Cyan colour instead of Blue - how do I convert it into blue??

Posted by apo_99 on Oct 21, 2015; 3:07pm
URL: http://imagej.273.s1.nabble.com/Cyan-colour-instead-of-Blue-how-do-I-convert-it-into-blue-tp5014698.html

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