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Chris Tully <[log in to unmask]> writes:

> well plates perfect focus systems can be VERY useful.  While taking a z stack
> can help, I have seen more than one multi well plate with enough variation
> from well to well (especially from one end of the plate to the other) to

One important factor is to have a setup that allows position
definition in 3D, ie X,Y and Z. This saves problems from tilted, or
non-flat plates. I have done this with 72h time courses on 24 well
plates without any automatic focus adjustment, but with computer
control of x,y and z to visit the defined positions in 3D. With good
temperature stability this is not difficult at relatively low mag, say
10-20x. At high mag, or more importantly high NA, your depth of focus
is much smaller and therefore this is much more demanding.

Ian