Shandon Coverplate IHC staining

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From:Gayle Callis <>
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The rinsing with these is something one needs to optimize for his/her
laboratory, one rinse for 4-5 min was adequate for our murine CD markers,
frozen, and out staining was clean and crisp with a full load of buffer in
the coverplate well.

I know of several people who have used the one rinse method, between
applications, and the Shandon brochure on this shows 2 ml of buffer (mine
was closer to 5 mls, whatever filled the well) and they did no indicate the
number of rinses.  

Caveat:  When I did more than one rinse, with a full well, the buffer waste
in the bottom of the tray (reagent waste) was raised dangerously close to
the BOTTOM of the slides, and fear of backwashing into the gap led to
aspiration of the buffer OUT of this tray via an empty slide/coverplate
slot. So be careful with lots of rinses, you could "back up the plumbing" ,
particularly when you fill the tray with all 10 coverplates.  

One other thing, has anyone ever calculated the thickness of the little
plus charge markers on the bottom of the Erie Plus charge slides?  Did that
when curiosity and use the coverplate system forced this issue here, Erie
nicely supplied the thickness.   When you put a coverplate up against one
of these slides, you have a bigger gap than you think, in fact,
approximately 140 um! which is keeping your slide surface AWAY from the
flat coverplate.  The buffer flow much faster than with slide without these
little 'pluses', so take that into account on your timing for a rinse. 

Forever scientifically curious! or nit-picking!

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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