[Histonet] Re: Histonet Digest, Vol 58, Issue 12 Recyclers

Wed Sep 10 13:56:56 CDT 2008

I used a CBG recycler for years.....many years and absolutely loved it!  Worked great, excellent recovery (xylene) and minimal maintenance.
Great (people) support from the company too.

Jeanne

Jeanne Clark HT/MLT (ASCP)
Pathology Manager
Mission Hospitals
Asheville, NC
423-612-1213

--- On Wed, 9/10/08, histonet-request <@t> lists.utsouthwestern.edu <histonet-request <@t> lists.utsouthwestern.edu> wrote:

From: histonet-request <@t> lists.utsouthwestern.edu <histonet-request <@t> lists.utsouthwestern.edu>
Subject: Histonet Digest, Vol 58, Issue 12
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, September 10, 2008, 1:21 PM

Send Histonet mailing list submissions to
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Today's Topics:

   1. Manual Paraffin Embedding (Merced Leiker)
   2. Manual Paraffin Embedding (Paula Pierce)
   3. Re: Manual Paraffin Embedding (Bryan Llewellyn)
   4. Re: Histonet Digest, Vol 58, Issue 8 (Cheryl Crowder)
   5. Re: methylene blue (Rene J Buesa)
   6. Re: Manual Paraffin Embedding (Rene J Buesa)
   7. Re: Manual Paraffin Embedding (Emily Sours)
   8. Paraformaldehyde (Mary P. Brownson)
   9. RE: Manual Paraffin Embedding (Kemlo Rogerson)
  10. Histology Positions (Cheri Miller)
  11. Help (RENEE FISHER)
  12. please un subscribe (RENEE FISHER)
  13. Recyclers (Bauer, Karen)

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

Message: 1
Date: Tue, 09 Sep 2008 16:54:43 -0400
From: Merced Leiker <leiker <@t> buffalo.edu>
Subject: [Histonet] Manual Paraffin Embedding
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <6089D76726392CD3B3399BFC <@t> bchwxp2702.ad.med.buffalo.edu>
Content-Type: text/plain; charset=us-ascii; format=flowed

Does anyone process and embed tissues manually instead of using automated 
and expensive equipment?  Can you tell me how you do it?  Thanks.

Merced

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

Message: 2
Date: Tue, 9 Sep 2008 14:19:57 -0700 (PDT)
From: Paula Pierce <contact <@t> excaliburpathology.com>
Subject: [Histonet] Manual Paraffin Embedding
To: Merced Leiker <leiker <@t> buffalo.edu>,	Histonet
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <548890.39203.qm <@t> web1116.biz.mail.sk1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

I sometimes still use a paraffin dispenser for really large blocks. It looks
like a big coffee maker, like caterers use. You will still need molds. 
Embedding centers work on the same principle, they just incorporate the melted
paraffin bay and dispenser with a cold plate on the side.
Paula Pierce

----- Original Message ----
From: Merced Leiker <leiker <@t> buffalo.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, September 9, 2008 3:54:43 PM
Subject: [Histonet] Manual Paraffin Embedding

Does anyone process and embed tissues manually instead of using automated 
and expensive equipment?  Can you tell me how you do it?  Thanks.

Merced

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Message: 3
Date: Tue, 9 Sep 2008 15:21:02 -0700
From: "Bryan Llewellyn" <llewllew <@t> shaw.ca>
Subject: Re: [Histonet] Manual Paraffin Embedding
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <9A9BD3D0B0284D23904E676EA082BD4F <@t> Compaq>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
	reply-type=response

I don't do this anymore, nor for 40 years now, but this is what we used to 
do aeons ago.

1. Fix in 10% NBF for 48 hours.
2 Rinse off excess with tap water for 1 minute.
3. Select pieces of tissue with maximum dimensions of 2cm x 1.5 cm x 0.3 cm.
4. Place into cassettes if you have them, then into a large container.  If 
you do not have cassettes, place into small jars, each case in a different 
jar.  Place a label in each cassette or each jar with the case ID.
5. Cover the tissue or cassettes with 70% ethanol, methylated spirits or 
isopropanol, agitate gently.  Leave overnight.
6. Next morning, replace the 70& alcohol with 85% alcohol, leave for the 
day, agitating gently periodically.
7. Before leaving in the evening, replace the alcohol with 95% alcohol, 
agitate gently and leave overnight.
8. Next morning, replace the alcohol with 100% alcohol, gently agitate 
periodically.  Repeat at noon and before you 9. leave for the evening, 
gently agitating.
10. Next morning, replace the alcohol with clearant, preferably xylene or 
toluene.  Leave for one hour, gently agitating periodically.
11. Repeat the clearant twice more, agitate gently.
12. Place into premelted paraffin wax for 1 hour at 65C.  Check periodically 
and when all congealed wax has remelted, agitate gently.
13. Repeat at least twice more, preferably under vacuum - not too strong. 
Some technologists used to leave the final change overnight.  Doing so 
doesn't do much harm and improves penetration.  Agitation can't be done

under vacuum, so release the vacuum periodically, agitate and reapply it.
14. Block out into molds.  (If you do not have molds, use a metal - tinned 
steel or aluminum - lid with a depth of 1cm.  LIGHTLY coat it with glycerol 
first.)  Place a thin (3mm) layer of hot wax in the mold and place the 
tissue, with the surface to be sectioned down, into it.  Top up the mold 
with wax so there is at least 2-3 mm wax over the top of the tissue.  Put 
the ID label conspiciously next to the tissue.  Do NOT block out more than 
one tissue or case without placing the ID labels, this WILL lead to serious 
identification errors.  Do all this by the oven and keep the door closed as 
much as possible.  Work fast so that the wax around the tissues does not 
begin to congeal as that causes problems during sectioning and floating out. 
You must get the wax around the tissue and the wax in the mold to blend 
completely, so if the wax congeals around the tissue, leave it to remelt 
before blocking out.  Many of us used to keep a bunsen burner alight and 
flame the top of the molds to keep the wax molten - a practice probably 
considered unsafe now.
15. Allow the wax in the mold to skin over thoroughly, then GENTLY lower 
into cold tap water to cool.
16. When completely cold and solid, use a heavy knife to score and trim the 
wax blocks.
17. To section, melt the trimmed block onto the block holder.

Bryan Llewellyn

----- Original Message ----- 
From: "Merced Leiker" <leiker <@t> buffalo.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, September 09, 2008 1:54 PM
Subject: [Histonet] Manual Paraffin Embedding

> Does anyone process and embed tissues manually instead of using automated 
> and expensive equipment?  Can you tell me how you do it?  Thanks.
>
> Merced
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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

Message: 4
Date: Tue, 09 Sep 2008 17:54:41 -0500
From: "Cheryl Crowder" <ccrowder <@t> vetmed.lsu.edu>
Subject: [Histonet] Re: Histonet Digest, Vol 58, Issue 8
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <WorldClient-F200809091754.AA54413878 <@t> vetmed.lsu.edu>
Content-Type: text/plain; charset="us-ascii"

Maxim - You sent me an e-mail in August.  I replied several times, but the 
e-mail did not go through.  Is your address complete or is there another I 
can  use?

Cheryl Crowder, BA, HTL(ASCP)
Chief Technologist
Anatomic Pathology
Department of Pathobiological Sciences
School of Veterinary Medicine
Louisiana State University
Skip Bertman Drive
Baton Rouge, LA  70803

225-578-9734
FAX:  225-578-9720

-----Original Message-----

From: histonet-request <@t> lists.utsouthwestern.edu

To: histonet <@t> lists.utsouthwestern.edu

Date: Sun, 07 Sep 2008 12:02:56 -0500

Subject: Histonet Digest, Vol 58, Issue 8

Send Histonet mailing list submissions to

   histonet <@t> lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit

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

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   histonet-request <@t> lists.utsouthwestern.edu

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than "Re: Contents of Histonet digest..."

Today's Topics:

   1. RE: reagent question (Maxim_71 <@t> mail.ru)

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

Message: 1

Date: Sat, 6 Sep 2008 23:38:42 +0400

From: Maxim_71 <@t> mail.ru

Subject: RE: [Histonet] reagent question

To: pathology.histology <@t> gmail.com

Cc: histonet <@t> lists.utsouthwestern.edu

Message-ID: <03082354.20080906233842 <@t> mail.ru>

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

More than one year we uses isopropanol as dehydratant and enjoy for

quality of specimens, which processed with this reagent.

It is very useful for bone, cartillage, uterus, breast, colon,

skin, eyeball and other difficult specimens.

Some unpleasant odor of "cat urine" does not disturb to use it in

manual processing.

We believes, that will be time, when we will have a vacuum

processor...

For slides we use acetone as H&E as SS. We stain all our slides

manually.

Maxim Peshkov,

Russia,

Taganrog.                          mailto:Maxim_71 <@t> mail.ru

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End of Histonet Digest, Vol 58, Issue 8

***************************************

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

Message: 5
Date: Wed, 10 Sep 2008 05:00:17 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] methylene blue
To: histonet <@t> lists.utsouthwestern.edu,	"Perry, Margaret"
	<Margaret.Perry <@t> sdstate.edu>
Message-ID: <285564.59557.qm <@t> web65708.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Methylene blue does not require anything special, this stain never fails. Just
hydrate your section, stain with 1% aq. methBlue for 5 minutes, wash and
dehydrate QUICKLYLY cover and that is all.<René J.

--- On Tue, 9/9/08, Perry, Margaret <Margaret.Perry <@t> sdstate.edu> wrote:

From: Perry, Margaret <Margaret.Perry <@t> sdstate.edu>
Subject: [Histonet] methylene blue
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, September 9, 2008, 4:22 PM

Help!     We have been asked to do a methylene blue stain for
Myxosporidia and don't have a procedure.  Of course the pathologist
wants it quickly.  I can't get into the archives.  It must be down right
now.  Can someone help us?

Margaret Perry HT (ASCP)

IHC Lab Manager Veterinary Science

Animal Disease Research and Diagnostic Lab

South Dakota State University

Box 2175 North Campus Drive

Brookings SD 57007

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

Message: 6
Date: Wed, 10 Sep 2008 05:09:14 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Manual Paraffin Embedding
To: histonet <@t> lists.utsouthwestern.edu, Merced Leiker
	<leiker <@t> buffalo.edu>
Message-ID: <577961.55420.qm <@t> web65715.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Merced:
Processing and embedding tissues manually is still done occasionally here in
the US and specially abroad. As a matter of fact 2% us US labs and 14% of
foreign labs routinely manually process their tissues.

It implies running the tissues through all the dehydration, clearing and
infiltration steps to finally prepare the blocks, also manually, using melted
paraffin and dispensed in paper molds, or using Leuckhart rectangles. 
The description would be very long for an e-mail, so my advise is to get a
histotechnique book, like Bolles-Lee's "Microtomist Vade-Mecum",
or Peter Gray's "The Microtomist's formulary and guide", both
are for sale at Amazon.com/books or you may find a copy at the University of
Buffalo.
René J.

--- On Tue, 9/9/08, Merced Leiker <leiker <@t> buffalo.edu> wrote:

From: Merced Leiker <leiker <@t> buffalo.edu>
Subject: [Histonet] Manual Paraffin Embedding
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, September 9, 2008, 4:54 PM

Does anyone process and embed tissues manually instead of using automated 
and expensive equipment?  Can you tell me how you do it?  Thanks.

Merced

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

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

Message: 7
Date: Wed, 10 Sep 2008 09:31:17 -0400
From: "Emily Sours" <talulahgosh <@t> gmail.com>
Subject: Re: [Histonet] Manual Paraffin Embedding
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<b39794b0809100631q673b0067ue072d94e325b7495 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

We just use melted paraffin in a vacuum oven.  The paraffin is kept at 60C
in small glass petri dishes and is moved from dish to dish every half hour
for about 3 hours.  Then you fill a plastic mold halfway, let the paraffin
cool until it's hard enough to pour more on without it melting again, but
not fully cooled, put in your tissue, cover it with melted paraffin and let
it cool overnight.
You don't need the vacuum oven, it just gets rid of bubbles in the
paraffin.
You also need to use heated instruments (we just warm them briefly in an
alcohol lamp flame) when transferring the embryo in paraffin.

Emily
-- 
An overcivilized people grow complacent and careless and leave the door open
for a tribe of fanatical savages, through a mixture of luck, treachery, and
the foulest inhumanity, to usurp their place for a few years.
-Richard Adams, "Shardik", 1974

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

Message: 8
Date: Tue, 9 Sep 2008 14:50:33 -0600
From: "Mary P. Brownson" <mpb1211 <@t> uwyo.edu>
Subject: [Histonet] Paraformaldehyde
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<85F291B637727346BA8274B0A43C24FF02946E2C <@t> TELEGRAPH1.uwyo.edu>
Content-Type: text/plain;	charset="us-ascii"

Hello,

I plan to use paraformaldehyde for perfusion of rat brains.  My question
is; what grade paraformaldehde should I be using?  Reagent grade, 95% or
other?  Or does it not matter?

Thank you,

Mary Brownson

School of Pharmacy

University of Wyoming

Laramie, WY 82070

mpb1211 <@t> uwyo.edu

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

Message: 9
Date: Wed, 10 Sep 2008 15:10:35 +0100
From: "Kemlo Rogerson" <Kemlo.Rogerson <@t> waht.swest.nhs.uk>
Subject: RE: [Histonet] Manual Paraffin Embedding
To: "Merced Leiker" <leiker <@t> buffalo.edu>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<86ADE4EB583CE64799A9924684A0FBBF054834F2 <@t> wahtntex2.waht.swest.nhs.uk>
Content-Type: text/plain;	charset="us-ascii"

Does anyone process and embed tissues manually instead of using
automated 
and expensive equipment?  Can you tell me how you do it?  Thanks.

Merced

When I was a lad I used to manual process Brains and other stuff. In
retrospect I wonder why we use the same processing schedules for
disparate organs. We ought to have a kidney schedule and a heart
schedule specifically for each organ type. I always knew a tissue was
processed adequately as it changed in its translucency. If the was water
still present in the tissue you could see its presence in the clearing
agent and smell clearing agent in the wax. However you exposed yourself
to carcinogenic vapours and I guess that it is right and proper we
automated and contained. I wonder if that's why I've grown two heads? 

Kemlo Rogerson        
Pathology Manager
DD   01934 647057 or extension 3311
Mob 07749 754194; Pager 07659 597107;
Don't be afraid to take a big step when one is indicated. You can't
cross a chasm in two small jumps. --Buckminster Fuller 

This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
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its contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
Thank you for your co-operation

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

Message: 10
Date: Wed, 10 Sep 2008 10:06:56 -0500
From: "Cheri Miller" <cmiller <@t> physlab.com>
Subject: [Histonet] Histology Positions
To: <histonet <@t> lists.utsouthwestern.edu>
Cc: <histonet-bounces <@t> lists.utsouthwestern.edu>
Message-ID: <000001c91356$de4ccc60$3d02a8c0 <@t> plab.local>
Content-Type: text/plain;	charset="us-ascii"

Is there anyone looking for a Histotech or a Histology Supervisor in the
Omaha, Council Bluffs or Lincoln area??  Let me know. Thanks, Cheri

Cheryl Miller HT (ASCP)

Histology Supervisor

Physicians Laboratory,P.C.

Omaha, Ne. 

402 738 5052

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Message: 11
Date: Wed, 10 Sep 2008 12:16:54 -0400
From: "RENEE FISHER" <RFISHER <@t> gbmc.org>
Subject: [Histonet] Help
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <48C7BAB5.5C87.00CA.0 <@t> gbmc.org>
Content-Type: text/plain; charset=US-ASCII

please un scribe 
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Message: 12
Date: Wed, 10 Sep 2008 12:19:12 -0400
From: "RENEE FISHER" <RFISHER <@t> gbmc.org>
Subject: [Histonet] please un subscribe
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <48C7BB3F.5C87.00CA.0 <@t> gbmc.org>
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please un subscribe

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Message: 13
Date: Wed, 10 Sep 2008 11:58:09 -0500
From: "Bauer, Karen" <Bauer.Karen <@t> mayo.edu>
Subject: [Histonet] Recyclers
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<FAEEA161EC98C6439D4892E161BCE18A03A1C1B1 <@t> LMMAILVS1.ad.lmmhs.org>
Content-Type: text/plain;	charset="iso-8859-1"

Hello,

I'm in the market to demo some recyclers.  Any vendors are welcome to
contact me personally.  I've already contacted CBG and BR Instruments, so
vendors from those areas need not reply to this message.

Anyone on Histonet that would like to give me their likes and dislikes of
certain recyclers, I'm all ears.

Thanks much,

Karen

Karen L. Bauer HT(ASCP)
Department of Pathology
Histology Section Chief
Luther Hospital
Eau Claire, WI
715-838-3205
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End of Histonet Digest, Vol 58, Issue 12
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