Hi Steela,

I think your questions are answered in this paper

https://imagejdocu.tudor.lu/_media/plugin/analysis/jacop_2.0/just_another_colocalization_plugin/jacop_ijconf2008.pdf . It describes the thresholding options and how they are implemented in JACOP.

You should also read the original paper if you havenâ€™t already.

Kind regards,

Jacqui

From: ImageJ Interest Group <

[hidden email]> On Behalf Of Steela Hong

Sent: Tuesday, February 2, 2021 12:07 AM

To:

[hidden email]
Subject: Help with JACoP plugin

Hi,

I am using the JACoP plugin in ImageJ for analysing colocalisation of two

proteins.

The output looks like this:

Image A: Image 1.lsm - C=1

Image B: Image 1.lsm - C=2

Pearson's Coefficient:

r=0.522

Manders' Coefficients (original):

M1=0.67 (fraction of A overlapping B)

M2=0.598 (fraction of B overlapping A)

Manders' Coefficients (using threshold value of 2 for imgA and 11 for imgB):

M1=0.565 (fraction of A overlapping B)

M2=0.541 (fraction of B overlapping A)

Costes' automatic threshold set to 2 for imgA & 5 for imgB

Pearson's Coefficient:

r=0.413 (0.0 below thresholds)

M1=0.862 & M2=0.943

--------------------------------------------------------------------------------------------------

After the analysis, I get three sets of Mander's coefficient values.

My question is what is the thresholding method used here for calculating

Mander's coefficient with threshold. How does ImageJ determine the

threshold intensity for each channel?

And second, how is this thresholding method different from Coste's

automatic thresholding?

Thank you so much.

Steela Hong

PhD Candidate

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