Posted by
Christian Goosmann-2 on
Mar 27, 2013; 9:44am
URL: http://imagej.273.s1.nabble.com/GUS-staining-quantification-tp5002411p5002452.html
Dear Magi,
in a brightfield image you are looking at densities not at intensities.
What you tried would be ok for fluorescence images where a present stain
(dye) produces a proportional intensity of blue light.
When you look at stains in a brightfield image you actually see the
highest intensity in all channels where you see white. Where your blue
stain is present, you see the same intensity in blue but less intensity
in the other channels mainly in red. So the information is indeed in the
red channel and it is inverted because the highest density of your dye
subtracts the most intensty out of your red channel. So if you want a
postive relation to your dye you need to invert the image and look at
the red channel. In a brightfield image you are looking at densities not
at intensities. I am sure there are functions in ImageJ that handle
densities rather than intensities. Look at functions like color
deconvolution etc. I am not too experienced with myself.
Just hope to point you in the right direction.
Christian
---
Christian Goosmann
Mikroskopie
Max-Planck-Institut für Infektionsbiologie
Campus Charité Mitte
Charitéplatz 1
10117 Berlin
Tel.: +49 30 28460 388
magi78343 wrote:
> Dear all,
>
> I need to quantify the GUS-staining intensity (blue) of some digital images.
> As far as I know I first need to split the RGB channels. I am interested
> only in the blue intensity but the blue channel is the pilest one and the
> dark gray color corresponding to the dark blue on the color image remains in
> the red channel. Which channel should I use for measurement?
>
> Thank you in advance.
>
> Magi
>
>
>
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