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