Posted by
Glen MacDonald-2 on
Nov 05, 2015; 10:25pm
URL: http://imagej.273.s1.nabble.com/Problem-with-channel-bleed-through-after-splitting-channels-from-RGB-image-tp5014842p5014868.html
Dear FrankyG,
You will need to get the specifications for your particular Evos. They come with different configurations and different cameras. You should also have the filter specs available. Your comments suggest 2 cameras on your Evos? If you have a trully monochrome camera, then it should be used for quantitative imaging, collecting each channel sequentially. If you have a CCD with mosaic filters (Bayer mask) there is a modest loss of spatial resolution, overlap between the camera filters means any bleedthrough in the imaging system will be picked up by the other channels, you are limited to 8-bits/pixel and unless you have access to the RAW format, the “colors’ are established by firmware interpolation of intensity neighborhoods. An RGB image from a Bayer mask color camera can be set to gray, but its just a LUT manipulation and you still have the above issues. A monochrome camerea does not need to save as RAW since there is no need to alter the captured intensities, provided you do save it in full bit depth rather than as RGB. However, I haven’t used those scopes in a long time and don’t know what they are up these days with their software.
Regards
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Cellular Morphology Core
Center on Human Development and Disability
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[hidden email]
On Nov 5, 2015, at 1:34 PM, FrankyG <
[hidden email]> wrote:
> Dr. Landini,
>
> I save all of them in .tif.
>
> I'm not sure what artifacts you are referring to and I stopped using the RGB
> for quantitative analysis because of the issues I realized once I split the
> images. Saving them as separate channels fixes almost all the issues I have
> been having. Perhaps look at the images that are separate channels and see
> if you can find artifacts? Those are the ones I am using now, not the RGB.
>
> To be clear they are saved separately from the RGB, perhaps that was not
> clear.
>
> Dr. MacDonald,
>
> Yes our EVOS has a monochrome camera and the representative explained it to
> me. I'm willing to bet it is an RGB camera as well as I cannot save as RAW,
> only RGB format.
>
> So are you saying that even if I save the channels separately that the
> imaging system I am using is absolutely useless for quantitative studies? If
> so I'll have to notify my PI ASAP because I do not think he was aware of
> that.
>
>
>
> --
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