Re: [Histonet] Immuno slide problem
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Short answer, slides sound suspect. More elaboration below ****.
From: "Anissa Choi"
Thanks to all who replied to my question. Please see if the following
will help to gain new insight into my problem.
2.The problem is not repeating. We recut using slides from the same
batch and everything was fine. The other day, out of a run of 20 slides,
2 slides did not work( both control and test did not work). We checked
dispenser, AEC kit etc and could not find anything wrong. We repeated
without altering anything, same control, same AEC kit, same protease,
same antibody dispenser and slides from same batch. The repeated run was
****Assuming you had the same number of slides loaded, i.e. a bad slot
(or slots) on the machine was not excluded from the follow-up run, it
sounds like a bad batch of slides.
3. Ventana suggested the red paint may get onto some slides and causes
patchy and uneven staining. I was told that the particles from the red
stuff affect the staining reaction on the Ventana stainer.
****Its inconsistency would refute this argument if it was exclusively
the red "paint" that was hindering the staining. Conceivably, Ventana's
hypothesis is possible but the intermittent irregularity seems weird and
inconclusive. May be a sealant or "post-painting" process in the
manufacture of these slides that is leaving trace chemicals - so it's
not the paint but something else coating the slides that is creating
"non-stain zones", for lack of a better description.
5. Is it reasonable to conclude that we have a slide problem (occasional
defective slides in the batch), rather than a machine problem ? Would
you switch to other type of charged slides?
****I would try several brands of charged slides and run them
concurrently with only positive and negative controls. Forget real
specimens until you get this resolved. Record the slot number of
positives that do not stain (assuming some of them do not stain). You
may even need to try a very common, universal Ab that detects a very
common antigen present in all tissues to ensure that your Ab doesn't
introduce another set of issues. If another batch of slides results in
this irregular pattern, then it almost has to be the Ventana.
Hope this helps,
"Biology in a new form..."
Member Services: www.myhistosoft.com
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