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We do this routinely for cells in glass bottom dishes fixed in situ.  We make up the PPD in buffer shortly before use.  The only issue is that the PPD will oxidize with time and the solution will turn orange.  We have not had that affect imaging in relatively short term situations (several days in fridge).  It is not as high resolution since the refractive index does not match glass or oil, but that depends on how critical your imaging needs are.  Dave

On Mar 11, 2013, at 6:56 PM, Kelvin Poon wrote:

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Hi all,

Just wondering if anyone has had much experience with using
p-Phenylenediamine (PPD) as an anti-fade reagent? Rather than incorporate it
into a mounting medium, I wish to use it in a different way:

I have a fixed tissue slice/section that I wish to image WITHOUT a coverslip
using a water immersion lens. Would it be possible to make up some PPD and
add it to the PBS that I will then immerse the sample in for imaging? Most
PPD protocols I came across instruct that the it be made up in glycerol but
I assume that is purely so you can use it directly as a mounting medium.

So to reiterate: Can I make up PPD in PBS (no glycerol) and then use it as
my immersion solution to image a NON-coverslipped tissue slice/section?

Cheers,
Kelvin

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)