
losing all those savings when settling a legal case.
René J.

--- On Wed, 2/11/09, Steven Coakley <sjchtascp <@t> yahoo.com> wrote:

From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] uncertified techs in Histology
To: Histonet <@t> lists.utsouthwestern.edu 
Date: Wednesday, February 11, 2009, 9:55 AM

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
advantage do all these facilities think there gaining by going with
unregistered
techs, especially when theres always ongoing quality issues when theres
so many
trained certified HT looking for work?  In my area of the country I
can't
believe how many Hospitals go this way.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 12
Date: Wed, 11 Feb 2009 08:09:39 -0800 (PST)
From: Larry Woody <slappycraw <@t> yahoo.com>
Subject: Re: [Histonet] uncertified techs in Histology
To: rjbuesa <@t> yahoo.com, Histonet <@t> lists.utsouthwestern.edu,	Steven
	Coakley <sjchtascp <@t> yahoo.com>
Message-ID: <948599.42650.qm <@t> web53610.mail.re2.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

This has been an ongoing issue for so many years in histology, I've
always wanted to see a mandatory license in the field but that always
starts a firestorm of controversy. If you have surgery, you certainly
want a board certified surgeon to do it and same with the Pathologist
that looks at the slides so wouldn't you want a certified tech doing the
lab work as well?

Larry A. Woody
Seattle, Wa.

________________________________
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
To: Histonet <@t> lists.utsouthwestern.edu; Steven Coakley
<sjchtascp <@t> yahoo.com>
Sent: Wednesday, February 11, 2009 7:58:26 AM
Subject: Re: [Histonet] uncertified techs in Histology

A hospital that relies on uncertified techs to do histology work is
motivated by the pursue of costs cuts (you can call it greed!) and shows
total disregard for quality of work and patient care. They may end
losing all those savings when settling a legal case.
René J.

--- On Wed, 2/11/09, Steven Coakley <sjchtascp <@t> yahoo.com> wrote:

From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] uncertified techs in Histology
To: Histonet <@t> lists.utsouthwestern.edu 
Date: Wednesday, February 11, 2009, 9:55 AM

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
advantage do all these facilities think there gaining by going with
unregistered
techs, especially when theres always ongoing quality issues when theres
so many
trained certified HT looking for work?  In my area of the country I
can't
believe how many Hospitals go this way.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 13
Date: Wed, 11 Feb 2009 08:11:43 -0800
From: "Linke, Noelle" <NLinke <@t> mednet.ucla.edu>
Subject: [Histonet] RE: brain sections falling off slides during IHC
To: "Thurby, Christina" <christina.thurby <@t> bms.com>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<0C96F0BFE078D74C91A1C541D24A6AE43D28AEE6 <@t> EMGMB1.ad.medctr.ucla.edu>
Content-Type: text/plain; charset="koi8-r"

Hi Christina,

In my experience, there are a few things to try.  First, don't soak
your blocks too long before cutting, and try not to let too much water
get underneath the sections as you pick them up from the waterbath.  I
wouldn't put them in an oven at all, just let them air dry as long as
you can (overnight, a day if you can spare the time) before staining.

If you're having troubles, I wouldn't use the autostainer at all (it
rinses too aggressively for some tissues), I would do them manually and
never let the stream from your wash bottle touch the tissue, gently
rinse after each step.

I hope this helps.

Noelle

No£lle Linke M.S., HTL(ASCP)QIHC
Manager, Histology Services
Department of Pathology & Laboratory Medicine
David Geffen School of Medicine at UCLA
Phone: 310-825-7397
Pager: 97471
nlinke <@t> mednet.ucla.edu 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Thurby, Christina
Sent: Wednesday, February 11, 2009 7:38 AM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] brain sections falling off slides during IHC

Hello all,
Can any of you all with experience immunostaining brain sections please
help.  My sections are almost completely washing off the slides.  I am
using 10% NBF fixed canine brain sections.  Embedded in paraffin and
sectioned at 4 microns on plus slides with distilled water in the water
bath.  The slides are dried at 65 C for 15 minutes and allowed to stand
overnight.  I am using a Dako Autostainer for staining.  There is no
antigen retrieval step.  My staining procedure is approximately 3.5
hours long.  At the completion of the procedure, the tissue on my slides
is 50-80% washed off.

Any thoughts??
Christina Thurby
_______________________________________________
Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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------------------------------

Message: 14
Date: Wed, 11 Feb 2009 09:18:39 -0700
From: "Mary Abosso" <mabosso <@t> unipathllc.com>
Subject: RE: [Histonet] uncertified techs in Histology
To: "Steven Coakley" <sjchtascp <@t> yahoo.com>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<43A451981FF6634795BE83B1B5494D631BDF25 <@t> exchange.unipathllc.corp>
Content-Type: text/plain;	charset="iso-8859-1"

Many areas have to go to unregistered techs out of necessity due to the
high vacancy rate in our field since the ASCP made changes to the career
path for the HT board requirements.  Many of these facilities have
either formal or informal training and have turned out many very good,
high quality techs.  Yes, some may be skilled at the "art" of histology,
but lack the understanding that is behind the science.  These often can
embed, cut and do special stains with superior quality and happy
pathologists.  I have seen registered techs that have all the
intelligence that their sheepskins say, but totally lack the hands on
skills needed to produce superior quality material for patient
diagnosis.  While this is only my opinion, I wish that there was still
the route for OJT as long as there is a certified tech in place for
training.

My two cents worth,
Mary Abosso 

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven
Coakley
Sent: Wed 2/11/2009 7:55 AM
To: Histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] uncertified techs in Histology

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big advantage do all these facilities think there gaining by going with
unregistered techs, especially when theres always ongoing quality issues
when theres so many trained certified HT looking for work?  In my area
of the country I can't believe how many Hospitals go this way.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 15
Date: Wed, 11 Feb 2009 08:20:49 -0800 (PST)
From: "J. Fernandez" <dez <@t> u.washington.edu>
Subject: Re: [Histonet] uncertified techs in Histology
To: Steven Coakley <sjchtascp <@t> yahoo.com>
Cc: Histonet <@t> lists.utsouthwestern.edu 
Message-ID:
	<Pine.LNX.4.64.0902110816110.23147 <@t> homer21.u.washington.edu>
Content-Type: text/plain; charset="iso-8859-1"

What does quality issues have to do with anything ,Many certifieds have

quality issues as well,It's knowledge and skill that matters.You can go
on 
all day with this ,but in the end it's can you get it done,available
techs 
etc.....I think it's important to have the best of both worlds.

On Wed, 11 Feb 2009, Steven Coakley wrote:

> Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big advantage do all these facilities think there gaining by going with
unregistered techs, especially when theres always ongoing quality issues
when theres so many trained certified HT looking for work?  In my area
of the country I can't believe how many Hospitals go this way.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>

------------------------------

Message: 16
Date: Wed, 11 Feb 2009 11:24:48 -0500
From: "Watson, Linda" <Linda.Watson <@t> bms.com>
Subject: [Histonet] RE: brain sections falling off slides during IHC
To: "Linke, Noelle" <NLinke <@t> mednet.ucla.edu>, "Thurby, Christina"
	<christina.thurby <@t> bms.com>, "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<351A66CE7FB11D40AB8AC8FE5559EC7E01A0DBB9DD <@t> ushpwbmsmmp008.one.ads.bms.com>

Content-Type: text/plain; charset="koi8-r"

Just to add to Christina,
Try Gold Plus Slides-Thermo Shandon (previously Erie Scientific).

Linda

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Linke, Noelle
>Sent: Wednesday, February 11, 2009 11:12 AM
>To: Thurby, Christina; histonet <@t> lists.utsouthwestern.edu 
>Subject: [Histonet] RE: brain sections falling off slides during IHC
>
>Hi Christina,
>
>In my experience, there are a few things to try.  First, don't soak
your
>blocks too long before cutting, and try not to let too much water get
>underneath the sections as you pick them up from the waterbath.  I
>wouldn't put them in an oven at all, just let them air dry as long as
>you can (overnight, a day if you can spare the time) before staining.
>
>If you're having troubles, I wouldn't use the autostainer at all (it
>rinses too aggressively for some tissues), I would do them manually
and
>never let the stream from your wash bottle touch the tissue, gently
>rinse after each step.
>
>I hope this helps.
>
>Noelle
>
>No£lle Linke M.S., HTL(ASCP)QIHC
>Manager, Histology Services
>Department of Pathology & Laboratory Medicine
>David Geffen School of Medicine at UCLA
>Phone: 310-825-7397
>Pager: 97471
>nlinke <@t> mednet.ucla.edu 
>
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Thurby, Christina
>Sent: Wednesday, February 11, 2009 7:38 AM
>To: histonet <@t> lists.utsouthwestern.edu 
>Subject: [Histonet] brain sections falling off slides during IHC
>
>Hello all,
>Can any of you all with experience immunostaining brain sections
please
>help.  My sections are almost completely washing off the slides.  I
am
>using 10% NBF fixed canine brain sections.  Embedded in paraffin and
>sectioned at 4 microns on plus slides with distilled water in the
water
>bath.  The slides are dried at 65 C for 15 minutes and allowed to
stand
>overnight.  I am using a Dako Autostainer for staining.  There is no
>antigen retrieval step.  My staining procedure is approximately 3.5
>hours long.  At the completion of the procedure, the tissue on my
slides
>is 50-80% washed off.
>
>Any thoughts??
>Christina Thurby
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>
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------------------------------

Message: 17
Date: Wed, 11 Feb 2009 11:26:49 -0500
From: "Breckenridge, Sue" <sbreckenridge <@t> caperegional.com>
Subject: RE: [Histonet] Sakura Glas Coverslipper and Xylene Sub
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4D95C24EC0E4C84787A6919F1165B25E015B24D8 <@t> btmhems01.BTHOSP.INT>
Content-Type: text/plain;	charset="us-ascii"

Opps, sorry folks, had a dyslectic moment. The phone number is:
703-548-2776. My apologies for any confusion, Sue

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Breckenridge, Sue
Sent: Wednesday, February 11, 2009 7:38 AM
To: Paula Lucas; histonet <@t> lists.utsouthwestern.edu 
Subject: RE: [Histonet] Sakura Glas Coverslipper and Xylene Sub

Paula, try Micro-Clear CS from Micron Environmental Industries (phone
703-458-2776, catalog number HC1250). I have been using it for almost
20
years now; since 9/97 on the Leica CV5000 coverslipper. It's
comparable
in drying time to other xylene substitutes; but with a much better
safety aspect (and I don't notice any odor). 
Susan Breckenridge, Histology Supervisor, BS, HTL(ASCP)
Cape Regional Medical Center
Cape May Court House, NJ
P.S. I am an end user of the product, not a sales rep. I do not get
any
compensation for my opinion. 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula
Lucas
Sent: Tuesday, February 10, 2009 1:41 PM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] Sakura Glas Coverslipper and Xylene Sub

Hello,

We use Xylene in the Sakura Glas auto-coverslipper and I would like to
know if anyone uses a xylene sub in this piece of equipment. I'm
looking
for one that is okay to use in the equipment and one with less odor
than
Xylene.

Thanks in advance,
Paula Lucas
Lab Manager
BioPath MG

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Message: 18
Date: Wed, 11 Feb 2009 11:33:40 -0500
From: "Watson, Linda" <Linda.Watson <@t> bms.com>
Subject: RE: [Histonet] uncertified techs in Histology
To: Mary Abosso <mabosso <@t> unipathllc.com>, Steven Coakley
	<sjchtascp <@t> yahoo.com>, "Histonet <@t> lists.utsouthwestern.edu"
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<351A66CE7FB11D40AB8AC8FE5559EC7E01A0DBB9EC <@t> ushpwbmsmmp008.one.ads.bms.com>

Content-Type: text/plain; charset="us-ascii"

I totally agree with Mary. I have often heard that histotechs are a
dying breed. That us "old timers" are not being replaced by the younger
generation. If we want to have less uncertified histotechs then lets not
make it so difficult for individuals to pursue this wonderful career. I
still think it is very important for the exam to remain in place but
those individuals out of high school that are maybe not thinking of
attending college to obtain a bachelors degree should have the
opportunity to become a certified histologist. I know that you can still
obtain the HT but not the HTL. Correct me if I am wrong.

Linda

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mary Abosso
>Sent: Wednesday, February 11, 2009 11:19 AM
>To: Steven Coakley; Histonet <@t> lists.utsouthwestern.edu 
>Subject: RE: [Histonet] uncertified techs in Histology
>
>Many areas have to go to unregistered techs out of necessity due to
the
>high vacancy rate in our field since the ASCP made changes to the
career
>path for the HT board requirements.  Many of these facilities have
>either formal or informal training and have turned out many very
good,
>high quality techs.  Yes, some may be skilled at the "art" of
histology,
>but lack the understanding that is behind the science.  These often
can
>embed, cut and do special stains with superior quality and happy
>pathologists.  I have seen registered techs that have all the
>intelligence that their sheepskins say, but totally lack the hands on
>skills needed to produce superior quality material for patient
>diagnosis.  While this is only my opinion, I wish that there was
still
>the route for OJT as long as there is a certified tech in place for
>training.
>
>My two cents worth,
>Mary Abosso
>
>________________________________
>
>From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven
>Coakley
>Sent: Wed 2/11/2009 7:55 AM
>To: Histonet <@t> lists.utsouthwestern.edu 
>Subject: [Histonet] uncertified techs in Histology
>
>
>
>Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
>advantage do all these facilities think there gaining by going with
>unregistered techs, especially when theres always ongoing quality
issues
>when theres so many trained certified HT looking for work?  In my
area
>of the country I can't believe how many Hospitals go this way.
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 19
Date: Thu, 12 Feb 2009 00:34:38 +0800
From: "TF" <tifei <@t> foxmail.com>
Subject: Re: [Histonet] brain sections falling off slides during IHC
To: "Thurby, Christina" <christina.thurby <@t> bms.com>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <200902120034336211998 <@t> foxmail.com>
Content-Type: text/plain;	charset="gb2312"

Hi, 
my experience is to use lower concentration gel-coated slides or
pre-coated slide purchased from company...dont use home made one if you
want to improve.

second, the heating procedure shortly after mounting the frozen
sections onto slides does not help a lot. 
My way: mounting sections from cryostat onto slide -> air dry -> heat
plate 40-50 C or not for 10 min until the slides are a bit hot -> air
dry in fume hood overnight (for small section such as Olfactory bulb,
coronal brain section of rat takes 1-2 days - much better) ->you can
stock all sections in -20 C freezer ->each time before IHC, heat the
section at 60 C for 10 min (dry up after taking out from freezer).

I boil the slide in 95 C citrate buffer for 30 min, <1 sections from
100 falls off.

2009-02-12 

TF 

·¢¼þÈË£º Thurby, Christina 
·¢ËÍÊ±¼ä£º 2009-02-11  23:43:16 
ÊÕ¼þÈË£º histonet <@t> lists.utsouthwestern.edu 
³-ËÍ£º 
Ö÷Ìâ£º [Histonet] brain sections falling off slides during IHC 

Hello all,
Can any of you all with experience immunostaining brain sections please
help.  My sections are almost completely washing off the slides.  I am
using 10% NBF fixed canine brain sections.  Embedded in paraffin and
sectioned at 4 microns on plus slides with distilled water in the water
bath.  The slides are dried at 65 C for 15 minutes and allowed to stand
overnight.  I am using a Dako Autostainer for staining.  There is no
antigen retrieval step.  My staining procedure is approximately 3.5
hours long.  At the completion of the procedure, the tissue on my slides
is 50-80% washed off.
Any thoughts??
Christina Thurby
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 20
Date: Wed, 11 Feb 2009 11:36:02 -0500
From: "Tom McNemar" <TMcNemar <@t> lmhealth.org>
Subject: RE: [Histonet] uncertified techs in Histology
To: "Larry Woody" <slappycraw <@t> yahoo.com>, <rjbuesa <@t> yahoo.com>,
	<Histonet <@t> lists.utsouthwestern.edu>, "Steven Coakley"
	<sjchtascp <@t> yahoo.com>
Message-ID:
	<51D5D78FBEDAEA4FBCCD9A9D44211DC505B8E0A7 <@t> lmhsmail.lmhealth.org>
Content-Type: text/plain;	charset="iso-8859-1"

Perhaps in a perfect world....  My world is less than perfect.  For our
last opening, we spent 10 months trying to find and lure a certified
tech to our facility and then gave up and took an MLT.  We have four
techs and two of us are certified HTs.  We recently hired a person off
the street and trained them to be a histology assistant.  It has been
very beneficial for us.  She files slides, covers the late grossing
(assists the pathologist), coverslips, etc.

It doesn't take an advanced degree to do histology.  You gotta do what
you gotta do to get the work out.

Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcnemar <@t> lmhealth.org 
www.LMHealth.org 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Larry
Woody
Sent: Wednesday, February 11, 2009 11:10 AM
To: rjbuesa <@t> yahoo.com; Histonet <@t> lists.utsouthwestern.edu; Steven
Coakley
Subject: Re: [Histonet] uncertified techs in Histology

This has been an ongoing issue for so many years in histology, I've
always wanted to see a mandatory license in the field but that always
starts a firestorm of controversy. If you have surgery, you certainly
want a board certified surgeon to do it and same with the Pathologist
that looks at the slides so wouldn't you want a certified tech doing the
lab work as well?

Larry A. Woody
Seattle, Wa.

________________________________
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
To: Histonet <@t> lists.utsouthwestern.edu; Steven Coakley
<sjchtascp <@t> yahoo.com>
Sent: Wednesday, February 11, 2009 7:58:26 AM
Subject: Re: [Histonet] uncertified techs in Histology

A hospital that relies on uncertified techs to do histology work is
motivated by the pursue of costs cuts (you can call it greed!) and shows
total disregard for quality of work and patient care. They may end
losing all those savings when settling a legal case.
René J.

--- On Wed, 2/11/09, Steven Coakley <sjchtascp <@t> yahoo.com> wrote:

From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] uncertified techs in Histology
To: Histonet <@t> lists.utsouthwestern.edu 
Date: Wednesday, February 11, 2009, 9:55 AM

Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
advantage do all these facilities think there gaining by going with
unregistered
techs, especially when theres always ongoing quality issues when theres
so many
trained certified HT looking for work?  In my area of the country I
can't
believe how many Hospitals go this way.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

Message: 21
Date: Thu, 12 Feb 2009 00:36:16 +0800
From: " TF " <tifei <@t> foxmail.com>
Subject: Re: [Histonet] Help with antigen unmaksing on frozen sections
To: " Sarah Tarran " <sarah_tarran <@t> wmi.usyd.edu.au>, " histonet "
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <200902120036112922997 <@t> foxmail.com>
Content-Type: text/plain;	charset="utf-8"

hi, for alpha -SMA
I just put the slide with section into a plastic jar with citrate
buffer
put the jar into microwave, "HIGH" for 1 min and quickly cool down by
adding cool distilled water
then use 0.3% trition-PBS solution to soak the section for 10 min.
works very well.

i can show u pictures if u want,.

2009-02-12 

TF 

å ‘ä»¶äººï¼š Sarah Tarran 
å ‘é€ æ—¶é—́ï¼š 2009-02-11  12:19:58 
æ”¶ä»¶äººï¼š histonet 
æŠ„é€ ï¼š 
ä̧»é¢̃ï¼š [Histonet] Help with antigen unmaksing on frozen sections 

Hi everyone,
I some help with antigen on frozen sections as I am having a lot of
trouble getting antibodies to work. I am using a GFP mouse so can't
formalin fix, parafin embed my samples. I tried frsh freezing but
found
that the GFP leaked so I couldn't use that either. So I currently fix
my
tissue in 4% paraformaldehyde for 2 hours, then cryopreserve in 18%
sucrose overnight before embedding in OCT the following day. I have
tried
non-enzymatic heat retieval (boiling for 15 minutes in Dako's Target
retrieval solution pH 9) but this destroys the GFP, if I use proteinase
K
it 'eats' away at the tissue. I have tried diluting it but that hasn't
helped get the antibodies to work. I have also tried overnight at 4
degree
incubations without any luck.
Does anyone have any other ideas that I can try to get the antibodies
to
work? The antibodies that I am having problems with are: alpha smooth
muscle actin, LSP-1, procollagen, serum amyloid p and TGF-beta.
Thanks
Sarah Tarran
Postdoctoral Fellow
Vascular Biology Research Centre,
Department of Surgery
Westmead Hospital, Westmead, NSW, 2145
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------------------------------

Message: 22
Date: Wed, 11 Feb 2009 08:38:49 -0800
From: "Bull, Jennifer L." <Jennifer.Bull <@t> northwestpathology.com>
Subject: Subject: [Histonet] brain sections falling off slides during
	IHC
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<85760CECEC18444BB95F26D5E88DAEAA21FF216F6F <@t> hinet2.hinet.org>
Content-Type: text/plain; charset="us-ascii"

What  type of plus slides are you using? There are a few companies out
there selling a "super" adhesive slide for just such applications. We
use the Bond380 slide from TruScientific for all of our brain, autopsy
and fatty tissue when performing IHC. We rarely have any compromised
tissue and that is using the Dako platform and the PT Module for
retrieval. I know there are other companies that carry similar slides,
but we found the best pricing on these. You can reach them at
877-677-7523.

You might also try skipping the oven and air drying your sections
overnight follwed by a run over the heat plate before starting the
staining process.

Jennifer Bull
jennifer.bull <@t> nwpathology.com 

-----Original Message-----

From: histonet-bounces <@t>
lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet>
[mailto:histonet-bounces <@t>
lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet>]
On Behalf Of Thurby, Christina

Sent: Wednesday, February 11, 2009 7:38 AM

To: histonet <@t>
lists.utsouthwestern.edu<http://lists.utsouthwestern.edu/mailman/listinfo/histonet>

Subject: [Histonet] brain sections falling off slides during IHC

Hello all,

Can any of you all with experience immunostaining brain sections please
help.  My sections are almost completely washing off the slides.  I am
using 10% NBF fixed canine brain sections.  Embedded in paraffin and
sectioned at 4 microns on plus slides with distilled water in the water
bath.  The slides are dried at 65 C for 15 minutes and allowed to stand
overnight.  I am using a Dako Autostainer for staining.  There is no
antigen retrieval step.  My staining procedure is approximately 3.5
hours long.  At the completion of the procedure, the tissue on my slides
is 50-80% washed off.

Any thoughts??

Christina Thurby

------------------------------

Message: 23
Date: Wed, 11 Feb 2009 08:51:47 -0800
From: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>
Subject: RE: [Histonet] uncertified techs in Histology
To: "Watson, Linda" <Linda.Watson <@t> bms.com>
Cc: "Histonet <@t> lists.utsouthwestern.edu"
	<Histonet <@t> lists.utsouthwestern.edu>,
	histonet-bounces <@t> lists.utsouthwestern.edu 
Message-ID:
	<OF93B4BB2B.285E6873-ON8825755A.005C6F4A-8825755A.005CD8C5 <@t> mtsac.edu>
Content-Type: text/plain; charset="US-ASCII"

Both the HT and the HTL are available to qualified individuals.  The
only 
thing that was discontinued was the High School or equivalent and 2
years 
of experience (route 3).  All other options (routes) remain the same.

Jennifer MacDonald
Mt. San Antonio College

"Watson, Linda" <Linda.Watson <@t> bms.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu 
02/11/2009 08:36 AM

To
Mary Abosso <mabosso <@t> unipathllc.com>, Steven Coakley 
<sjchtascp <@t> yahoo.com>, "Histonet <@t> lists.utsouthwestern.edu" 
<Histonet <@t> lists.utsouthwestern.edu>
cc

Subject
RE: [Histonet] uncertified techs in Histology

I totally agree with Mary. I have often heard that histotechs are a
dying 
breed. That us "old timers" are not being replaced by the younger 
generation. If we want to have less uncertified histotechs then lets
not 
make it so difficult for individuals to pursue this wonderful career. I

still think it is very important for the exam to remain in place but
those 
individuals out of high school that are maybe not thinking of attending

college to obtain a bachelors degree should have the opportunity to
become 
a certified histologist. I know that you can still obtain the HT but
not 
the HTL. Correct me if I am wrong.

Linda

>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-
>bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mary Abosso
>Sent: Wednesday, February 11, 2009 11:19 AM
>To: Steven Coakley; Histonet <@t> lists.utsouthwestern.edu 
>Subject: RE: [Histonet] uncertified techs in Histology
>
>Many areas have to go to unregistered techs out of necessity due to
the
>high vacancy rate in our field since the ASCP made changes to the
career
>path for the HT board requirements.  Many of these facilities have
>either formal or informal training and have turned out many very
good,
>high quality techs.  Yes, some may be skilled at the "art" of
histology,
>but lack the understanding that is behind the science.  These often
can
>embed, cut and do special stains with superior quality and happy
>pathologists.  I have seen registered techs that have all the
>intelligence that their sheepskins say, but totally lack the hands on
>skills needed to produce superior quality material for patient
>diagnosis.  While this is only my opinion, I wish that there was
still
>the route for OJT as long as there is a certified tech in place for
>training.
>
>My two cents worth,
>Mary Abosso
>
>________________________________
>
>From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Steven
>Coakley
>Sent: Wed 2/11/2009 7:55 AM
>To: Histonet <@t> lists.utsouthwestern.edu 
>Subject: [Histonet] uncertified techs in Histology
>
>
>
>Any thoughts or experiences with my fellow HT/HTL's(ASCP).  What the
big
>advantage do all these facilities think there gaining by going with
>unregistered techs, especially when theres always ongoing quality
issues
>when theres so many trained certified HT looking for work?  In my
area
>of the country I can't believe how many Hospitals go this way.
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

_______________________________________________
Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

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End of Histonet Digest, Vol 63, Issue 14
****************************************