In response to the query of altering antiobdy dilution and reducing time. In
my experience, a dilution of 1:1200 at 4C overnight is equivalent to 1:1200
for 1 hour at 37C.  Increasing the concentration may create background
staining problems despite the change in temp.  I routinely use the same
dilution overnight at 4C as I do for 1 hour at 37C with the same results.
This said a dilution of 1:1200 sounds extreme to me, but if this is what the
manufacturer recommends for the method you are using (IHC not western or
ELISA) then this is a good starting point.

Tamara
Dr. Tamara Franz-Odendaal
Biology Dept., Dalhousie University
Halifax, NS, B3H 4J1
Canada

----- Original Message ----- 
From: 
To: 
Sent: Friday, March 17, 2006 2:09 PM
Subject: Histonet Digest, Vol 28, Issue 28


Send Histonet mailing list submissions to
histonet@lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
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When replying, please edit your Subject line so it is more specific
than "Re: Contents of Histonet digest..."


Today's Topics:

   1. Somewhat rhetorical question regarding antibody incubation
      time (Jackie M O'Connor)
   2. Re: Cd8 on rat tissue (Susan Q Wells)
   3. Commercial micro waves (Doug Geddes)
   4. RE: Histonet Digest, Vol 28, Issue 27 (Orr, Rebecca)
   5. double stain (Orr, Rebecca)
   6. Re: Commercial micro waves (Phil McArdle)
   7. Training Med Techs (Orr, Rebecca)
   8. Re: Somewhat rhetorical question regarding antibody
      incubation time (Rene J Buesa)
   9. New NY regulations (KDwyer3322@aol.com)
  10. Region IX Education Day (Mark Elliott)
  11. RE: Sudan Black Staining (Kellar, Eric C)
  12. clarifying the MT -HT training (Orr, Rebecca)
  13. RE: Gram's iodine mystery solved (Smith, Allen)
  14. An aside on viewing kidney biopsies Re: [Histonet]
      microscopes (Gayle Callis)
  15. Re: New NY regulations (Judith LaDuc)
  16. immunohistochemistry and TUNEL background staining (Jacqui Detmar)
  17. Re: Sudan Black Staining (John A. Kiernan)


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

Message: 1
Date: Fri, 17 Mar 2006 06:51:56 -0600
From: "Jackie M O'Connor" 
Subject: [Histonet] Somewhat rhetorical question regarding antibody
incubation time
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="us-ascii"

If you have an antibody that the manufacturer recommends overnight
incubation at 4C at 1:1200 dilution -
is there anything to be gained from increasing the concentration (1:600)
and incubating for one hour RT instead?



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

Message: 2
Date: Fri, 17 Mar 2006 07:53:19 -0500
From: Susan Q Wells 
Subject: Re: [Histonet] Cd8 on rat tissue
To: Patsy Ruegg 
Cc: histonet@pathology.swmed.edu
Message-ID: <441AB13F.30805@bms.com>
Content-Type: text/plain; format=flowed; charset=us-ascii

I have used mouse anti-rat from BD at .1ug/ml on frozen rat sections
with Dako's LSAB2 rat kit (standard protocol) with good results.

Patsy Ruegg wrote:

>I was wondering what is being used for CD8 labeling of rat tissue?  I
assume
>that it still has to be done on frozen tissue?  I have done a lot of rat
>anti-cd8 on mouse tissue using BD antibody but now I need to do it on rat
>tissue.
>Thanks,
>Patsy
>
>Patsy Ruegg, HT(ASCP)QIHC
>IHCtech, LLC
>Fitzsimmons BioScience Park
>12635 Montview Blvd. Suite 216
>Aurora, CO 80010
>P-720-859-4060
>F-720-859-4110
>wk email pruegg@ihctech.net
>web site www.ihctech.net 
>
>
>This email is confidential and intended solely for the use of the Person(s)
>('the intended recipient') to whom it was addressed. Any views or opinions
>presented are solely those of the author. It may contain information that
is
>privileged & confidential within the meaning of applicable law. Accordingly
>any dissemination, distribution, copying, or other use of this message, or
>any of its contents, by any person other than the intended recipient may
>constitute a breach of civil or criminal law and is strictly prohibited. If
>you are NOT the intended recipient please contact the sender and dispose of
>this e-mail as soon as possible.
>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>



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

Message: 3
Date: Fri, 17 Mar 2006 08:06:53 -0500
From: "Doug Geddes" 
Subject: [Histonet] Commercial micro waves
To: 
Message-ID: 
Content-Type: text/plain; charset="US-ASCII"

Would anyone be will to share any information good or bad about
commercial mircro waves for antigen retrieval, vendors welcomed.


Doug Geddes BSc., MLT
Dept of Pathology
London Health Sciences Centre
London, Ontario, Canada

-----------------------------------------
This information is directed in confidence solely to the person
named above and may contain confidential and/or privileged
material.  This information may not otherwise be distributed,
copied or disclosed.  If you have received this e-mail in error,
please notify the sender immediately via a return e-mail and
destroy original message.  Thank you for your cooperation.




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

Message: 4
Date: Fri, 17 Mar 2006 07:32:28 -0600
From: "Orr, Rebecca" 
Subject: [Histonet] RE: Histonet Digest, Vol 28, Issue 27
To: 
Message-ID:

Content-Type: text/plain; charset="US-ASCII"

Stacy,
I wrote a sentence describing the issues of background staining.  I then
described what detection kits we use in our lab. We use both
polymer(Dako/Biocare)  and the Ventana I view on the benchmark.  I also
added instructions to determine if Endogenous Biotin is evident
(omitting primary and secondary and just adding the label and chromogen
on one slide, just the chromogen on a second slide...)
I put something in there about following manufacturer's recommendations
on the data sheets. I addressed protein blocks and endogenous peroxidase
all in the same section and called it "Blocking Reagents".
I was just inspected in February and the inspectors and I  scrutinized
that regulation especially since it was new.  I passed so I'm thinking
what I wrote was adequate.  I'll go ahead and send you the excerpt with
my references in a separate file.  Let me know if you can't receive
files from strangers.  I can fax if you'd like
Have a great weekend
Becky

 Becky Orr CLA,HT(ASCP)
IHC Lead
Evanston Northwestern Healthcare
847-570-2771


-

Message: 6
Date: Thu, 16 Mar 2006 14:01:46 -0500
From: "Stacy McLaughlin" 
Subject: [Histonet] Endogenous Biotin and CAP regulations
To: 
Message-ID:



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


 Would anyone be willing to share how they address this issue, regarding
    CAP reg: ANP.22615 "If the laboratory uses an avidin-biotin complex
    (ABC) detection system (or a related system such as
streptavidin-biotin
    or neutravidin-biotin), is there a policy that addresses nonspecific
    false positive staining from endogenous biotin?
    We're currently using the Ventana Nexes, with biotin-streptavidin
    detection system.
    Are there specific vendors your recommend for these reagents?
    Your answers are appreciated!
    Stacy
    Stacy McLaughlin HT (ASCP)
    Lead Tech, Histology
    Coolely Dickinson Hospital



Stacy McLaughlin HT (ASCP)
Lead Tech, Histology






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

Message: 5
Date: Fri, 17 Mar 2006 08:03:28 -0600
From: "Orr, Rebecca" 
Subject: [Histonet] double stain
To: 
Message-ID:

Content-Type: text/plain; charset="US-ASCII"

Rachel,
I have encountered similar issues in my journey through doublestaining.

>From what I know of Triton X, it's mainly a surfactant.  I'm not sure
of the pH.  There are solutions on the market that most likely have some
type of surfactant in them, hence all the soapy bubbles in the rinsing.
What solutions have you tried?

What I have been successful with in this type scenario is trying a
combination of either a lighter (less harsh HEIR) and then either  a
diluted enzyme or less time in the enzyme.  Can you alter the
concentration of the antibodies?  One using the triton might need higher
concentration, or more time.
When you say sequentially, are you taking one ab all the way to the
chromogen then starting the second one? I would imagine this is your
protocol, but that would be good to know.

 Becky Orr CLA,HT(ASCP)
IHC Lead
Evanston Northwestern Healthcare
847-570-2771




Message: 28
Date: Fri, 17 Mar 2006 07:42:38 -0500
From: "Emerson, Rachael" 
Subject: [Histonet] Double stain ?
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="iso-8859-1"

Hello. I am attempting to do a sequential double stain with 2 different
antibodies.
Unfortunately, one requires using Triton-X 100 for antigen retrieval and
the
other requires using trypsin.
I've run trials using several different antigen retrieval methods and
these
are the only ones that work.
I'm afraid that performing 2 different retrieval steps will cause the
stain
not to work. I would really appreciate any thoughts or suggestions.
I am working with paraformaldehyde fixed mouse embryos.


Thanks
Rachael Emerson





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

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

End of Histonet Digest, Vol 28, Issue 27
****************************************





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

Message: 6
Date: Fri, 17 Mar 2006 09:10:51 -0500
From: Phil McArdle 
Subject: Re: [Histonet] Commercial micro waves
To: Doug Geddes ,
histonet@lists.utsouthwestern.edu
Message-ID: <441AC36B.70202@ebsciences.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Hi:

Check out http://www.ebsstore.com/control/category/~category_id=C

Please feel free to contact me with any questions or issues you may have.

Best regards,

Phil McArdle
Microwave Product Manager
Energy Beam Sciences, Inc.
Tel:  800.992.9037 x 341
Cell: 860.597.6796
Fax: 860.653.0422
PMcardle@ebsciences.com
www.ebsciences.com
"ADDING BRILLIANCE TO YOUR VISION"

"I hate quotations. Tell me what you know." Ralph Waldo Emerson


Doug Geddes wrote:
> Would anyone be will to share any information good or bad about
> commercial mircro waves for antigen retrieval, vendors welcomed.
>
>
> Doug Geddes BSc., MLT
> Dept of Pathology
> London Health Sciences Centre
> London, Ontario, Canada
>
> -----------------------------------------
> This information is directed in confidence solely to the person
> named above and may contain confidential and/or privileged
> material.  This information may not otherwise be distributed,
> copied or disclosed.  If you have received this e-mail in error,
> please notify the sender immediately via a return e-mail and
> destroy original message.  Thank you for your cooperation.
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet




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

Message: 7
Date: Fri, 17 Mar 2006 09:04:02 -0600
From: "Orr, Rebecca" 
Subject: [Histonet] Training Med Techs
To: 
Cc: "Delk, Linda" 
Message-ID:

Content-Type: text/plain; charset="US-ASCII"

Hello everyone.

I would appreciate any feedback from those of you who may have had to
train MT's (ASCP) to work in Histology.

They would be trained as histo techs with the intent to promote them
into  Anatomic Pathology (Histology) management positions.

Candid comments welcome, especially from MT's who now work in histology!

To me it would be like trying to train a policeman to be a fireman,
it's a career, not a job, right?



We see a HT shortage in the Chicago area, but I am unsure how to address
this.



Degreed individuals have proven critical thinking skills via a
traditional education pathway, so I see the advantages, but to ignore
very capable HT managers with proven management and organizational
skills via non traditional pathways  is becoming an issue with me.

I mean it's not like Non degreed HT's are stooopid or something.



Thank you

Becky



 Becky Orr CLA,HT(ASCP)

IHC Lead

Evanston Northwestern Healthcare

847-570-2771





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

Message: 8
Date: Fri, 17 Mar 2006 07:04:50 -0800 (PST)
From: Rene J Buesa 
Subject: Re: [Histonet] Somewhat rhetorical question regarding
antibody incubation time
To: Jackie M O'Connor ,
histonet@lists.utsouthwestern.edu
Message-ID: <20060317150450.43863.qmail@web61217.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

If it works it will expedite your TAT and I would certainly recommend that
approach, just one thing though: you cannot equate a 2:1 increased
concentration with a 16:1 reduction of incubation time + a 6:1 fold increase
of temperature.
  You should make several tests to find out the correct new dilution factor
that will allow you to change the incubation time and temperature to what
you desire.
  Hope this will help you!
  René J.

Jackie M O'Connor  wrote:
  If you have an antibody that the manufacturer recommends overnight
incubation at 4C at 1:1200 dilution -
is there anything to be gained from increasing the concentration (1:600)
and incubating for one hour RT instead?

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





---------------------------------
Yahoo! Travel
 Find  great deals to the top 10 hottest destinations!

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

Message: 9
Date: Fri, 17 Mar 2006 10:15:43 EST
From: KDwyer3322@aol.com
Subject: [Histonet] New NY regulations
To: histonet@lists.utsouthwestern.edu
Message-ID: <22b.87e55fa.314c2c9f@aol.com>
Content-Type: text/plain; charset="ISO-8859-1"

Histonetters from New York:
As stated below will the new regulations affect histotechnicians?  Any help
would be appreciated.
Thanks,
Kathy Dwyer



The New York State Office of the Professions has posted updated information
on our state licencing of clinical laboratory technologists, technicians and
Cytotechnologists as pasted below the direct link.

http://www.op.nysed.gov/clp-mar06update.htm

March 2006
The State Education Department, with the assistance of the State Board for
Clinical Laboratory Technology and interested parties are in the process of
developing draft regulations that will be necessary for the licensure of the
professionals as clinical laboratory technologists, Cytotechnologists, and
clinical
laboratory technicians. Once these draft regulations are completed, they
will
be submitted to all interested parties and published in the State Register
for comment. At the end of this process, regulations will be presented for
consideration and action by the Board of Regents. It is anticipated that
this
process will be completed by mid-summer in 2006. Applications packets for
licensure
will not be available until the regulations are enacted. Prior to the
enactment of the regulations, the Department cannot provide any specific
information
to individuals regarding licensure under the "Special Provisions" or
"grandparenting.." As soon as the regulations are enacted, this information
will be
available.

As soon as the regulations are enacted by the State Board of Regents, the
Department will publish a Guide to Practice for the professions of Clinical
Laboratory Technology, Clinical Laboratory Technician, and Cytotechnology,
as well
as an application packet for these professions. When they are available, the
applications will be published on this website and can be downloaded and
submitted to the Department. Applicants who wish to receive a hard copy of
the Guide
to Practice and the Application Packet may submit an e-mail request for the
information now, but should not expect to receive this information before
the
summer of 2006. A notice will be put on this website when the applications
are
ready to be mailed.

Clinical Laboratory Technicians, Clinical Laboratory Technologists and
Cytotechnologists who want to request to be placed on a mailing list now for
a hard
copy of the application packet and receive it in the summer of 2006 should
send an e-mail to OPFORMS@mail.nysed.gov, giving their name and address, and
specify the packet they want by:

Code 92A and profession title: Clinical Laboratory Technologist/Clinical
Laboratory Technician, or
Code 93A and profession title: Cytotechnology.

http://www.op.nysed.gov/clp-mar06update.htm
Page last updated: 03/15/2006 12:07:36





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

Message: 10
Date: Fri, 17 Mar 2006 07:20:07 -0800
From: "Mark Elliott" 
Subject: [Histonet] Region IX Education Day
To: 
Message-ID: <441A6327020000D60000464E@mail.mrl.ubc.ca>
Content-Type: text/plain; charset=US-ASCII

Hi everyone
For anyone wanting an excuse to visit the wonderful city of Montreal, we
have the perfect reason-Region IX is hosting an Education Day on May 6th,
2006 in this vibrant city.  We are having 5 speakers on a variety of topics
and 13 vendors will be present showcasing their products.  Registration
deadline is April 14th.  Please visit
http://www.nshregionix.org/education.html
for more information, including registration forms, speaker schedule and
hotel information.
Hope to see you there!
Mark Elliott
Region IX Education Committee Chair




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

Message: 11
Date: Fri, 17 Mar 2006 10:24:16 -0500
From: "Kellar, Eric C" 
Subject: [Histonet] RE: Sudan Black Staining
To: histonet@lists.utsouthwestern.edu
Message-ID:
<6843061CE6B98E4B96590D4F299618F801583BB6@qdcws0117.us.qdx.com>
Content-Type: text/plain; charset=iso-8859-1

To slow Sudan Black B (a nonionic, hydrophobic dye) 'bleeding' on unfixed
cryostat sections, try running a duplicate slide and compare with this
method:

1. Air dry duplicate slide after staining.
2. Clear slide through 3 changes of fresh Xylene or Xylene Substitute.
3. Glass coverslip using a permanent mounting media.

The synthetic mountant will eventually dissolve some of the dye-lipid
complex, but it will give you a longer window of time to microscopically
examine your slides.


Eric C. Kellar
Histology/Immunohistochemistry
Quest Diagnostics Inc. Miami


Hi,
I am working on the Sudan Black stain.  Do the slides need to be read
immediately due to bleeding of the stain after approximately 24 hours?

Thanks for any information you can give me about the stain.
Sharon Willman


==========================3D=========================3D=========================3D
==
The contents of this message, together with any attachments, are intended
only for the use of the person(s) to which they are addressed and may
contain confidential and/or privileged information. Further, any medical
information herein is confidential and protected by law. It is unlawful for
unauthorized persons to use, review, copy, disclose, or disseminate
confidential medical information. If you are not the intended recipient,
immediately advise the sender and delete this message and any attachments.
Any distribution, or copying of this message, or any attachment, is
prohibited.
==========================3D=========================3D=========================3D
==




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

Message: 12
Date: Fri, 17 Mar 2006 09:27:50 -0600
From: "Orr, Rebecca" 
Subject: [Histonet] clarifying the MT -HT training
To: 
Message-ID:

Content-Type: text/plain; charset="US-ASCII"

In clarification,

I don't think I worded that right.  It's a salary compensation issue.  A
non degreed HT  is expected to take on the same responsibilities  as a
histology supervisor for 10K less than  a trained MT crossing over to
HT.

So to reiterate, it's not so much being eligible to handle to
management, it's about taking less salary for the same responsibility.



Thank you all for the feedback this is seems like a warm issue!





 Becky Orr CLA,HT(ASCP)

IHC Lead

Evanston Northwestern Healthcare

847-570-2771





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

Message: 13
Date: Fri, 17 Mar 2006 10:35:48 -0500
From: "Smith, Allen" 
Subject: RE: [Histonet] Gram's iodine mystery solved
To: "John Kiernan" 
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
<5D2189E74151CC42BEC02906BA8996322B919F@exchsrv01.barrynet.barry.edu>
Content-Type: text/plain; charset="US-ASCII"

   Shockingly, labels are not always correct. I have a jar labelled
"indigocarmine" whose contents is insoluble in water.
   I once (20 years ago) opened a brand new bottle labelled "acetic
anhydride" and caught a whiff of isoamyl acetate.  The contents also boiled
at the boiling point of isoamyl acetate (4 C higher than acetic anhydride),
and it did not dissolve in hot water.
   I have also borrowed a bottle of bromine from a colleague (25 years ago
at another school) and found that it performed brominations fairly well, but
required an elevated temperature to do them.  I never did find out what the
contaminant was or how it got there.

Allen A. Smith, Ph.D.
Professor of Anatomy
Barry University School of Graduate Medical Sciences
    Podiatric Medicine and Surgery
Miami Shores, Florida  33161


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan
Sent: Thursday, March 16, 2006 11:43 PM
To: meint002
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Gram's iodine mystery solved

Is this the end of the matter? Who swapped your
iodine for iron filings? Do you have an enemy
doing horrid things with the chemicals on your
shelves?
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
    kiernan[AT]uwo.ca
    http://publish.uwo.ca/~jkiernan/
    http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
meint002 wrote:
> Dear Histos,
>
> Thanks to all for your advice about making Gram's Iodine soln.
>
> There was something wrong about the reagent iodine I was using.  I
borrowed
> the proper iodine from another histo lab (thanks Luann) and now all is
just
> fine.  Dissolving it in the Potassium iodide worked like a charm.
>
>
> Joyce Meints
> Histologist
>
> University of Minnesota
> Paul and Sheila Wellstone Muscular Dystrophy Center
> MMC 206
> 420 Delaware St. SE
> Minneapolis, MN 55455-0392
>
> e-mail:    meint002@umn.edu
> lab phone: 612-626-4703
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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



The information transmitted is intended only for the person or entity to
which it is addressed and may contain confidential, and/or privileged
material. No confidentiality or privilege is waived or lost by any errant
transmission. If you receive this message in error, please immediately
delete it and all copies of it from your system and notify the sender.
E-mail transmission cannot be guaranteed to be secure or error-free as
information could be intercepted, corrupted, lost, destroyed, arrive late or
incomplete, or contain viruses.
Barry University - Miami Shores, FL (http://www.barry.edu)



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

Message: 14
Date: Fri, 17 Mar 2006 09:04:50 -0700
From: Gayle Callis 
Subject: An aside on viewing kidney biopsies Re: [Histonet]
microscopes
To: "Santana, Diane" ,
Histonet@lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20060317090020.01b46438@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

We supplied laboratories with slides that have the wells in them, commonly
used for viewing larger creatures like insects, etc.  These are sold by
Fisher, etc The biospy was floated in PBS, and you can actually use any
microscope at lowest power or stereomicroscope to look for
glomeruli.   This prevented crushing  and flattening the biopsy, a problem
we encountered far to many times when bx was examined between a slide and
coverslip immediately after removal.

At 10:55 AM 3/16/2006, you wrote:
>I am looking at microscopes for kidney bx's. My dept will start assisting
>with the bx's after the bx has been removed. I have not done this in
>YEARS... then we watch the tissue, if it floated it was probably fat, if it
>sank to the bottom of the container it was more than likely tissue. So I am
>looking for a scope that I can see if I have tissue and some light reading
>as a refresher course.

Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717




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

Message: 15
Date: Fri, 17 Mar 2006 11:23:54 -0500
From: "Judith LaDuc" 
Subject: Re: [Histonet] New NY regulations
To: , 
Message-ID: <441A9AB5.2A60.00B8.0@trudeauinstitute.org>
Content-Type: text/plain; charset=US-ASCII

Hi Kathy and NY Histotechicians/technologists,

Yes, the new regulations will affect us. We are classified under the
heading of clinical laboratory technicians/technologists and are subject
to licensure.  I would recommend signing up for the mailing list and
checking into the website routinely.

The following is a post from the Histonet archives that has a few links
and a timeline of events.
http://www.histosearch.com/histonet/Jan06/HistonetNYSLicensure.html
Dr. Kathleen Doyle from NYS will be presenting an informational session
at the Region 1 meeting in Saratoga Springs on Friday, April 28th. The
Region 1 program (April 28th & 29th) can be accessed at
http://www.nyhisto.org

Judy LaDuc, BS HTL ASCP
President, NYS Histotechnological Society

>>>  03/17/06 10:15 am >>>
Histonetters from New York:
As stated below will the new regulations affect histotechnicians?  Any
help
would be appreciated.
Thanks,
Kathy Dwyer



The New York State Office of the Professions has posted updated
information
on our state licencing of clinical laboratory technologists,
technicians and
Cytotechnologists as pasted below the direct link.

http://www.op.nysed.gov/clp- mar06update.htm

March 2006
The State Education Department, with the assistance of the State Board
for
Clinical Laboratory Technology and interested parties are in the
process of
developing draft regulations that will be necessary for the licensure
of the
professionals as clinical laboratory technologists, Cytotechnologists,
and clinical
laboratory technicians. Once these draft regulations are completed,
they will
be submitted to all interested parties and published in the State
Register
for comment. At the end of this process, regulations will be presented
for
consideration and action by the Board of Regents. It is anticipated
that this
process will be completed by mid- summer in 2006. Applications packets
for licensure
will not be available until the regulations are enacted. Prior to the
enactment of the regulations, the Department cannot provide any
specific information
to individuals regarding licensure under the "Special Provisions" or
"grandparenting.." As soon as the regulations are enacted, this
information will be
available.

As soon as the regulations are enacted by the State Board of Regents,
the
Department will publish a Guide to Practice for the professions of
Clinical
Laboratory Technology, Clinical Laboratory Technician, and
Cytotechnology, as well
as an application packet for these professions. When they are
available, the
applications will be published on this website and can be downloaded
and
submitted to the Department. Applicants who wish to receive a hard copy
of the Guide
to Practice and the Application Packet may submit an e- mail request
for the
information now, but should not expect to receive this information
before the
summer of 2006. A notice will be put on this website when the
applications are
ready to be mailed.

Clinical Laboratory Technicians, Clinical Laboratory Technologists and

Cytotechnologists who want to request to be placed on a mailing list
now for a hard
copy of the application packet and receive it in the summer of 2006
should
send an e- mail to OPFORMS@mail.nysed.gov, giving their name and
address, and
specify the packet they want by:

Code 92A and profession title: Clinical Laboratory
Technologist/Clinical
Laboratory Technician, or
Code 93A and profession title: Cytotechnology.

http://www.op.nysed.gov/clp- mar06update.htm
Page last updated: 03/15/2006 12:07:36



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

Message: 16
Date: Fri, 17 Mar 2006 11:34:15 -0500
From: "Jacqui Detmar" 
Subject: [Histonet] immunohistochemistry and TUNEL background staining
To: 
Message-ID:

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

Hi all.  I have been doing IHC and TUNELs on mouse placental tissues for the
past 4 years and have been having problems with oodles of background
staining during the past 6 months or so.  I have tried doing avidin-biotin
and streptavidin-biotin blocking and also using ABC (purchased from Vector)
in high pH (TBS, pH 9.4) or high salt (TBS [NaCl = 0.5M], pH 7.6) solutions.
Nothing seems to be working.  The IHC using biotinylated secondary
antibodies and the TUNEL is done using biotinylated dUTP.  I have tried
using old sections that I *know* have worked in the past and I am getting
the same type of background staining as my newer blocks (i.e. the problem is
likely not due to fixation (always the same...24 hours in 10%
phosphate-buffered formalin), nor due to differences in paraffin embedding).

So, could my xylene and ethanol solutions be a problem?  We get the ethanol
in-house here at the hospital and the Xylene is purchased through Fisher
(X16-4).  What about differences in TBS or PBS (I have tried using both to
see if the problem would go away and it doesn't make a difference).

Any advice would be appreciated.

Regards,

Jacqui Detmar, Ph.D. student
Samuel Lunenfeld Research Institute
Mount Sinai Hospital,
Toronto, Ontario, Canada


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

Message: 17
Date: Fri, 17 Mar 2006 12:03:44 -0500
From: "John A. Kiernan" 
Subject: Re: [Histonet] Sudan Black Staining
To: Sharon E Willman 
Cc: histonet@lists.utsouthwestern.edu
Message-ID: <441AEBF0.58CE6C9C@uwo.ca>
Content-Type: text/plain; charset=us-ascii

Sudan black B shouldn't bleed. After staining,
rinse in 70% alcohol until non-lipid structures
are destained, wash in water, and mount in an
aqueous medium. The slides are good for months -
probably longer.
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
Sharon E Willman wrote:
>
> Hi,
> I am working on the Sudan Black stain.  Do the slides need to be read
> immediately due to bleeding of the stain after approximately 24 hours?
>
> Thanks for any information you can give me about the stain.
> Sharon Willman
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

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End of Histonet Digest, Vol 28, Issue 28
****************************************



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