To make a image from several images

[JoeML]

Hello everyone

I have a doubt about to create a only image from two or three images. I tried doing a stack but when i try to "make composite"  it doesnt work or modifies the original grayscale. Anybody know some way to make a only image from several images (2 or 3) and keep those images in grayscale.

Thanks you very much

[Krs5]

What about Image > Stacks > Images to Stack?

Kees

-----Original Message-----
From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of JoeML
Sent: 07 February 2014 17:26
To: [hidden email]
Subject: To make a image from several images

Hello everyone

I have a doubt about to create a only image from two or three images. I tried doing a stack but when i try to "make composite"  it doesnt work or modifies the original grayscale. Anybody know some way to make a only image from several images (2 or 3) and keep those images in grayscale.

Thanks you very much



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[Theresa Swayne]

Hello,

I'm not sure if I understand what you want, but here are some possibilities:

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.

If you want a projection (e.g. with each pixel replaced by the maximum, minimum, or average of that position in all images), make a stack of the images and then Image > Stacks > Z Project.

If you want a montage (multiple panels), make a stack and Image > Stacks > Make Montage.

Hope one of these ideas is helpful.

Theresa

On Feb 7, 2014, at 12:25 PM, JoeML <[hidden email]> wrote:

>
> I have a doubt about to create a only image from two or three images. I
> tried doing a stack but when i try to "make composite"  it doesnt work or
> modifies the original grayscale. Anybody know some way to make a only image
> from several images (2 or 3) and keep those images in grayscale.
>
>

------------------------------------
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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[JoeML]

Hi everybody

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?

Thanks you very much for all your help

[Theresa Swayne]

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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