[Histonet] processor options?
Featherstone, Annette
af46 <@t> buffalo.edu
Thu Oct 6 12:15:07 CDT 2011
Maybe someone can come up with a tissue processor that will have an option that will drain the paraffin after a time limit and let tissue cassettes harden in a cooled retort. This would be invaluable to all conscientious histotechs who have tried to make it in to embed when the weather has shut down roads for extended periods of time. Or if you are a "lone" technician who has a family emergency and cannot get in to take the tissue off the machine. How about it, Leica, Sakura, Thermo?
-----Original Message-----
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Sent: Thursday, October 06, 2011 12:58 PM
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Subject: Histonet Digest, Vol 95, Issue 9
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Today's Topics:
1. Re: Tissue left in processor (Rene J Buesa)
2. RE: Tissue left in processor (Sherwood, Margaret)
3. SV: [Histonet] Difference between Neutral Buffered Formalin
10% and NBformaldehyde 3.7 to 4% (Erikstad, Gudrun Hovstein)
4. Re: (no subject) (Angela Bitting)
5. RE: Difference between Neutral Buffered Formalin 10%
andNBformaldehyde 3.7 to 4% (Breeden, Sara)
6. Ken Marissael is out of the office (Ken_Marissael <@t> vwr.com)
7. RE: Floaters (Morken, Timothy)
8. RE: (no subject) (Janice Mahoney)
9. "formol" (Gudrun Lang)
10. control for Hemoglobin? (Kelly Cross)
11. RE: Tissue left in processor (Janice Mahoney)
12. Imidazole buffer (Tammy Pickles)
13. RE: DAPI on thicker sections, problem with gradient
(gayle callis)
14. RE: RE: DAPI on thicker sections, problem with gradient
(Helen Fedor)
15. RE: Cutting frozen sections from visceral fat (idimitro <@t> mun.ca)
16. CMV/HSV Protocols (Dessoye, Michael J)
17. RE: Floaters (joelle weaver)
18. RE: Difference between Neutral Buffered Formalin 10%
andNBformaldehyde 3.7 to 4% (joelle weaver)
19. RE: Tissue left in processor (gayle callis)
----------------------------------------------------------------------
Message: 1
Date: Thu, 6 Oct 2011 07:13:54 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Tissue left in processor
To: "Histonet \(histonet <@t> lists.utsouthwestern.edu\)"
<histonet <@t> lists.utsouthwestern.edu>, RogerCharles <rcharles <@t> pa.gov>
Message-ID:
<1317910434.70026.YahooMailClassic <@t> web65710.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
I do not think that a well fixed, well processed tissue left in molten paraffin for 2 hours after the processor finished will have any adverse outcome.
Ren? J.
--- On Thu, 10/6/11, Charles, Roger <rcharles <@t> pa.gov> wrote:
From: Charles, Roger <rcharles <@t> pa.gov>
Subject: [Histonet] Tissue left in processor
To: "Histonet (histonet <@t> lists.utsouthwestern.edu)" <histonet <@t> lists.utsouthwestern.edu>
Date: Thursday, October 6, 2011, 10:05 AM
Hi All,
Is there any standard on how long tissue cassettes can remain in the processor after processing before the tissue is subjected to unwanted outcomes?? And if so? what type of artifacts can one expect from tissue that was in the processor in molten paraffin for 2 hours after the processing was completed?
Thanks so much.
Roger
Roger Charles| Microbiologist II
Pennsylvania Veterinary Laboratory
2305 North Cameron Street | Harrisburg, PA 17110
Phone: 717.787.8808 | Fax: 717.772.3895
www.agriculture.state.pa.us<http://www.agriculture.state.pa.us>
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------------------------------
Message: 2
Date: Thu, 6 Oct 2011 10:17:43 -0400
From: "Sherwood, Margaret" <MSHERWOOD <@t> PARTNERS.ORG>
Subject: RE: [Histonet] Tissue left in processor
To: "Charles, Roger" <rcharles <@t> pa.gov>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9708DB5A8C <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
I don't think you want to leave it in the processor for 2 hours. Even at the
embedding center, if someone can't come to help orient the tissue, we then take
the cassettes out of the paraffin station and leave on a paper towel. Have done
that for sometimes 2-3 days.
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Charles, Roger
Sent: Thursday, October 06, 2011 10:05 AM
To: Histonet (histonet <@t> lists.utsouthwestern.edu)
Subject: [Histonet] Tissue left in processor
Hi All,
Is there any standard on how long tissue cassettes can remain in the processor
after processing before the tissue is subjected to unwanted outcomes? And if so
what type of artifacts can one expect from tissue that was in the processor in
molten paraffin for 2 hours after the processing was completed?
Thanks so much.
Roger
Roger Charles| Microbiologist II
Pennsylvania Veterinary Laboratory
2305 North Cameron Street | Harrisburg, PA 17110
Phone: 717.787.8808 | Fax: 717.772.3895
www.agriculture.state.pa.us<http://www.agriculture.state.pa.us>
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Message: 3
Date: Thu, 6 Oct 2011 16:22:55 +0200
From: "Erikstad, Gudrun Hovstein" <Gudrun.Erikstad <@t> stolav.no>
Subject: SV: [Histonet] Difference between Neutral Buffered Formalin
10% and NBformaldehyde 3.7 to 4%
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<D3D3E3C2F409474DA3E8CAD7C5B635A6016999E6 <@t> TREXCHVS01.stolav.helsemn.no>
Content-Type: text/plain; charset="iso-8859-1"
I suppose Ren? meant to say:
Formalin is a 37-40% solution of formaldehyde.
10% formalin is a 3,7-4,0% solution of formaldehyde.
I've never heard the use of "Formol", but then again, I'm scandinavian, not som much european... ;-)
Gudrun, Norway
-----Opprinnelig melding-----
Fra: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] P? vegne av Rene J Buesa
Sendt: 6. oktober 2011 15:13
Til: histonet <@t> lists.utsouthwestern.edu; Jenny Vega
Emne: [Histonet] Difference between Neutral Buffered Formalin 10% and NBformaldehyde 3.7 to 4%
Jenny:
Formaldehyde is?the GAS methanal?that dissolves in water at a maximum concentration between 37-40% vol./vol. To prevent its polymerization, small amounts of mathanol (an alcohol)are added.
The 10% solution formaldehyde contains between 3.7 and 4.0% of formaldehyde and is called "formalin" in many countries and "formol" in Europe.
So formalin is a 3.7 to 4.0% solution of formaldehyde.
So a 10% formalin = 3.7-4.0% formaldehyde.
NBF is prepared by added phosphate salts to prevent the formalin to acidify and is used to fix tissues.
It is a pitty that your supervisor has so little knowledge about the area she is supposed to supervise, but I am not do not know why, but I am not.?surprised.
To your question: SHE is wrong, really wrong!
Ren? J.
?
--- On Wed, 10/5/11, Jenny Vega <histotech411 <@t> gmail.com> wrote:
From: Jenny Vega <histotech411 <@t> gmail.com>
Subject: [Histonet] Difference between Neutral Buffered Formalin 10% and NB formaldehyde 3.7 to 4%
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, October 5, 2011, 7:03 PM
Ok I want to know the diferrence between Neutral Buffered Formalin 10% and NB formaldehyde 3.7 to 4% . I was taugh at college that they were the same thing and in the book from Frieda Carson it says that but my supervisor swears they are different chemicals. In the laboratory she has used 10% neutral buffered formalin all the time, but when we ran out of it? we were sent NB formaldehyde 3.7 to 4 %she sent it back and we were sent NB formaldehyde 3.7 to 4% once again. She says that 10% NBF is more concentrated and pure than 3.7 to 4 % formaldehyde and that the tissues are going to? putrefied if they are put in that solution.
Am I wrong or is she wrong? thanks
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------------------------------
Message: 4
Date: Thu, 06 Oct 2011 10:31:53 -0400
From: "Angela Bitting" <akbitting <@t> geisinger.edu>
Subject: Re: [Histonet] (no subject)
To: "Janice Mahoney" <mamawooo <@t> hotmail.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4E8D8398.2B7F.00C9.1 <@t> geisinger.edu>
Content-Type: text/plain; charset="us-ascii"
I too spoke to one vendor who claimed that the floaters are not an issue and that their bottles drew below the level of the "floating" floaters. I didn't feel insulted, but did feel that this salesperson obviously had no actual lab experience or they never would have made such a ridiculous claim.
I always put the opinions and experiences of other Histotechs far ahead of anything I hear from sales reps.
Angela Bitting, HT(ASCP), QIHC
Technical Specialist, Histology
Geisinger Medical Center
100 N Academy Ave. MC 23-00
Danville, PA 17822
phone 570-214-9634
fax 570-271-5916
>>> Janice Mahoney <mamawooo <@t> hotmail.com> 10/5/2011 8:11 PM >>>
Hello everyone,After being home from the NSH for a few weeks I have been pondering an issue that I think bears discussion on the histonet.There have been several papers published regarding "floaters" and the amount determined to come from traditional staining buckets. There was also a poster presented at the NSH this year on the subject.When I approached several vendors of H&E stainers about this issue. The answers were surprisingly pretty much the same. It is not an issue! Now I understand how one company can make this claim as their stainer uses fresh stain on each slide. The explanations from the other companies were insulting and just plain did not make sense to me. I was told by a Histo tech vendor that "All Histo techs know that floaters come from the water bath." Well, she was talking to a histo tech and I know for a fact that floaters come from a variety of places. I have seen them from the doctor's office or procedure room to the stainer and every step in be
tween. Sometimes if the "floater" is in the block it is very difficult to determine where it originated. We can however eliminate the water bath and stainer as the origin in these cases. One company told me that the design of the solution bottle eliminated floaters because floaters float and their stainer draws solutions from the bottom of the bottle. I have probably changed thousands of staining dishes during my 40+ year career (yes, I am old) and I have seen lots of little pieces of tissue at the bottom of the staining dishes. So, no, not all floaters float. I would love to hear feedback from others on this. I guess I would appreciate feedback about the floater issue as well as how a few vendors can make such claims and expect Histology techs to buy it. I really felt that a few comments were insulting to our profession and to the knowledge and expertise we possess. JanOmaha _______________________________________________
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FN:Bitting, Angela
TEL;WORK:570-271-6844
ORG:;Histology
EMAIL;WORK;PREF;NGW:AKBITTING <@t> geisinger.edu
N:Bitting;Angela
END:VCARD
------------------------------
Message: 5
Date: Thu, 6 Oct 2011 08:56:55 -0600
From: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>
Subject: RE: [Histonet] Difference between Neutral Buffered Formalin
10% andNBformaldehyde 3.7 to 4%
To: "Erikstad, Gudrun Hovstein" <Gudrun.Erikstad <@t> stolav.no>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4D14F0FC9316DD41972D5F03C070908B051DFB21 <@t> nmdamailsvr.nmda.ad.nmsu.edu>
Content-Type: text/plain; charset="us-ascii"
Now, wait just a minute! Don't tell me that after 40+ years of being a
histotech that I've been WRONG all this time??? FORMALDEHYDE is
FORMALDEHYDE is FORMALDEHYDE. FORMALIN is the diluted form of
FORMALDEHYDE, buffered and used at (usually) 10%. If I'm wrong, this
will be my last day at work... :)
------------------------------
Message: 6
Date: Thu, 6 Oct 2011 11:01:12 -0400
From: Ken_Marissael <@t> vwr.com
Subject: [Histonet] Ken Marissael is out of the office
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF8ABF2342.81253D17-ON85257921.00528237-85257921.00528237 <@t> vwr.com>
Content-Type: text/plain; charset=US-ASCII
I will be out of the office starting 10/06/2011 and will not return until
10/12/2011.
I will be away on 10/06 and return 10/12. I will have limited e-mail and
cell phone access, but will try to get back to you as quickly as possible.
While I am away, please contact VWR Healthcare Customer Service at
877-881-1192. If you require immediate attenion, please contact Jackie
Zerillo at 609-410-6152. She is the NYC rep.
------------------------------
Message: 7
Date: Thu, 6 Oct 2011 08:19:25 -0700
From: "Morken, Timothy" <Timothy.Morken <@t> ucsfmedctr.org>
Subject: RE: [Histonet] Floaters
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<8D7C2D242DBD45498006B21122072BF89448A27E <@t> MCINFRWEM003.ucsfmedicalcenter.org>
Content-Type: text/plain; charset=iso-8859-1
The recent paper by Platt et all (Tissue Floaters and Contaminants in the Histology Laboratory, Arch Pathol Lab Med, Vol 133, June 2009) details floaters from waterbaths and staining process pretty well.
Waterbaths do not appear to be a contributor. They literally found only ONE tissue fragment in waterbath water saved from an entire day of sectioning from 13 waterbaths. And it was in the water, not on a slide. They apparently did not identify ANY waterbath-derived floaters on any slide. They attribute this finding to the fact the tissue is still in paraffin so the tissue was either on the slide or stuck to the side of the waterbath, not floating in the water.
To determine the extent of staining-derived floaters they measured the amount of tissue fragments found in each staining station. The vast majority were in the first xylene and alcohol baths. After the 95% alcohol there were only very rare tissue fragments in the rest of the baths.
They found the staining process does contribute floaters (besides those that are clearly in the block), but apparently the do not derive from the fragments in bottom of the reagent containers. The floaters were from the other slides in the specific rack stained. The key finding was that BLANK slides put through a stainer at the end of the day (after staining over 1300 slides and accumulating the maximum amount of tissue fragments in the reagent baths) DID NOT PICK UP ANY FLOATERS.
ALL the floaters that they determined had come from the staining process occurred ONLY when tissue slides were included in the stain rack (They alternated blank and tissue slides in the rack). So, the tissue seen in the stain dish does not appear to contribute to floaters.
A stain instrument vendor repeated this experiment in our lab and came up with the same result.
Our pathologists were mixed on the floater question. Some felt a system that eliminated floaters would be beneficial. Others thought floaters were easily detected and the vast majority could be ignored. It remains to be determined it is cost effective to replace the current system with one that would eliminate floaters. It does not appear to be a sole reason to replace the current system.
Tim Morken
Supervisor, Histology, IPOX
UCSF Medical Center
San Francisco, CA, USA
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, October 06, 2011 6:34 AM
To: histonet <@t> lists.utsouthwestern.edu; Janice Mahoney
Subject: [Histonet] Floaters
This is a very interesting subject indeed. To start I always take "with a grain of salt" whatever a vendor tells me because their opinions may (or may not, but just in case) biased to the products they sell. The same thing goes for any automobiles salesperson of any automobile make claiming the "wonders" of the make they sell compared with other makes.
Now, to the floater. I have also seen pieces of sections on the bottom of staining dishes. They are there because they fell-off the stained section. My question is: what are the possibilities of that piece of lost stained section that fell from its original slide, "re-float" and magically adhere to another slide? I really think that the possibilities range from almost none, to none.
Going to the water bath: yes, this is a major source of floaters because a piece of a section left in the water bath can be picked up in another slide along with a section from another case. That is why it is mandatory to swipe the surface of the water bath with? a tissue after each case is cut. The source can be defined if we keep a record of the sequence in which the blocks are cut (as was standard procedure at my lab).
Whenever we ended with a floater in a section we went immediately to the block to determine if the floater came from the block or the water bath.
The other sources, either during description, cassetting or embedding are going to end in the block and it is very difficult to determine the origin. The only thing to do is to make sure that embedding wells where the forceps rest and heat, are thoroughly cleaned when the embedding ends.
But I think that the main thrust of the question deals with floaters coming from the staining dishes: as I wrote earlier I give to this source?a very slim and almost impossible chance.
Ren? J.
?
?
?
--- On Wed, 10/5/11, Janice Mahoney <mamawooo <@t> hotmail.com> wrote:
From: Janice Mahoney <mamawooo <@t> hotmail.com>
Subject: [Histonet] (no subject)
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, October 5, 2011, 8:11 PM
Hello everyone,After being home from the NSH for a few weeks I have been pondering an issue that I think bears discussion on the histonet.There have been several papers published regarding "floaters" and the amount determined to come from traditional staining buckets.? There was also a poster presented at the NSH this year on the subject.When I approached several vendors of H&E stainers about this issue.? The answers were surprisingly pretty much the same.???It is not an issue!? Now I understand how one company can make this claim as their stainer uses fresh stain on each slide.? The explanations from the other companies were insulting and just plain did not make sense to me.? I was told by a Histo tech vendor that "All Histo techs know that floaters come from the water bath."? Well, she was talking to a histo tech and I know for a fact that floaters come from a variety of places.? I have seen them from the doctor's office or procedure room to
the stainer and every step in between.? Sometimes if the "floater" is in the block it is very difficult to determine where it originated.? We can however eliminate the water bath and stainer as the origin in these cases.? One company told me that the design of the solution bottle eliminated floaters because floaters float and their stainer draws solutions from the bottom of the bottle.? I have probably changed thousands of staining dishes during my 40+ year career (yes, I am old) and I have seen lots of little pieces of tissue at the bottom of the staining dishes.? So, no, not all floaters float.? I would love to hear feedback from others on this.? I guess I would appreciate feedback about the floater issue as well as how a few vendors can make such claims and expect Histology techs to buy it.? I really felt that a few comments were insulting to our profession and to the knowledge and expertise we possess. JanOmaha ??? ????????
?????? ??? ? _______________________________________________
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------------------------------
Message: 8
Date: Thu, 6 Oct 2011 05:23:53 -1000
From: Janice Mahoney <mamawooo <@t> hotmail.com>
Subject: RE: [Histonet] (no subject)
To: Cindy Robinson <robinsoc <@t> mercyhealth.com>, histo net
<histonet <@t> lists.utsouthwestern.edu>, Christie Gowan
<christiegowan <@t> msn.com>
Message-ID: <BLU159-W54FCAC94954BE33D0EE15BD7F90 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
Very good question Cindi.Jan
> Date: Thu, 6 Oct 2011 09:26:18 -0400
> From: robinsoc <@t> mercyhealth.com
> To: mamawooo <@t> hotmail.com; histonet <@t> lists.utsouthwestern.edu; christiegowan <@t> msn.com
> Subject: RE: [Histonet] (no subject)
>
> It is a good thing this vendor does not work in a histo lab because with that comment/attitude he/she would not last long. I have heard the water bath theory but upon investigation this has never been the source because every histotech I work with cleans it each and every time. We have traced floaters back to grossing stations, processors...especially placenta getting 'snagged' on a piece of bone...embedding forceps (we now use ones with no grooves), forcep warmers, stains and coverslippers. We have significantly reduced the number we see since we make the effort to track each one found as an ongoing QA project, but we still see the occasional floater. We did find that we have to put tissue types which fragment easily into mesh cassettes or bags but this can cause issues with fluid exchange and carryover during processing. It is a balancing act. I have always wondering about the newer processors with orientation cassettes which are embed and then cut without opening. Do th
ey see less floaters with this type of 'closed' system?
>
>
>
> Cindi Robinson HT(ASCP)
> Mercy Medical Center
> Dunes Medical Laboratories
> 350 W Anchor Dr
> Dakota Dunes SD 57049
> phone-712-279-2768
> robinsoc <@t> mercyhealth.com
>
>
> >>> CHRISTIE GOWAN <christiegowan <@t> msn.com> 10/6/2011 7:44 AM >>>
>
> I agree with you Jan. In my 30 + years as a histotech (yes, I am old too) I have seen floaters come from a variety of places but I am hard pressed to remember any floater coming from the water bath. Today we are blessed with DNA fingerprinting to determine if the floater is or is not from the patient but that still does not address the real issue of where did it come from and how do we stop it. The vendors stating that it is not an issue have never been re-biopsied because of a floater in with their tissue. Good discussion and long overdue. I look forward to the day when it is no longer an automatic response from all involved that it is "Histology's fault". Hope you are enjoying your new adventures in retirement. See you next year in Canada!
>
>
> > From: mamawooo <@t> hotmail.com
> > To: histonet <@t> lists.utsouthwestern.edu
> > Date: Wed, 5 Oct 2011 14:11:25 -1000
> > Subject: [Histonet] (no subject)
> >
> >
> > Hello everyone,After being home from the NSH for a few weeks I have been pondering an issue that I think bears discussion on the histonet.There have been several papers published regarding "floaters" and the amount determined to come from traditional staining buckets. There was also a poster presented at the NSH this year on the subject.When I approached several vendors of H&E stainers about this issue. The answers were surprisingly pretty much the same. It is not an issue! Now I understand how one company can make this claim as their stainer uses fresh stain on each slide. The explanations from the other companies were insulting and just plain did not make sense to me. I was told by a Histo tech vendor that "All Histo techs know that floaters come from the water bath." Well, she was talking to a histo tech and I know for a fact that floaters come from a variety of places. I have seen them from the doctor's office or procedure room to the stainer and every step in between
. Sometimes if the "floater" is in the block it is very difficult to determine where it originated. We can however eliminate the water bath and stainer as the origin in these cases. One company told me that the design of the solution bottle eliminated floaters because floaters float and their stainer draws solutions from the bottom of the bottle. I have probably changed thousands of staining dishes during my 40+ year career (yes, I am old) and I have seen lots of little pieces of tissue at the bottom of the staining dishes. So, no, not all floaters float. I would love to hear feedback from others on this. I guess I would appreciate feedback about the floater issue as well as how a few vendors can make such claims and expect Histology techs to buy it. I really felt that a few comments were insulting to our profession and to the knowledge and expertise we possess. JanOmaha _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> _______________________________________________
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------------------------------
Message: 9
Date: Thu, 6 Oct 2011 17:37:57 +0200
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: [Histonet] "formol"
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4C8F235CA0384093830B90BB552641EA <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
Formol is an old brandname, I think for a desinfection reagent. Not really
custom any more.
Bye (another) Gudrun
-----Urspr?ngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Rene J
Buesa
Gesendet: Donnerstag, 06. Oktober 2011 15:13
An: histonet <@t> lists.utsouthwestern.edu; Jenny Vega
Betreff: [Histonet] Difference between Neutral Buffered Formalin 10% and
NBformaldehyde 3.7 to 4%
Jenny:
Formaldehyde is?the GAS methanal?that dissolves in water at a maximum
concentration between 37-40% vol./vol. To prevent its polymerization, small
amounts of mathanol (an alcohol)are added.
The 10% solution formaldehyde contains between 3.7 and 4.0% of formaldehyde
and is called "formalin" in many countries and "formol" in Europe.
So formalin is a 3.7 to 4.0% solution of formaldehyde.
So a 10% formalin = 3.7-4.0% formaldehyde.
NBF is prepared by added phosphate salts to prevent the formalin to acidify
and is used to fix tissues.
It is a pitty that your supervisor has so little knowledge about the area
she is supposed to supervise, but I am not do not know why, but I am
not.?surprised.
To your question: SHE is wrong, really wrong!
Ren? J.
?
--- On Wed, 10/5/11, Jenny Vega <histotech411 <@t> gmail.com> wrote:
From: Jenny Vega <histotech411 <@t> gmail.com>
Subject: [Histonet] Difference between Neutral Buffered Formalin 10% and NB
formaldehyde 3.7 to 4%
To: histonet <@t> lists.utsouthwestern.edu
Date: Wednesday, October 5, 2011, 7:03 PM
Ok I want to know the diferrence between Neutral Buffered Formalin 10% and
NB formaldehyde 3.7 to 4% . I was taugh at college that they were the same
thing and in the book from Frieda Carson it says that but my supervisor
swears they are different chemicals. In the laboratory she has used 10%
neutral buffered formalin all the time, but when we ran out of it? we were
sent NB formaldehyde 3.7 to 4 %she sent it back and we were sent NB
formaldehyde 3.7 to 4% once again. She says that 10% NBF is more
concentrated and pure than 3.7 to 4 % formaldehyde and that the tissues are
going to? putrefied if they are put in that solution.
Am I wrong or is she wrong? thanks
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------------------------------
Message: 10
Date: Thu, 6 Oct 2011 15:43:15 +0000
From: Kelly Cross <KCross <@t> cvm.tamu.edu>
Subject: [Histonet] control for Hemoglobin?
To: "Histonet Listserv (E-mail) (histonet <@t> lists.utsouthwestern.edu)"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<869848DDBB7C5D4896A569A38B814E690551CAD1 <@t> CVMMB01.cvm.tamu.edu>
Content-Type: text/plain; charset="us-ascii"
Hi Histonetters,
What control should I use for Okajima's Stain for hemoglobin?
Thank you for your help!
Kelly Cross B.S., HT (ASCP)
Medical Laboratory Supervisor
Texas A&M University
VTPB Histology Lab VMA bldg. 214
College Station, TX 77843-4467
Office: 979-862-3658
Lab: 979-845-5149
------------------------------
Message: 11
Date: Thu, 6 Oct 2011 05:44:31 -1000
From: Janice Mahoney <mamawooo <@t> hotmail.com>
Subject: RE: [Histonet] Tissue left in processor
To: <rjbuesa <@t> yahoo.com>, histo net
<histonet <@t> lists.utsouthwestern.edu>, <rcharles <@t> pa.gov>
Message-ID: <BLU159-W19BD271DE85D9341C51DD7D7F90 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
I agree with Rene, as long as the temp is only a few degrees above the melting point of the paraffin.Jan,Omaha
> Date: Thu, 6 Oct 2011 07:13:54 -0700
> From: rjbuesa <@t> yahoo.com
> To: histonet <@t> lists.utsouthwestern.edu; rcharles <@t> pa.gov
> Subject: Re: [Histonet] Tissue left in processor
> CC:
>
> I do not think that a well fixed, well processed tissue left in molten paraffin for 2 hours after the processor finished will have any adverse outcome.
> Ren? J.
>
> --- On Thu, 10/6/11, Charles, Roger <rcharles <@t> pa.gov> wrote:
>
>
> From: Charles, Roger <rcharles <@t> pa.gov>
> Subject: [Histonet] Tissue left in processor
> To: "Histonet (histonet <@t> lists.utsouthwestern.edu)" <histonet <@t> lists.utsouthwestern.edu>
> Date: Thursday, October 6, 2011, 10:05 AM
>
>
> Hi All,
> Is there any standard on how long tissue cassettes can remain in the processor after processing before the tissue is subjected to unwanted outcomes? And if so what type of artifacts can one expect from tissue that was in the processor in molten paraffin for 2 hours after the processing was completed?
> Thanks so much.
> Roger
>
> Roger Charles| Microbiologist II
> Pennsylvania Veterinary Laboratory
> 2305 North Cameron Street | Harrisburg, PA 17110
> Phone: 717.787.8808 | Fax: 717.772.3895
> www.agriculture.state.pa.us<http://www.agriculture.state.pa.us>
>
> _______________________________________________
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 12
Date: Thu, 06 Oct 2011 11:49:08 -0400
From: "Tammy Pickles" <Tammy.Pickles <@t> inspection.gc.ca>
Subject: [Histonet] Imidazole buffer
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4E8D7994.B440.007F.1 <@t> inspection.gc.ca>
Content-Type: text/plain; charset="us-ascii"
Hi,
I am wondering if anyone out there can tell me the function of Imidazole, when used in a buffer. We are currently using it to make up our in house DAB solution. The results are fantastic with an overall increase in sensitivity with all of our antibodies. The only information I can find concerning the function is to help in blocking nonspecific staining. Does anyone have more information to share?
Thanks in advance,
Tammy
------------------------------
Message: 13
Date: Thu, 6 Oct 2011 09:59:32 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: [Histonet] RE: DAPI on thicker sections, problem with
gradient
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000601cc8440$f1af4270$d50dc750$@bresnan.net>
Content-Type: text/plain; charset="iso-8859-1"
Carmen,
You wrote:
Good morning:
I am Carmen Garcia from the Faculty of Medicine of the University of
Valencia, Spain. I usually use your vectashield mounting medium for
fluorescence with DAPI (H-1200) and I never have any problem.
But now I am working with 50um frozen sections and the problem that I
have is that the DAPI staining does not arrive to the bottom of the
section. Do you have any sugesstion of what can I do? (I already have
the sections permeabilized with triton x-100) I have thougth to
incubate with the mounting medium some hours at 4?C or put some
mounting medium in the antibody diluent and incubate...
any suggestion would be really welcome.
Thank you so much,
Kind regards,
Carmen
****************************************************************************
***********
The problem you are experiencing is what we refer to as a DAPI staining
gradient. Uneven staining occurs with the DAPI is not reaching all the
cells due to viscosity of the mounting media and thickness of the tissue
section.
You did not say whether your thick sections have been fixed before
sectioning and staining for a tissue component, and then the DAPI - please
explain. I suggest you do NOT use a mounting media with DAPI, but rather
make a DAPI solution which you can then apply to the section and incubate
mounting the coverslip with Vectashield that does not contain DAPI. Using a
DAPI solution may shorten the time you now use, e.g. "some hours". You
can buy the DAPI, ready to use from Biogenex or other vendors, but making it
in house is simple. I would assume you could add Triton X to the DAPI
solution to improve permeability and match what you have already done with
that detergent. After DAPI staining, mount the coverslip using VectaShield
without DAPI. DAPI recipes can be found on the internet, just Google DAPI.
I found it on a Johns Hopkins webpage, but IHC world may have it too.
There are other sources of ready to use DAPI if you look.
We have used Prolong Gold antifade reagent with DAPI, and get a staining
gradient on 5 um sections. This is very annoying, and probably caused by
the actual technique of cover slipping itself. We are now going to use
Biogenex ready to use DAPI before the Prolong Gold antifade reagent without
DAPUI. Staining for a thin section takes 5 mintues at RT, so you should
try several 50 um sections (non experimental) to determine your optimal
staining time. It may take only 15 minutes although cold, 4C temperature
may slow down the penetration of the stain through the thicker section.
This way you can achieve even staining of your nuclei without long
incubations in the more viscious mounting media containing DAPI.
Good luck, and let us know if you have success...................
Gayle M. Callis
HTL/HT/MT(ASCP)
------------------------------
Message: 14
Date: Thu, 6 Oct 2011 16:06:06 +0000
From: Helen Fedor <hfedor <@t> jhmi.edu>
Subject: RE: [Histonet] RE: DAPI on thicker sections, problem with
gradient
To: 'gayle callis' <gayle.callis <@t> bresnan.net>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<BCE07FB3A86FD040969A9EC0DD3EDA0D0B87937F <@t> JHEMTMWEX4.win.ad.jhu.edu>
Content-Type: text/plain; charset="iso-8859-1"
Hello, Maybe this method was meant to be used on Floating sections instead of mounted sections. That way the binding and penetration would go from both sides of the tissue.
Helen L. Fedor
http://tmalab.jhmi.edu/
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of gayle callis
Sent: Thursday, October 06, 2011 12:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: DAPI on thicker sections, problem with gradient
Carmen,
You wrote:
Good morning:
I am Carmen Garcia from the Faculty of Medicine of the University of
Valencia, Spain. I usually use your vectashield mounting medium for
fluorescence with DAPI (H-1200) and I never have any problem.
But now I am working with 50um frozen sections and the problem that I
have is that the DAPI staining does not arrive to the bottom of the
section. Do you have any sugesstion of what can I do? (I already have
the sections permeabilized with triton x-100) I have thougth to
incubate with the mounting medium some hours at 4?C or put some
mounting medium in the antibody diluent and incubate...
any suggestion would be really welcome.
Thank you so much,
Kind regards,
Carmen
****************************************************************************
***********
The problem you are experiencing is what we refer to as a DAPI staining
gradient. Uneven staining occurs with the DAPI is not reaching all the
cells due to viscosity of the mounting media and thickness of the tissue
section.
You did not say whether your thick sections have been fixed before
sectioning and staining for a tissue component, and then the DAPI - please
explain. I suggest you do NOT use a mounting media with DAPI, but rather
make a DAPI solution which you can then apply to the section and incubate
mounting the coverslip with Vectashield that does not contain DAPI. Using a
DAPI solution may shorten the time you now use, e.g. "some hours". You
can buy the DAPI, ready to use from Biogenex or other vendors, but making it
in house is simple. I would assume you could add Triton X to the DAPI
solution to improve permeability and match what you have already done with
that detergent. After DAPI staining, mount the coverslip using VectaShield
without DAPI. DAPI recipes can be found on the internet, just Google DAPI.
I found it on a Johns Hopkins webpage, but IHC world may have it too.
There are other sources of ready to use DAPI if you look.
We have used Prolong Gold antifade reagent with DAPI, and get a staining
gradient on 5 um sections. This is very annoying, and probably caused by
the actual technique of cover slipping itself. We are now going to use
Biogenex ready to use DAPI before the Prolong Gold antifade reagent without
DAPUI. Staining for a thin section takes 5 mintues at RT, so you should
try several 50 um sections (non experimental) to determine your optimal
staining time. It may take only 15 minutes although cold, 4C temperature
may slow down the penetration of the stain through the thicker section.
This way you can achieve even staining of your nuclei without long
incubations in the more viscious mounting media containing DAPI.
Good luck, and let us know if you have success...................
Gayle M. Callis
HTL/HT/MT(ASCP)
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 15
Date: Thu, 6 Oct 2011 16:09:28 +0000
From: <idimitro <@t> mun.ca>
Subject: [Histonet] RE: Cutting frozen sections from visceral fat
To: <sdysart <@t> mirnarx.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<14C3108E8B98EF43B3EDAE58336700340511A2 <@t> exchange.med.mun.ca>
Content-Type: text/plain; charset="us-ascii"
Hi Sarah,
Thank you for the response. We are a research lab so whatever the researchers want to be done we do it with all kinds of tissues. In this case they are looking at the size of the fat cells in the animals.
I will try to lower the temperature to -30 as was also suggested to me and if this does not work I will look into fixation. In Bancroft's fifth edition is mentioned that if you need fixation you need to use formol-calcium. I hope this will harden the fat and stabilize the membrane proteins enough so I can cut it. I will let you know how it worked.
Thanks again,
iliana
-----Original Message-----
From: Sarah Dysart [mailto:sdysart <@t> mirnarx.com]
Sent: October 6, 2011 12:14 PM
To: Dimitrova, Iliana
Subject: RE: Cutting frozen sections from visceral fat
Cutting frozen fat is basically impossible. It will just goo up and rip out of the chuck of frozen OCT. I think there is a way where you can do a fixation process first in something, but I'm not sure of that. Why do they need frozen fat sections? Cutting fat after fixation is totally doable with the right fixation. I have done this when people have asked for frozen fat sections. I can usually talk them out of the frozen and just do the FFPE sections. Let me know I can give you more details.
http://www.pathologyinnovations.com/new_page_2.htm
This site has a section in it called wrestling the fat one, that has some ideas for you.
Good Luck!!
Sarah Goebel-Dysart, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas 78744
(512)901-0900 ext. 6912
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of idimitro <@t> mun.ca
Sent: Thursday, October 06, 2011 6:59 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Cutting frozen sections from visceral fat
Hi everyone,
I am having trouble cutting frozen sections from visceral fat. I have never cut only fat before. I am also cutting liver sections and they are perfect, but when I try to cut the fat it is not working.
The temperature of the cryostat is ( -20) degree Celsius.
I wonder if anyone has ever done this. I was thinking that if I lower the temperature or fix the fat with formol-calcium I may have success.
Iliana Dimitrova, MSc, RT
Histology Supervisor
Room 2808
Medical Education and Laboratory Support Services (MELSS)
Faculty of Medicine
Memorial University
St. John's, NL
This electronic communication is governed by the terms and conditions at
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------------------------------
Message: 16
Date: Thu, 6 Oct 2011 12:30:02 -0400
From: "Dessoye, Michael J" <mjdessoye <@t> wvhcs.org>
Subject: [Histonet] CMV/HSV Protocols
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E2547E1CD0EE324488A2940994571EFA04548EDA <@t> WVHCS-EXCHANGE.wvhcs.com>
Content-Type: text/plain; charset="us-ascii"
Does anyone have protocols for CMV, HSV I, and HSV II (from Cell Marque)
that they would be willing to share for the Benchmark Ultra (i-View)?
Thanks!
Michael J. Dessoye, M.S. | Histology Supervisor | Wyoming Valley Health
Care System | mjdessoye <@t> wvhcs.org <mailto:mjdessoye <@t> wvhcs.org> |
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1485 | Fax:
570-552-1526
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------------------------------
Message: 17
Date: Thu, 6 Oct 2011 16:39:38 +0000
From: joelle weaver <joelleweaver <@t> hotmail.com>
Subject: RE: [Histonet] Floaters
To: <timothy.morken <@t> ucsfmedctr.org>, Histonet
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <SNT135-W459933889C3BBE36037729D8F90 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
This was an excellent article, I archived it, interesting that your own experiments confirmed this results.
Joelle Weaver MAOM, BA, (HTL) ASCP
> From: Timothy.Morken <@t> ucsfmedctr.org
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Thu, 6 Oct 2011 08:19:25 -0700
> Subject: RE: [Histonet] Floaters
>
> The recent paper by Platt et all (Tissue Floaters and Contaminants in the Histology Laboratory, Arch Pathol Lab Med, Vol 133, June 2009) details floaters from waterbaths and staining process pretty well.
>
> Waterbaths do not appear to be a contributor. They literally found only ONE tissue fragment in waterbath water saved from an entire day of sectioning from 13 waterbaths. And it was in the water, not on a slide. They apparently did not identify ANY waterbath-derived floaters on any slide. They attribute this finding to the fact the tissue is still in paraffin so the tissue was either on the slide or stuck to the side of the waterbath, not floating in the water.
>
> To determine the extent of staining-derived floaters they measured the amount of tissue fragments found in each staining station. The vast majority were in the first xylene and alcohol baths. After the 95% alcohol there were only very rare tissue fragments in the rest of the baths.
>
> They found the staining process does contribute floaters (besides those that are clearly in the block), but apparently the do not derive from the fragments in bottom of the reagent containers. The floaters were from the other slides in the specific rack stained. The key finding was that BLANK slides put through a stainer at the end of the day (after staining over 1300 slides and accumulating the maximum amount of tissue fragments in the reagent baths) DID NOT PICK UP ANY FLOATERS.
>
> ALL the floaters that they determined had come from the staining process occurred ONLY when tissue slides were included in the stain rack (They alternated blank and tissue slides in the rack). So, the tissue seen in the stain dish does not appear to contribute to floaters.
>
> A stain instrument vendor repeated this experiment in our lab and came up with the same result.
>
> Our pathologists were mixed on the floater question. Some felt a system that eliminated floaters would be beneficial. Others thought floaters were easily detected and the vast majority could be ignored. It remains to be determined it is cost effective to replace the current system with one that would eliminate floaters. It does not appear to be a sole reason to replace the current system.
>
> Tim Morken
> Supervisor, Histology, IPOX
> UCSF Medical Center
> San Francisco, CA, USA
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
> Sent: Thursday, October 06, 2011 6:34 AM
> To: histonet <@t> lists.utsouthwestern.edu; Janice Mahoney
> Subject: [Histonet] Floaters
>
> This is a very interesting subject indeed. To start I always take "with a grain of salt" whatever a vendor tells me because their opinions may (or may not, but just in case) biased to the products they sell. The same thing goes for any automobiles salesperson of any automobile make claiming the "wonders" of the make they sell compared with other makes.
> Now, to the floater. I have also seen pieces of sections on the bottom of staining dishes. They are there because they fell-off the stained section. My question is: what are the possibilities of that piece of lost stained section that fell from its original slide, "re-float" and magically adhere to another slide? I really think that the possibilities range from almost none, to none.
> Going to the water bath: yes, this is a major source of floaters because a piece of a section left in the water bath can be picked up in another slide along with a section from another case. That is why it is mandatory to swipe the surface of the water bath with a tissue after each case is cut. The source can be defined if we keep a record of the sequence in which the blocks are cut (as was standard procedure at my lab).
> Whenever we ended with a floater in a section we went immediately to the block to determine if the floater came from the block or the water bath.
> The other sources, either during description, cassetting or embedding are going to end in the block and it is very difficult to determine the origin. The only thing to do is to make sure that embedding wells where the forceps rest and heat, are thoroughly cleaned when the embedding ends.
> But I think that the main thrust of the question deals with floaters coming from the staining dishes: as I wrote earlier I give to this source a very slim and almost impossible chance.
> Ren? J.
>
>
>
>
> --- On Wed, 10/5/11, Janice Mahoney <mamawooo <@t> hotmail.com> wrote:
>
>
> From: Janice Mahoney <mamawooo <@t> hotmail.com>
> Subject: [Histonet] (no subject)
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Wednesday, October 5, 2011, 8:11 PM
>
>
>
> Hello everyone,After being home from the NSH for a few weeks I have been pondering an issue that I think bears discussion on the histonet.There have been several papers published regarding "floaters" and the amount determined to come from traditional staining buckets. There was also a poster presented at the NSH this year on the subject.When I approached several vendors of H&E stainers about this issue. The answers were surprisingly pretty much the same. It is not an issue! Now I understand how one company can make this claim as their stainer uses fresh stain on each slide. The explanations from the other companies were insulting and just plain did not make sense to me. I was told by a Histo tech vendor that "All Histo techs know that floaters come from the water bath." Well, she was talking to a histo tech and I know for a fact that floaters come from a variety of places. I have seen them from the doctor's office or procedure room to
> the stainer and every step in between. Sometimes if the "floater" is in the block it is very difficult to determine where it originated. We can however eliminate the water bath and stainer as the origin in these cases. One company told me that the design of the solution bottle eliminated floaters because floaters float and their stainer draws solutions from the bottom of the bottle. I have probably changed thousands of staining dishes during my 40+ year career (yes, I am old) and I have seen lots of little pieces of tissue at the bottom of the staining dishes. So, no, not all floaters float. I would love to hear feedback from others on this. I guess I would appreciate feedback about the floater issue as well as how a few vendors can make such claims and expect Histology techs to buy it. I really felt that a few comments were insulting to our profession and to the knowledge and expertise we possess. JanOmaha
> _______________________________________________
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
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------------------------------
Message: 18
Date: Thu, 6 Oct 2011 16:55:09 +0000
From: joelle weaver <joelleweaver <@t> hotmail.com>
Subject: RE: [Histonet] Difference between Neutral Buffered Formalin
10% andNBformaldehyde 3.7 to 4%
To: <sbreeden <@t> nmda.nmsu.edu>, <gudrun.erikstad <@t> stolav.no>, Histonet
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <SNT135-W378FBA45C4CC8C65E92FDAD8F90 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
I like this article's discussion of the history of usuage and terminology: citation Fox, C., Johnson, F., Whiting, J., and Roller, P. Formaldehyde Fixation, The Journal of Histochemistry and Cytochemistry, Vol. 33, No.8, pp. 845-853, 1985.
Joelle Weaver MAOM, BA, (HTL) ASCP
> Date: Thu, 6 Oct 2011 08:56:55 -0600
> From: sbreeden <@t> nmda.nmsu.edu
> To: Gudrun.Erikstad <@t> stolav.no; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] Difference between Neutral Buffered Formalin 10% andNBformaldehyde 3.7 to 4%
> CC:
>
> Now, wait just a minute! Don't tell me that after 40+ years of being a
> histotech that I've been WRONG all this time??? FORMALDEHYDE is
> FORMALDEHYDE is FORMALDEHYDE. FORMALIN is the diluted form of
> FORMALDEHYDE, buffered and used at (usually) 10%. If I'm wrong, this
> will be my last day at work... :)
>
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 19
Date: Thu, 6 Oct 2011 10:55:11 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: RE: [Histonet] Tissue left in processor
To: "'Janice Mahoney'" <mamawooo <@t> hotmail.com>, <rjbuesa <@t> yahoo.com>,
"'histo net'" <histonet <@t> lists.utsouthwestern.edu>, <rcharles <@t> pa.gov>
Message-ID: <000001cc8448$b806d8f0$28148ad0$@bresnan.net>
Content-Type: text/plain; charset="iso-8859-1"
Some prefer to take the cassettes off the processor and let them harden
until embedding can be done, rather than leave in molten, hot paraffin.
This does not damage the tissue, and placing the cooled cassettes into the
embedding center holding area allows the paraffin to re-melt in a shorter
time before embedding. This has been discussed in the past on Histonet.
Some tissues may dry out even more WITH difficult sectioning after prolonged
heat exposure. One result will be a parched earth effect seen in sections.
I see you are from a veterinary facility, and if you work with rodent
tissues e.g. spleen and liver, you should not let the tissue sit in paraffin
or you will have little hard rocks to section. If your sectioning suffers
(dry, friable, shattered, hard) after allowing tissues to sit in hot
paraffin as compared to the times when you embed asap after processing is
finished, then over exposure to hot paraffin can contributing factor.
I personally do not like to "cook" my tissue any longer than necessary and
heat labile antigens will also be at risk. I schedule so I can be there to
embed when the processing is completed.
Our standard is to embed when processing is finished and schedule
accordingly.
Gayle Callis
HTL/HT/MT(ASCP)
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Janice
Mahoney
Sent: Thursday, October 06, 2011 9:45 AM
To: rjbuesa <@t> yahoo.com; histo net; rcharles <@t> pa.gov
Subject: RE: [Histonet] Tissue left in processor
I agree with Rene, as long as the temp is only a few degrees above the
melting point of the paraffin.Jan,Omaha
> Date: Thu, 6 Oct 2011 07:13:54 -0700
> From: rjbuesa <@t> yahoo.com
> To: histonet <@t> lists.utsouthwestern.edu; rcharles <@t> pa.gov
> Subject: Re: [Histonet] Tissue left in processor
> CC:
>
> I do not think that a well fixed, well processed tissue left in molten
paraffin for 2 hours after the processor finished will have any adverse
outcome.
> Ren? J.
>
> --- On Thu, 10/6/11, Charles, Roger <rcharles <@t> pa.gov> wrote:
>
>
> From: Charles, Roger <rcharles <@t> pa.gov>
> Subject: [Histonet] Tissue left in processor
> To: "Histonet (histonet <@t> lists.utsouthwestern.edu)"
> <histonet <@t> lists.utsouthwestern.edu>
> Date: Thursday, October 6, 2011, 10:05 AM
>
>
> Hi All,
> Is there any standard on how long tissue cassettes can remain in the
processor after processing before the tissue is subjected to unwanted
outcomes? And if so what type of artifacts can one expect from tissue that
was in the processor in molten paraffin for 2 hours after the processing was
completed?
> Thanks so much.
> Roger
>
> Roger Charles| Microbiologist II
> Pennsylvania Veterinary Laboratory
> 2305 North Cameron Street | Harrisburg, PA 17110
> Phone: 717.787.8808 | Fax: 717.772.3895
> www.agriculture.state.pa.us<http://www.agriculture.state.pa.us>
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