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Eric Olson <[log in to unmask]> writes:

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>
> We study cellular migration and are hoping to perform timelapse imaging
> on multiple wells of a 96 well plate.  The images would be acquired at
> 5 min intervals for each well. Right now we are performing this
> experiment one well at a time so any increase in the number of imaged
> wells would be great.
>
> I am concerned about the precision of the automated XY stages - whether
> the reacquired field will be in register with the previous image.
> Also, I am wondering how people handle the difference in focal plane
> between wells due to minor imprecisions in the manufacture of 96 well
> plate.
> Does this imprecision in the Z axis require that  Z series be performed
> on each well to ensure that the in focus image is captured?  Or can a Z
> coordinate be specified for each well?
>
> Any input on the best approach to multiwell timelapse imaging would be
> appreciated.  Also any information on turnkey systems suitable
> for this application would be useful.

I have done this, and found it very effective. Focus drift due to
thermal effects can be a significant problem. My experiments were
carried out with the microscope enclosed in a heated box, with a PID
controller to maintain a constant temperature. By setting up the
plate, microscope etc, then waiting 60-90 minutes I found stability
was not a problem. A Z-drive was used to get different focus
positions in different wells. The whole system was run from a Kinetic
Imaging system ( http://www.kineticimaging.com/ ). The stage had some
random drift, between time points which was corrected by image
registration, post acquisition.

If you want any more details drop me a line.

Ian