[Histonet] CBG recycler

From:"Gus Mondragon"


Hi Brandi; The CBG recycler is an excellent unit but you must understand a few things:
It is not very fast and if your solvents are oversaturated it will take much longer. I run my basically 24 hrs to get about 12-15 gals of alcohol or xylene, that's about 3 runs
It is very self contained, easy to operate with very little maintanance.
Your lab staff must be very careful segregating the alcohols and xylenes in order to get good results. If your xylene has any traces of water from alcohol carry over the machine will not eliminate all of this water and your recyle product will turn milky when used.
Always use a hydrometer to check your 100% which will be about 98%. We use some of this 98% to make 95% we just add water until the hydrometer reads 95% 
Use the mathematical formula in the CBG directions to check the Xylene purity, important.
Once again, segregation, segregation of solvents it the only way the recovery is going to work x you.    Gustave Mondragon Greensboro NC
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu
Sent: Wednesday, March 08, 2006 10:41 AM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 28, Issue 12


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Today's Topics:

   1. Re: Histowrap (Bill Blank)
   2. FW: Needing information about recycling (Joyce Cline)
   3. CD69, CD94 and Bax antibodies for mouse tissue
      (Elizabeth Chlipala)
   4. Hall or fouchet stain (Gudrun Lang)
   5. RE: CD69, CD94 and Bax antibodies for mouse tissue (Jacqui Detmar)
   6. Vital nerve tissue stain (in vivo) (UnJa L. Hayes)
   7. DAKO AEC+ (Richard Cartun)
   8. RE: Histonet Digest, Vol 28, Issue 11 (Ravishankar Nagarajan)
   9. Re: Vital nerve tissue stain (in vivo) (John Kiernan)
  10. Histology Managers, Supervisors,	and Bench Techs needed-
      Intervirewing and Hiring NOW (Eric Dye (ext 223))
  11. cryo soft tissue (Arvind)
  12. negative immunohistochemistry control (cynthia haynes)
  13. RE: Negative Immunohistochemistry controls (Kellar, Eric C)
  14. Re: negative immunohistochemistry control (Chris Pomajzl)
  15. Correction RE: Negative Immunohistochemistry controls
      (Kellar, Eric C)
  16. Re: negative immunohistochemistry control (Rene J Buesa)
  17. RE: negative immuno histochemistry control (Edwards, R.E.)
  18. Outsourcing transcription (Martha Ward)
  19. Re: negative immunohistochemistry control (Chris Pomajzl)
  20. RE: negative immuno histochemistry control (Dawson, Glen)
  21. RE: negative immunohistochemistry control (Sebree Linda A.)
  22. FW: STATLINE -- Special Report: CMS Announces MUE	Proposal
      Will Be Withdrawn (Weems, Joyce)
  23. Re: negative immunohistochemistry control (Jackie M O'Connor)
  24. myelin stain for 40um cryostat sections (Maria Mejia)
  25. RE: RE: Histonet Digest, Vol 28, Issue 11 (Ford Royer)
  26. RE: myelin stain for 40um cryostat sections (Kemlo Rogerson)


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

Message: 1
Date: Tue, 7 Mar 2006 12:02:47 -0600
From: Bill Blank 
Subject: [Histonet] Re: Histowrap
To: 
Message-ID: 
Content-Type: text/plain; charset="us-ascii" ; format="flowed"

We use end papers used for home permanents which are available at the 
local stores and are cheap.

BB



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

Message: 2
Date: Tue, 7 Mar 2006 13:24:15 -0500
From: "Joyce Cline" 
Subject: [Histonet] FW: Needing information about recycling
To: 
Message-ID: <003301c64214$58344ec0$1d2a14ac@wchsys.org>
Content-Type: text/plain;charset="us-ascii"

I use the CBG system. CBG should be able to answer your questions, they have been most helpful to me. I only recycle alcohol and xylene substitute.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Brandi Farris
Sent: Sunday, March 05, 2006 10:02 PM
To: histonet@lists.utsouthwestern.edu
Cc: Patricia Bates
Subject: [Histonet] Needing information about recycling

Our histology laboratory is looking at recycler from CBG that processes alcohol, xylene and formalin and we're interested in feedback and answers from users of the system. Are there any fumes with the recycler while it is processing formalin? Can we use the recycled formalin for patient specimens or is it for use only on the processor?Will you please tell us the cost and procedure for buffering? Can you estimate about how much tech time is spent with the system? Your response is appreciated.

Thank you,
Brandi Farris
Boone Hospital Center
Columbia, MO  65201



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

Message: 3
Date: Tue, 7 Mar 2006 11:27:09 -0700
From: "Elizabeth Chlipala" 
Subject: [Histonet] CD69, CD94 and Bax antibodies for mouse tissue
To: 
Message-ID: <000001c64214$bf248aa0$c6d48a80@Chlipala>
Content-Type: text/plain;	charset="us-ascii"

Hello everyone
 
I was wondering what was a good source for the following antibodies for mouse tissue.  Any suggestions would be helpful.
 
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@premierlab.com
www.premierlab.com  
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
 
 


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

Message: 4
Date: Tue, 7 Mar 2006 19:34:38 +0100
From: "Gudrun Lang" 
Subject: [Histonet] Hall or fouchet stain
To: "Histonetliste \(Histonetliste\)"
	
Message-ID: <002f01c64215$ccb52d90$eeeea8c0@SERVER01>
Content-Type: text/plain;	charset="us-ascii"

 

Can somebody help me with the shelflive of this reagens'?
25% trichloracetic acid
Fouchet reagens: 100ml 25% trichloracetic acid and 10ml 10%ferrichlorid

Thank you
Gudrun Lang

Akh Linz
Austria







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

Message: 5
Date: Tue, 7 Mar 2006 13:44:57 -0500
From: "Jacqui Detmar" 
Subject: RE: [Histonet] CD69, CD94 and Bax antibodies for mouse tissue
To: "Elizabeth Chlipala" ,
	
Message-ID:
	
Content-Type: text/plain;	charset="iso-8859-1"

I have used the BaxNT antibody from Upstate (06-499)for both IHC and Western blotting with good results...wildtype tissues (placentae, mainly) are positive and the knockout tissues are negative.  Epitomics has come out with a new rabbit monoclonal antibody for Bax which I am tempted to try, but have not yet done so.

Jacqui

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Elizabeth Chlipala
Sent: Tuesday, March 07, 2006 1:27 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] CD69, CD94 and Bax antibodies for mouse tissue


Hello everyone
 
I was wondering what was a good source for the following antibodies for mouse tissue.  Any suggestions would be helpful.
 
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@premierlab.com
www.premierlab.com  
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
 
 
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 6
Date: Tue,  7 Mar 2006 14:38:56 -0500
From: "UnJa L. Hayes" 
Subject: [Histonet] Vital nerve tissue stain (in vivo)
To: "histonet@lists.utsouthwestern.edu"
	
Message-ID: <1141760336.440de150b876b@mail-www2.oit.umass.edu>
Content-Type: text/plain; charset=ISO-8859-1


Dear Histonet,

Does anyone know of a stain I could use in vivo that would help visualize 
nerves so as to make them easier to locate?

I need to cut specific nerves in the lower abdominal area of a small 
rodent. However, I'm having difficulty finding the nerves (or all of the 
branches of the nerves). I've searched the web, but couldn't find any simple 
stains that I could use in vivo .

Any assistance would be greatly appreciated!!!!

u


-- 
UnJa L. Hayes, Ph.D.
Center for Neuroendocrine Studies
University of Massachusetts
Department of Psychology
Tobin Hall
Amherst, MA 01003
Off: (413) 545-5955
Lab: (413) 545-0526
Fax: (413) 545-0996






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

Message: 7
Date: Tue, 07 Mar 2006 18:08:03 -0500
From: "Richard Cartun" 
Subject: [Histonet] DAKO AEC+
To: 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

Is anyone having problems with DAKO's AEC+ chromogen (tiny dots everywhere).  Thanks!

Richard

Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax


                                        




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

Message: 8
Date: Wed, 8 Mar 2006 09:00:09 +0530
From: "Ravishankar Nagarajan" 
Subject: [Histonet] RE: Histonet Digest, Vol 28, Issue 11
To: 
Message-ID:
	<2533F91DF0FC3F48AE4919B3197765ED2E9619@GUR-PR-EXA1.Ranbaxy.com>
Content-Type: text/plain;	charset="iso-8859-1"




Dear Group,
  Can anybody provide me the procedure for Wright Giemsa staining procedure for Bone marrow smears. We are performing the Wright's procedure here and we get inconsistent results here. Like the nuclei do not get stained. Could it be due to the fact that Wright's is only a cytoplasmic stain. Any suggestions would be welcome. I also include the procedure by which we do Wright's here.

Wright's stain - 8 - 10 minutes
Sorensen's buffer  (pH - 7) - 8 -10 minutes

Regards and Thanks,
Dr.N>Ravishankar

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

Message: 9
Date: Wed, 08 Mar 2006 01:53:35 -0500
From: John Kiernan 
Subject: Re: [Histonet] Vital nerve tissue stain (in vivo)
To: "UnJa L. Hayes" 
Cc: "histonet@lists.utsouthwestern.edu"
	
Message-ID: <440E7F6F.B2E54CED@uwo.ca>
Content-Type: text/plain; charset=us-ascii

Yes. Methylene blue. There is a large body of
literature dating from Paul Ehrlich (1880s) to the
1990s and perhaps later. Vital staining with
methylene blue should reveal the nerves in a
surgical setting, but you will need to dig in the
literature for practical details. This is a method
that needs intelligence rather than a "protocol."
For my humble contribution see Histochemistry 40:
51-57 (1974). 

John Kiernan
London, Canada
-----------------------------
"UnJa L. Hayes" wrote:
> 
> Dear Histonet,
> 
> Does anyone know of a stain I could use in vivo that would help 
> visualize nerves so as to make them easier to locate?
> 
> I need to cut specific nerves in the lower abdominal area of a small 
> rodent. However, I'm having difficulty finding the nerves (or all of 
> the branches of the nerves). I've searched the web, but couldn't find 
> any simple stains that I could use in vivo .
> 
> Any assistance would be greatly appreciated!!!!
> 
> u
> 
> --
> UnJa L. Hayes, Ph.D.
> Center for Neuroendocrine Studies
> University of Massachusetts
> Department of Psychology
> Tobin Hall
> Amherst, MA 01003
> Off: (413) 545-5955
> Lab: (413) 545-0526
> Fax: (413) 545-0996
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 10
Date: Tue, 7 Mar 2006 20:49:03 -0500
From: Eric Dye (ext 223) 
Subject: [Histonet] Histology Managers, Supervisors,	and Bench Techs
	needed- Intervirewing and Hiring NOW
To: Histonetters 
Message-ID: 
Content-Type: text/plain

Fellow-Histonetters
I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well.

Here are some of my Hottest jobs:

1. Ohio (Full-time, Perm, Bench, 3rd Shift)

2. Ohio (Full-time, Perm, MANAGER)

3. Ohio (full-time, Perm,Bench, 3rd Shift)

4. Colorado (Full-Time, Perm, Bench, 3rd Shift)

5. Rhode Island (Full-time, Perm, Bench, 2nd Shift)

6. New Jersey (Full-time, Perm, Bench)

7. Massachusetts (Boston) (Part-time, Bench)

8. New York (Long Island)  (Full-time, Perm, SUPERVISOR)

9. Illinois (Full-time, Perm, SUPERVISOR)

10. Virginia(South) (Full-time, Perm, Bench)

11. Virginia (Full-time, Perm, LEAD TECH) 

12. West Virginia (Full-time, Perm, Bench)

The clients are 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 Recruiter
1-800-466-9919 ext 223

---------------------------------------------------------------
Ategra Systems Inc
Specialists in Permanent & Contract Staffing
800 466 9919 ext 223 - voice
407 671 6075 - fax
eric@ategra.com
To Learn More About Ategra:
http://www.ategra.com
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------------------------------

Message: 11
Date: Wed, 8 Mar 2006 17:28:19 +0530
From: "Arvind" 
Subject: [Histonet] cryo soft tissue
To: 
Message-ID: <000801c642a7$979bce60$aa00a8c0@nbrc.res.in>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=original

can any one provide detailed  protocol for cryosectioning of human fetus 
brain ageing 3 months prenatal to  01 day postnatal using cryotome

thanks in advance


Arvind Singh Pundir
National Brain Research Centre
Manesar
Gurgaon, HARYANA
INDIA
arvind@nbrc.res.in





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

Message: 12
Date: Wed, 8 Mar 2006 04:49:34 -0800 (PST)
From: cynthia haynes 
Subject: [Histonet] negative immunohistochemistry control
To: Histonet@lists.utsouthwestern.edu
Message-ID: <20060308124934.59087.qmail@web33011.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Good Morning everyone, I have a question about immuno
staining. I've been away from this type of staining
for a while and now I am doing them again on a regular
basis. Why do you run a negative control with each
run? Are you only suppose to put the normal serum on
negative control only? I've forgotten; would someone
please answer these questions for me. Thanks in
advance.

Cynthia Haynes H.T.



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

Message: 13
Date: Wed, 8 Mar 2006 08:23:21 -0500
From: "Kellar, Eric C" 
Subject: [Histonet] RE: Negative Immunohistochemistry controls
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<6843061CE6B98E4B96590D4F299618F801583BAF@qdcws0117.us.qdx.com>
Content-Type: text/plain; charset=iso-8859-1


Cynthia,

Negative controls are used to check the presence of non-specific staining. An adequate (-) control consists of replacing the specific primary antibody by another antibody produced in the same species but with unrelated specificity. When using polyclonal antisera, a preimmune serum may be used. With monoclonal antibodies, normal mouse serum or non-immune mouse ascites is applied. They are used to evaluate the non-specific binding of that isotope to the cells. They are applied in the highest concentration that is used for the specific antibodies. In indirect or multi-step techniques the primary antibody may also be omitted to evaluate the non-specific binding of the other reagents.

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







Good Morning everyone, I have a question about immuno
staining. I've been away from this type of staining
for a while and now I am doing them again on a regular
basis. Why do you run a negative control with each
run? Are you only suppose to put the normal serum on
negative control only? I've forgotten; would someone
please answer these questions for me. Thanks in
advance.

Cynthia Haynes H.T.


==========================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=========================3D==




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

Message: 14
Date: Wed, 8 Mar 2006 07:35:54 -0600
From: "Chris Pomajzl" 
Subject: Re: [Histonet] negative immunohistochemistry control
To: "HISTONET" ,	"cynthia haynes"
	
Message-ID: <002101c642b5$3d31b760$26fca8c0@CSP>
Content-Type: text/plain;	charset="iso-8859-1"

There are several alternatives for negative controls, but a common one is simply to use the buffer rinse in place of the primary antibody, thereby removing the first link in the chain. Another way is to keep everything the same and run the IHC stain on negative tissue (no antigen present in the tissue).

----- Original Message ----- 
From: "cynthia haynes" 
To: 
Sent: Wednesday, March 08, 2006 6:49 AM
Subject: [Histonet] negative immunohistochemistry control


> Good Morning everyone, I have a question about immuno staining. I've 
> been away from this type of staining for a while and now I am doing 
> them again on a regular basis. Why do you run a negative control with 
> each run? Are you only suppose to put the normal serum on
> negative control only? I've forgotten; would someone
> please answer these questions for me. Thanks in
> advance.
>
> Cynthia Haynes H.T.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet




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

Message: 15
Date: Wed, 8 Mar 2006 08:48:58 -0500
From: "Kellar, Eric C" 
Subject: [Histonet] Correction RE: Negative Immunohistochemistry
	controls
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<6843061CE6B98E4B96590D4F299618F801583BB0@qdcws0117.us.qdx.com>
Content-Type: text/plain; charset=iso-8859-1


Cynthia,

"They are used to evaluate the non-specific binding of that 'epitope' to the cells" I typed much too fast and neglected to proof read! Sorry about that!

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





==========================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=========================3D==




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

Message: 16
Date: Wed, 8 Mar 2006 05:51:35 -0800 (PST)
From: Rene J Buesa 
Subject: Re: [Histonet] negative immunohistochemistry control
To: cynthia haynes ,
	Histonet@lists.utsouthwestern.edu
Message-ID: <20060308135135.78870.qmail@web61222.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Cynthia:
  The idea of the negative control is to try to determine if any reaction detected in the case is due to specific or unspecific binding.
  You don't need to run a negative slide for each antibody you are going to test on the case, you only need a negative slide per case, no matter how many antibodies you run. The only requirement is that the negative slide has to come from the same tissue tested.
   
  What I used to do was to add buffer instead of the primary Ab and all the other reagents the same as in the slides run for specific Abs. You could also add non-immune globuline instead of the antibody but in this case you would need 1 negative control for each type Ab (1 negative for monoclonal Abs and 1 for polyclonal Abs).
  I hope this will help you.
  René J.

cynthia haynes  wrote:
  Good Morning everyone, I have a question about immuno staining. I've been away from this type of staining for a while and now I am doing them again on a regular basis. Why do you run a negative control with each run? Are you only suppose to put the normal serum on negative control only? I've forgotten; would someone please answer these questions for me. Thanks in advance.

Cynthia Haynes H.T.

_______________________________________________
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------------------------------

Message: 17
Date: Wed, 8 Mar 2006 14:08:29 -0000
From: "Edwards, R.E." 
Subject: RE: [Histonet] negative immuno histochemistry control
To: "cynthia haynes" ,
	
Message-ID:
	
Content-Type: text/plain;	charset="iso-8859-1"


For polyclonal antibodies use species specific(rabbit,goatetcetc) normal serum, or  pre-immune if available: for  mouse monoclonals use the  specific isotypic e.g.  IgG2a; simply omitting the  primary  antibody  or  replacing it  with  diluent  alone  is  no  control.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of cynthia haynes
Sent: 08 March 2006 12:50
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] negative immunohistochemistry control


Good Morning everyone, I have a question about immuno
staining. I've been away from this type of staining
for a while and now I am doing them again on a regular
basis. Why do you run a negative control with each
run? Are you only suppose to put the normal serum on
negative control only? I've forgotten; would someone
please answer these questions for me. Thanks in
advance.

Cynthia Haynes H.T.

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



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

Message: 18
Date: Wed, 8 Mar 2006 09:12:47 -0500
From: "Martha Ward" 
Subject: [Histonet] Outsourcing transcription
To: 
Message-ID:
	<61135F0455D33347B5AAE209B903A30412C89490@EXCHVS2.medctr.ad.wfubmc.edu>
	
Content-Type: text/plain;	charset="us-ascii"

I am posting this for our manager.  Our administration would like us to explore outsourcing our Pathology transcription and we would like to hear from anyone who has done this: vendor, cost, turnaround times.  We currently use CoPath and the vendor would have to be compatible with this.  Thanks in advance for your help.
 
Martha Ward
Wake Forest University Baptist Medical Center
Dept. of Pathology
Winston-Salem, NC 27157


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

Message: 19
Date: Wed, 8 Mar 2006 08:21:09 -0600
From: "Chris Pomajzl" 
Subject: Re: [Histonet] negative immunohistochemistry control
To: "HISTONET" ,	"Rene J Buesa"
	
Message-ID: <003b01c642bb$8c1baf60$26fca8c0@CSP>
Content-Type: text/plain;	charset="iso-8859-1"

I don't understand the idea of only one negative control per case, as opposed to each antibody. How does one evaluate non-specific staining of antibodies that do not have a negative control? What if non-specific staining is inherent to a certain antibody? For example, the new CAP question regarding staining of endogenous biotin, particularly in kidney and liver - if you do not run a negative control for that specific antibody, how do you know if there is non-specific staining?

----- Original Message ----- 
From: "Rene J Buesa" 
To: "cynthia haynes" ; 
Sent: Wednesday, March 08, 2006 7:51 AM
Subject: Re: [Histonet] negative immunohistochemistry control


> Cynthia:
>   The idea of the negative control is to try to determine if any 
> reaction
detected in the case is due to specific or unspecific binding.
>   You don't need to run a negative slide for each antibody you are 
> going
to test on the case, you only need a negative slide per case, no matter how many antibodies you run. The only requirement is that the negative slide has to come from the same tissue tested.
>
>   What I used to do was to add buffer instead of the primary Ab and 
> all
the other reagents the same as in the slides run for specific Abs. You could also add non-immune globuline instead of the antibody but in this case you would need 1 negative control for each type Ab (1 negative for monoclonal Abs and 1 for polyclonal Abs).
>   I hope this will help you.
>   René J.
>
> cynthia haynes  wrote:
>   Good Morning everyone, I have a question about immuno staining. I've 
> been away from this type of staining for a while and now I am doing 
> them again on a regular basis. Why do you run a negative control with 
> each run? Are you only suppose to put the normal serum on
> negative control only? I've forgotten; would someone
> please answer these questions for me. Thanks in
> advance.
>
> Cynthia Haynes H.T.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ---------------------------------
> Yahoo! Mail
> Bring photos to life! New PhotoMail  makes sharing a breeze. 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet




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

Message: 20
Date: Wed, 8 Mar 2006 08:24:30 -0600 
From: "Dawson, Glen" 
Subject: RE: [Histonet] negative immuno histochemistry control
To: "Edwards, R.E." , cynthia haynes
	, Histonet@lists.utsouthwestern.edu
Message-ID:
	<84A42E6F38BE8A45AAB03628C8C80D1204C2C6@dynams.dynacaremilwaukee.com>
Content-Type: text/plain; charset="iso-8859-1"

Although this is a good point, I don't think this is optimal unless the normal serum or pre-immune that is used for the negative is from the exact same animal/supernatant that the antibody is drawn from with the antibody seperated out.  Throwing in something "else" is bringing another factor into the equation.  How is a person to know that the "exceptable" negative was collected correctly or that it is not contaminated.

That being said, I myself do the above to remain CAP compliant.  I used to do the antibody diluent for whichever antibody I was using for the negative control & I still think it's the most logical thing to do.  By using diluent as the negative & keeping all other steps/reagents the same as the antibody you are running, you are ensuring that it is, indeed, something in the concentrated antibody that is causing staining not seen on the negative.  

Just My Opinion,

Glen Dawson  BS, HT & QIHC (ASCP)
IHC Manager
Milwaukee, WI

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Edwards, R.E.
Sent: Wednesday, March 08, 2006 8:08 AM
To: cynthia haynes; Histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] negative immuno histochemistry control



For polyclonal antibodies use species specific(rabbit,goatetcetc) normal serum, or  pre-immune if available: for  mouse monoclonals use the  specific isotypic e.g.  IgG2a; simply omitting the  primary  antibody  or  replacing it  with  diluent  alone  is  no  control.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of cynthia haynes
Sent: 08 March 2006 12:50
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] negative immunohistochemistry control


Good Morning everyone, I have a question about immuno
staining. I've been away from this type of staining
for a while and now I am doing them again on a regular
basis. Why do you run a negative control with each
run? Are you only suppose to put the normal serum on
negative control only? I've forgotten; would someone
please answer these questions for me. Thanks in
advance.

Cynthia Haynes H.T.

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

Message: 21
Date: Wed, 8 Mar 2006 08:30:45 -0600
From: "Sebree Linda A." 
Subject: RE: [Histonet] negative immunohistochemistry control
To: "Chris Pomajzl" ,	"HISTONET"
	,	"Rene J Buesa"
	
Message-ID:
	
Content-Type: text/plain;	charset="iso-8859-1"

CAP states that for panels, running one negative control using the harshest protocol suffices.

Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Chris Pomajzl
Sent: Wednesday, March 08, 2006 8:21 AM
To: HISTONET; Rene J Buesa
Subject: Re: [Histonet] negative immunohistochemistry control


I don't understand the idea of only one negative control per case, as opposed to each antibody. How does one evaluate non-specific staining of antibodies that do not have a negative control? What if non-specific staining is inherent to a certain antibody? For example, the new CAP question regarding staining of endogenous biotin, particularly in kidney and liver - if you do not run a negative control for that specific antibody, how do you know if there is non-specific staining?

----- Original Message ----- 
From: "Rene J Buesa" 
To: "cynthia haynes" ; 
Sent: Wednesday, March 08, 2006 7:51 AM
Subject: Re: [Histonet] negative immunohistochemistry control


> Cynthia:
>   The idea of the negative control is to try to determine if any 
> reaction
detected in the case is due to specific or unspecific binding.
>   You don't need to run a negative slide for each antibody you are 
> going
to test on the case, you only need a negative slide per case, no matter how many antibodies you run. The only requirement is that the negative slide has to come from the same tissue tested.
>
>   What I used to do was to add buffer instead of the primary Ab and 
> all
the other reagents the same as in the slides run for specific Abs. You could also add non-immune globuline instead of the antibody but in this case you would need 1 negative control for each type Ab (1 negative for monoclonal Abs and 1 for polyclonal Abs).
>   I hope this will help you.
>   René J.
>
> cynthia haynes  wrote:
>   Good Morning everyone, I have a question about immuno staining. I've 
> been away from this type of staining for a while and now I am doing 
> them again on a regular basis. Why do you run a negative control with 
> each run? Are you only suppose to put the normal serum on
> negative control only? I've forgotten; would someone
> please answer these questions for me. Thanks in
> advance.
>
> Cynthia Haynes H.T.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ---------------------------------
> Yahoo! Mail
> Bring photos to life! New PhotoMail  makes sharing a breeze. 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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

Message: 22
Date: Wed, 8 Mar 2006 09:34:41 -0500
From: "Weems, Joyce" 
Subject: [Histonet] FW: STATLINE -- Special Report: CMS Announces MUE
	Proposal Will Be Withdrawn
To: "Histonet" 
Message-ID:
	<83AACDB0810528418AA106F9AE9B7F7E01305A9B@sjhaexc02.sjha.org>
Content-Type: text/plain; charset="utf-8"

Good news for now! 
 
 
 
 STATLINE  
March 6, 2006  *  © 2006 College of American Pathologists Special Report CMS Announces MUE Proposal Will Be Withdrawn MUE Proposal no Longer Scheduled to be Implemented in July At a meeting today of the Physician Payment Advisory Commission (PPAC), CMS officials said that the current Medically Unbelievable Edits (MUE) proposal is no longer scheduled to be implemented in July as previously planned. The proposal will be revised and re-submitted for public review and comment. This announcement followed concerns raised by the Chairman of the Physician Payment Advisory Commission Dr. Ronald Castellanos who stated that the current edits could have significant negative impacts on patient care. Testifying before the Commission was Dr. William Rogers, who stated that he had learned about the problems with the current MUE proposal from the College of American Pathologists in a meeting last week. He had discussed these concerns with the MUE Project manager at CMS. The PPAC recommended "the CMS withdraw the proposal to create a list of MUEs and resubmit through the normal rulemaking process and to work closely with the medical community in this effort." The motion was unanimously accepted. The CMS decision to withdraw the MUE proposal comes in direct response to an intense lobbying campaign by a CAP-led coalition of national and state pathology societies. The AMA, numerous medical specialty societies, and the American Clinical Laboratory Association (ACLA) also strongly advocated for the withdrawal of the MUE proposal. The College and its coalition partners have pressed CMS to withdraw the MUE proposal and have argued that the agency should be required to utilize the formal rulemaking process to develop sweeping unit of service limits on all pathology and clinical laboratory services. The College has also asked the agency to provide the rationale and methodology utilized to develop the MUE proposal. We are awaiting a written notification from the contractor withdrawing the proposal. CAP will continue to keep members informed on further details of CMS's plans in this area.
  _____  

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

Message: 23
Date: Wed, 8 Mar 2006 08:49:23 -0600
From: "Jackie M O'Connor" 
Subject: Re: [Histonet] negative immunohistochemistry control
To: "Chris Pomajzl" 
Cc: HISTONET ,
	histonet-bounces@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="iso-8859-1"

I'm not in a clinical setting - but I use peptide inhibition as a negative 
control for each antibody.
JO'C




"Chris Pomajzl" 
Sent by: histonet-bounces@lists.utsouthwestern.edu
03/08/2006 08:21 AM

 
        To:     "HISTONET" , "Rene J Buesa" 

        cc:     (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT)
        Subject:        Re: [Histonet] negative immunohistochemistry control


I don't understand the idea of only one negative control per case, as opposed to each antibody. How does one evaluate non-specific staining of antibodies that do not have a negative control? What if non-specific staining is inherent to a certain antibody? For example, the new CAP question regarding staining of endogenous biotin, particularly in kidney 
and
liver - if you do not run a negative control for that specific antibody, 
how
do you know if there is non-specific staining?

----- Original Message ----- 
From: "Rene J Buesa" 
To: "cynthia haynes" ; 
Sent: Wednesday, March 08, 2006 7:51 AM
Subject: Re: [Histonet] negative immunohistochemistry control


> Cynthia:
>   The idea of the negative control is to try to determine if any
reaction
detected in the case is due to specific or unspecific binding.
>   You don't need to run a negative slide for each antibody you are 
> going
to test on the case, you only need a negative slide per case, no matter 
how
many antibodies you run. The only requirement is that the negative slide 
has
to come from the same tissue tested.
>
>   What I used to do was to add buffer instead of the primary Ab and 
> all
the other reagents the same as in the slides run for specific Abs. You 
could
also add non-immune globuline instead of the antibody but in this case you would need 1 negative control for each type Ab (1 negative for monoclonal Abs and 1 for polyclonal Abs).
>   I hope this will help you.
>   René J.
>
> cynthia haynes  wrote:
>   Good Morning everyone, I have a question about immuno staining. I've 
> been away from this type of staining for a while and now I am doing 
> them again on a regular basis. Why do you run a negative control with 
> each run? Are you only suppose to put the normal serum on
> negative control only? I've forgotten; would someone
> please answer these questions for me. Thanks in
> advance.
>
> Cynthia Haynes H.T.
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ---------------------------------
> Yahoo! Mail
> Bring photos to life! New PhotoMail  makes sharing a breeze. 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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

Message: 24
Date: Wed, 8 Mar 2006 07:19:00 -0800
From: Maria Mejia 
Subject: [Histonet] myelin stain for 40um cryostat sections
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed

Could someone please recommend a good myelin stain protocol for 40um  
cryostat
primate brain sections (both for fixed and fresh frozen sections? 10%  
NBF has been
used on the fixed sections. I would greatly appreciate any  
information you could
provide & thank you in advance.

Yours
Maria Bartola Mejia
University California San Francisco (UCSF)
Department of Neurosurgery
San Francisco, CA 94103
Email: mbmphoto@gmail.com



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

Message: 25
Date: Wed, 8 Mar 2006 09:27:37 -0600
From: "Ford Royer" 
Subject: RE: [Histonet] RE: Histonet Digest, Vol 28, Issue 11
To: 
Message-ID: <004201c642c4$d4da7e30$6f01a80a@fords>
Content-Type: text/plain;	charset="us-ascii"

Are you hand staining or using an automated stainer?  Back when I did BM smears, we found that they had to be run through the stainer 2 or 3 times due to the thickness of the smear.  This did work well however; and we didn't have any problems with nuclear staining or "over staining".

~ Ford

Ford M. Royer, MT(ASCP)
Minnesota Medical Specialists
Minneapolis, MN

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ravishankar Nagarajan
Sent: Tuesday, March 07, 2006 9:30 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Histonet Digest, Vol 28, Issue 11




Dear Group,
  Can anybody provide me the procedure for Wright Giemsa staining procedure for Bone marrow smears. We are performing the Wright's procedure here and we get inconsistent results here. Like the nuclei do not get stained. Could it be due to the fact that Wright's is only a cytoplasmic stain. Any suggestions would be welcome. I also include the procedure by which we do Wright's here.

Wright's stain - 8 - 10 minutes
Sorensen's buffer  (pH - 7) - 8 -10 minutes

Regards and Thanks,
Dr.N>Ravishankar

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

Message: 26
Date: Wed, 8 Mar 2006 15:35:53 -0000 
From: Kemlo Rogerson 
Subject: RE: [Histonet] myelin stain for 40um cryostat sections
To: "'Maria Mejia'" ,
	histonet@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain

Myelin can be stained with Sudan 3 and iron haematoxylin.

Cut frozen (10 U) sections and chromate in 2.5% potassium dichromate for 2 to 4 days, then wash.

5ml fresh 1% aqueous haematoxylin and 5 ml 4% iron alum fresh in a covered dish. Put your frozen sections in for 45 min at 60 degrees C. Agitate gently. Wash in water.

Decolorise in 0.5% iron alum for 1 hr with agitation.

Wash in water

Treat with 1% borax/ 2.5% potassium ferricyanide solution for 10 min with agitation.

Wash in water

6mls stock Sudan 2 (CI 12140) saturated in 99% isopropanol and dilute with 4 ml water, let stand 7 to 8 min and filter. Stain sections 10 min.

wash in water

Float onto slides, drain and mount in aqueous mountant.

This is from Lillie and I used it decades ago with success; but the old ways are usually the best.

Kemlo Rogerson
Pathology Manager
Ext  3311
DD   01934 647057
Mob 07749 754194
 



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-----Original Message-----
From: Maria Mejia [mailto:mbmphoto@gmail.com] 
Sent: Wednesday, March 08, 2006 3:19 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] myelin stain for 40um cryostat sections

Could someone please recommend a good myelin stain protocol for 40um  
cryostat
primate brain sections (both for fixed and fresh frozen sections? 10%  
NBF has been
used on the fixed sections. I would greatly appreciate any  
information you could
provide & thank you in advance.

Yours
Maria Bartola Mejia
University California San Francisco (UCSF)
Department of Neurosurgery
San Francisco, CA 94103
Email: mbmphoto@gmail.com

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Histonet mailing list
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------------------------------

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End of Histonet Digest, Vol 28, Issue 12
****************************************

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