>Dear Listmembers,
>
>I am currently using a Zeiss LSM510 with an inverted Axiovert 100 M, as well
>as an CCD imaging system build around an Axiovert S100 TV.
>Both systems I use to measure physiological ion concentrations over longer
>periods of time (10-30min), but I have noticed that with both microscopes
>the focus isn't stable and is completely lost over the course of an
>experiment. With the confocal it's about 5mum in 10min, going downwards.
>Since the cells are only max 5mum thick, this is intolerable. Initially I
>thought that on the imaging system the focus instability might be due to a
>weak table that responds to weight differences during perfusion. But even
>without perfusion, as well as on the confocal with a much more stable table
>the focus drifts. I am still waiting for a detailed response from Zeiss, but
>the first answer was that this seems to be an hitherto unknown problem. So
>my question is:
>
>Has anybody experienced similar problems with focus stability on an Axiovert
>100?
>
>Or is it bad coincidence that both my microscopes happen to be faulty and I
>just have to buy two "greatly improved Axiovert 200 with maximum
>stability"...
>
>Cheers
>
>Frank

Dear Frank,

Yes, we have experienced focus shift problems on our Axiovert 100
microscopes, but it turned out to be caused by temperature fluctuations. We
now have each microscope enclosed within a Perspex incubation chamber, and
the focus remains stable. The box is designed so that it has a laminar flow
system for the circulating warm air,and the temperature is accurately
controlled. It's important that the whole of the microscope body is
enclosed within the box!

Of course you may have an entirely different problem.

Hope this helps

Peter Jordan

>
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Peter Jordan
Light Microscopy
Room 1B11
Imperial Cancer Research Fund
44, Lincoln's Inn Fields
London
WC2A 3PX

Tel. 0207 269 3564
Fax 0207 269 2859