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Alterpauline yu wrote:

> Ms. Foster and Ms. Puhalla bring up two good points about calibration that I've been wondering about myself. While using a stage micrometer is the best way to calibrate measurements of objects in brightfield, how can one do this for confocal? I don't know of any "fluorescent micrometers" and I'm not sure how I would capture an image through the detector in brightfield.

In my experience, you can see the gratings on a stage micrometer using
confocal optics.  I have NO idea of the optical principles behind
this--it might be reflected light bleeding through the barrier filters
or fluorescent oil embedded on the surface.  In fact, I suppose that if
you had trouble seeing the gratings you might try smearing them with a
bit of fluorescent antibody.

Good luck!

Martin Wessendorf



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