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I have a question regarding the use of a mirror for measuring the axial resolution of a confocal imaging system (laser scanning or spinning disk). 

Jim Pawley's handbook has a chapter that describes a few useful tests that can be done across different wavelengths to assess your microscope's axial resolution using a mirror. I have wondered for quite some time if there is any difference between using a mirror and the excitation wavelengths or a fluorescent object (microsphere) to test axial resolution. I could only find one posting on this in the archive:

http://lists.umn.edu/cgi-bin/wa?A2=ind0106&L=CONFOCALMICROSCOPY&D=0&P=8438

So I'm curious, what is the consensus out there right now? Is there a difference or not? I haven't actually had a chance to test it yet and see if the results obtained with a mirror (with laser illumination) match those obtained with a sub-diffraction sized fluorescent microbead.

John Oreopoulos
Research Assistant
Spectral Applied Research
Richmond Hill, Ontario
Canada
www.spectral.ca