>Dear listmembers,
>
>I have a problem that pops up while doing confocal physiology
>experiments.
>I use a 63x water immersion lens (because the cells are 30um thick) on
>an inverted Leica TCS-SP. I  image these cells for extended periods of
>time (1-2 hours, but I plan to go up to 48 hours) at 37 degrees. Both
>the culture chamber and the objective are temperature controlled. The
>problem: within 30 minutes, the water droplet on the lens has
>evaporated.
>
>Has anyone solved this problem, or ideas how to proceed?

Here's a low-cost solution: Make an angled micropipet that, when
strapped to the objective barrel, has its  tip just contacting the
immersion droplet. Supply (filtered) water, i.e. by gravity feed.
It'll probably take some fiddling until you get micropipets with a
suitable shape and opening (although it should be easy for a proven
electrophysiologist  ;-) ) and the right pressure/height to replace
the water at the same rate it evaporates.
Make sure the pipet tip ends outside the front lens area or you might
get a reversible optical distortion immediately - and an irreversible
one when you crush the tip between the front lens and the coverslip.

>  Does immersion
>oil with same index as water exist?

I don't think so. But let me know if you find one.

Cheers,

       Beat


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