[Histonet] RE: Histonet Digest, Vol 13, Issue 6

Fri Dec 3 11:12:22 CST 2004

To Darren Hess,
I used PAP pens from Vector Laboratories for years and they work great, no reaction what so ever.

Sherif Girees,B.S.HT(ASCP)QIHC
Manager of Molecular Pathology.
Pathway Diagnostics Inc.
11215 Knott Avenue. 
Cypess, Ca 90630.
Tel. 714-892-1201.
Fax 714-892-1211.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Friday, December 03, 2004 8:25 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 13, Issue 6

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Today's Topics:

   1. RE: New JCAHO PI Standard (Horn, Hazel V)
   2. immunohistochemical marker for osteoblasts (Elizabeth Chlipala)
   3. [BULK] - RE: [Histonet] static electricity (Fred Underwood)
   4. X-gal and pap pens? (hessd101 <@t> comcast.net)
   5. RE: fixation for RE: Masson's Trichrome on frozen heart
      sections (GUTIERREZ, JUAN)
   6. RE: static electricity/humidity problems (rgrow <@t> bmnet.com)
   7. Re: X-gal and pap pens? (Gayle Callis)
   8. RE: Destaining DifQuik (Tony Henwood)
   9. C4D Vendor (WWmn916 <@t> aol.com)
  10. RE: RE: static electricity/humidity problems
      (bettina.hutz <@t> orionpharma.com)
  11. RE: Destaining DifQuik[Scanned] (Kemlo Rogerson)
  12. RE: Destaining DifQuik[Scanned] (Kemlo Rogerson)
  13. RE: M.tuberculosis in formalin-fixed tissue[Scanned]
      (Kemlo Rogerson)
  14. cryosection fetal tissue (Arvind Pundir)
  15. Re: Re: Ammonia water pH and other bluing	solutions[Scanned]
      (lpwenk <@t> sbcglobal.net)
  16. RE: Re: Ammonia water pH and other bluing solutions[Sc	anned]
      (Kemlo Rogerson)
  17. RE: RE: static electricity/humidity problems (Weems, Joyce)
  18. RE: C4D Vendor (GUTIERREZ, JUAN)
  19. RE: C4D Vendor (Sebree Linda A.)
  20. RE: RE: static electricity/humidity problems (Philip Oshel)
  21. RE: RE: static electricity/humidity problems (Weems, Joyce)

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

Message: 1
Date: Thu, 2 Dec 2004 12:08:14 -0600
From: "Horn, Hazel V" <HornHV <@t> archildrens.org>
Subject: RE: [Histonet] New JCAHO PI Standard
To: "Nita Searcy" <NSEARCY <@t> swmail.sw.org>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<9AE8AA9E1F644B4AA6C155FB6FD51C6322B9BD <@t> EMAIL.archildrens.org>
Content-Type: text/plain; charset=us-ascii

Our pathologists do this.   All of our cases are assigned a code and a
report is pulled up every month using the codes for review by the
surgical affair committee.

Hazel Horn, HT/HTL (ASCP)
Histology Supervisor
Arkansas Children's Hospital

Phone - 501.364.4240
Fax - 501.364.3912 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nita
Searcy
Sent: Thursday, December 02, 2004 7:50 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] New JCAHO PI Standard

There is a new standard that states:
"An analysis is performed for all major discrepancies between
preoperative and postoperative (including pathologic) diagnoses."

How are institutions complying ? In my past lives there were "Tissue
Committees" that reviewed "normal" tissues but  this standard , in my
opinion, would not be able to be handled in the pathology lab. We
consider ourselves lucky if we get any preoperative diagnosis on our
requisitions received from surgery. 
Perhaps some kind of chart review on all surgeries?

Thanks

Nita Searcy, HT/HTL (ASCP)
Scott and White Hospital
Temple, Texas
Division Manager, Anatomic Pathology
254-724-2438

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Message: 2
Date: Thu, 2 Dec 2004 11:14:47 -0700
From: "Elizabeth Chlipala" <liz <@t> premierlab.com>
Subject: [Histonet] immunohistochemical marker for osteoblasts
To: "'Histonet'" <histonet <@t> pathology.swmed.edu>
Message-ID: <000001c4d89a$ce98a550$76d48a80 <@t> AMY>
Content-Type: text/plain;	charset="us-ascii"

Is anyone aware of a Immunohistochemical marker that is specific for
osteoblasts?  I have seen references on BAP (bone alkaline phosphatase)
and some other clones such as 2D3 and 2H10, but I have been unable to
find a source.

Thanks

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
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: 3
Date: Thu, 02 Dec 2004 13:35:22 -0500
From: "Fred Underwood" <funderwood <@t> mcohio.org>
Subject: [BULK] - RE: [Histonet] static electricity
To: <histonet <@t> lists.utsouthwestern.edu>,
	<bettina.hutz <@t> orionpharma.com>,	<Laurie.Pereira <@t> sdcounty.ca.gov>
Message-ID: <s1af1a24.097 <@t> mcohio.org>
Content-Type: text/plain; charset=US-ASCII

Static Guard (brand name) aerosol spray works well.  Spray a small
amount on your knife clamp and other "sticky" microtome surfaces.

>>> <bettina.hutz <@t> orionpharma.com> 12/02/04 12:26AM >>>
Hello,

I have the same problem during winter time - and in summer time it is
the
opposite: it is "common accepted" that in July/August there is hardly
any
staining possible, because the huminidy goes immediately to the alcohol
and
then the xylene is milky and mounting not proper. When I have been
working
in Germany for 10 years I never has this effect, but since I work in
Finland
I am fighting with it every summer. So, if anyone has a solution to
the
humidity problem, please share it with me:)

Tina

-----Original Message-----
From: Pereira, Laurie [mailto:Laurie.Pereira <@t> sdcounty.ca.gov] 
Sent: Thursday, December 02, 2004 1:18 AM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] static electricity

Hey all,

I'm new at Histology and am having a problem with static electricity.
During the winter, everything tends to stick to me.  I've heard that a
humidifier works pretty well but our laboratory is pretty large.  Will
a
humidifier work in such a large space?  Does anyone have any hints on
how to
deal with the wonders of static electricity?  Any information would be
extremely helpful!

Thanks

Frustrated

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

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

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

Message: 4
Date: Thu, 02 Dec 2004 18:55:55 +0000
From: hessd101 <@t> comcast.net
Subject: [Histonet] X-gal and pap pens?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<120220041855.7738.41AF653B000591D600001E3A2200761438CECFCE0B9C9C0A08 <@t> comcast.net>

Content-Type: text/plain

I've searched far and wide for a solution to this problem, and I hope the experts can finally help me. Our lab performs x-gal staining on tissue sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we tried to order new pens from Kiyota. Apparently, their clear pens were discontinued because of toxicity, and now the only pen available from them is the Super HT Pap Pen. The new pens put down a flimsy, blue-green colored barrier. The trouble is this - when X-gal staining, some component of the stain reacts with the dye in the pap pen producing a grainy red precipitate which discolors the slide, tissue, etc. In addition, an oily reidue floats up on the solution and can stick to tissue while removing it. In a nutshell, the new pens are awful. Does anyone know of a clear pen that is still sold (EMS, zymed all seem to have colored pens), or have experience with a colored pap pen that doesn't react with xgal staining solution? Thanks

Darren Hess
MD/PhD student, Dept. of Neuroscience
University of Pennsylvania School of Medicine

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

Message: 5
Date: Thu, 2 Dec 2004 14:05:54 -0600
From: "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org>
Subject: [Histonet] RE: fixation for RE: Masson's Trichrome on frozen
	heart	sections
To: "Gayle Callis" <gcallis <@t> montana.edu>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<A61F23E937E4DA488E0C0F60093843D901E497EE <@t> ccetxm030.echristus.net>
Content-Type: text/plain;	charset="iso-8859-1"

After sectioning we put the slide in a coplin jar with alcoholic formalin for 5 minutes, and proceed with the stain.

Juan C. Gutierrez, HT(ASCP)
Histology Laboratory Supervisor
(210)704-2533

My opinions are my own and do not reflect those of my employer.  Long live free speech!

-----Original Message-----
From: Gayle Callis [mailto:gcallis <@t> montana.edu] 
Sent: Thursday, December 02, 2004 11:20 AM
To: GUTIERREZ, JUAN; Histonet <@t> lists.utsouthwestern.edu
Subject: fixation for RE: Masson's Trichrome on frozen heart sections

Juan,

How do you fix fresh tissue frozen sections?  Bouins or NBF?  and fixation 
times?  If we were to do this stain, our tissues are NEVER prefixed before 
snap freezing.

A one step staining procedure makes a lot of sense when handling more 
fragile frozen sections.  I know one substitute light green with aniline 
blue, or vice versa - what ever one likes best.  This was discussed just a 
few months ago on Histonet.

At 09:48 AM 12/2/2004, you wrote:
>We use Gomori's trichrome on frozen sections.  It's a one step procedure 
>that gives you a faster answer.  Are you able to try alternatives to MTS?
>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: 6
Date: Thu, 2 Dec 2004 16:48:43 -0500
From: rgrow <@t> bmnet.com
Subject: [Histonet] RE: static electricity/humidity problems
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<OFB152E812.69AADB18-ON85256F5E.0075F288-85256F5E.0077D129 <@t> bmnet.com>
Content-Type: text/plain; charset=US-ASCII

Tina,

There have been several good responses regarding static.  Try them.  Use
what works best in your lab.

Humidity is a big problem in the southern United States.   Air conditioning
systems and de-humidifiers can't handle the load in old drafty buildings.
We have solved this problem by adding silica gel crystals to our last
absolute alcohol and first xylene in the dehydrating and clearing stations.
This step has been successfully used here for many years.  It  also extends
the life of the reagents 2x.
We order it from:  Sigma Aldrich Company, Catalog #:  21,444.2  Silica Gel
2.5 Kg.
A word of caution: Do dispense and discard carefully as it is a carcinogen.

Good luck.

Renee Grow, BA., HT (ASCP)
rgrow <@t> bmnet.com
Histology Supervisor
Blount Memorial Hospital
907 E. Lamar Alexander Pkwy.
Maryville, TN  37804-5016
(865) 977-4744
(865) 977-5766 Fax

From: bettina.hutz <@t> orionpharma.com [mailto:bettina.hutz <@t> orionpharma.com]
Sent: 02 December 2004 05:26
To: Laurie.Pereira <@t> sdcounty.ca.gov; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] static electricity[Scanned]

Hello,

I have the same problem during winter time - and in summer time it is the
opposite: it is "common accepted" that in July/August there is hardly any
staining possible, because the huminidy goes immediately to the alcohol and
then the xylene is milky and mounting not proper. When I have been working
in Germany for 10 years I never has this effect, but since I work in
Finland
I am fighting with it every summer. So, if anyone has a solution to the
humidity problem, please share it with me:)

Tina

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

Message: 7
Date: Thu, 02 Dec 2004 15:26:32 -0700
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: Re: [Histonet] X-gal and pap pens?
To: hessd101 <@t> comcast.net, Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.1.20041202150331.01b51298 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Darren,

Try the Vector ImmEdge pens, they are very clean, dry nicely, and have long 
since replaced Kiyota's pens in our lab.  Vector sells their pens in 
package of two, at a price of one pen from other vendors.   One thing to do 
when using the Immedge pen - shake the begolly's out of them to disperse 
the active marking goo.  Their technical rep recommended this be done.  I 
actually vortex mine to insure goo redistribution.

  It is interesting that Kiyota's Super Pap Pens were declared toxic.  I 
had wondered about this ( after noticing some very strong solvent odors) 
and asked them about it, but they said they were acceptable, but that fact 
changed somewhere along the line.

DAKO has a clear pen, but be careful when dispensing - if you press toooooo 
hard, you have a Niagara Falls of goo all over everything.  I macerated ( 
with a pliers!) their hard tip to make it a tidge fuzzier to capture the 
goo rush and also pulled a tip from a favorite pen and did a trade of hard 
tip with softer tip.   We also let DAKO pen marks dry longer, but I have 
that luxury of time and with frozen sections before staining and/or 
fixation.  Hopefully DAKO corrected this over-dispensing problem as the goo 
is good and it is clear.

If all else fails, you can buy hydrophobic barrier slides with lines in a 
circle, squares or rectangles that work quite well. Just mount the section 
inside lines and do the staining.  Erie has a huge selection of these, and 
we have even had custom hydrophobic barrier slides made for us to eliminate 
pap pens.  Lovely thing is the barrier stays put unlike the pen marking, 
and the lines we liked were the one that are flatter like the Erie control 
slides so mounting a coverslip is more flush with slide surface. You can 
always talk to Erie about their selection.

At 11:55 AM 12/2/2004, you wrote:
>I've searched far and wide for a solution to this problem, and I hope the 
>experts can finally help me. Our lab performs x-gal staining on tissue 
>sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put 
>down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we 
>tried to order new pens from Kiyota. Apparently, their clear pens were 
>discontinued because of toxicity, and now the only pen available from them 
>is the Super HT Pap Pen. The new pens put down a flimsy, blue-green 
>colored barrier. The trouble is this - when X-gal staining, some component 
>of the stain reacts with the dye in the pap pen producing a grainy red 
>precipitate which discolors the slide, tissue, etc. In addition, an oily 
>reidue floats up on the solution and can stick to tissue while removing 
>it. In a nutshell, the new pens are awful. Does anyone know of a clear pen 
>that is still sold (EMS, zymed all seem to have colored pens), or have 
>experience with a colored pap pen that doesn'!
>  t react with xgal staining solution? Thanks
>
>Darren Hess
>MD/PhD student, Dept. of Neuroscience
>University of Pennsylvania School of Medicine
>_______________________________________________
>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)

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

Message: 8
Date: Fri, 3 Dec 2004 09:46:08 +1100
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] Destaining DifQuik
To: "'Mary North'" <northma <@t> ohsu.edu>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E12B <@t> simba.kids>
Content-Type: text/plain

To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol.

If the smears have been air-dried prior to DifQuik staining, then PAP
staining will be very dissapointing since the cells will be affected by
air-drying. There is no technique known to reverse this.

Regards

Tony Henwood JP, 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: Mary North [mailto:northma <@t> ohsu.edu] 
Sent: Friday, 3 December 2004 3:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Destaining DifQuik

Has anyone tried to destain DifQuik-stained cytology preps so that it
can be restained with PAP?
Mary North, HT(ASCP), HTL
OHSU
Portland, OR

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

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

Message: 9
Date: Thu, 2 Dec 2004 23:05:01 EST
From: WWmn916 <@t> aol.com
Subject: [Histonet] C4D Vendor
To: histonet <@t> pathology.swmed.edu
Message-ID: <88.1affec28.2ee13fed <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"

Greetings everyone,

Has anyone heard if there is a C4D vendor?  Is there  anything on the horizon 
for availability?

Next question.....it may be simple, but does HIPAA rules say patient names  
can not be on slides?

Thanks,
Deb King, HT(ASCP)    
Sacramento, CA

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

Message: 10
Date: Fri, 3 Dec 2004 09:51:35 +0200 
From: bettina.hutz <@t> orionpharma.com
Subject: RE: [Histonet] RE: static electricity/humidity problems
To: rgrow <@t> bmnet.com, histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<243E1A79AF5D89438C70513DBADAA4F0019E3FFD <@t> sfies-exchange1.orionnet.org>

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

Hello Renee,

this sounds good, I WILL try it:)
Thanks a lot, "kiitos" as we say in finnish:)
Tina

-----Original Message-----
From: rgrow <@t> bmnet.com [mailto:rgrow <@t> bmnet.com] 
Sent: Thursday, December 02, 2004 11:49 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: static electricity/humidity problems

Tina,

There have been several good responses regarding static.  Try them.  Use
what works best in your lab.

Humidity is a big problem in the southern United States.   Air conditioning
systems and de-humidifiers can't handle the load in old drafty buildings. We
have solved this problem by adding silica gel crystals to our last absolute
alcohol and first xylene in the dehydrating and clearing stations. This step
has been successfully used here for many years.  It  also extends the life
of the reagents 2x. We order it from:  Sigma Aldrich Company, Catalog #:
21,444.2  Silica Gel 2.5 Kg. A word of caution: Do dispense and discard
carefully as it is a carcinogen.

Good luck.

Renee Grow, BA., HT (ASCP)
rgrow <@t> bmnet.com
Histology Supervisor
Blount Memorial Hospital
907 E. Lamar Alexander Pkwy.
Maryville, TN  37804-5016
(865) 977-4744
(865) 977-5766 Fax

From: bettina.hutz <@t> orionpharma.com [mailto:bettina.hutz <@t> orionpharma.com]
Sent: 02 December 2004 05:26
To: Laurie.Pereira <@t> sdcounty.ca.gov; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] static electricity[Scanned]

Hello,

I have the same problem during winter time - and in summer time it is the
opposite: it is "common accepted" that in July/August there is hardly any
staining possible, because the huminidy goes immediately to the alcohol and
then the xylene is milky and mounting not proper. When I have been working
in Germany for 10 years I never has this effect, but since I work in Finland
I am fighting with it every summer. So, if anyone has a solution to the
humidity problem, please share it with me:)

Tina

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

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

Message: 11
Date: Fri, 3 Dec 2004 08:13:48 -0000 
From: Kemlo Rogerson <Kemlo.Rogerson <@t> elht.nhs.uk>
Subject: RE: [Histonet] Destaining DifQuik[Scanned]
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B56F <@t> bhrv-nt-11.bhrv.nwest.nhs.uk>

Content-Type: text/plain

Diff-Quick is usually carried out on air-dried preps and PAP on alcohol
fixed. You can destain Diff-quick in 1% acid alcohol or just by leaving in
alcohol, but the PAP will be hopeless as it was air-dried.

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194

-----Original Message-----
From: Mary North [mailto:northma <@t> ohsu.edu] 
Sent: 02 December 2004 16:40
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Destaining DifQuik[Scanned]

Has anyone tried to destain DifQuik-stained cytology preps so that it
can be restained with PAP?
Mary North, HT(ASCP), HTL
OHSU
Portland, OR

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

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

Message: 12
Date: Fri, 3 Dec 2004 08:22:12 -0000 
From: Kemlo Rogerson <Kemlo.Rogerson <@t> elht.nhs.uk>
Subject: RE: [Histonet] Destaining DifQuik[Scanned]
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B571 <@t> bhrv-nt-11.bhrv.nwest.nhs.uk>

Content-Type: text/plain

Oops sorry, you already said it! That's the trouble being in the UK, life
has passed you by as you sleep.

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194

-----Original Message-----
From: Tony Henwood [mailto:AnthonyH <@t> chw.edu.au] 
Sent: 02 December 2004 22:46
To: 'Mary North'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Destaining DifQuik[Scanned]

To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol.

If the smears have been air-dried prior to DifQuik staining, then PAP
staining will be very dissapointing since the cells will be affected by
air-drying. There is no technique known to reverse this.

Regards

Tony Henwood JP, 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: Mary North [mailto:northma <@t> ohsu.edu] 
Sent: Friday, 3 December 2004 3:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Destaining DifQuik

Has anyone tried to destain DifQuik-stained cytology preps so that it
can be restained with PAP?
Mary North, HT(ASCP), HTL
OHSU
Portland, OR

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

**********************************************************************
This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom they
are addressed. If you are not the intended recipient, please 
delete it and notify the sender.

Views expressed in this message and any attachments are those 
of the individual sender, and are not necessarily the views of The
Children's Hospital at Westmead

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

Message: 13
Date: Fri, 3 Dec 2004 08:31:14 -0000 
From: Kemlo Rogerson <Kemlo.Rogerson <@t> elht.nhs.uk>
Subject: RE: [Histonet] M.tuberculosis in formalin-fixed
	tissue[Scanned]
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B572 <@t> bhrv-nt-11.bhrv.nwest.nhs.uk>

Content-Type: text/plain

No-one replied to this yet? No?

Ok. TB lives (hibernates) in bone, that's why, in the old days people were
sent to Sanatoriums. They recovered from their TB, by good diet etc., but
when they were released and their diet/ health got worse, the bone
de-ossified and the TB was released; they then suffered a relapse.

I assume that can happen in lung etc., as the TB may be encapsulated in
areas of bony spiculues (can't spell that) it is shielded from the fixative.
I assume staining, cutting, etc., can release the bacteria from its bony
coffin to make the sections infective.

I would assume the risk is low, but there all the same. There is an
'explosion' of TB in London, I gather from the News today.

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194

-----Original Message-----
From: Therersa Stegall [mailto:STEGTM <@t> samcstl.org] 
Sent: 02 December 2004 17:07
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] M.tuberculosis in formalin-fixed tissue[Scanned]

Say y'all.  I have a copy of CAP Today in front of me, last months'. 
There's an article on pg118 that says somewhere around 10% of cultured
lung tissue, fixed in formalin, grew M.tuberculosis.  We all assume the
risk when cutting frozens, and the subsequent decontamination
procedures; has anyone really thought about this?  Have you heard
anything else about such?  Does anyone out there assume that this fixed
tissue is still viable (we assume that it is a mere capsular ghost of
itself).  I'd be interested to hear how you deal with this type of
tissue; precautions or not...... Terre

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

Message: 14
Date: Fri, 3 Dec 2004 14:34:53 +0530
From: "Arvind Pundir" <arvind <@t> nbrc.res.in>
Subject: [Histonet] cryosection fetal tissue
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A704 <@t> mail.nbrc.res.in>
Content-Type: text/plain; charset="utf-8"

can any one pls tell how to embed and cut  human fetal tissue for immuno as it is too soft to cut 

thanks in advance

Arvind Singh Pundir

National Brain Research Centre
Near NSG Campus
Manesar, Haryana - 122 050, India

arvind <@t> nbrc.res.in

Tel.: 91-124 âEUR" 233 8922-26
Fax: 91-124 - 233 89 10 / 91-124 - 233 89 28 aNational Brain Research Centre 
Manesar, Gurgaon DiNational Brain Research Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 
st. Haryana-122 050, India 
rch Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 
National Brain Research Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 
National Brain Research Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 
National Brain Research Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 
National Brain Research Centre 
Manesar, Gurgaon Dist. Haryana-122 050, India 

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

Message: 15
Date: Fri, 3 Dec 2004 05:37:58 -0500
From: <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
	solutions[Scanned]
To: "Kemlo Rogerson" <Kemlo.Rogerson <@t> elht.nhs.uk>,	"'Jackie M
	O'Connor'" <Jackie.O'Connor <@t> abbott.com>,	"Johnson, Teri"
	<TJJ <@t> Stowers-Institute.org>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <004d01c4d924$29272da0$2329d445 <@t> domainnotset.invalid>
Content-Type: text/plain;	charset="iso-8859-1"

Let's see if I can explain pH, hematoxylin, red vs. blue, and bluing, and
not get myself into a murky mess of chemistry.

Hematoxylin solution is actually an aluminum salt and oxidized hematoxylin,
collectively known as aluminum-hematein (Al-hematein).

In order to stain just the DNA and RNA, the pH of the Al-hematein solution
needs to be between 2.2 to 2.9 (most usually around 2.4-2.5).

At this lower (acidic) pH, there are a lot of hydrogen ions (H+) in the
Al-Hematein solution. These excess H+ bind to the Al-hematein, causing a
shift to the red.

When the stained tissue is placed in a solution more alkaline than the
2.2-2.9 stain, there are fewer H+ in the staining solution. This removes the
H+ from the Al-Hematein. This causes a color shift in the dye to the blue
range (i.e.,  "bluing the slides").

Therefore, bluing agents must have a pH higher than the mid-2.5. The higher
the pH (more alkaline), the faster the H+ are removed from the Al-Hematein,
and the faster the "bluing" will take place. Bluing at about pH 5.0 can take
up to about 10 minutes to "blue" totally (all red nuclei to some red/some
blue (= purple looking tissue) to all blue nuclei). At pH 10, it will take
30 seconds.

Tap water is more alkaline than the Al-hematein, so it can be used to blue
slides. If the tap water's pH is around 7, this should take about 5 minutes.
Some lab's water is more alkaline than other labs, so will take less time.
The pH of tap water can change depending upon what chemicals the water
treatment facility is using that day to purify the water, so using just tap
water to blue may have some variability of time.

Most labs rely on an alkaline solution (dilute ammonia, dilute lithium
carbonate, Scott's tap water) to speed up the bluing processing and to not
rely upon the variability of water pH.

Hope this helps.

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

----- Original Message -----
From: "Kemlo Rogerson" <Kemlo.Rogerson <@t> elht.nhs.uk>
To: "'Jackie M O'Connor'" <Jackie.O'Connor <@t> abbott.com>; "Johnson, Teri"
<TJJ <@t> Stowers-Institute.org>
Cc: "Histonet" <histonet <@t> lists.utsouthwestern.edu>;
<histonet-bounces <@t> lists.utsouthwestern.edu>
Sent: Wednesday, December 01, 2004 3:18 AM
Subject: RE: [Histonet] Re: Ammonia water pH and other bluing
solutions[Scanned]

> I think you are wrong, so I'll correct you, as you requested.
>
> I thought the 'blue' state of haematin was, like litmus, the effect an
> alkaline condition has on the siderphilic dye. Red, like litmus, denoted
> acidic conditions. Damned if I know what colour (with the 'u') haematin
goes
> when neutral (bluey/ red?)
>
> Running tap water takes longer, I concede, but good things are worth
waiting
> for and it's easier to make up; we rush too much I find. TIP: Never use
> London water, been through too many kidneys!
>
> -----Original Message-----
> From: Jackie M O'Connor [mailto:Jackie.O'Connor <@t> abbott.com]
> Sent: 30 November 2004 19:09
> To: Johnson, Teri
> Cc: Histonet; histonet-bounces <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
> solutions[Scanned]
>
> Anyone remember using saturated lithium carbonate?  I could probably use a
> little lithium right now . . . . .
>
> Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just
> bringing the slides BACK to a neutral pH after treating them with acid
> which makes them purple-ish?  I like the ammonia because it 'seems' to
> make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to
> make them "blah".(Perception is in the eye of the beholder)   Running tap
> water will make the same miracle happen, but it takes longer.
>
>
>
>
>
>
> "Johnson, Teri" <TJJ <@t> Stowers-Institute.org>
> Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
> 11/30/2004 12:51 PM
>
>
>         To:     "Histonet" <histonet <@t> lists.utsouthwestern.edu>
>         cc:
>         Subject:        [Histonet] Re: Ammonia water pH and other bluing
> solutions
>
>
> Way back in the day, Brigati was using PBS on his immunostainer to blue
> after the hematoxylin counterstain.  I have since used PBS to blue my
> slides instead of ammonium hydroxide/water.  It's cheap, and very easy
> on the sections, and I have plenty of it on hand.
>
>
> Teri Johnson
> Managing Director Histology Facility
> Stowers Institute for Medical Research
> 1000 E. 50th St.
> Kansas City, Missouri  64110
> tjj <@t> stowers-institute.org
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
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>
>
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------------------------------

Message: 16
Date: Fri, 3 Dec 2004 10:47:26 -0000 
From: Kemlo Rogerson <Kemlo.Rogerson <@t> elht.nhs.uk>
Subject: RE: [Histonet] Re: Ammonia water pH and other bluing
	solutions[Sc	anned]
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B573 <@t> bhrv-nt-11.bhrv.nwest.nhs.uk>

Content-Type: text/plain

Oh I see now, so why are nuclei blue?

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194

-----Original Message-----
From: lpwenk <@t> sbcglobal.net [mailto:lpwenk <@t> sbcglobal.net] 
Sent: 03 December 2004 10:38
To: Kemlo Rogerson; 'Jackie M O'Connor'; Johnson, Teri
Cc: Histonet
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
solutions[Scanned]