[Histonet] Nuclear Bubbling

[Manfre, Philip]

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.rené 

    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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