Re: An image analysis question

From: Robert L. Becker (rlb@cpt.afip.org)
Date: Wed Jun 07 2000 - 09:44:58 EST


Need to know more about your expts. Are you using arc lamp illumination?
Light intensity delivered to the specimen can vary significantly and
suddenly/periodically with old Hg lamps. We've speculated this is due to
sudden and intermittent (approx every minute or so) shifts in the arc
path. In any case, the problem goes away with installing a new lamp.

I've seen problems with one high-end (cooled CCD, research grade) camera
in which image intensity was reproducibly irregular. That is, at certain
camera exposure settings (and not weird ones, at that), the distribution
of exposure-to-exposure image intensities (determined for each exposure by
averaging the pixel values in each color channel) was bimodal with peaks
differing by up to 10%. A real headache to document, but the
reproducibility according to exposure setting convinced me the problem is
with the camera rather than the light source.

If you need to distinguish intermittent (up/down) variations in light
intensity from a sensor glitch, consider a colleague's suggestion to rig a
way to run light from a battery-powered source through to your sensor.
Depending on your set up, this might get a definite answer more easily
than setting up power monitors, surge isolaters, etc.

Robert L. Becker, Jr.
Col, USAF, MC
Department of Cellular Pathology
Armed Forces Institute of Pathology
Washington, DC 20306-6000
202-782-1573


On 6 Jun 2000, Maris Handley wrote:

>
> Hello Everyone,
>
> We are doing time lapse experiments with a Nikon microscope (using IPLab software).
> When we play the movie back we are seeing large changes in light intensity (almost
> random...not getting brighter continuously or dimmer continuously).  Our exposure time
> isn't being changed.  I believe this is a problem with the light source stability,
> but is there something else that might be causing this?  Has anyone else seen this,
> and what did you do to solve the problem?
>
> Thank you for any help,
> Maris
>
>
> Maris Handley
> Dana-Farber Cancer Institute
> Boston, MA 02115
> (617)632-3179
>



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