[Histonet] certification

From:"Clarke, Mary"

I have student that has completed her two years of on the job training and is now ready to get certified.  Unfortunately she has missed the cut off for taking the test, now that ASCP has changed the ways of taking the test.  Does any one know of any ways that she can get certified.  We would also be interested in any online courses.

Thanks

Terri Clarke
Histology Supervisor
All Saints Laboratory
3801 Spring Street
Racine, Wi 53405
mclarke@allsaintshealthcare.org
262-687-6609

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu
Sent: Monday, March 21, 2005 12:06 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 16, Issue 34

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Today's Topics:

   1. RE: Thin-Prep[Scanned] (Kemlo Rogerson)
   2. RE: Thin-Prep[Scanned] (Weems, Joyce)
   3. RE: AW: Automated Image analysis (Smith, Allen)
   4. RE: Industrial Sharpie good news   (Bonner, Janet)
   5. RE: .."Sharpie" marker pens (Bonner, Janet)
   6. RE: .."Sharpie" marker pens (Rebecca Barnhart)
   7. Immunofluorescence in paraffin sections
      (=?gb2312?q?=CC=EC=20=D0=C1?=)
   8. Re:Mouse histology books (Gayle Callis)
   9. NYSHS Call For Nominations 2005 (NYSHS Members Only)
      (Luis Chiriboga)
  10. RE: Immunofluorescence in paraffin sections
      (Edmondson David (RBV) NHS Christie Tr)
  11. Re: Immunofluorescence in paraffin sections (Gayle Callis)
  12. Re: Are Gloves a CAP regulation? (Osborn, Sharon)


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

Message: 1
Date: Mon, 21 Mar 2005 09:01:51 -0000
From: Kemlo Rogerson 
Subject: RE: [Histonet] Thin-Prep[Scanned]
To: 'Dave Low' , "Scholz, Stephen J."
	,
	histonet@lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D05A3F0BB@bhrv-nt-11.bhrv.nwest.nhs.uk>
	
Content-Type: text/plain

I don't think you use the ThinPrep vial, I believe you use the one that is a
preservative and lyses red blood cells, is it Cytolyse? It's in the manual.
If you use the ThinPrep vial then you have problems with rbc's which become
fixed.

-----Original Message-----
From: Dave Low [mailto:lowman034@yahoo.com] 
Sent: 18 March 2005 19:55
To: Scholz, Stephen J.; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Thin-Prep[Scanned]

Stephen,

Yes, you can use the ThinPrep vial solution with your
Non-Gyns.  Here is the contact number:

Cytyc Corporation
Customer Service
85 Swanson Road
Boxborough, MA 01719
1-800-442-9892 option 5

Good Luck!

Dave Low
Malcolm Grow Med Ctr


--- "Scholz, Stephen J."
 wrote:
> Hello all;
> 
> I was instructed by a Pathologist that I work for to
> investigate using the ThinPrep technology in a
> non-gyne capacity.  Is this done?  I know that it is
> used for gyne specimens but I have never heard of
> using the technology for non-gyne.  I'm a Histo-tech
> and my experience with cytology is limited so any
> information that I could get would be appreciated. 
> (where to get the instrument, cost, is it
> applicable)
> 
> Thank you,
> 
> Stephen J. Scholz HT(ASCP)
> 
> 815-395-5410
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 


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

Message: 2
Date: Mon, 21 Mar 2005 08:48:12 -0500
From: "Weems, Joyce" 
Subject: RE: [Histonet] Thin-Prep[Scanned]
To: "Kemlo Rogerson" ,	"Dave Low"
	,	"Scholz, Stephen J."
	,
	
Message-ID:
	<83AACDB0810528418AA106F9AE9B7F7EA9D21C@sjhaexc02.sjha.org>
Content-Type: text/plain;  charset="iso-8859-1"

There are non-gyn supplies - Cytolyt is that name. j


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kemlo Rogerson
Sent: Mon 3/21/2005 4:01 AM
To: 'Dave Low'; Scholz, Stephen J.; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Thin-Prep[Scanned]
 
I don't think you use the ThinPrep vial, I believe you use the one that is a
preservative and lyses red blood cells, is it Cytolyse? It's in the manual.
If you use the ThinPrep vial then you have problems with rbc's which become
fixed.

-----Original Message-----
From: Dave Low [mailto:lowman034@yahoo.com] 
Sent: 18 March 2005 19:55
To: Scholz, Stephen J.; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Thin-Prep[Scanned]

Stephen,

Yes, you can use the ThinPrep vial solution with your
Non-Gyns.  Here is the contact number:

Cytyc Corporation
Customer Service
85 Swanson Road
Boxborough, MA 01719
1-800-442-9892 option 5

Good Luck!

Dave Low
Malcolm Grow Med Ctr


--- "Scholz, Stephen J."
 wrote:
> Hello all;
> 
> I was instructed by a Pathologist that I work for to
> investigate using the ThinPrep technology in a
> non-gyne capacity.  Is this done?  I know that it is
> used for gyne specimens but I have never heard of
> using the technology for non-gyne.  I'm a Histo-tech
> and my experience with cytology is limited so any
> information that I could get would be appreciated. 
> (where to get the instrument, cost, is it
> applicable)
> 
> Thank you,
> 
> Stephen J. Scholz HT(ASCP)
> 
> 815-395-5410
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 


		
__________________________________ 
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Yahoo! Small Business - Try our new resources site!
http://smallbusiness.yahoo.com/resources/ 

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Thank you. Saint Josephs Health System, Inc.


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

Message: 3
Date: Mon, 21 Mar 2005 10:35:14 -0500
From: "Smith, Allen" 
Subject: RE: [Histonet] AW: Automated Image analysis
To: "Tony Henwood" 
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
	<4C051EAE581BB646BF53A749A73FBA2D1F3CA3@exchsrv01.barrynet.barry.edu>
Content-Type: text/plain;	charset="iso-8859-1"

When someone asks if a product exists, a discreet advertisement, such as
Christof Krug's, is an appropriate response.  In the case of a new product,
it is the only possible response. 

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 Tony Henwood
Sent: Tuesday, March 15, 2005 4:02 PM
To: 'Christof Krug'
Cc: 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] AW: Automated Image analysis


This seems to be a blatant addvertisement!
Christof this is not allowed on histonet


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318





-----Original Message-----
From: Christof Krug [mailto:Christof.Krug@t-online.de] 
Sent: Wednesday, 16 March 2005 1:06 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] AW: Automated Image analysis


Hi Yan
 
I have developed an automatic image analysis software (www.cvistec.com
 ) for complex biomedical images. It is best suited
for high throughput, high content image analysis and tissue microarrays.
Customized measurement solutions for your specific images can be add. If you
are interested, please get in touch with me.
 
Kind regards
Chistof Krug

Informatikb黵o Christof Krug 
Dominikstr.21 
D-81929 M黱chen 
Phone +49 89 99341972 
Mobil +49 170 5836734 
Fax +49 89 99341974 
Email info@cvistec.com
WWW www.cvistec.com  

 
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Barry University - Miami Shores, FL (http://www.barry.edu) 



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

Message: 4
Date: Mon, 21 Mar 2005 10:44:55 -0500
From: "Bonner, Janet" 
Subject: [Histonet] RE: Industrial Sharpie good news  
To: "'Gayle Callis'" , "Bonner, Janet"
	, Histonet@lists.utsouthwestern.edu
Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB41EB@fh2k093.fhmis.net>
Content-Type: text/plain;	charset="iso-8859-1"

Gayle,
    We purchase the industrial sharpie from Office Depot.  We only use them
on the cassettes, not slides.  We haven't had the problem of anyone grabbing
the wrong pen, but we keep them in a separate place -  pencil holder or
drawer - depending on the area.  We also use a cassette printer from TBS so
we don't have to use the pen all of the time.
     For slides we use the Statmark because the industrial sharpie has been
known to come off - for example, when we circle slides to indicate where we
put the gelblock section, and it is put through the stainer and the Statmark
pen has survived. Same story with labeling the slides - the industrial
sharpie faded.  Let me know if you need the Statmark information-   Janet

-----Original Message-----
From: Gayle Callis [mailto:gcallis@montana.edu]
Sent: Friday, March 18, 2005 3:43 PM
To: Bonner, Janet; Histonet@lists.utsouthwestern.edu
Subject: Industrial Sharpie good news 


Dear Janet,

Thank you for info about Industrial Sharpie ink staying on in 
solvents.  Where are you purchasing these special Sharpies?  Staples, or 
some other vendor?

I was worried that someone inadvertently grabs a run of the mill, everyday 
Sharpie marker used for labeling bottles, tape, and label their tissue 
cassettes or slides!  We have had this happen with disasterous results when 
a whole project's labeling is dissolved away because the unaware used the 
wrong Sharpie marker.

I plan to give the industrial Sharpies a try though.


Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)




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

Message: 5
Date: Mon, 21 Mar 2005 11:05:35 -0500
From: "Bonner, Janet" 
Subject: RE: [Histonet] .."Sharpie" marker pens
To: "'Bonnie Whitaker'" ,
	Histonet@lists.utsouthwestern.edu
Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB41EC@fh2k093.fhmis.net>
Content-Type: text/plain;	charset="iso-8859-1"

I wonder if it has something with the cassette manufacturer - maybe there is
a coating - like oil? We use Sakura/Allegiance/Cardinal (whatever they're
called now) with the industrial pen.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonnie
Whitaker
Sent: Friday, March 18, 2005 2:19 PM
To: Histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] .."Sharpie" marker pens


It didn't work for me either!

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LuAnn
Anderson
Sent: Friday, March 18, 2005 1:01 PM
To: Bonner, Janet; Histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Caveat about true "Sharpie" marker pens fortissue
cassettes Re:Colored Cassettes for multi institution study


The Sharpie "industrial" did not work in our processor.
LuAnn

At 11:37 AM 3/18/05, Bonner, Janet wrote:
>The Sharpie "industrial" survives the processing chemicals but we use 
>the Statmark pen for the slides.
>
>-----Original Message-----
>From: histonet-bounces@lists.utsouthwestern.edu
>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle 
>Callis
>Sent: Friday, March 18, 2005 10:51 AM
>To: John Robertson; Histonet@lists.utsouthwestern.edu
>Subject: [Histonet] Caveat about true "Sharpie" marker pens for tissue 
>cassettes Re:Colored Cassettes for multi institution study
>
>
>Caveat:  True Sharpie marker, per se, ink is washed off by processing 
>solvents.  Are you refering to the STATMARK PEN (StatLab) or Superfrost 
>Marker II pen that LOOK like "sharpie" markers but have special ink 
>that stays on plastic cassettes?
>
>     At 02:34 PM 3/17/2005, you wrote:
>
> >Hal- You might consider a cassette bearing a large letter for each 
> >institution. A sequentially bar coded set would permit accurate 
> >sample logging and tracking. These markings could be put on the side 
> >to permit traditional sharpie markings on the face. Alternatively the 
> >face could be marked with large letters leaving room for the sharpie.
> >
> >See a picture at
> >http://www.vialabel.com/bigletter.jpg
> >
> >Hope this helps Hal.
> >John
> >
> >
> >_______________________________________________
> >Histonet mailing list
> >Histonet@lists.utsouthwestern.edu
> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>
>
>_______________________________________________
>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


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

Message: 6
Date: Mon, 21 Mar 2005 11:14:38 -0500
From: "Rebecca Barnhart" 
Subject: RE: [Histonet] .."Sharpie" marker pens
To: 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

We have been able to use a sharpie industrial pen with Sakura, Ted Pella
and SurgiPath cassettes.  We don't use it on slides because it fades
just enough to passably cause problems.    

>>> "Bonner, Janet"  3/21/2005 11:05:35 AM
>>>
I wonder if it has something with the cassette manufacturer - maybe
there is
a coating - like oil? We use Sakura/Allegiance/Cardinal (whatever
they're
called now) with the industrial pen.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu 
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonnie
Whitaker
Sent: Friday, March 18, 2005 2:19 PM
To: Histonet@lists.utsouthwestern.edu 
Subject: RE: [Histonet] .."Sharpie" marker pens


It didn't work for me either!

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu 
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LuAnn
Anderson
Sent: Friday, March 18, 2005 1:01 PM
To: Bonner, Janet; Histonet@lists.utsouthwestern.edu 
Subject: RE: [Histonet] Caveat about true "Sharpie" marker pens
fortissue
cassettes Re:Colored Cassettes for multi institution study


The Sharpie "industrial" did not work in our processor.
LuAnn

At 11:37 AM 3/18/05, Bonner, Janet wrote:
>The Sharpie "industrial" survives the processing chemicals but we use

>the Statmark pen for the slides.
>
>-----Original Message-----
>From: histonet-bounces@lists.utsouthwestern.edu 
>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle 
>Callis
>Sent: Friday, March 18, 2005 10:51 AM
>To: John Robertson; Histonet@lists.utsouthwestern.edu 
>Subject: [Histonet] Caveat about true "Sharpie" marker pens for tissue

>cassettes Re:Colored Cassettes for multi institution study
>
>
>Caveat:  True Sharpie marker, per se, ink is washed off by processing

>solvents.  Are you refering to the STATMARK PEN (StatLab) or
Superfrost 
>Marker II pen that LOOK like "sharpie" markers but have special ink 
>that stays on plastic cassettes?
>
>     At 02:34 PM 3/17/2005, you wrote:
>
> >Hal- You might consider a cassette bearing a large letter for each 
> >institution. A sequentially bar coded set would permit accurate 
> >sample logging and tracking. These markings could be put on the side

> >to permit traditional sharpie markings on the face. Alternatively
the 
> >face could be marked with large letters leaving room for the
sharpie.
> >
> >See a picture at
> >http://www.vialabel.com/bigletter.jpg 
> >
> >Hope this helps Hal.
> >John
> >
> >
> >_______________________________________________
> >Histonet mailing list
> >Histonet@lists.utsouthwestern.edu 
> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>
>
>_______________________________________________
>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 


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http://lists.utsouthwestern.edu/mailman/listinfo/histonet 


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http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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

Message: 7
Date: Tue, 22 Mar 2005 00:21:02 +0800 (CST)
From: =?gb2312?q?=CC=EC=20=D0=C1?= 
Subject: [Histonet] Immunofluorescence in paraffin sections
To: histonet@lists.utsouthwestern.edu
Message-ID: <20050321162103.66846.qmail@web15504.mail.cnb.yahoo.com>
Content-Type: text/plain; charset=gb2312

Hello, all,
 
I am doing immunofluorescence in paraffin sections. I have some difficulties in it:
1: For pretreatment of tissue sections, I plan to incubate sections with trypsin solution, but I do not know :what kind of concentration should I choose ? (0.01% trypsin, 0.05% or o.1%)how long does it should last? (10min,20min,30 min)
2: Recently, I read a protocol about double immunofluorescence, it shows :Antibodies derived from different animal can be mixed and incubated as a cocktail (example: rabbit anti-A and mouse anti-B). The same is valid for secondary antibodies (example: goat anti-rabbit Texas Red conjugated and goat anti-mouse fluorescein conjugated). but If secondary antibodies cross-react, what should I do? My friend told me that I could use depletion method. but I am not sure.
 
Can anybody do me a favor?
Thank you!
 
Guofeng
 



---------------------------------
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注册世界一流品质的雅虎免=D1电邮

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

Message: 8
Date: Mon, 21 Mar 2005 09:50:33 -0700
From: Gayle Callis 
Subject: [Histonet] Re:Mouse histology books
To: Timothy Macatee ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.1.20050321093939.01b66d38@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Tim,

The only mouse histology book I know of that has color photos is 
Histological Atlas of the Laboratory Mouse, Gude, Cosgrove and 
Hirsch.  1982,  ISBN 0-306-40686-1, and probably long out of print (and 
should be returned to the fold!)  The photos are marginal at times (low 
power) or not the best quality,  but it is better than nothing at all.

There is another atlas, A colour atlas of Anatomy of Small Laboratory 
Animals, Volume 2: rat, Mouse Hamster Popesko, Rajtova and Horak from 
SaundersISBN 0 7020 2703 0, Reprint 2002.  This is pricey and will NOT have 
photos for what you are looking for - everything is in diagram form and 
more for locating organs, structures, etc.

There are websites with some photographs but you have to do some searching 
on line.

Otherwise, there really is not much out there in terms of this species nor 
even a rat.

Good luck on finding the first book.


Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 9
Date: Mon, 21 Mar 2005 10:45:05 -0500
From: Luis Chiriboga 
Subject: [Histonet] NYSHS Call For Nominations 2005 (NYSHS Members
	Only)
To: Histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain;	charset="iso-8859-1"

TO: All New York State Histotechnology Society Members
RE: Award nominations for 2005



The NYSHS Awards committee is asking for nominations from its membership for
this years awards cycle. In addition to recognition by the society,
recipients will also be awarded a small financial grant to further their
knowledge and education in the discipline of histotechnology. Applications
are due by April 10th 2004



1.Thermo Electron Student Scholarship Award:

Is awarded to a histology student or a histotech who wishes to attend a
professional meeting. This award is sponsored by thermo Electron inc. and
must be used to defray educational expenses.Please send us:

  1.. A letter from you, showing evidence of your commitment to continuing
education.
  2.. Two letters of recommendation from supervisor, pathologists, or
histotechnologist
  3.. Name and address of your current employer or school, and your current
address
2. Dominic Europa Award:

Awarded to a long standing NYSHS member (for use towards an educational
meeting) who serves as an inspiration to others in the field.  Candidate can
be a bench tech or a supervisor (preferably not in the limelight). Please
submit a nomination and recommendation letter, detailing the nominees
contributions .



3.Biogenex Excellence in Education Award:

Awarded to any member in good standing, to be used to fund an educational
endeavor.  This endeavor could be tuition for a class, educational materials
or payment for a meeting that is not funded by an employer.  To apply for
this award, please send a letter outlining how you would benefit
educationally from this award, and how it will help you to better serve the
profession.  This award is sponsored by Biogenex.



We encourage e-mail submission of applications and letters. Please submit
applications to:

Cindy Kosuda
4997 Henderson Street
Whitesboro, NY 13492
Fax (315)624-4919
cindyk@centrexlabs.com


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

Message: 10
Date: Mon, 21 Mar 2005 17:13:27 -0000
From: "Edmondson David \(RBV\) NHS Christie Tr"
	
Subject: RE: [Histonet] Immunofluorescence in paraffin sections
To: "? ?" 
Cc: "Histonet \(E-mail 2\)" 
Message-ID:
	
Content-Type: text/plain;	charset="gb2312"

Hello,
1.  I have no experience of using fluoresence BUT the pretreatments that you use to reveal antigens should be analagous to other detection systems.   SINCE IT IS THE ANTIGENS THAT ARE IMPORTANT.
So IF trypsin is the business then around 0.05% in 0.1% Calcium Chloride, for say ten minutes woks for various antigens by immunoperoxidase methods.     If the fluoresence is a sensitive as one imagines then times and concentrations may be shorter.
   The time that is best is what you find works best rather than what I say.  What about other pretreatments? 
2. Can not help here
Dave
Histology
Christie Hospital
Manchester UK

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of pex0220@yahoo.com.cn
Sent: 21 March 2005 16:21
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Immunofluorescence in paraffin sections


Hello, all,
 
I am doing immunofluorescence in paraffin sections. I have some difficulties in it:
1: For pretreatment of tissue sections, I plan to incubate sections with trypsin solution, but I do not know :what kind of concentration should I choose ? (0.01% trypsin, 0.05% or o.1%)how long does it should last? (10min,20min,30 min)
2: Recently, I read a protocol about double immunofluorescence, it shows :Antibodies derived from different animal can be mixed and incubated as a cocktail (example: rabbit anti-A and mouse anti-B). The same is valid for secondary antibodies (example: goat anti-rabbit Texas Red conjugated and goat anti-mouse fluorescein conjugated). but If secondary antibodies cross-react, what should I do? My friend told me that I could use depletion method. but I am not sure.
 
Can anybody do me a favor?
Thank you!
 
Guofeng
 



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Message: 11
Date: Mon, 21 Mar 2005 10:18:01 -0700
From: Gayle Callis 
Subject: Re: [Histonet] Immunofluorescence in paraffin sections
To: =?gb2312?q?=CC=EC=20=D0=C1?= ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.1.20050321101213.01b0cb50@gemini.msu.montana.edu>
Content-Type: text/plain; charset="iso-8859-1"; format=flowed

Goat is not the only host for seconday antibodies.  You could use Donkey 
antimouse - FITC.   Try Jackson Immunoreasearch for other antibodies that 
will NOT cross react. Jackson has a website.


At 09:21 AM 3/21/2005, you wrote:
>Hello, all,
>
>I am doing immunofluorescence in paraffin sections. I have some 
>difficulties in it:
>1: For pretreatment of tissue sections, I plan to incubate sections with 
>trypsin solution, but I do not know :what kind of concentration should I 
>choose ? (0.01% trypsin, 0.05% or o.1%)how long does it should last? 
>(10min,20min,30 min)
>2: Recently, I read a protocol about double immunofluorescence, it shows 
>:Antibodies derived from different animal can be mixed and incubated as a 
>cocktail (example: rabbit anti-A and mouse anti-B). The same is valid for 
>secondary antibodies (example: goat anti-rabbit Texas Red conjugated and 
>goat anti-mouse fluorescein conjugated). but If secondary antibodies 
>cross-react, what should I do? My friend told me that I could use 
>depletion method. but I am not sure.
>
>Can anybody do me a favor?
>Thank you!
>
>Guofeng
>
>
>
>
>---------------------------------
>Do You Yahoo!?
>注册世界一流品质的雅虎免=B7训缬
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 12
Date: Mon, 21 Mar 2005 12:42:40 -0500
From: "Osborn, Sharon" 
Subject: [Histonet] Re: Are Gloves a CAP regulation?
To: "'histonet@lists.utsouthwestern.edu'"
	
Message-ID:
	<29B25753F6B1D51196110002A589D44402397F1D@PALMSG30.us.schp.com>
Content-Type: text/plain

	As many of you, I have worked several lifetimes in histology
including using bare hands to lift specimens out of formalin.  However, I
began wearing gloves in the early 1970's because I developed sensitivity to
the formalin--fingers being numb, stinging sensations, etc.  In these later
years, I notice sensitivity to xylene--yes, it is slow going doing
coverslipping with gloves when need to do hand 'slipping.    I wear gloves
for embedding because there is still some carryover of the xylene in the
paraffins, even in the #3 paraffin.  As we have become more educated about
our hazardous, we know that xylene travels through our fatty tissue and
lodges in the liver where it remains..  Nitrile gloves also resist xylene
longer than latex.  I don't wear gloves when microtoming paraffin sections
unless it is for RNAse studies--then it is a requirement to keep my RNAse
from contaminating the study material.  I do wear gloves while using the
cryostat.
	I had not noticed anyone mentioning sensitivity problems to the
chemicals or the possible health hazards to us with absorbing the chemicals
through our skin.  We have plenty of people telling us about the smell of
chemicals when they come by and that tends to be addressed more quickly. =20

Sharon Osborn
DNAX
Palo Alto, CA


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