Good Morning to all Histotechs- There have been many questions over the past
several months regarding poor retic staining on the Ventana Benchmark Special
Stainer. With the help of my histotech friends, I have now achieved successful
staining on both liver and bone marrow specimens utilizing this instrument.
My primary source of protocol information has been Heather Nowacki at the Mayo
Clinic. Heather says that they use B-5 fixative and RDO Gold decalcifier on
their bone marrow cores. We use B Plus fixative and Immunocal decalcifier so
the choice of an appropriate fixative and decalcifier does not seem to be a
concern in the ability of this instrument to achieve the desired retic staining
intensity.
Here is Heather's protocol for retic: Heat: 45 degrees C.
Oxidizer: 4 minutes
Decolorizer: 4 minutes
Sensitizer: 8 minutes
Silver: 12 minutes
Nuclear Fast Red Counterstain: 4 minutes
I did not need to modify any of Heather's protocol options. I had previously
been unable to achieve any satisfactory staining on bone marrow retic staining
on the Benchmark Special Stainer. I showed my bone marrow retic test slides to
one of my pathologists and he rated them comparable to the retic slides we are
currently staining on the old Ventana Nexes stainer.
Notes: It seems that the primary issue affecting proper retic staining on the
Benchmark Special Stainer has been the lack of sufficient heat applied to the
slides during the staining run. Ventana has released a software upgrade that
allows for a much wider range of heating temperature options during retic
staining. You will need to have the new software upgrade installed by Ventana
before you will be able to use this protocol. Ventana also discussed an
enhanced decontamination protocol for the Benchmark Special Stainer that
requires some additional reagents but the Ventana technical rep who installed
my upgrade determined that the enhanced decontamination protocol was not
necessary in our lab. She felt that this enhanced protocol would only be
beneficial in labs that have had major contamination issues (including Special
Stains Wash pH changes) in the past. We cut all of our retic slides at 5 (even
for the Nexes) and I have been quite happy with the staining intensity on slides
cut at that thickness. Slides cut at 3 or 4 stain too light for my taste.
Acknowledgements: I would like to thank Tim Morken for spotting the Mayo
Clinic poster on Ventana Benchmark Special Stainer Retic staining at NSH last
month. I would also like to thank the team at the Mayo Clinic who worked on
this project allowing the rest of us in the field to benefit from their hard
work. The Mayo team includes: Heather Nowacki, HTL; Mark Keith, HTL; Scott
Antilla, HTL; and Joaquin Garcia, M.D.
I hope this information is of use to my fellow histotechs who have the Ventana
Benchmark Special Stainer in their labs.
Jeff Robinson, HT, HTL, Senior Histotechnologist, Sierra Pathology Lab, Clovis,
CA.
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