Posted by
Theresa Swayne on
Feb 10, 2014; 9:29pm
URL: http://imagej.273.s1.nabble.com/To-make-a-image-from-several-images-tp5006451p5006478.html
Hello,
From this it sounds like you want to overlay channels in different colors.
Merging channels or changing the color (LUT = look-up table) does not change the data.
However, converting to RGB may change the data. (Intensity values from 0-4095 are compressed into a range from 0-255, depending on the display contrast settings.)
Therefore, if you are planning to convert a set of images to RGB for presentation, for each channel, set the display contrast (Image > Adjust > Brightness/Contrast) to the same Minimum and Maximum levels in all images before conversion. Each channel can have a different contrast setting, but all red channels in the figure should be the same as each other, for example.
I can provide more details offline if needed.
As many followers of this list can tell you, visual inspection has its place, but it's not the most convincing way to show colocalization.
See, for example,
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3074624/.
Hope this helps.
Theresa
On Feb 10, 2014, at 5:12 AM, JoeML <
[hidden email]> wrote:
> Thanks your for the help but I have some doubt yet. I want to create a image
> without modify here properties because i just want to observe if different
> marks co-localize. The way you say my image is not modified?
>
>>
>> If you want to merge the images as multiple channels:
>> 1. Use Image > Color > Channels Tool, click More >> Merge Channels… and select your images (don't worry that it says red/green/blue -- you will change it later) and check the box for Create Composite.
>> 2. Restore each image to grayscale: in Channels Tool, choose Color mode, check Channel 1, then More >> Grays. Do the same for each channel. Then More >> Convert to RGB to get a single image.
------------------------------------
Theresa Swayne, Ph.D.
Associate Research Scientist
Manager, Confocal and Specialized Microscopy Shared Resource
Herbert Irving Comprehensive Cancer Center, Columbia University
1130 Saint Nicholas Ave, 222A
New York, NY 10032
212-851-4613
[hidden email]
http://hiccc.columbia.edu/research/sharedresources/confocal--
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