[Histonet] [EXTERNAL] Re: Tissue fixation

Elizabeth Chlipala liz at premierlab.com
Fri Mar 31 14:55:08 CDT 2017


I'm going to give my two cents here.  I think you need to look closely at how you handle these samples.  First of all leeps are not small samples therefore just because the leep has been placed in fixative upon removal and then sits in fixative until its processed.  Unless the sample is grossed  into smaller pieces only then it will adequately fix.   Electro cautery  instruments function to do two things, they cut and then cauterize - there is always a thermal defect associated with the process or thermal spread, this is clearly evident on the H&E slides.   The process is essentially coagulating the tissue proteins.  The thermal spread and char that is associated with this process varies dependent upon the instrument and setting that is being used.  This process may decrease fixative penetration so it's important that these samples are grossed in as soon as possible.  Trimming the tissue will allow for better penetration of the fixative and good fixation requires at least 4 to 6 hours in formalin AFTER the tissue has been trimmed in.    The artifact that the pathologist is seeing might not be related to the amount of time the sample sits in formalin it might be due to the amount of time the samples sits  in formalin prior to being grossed in.    

If I had this issue I would start tracking the different times associated with these samples and possibly the instrument and setting used (if you are able to get that info).   

1.  Time to fixative - cold ischemic time
2.  Time to when the sample is grossed in 
3.  Time in formalin once the sample is grossed in

Good Luck I hope this helps.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz at premierlab.com
www.premierlab.com

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-----Original Message-----
From: Stedman, Nancy via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Friday, March 31, 2017 12:40 PM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] [EXTERNAL] Re: Tissue fixation

I would bet there are quite a few pathologists on this mailing list (like me) who are here because we respect histo techs and know you have valuable information to offer!

I agree, cautery artifact is pretty easy to tell from poor fixation, so it should be easy to determine if that is the problem.  Sometimes, specimens that are submitted smashed in a cassette may have poor fixation of the tissue in contact with the cassette.  Or if they are submitted sponged and floating, some tissue may not be fully immersed in formalin.  Also, crush artifact may resemble poor fixation, so maybe the clinical team is not handling the tissue gently enough?  

I read veterinary specimens; we don't do LEEPs so I don't know if any of these scenarios are possible, but they are things I see frequently.  

-Nancy Stedman


-----Original Message-----
From: Mayer,Toysha N via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Friday, March 31, 2017 2:15 PM
To: 'histonet at lists.utsouthwestern.edu'
Subject: [EXTERNAL] Re: [Histonet] Tissue fixation

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 +0000
From: T H <thigginsht at msn.com>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Tissue Fixation
Message-ID:
	<SN1PR19MB060735D7B896C67B0223E098D8370 at 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 +0000 (UTC)
From: Rene J Buesa <rjbuesa at yahoo.com>
To: T H <thigginsht at msn.com>, 	"histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation
Message-ID: <1730848134.8065268.1490967155655 at 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 at 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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------------------------------

Message: 5
Date: Fri, 31 Mar 2017 13:57:30 +0000 (UTC)
From: Paula Keene Pierce <paula at excaliburpathology.com>
To: Rene J Buesa <rjbuesa at yahoo.com>, 	Histonet Histonet
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation
Message-ID: <372734167.8016960.1490968650877 at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

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 at lists.utsouthwestern.edu>
 To: T H <thigginsht at msn.com>; "histonet at lists.utsouthwestern.edu" <histonet at 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?Ren? 

? ? On Friday, March 31, 2017 9:11 AM, T H via Histonet <histonet at 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
  


Message: 6
Date: Fri, 31 Mar 2017 15:06:53 +0000
From: "Morken, Timothy" <Timothy.Morken at ucsf.edu>
To: Rene J Buesa <rjbuesa at yahoo.com>, T H <thigginsht at msn.com>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Tissue Fixation
Message-ID:
	<761E2B5697F795489C8710BCC72141FF8E5C6461 at ex07.net.ucsf.edu>
Content-Type: text/plain; charset="utf-8"

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



Content-Type: text/plain; charset="utf-8"

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.weems at emoryhealthcare.org



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