Reply from a vendor:

In a message dated 11/1/97 5:39:53, [log in to unmask] (Barbara
Tolloczko) wrote:

<<We are imaging [Ca2+}i changes in living cells (loded with
Fura-2) using PTI system (hardware and software), based on a Nikon
microscope, and CCD camera with intensifier from Videoscope. This is
good enough to see changes in individual cells, but now we would like to
be able to moniter changes in subcellular compartments. What would we
need to upgrade our system (camera, deconvolution software, image
analysis software etc) and what would be the cost of it? Or would it
be better to go for a real time confocal microscopy? Or two photon?
(I am allowed to make "a dream list").>>

You could add confocal capability to your current set-up with an InSight
Basic.  This puts a video rate confocal scanner between your oculars and the
microscope body and illuminates the sample with an argon ion laser.  You can
then use the camera you have, the intesifier you have, and the software you
know to collect and analyze your data.  Options include Krypton laser, fast
shutters, Z-drive, filter wheel, IQ computer, and cooled-intensified CCD
camera, depending on how big a dream list you can get away with.

You would not be able to excite FURA-2 with the laser, although the confocal
aperture will give you some benefit on resolution.  Better to use Fura Red or
Fluo-3 or a combination of the two.  There are many other blue excited dyes
out there, and the choice depends on the experiment.

If you have the budget, 2-photon would be an option.  If you have the time,
deconvolution may also help, although an intensifier placed in series may
insert too much noise and remove too much dynamic range to give you much
improvement.

David
Biovision (formerly Meridian)

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