Re: [Histonet] Nuclear Bubbling

[Rene J Buesa via Histonet]

Willem:
Essentially incomplete deparaffination shows a different pattern and is not 
limited to nuclei only.Again, this is what the consensus is:1- there is ALWAYS 
water left underneath the section2- that water HAS to be eliminated before the 
section is dried in the oven3- the best sectioning practices call for placing 
the slides (with the just fished sections) in a vertical position resting on 
their shortest end (1 inch) for a few minutes after the section is fished from 
the water bath. At my lab the techs were required to do that and their used to 
rest the sections around the water bath.4- AFTER the sections have been 
completely drained then they were either to the oven (60ºC for at least 7 min. 
or to the autostainer, we used a Sakura autostainer) and the sections were 
stained.
Which is the consensus as to what happens: when there is water underneath the 
whole section and it goes into the oven without properly drainage the HOT water 
will dissolve the nuclear contents something that it will NOT happen with the 
other tissue components (such as the connective tissue or nerves) because their 
chemical components are insoluble). The nuclei contain soluble chemical 
components and, as such, they will be dissolved in the hot water and if they 
are dissolved, they cannot be stained and will determine an image similar to an 
empty bubble, hence the "nuclear bubbling" artifact.
I have proven this mechanism by staining sections with different degrees of 
drainage (measured in minutes after the sections were fished) and the results 
were consistent with the explanation I offered above. The "ideal" drainage time 
is 7 minutes. Just try it yourself to dispel any of your doubts.
René 

On Thursday, February 18, 2016 3:00 AM, "Hoekert, Willem via Histonet" 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Could it be due to incomplete deparaffinization? 

Willem



Van: Vickroy, James via Histonet [histonet@lists.utsouthwestern.edu]
Verzonden: dinsdag 16 februari 2016 18:10
Aan: histonet@lists.utsouthwestern.edu
Onderwerp: [Histonet] Nuclear Bubbling

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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>



This electronic message contains information from Springfield Clinic, LLP that 
may be confidential, privileged, and/or sensitive. This information is intended 
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Re: [Histonet] Nuclear Bubbling

[Kienitz, Kari via Histonet]

incomplete deparafinization has my voteincrease times in xylene and I bet 
it goes away.  This happened to me when the paraffin that I had bought for 
years was slightly changed without notifying anyone.  If I remember right the 
polymer was increase resulting in xylene being less effective.


Kari Kienitz HT, (ASCP)
Histology Laboratory
Gastroenterology-EAST
The Oregon Clinic
 NE 99th Ave
Portland, OR  97220
503.935.8311
kkien...@orclinic.com




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From: Hoekert, Willem via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Wednesday, February 17, 2016 11:54 PM
To: Vickroy, James; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Nuclear Bubbling

Could it be due to incomplete deparaffinization?

Willem



Van: Vickroy, James via Histonet [histonet@lists.utsouthwestern.edu]
Verzonden: dinsdag 16 februari 2016 18:10
Aan: histonet@lists.utsouthwestern.edu
Onderwerp: [Histonet] Nuclear Bubbling

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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>



This electronic message contains information from Springfield Clinic, LLP that 
may be confidential, privileged, and/or sensitive. This information is intended 
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Re: [Histonet] Nuclear Bubbling

[Hoekert, Willem via Histonet]

Could it be due to incomplete deparaffinization? 

Willem



Van: Vickroy, James via Histonet [histonet@lists.utsouthwestern.edu]
Verzonden: dinsdag 16 februari 2016 18:10
Aan: histonet@lists.utsouthwestern.edu
Onderwerp: [Histonet] Nuclear Bubbling

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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>



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Midden
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Re: [Histonet] Nuclear Bubbling

[Aubrey Wanner via Histonet]

   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 + (UTC)
From: Rene J Buesa <rjbu...@yahoo.com>
To: "Vickroy, James" <jvick...@springfieldclinic.com>,
"histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Nuclear Bubbling
Message-ID:
<656599459.4038172.1455646347478.javamail.ya...@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@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:? jvick...@springfieldclinic.com<mailto:jvick...@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 e

Re: [Histonet] Nuclear Bubbling

[deGuzman, Jose R via Histonet]

Jim,

Are you drying the slides in an oven before the heat stage on the Prisma? If 
you use an oven, what is the temperature setting and for how long are slides 
kept in? Carson and Hladik recommends 60 degrees for less than 1 hour.

You mentioned, recycled formalin, what is the pH of the formalin?

I apologize for all the questions and no answers.

Jose


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Re: [Histonet] Nuclear Bubbling

[Katie Sands via Histonet]

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@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:  jvick...@springfieldclinic.com jvick...@springfieldclinic.com <javascript:;>>
>
>
> From: Jamal Rowaihi [mailto:j.rowa...@alborglaboratories.com
> <javascript:;>]
> Sent: Tuesday, February 16, 2016 1:56 PM
> To: Manfre, Philip; Rene J Buesa; Vickroy, James
> Cc: جمال الرويحي; histonet@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@lists.utsouthwestern.edu
> <javascript:;><mailto:histonet@lists.utsouthwestern.edu <javascript:;>>>
> Date: 2/16/2016 10:44 PM (GMT+03:00)
> To: Rene J Buesa <rjbu...@yahoo.com <javascript:;> rjbu...@yahoo.com <javascript:;>>>, "Vickroy, James"
> <jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com
> <javascript:;>>>
> Cc: histonet@lists.utsouthwestern.edu <javascript:;> histonet@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@lists.utsouthwestern.edu
> <javascript:;>]
> Sent: Tuesday, February 16, 2016 1:12 PM
> To: Vickroy, James; histonet@lists.utsouthwestern.edu <javascript:;>
> <mailto:histonet@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.rené
>
> On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" <
> histonet@lists.utsouthwestern.edu <javascript:;> histonet@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 a

Re: [Histonet] Nuclear Bubbling

[Vickroy, James via Histonet]

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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>


From: Jamal Rowaihi [mailto:j.rowa...@alborglaboratories.com]
Sent: Tuesday, February 16, 2016 1:56 PM
To: Manfre, Philip; Rene J Buesa; Vickroy, James
Cc: جمال الرويحي; histonet@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@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>>
Date: 2/16/2016 10:44 PM (GMT+03:00)
To: Rene J Buesa <rjbu...@yahoo.com<mailto:rjbu...@yahoo.com>>, "Vickroy, 
James" <jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>>
Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@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@lists.utsouthwestern.edu]
Sent: Tuesday, February 16, 2016 1:12 PM
To: Vickroy, James; 
histonet@lists.utsouthwestern.edu<mailto:histonet@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.rené

On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
<histonet@lists.utsouthwestern.edu<mailto:histonet@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:  
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[Histonet] nuclear bubbling

[Carlos Defeo via Histonet]

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
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Re: [Histonet] Nuclear Bubbling

[Jamal Rowaihi via Histonet]

Great, I agree 


Regards
Jamal RowaihiAnatomic Pathology SupervisorAl Borg Medical Laboratories Sent 
from my cell phone Original message From: "Manfre, Philip via 
Histonet" <histonet@lists.utsouthwestern.edu> Date: 2/16/2016  10:44 PM  
(GMT+03:00) To: Rene J Buesa <rjbu...@yahoo.com>, "Vickroy, James" 
<jvick...@springfieldclinic.com> Cc: histonet@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@lists.utsouthwestern.edu] 
Sent: Tuesday, February 16, 2016 1:12 PM
To: Vickroy, James; histonet@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.rené 

    On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
<histonet@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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>



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Re: [Histonet] Nuclear Bubbling

[Manfre, Philip via Histonet]

Sort of a rude response to someone looking for help.

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

On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
<histonet@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:  jvick...@springfieldclinic.com<mailto:jvick...@springfieldclinic.com>



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Re: [Histonet] Nuclear Bubbling

[Jennifer MacDonald via Histonet]

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@lists.utsouthwestern.edu>
To: "Vickroy, James" <jvick...@springfieldclinic.com>, 
"histonet@lists.utsouthwestern.edu" <histonet@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.rené 

On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
<histonet@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:  jvick...@springfieldclinic.com<
mailto:jvick...@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. 
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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 
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Re: [Histonet] Nuclear Bubbling

[Rene J Buesa via Histonet]

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.rené 

On Tuesday, February 16, 2016 12:32 PM, "Vickroy, James via Histonet" 
 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:  jvick...@springfieldclinic.com



This electronic message contains information from Springfield Clinic, LLP that 
may be confidential, privileged, and/or sensitive. This information is intended 
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taken on the contents of this information is strictly prohibited. If you have 
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[Histonet] Nuclear Bubbling

[Vickroy, James via Histonet]

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:  jvick...@springfieldclinic.com



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[Histonet] Nuclear bubbling

[Vickroy, James via Histonet]

Traditionally we regard nuclear bubbling as incomplete fixation however I'm not 
so sure that nuclear bubbling can't be caused by additional processing 
problems.  This morning I have some GI biopsies that fixed for nearly 18 hrs 
that have a large amount of nuclear bubbling.   We run the biopsies on a "short 
run".   I am wondering if possibly we are not getting rid of all of the water 
and therefore our dehydration steps are not long enough?

Anybody have an idea besides incomplete fixation?

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:  jvick...@springfieldclinic.com



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Re: [Histonet] Nuclear bubbling

[Rene J Buesa via Histonet]

"Nuclear bubbling", manifested as round unstained areas in the nucleus, is 
caused by incomplete dehydration of the section before staining. There is a 
review on the subject that I cannot find at this moment.René 

On Friday, January 29, 2016 10:42 AM, "Vickroy, James via Histonet" 
 wrote:
 

 
Traditionally we regard nuclear bubbling as incomplete fixation however I'm not 
so sure that nuclear bubbling can't be caused by additional processing 
problems.  This morning I have some GI biopsies that fixed for nearly 18 hrs 
that have a large amount of nuclear bubbling.  We run the biopsies on a "short 
run".  I am wondering if possibly we are not getting rid of all of the water 
and therefore our dehydration steps are not long enough?

Anybody have an idea besides incomplete fixation?

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:  jvick...@springfieldclinic.com



This electronic message contains information from Springfield Clinic, LLP that 
may be confidential, privileged, and/or sensitive. This information is intended 
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taken on the contents of this information is strictly prohibited. If you have 
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Re: [Histonet] Nuclear bubbling artifact

[Gudrun Lang via Histonet]

I've read about a group, that observed living cells during the
fixation-process. They saw bubbling in the first period of contact and
penetration of formaldehyde. After a certain time the bubbles disappeared
again. 
Along this observation for me bubbles are a sign of too short fixation.

Gudrun Lang

-Ursprüngliche Nachricht-
Von: Teri Johnson via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Freitag, 29. Jänner 2016 19:49
An: 'histonet@lists.utsouthwestern.edu'
Betreff: Re: [Histonet] Nuclear bubbling artifact

Hi James,

Nuclear bubbling artifact is most commonly seen in formalin fixed epithelial
cells, and GI biopsies are among those samples that are particularly
susceptible to it. It has been linked to inadequate fixation and also to
heating of slides prior to staining without complete air-drying of the
tissues. I would recommend cutting the block again, air drying the slides
for a time before using heat to melt the wax prior to H stain and see if
the artifact persists.

http://www.cap.org/apps/docs/proficiency_testing/nuclear_bubbling.pdf

Best wishes,
Teri Johnson




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Re: [Histonet] Nuclear bubbling artifact

[Teri Johnson via Histonet]

Hi James,

Nuclear bubbling artifact is most commonly seen in formalin fixed epithelial 
cells, and GI biopsies are among those samples that are particularly 
susceptible to it. It has been linked to inadequate fixation and also to 
heating of slides prior to staining without complete air-drying of the tissues. 
I would recommend cutting the block again, air drying the slides for a time 
before using heat to melt the wax prior to H stain and see if the artifact 
persists.

http://www.cap.org/apps/docs/proficiency_testing/nuclear_bubbling.pdf

Best wishes,
Teri Johnson




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RE: [Histonet] nuclear bubbling

[Patsy Ruegg]

This is most likely caused by inadequate fixation in formalin which would
protect the tissues from tissue processing.  With such fast turn around
times we are seeing this more and more.  Put formalin on your tissue
processor and use any extra time you may have up front in formalin rather
than tissue processing.  Ideally it takes 24 hrs to properly fix tissues so
they are not damaged by processing and embedding in paraffin, but nobody
fixes that long anymore.

Regards,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
 

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-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, May 12, 2011 1:41 PM
To: Johnson, Nacaela; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] nuclear bubbling

Nacaela: 
Heat by itself is not the cause, but the heat applied to sections that have
not been properly drained and still have water left between them and the
slide, is.
You have to drain the sections properly before heating them.
Using formalin is not the cause.
René J.

From: Johnson, Nacaela nacaela.john...@usoncology.com
To: histonet@lists.utsouthwestern.edu
Sent: Thursday, May 12, 2011 12:51 PM
Subject: [Histonet] nuclear bubbling

Has anyone one had problems with nuclear bubbling?  I have read about
two different causes.  (1) Formalin itself is known to cause the issue
and (2) heating the tissue at a high temp (70 degrees C or above) while
drying.  The latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during
collection.  Does anyone have any other suggestions?  Has this problem
occurred for any other reason than I have already pointed out?  It seems
to be worse in the surgical needle biopsies than in the bone marrow
biopsies.


Thanks,



Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com


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[Histonet] nuclear bubbling

[Johnson, Nacaela]

Has anyone one had problems with nuclear bubbling?  I have read about
two different causes.  (1) Formalin itself is known to cause the issue
and (2) heating the tissue at a high temp (70 degrees C or above) while
drying.  The latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during
collection.  Does anyone have any other suggestions?  Has this problem
occurred for any other reason than I have already pointed out?  It seems
to be worse in the surgical needle biopsies than in the bone marrow
biopsies.
 

Thanks,

 

Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com

 
/preThe contents of this electronic mail message and any attachments are 
confidential, possibly privileged and intended for the addressee(s) 
only.brOnly the addressee(s) may read, disseminate, retain or otherwise use 
this message. If received in error, please immediately inform the sender and 
then delete this message without disclosing its contents to anyone./pre
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AW: [Histonet] nuclear bubbling

[Gudrun Lang]

http://decapoda.nhm.org/pdfs/1505/1505.pdf

Here you can find an assay about formalin fixation. On page 851 there is a
description of nuclear and cytoplasmic bubbling found few minutes after
addition of formaldehyd-solution to cultured cells.

My interpretation is, that too short fixation makes the bubbling visible in
the tissue slides. And that after adequate duration of fixation the bubbles
are gone somehow.

What do you think?

Gudrun



-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Johnson,
Nacaela
Gesendet: Donnerstag, 12. Mai 2011 18:51
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] nuclear bubbling

Has anyone one had problems with nuclear bubbling?  I have read about
two different causes.  (1) Formalin itself is known to cause the issue
and (2) heating the tissue at a high temp (70 degrees C or above) while
drying.  The latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during
collection.  Does anyone have any other suggestions?  Has this problem
occurred for any other reason than I have already pointed out?  It seems
to be worse in the surgical needle biopsies than in the bone marrow
biopsies.


Thanks,



Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com


/preThe contents of this electronic mail message and any attachments are
confidential, possibly privileged and intended for the addressee(s)
only.brOnly the addressee(s) may read, disseminate, retain or otherwise
use this message. If received in error, please immediately inform the sender
and then delete this message without disclosing its contents to
anyone./pre
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Re: [Histonet] nuclear bubbling

[Rene J Buesa]

Nacaela: 
Heat by itself is not the cause, but the heat applied to sections that have not 
been properly drained and still have water left between them and the slide, is.
You have to drain the sections properly before heating them.
Using formalin is not the cause.
René J.

From: Johnson, Nacaela nacaela.john...@usoncology.com
To: histonet@lists.utsouthwestern.edu
Sent: Thursday, May 12, 2011 12:51 PM
Subject: [Histonet] nuclear bubbling

Has anyone one had problems with nuclear bubbling?  I have read about
two different causes.  (1) Formalin itself is known to cause the issue
and (2) heating the tissue at a high temp (70 degrees C or above) while
drying.  The latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during
collection.  Does anyone have any other suggestions?  Has this problem
occurred for any other reason than I have already pointed out?  It seems
to be worse in the surgical needle biopsies than in the bone marrow
biopsies.


Thanks,



Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com


/preThe contents of this electronic mail message and any attachments are 
confidential, possibly privileged and intended for the addressee(s) 
only.brOnly the addressee(s) may read, disseminate, retain or otherwise use 
this message. If received in error, please immediately inform the sender and 
then delete this message without disclosing its contents to anyone./pre
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RE: [Histonet] nuclear bubbling

[Johnson, Nacaela]

Another individual called me and commented further, which clarified your 
statement.  It is actually poor lab practice to combine the drying/baking.  
These steps should really be done separately so as to not trap the water 
under the tissue.  I have noticed bubbles of water on the bottom of my slides 
that are encapsulated in wax.  I assume this is also happening under the tissue 
as well, therefore affecting the nuclear morphology.  I had been taught to 
speed up the process by placing them in the oven directly after cutting.  
Decreasing the TAT was all they really cared about.  Fortunately where I work 
now chooses quality over quantity and I can adjust my protocol to get rid of 
the artifact.  Thank you all for your comments and clarity.


Thanks,
 
Nacaela Johnson, B.S. HTL (ASCP)CM
Histotechnologist
KCCC Pathology
12000 110th St., Ste. 400
Overland Park, KS 66210
Office:  913-234-0576
Fax:  913-433-7639
Email:  nacaela.john...@usoncology.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Johnson, Nacaela
Sent: Thursday, May 12, 2011 2:50 PM
To: Rene J Buesa; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] nuclear bubbling

So there really isn't a point in using an oven to dry the slides?  I currently 
dry my slides for an hour at 55-60 degrees C. 


Thanks,
 
Nacaela Johnson, B.S. HTL (ASCP)CM
Histotechnologist
KCCC Pathology
12000 110th St., Ste. 400
Overland Park, KS 66210
Office:  913-234-0576
Fax:  913-433-7639
Email:  nacaela.john...@usoncology.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, May 12, 2011 2:41 PM
To: Johnson, Nacaela; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] nuclear bubbling

Nacaela: 
Heat by itself is not the cause, but the heat applied to sections that have not 
been properly drained and still have water left between them and the slide, is.
You have to drain the sections properly before heating them.
Using formalin is not the cause.
René J.

From: Johnson, Nacaela nacaela.john...@usoncology.com
To: histonet@lists.utsouthwestern.edu
Sent: Thursday, May 12, 2011 12:51 PM
Subject: [Histonet] nuclear bubbling

Has anyone one had problems with nuclear bubbling?  I have read about two 
different causes.  (1) Formalin itself is known to cause the issue and (2) 
heating the tissue at a high temp (70 degrees C or above) while drying.  The 
latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during 
collection.  Does anyone have any other suggestions?  Has this problem occurred 
for any other reason than I have already pointed out?  It seems to be worse in 
the surgical needle biopsies than in the bone marrow biopsies.


Thanks,



Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com


/preThe contents of this electronic mail message and any attachments are 
confidential, possibly privileged and intended for the addressee(s) 
only.brOnly the addressee(s) may read, disseminate, retain or otherwise use 
this message. If received in error, please immediately inform the sender and 
then delete this message without disclosing its contents to anyone./pre 
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RE: [Histonet] nuclear bubbling

[Goins, Tresa]

The best tip I ever received was from a gentleman in Australia who suggested 
adding a very small - so small it would appear to be useless - amount of Tween 
20 to the floatation water bath.  The entrapment of water is eliminated - no 
more blebs of water at the base of the sections.  Works like magic.


Tresa Goins
Histopathology Supervisor
Department of Livestock
Bozeman, Montana

  

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Johnson, Nacaela
Sent: Thursday, May 12, 2011 2:25 PM
To: Rene J Buesa; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] nuclear bubbling

Another individual called me and commented further, which clarified your 
statement.  It is actually poor lab practice to combine the drying/baking.  
These steps should really be done separately so as to not trap the water 
under the tissue.  I have noticed bubbles of water on the bottom of my slides 
that are encapsulated in wax.  I assume this is also happening under the tissue 
as well, therefore affecting the nuclear morphology.  I had been taught to 
speed up the process by placing them in the oven directly after cutting.  
Decreasing the TAT was all they really cared about.  Fortunately where I work 
now chooses quality over quantity and I can adjust my protocol to get rid of 
the artifact.  Thank you all for your comments and clarity.


Thanks,
 
Nacaela Johnson, B.S. HTL (ASCP)CM
Histotechnologist
KCCC Pathology
12000 110th St., Ste. 400
Overland Park, KS 66210
Office:  913-234-0576
Fax:  913-433-7639
Email:  nacaela.john...@usoncology.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Johnson, Nacaela
Sent: Thursday, May 12, 2011 2:50 PM
To: Rene J Buesa; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] nuclear bubbling

So there really isn't a point in using an oven to dry the slides?  I currently 
dry my slides for an hour at 55-60 degrees C. 


Thanks,
 
Nacaela Johnson, B.S. HTL (ASCP)CM
Histotechnologist
KCCC Pathology
12000 110th St., Ste. 400
Overland Park, KS 66210
Office:  913-234-0576
Fax:  913-433-7639
Email:  nacaela.john...@usoncology.com

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, May 12, 2011 2:41 PM
To: Johnson, Nacaela; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] nuclear bubbling

Nacaela: 
Heat by itself is not the cause, but the heat applied to sections that have not 
been properly drained and still have water left between them and the slide, is.
You have to drain the sections properly before heating them.
Using formalin is not the cause.
René J.

From: Johnson, Nacaela nacaela.john...@usoncology.com
To: histonet@lists.utsouthwestern.edu
Sent: Thursday, May 12, 2011 12:51 PM
Subject: [Histonet] nuclear bubbling

Has anyone one had problems with nuclear bubbling?  I have read about two 
different causes.  (1) Formalin itself is known to cause the issue and (2) 
heating the tissue at a high temp (70 degrees C or above) while drying.  The 
latter is definitely not happening, but I do use formalin.
I am in the process of changing the type of formalin that is used during 
collection.  Does anyone have any other suggestions?  Has this problem occurred 
for any other reason than I have already pointed out?  It seems to be worse in 
the surgical needle biopsies than in the bone marrow biopsies.


Thanks,



Nacaela Johnson, B.S. HTL (ASCP)CM

Histotechnologist

KCCC Pathology

12000 110th St., Ste. 400

Overland Park, KS 66210

Office:  913-234-0576

Fax:  913-433-7639

Email:  nacaela.john...@usoncology.com


/preThe contents of this electronic mail message and any attachments are 
confidential, possibly privileged and intended for the addressee(s) 
only.brOnly the addressee(s) may read, disseminate, retain or otherwise use 
this message. If received in error, please immediately inform the sender and 
then delete this message without disclosing its contents to anyone./pre 
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RE: [Histonet] nuclear bubbling

[Rene J Buesa]

Exactly! The sections cannot have any water remaining between them and the 
slide. That is the thing. Microwaving them in that condition (with water 
underneath) will also produce the same artifact.
René J.

--- On Fri, 7/17/09, Harrison, Sandra C. sandra.harris...@va.gov wrote:


From: Harrison, Sandra C. sandra.harris...@va.gov
Subject: RE: [Histonet] nuclear bubbling
To: Histonet histonet@lists.utsouthwestern.edu
Date: Friday, July 17, 2009, 5:27 PM


How about microwaving to dry slides?  Can that cause nuclear bubbling?  

When you say completely drained off, does that mean your slides have to be 
completely dry prior to placing them in the oven?

Thanks,
Sandy

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, July 15, 2009 2:59 PM
To: Histonet; Joyce Cline
Subject: Re: [Histonet] nuclear bubbling

Anybody can experience nuclear bubbling in any type tissue as long as the 
sections as set to dry at high temperature BEFORE they are completely drained 
off!
René J.

--- On Wed, 7/15/09, Joyce Cline jcl...@wchsys.org wrote:


From: Joyce Cline jcl...@wchsys.org
Subject: [Histonet] nuclear bubbling
To: Histonet histonet@lists.utsouthwestern.edu
Date: Wednesday, July 15, 2009, 3:45 PM


Has anyone experienced nuclear bubbling on prostate biopsies? 



Joyce 





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RE: [Histonet] nuclear bubbling

[Maxim Peshkov]

Sandra:
The volume of water under the section onto slide are
very small for microwave heating and this water
cannot be heated adequately. Paraffin is transparent
for microwave and therefore instead drying of slides
you will have a damaged magnetron.
Never dry paraffin sections in MWO!
Maxim Peshkov,
Russia,
Taganrog.

--- On Fri, 7/17/09, Harrison, Sandra C. sandra.harris...@va.gov wrote:


From: Harrison, Sandra C. sandra.harris...@va.gov
Subject: RE: [Histonet] nuclear bubbling
To: Histonet histonet@lists.utsouthwestern.edu
Date: Friday, July 17, 2009, 5:27 PM


How about microwaving to dry slides?
Can that cause nuclear bubbling? 
When you say completely drained off,
does that mean your slides have to be
completely dry prior to placing them in the oven?
Thanks,
Sandy


mailto:maxim...@mail.ru


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RE: [Histonet] nuclear bubbling

[Harrison, Sandra C.]

How about microwaving to dry slides?  Can that cause nuclear bubbling?  

When you say completely drained off, does that mean your slides have to be 
completely dry prior to placing them in the oven?

Thanks,
Sandy

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, July 15, 2009 2:59 PM
To: Histonet; Joyce Cline
Subject: Re: [Histonet] nuclear bubbling

Anybody can experience nuclear bubbling in any type tissue as long as the 
sections as set to dry at high temperature BEFORE they are completely drained 
off!
René J.

--- On Wed, 7/15/09, Joyce Cline jcl...@wchsys.org wrote:


From: Joyce Cline jcl...@wchsys.org
Subject: [Histonet] nuclear bubbling
To: Histonet histonet@lists.utsouthwestern.edu
Date: Wednesday, July 15, 2009, 3:45 PM


Has anyone experienced nuclear bubbling on prostate biopsies? 



Joyce 





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RE: [Histonet] nuclear bubbling

[Kemlo Rogerson]

Sub optimal fixation and as Rene said; drying off at high temperatures. 


 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joyce
Cline
Sent: 15 July 2009 20:45
To: Histonet
Subject: [Histonet] nuclear bubbling

Has anyone experienced nuclear bubbling on prostate biopsies? 

 

Joyce 

 



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[Histonet] nuclear bubbling

[Joyce Cline]

Has anyone experienced nuclear bubbling on prostate biopsies? 

 

Joyce 

 



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Re: [Histonet] nuclear bubbling

[Rene J Buesa]

Anybody can experience nuclear bubbling in any type tissue as long as the 
sections as set to dry at high temperature BEFORE they are completely drained 
off!
René J.

--- On Wed, 7/15/09, Joyce Cline jcl...@wchsys.org wrote:


From: Joyce Cline jcl...@wchsys.org
Subject: [Histonet] nuclear bubbling
To: Histonet histonet@lists.utsouthwestern.edu
Date: Wednesday, July 15, 2009, 3:45 PM


Has anyone experienced nuclear bubbling on prostate biopsies? 



Joyce 





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