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coloc2 plugin

Posted by Laure Twyffels on Jan 21, 2016; 10:33am
URL: http://imagej.273.s1.nabble.com/coloc2-plugin-tp5015422.html

Dear all,

 

I have a few questions about the output of the coloc2 plugin.

 

1)      How are the “Ch1 Max threshold” and “Ch2 Max threshold” calculated?
Do they correspond to Costes thresholding?  

2)      How are the Pearson’s “below threshold” and “above threshold” R
values calculated? Do they correspond to the values obtained for the pixels
in which at least one channel is below its “Max threshold” and for the
pixels in which both channels are above their “Max thresholds”,
respectively? Which use would you make of these values?

3)      Is Costes thresholding used for the Manders “threshold”
coefficients?

 

And finally, a more general question: I have analyzed only a few images
until now, but for all of them, and all my ROIs, I get a message saying that
“The absolute y-intercept of the auto threshold regression line is too
high”, and suggesting that I should use a ROI or do a background
subtraction. However, the values of the y-intercept do not seem to me
excessively high (e.g. -1, +3); what I consider as background has a low mean
value too, even inside ROIs. Also, I feel it strange to subtract a
background before asking the plugin to calculate a threshold that is used to
determine what “background” is. What is your opinion about that?

 

Thank you for your help !

 

Laure

 

Laure Twyffels

Center for Microscopy and Molecular Imaging (CMMI)

Université Libre de Bruxelles

 


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