Re: [Histonet] Tissue Fixation

[Tony Henwood (SCHN) via Histonet]

If the specimen is placed on absorbent paper prior to fixation, the paper will 
gradually absorb the water from the specimen causing it to dry out.
Microscopically the tissue will look  a little like what you see when a laser 
scalpel is used to excise a skin specimen. The heat affected eosin stained 
collagen will look quite brilliant (even fluorescent like) and if you do a 
Masson stain for connective tissue will give a red colouration, rather than a 
green or blue you would normally see with well-fixed collagen.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: T H via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, 31 March 2017 11:51 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Fixation

Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] Tissue fixation

[Bob Richmond via Histonet]

Tim describes a problem "I have a pathologist that is not happy with the
fixation on some of our LEEP specimens."

LEEP specimens are inherently crappy, because of the cautery used to obtain
them, and the resulting cautery artifact in the specimens. In the last few
years they've turned the voltage down, and that's helped some - and perhaps
this particular clinic hasn't turned the voltage down.

I wonder how familiar your pathologist is with the whole process - the sort
of homely detail not taught in pathology residency, and the sort of detail
I often go out and research on my own. It's helpful to all to know that the
clinical stakes here are not high - we do report margins on cervical LEEP
specimens (when we can figure them out), but clinical outcomes apparently
don't have much to do with whether margins are positive for dysplasia or
not.

It's impossible to get india ink to stick to the cauterized gross margins,
but the cautery artifact is painfully obvious under the microscope anyway.

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] Tissue fixation

[Mayer,Toysha N via Histonet]

I agree with Rene.  To discredit the pathologist theory  show if all of the 
specimens from the run are not fixed properly.  Then show if it is just the 
LEEPs.  Then show if it is that particular clients specimens?  Then onto that 
client's LEEPs.
That should prove your problem lies with the client handling and not the 
fixation on your end.  Cauterization can cause burning of the specimen, but not 
look unfixed. If it was left out of formalin, autolysis can set in.
The situation shows the god-complex some physicians have and the lack of 
respect they have for their techs.  
This is why we should all be certified and keep up with continuing education, 
so that these pathologists will respect us.
Ok, I'm going to get off my soapbox now, before I say something ugly.

Toysha Mayer






--

Message: 3
Date: Fri, 31 Mar 2017 12:50:37 +
From: T H <thiggin...@msn.com>
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: [Histonet] Tissue Fixation
Message-ID:

<sn1pr19mb060735d7b896c67b0223e098d8...@sn1pr19mb0607.namprd19.prod.outlook.com>

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

Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim



--

Message: 4
Date: Fri, 31 Mar 2017 13:32:35 + (UTC)
From: Rene J Buesa <rjbu...@yahoo.com>
To: T H <thiggin...@msn.com>,   "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation
Message-ID: <1730848134.8065268.1490967155...@mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?Ren? 

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.? She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).? She claims we must be diluting our formalin to cause this 
issue or "something".? We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.? Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.? That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?? Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

___

Re: [Histonet] Tissue Fixation

[Gudrun Lang via Histonet]

I second the opinion of Joyce. We see such effects in portio-conisations, that 
are done with a thermo-electrical knife. The surface and the underlying area 
show a very pink colour in HE. It can also be seen in prostata-chips. IHC on 
such biopsies shows the effect of an non-stainable edge with a clear cut 
between positive and negative. 
https://www.uni-marburg.de/fb20/zahnerhaltk/lehre/Download/Bilder/35_plattenepithelmetaplasie_portio_02212427.jpg
I've found this picture, that shows a typical conisation-section.
regards
Gudrun 

-Ursprüngliche Nachricht-
Von: Weems, Joyce K. via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 31. März 2017 17:26
An: Morken, Timothy; Rene J Buesa; T H; Histonet@lists.utsouthwestern.edu
Betreff: Re: [Histonet] Tissue Fixation

Aren't LEEPS done with some sort of electric method that will damage the tissue 
before it even reaches formalin. I'm not positive but Google it - I believe 
that might be the problem. j

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
770-380-8099 Cell
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph’s 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.

-Original Message-
From: Morken, Timothy via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 11:07 AM
To: Rene J Buesa <rjbu...@yahoo.com>; T H <thiggin...@msn.com>
Cc: Histonet <histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation

Tim, I have to agree with Rene that the formalin or time in formalin is 
obviously not the problem - it has plenty of time in formalin (and who would 
dilute it anyway?). Handling before formalin must always be determined when 
problems arise. If the sample sits on a paper towel, gauze etc it does not 
really degrade faster, rather the tissue may dry out and so fixes by 
dehydration rather than by formalin. It may be the formalin cannot get into the 
tissue in those dried out portions of the tissue. However that is just 
speculation. Since these all seem to be from one particular client, the client 
is the place to start. The only way to determine the problem is to follow the 
specimen from start to finish. Can you or someone you trust physically observe 
the way samples are handled from the time they are taken to the time put in 
formalin? One issue I always run up against is people  saying they do one thing 
but actually doing another. And they may realize during questioning that they 
are not doing it right but don't want to admit it. It wastes a lot of time. 
I've had physicians tell me to ignore the part of the process they are 
responsible for because they do it right. I just tell them that to be complete 
we need to follow the process from start to finish, nothing personal, just 
business. Leaving any part out may lead to not resolving the problem. Probably 
99% of the process is just fine, but that 1% is damaging the sample and needs 
to be illuminated.


Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



-Original Message-
From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 6:33 AM
To: T H; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Tissue Fixation

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:


 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formali

Re: [Histonet] Tissue Fixation

[Weems, Joyce K. via Histonet]

Aren't LEEPS done with some sort of electric method that will damage the tissue 
before it even reaches formalin. I'm not positive but Google it - I believe 
that might be the problem. j

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
770-380-8099 Cell
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph’s 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.

-Original Message-
From: Morken, Timothy via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 11:07 AM
To: Rene J Buesa <rjbu...@yahoo.com>; T H <thiggin...@msn.com>
Cc: Histonet <histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation

Tim, I have to agree with Rene that the formalin or time in formalin is 
obviously not the problem - it has plenty of time in formalin (and who would 
dilute it anyway?). Handling before formalin must always be determined when 
problems arise. If the sample sits on a paper towel, gauze etc it does not 
really degrade faster, rather the tissue may dry out and so fixes by 
dehydration rather than by formalin. It may be the formalin cannot get into the 
tissue in those dried out portions of the tissue. However that is just 
speculation. Since these all seem to be from one particular client, the client 
is the place to start. The only way to determine the problem is to follow the 
specimen from start to finish. Can you or someone you trust physically observe 
the way samples are handled from the time they are taken to the time put in 
formalin? One issue I always run up against is people  saying they do one thing 
but actually doing another. And they may realize during questioning that they 
are not doing it right but don't want to admit it. It wastes a lot of time. 
I've had physicians tell me to ignore the part of the process they are 
responsible for because they do it right. I just tell them that to be complete 
we need to follow the process from start to finish, nothing personal, just 
business. Leaving any part out may lead to not resolving the problem. Probably 
99% of the process is just fine, but that 1% is damaging the sample and needs 
to be illuminated.


Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of 
Pathology UC San Francisco Medical Center



-Original Message-
From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, March 31, 2017 6:33 AM
To: T H; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Tissue Fixation

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:


 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] Tissue Fixation

[Morken, Timothy via Histonet]

Tim, I have to agree with Rene that the formalin or time in formalin is 
obviously not the problem - it has plenty of time in formalin (and who would 
dilute it anyway?). Handling before formalin must always be determined when 
problems arise. If the sample sits on a paper towel, gauze etc it does not 
really degrade faster, rather the tissue may dry out and so fixes by 
dehydration rather than by formalin. It may be the formalin cannot get into the 
tissue in those dried out portions of the tissue. However that is just 
speculation. Since these all seem to be from one particular client, the client 
is the place to start. The only way to determine the problem is to follow the 
specimen from start to finish. Can you or someone you trust physically observe 
the way samples are handled from the time they are taken to the time put in 
formalin? One issue I always run up against is people  saying they do one thing 
but actually doing another. And they may realize during questioning that they 
are not doing it right but don't want to admit it. It wastes a lot of time. 
I've had physicians tell me to ignore the part of the process they are 
responsible for because they do it right. I just tell them that to be complete 
we need to follow the process from start to finish, nothing personal, just 
business. Leaving any part out may lead to not resolving the problem. Probably 
99% of the process is just fine, but that 1% is damaging the sample and needs 
to be illuminated.


Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center



-Original Message-
From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, March 31, 2017 6:33 AM
To: T H; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Tissue Fixation

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René 

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] Tissue Fixation

[Paula Keene Pierce via Histonet]

Bravo Rene! Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, 
Inc.5830 N Blue Lake DriveNorman, OK 73069PH 405-759-3953FAX 
405-759-7513www.excaliburpathology.com

  From: Rene J Buesa via Histonet <histonet@lists.utsouthwestern.edu>
 To: T H <thiggin...@msn.com>; "histonet@lists.utsouthwestern.edu" 
<histonet@lists.utsouthwestern.edu> 
 Sent: Friday, March 31, 2017 8:44 AM
 Subject: Re: [Histonet] Tissue Fixation
   
What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René 

    On Friday, March 31, 2017 9:11 AM, T H via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] Tissue Fixation

[Rene J Buesa via Histonet]

What you describe as a possible scenario is absolutely possible.If your PT does 
not "want to hear" about it, suggest she gets a "hearing aid" or to study 
something about histotechnology or even better yet, pay attention to what a 
professional on the subject (you) has to say about it. You would never dare to 
question her diagnosis, why would she question yours on this subject?René 

On Friday, March 31, 2017 9:11 AM, T H via Histonet 
 wrote:
 

 Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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[Histonet] Tissue Fixation

[T H via Histonet]

Good Morning,


I have a pathologist that is not happy with the fixation on some of our LEEP 
specimens.  She swears its histology doing something to the specimen to cause 
the tissue to look unfixed on only "part" of the LEEP specimens (all the same 
client specimens).  She claims we must be diluting our formalin to cause this 
issue or "something".  We mentioned maybe it was on the clients end not placing 
them in 10% formalin right away, she wouldn't hear of it.


Let me give you some back ground on how our process works.  Our clients send us 
all our specimens to us via Overnight FedEx or UPS in 10% formalin they will 
then they sit in 10% formalin in-house until the processors are started around 
3pm and sits an additional 5 hours in 10% formalin on the processor before the 
processor actually starts.  That being said the fixation process has had a 
pretty good start before we ever even touch it.


My question is, and I thought I had read this in the past is, when a specimen 
is left out prior to fixation and lying on a absorbent surface such as a paper 
towel, won't the area of the tissue touching the absorbent surface begin the 
disintegration processes faster in that exact area then the rest of the 
specimen?  Or if you have any other suggestion on what might be happening to 
only "certain" specimens would be great as well.


Thanks for your help!


Tim

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Re: [Histonet] tissue fixation-formaldehyde concentrations which is best.

[Smith, Allen A]

There are many books on histological technique.  Most of them are expensive.  
The 5th edition of John Kiernan's excellent HISTOLOGICAL AND HISTOCHEMICAL 
METHODS will sell for $100 when it comes out in July.  If that is too much for 
your budget, there are older books that cover 95% of what you need to know.   
The 4th edition of Kiernan's HISTOLOGICAL AND HISTOCHEMICAL METHODS is 
available used for $60, but the price may fall when the 5th edition comes out.  
A used  4th edition of Gretchen Humason's ANIMAL TISSUE TECHNIQUES is a great 
bargain at $12.
Allen A. Smith
Professor of Anatomy
Barry University School of Podiatric Medicine
Miami Shores, Florida

-Original Message-
From: John Kiernan [mailto:jkier...@uwo.ca] 
Sent: Saturday, June 06, 2015 12:31 AM
To: Peter Noyce; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] tissue fixation-formaldehyde concentrations which is 
best.

 Dear Peter, 
 
Some of the information you mention as anecdotal is wrong. Formaldehyde and 
paraformaldehyde are well documented in original peer-reviewed papers and in 
all textbooks in the fields of histotechnology and histochemistry.  
 
Your anecdote about high concentrations of formaldehyde quickly form a 'shell' 
in the tissue and will stop good penetration and fixation to the deeper 
tissues has no basis in published work.  Paraformaldehyde is an insoluble 
polymer, not non polymerized formaldehyde.  There is no such thing as 4% 
paraformaldehyde! 
 
It is a sad fact that many labs do not contain even one book about 
histotechnology. Nearly all books in the field (and there are many) have plenty 
of references to review articles and original literature about the techniques. 
There are also several websites that provide links to useful papers.  Check out 
some of the useful links on the Biological Stain Commission's site:  
http://biologicalstaincommission.org/useful-links/  
 
As a graduate student, you  need to  work from primary sources or reliable 
secondary sources. When you defend your thesis, you won't want to justify your 
fixation or staining method by saying I got the method by asking on an 
internet listserver. 
 
John Kiernan
Professor Emeritus
Anatomy  Cell Biology
University of Western Ontario
London,Canada
= = = 

On 05/06/15, Peter Noyce pwno...@gmail.com wrote: 
  Formaldehyde 37% (commonly called 100% formalin) compared to 4% ( commonly
 known as 10% neutral buffered formalin)-in theory the 37% should fix quicker
 and better BUT anecdotally it is said that high concentrations of
 formaldehyde quickly form a shell in the tissue and will stop good
 penetration and fixation to the deeper tissues AND over the years it has
 been said anecdotally that 4%  concentration is the quickest and most
 complete for all sample (mammal and plant) fixation and preservation-are
 these true. Please do discuss the methanol or buffers that is in the
 formaldehyde, or discuss paraformaldehyde (which is non polymerized
 formaldehyde with no methanol, in water).
 
 Regards Peter Noyce PhD student.
 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 
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Re: [Histonet] tissue fixation-formaldehyde concentrations which is best.

[Tony Henwood (SCHN)]

John,

I totally agree

Tony

From: John Kiernan [jkier...@uwo.ca]
Sent: Saturday, 6 June 2015 2:31 PM
To: Peter Noyce; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] tissue fixation-formaldehyde concentrations which   
is  best.

 Dear Peter,

Some of the information you mention as anecdotal is wrong. Formaldehyde and 
paraformaldehyde are well documented in original peer-reviewed papers and in 
all textbooks in the fields of histotechnology and histochemistry.

Your anecdote about high concentrations of formaldehyde quickly form a 'shell' 
in the tissue and will stop good penetration and fixation to the deeper 
tissues has no basis in published work.  Paraformaldehyde is an insoluble 
polymer, not non polymerized formaldehyde.  There is no such thing as 4% 
paraformaldehyde!

It is a sad fact that many labs do not contain even one book about 
histotechnology. Nearly all books in the field (and there are many) have plenty 
of references to review articles and original literature about the techniques. 
There are also several websites that provide links to useful papers.  Check out 
some of the useful links on the Biological Stain Commission's site:  
http://biologicalstaincommission.org/useful-links/

As a graduate student, you  need to  work from primary sources or reliable 
secondary sources. When you defend your thesis, you won't want to justify your 
fixation or staining method by saying I got the method by asking on an 
internet listserver.

John Kiernan
Professor Emeritus
Anatomy  Cell Biology
University of Western Ontario
London,Canada
= = =

On 05/06/15, Peter Noyce pwno...@gmail.com wrote:
  Formaldehyde 37% (commonly called 100% formalin) compared to 4% ( commonly
 known as 10% neutral buffered formalin)-in theory the 37% should fix quicker
 and better BUT anecdotally it is said that high concentrations of
 formaldehyde quickly form a shell in the tissue and will stop good
 penetration and fixation to the deeper tissues AND over the years it has
 been said anecdotally that 4%  concentration is the quickest and most
 complete for all sample (mammal and plant) fixation and preservation-are
 these true. Please do discuss the methanol or buffers that is in the
 formaldehyde, or discuss paraformaldehyde (which is non polymerized
 formaldehyde with no methanol, in water).

 Regards Peter Noyce PhD student.

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


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Re: [Histonet] tissue fixation-formaldehyde concentrations

[Hobbs, Carl]

I have to agree with the student, John.
Sure, he is coming from ignorance ( not a bad situation: naivety is not a 
fault...   we all are/were there at some point;-)
Sure...I agree with you re the using of the word  Paraformaldehyde as a 
fixative
I often sigh when used.

However, differential fixation ( zonal fixation) has always been an issue.
We often see the zonal effects of this?
Particularly in lymph nodes ( parenchymatous tissues)
Obviously, when immersion fixing.
Most often ...NOT with agitation.

Respectfully,

Carl 
 
Carl Hobbs FIBMS 
Histology and Imaging Manager 
Wolfson CARD 
Guys Campus, London Bridge  
Kings College London 
London 
SE1 1UL 
  
020 7848 6813
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Re: [Histonet] tissue fixation-formaldehyde concentrations which is best.

[John Kiernan]

 Dear Peter, 
 
Some of the information you mention as anecdotal is wrong. Formaldehyde and 
paraformaldehyde are well documented in original peer-reviewed papers and in 
all textbooks in the fields of histotechnology and histochemistry.  
 
Your anecdote about high concentrations of formaldehyde quickly form a 'shell' 
in the tissue and will stop good penetration and fixation to the deeper 
tissues has no basis in published work.  Paraformaldehyde is an insoluble 
polymer, not non polymerized formaldehyde.  There is no such thing as 4% 
paraformaldehyde! 
 
It is a sad fact that many labs do not contain even one book about 
histotechnology. Nearly all books in the field (and there are many) have plenty 
of references to review articles and original literature about the techniques. 
There are also several websites that provide links to useful papers.  Check out 
some of the useful links on the Biological Stain Commission's site:  
http://biologicalstaincommission.org/useful-links/  
 
As a graduate student, you  need to  work from primary sources or reliable 
secondary sources. When you defend your thesis, you won't want to justify your 
fixation or staining method by saying I got the method by asking on an 
internet listserver. 
 
John Kiernan
Professor Emeritus
Anatomy  Cell Biology
University of Western Ontario
London,Canada
= = = 

On 05/06/15, Peter Noyce pwno...@gmail.com wrote: 
  Formaldehyde 37% (commonly called 100% formalin) compared to 4% ( commonly
 known as 10% neutral buffered formalin)-in theory the 37% should fix quicker
 and better BUT anecdotally it is said that high concentrations of
 formaldehyde quickly form a shell in the tissue and will stop good
 penetration and fixation to the deeper tissues AND over the years it has
 been said anecdotally that 4%  concentration is the quickest and most
 complete for all sample (mammal and plant) fixation and preservation-are
 these true. Please do discuss the methanol or buffers that is in the
 formaldehyde, or discuss paraformaldehyde (which is non polymerized
 formaldehyde with no methanol, in water).
 
 Regards Peter Noyce PhD student.
 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 
___
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[Histonet] tissue fixation-formaldehyde concentrations which is best.

[Peter Noyce]

Formaldehyde 37% (commonly called 100% formalin) compared to 4% ( commonly
known as 10% neutral buffered formalin)-in theory the 37% should fix quicker
and better BUT anecdotally it is said that high concentrations of
formaldehyde quickly form a shell in the tissue and will stop good
penetration and fixation to the deeper tissues AND over the years it has
been said anecdotally that 4%  concentration is the quickest and most
complete for all sample (mammal and plant) fixation and preservation-are
these true. Please do discuss the methanol or buffers that is in the
formaldehyde, or discuss paraformaldehyde (which is non polymerized
formaldehyde with no methanol, in water).

Regards Peter Noyce PhD student.

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