[Histonet] negative controls

Fri Aug 5 13:25:32 CDT 2005

Amy,
are you running commercial or novel antibodies? You're secondary may not be clean also. I've had problems with a dirty secondary causing positive staining, I got a different secondary and it was fine. To have a true negative control, you do need to run the same protocol as the positive or the tissue isn't going through the same solutions and can't be considered a true negative.
Yvonne

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Today's Topics:

1. cytospin preps and plus slides (Elizabeth Chlipala)
2. Casp-3 advice (9msm8 <@t> qlink.queensu.ca)
3. RE: fixation of fresh frozen section (Lewis, Sarah)
4. PLP fixation info RE: fixation of fresh frozen section
(Gayle Callis)
5. Re: Re: Disposal of glass slides (cindipqr <@t> wowway.com)
6. Histo Tech Mananger Supervisor and Bench Tech's needed for
immediate openings (Eric Dye (ext 223))
7. Histo Tech Mananger Supervisor, Bench Tech's and Temps needed
for immediate openings (this is not a duplicate) (Eric Dye (ext 223))
8. Problems with certification (Meryl Roberts)
9. RE: Re: ooops, rat on rat (Mandy Townsend)
10. RE: Re: Disposal of glass slides[Scanned]
(Rogerson Kemlo (ELHT) Pathology)
11. RE: slide disposal[Scanned] (Rogerson Kemlo (ELHT) Pathology)
12. RE: Whole Slide Imaging[Scanned] (Rogerson Kemlo (ELHT) Pathology)
13. RE: Whole Slide Imaging[Scanned] (Rittman, Barry R)
14. RE: cytospin preps and plus slides (Sherri Anderson)
15. Re: Problems with certification (Larry Woody)
16. RE: Problems with certification (Rittman, Barry R)
17. RE: cytospin preps and plus slides
(Rogerson Kemlo (ELHT) Pathology)
18. Negative controls (Amy Johnson)
19. Slide trays -wooden (Steven P Postl)
20. basic auto stainer for paps (Tom McNemar)
21. Tissues for HTL Practical exam needed (tracy.bergeron <@t> crl.com)
22. Histo Tech Mananger Supervisor and Bench Tech's needed for
immediate openings (Eric Dye (ext 223))
23. VEGF-R-2 (Vernon Dailey)

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

Message: 1
Date: Wed, 3 Aug 2005 12:42:30 -0600
From: "Elizabeth Chlipala" 

Subject: [Histonet] cytospin preps and plus slides
To: "'Histonet'" 
Message-ID: <000001c5985b$1c3023d0$a7d48a80 <@t> AMY>
Content-Type: text/plain; charset="us-ascii"

Hello Everyone

I'm working on a project for a client who is trying to isolate prostate
epithelial cells. I'm getting the cells submitted to me fixed in 10%
NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm
and plus slides. The problem is that I'm not getting the correct yield
of cells. They check the number of cells in the prep prior to me
receiving them. They are submitting numbers of cells in the thousands,
but when I prepare the cytospin slides, I'm only getting cells numbering
in the hundreds and sometimes less. Does anyone have any suggestions?

Thanks in advance

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com

Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

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

Message: 2
Date: Wed, 3 Aug 2005 14:53:35 -0400 (EDT)
From: 9msm8 <@t> qlink.queensu.ca
Subject: [Histonet] Casp-3 advice
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <51618.192.75.165.28.1123095215.squirrel <@t> qlink.queensu.ca>
Content-Type: text/plain;charset=iso-8859-1

Hello Histonet.
I am a student in a lab where histologists are rare to non-existant. I am
working with 5 micrometer thick mouse hippocampal parrafin embedded
sections (I used paraformaldehyde) and I want to do an
immunohistochemical stain for casp-3. I am using the marker as a detection
for relatively early apoptosis. The QUESTION I am asking is: Should I be
using this method with a fluorescence signal or a horshradish peroxidase
counterstain? Given that I probably will want my analysis to be more of a
qualitative descritpion. What should I be aware of before making my
decision? What are the advantages and disadvantages?
Any help will be greatly appreciated.

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

Message: 3
Date: Wed, 3 Aug 2005 15:32:00 -0400
From: "Lewis, Sarah" 
Subject: RE: [Histonet] fixation of fresh frozen section
To: "Edmondson David \(RBV\) NHS Christie Tr"
, "HISTONET \(E-mail\)"

Message-ID:
<714B9F12B4E18C4C843B66E8E190F2AD447562 <@t> res2k3ms01.CRII.ORG>
Content-Type: text/plain; charset="iso-8859-1"

I would love more information on this subject. Thanks in advance!! 
Sarah E. Lewis 
Histotechnician
Childrens Research
Center for Gene Therapy 
700 Childrens Dr Rm WA3112
Columbus OH 43205
(614)-722-2204
LewisS <@t> ccri.net

-----Original Message-----
From: Edmondson David (RBV) NHS Christie Tr
[mailto:David.Edmondson <@t> christie-tr.nwest.nhs.uk]
Sent: Wednesday, August 03, 2005 12:37 PM
To: Bruijntjes, J.P.
Cc: Histonet (E-mail 2)
Subject: RE: [Histonet] fixation of fresh frozen section

Hi
I recently heard a presentation where they spoke of Periodate-Lysine-Paraformaldehyde in the context of Frozen sections. The coagulant fixatives do not alter the proteins chemistry so on might suggest no loss of antigenicity, the "Gold Standard". But the morphology of PLP is much better. It is fifteen and more years since I used it myself and maybe a search would be a quicker way to find the concentrations. I ahve an email contact if you need to ask him.
David
Christie
Manchester UK
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of
Bruijntjes, J.P.
Sent: 03 August 2005 14:39
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] fixation of fresh frozen section

Hi all

When I do immunocytochemistry on fresh frozen section I fix most, if not
all slides with acetone. Sometimes I use cold methanol because the
datasheet tells me to do so. Is anyone of you who prefer other
fixatives?

Joost Bruijntjes

TNO Quality of Life

Zeist

Holland

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

Message: 4
Date: Wed, 03 Aug 2005 14:55:57 -0600
From: Gayle Callis 
Subject: [Histonet] PLP fixation info RE: fixation of fresh frozen
section
To: "Lewis, Sarah" ,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20050803134024.01b5c108 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Sarah,

This was a previous message on Histonet concerning PLP fixation. We have 
great success with this for mouse and hamster perfusion fixation for a 
prion study then paraffin processing of tissues. The recipe for PLP has 
been put on Histonet previously, going to Histonet archives should access 
fixative recipe. I haven't had success with frozen section fixation, 
although some have done so, my impatience in taking time to work with it to 
do immunofluorescence work.

Bob Chiovette wrote:
The recipe for PLP (Periodate-Lysine-Paraformaldehyde) fixative and a 
sample protocol (at least for immuno) using perfusion fixation is on the 
Chemicon website at: 
http://www.chemicon.com/techsupp/Protocol/perfusion.asp 

There are a few instances of using PLP with paraffin embedding, so the 
processing should work OK. Whether you can retrieve reactivity from 
paraffin sections seems to be related to the antigen. For example, see the 
NIEHS website:
http://dir.niehs.nih.gov/dirlep/immuno/protocols.htm 

Good luck

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: Wed, 3 Aug 2005 18:17:07 -0600
From: cindipqr <@t> wowway.com
Subject: Re: [Histonet] Re: Disposal of glass slides
To: Gayle Callis , "Clarke, Mary"
, Histonet <@t> lists.utsouthwestern.edu
Cc: histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID: <20050803231204.M24570 <@t> wowway.com>
Content-Type: text/plain; charset=iso-8859-1

Our facility likes them in smaller boxes, taped shut and labeled as glass 
slides for disposal. This way it's safe and not as heavy and bulky as the 
regular glass disposal boxes.
Cindi
HFH
Detroit Mi

---------- Original Message -----------
From: Gayle Callis 
To: "Clarke, Mary" , 
Histonet <@t> lists.utsouthwestern.edu
Sent: Wed, 03 Aug 2005 10:30:38 -0600
Subject: [Histonet] Re: Disposal of glass slides

> If you can dispose of slides in trash, how about large broken glass 
> diposal box - safer for janitors and other personnel.
> 
> At 09:20 AM 8/3/2005, you wrote:
> >My safety officer has told us that we can dispose of glass slides in the
> >regular trash, because the tissue is fixed and considered no longer
> >hazardous.
> >
> >Terri Clarke
> 
> 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 <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------- End of Original Message -------

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

Message: 6
Date: Wed, 3 Aug 2005 11:43:28 -0400
From: Eric Dye (ext 223) 
Subject: [Histonet] Histo Tech Mananger Supervisor and Bench Tech's
needed for immediate openings
To: Histonetters 
Message-ID: 
Content-Type: text/plain

Histonetters
I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions.

Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor.
I have positions in California and Michigan who are seeking Histotech Supervisors.

I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts.

The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223

Thank you,
Eric Dye-Sr Allied Healthcare Recriuter
1-800-466-9919 ext 223

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

Message: 7
Date: Wed, 3 Aug 2005 21:42:59 -0400
From: Eric Dye (ext 223) 
Subject: [Histonet] Histo Tech Mananger Supervisor, Bench Tech's and
Temps needed for immediate openings (this is not a duplicate)
To: Histonetters 
Message-ID: 
Content-Type: text/plain

Histonetters
I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions.

Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor.
I have positions in California and Michigan who are seeking Histotech Supervisors.

I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts.

I also have a temp position available in South Carolina at a Dermapathology lab. The assignment would be for 1 to 2 months and you must have dermpath experience..
This job has a deadline of 8/12/05. Please call ASAP if your interested....
There is also an opening for a Histotech traverler in Southern Florida.

The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223

Thank you,
Eric Dye-Sr Allied Healthcare Recriuter
1-800-466-9919 ext 223

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

Message: 8
Date: Thu, 04 Aug 2005 08:31:57 +0100
From: "Meryl Roberts" 
Subject: [Histonet] Problems with certification
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; format=flowed

Hi there,

I am a histologist in the UK with nearly 7 years experience, but I wish to 
move to the USA. However I'm having terrible problems finding work as it 
seems it's virtually impossible for me to become certified in the USA as 
none of my work experience is in an American lab- I've spoken to the ASCP 
and the NCA and they both say that I'm not elligeable to apply, and I'm 
currently waiting to hear back from the AMT (fingers crossed!).....is there 
any way I can become certified or should I just give up? I'm starting to get 
a bit despondant.

sincerely, Meryl Roberts.

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

Message: 9
Date: Thu, 4 Aug 2005 08:50:35 +0100
From: "Mandy Townsend" 
Subject: RE: [Histonet] Re: ooops, rat on rat
To: "Gayle Callis" , "Till, Renee"
, 
Message-ID:

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

Serotec are able to supply a polymer based kit that is suitable for use in rat on rat (and mouse on mouse) protocols - STAR4000A. Please contact our US office for further information.

Serotec North & South America (website)
3200 Atlantic Avenue, Suite 105
Raleigh, NC 27604, USA
Toll free: 1-800-265-7376 
Fax: 919-878-3751
serotec <@t> serotec-inc.com 
Product enquiry

I hope this helps.

Mandy Townsend MSc
Technical Services Advisor
Serotec Ltd
22 Bankside
Station Approach
Kidlington
Oxfordshire
OX5 1JE 

Tel: +44 1865 852733
Fax: +44 1865 852739
email: mandy <@t> serotec.co.uk
URL: http://www.serotec.com/
Serotec-Your first choice for antibodies! 

Join our free email update service 

IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible.

Serotec Ltd. Registered in England No.1604642
Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Gayle Callis
Sent: Tuesday, August 02, 2005 9:21 PM
To: Till, Renee; Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: ooops, rat on rat

You could use a biotinylated primary, as in the DAKO ARK methodology,
eliminate the secondary to avoid having some "host" antiRat detect not only
the rat primary, but also bind to rat tissue, then come back with a
Strepavidin-HRP or AP. Sometimes you have to dilute out the primary more -

I am not sure if anyone has a rat on rat kit, try Scytek out of Logan UT,
you might get lucky on a kit.

At 01:49 PM 8/2/2005, you wrote:
>Okay, I knew I'd say that wrong. It is a rat anti-mouse antibody to be
>used on rat tissues. How would you handle that?
>
>
>
>Renee' Till, HT
>
>
>
>Arkansas Childrens Nutrition Center
>
>1120 Marshall
>
>Little Rock, AR
>
>72202
>
>
>
>(501) 364-2774
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> 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)

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

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

Message: 10
Date: Thu, 4 Aug 2005 08:13:14 +0100
From: "Rogerson Kemlo \(ELHT\) Pathology" 
Subject: RE: [Histonet] Re: Disposal of glass slides[Scanned]
To: "Gayle Callis" , "Clarke, Mary"
, 
Message-ID:
<53A22BBB01D6184EBF7A4DC18A021E9E698E53 <@t> elht-exch1.xelht.nhs.uk>
Content-Type: text/plain; charset="us-ascii"

But what about confidentiality? You've got to grind them up first!

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gayle
Callis
Sent: 03 August 2005 17:31
To: Clarke, Mary; Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Disposal of glass slides[Scanned]

If you can dispose of slides in trash, how about large broken glass
diposal 
box - safer for janitors and other personnel.

At 09:20 AM 8/3/2005, you wrote:
>My safety officer has told us that we can dispose of glass slides in
the
>regular trash, because the tissue is fixed and considered no longer
>hazardous.
>
>Terri Clarke

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

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

Message: 11
Date: Thu, 4 Aug 2005 08:07:04 +0100
From: "Rogerson Kemlo \(ELHT\) Pathology" 
Subject: RE: [Histonet] slide disposal[Scanned]
To: "Bonnie Whitaker" ,

Message-ID:
<53A22BBB01D6184EBF7A4DC18A021E9E698E50 <@t> elht-exch1.xelht.nhs.uk>
Content-Type: text/plain; charset="us-ascii"

Hire a cement mixer and get some lumps of metal or ball bearings. Put
the slides and bearings in the mixer, turn on. Voila powdered glass;
take to dump.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bonnie
Whitaker
Sent: 02 August 2005 21:03
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] slide disposal[Scanned]

Hi All,

Can some of you give me ideas of how you handle old slide disposal?
Particularly those of you in smaller, private labs. I have about 20
years
worth of slides that I need to dispose of. If I have to send them with
my
biohazardous waste, I can, but it seems like an unnecessary expense.
Currently that is what the procedure says that we do, but is it really
necessary? Maybe someone out there would like to build a glass house :
) 

Bonnie Whitaker
Lab Manager
Brown & Associates Medical Laboratories
8076 El Rio
Houston, Texas 77054
713-741-6677

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

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

Message: 12
Date: Thu, 4 Aug 2005 08:09:42 +0100
From: "Rogerson Kemlo \(ELHT\) Pathology" 
Subject: RE: [Histonet] Whole Slide Imaging[Scanned]
To: "Bryan Llewellyn" , "Histonet"

Message-ID:
<53A22BBB01D6184EBF7A4DC18A021E9E698E51 <@t> elht-exch1.xelht.nhs.uk>
Content-Type: text/plain; charset="iso-8859-1"

I'm sure you are correct but the initial bargaining went wrong when we sent an upper class twit with an attitude to the negotiating table; so I'm told.

Never understood why we have less disposable earnings than the Americans AND everything costs more.

=== message truncated ===

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