Posted by
Glen MacDonald-2 on
Oct 21, 2015; 4:21pm
URL: http://imagej.273.s1.nabble.com/Cyan-colour-instead-of-Blue-how-do-I-convert-it-into-blue-tp5014698p5014699.html
Dear Apo_99,
It sounds as though you converted your confocal images to RGB. The RGB color scheme is just that: red, green and blue
Cyan is the result of adding blue and green. Overlapping pixels in the blue channel were assigned a similar intensity in the green channel. You might be able to accomplish removal by channel subtraction but the result may be artifactual. The best option is to start over and set the source image to blue and green. If you want separate channels for analysis, it is usually best to remain in the higher bit-depth of the original images rather than immediately converting to RGB. Visualize the original files in ImageJ as ‘Composite’ images, where the Channels Tools will allow viewing in any color combination you wish without losing bit depth or mixing channels. Then you can convert to a desired RGB combination. BTW, cyan is much easier to visualize by the human eye and is often used in publications.
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 Oct 21, 2015, at 8:07 AM, apo_99 <
[hidden email]> wrote:
> Dearest members,
>
> I take pictures using a Leica Confocal, staining my samples with blue (DAPI)
> and green (Bodipy) fluorochromes. After applying the different excitation
> and emission channels for the two fluorochromes (in sequential scan mode), I
> obtain the pictures. However this time, instead of using "Blue" colour to
> depict my blue-emission channel, I chose Cyan colour (thus the nucleus
> becomes better visible to the naked eye) for the same (blue) emission range.
>
> Now, normally in the imageJ I split the 3 colour channels of my obtained
> pictures and then I count particles. In this way, I can measure the size of
> the nucleus (blue fluorochrome, blue channel) and my lipids (green
> fluorochrome, green channel).
>
> BUT in the pictures I obtained choosing the cyan colour for the blue
> fluorochrome, the ImageJ splits my channels in the blue one (showing only
> the DAPI-blue nucleus) and in the green channel which shows BOTH the green
> stained lipids AND the blue stained nucleus!
>
> Is there any way to get rid of the blue stained nucleus in my green channel
> pictures? Any answer would be highly appreciated... Thanks!!!
>
> /Apo_99
> Karolinska Institutet,
> Stockholm Sweden
>
>
>
> --
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>
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