[Histonet] Nuclear Bubbling

Aubrey Wanner Aubrey at nsh.org
Wed Feb 17 12:00:27 CST 2016


NSH has an archived webinar: Nuclear Bubbling: It's About Time" created by Peggy Wenk.  The webinar talks about several causes of nuclear bubbling, changes that can be made to reduce this problem, and, for really bad cases, Peggy talks about how to dry the slide differently, reprocess tissue or use a HIER technique to improve the quality of the nuclear detail.  

It's available to members through our online community: The Block: theblock.nsh.org 

Aubrey M.J. Wanner
National Society for Histotechnology



-----Original Message-----
From: histonet-request at lists.utsouthwestern.edu [mailto:histonet-request at lists.utsouthwestern.edu] 
Sent: Wednesday, February 17, 2016 12:04 PM
To: histonet at lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 147, Issue 16

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Today's Topics:

   1. Sakura Pre-Processing Fixative (7117) for X120 Processors
      (Jonathan Jennings)
   2. Re: Nuclear Bubbling (Rene J Buesa)
   3. (CLRW) Clinical Laboratory Reagent Water (Brent Adams)
   4. Re: Nuclear Bubbling (Jennifer MacDonald)
   5. Re: Nuclear Bubbling (Manfre, Philip)
   6. Re: Nuclear Bubbling (Jamal Rowaihi)
   7. Re: Nuclear Bubbling (Vickroy, James)
   8. nuclear bubbling (Carlos Defeo)
   9. Re: Nuclear Bubbling (Katie Sands)
  10. Sequenza Immunostaining Racks (Carlos Genty)
  11. Error Prevention: (Jb)


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

Message: 1
Date: Tue, 16 Feb 2016 18:00:59 +0000
From: Jonathan Jennings <JJennings at thedermlab.com>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Sakura Pre-Processing Fixative (7117) for X120
	Processors
Message-ID:
	<248395418C8C074A83C7D06081C4F01634741ADB at DERMLAB-SBS.dermlab.local>
Content-Type: text/plain; charset="us-ascii"

Hello All,

I manage a histology  Dermpath lab  (We only process skins) and we use Sakura X120 Processors. We have been doing excellent with the use of our x120 processors that allowed use to have slides on the Pathologist desk within 5 hours from the time we received the specimens.  That said, there is always room for improvement!  As anyone who uses the Sakura x120 Xpress processors know, all of the solutions and reagents required are "proprietary" so I don't know exactly what is in them.  To top that off, we bought the processors from a 3rd party vendor, so Sakura isn't too quick to come out and help me with this.  All of That noted, the reason I am sending this email is because I want to incorporate the pre-processing fixative because the specimens we receive in the afternoon haven't been in formalin as long (3-5 hours) as the specimens that we receive by mail in the morning (in formalin for 18+ hours).  Our preprocessing solution is working ok, but I read in the user manual about  preprocessing fixative that should be used instead, if the specimens were not completely fixed before processing.

Here are a few details of our current processing workflow with only using preprocessing solution.

*         When grossing we have 3 small stirring hot plates set at 38 Celsius. Each stir plate described below

o   1  for the 0.2 or < Biopsies filled with 10% NBF prior to  preprocessing solution for 15 minutes then processed on standard (1 hour program)

o   1  for the 0.3 or >  Biopsies Filled with Statfix (statlab) Must be in statfix. Cassettes must be in for at least 30 minutes prior to preprocessing solution (30 minutes) Then Placed on the processor (Extended - 2 hour program)

o   1 for the excisions, Cyst and all other fatty skin specimens, filled with statfix  Cassettes must be in for at least 1 hour prior to preprocessing solution (30 minutes)  The placed on processor (Extended - 2 hour program)

I tried the preprocessing fixative (no heat, just stirring agitation) on test excisions and it definitely made the tissue more firm, (almost too hard) for 30 minutes.  We had to use preprocessing solution for 1 hour on excisions, so that is a 30 minute improvement.  It also turned the tissue yellow, which was weird.  Before I try to test more tissue, I wanted to see if anyone else has went through the same preprocessing fixative testing with skins.

Questions  Finally! ;)

*         Does anyone use the preprocessing fixative for skin specimens and would you mind giving me some insight/advice on how to incorporate?

*         Is it ok to incorporate any heat to the preprocessing solution or preprocessing fixative to speed up processing time?

*         Does the preprocessing fixative affect IHCs any differently than the preprocessing solution?

*         Does anyone have any comments/ideas about my inquiry?

I apologize for the lengthy post.  Thank you for taking the time to read it.  I hope everyone has a blessed day!




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

Message: 2
Date: Tue, 16 Feb 2016 18:12:27 +0000 (UTC)
From: Rene J Buesa <rjbuesa at yahoo.com>
To: "Vickroy, James" <jvickroy at SpringfieldClinic.com>,
	"histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
	<656599459.4038172.1455646347478.JavaMail.yahoo at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

If I remember correctly, this issue has been discussed previously.The general consensus as to the cause of nuclear "bubbling" (in reality a lack of staining in the nuclear area) has been attributed to an incomplete section drying.After the section has be "fished" from the water bath, if the slide is not set to drain the underneath water before drying, the nuclear components are dissolved hence when the section is stained, there is nothing to stain ? "nuclear bubbling".I think this has been previously stated so I really do not understand posting this same question again.I do not think that posting again the question a different answer is going to be received.ren? 

    On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <histonet at lists.utsouthwestern.edu> wrote:
 

 
Struggling to find an answer.? We do a lot of GI biopsies in our lab.? Sometimes they look wonderful without any nuclear bubbling, other times the bubbling is pretty intense.? Since nuclear bubbling is often attributed to incomplete fixation we of course have investigated the fixation times.? I do not find that the problem is fixation.? In fact some of the biopsies end up fixing for 48 hrs before processing. (weekend).? There was a suggestion last week or so that there might be water trapped under the slides after cutting and before staining.? I really thought that this might be the issue however I'm not sure at this point.? Extra drying seems to help but sometimes slides side by side are so variable, one with bubbles and one without.? I also don't believe the problem is in the processing schedule since the problem has shown up on both a rapid and a normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.? Here are my questions:


1.? ? ? ? Could it be something that is happening with the tissue before it gets to the lab?? Usually a delay if fixation? causes other artifacts but not bubbling.? Could it be heat from the GI procedure?

2.? ? ? We do use blue sponges for our biopsies.? I know some say get rid of the sponges but has anyone seen this problem caused by usage of sponges?

3.? ? ? What about the heat stage in our Prisma stainer?


I am really getting frustrated.? Pathologists never complain however I would rather all of the tissue did not have the "nuclear bubbling".? Again we only do biopsies so I really don't think the standard old " not enough time in formalin" is the issue.? I have even wondered about variables such as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois? 62703
Office:? 217-528-7541, Ext. 15121
Email:? jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>



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

Message: 3
Date: Tue, 16 Feb 2016 18:34:46 +0000
From: Brent Adams <badams at acadianagastro.com>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] (CLRW) Clinical Laboratory Reagent Water
Message-ID:
	<CY1PR0301MB164280A887A8F988B931E45BB1AD0 at CY1PR0301MB1642.namprd03.prod.outlook.com>
	
Content-Type: text/plain; charset="Windows-1252"

Deanne,

I purchase 5 liter cubes from Mercedes Medical.
They can provide test results on each batch of water and fax to you.

I did have a CLIA inspector advise to document daily on the water as is
written on the cube for clarity of water, no sediments or signs of contamination.

Think 5 Liters is about $12.50 and I use a little less than two a month.


Brent Adams ? BS, LPN, HT


www.acadianagastro.com

Acadiana Gastroenterology Associates, LLC
439 Heymann Blvd
Lafayette, LA  70503

tel:  (337) 269-1126
fax:  (337) 269-1476

________________________________________
From: histonet-request at lists.utsouthwestern.edu <histonet-request at lists.utsouthwestern.edu>
Sent: Tuesday, February 16, 2016 12:00 PM
To: histonet at lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 147, Issue 15

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        histonet at lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
        http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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When replying, please edit your Subject line so it is more specific
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Today's Topics:

   1. (CLRW) - Clinical Laboratory Reagent Water (Knutson, Deanne)
   2. Tissue cassette baskets for VIP 6 (Vickroy, James)
   3. MICROTOME KNIFE SHARPENING (Hannen, Valerie)
   4. Re: (CLRW) - Clinical Laboratory Reagent Water (Morken, Timothy)
   5. Nuclear Bubbling (Vickroy, James)
   6. Complementary webinar on post-processing scientific       images,
      Feb 24 @ 1PM EST (J. Sedgewick)


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

Message: 1
Date: Mon, 15 Feb 2016 12:14:06 -0600
From: "Knutson, Deanne" <DKnutson at primecare.org>
To: "'histonet at lists.utsouthwestern.edu'"
        <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] (CLRW) - Clinical Laboratory Reagent Water
Message-ID:
        <1E0E2B14C709174B8AC2BE0AE7F76833A4D5970FFC at EXCHANGE2K7.staprimecare.org>

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

Fellow Histonetters,

Our medical center will no longer be providing us with water for our laboratory procedures.
I was wondering where other labs who need to purchase Clinical Laboratory Reagent Water are doing so?

Thank you for your help!

Deanne Knutson
Supervisor
Anatomic Pathology




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

Message: 2
Date: Mon, 15 Feb 2016 19:12:06 +0000
From: "Vickroy, James" <jvickroy at SpringfieldClinic.com>
To: "histonet at lists.utsouthwestern.edu"
        <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Tissue cassette baskets for VIP 6
Message-ID:
        <9B1A1501A800064397369BD8072E6BCA06502BD8 at E2K10DB.springfieldclinic.com>

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


Anybody have an idea where we can get a used cassette basket that will fit in the VIP 6?  The cost of a new one is pretty high?

Jim

Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>



This electronic message contains information from Springfield Clinic, LLP that may be confidential, privileged, and/or sensitive. This information is intended for the use of the individual(s) or entity(ies) named above. If you are not the intended recipient, be aware that disclosure, copying, distribution, or action taken on the contents of this information is strictly prohibited. If you have received this electronic message in error, please notify the sender immediately, by electronic mail, so that arrangements may be made for the retrieval of this electronic message. Thank you.

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

Message: 3
Date: Tue, 16 Feb 2016 10:28:59 -0500
From: "Hannen, Valerie" <Valerie.Hannen at parrishmed.com>
To: "Histonet at lists.utsouthwestern.edu"
        <Histonet at lists.utsouthwestern.edu>
Subject: [Histonet] MICROTOME KNIFE SHARPENING
Message-ID: <450B7A81EDA0C54E97C53D60F00776C323546750BC at isexstore03>
Content-Type: text/plain; charset="us-ascii"

Hi all...  Hoping you might be answer a couple of questions that I have.  1) Does anyone use a microtome knife sharpening kit made by Pathco?

If so, 2) How well does it sharpen the blades?  3) Is there any issue on attaching the abrasive sheets to the honing plate?  4) Do the abrasive sheets tend to "move"

during the sharpening process, that would cause the blade not be on them but on the plate instead?

  I currently am using coarse and fine abrasive liquids along with the honing plate... but the coarse, fine and honing liquids are becoming quite expensive and hard to come by.

Any and all replies are welcomed.

Thank you in advance,



Valerie Hannen,MLT(ASCP),HTL,SU (FL)
Section Chief, Histology
Parrish Medical Center
951 N. Washington Ave.
Titusville,Florida 32796
T: (321)268-6333 ext. 7506
F: (321) 268-6149
valerie.hannen at parrishmed.com<mailto:valerie.hannen at parrishmed.com>
www.parrishmed.com

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Message: 4
Date: Tue, 16 Feb 2016 16:05:12 +0000
From: "Morken, Timothy" <Timothy.Morken at ucsf.edu>
To: "Knutson, Deanne" <DKnutson at primecare.org>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] (CLRW) - Clinical Laboratory Reagent Water
Message-ID:
        <761E2B5697F795489C8710BCC72141FF6FD17480 at ex07.net.ucsf.edu>
Content-Type: text/plain; charset="us-ascii"

Deanne,

If you just need it for a few critical reagents You can get type 1 water by the pint or the gallon from Fisher (NERL Type 1 water). It's probably overkill for most histology procedures, but convenient if you need it. Note that the expiry clock of 30 days starts when you open the bottle.


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



-----Original Message-----
From: Knutson, Deanne via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Monday, February 15, 2016 10:14 AM
To: 'histonet at lists.utsouthwestern.edu'
Subject: [Histonet] (CLRW) - Clinical Laboratory Reagent Water

Fellow Histonetters,

Our medical center will no longer be providing us with water for our laboratory procedures.
I was wondering where other labs who need to purchase Clinical Laboratory Reagent Water are doing so?

Thank you for your help!

Deanne Knutson
Supervisor
Anatomic Pathology




  ________________________________
This email may include confidential and privileged information. If this is not intended for your use, please destroy immediately and contact the sender of the message.

This email and attachments contain information that may be confidential or privileged. If you are not the intended recipient, notify the sender at once and delete this message completely from your information system. Further use, disclosure, or copying of information contained in this email is not authorized, and any such action should not be construed as a waiver of privilege or other confidentiality protections.
_______________________________________________
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Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 5
Date: Tue, 16 Feb 2016 17:10:40 +0000
From: "Vickroy, James" <jvickroy at SpringfieldClinic.com>
To: "histonet at lists.utsouthwestern.edu"
        <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Nuclear Bubbling
Message-ID:
        <9B1A1501A800064397369BD8072E6BCA06503B35 at E2K10DB.springfieldclinic.com>

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


Struggling to find an answer.  We do a lot of GI biopsies in our lab.   Sometimes they look wonderful without any nuclear bubbling, other times the bubbling is pretty intense.  Since nuclear bubbling is often attributed to incomplete fixation we of course have investigated the fixation times.  I do not find that the problem is fixation.  In fact some of the biopsies end up fixing for 48 hrs before processing. (weekend).  There was a suggestion last week or so that there might be water trapped under the slides after cutting and before staining.   I really thought that this might be the issue however I'm not sure at this point.  Extra drying seems to help but sometimes slides side by side are so variable, one with bubbles and one without.   I also don't believe the problem is in the processing schedule since the problem has shown up on both a rapid and a normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.  Here are my questions:


1.        Could it be something that is happening with the tissue before it gets to the lab?  Usually a delay if fixation  causes other artifacts but not bubbling.  Could it be heat from the GI procedure?

2.       We do use blue sponges for our biopsies.  I know some say get rid of the sponges but has anyone seen this problem caused by usage of sponges?

3.       What about the heat stage in our Prisma stainer?


I am really getting frustrated.  Pathologists never complain however I would rather all of the tissue did not have the "nuclear bubbling".   Again we only do biopsies so I really don't think the standard old " not enough time in formalin" is the issue.  I have even wondered about variables such as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>



This electronic message contains information from Springfield Clinic, LLP that may be confidential, privileged, and/or sensitive. This information is intended for the use of the individual(s) or entity(ies) named above. If you are not the intended recipient, be aware that disclosure, copying, distribution, or action taken on the contents of this information is strictly prohibited. If you have received this electronic message in error, please notify the sender immediately, by electronic mail, so that arrangements may be made for the retrieval of this electronic message. Thank you.

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

Message: 6
Date: Tue, 16 Feb 2016 11:31:27 -0600
From: "J. Sedgewick" <jerrysedgewick at gmail.com>
To: <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Complementary webinar on post-processing
        scientific      images, Feb 24 @ 1PM EST
Message-ID: <1557B579CB2A478B884FD20B9E5064D6 at sedge>
Content-Type: text/plain; format=flowed; charset="UTF-8";
        reply-type=original

Hello All,

Do you post-process your scientific images?  Typical tools for common
adjustments include Photoshop and ImageJ, but they aren?t the best answer.
You are invited to attend a complementary webinar on Feb 24 at 1:00PM EST ?
?Best Practices for Post-Processing of Scientific Images?

Reserve your webinar seat now at
https://attendee.gotowebinar.com/register/1379379490730827010

I'll be giving this webinar. Also, a similar presentation will be given for
the histology WOW 2016 with Dr. Michael Linden at the University of
Minnesota on March 25.

Best,
Jerry Sedgewick






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

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

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Message: 4
Date: Tue, 16 Feb 2016 11:33:47 -0800
From: Jennifer MacDonald <JMacDonald at mtsac.edu>
To: Rene J Buesa <rjbuesa at yahoo.com>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>,	"Vickroy, James"
	<jvickroy at SpringfieldClinic.com>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
	<OF6CCD2B49.A88FCD99-ON88257F5B.006B4E2B-88257F5B.006B76AD at mtsac.edu>
Content-Type: text/plain; charset="GB2312"

There is no need to be rude.  He has tried the drying option and is still 
having nuclear bubbling.  He is exploring other possible issues.  You 
would see this if you read the email in its entirety. 



From:   Rene J Buesa via Histonet <histonet at lists.utsouthwestern.edu>
To:     "Vickroy, James" <jvickroy at SpringfieldClinic.com>, 
"histonet at lists.utsouthwestern.edu" <histonet at lists.utsouthwestern.edu>
Date:   02/16/2016 10:13 AM
Subject:        Re: [Histonet] Nuclear Bubbling



If I remember correctly, this issue has been discussed previously.The 
general consensus as to the cause of nuclear "bubbling" (in reality a lack 
of staining in the nuclear area) has been attributed to an incomplete 
section drying.After the section has be "fished" from the water bath, if 
the slide is not set to drain the underneath water before drying, the 
nuclear components are dissolved hence when the section is stained, there 
is nothing to stain ? "nuclear bubbling".I think this has been previously 
stated so I really do not understand posting this same question again.I do 
not think that posting again the question a different answer is going to 
be received.ren? 

    On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
<histonet at lists.utsouthwestern.edu> wrote:
 

 
Struggling to find an answer.  We do a lot of GI biopsies in our lab.  
Sometimes they look wonderful without any nuclear bubbling, other times 
the bubbling is pretty intense.  Since nuclear bubbling is often 
attributed to incomplete fixation we of course have investigated the 
fixation times.  I do not find that the problem is fixation.  In fact some 
of the biopsies end up fixing for 48 hrs before processing. (weekend).  
There was a suggestion last week or so that there might be water trapped 
under the slides after cutting and before staining.  I really thought that 
this might be the issue however I'm not sure at this point.  Extra drying 
seems to help but sometimes slides side by side are so variable, one with 
bubbles and one without.  I also don't believe the problem is in the 
processing schedule since the problem has shown up on both a rapid and a 
normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.  Here are my 
questions:


1.        Could it be something that is happening with the tissue before 
it gets to the lab?  Usually a delay if fixation  causes other artifacts 
but not bubbling.  Could it be heat from the GI procedure?

2.      We do use blue sponges for our biopsies.  I know some say get rid 
of the sponges but has anyone seen this problem caused by usage of 
sponges?

3.      What about the heat stage in our Prisma stainer?


I am really getting frustrated.  Pathologists never complain however I 
would rather all of the tissue did not have the "nuclear bubbling".  Again 
we only do biopsies so I really don't think the standard old " not enough 
time in formalin" is the issue.  I have even wondered about variables such 
as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<
mailto:jvickroy at SpringfieldClinic.com>



This electronic message contains information from Springfield Clinic, LLP 
that may be confidential, privileged, and/or sensitive. This information 
is intended for the use of the individual(s) or entity(ies) named above. 
If you are not the intended recipient, be aware that disclosure, copying, 
distribution, or action taken on the contents of this information is 
strictly prohibited. If you have received this electronic message in 
error, please notify the sender immediately, by electronic mail, so that 
arrangements may be made for the retrieval of this electronic message. 
Thank you.
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


 
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 5
Date: Tue, 16 Feb 2016 14:44:52 -0500
From: "Manfre, Philip" <philip_manfre at merck.com>
To: Rene J Buesa <rjbuesa at yahoo.com>, "Vickroy, James"
	<jvickroy at SpringfieldClinic.com>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
	<558A4571351D0C42BD923F403F4198C40109FE02D4DF at USCTMXP51014.merck.com>
Content-Type: text/plain; charset="utf-8"

Sort of a rude response to someone looking for help.

-----Original Message-----
From: Rene J Buesa via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Tuesday, February 16, 2016 1:12 PM
To: Vickroy, James; histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Nuclear Bubbling

If I remember correctly, this issue has been discussed previously.The general consensus as to the cause of nuclear "bubbling" (in reality a lack of staining in the nuclear area) has been attributed to an incomplete section drying.After the section has be "fished" from the water bath, if the slide is not set to drain the underneath water before drying, the nuclear components are dissolved hence when the section is stained, there is nothing to stain ? "nuclear bubbling".I think this has been previously stated so I really do not understand posting this same question again.I do not think that posting again the question a different answer is going to be received.ren? 

    On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <histonet at lists.utsouthwestern.edu> wrote:
 

 
Struggling to find an answer.? We do a lot of GI biopsies in our lab.? Sometimes they look wonderful without any nuclear bubbling, other times the bubbling is pretty intense.? Since nuclear bubbling is often attributed to incomplete fixation we of course have investigated the fixation times.? I do not find that the problem is fixation.? In fact some of the biopsies end up fixing for 48 hrs before processing. (weekend).? There was a suggestion last week or so that there might be water trapped under the slides after cutting and before staining.? I really thought that this might be the issue however I'm not sure at this point.? Extra drying seems to help but sometimes slides side by side are so variable, one with bubbles and one without.? I also don't believe the problem is in the processing schedule since the problem has shown up on both a rapid and a normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.? Here are my questions:


1.? ? ? ? Could it be something that is happening with the tissue before it gets to the lab?? Usually a delay if fixation? causes other artifacts but not bubbling.? Could it be heat from the GI procedure?

2.? ? ? We do use blue sponges for our biopsies.? I know some say get rid of the sponges but has anyone seen this problem caused by usage of sponges?

3.? ? ? What about the heat stage in our Prisma stainer?


I am really getting frustrated.? Pathologists never complain however I would rather all of the tissue did not have the "nuclear bubbling".? Again we only do biopsies so I really don't think the standard old " not enough time in formalin" is the issue.? I have even wondered about variables such as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois? 62703
Office:? 217-528-7541, Ext. 15121
Email:? jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>



This electronic message contains information from Springfield Clinic, LLP that may be confidential, privileged, and/or sensitive. This information is intended for the use of the individual(s) or entity(ies) named above. If you are not the intended recipient, be aware that disclosure, copying, distribution, or action taken on the contents of this information is strictly prohibited. If you have received this electronic message in error, please notify the sender immediately, by electronic mail, so that arrangements may be made for the retrieval of this electronic message. Thank you.
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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

Message: 6
Date: Tue, 16 Feb 2016 22:56:28 +0300
From: Jamal Rowaihi <j.rowaihi at alborglaboratories.com>
To: "Manfre, Philip" <philip_manfre at merck.com>, Rene J Buesa
	<rjbuesa at yahoo.com>, "Vickroy, James" <jvickroy at SpringfieldClinic.com>
Cc: ???? ??????? 	<j.rowaihi at alborglaboratories.com>,
	"histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID: <pwsp90twv5eo949blvukh4jo.1455652588197 at email.android.com>
Content-Type: text/plain; charset=utf-8

Great, I agree?


Regards
Jamal RowaihiAnatomic Pathology SupervisorAl Borg Medical Laboratories?Sent from my cell phone-------- Original message --------From: "Manfre, Philip via Histonet" <histonet at lists.utsouthwestern.edu> Date: 2/16/2016  10:44 PM  (GMT+03:00) To: Rene J Buesa <rjbuesa at yahoo.com>, "Vickroy, James" <jvickroy at SpringfieldClinic.com> Cc: histonet at lists.utsouthwestern.edu Subject: Re: [Histonet] Nuclear Bubbling 
Sort of a rude response to someone looking for help.

-----Original Message-----
From: Rene J Buesa via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Tuesday, February 16, 2016 1:12 PM
To: Vickroy, James; histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Nuclear Bubbling

If I remember correctly, this issue has been discussed previously.The general consensus as to the cause of nuclear "bubbling" (in reality a lack of staining in the nuclear area) has been attributed to an incomplete section drying.After the section has be "fished" from the water bath, if the slide is not set to drain the underneath water before drying, the nuclear components are dissolved hence when the section is stained, there is nothing to stain ? "nuclear bubbling".I think this has been previously stated so I really do not understand posting this same question again.I do not think that posting again the question a different answer is going to be received.ren? 

??? On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <histonet at lists.utsouthwestern.edu> wrote:
 

 
Struggling to find an answer.? We do a lot of GI biopsies in our lab.? Sometimes they look wonderful without any nuclear bubbling, other times the bubbling is pretty intense.? Since nuclear bubbling is often attributed to incomplete fixation we of course have investigated the fixation times.? I do not find that the problem is fixation.? In fact some of the biopsies end up fixing for 48 hrs before processing. (weekend).? There was a suggestion last week or so that there might be water trapped under the slides after cutting and before staining.? I really thought that this might be the issue however I'm not sure at this point.? Extra drying seems to help but sometimes slides side by side are so variable, one with bubbles and one without.? I also don't believe the problem is in the processing schedule since the problem has shown up on both a rapid and a normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.? Here are my questions:


1.? ? ? ? Could it be something that is happening with the tissue before it gets to the lab?? Usually a delay if fixation? causes other artifacts but not bubbling.? Could it be heat from the GI procedure?

2.? ? ? We do use blue sponges for our biopsies.? I know some say get rid of the sponges but has anyone seen this problem caused by usage of sponges?

3.? ? ? What about the heat stage in our Prisma stainer?


I am really getting frustrated.? Pathologists never complain however I would rather all of the tissue did not have the "nuclear bubbling".? Again we only do biopsies so I really don't think the standard old " not enough time in formalin" is the issue.? I have even wondered about variables such as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois? 62703
Office:? 217-528-7541, Ext. 15121
Email:? jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>



This electronic message contains information from Springfield Clinic, LLP that may be confidential, privileged, and/or sensitive. This information is intended for the use of the individual(s) or entity(ies) named above. If you are not the intended recipient, be aware that disclosure, copying, distribution, or action taken on the contents of this information is strictly prohibited. If you have received this electronic message in error, please notify the sender immediately, by electronic mail, so that arrangements may be made for the retrieval of this electronic message. Thank you.
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


? 
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Notice:? This e-mail message, together with any attachments, contains
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------------------------------

Message: 7
Date: Tue, 16 Feb 2016 21:53:00 +0000
From: "Vickroy, James" <jvickroy at SpringfieldClinic.com>
To: 'Jamal Rowaihi' <j.rowaihi at alborglaboratories.com>, "Manfre,
	Philip"	<philip_manfre at merck.com>, Rene J Buesa <rjbuesa at yahoo.com>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
	<9B1A1501A800064397369BD8072E6BCA06504834 at E2K10DB.springfieldclinic.com>
	
Content-Type: text/plain; charset="utf-8"

For the record please note that I have over thirty-six years experience working in a Histology lab.   I have been a supervisor or manager  of a hospital and clinic histology department for at least 25 years.

Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>


From: Jamal Rowaihi [mailto:j.rowaihi at alborglaboratories.com]
Sent: Tuesday, February 16, 2016 1:56 PM
To: Manfre, Philip; Rene J Buesa; Vickroy, James
Cc: ???? ???????; histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Nuclear Bubbling

Great, I agree



Regards

Jamal Rowaihi
Anatomic Pathology Supervisor
Al Borg Medical Laboratories
Sent from my cell phone
-------- Original message --------
From: "Manfre, Philip via Histonet" <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>>
Date: 2/16/2016 10:44 PM (GMT+03:00)
To: Rene J Buesa <rjbuesa at yahoo.com<mailto:rjbuesa at yahoo.com>>, "Vickroy, James" <jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>>
Cc: histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling

Sort of a rude response to someone looking for help.

-----Original Message-----
From: Rene J Buesa via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, February 16, 2016 1:12 PM
To: Vickroy, James; histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling

If I remember correctly, this issue has been discussed previously.The general consensus as to the cause of nuclear "bubbling" (in reality a lack of staining in the nuclear area) has been attributed to an incomplete section drying.After the section has be "fished" from the water bath, if the slide is not set to drain the underneath water before drying, the nuclear components are dissolved hence when the section is stained, there is nothing to stain ? "nuclear bubbling".I think this has been previously stated so I really do not understand posting this same question again.I do not think that posting again the question a different answer is going to be received.ren?

    On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>> wrote:



Struggling to find an answer.  We do a lot of GI biopsies in our lab.  Sometimes they look wonderful without any nuclear bubbling, other times the bubbling is pretty intense.  Since nuclear bubbling is often attributed to incomplete fixation we of course have investigated the fixation times.  I do not find that the problem is fixation.  In fact some of the biopsies end up fixing for 48 hrs before processing. (weekend).  There was a suggestion last week or so that there might be water trapped under the slides after cutting and before staining.  I really thought that this might be the issue however I'm not sure at this point.  Extra drying seems to help but sometimes slides side by side are so variable, one with bubbles and one without.  I also don't believe the problem is in the processing schedule since the problem has shown up on both a rapid and a normal schedule. (therefore longer dehydration, clearing, etc.)

I am wondering if anyone else has worked with this issue.  Here are my questions:


1.        Could it be something that is happening with the tissue before it gets to the lab?  Usually a delay if fixation  causes other artifacts but not bubbling.  Could it be heat from the GI procedure?

2.      We do use blue sponges for our biopsies.  I know some say get rid of the sponges but has anyone seen this problem caused by usage of sponges?

3.      What about the heat stage in our Prisma stainer?


I am really getting frustrated.  Pathologists never complain however I would rather all of the tissue did not have the "nuclear bubbling".  Again we only do biopsies so I really don't think the standard old " not enough time in formalin" is the issue.  I have even wondered about variables such as we use recycled formalin, recycled Clearite III.

Any suggestions?

Jim



Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com%3cmailto:jvickroy at SpringfieldClinic.com>>



This electronic message contains information from Springfield Clinic, LLP that may be confidential, privileged, and/or sensitive. This information is intended for the use of the individual(s) or entity(ies) named above. If you are not the intended recipient, be aware that disclosure, copying, distribution, or action taken on the contents of this information is strictly prohibited. If you have received this electronic message in error, please notify the sender immediately, by electronic mail, so that arrangements may be made for the retrieval of this electronic message. Thank you.
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu<mailto:Histonet at lists.utsouthwestern.edu>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



_______________________________________________
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Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (2000 Galloping Hill Road, Kenilworth,
New Jersey, USA 07033), and/or its affiliates Direct contact information
for affiliates is available at
http://www.merck.com/contact/contacts.html) that may be confidential,
proprietary copyrighted and/or legally privileged. It is intended solely
for the use of the individual or entity named on this message. If you are
not the intended recipient, and have received this message in error,
please notify us immediately by reply e-mail and then delete it from
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------------------------------

Message: 8
Date: Tue, 16 Feb 2016 21:56:49 +0000
From: "Carlos Defeo" <latecor at adinet.com.uy>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Cc: histonet at lists.utsouthwestern.edu
Subject: [Histonet] nuclear bubbling
Message-ID: <em1942d68c-0573-4d27-a81c-0bed93cc738c at calipc>
Content-Type: text/plain; format=flowed; charset=utf-8

Dear James:
Two chained events take place on your issue:
1-sections not entirely drained,water remains even minimally under 
sections;
2- you put these sections in the oven, the heat literally "explodes" the 
nuclear bubbles and creates a hole with no cromatin to stain.

My kind regards,
Carlos Defeo
Histotechnologist


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

Message: 9
Date: Tue, 16 Feb 2016 16:18:03 -0600
From: Katie Sands <derm.katiesands at gmail.com>
To: "Vickroy, James" <jvickroy at springfieldclinic.com>
Cc: Jamal Rowaihi <j.rowaihi at alborglaboratories.com>, 	"Manfre,
	Philip" <philip_manfre at merck.com>,	Rene J Buesa <rjbuesa at yahoo.com>,
	"histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
	<CAORF-nHjeSZx1sC1BviWza84O0fqJY5VJgtZ33FHFVghzEijmA at mail.gmail.com>
Content-Type: text/plain; charset=UTF-8

I don't have your answer Jim, yet I can vouch for your experience as a
supervisor since you were my boss for about two years.  Hopefully you can
get it resolved because it can be very frustrating as the tech.

On Tuesday, February 16, 2016, Vickroy, James via Histonet <
histonet at lists.utsouthwestern.edu> wrote:

> For the record please note that I have over thirty-six years experience
> working in a Histology lab.   I have been a supervisor or manager  of a
> hospital and clinic histology department for at least 25 years.
>
> Jim Vickroy
> Histology Manager
> Springfield Clinic, Main Campus, East Building
> 1025 South 6th Street
> Springfield, Illinois  62703
> Office:  217-528-7541, Ext. 15121
> Email:  jvickroy at SpringfieldClinic.com<mailto:
> jvickroy at SpringfieldClinic.com <javascript:;>>
>
>
> From: Jamal Rowaihi [mailto:j.rowaihi at alborglaboratories.com
> <javascript:;>]
> Sent: Tuesday, February 16, 2016 1:56 PM
> To: Manfre, Philip; Rene J Buesa; Vickroy, James
> Cc: ???? ???????; histonet at lists.utsouthwestern.edu <javascript:;>
> Subject: Re: [Histonet] Nuclear Bubbling
>
> Great, I agree
>
>
>
> Regards
>
> Jamal Rowaihi
> Anatomic Pathology Supervisor
> Al Borg Medical Laboratories
> Sent from my cell phone
> -------- Original message --------
> From: "Manfre, Philip via Histonet" <histonet at lists.utsouthwestern.edu
> <javascript:;><mailto:histonet at lists.utsouthwestern.edu <javascript:;>>>
> Date: 2/16/2016 10:44 PM (GMT+03:00)
> To: Rene J Buesa <rjbuesa at yahoo.com <javascript:;><mailto:
> rjbuesa at yahoo.com <javascript:;>>>, "Vickroy, James"
> <jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com
> <javascript:;>>>
> Cc: histonet at lists.utsouthwestern.edu <javascript:;><mailto:
> histonet at lists.utsouthwestern.edu <javascript:;>>
> Subject: Re: [Histonet] Nuclear Bubbling
>
> Sort of a rude response to someone looking for help.
>
> -----Original Message-----
> From: Rene J Buesa via Histonet [mailto:histonet at lists.utsouthwestern.edu
> <javascript:;>]
> Sent: Tuesday, February 16, 2016 1:12 PM
> To: Vickroy, James; histonet at lists.utsouthwestern.edu <javascript:;>
> <mailto:histonet at lists.utsouthwestern.edu <javascript:;>>
> Subject: Re: [Histonet] Nuclear Bubbling
>
> If I remember correctly, this issue has been discussed previously.The
> general consensus as to the cause of nuclear "bubbling" (in reality a lack
> of staining in the nuclear area) has been attributed to an incomplete
> section drying.After the section has be "fished" from the water bath, if
> the slide is not set to drain the underneath water before drying, the
> nuclear components are dissolved hence when the section is stained, there
> is nothing to stain ? "nuclear bubbling".I think this has been previously
> stated so I really do not understand posting this same question again.I do
> not think that posting again the question a different answer is going to be
> received.ren?
>
>     On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <
> histonet at lists.utsouthwestern.edu <javascript:;><mailto:
> histonet at lists.utsouthwestern.edu <javascript:;>>> wrote:
>
>
>
> Struggling to find an answer.  We do a lot of GI biopsies in our lab.
> Sometimes they look wonderful without any nuclear bubbling, other times the
> bubbling is pretty intense.  Since nuclear bubbling is often attributed to
> incomplete fixation we of course have investigated the fixation times.  I
> do not find that the problem is fixation.  In fact some of the biopsies end
> up fixing for 48 hrs before processing. (weekend).  There was a suggestion
> last week or so that there might be water trapped under the slides after
> cutting and before staining.  I really thought that this might be the issue
> however I'm not sure at this point.  Extra drying seems to help but
> sometimes slides side by side are so variable, one with bubbles and one
> without.  I also don't believe the problem is in the processing schedule
> since the problem has shown up on both a rapid and a normal schedule.
> (therefore longer dehydration, clearing, etc.)
>
> I am wondering if anyone else has worked with this issue.  Here are my
> questions:
>
>
> 1.        Could it be something that is happening with the tissue before
> it gets to the lab?  Usually a delay if fixation  causes other artifacts
> but not bubbling.  Could it be heat from the GI procedure?
>
> 2.      We do use blue sponges for our biopsies.  I know some say get rid
> of the sponges but has anyone seen this problem caused by usage of sponges?
>
> 3.      What about the heat stage in our Prisma stainer?
>
>
> I am really getting frustrated.  Pathologists never complain however I
> would rather all of the tissue did not have the "nuclear bubbling".  Again
> we only do biopsies so I really don't think the standard old " not enough
> time in formalin" is the issue.  I have even wondered about variables such
> as we use recycled formalin, recycled Clearite III.
>
> Any suggestions?
>
> Jim
>
>
>
> Jim Vickroy
> Histology Manager
> Springfield Clinic, Main Campus, East Building
> 1025 South 6th Street
> Springfield, Illinois  62703
> Office:  217-528-7541, Ext. 15121
> Email:  jvickroy at SpringfieldClinic.com<mailto:
> jvickroy at SpringfieldClinic.com <javascript:;><mailto:
> jvickroy at SpringfieldClinic.com <javascript:;>
> %3cmailto:jvickroy at SpringfieldClinic.com>>
>
>
>
> This electronic message contains information from Springfield Clinic, LLP
> that may be confidential, privileged, and/or sensitive. This information is
> intended for the use of the individual(s) or entity(ies) named above. If
> you are not the intended recipient, be aware that disclosure, copying,
> distribution, or action taken on the contents of this information is
> strictly prohibited. If you have received this electronic message in error,
> please notify the sender immediately, by electronic mail, so that
> arrangements may be made for the retrieval of this electronic message.
> Thank you.
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu <javascript:;><mailto:
> Histonet at lists.utsouthwestern.edu <javascript:;>>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu <javascript:;><mailto:
> Histonet at lists.utsouthwestern.edu <javascript:;>>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> Notice:  This e-mail message, together with any attachments, contains
> information of Merck & Co., Inc. (2000 Galloping Hill Road, Kenilworth,
> New Jersey, USA 07033), and/or its affiliates Direct contact information
> for affiliates is available at
> http://www.merck.com/contact/contacts.html) that may be confidential,
> proprietary copyrighted and/or legally privileged. It is intended solely
> for the use of the individual or entity named on this message. If you are
> not the intended recipient, and have received this message in error,
> please notify us immediately by reply e-mail and then delete it from
> your system.
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu <javascript:;><mailto:
> Histonet at lists.utsouthwestern.edu <javascript:;>>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu <javascript:;>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


-- 
Katie L. Sands
Histology Technician/Lab Manager
Advanced Dermatology of Southeast Missouri, PC
2116 Megan Drive Suite 102
Cape Girardeau, MO  63701
derm.katiesands at gmail.com
Work:  (573) 335-7546
Cell:  (309) 840-3799
www.dermsemo.com


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

Message: 10
Date: Wed, 17 Feb 2016 16:31:51 +0000
From: Carlos Genty <cgenty at criticalxsolutions.com>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Sequenza Immunostaining Racks
Message-ID:
	<BN3PR0101MB1089D98081A162C8CAD05163AEAE0 at BN3PR0101MB1089.prod.exchangelabs.com>
	
Content-Type: text/plain; charset="iso-8859-1"


Good Day Everyone,


I am looking to obtain used Thermo Sequenza Immunostaining Racks (about 10 or so).  Does anyone have any that they would be willing to part with?


Please feel free to contact me directly.


Thanks in advance!

Carlos

Carlos Genty
Critical X Solutions, LLC
cgenty at criticalxsolutions.com
(310) 357-1993



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

Message: 11
Date: Wed, 17 Feb 2016 10:03:45 -0700
From: Jb <craigak12 at gmail.com>
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Error Prevention:
Message-ID: <9ED89D91-CB8E-4014-B24A-6C91314414B8 at gmail.com>
Content-Type: text/plain;	charset=us-ascii

Does anyone have a policy that they are willing to share regarding tech errors, error prevention, etc?  

How do you document, retrain, write up, and terminate employees for errors? What is considered severe problems for termination, etc. 

I know it is up to the manager, I want to know what other lab standards are to help create a procedure. 

Thank you,

Sent from my iPhone


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

Subject: Digest Footer

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

End of Histonet Digest, Vol 147, Issue 16
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