[Histonet] Retics on Ventana Benchmark Special Stainer

[Jeffrey Robinson]

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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