Re: Problem with channel bleed-through after splitting channels from RGB image

Posted by Gabriel Landini on Nov 05, 2015; 3:38pm
URL: http://imagej.273.s1.nabble.com/Problem-with-channel-bleed-through-after-splitting-channels-from-RGB-image-tp5014842p5014858.html

On Thursday 05 Nov 2015 07:18:21 FrankyG wrote:
> Dr. Sheffield,
>
> Our scope is a bit different in that for each channel a different light
> source is used. It still filters the light a bit but the DAPI light source
> is different than the GFP light source. Unless I understood you wrong and
> you were not saying that the cubes all filter from one main light source.
>
> I will try your method of saving and then merging the files again. Though it
> requires more macros I'm all for efficient workflow so it will get done

I think there is a much more important problem here. Load your original RGB
image of the blue nuclei into IJ and convert it to a RGB stack.

Then use the B&C applet to increase the contrast of the channels. You will
realise that there is a large number of serious artifacts in your image.
Not sure what that is, perhaps you saved the file in jpeg format first (you
should never use the jpeg format for any serious imaging).

Perhaps you need to recapture the images and make sure that you are saving in
a non-lossy format.

Regards

Gabriel

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