I'm not certain that this will fix your problem, but I'd
ditch the Tween in the final wash or two before mounting.
There may be too much residual buffer on your slides when
you add the Prolong - I wick off as much as I can, without
drying the tissue, before mounting.
You let the Prolong cure overnight - so why the nail
polish? The solvents in nail polish can cause bleaching
&/or "fuzziness" in the background & you don't need it if
you are using a curing mount. If your slides aren't cured
after an overnight "sit", you may just be using wayyy too
much mounting media.
As to why the problem has suddenly started cropping
up....anybody's guess. Bad karma?
Tamara
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Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
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