>  >Any insights are welcome-
>
>Finding the correct answer would be very interesting, but from the point of
>view of providing imaging services to biologists who are not imaging with
>reflectance, the question we need to answer is how the instrument behaves
>for fluorescent microscopy typically in aqueous media.  For this, we need
>to look at the PSFs (beads or punctate staining) in cell culture media and
>mounting media.  And that the machine behaves for IRM.
>
>Our PSFs with both the Olympus and Nikon infinity corrected objectives with
>the Biorad Radiance 2000 tend to have a cusp on the side away from the
>objective.  But we haven't yet looked at our new water immersion objectives
>in detail.
>


Such a "cusp" is an indication of spherical aberration.  Indeed, any
asymmetry in z-response to a horiz plane or a point object is an
indication of SA. Possible problems with water lenses include the
coverslip being more than about 2 micrometers (yes 2) away from the
thickness the lens was designed for (or the collar set for), using
the wrong oil or the wrong temperature. Indeed, there is USUALLY some
SA present when real people do real confocal.

Cheer,

Jim P.
At the 3D Live-Cell Course, where to our great surprise, it is sunny
for the 4th straight day.  Will wonders never cease?


--
               ****************************************
Prof. James B. Pawley,                             Ph.  608-263-3147
Room 223, Zoology Research Building,               FAX  608-265-5315
1117 Johnson Ave., Madison, WI, 53706  [log in to unmask]
"A scientist is not one who can answer questions
but one who can question answers."
                Theodore Schick Jr., Skeptical Enquirer, 21-2:39