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    1. COHi guys, Thanks already for the replies. I think I should clarify a bit: The images are recorded on the same physical setup and the photon counter that is used as a detector is also fixed. The only variables are the tow laser beams I use to illuminate the sample. The should illuminate exactly the same volume of the sample and for this purpose need to be aligned using optics. The way to check this is to record the image using the the two lasers separately and somehow combine them. One laser should give a much higher resolution than the other... I was thinking to normalize both images.
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    2. COIf I normalize each one in a different color space, for example green and red and then add the images... I can expect to see yellow in the regions of overlap. If the little yellow discs are in the center of the big ones of the other channel, then both lasers are aligned. See this Screenshot: http://picasaweb.google.com/lh/photo/gQBuodNtXFIy8BniPpLtjw?feat=directlink I saw a similar implementation in a commercial package but I don't know enough GDI to handle the normalisation properly. So if someone can help with that I think my problem is solved!
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    3. COJust wanted to add that I really need to normalize the images over either a red or a green channel space. Just extracting Red or Green from either images and adding those doesn't give nice results. I will probably also need some form of thresholding for my images since they can be noisy.
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