[Histonet] Ventana Retic problems

[Jeffrey Robinson]

Hi Donna-  I have been dealing with these same exact issues for 3 1/2 years now so I think I have achieved "expert status" in dealing with this problem.

First, I'll give you my current protocol.  When all things are working smoothly, it produces a nice stain.  But it is very frustrating when it starts to "fade."

We cut all retics at 5.  Thinner cuts will definitely look lighter.

Protocol:  warmup slide:  47 degrees C.
             Oxidizer:    4 minutes
             Decolorizer:         4 minutes
             Sensitizer:          8 minutes
             Optimize Counterstain Intensity:  4 Minutes

I have gone around and around with Ventana on decontamination problems with this instrument.  I was performing full decontamination runs every month.  My rep finally said to just decon the bulk wash carboy and the wash bottle on the instrument when fading begins to show.  Ventana claims there will be a new wash out this year that will hopefully take care of the problem once and for all but no one at Ventana can give me a release date on that.
In the meantime, this is what I do:

Daily:  run the "Purge Wash" function test 3 times in the morning before running any slides.  Be sure to run the "Purge" function and not the "Prime" function.
When loading slides, put all of your retic slides on after everything else (after the GMS, etc., slides).

When staining starts to fade:  after ruling out "thin cuts" and other obvious problems it is time for decon.  The fading will show first on the patient tissue (the control may still look OK).
Here is my current decon protocol:  I use the Lysol IC decon solution. I have an extra 6 gallon carboy and I leave some made up (diluted) so that it is ready to go.  Put some decon solution on a 4X4 guaze and wipe the dispenser tip underneath the top (lift lid up for access).  Dump the wash solution in the wash bottle on the instrument.  Rinse out with DI water. Fill with decon solution.  Swish solution inside bottle.  Replace bottle (on instrument).  Run "Purge Wash" function test (3 times).  After Purge #3, let the solution sit in the lines for at least 15 minutes (the longer the better).  When you are ready to proceed, rinse bulk bottle well several times and replace with DI water.  Run "Purge Wash" 3 more times.  Time is not a factor here so you can just run them one after another.  After the third purge, replace the DI with BM SS wash.  Run "Purge Wash" function test 3 more times.  That's it.  I find I need to run this procedure about once a month.
Additionally, I decon the 5 gallon bulk wash carboy EVERY TIME I make up a new batch.  It doesn't take long (I just let the decon solution sit for 15 minutes) and then when I do need to do the modified decon on the instrument I do not need to worry about the bulk carboy.

I hope this helps- it takes a little time but it does seems to help with the retic stain intensity.

Jeff Robinson, Senior Histotechnologist (HT, HTL), Sierra Pathology Lab, Clovis, CA.

-----Original Message-----
From: donna mihalik rossi via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Wednesday, June 14, 2017 1:45 PM
To: histonet
Subject: [Histonet] Ventana Retic problems

Hi Histonetters!  We are experiencing sporadic staining of retic fibers from our Ventana Benchmark machines.  We have 2 machines and are having the same problem on both. The machines were both decontaminated 2 weeks ago with all new solutions  being made.  The fibers are not crisp and are disconnected.  Our control tissue is  at the top of the slide with the patient tissue at the bottom. It appears that the control is staining better than the patient tissue but still is  not crisp. We have tried different lots with the same result. Any comments before we consult Ventana? Your help would be appreciated.  Thanks, Donna  Rossi, PSU


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