Re: Flat-field correction of fluorescent images

Posted by Gabriel Landini on
URL: http://imagej.273.s1.nabble.com/Flat-field-correction-of-fluorescent-images-tp5000936p5000969.html

> I shut off the led light source, I switch off all the lights in the room and
> then capture 10 images for each gain and exposure setting. I thought the
> 4092 is from hot-pixels.  

Most microscopes have a manual shutter that prevents any light reaching the
camera. That avoids switching on and off the light.

Did you take a look at the distribution (histogram) of hot pixels?
If you get such a high number for dark current I wonder if the gain or range
in your camera is set too high. Or maybe the camera is set to auto-gain (some
have it), in which case should be switched off. What camera is this?
Dark current is usually very small unless you integrate for long times.

Cheers

Gabriel

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