How to add a composite image stack as the additional channel to the a multi-channel Z stack?

I have 3-channel z stack images from confocal microscopy, and I want to check their co-localizations. So I need to have their composite images as the fourth channel.
It is quite easy to get the composite z stack by “split chennels/merge channels”, but then how do you attach it to the original z stack?

Hi @Ye_Jiahao,

you could split the channels of the image, generate new image stacks e.g. by multiplying channels using the Image Calculator ( https://imagej.nih.gov/ij/plugins/calculator-plus.html ) and finally merge the images as channels together again.

However, for colocalisation there are actually useful tools like


http://imagejdocu.tudor.lu/doku.php?id=plugin:analysis:jacop_2.0:just_another_colocalization_plugin:start

So why not using these tools?

This link may also be interesting for you:

Cheers,
Robert

Hi, Robert, Thanks a lot, especially for the info. on plugins for coloclization analysis. I will try them out!

If I am not mistaken, you are suggesting me an alternative way to merge multiple single-channel image into a composite image. But I actually stuck when I try to combine this composite image stack(i.e.256x256; c:1; z:30) with the original stacks (i.e. 256x256; c:3; z:30) into a new stack (then 256x256, c:4, z:30). It is very easy to do this with Leica LAS Lite, but I have not find a convenient with FIJI.

Cheers,
Jiahao

Just split your stack with three channels (using Image > Color > Split Channels… ) and combine these three individual images with another 4th image. I mean, in the menu Image > Color > Merge Channels, you can enter up to 7 images. They all need to be of size 256x256; c:1; z:30 in your case :wink:

Cheers,
Robert