[Histonet] Rapid tissue Programs

Tue Nov 24 12:20:09 CST 2015

You point out to several issues that I would like to address:1- "nuclear bubbling" has nothing to do with processing. This is a post-sectioning artifact appearing when there is water underneath the section when it is set to dry before staining. Just make sure you shake the slide with the section and set it to drain vertically before placing them into the oven to dry.2- sponges can be a source of problems influencing dehydration and even causing "marks" on the tissue that can later produce an artifact. Try to avoid sponges and use tissue paper to wrap the biopsies instead.3- fear not placing small biopsies in the overnight "long process". If the protocol is well balances you will have no problems. In all the years I oversaw tissue processing ine 2 labs I supervised (one with an excess of 35,000 cases/years and the other close to 200,000 cases/year) I had only one protocol for everything, except for fatty tissues, breast and brain.The time in every reagent is not really the issue but the gradient you use. Abrupt changes (like starting with 90% alcohols) or not having "mixed steps" (1:1 ratios) between last alcohol and clearing agent or between clearing agent and melted paraffin are the real causes of the so called "harsh" processing.
If you go to http://www.histosearch.com/rene/html you will find my "standard" protocols.René 

    On Tuesday, November 24, 2015 12:24 PM, "Vickroy, James via Histonet" <histonet at lists.utsouthwestern.edu> wrote:

 Our latest CAP survey was returned today and although there are no major issues one possible improvement area is evident.  On all of the biopsies the area of "fixation/processing" was not rated as excellent.  The suggested possible reasons were:  fixation incomplete, nuclear bubbling artifact, tissue poorly processed.  The survey also said that many of the samples sent in throughout the country had similar  issues in the fixation/processing area most likely because of the rapid turnaround times and shortened processing times.  I am trying to be proactive here and see if we can adjust some times to improve the processing quality even though we have not had any complaints from the pathologists.  Of course we all know that other artifacts caused prior to the specimen arriving in the lab can also have an effect on the quality of the H&E slides.  Our fixation times should not be a factor so I have to conclude that maybe the rest of our processing times need to be adjusted.  Another factor that we have is that we use blue sponges for almost all of our tissues.  Our largest number of specimens are GI biopsies.    If possible can anyone share with me their rapid processing schedules or simply the approximate times they have for each dehydration or clearing step. We do run a larger overnight tissue run on any biopsy or tissue that we feel is too large for the "rapid run".    I am hesitant to run the biopsies routinely on the longer programs becase of over dehydration, etc. even though we do use an alcohol blend.

Any suggestions or similar experiences please share.    Again our pathologists say everything looks great so I don't want to change much.

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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