Re: Color Merge Multiple Fluorescence Images
Posted by
John Oreopoulos on
Jan 21, 2010; 3:54pm
URL: http://imagej.273.s1.nabble.com/Color-Merge-Multiple-Fluorescence-Images-tp3689609p3689611.html
See color submenu here:
http://rsbweb.nih.gov/ij/docs/menus/image.html#colorRGB Merge
Merges 1-4 greyscale images or stacks into an RGB image or stack.
Select *None* to keep a channel empty (filled with 0). Check "Create
Composite" to convert 2-4 grayscale images or stacks into a composite
image or hyperstack. Check "Keep Source Images" if you wish to keep
the originals.
Adjust the brightness and contrast of your channel images first (and
make sure you say that you have done this in your figure captions or
your methods sections! - See Rossner, M., and K. M. Yamada. 2004.
What's in a picture? The temptation of image manipulation. J. Cell
Biol. 166:11-15). Then run the RGB merge command. If you have many
image sets to do this on, you can automate by writing a simple macro
in ImageJ
John Oreopoulos
On 21-Jan-10, at 3:58 AM, Christian Breukers wrote:
> Dear all,
>
>
>
> I'm working on a fluorescence microscope that has four fluorescence
> channels (DAPI, FITC, PE, APC).
>
>
>
> Since we are measuring antibody conjugated fluorescent cells, we
> have a
> large variation in intensity and background.
>
>
>
> What would be the correct way to have a color merge of four
> fluorescence
> microscope images / channels?
>
> What pre processing steps are necessary to modify the images such
> that a
> nice clear color overlay is created?
>
>
>
> Kind regards,
>
> Christian Breukers
>
> ---------------------------------------------------
> University of Twente
>
> Faculty of Sciences and Technology
>
> MIRA Institute for Biomedical Technology and Technical Medicine
>
> Department of Medical Cell BioPhysics
> ---------------------------------------------------
>
>