Object based co-localization data interpretation



Hi All,

I am doing object based co-localization analysis using Jacop plugin. I have some difficulty in interpreting the results.
for example I got this results:

Colocalization based on distance between centres of mass
Threshold for Image A=63; Image B=49
Particles size between 0 & 1228800
Image A: 1404 centre(s) colocalizing out of 3179
Image B: 1559 centre(s) colocalizing out of 5122

if you calculate the percent of coloclaiztion fro image A it is 44.1 % while for image B is 30.4%. why are not the same ? and what percents represent a high and good co-localization versus low and poor colocaliztion ?

Thanks in advance,


Hi Banan and welcome to the forum!

The reason you have different numbers is that you have a different number of particles per image.
For example if you have 100 particles in image A and 1000 in image B, the maximum number of particles colocalization for A will be 100%, because that’s all there is in image A. This also means that the maximum colocalization in Image B will be 10%.

As to what represents a high, mid and low colocalization, welcome to the gray area of this technique! :wink:
You would need to provide us with some example images (in raw format) perhaps shared via a file sharing platform. In the case of object based colocalization, it depends both on the contents of the images and of the segmentation that you have applied.

In cases of colocalization in our platform, the first question is “What are you hoping to answer, biologically speaking, with a colocalization analysis?”



Hi Oburri,
Thanks for your helpful reply.
I am testing a drug conjugate that has red florescence (captured by RFP filter) and I stained the lysosomes with Lyso Tracker green (captured by GFP). I want to know the percent of co localization of my drug in these lysosomes ?
attached are example of my images

Thanks in advance,