Overlay with pseudocolor image

[Sofia Domingues]

Hi,

I would like to overlay two images in ImageJ and failed up to now. One image
is a grayscale image from the brightfield acquisition. However, the other
one is a pseudocolor image, derived from a ratiometric CFP/YFP FRET image
acquisition. Neither in Metamorph nor in ImageJ I was able to overlay these
two images up to now as either the pseudocolor image can not be selected at
all or the information of the pseudocolor gets lost as the image got
flattened to a single color ...

Thanks a lot for your help!
Best, Sofia

[Johannes Schindelin]

Hi,

Post by Sofia Domingues
I would like to overlay two images in ImageJ and failed up to now. One
image is a grayscale image from the brightfield acquisition. However,
the other one is a pseudocolor image, derived from a ratiometric CFP/YFP
FRET image acquisition. Neither in Metamorph nor in ImageJ I was able to
overlay these two images up to now as either the pseudocolor image can
not be selected at all or the information of the pseudocolor gets lost
as the image got flattened to a single color ...

If I were to overlay a gray image with a pseudocolor image, I would
convert both to RGB color and then use Process>Image Calculator to add
them.

But I guess I did not understand what you mean by "or the information of
the pseudocolor gets lost as the image got flattened to a single color".
Maybe you want to be a little more verbose about it, so I have a chance to
understand what you need?

Ciao,
Dscho

[Alicia Piera Alberola]

Post by Johannes Schindelin
Hi,

Post by Sofia Domingues
I would like to overlay two images in ImageJ and failed up to now. One
image is a grayscale image from the brightfield acquisition. However,
the other one is a pseudocolor image, derived from a ratiometric CFP/YFP
FRET image acquisition. Neither in Metamorph nor in ImageJ I was able to
overlay these two images up to now as either the pseudocolor image can
not be selected at all or the information of the pseudocolor gets lost
as the image got flattened to a single color ...

If I were to overlay a gray image with a pseudocolor image, I would
convert both to RGB color and then use Process>Image Calculator to add
them.

If I would like to overlay a gray image with a pseudocolor image, I would
convert the pseudocolor image to rgb and use the RGB gray merge
plugin. I guess that I don't understand what you mean by "or the
information of the pseudocolor gets lost as the image got flattened to
a single color" as well.

Post by Johannes Schindelin
But I guess I did not understand what you mean by "or the information of
the pseudocolor gets lost as the image got flattened to a single color".
Maybe you want to be a little more verbose about it, so I have a chance to
understand what you need?
Ciao,
Dscho

Alicia Piera
Lehrstuhl für Experimentalphysik:
Physik weicher Materie und Biophysik
Geschwister-Scholl-Platz 1
80539 München


Tel.: [phone]

Fax: [phone]

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