Exploring and Understanding Colocalization Metrics
Colocalization analysis of microscopy images is a very sought-after measurement that is actually quite nuanced and seldom implemented correctly.
In this video we are going to go over some of the most commonly used colocalization metrics and explain the strengths and weaknesses of each, so you can choose which one to use and how.
Presentation slides available in zenodo: https://zenodo.org/records/17081523
Chapters:
0:00 Intro
0:44 What is Colocalization?
7:34 Considerations for Image acquisition
13:10 Correlation and Co-occurrence
15:55 Colocalization metrics - Area Overlap
19:39 Colocalization metrics - Pearson's Correlation Coefficient (PCC)
34:55 Colocalization metrics - Spearman's Rank Correlation Coefficient (SRCC)
37:03 Colocalization metrics - Manders' Overlap Coefficient (MOC)
39:39 Colocalization metrics - Manders' Correlation Coefficients (MCCs)
45:10 Thresholding method - Coste's Automatic Threshold
47:55 Statystical test - Coste's Randomization
51:14 Colocalization metrics - Rank-weighed Colocalization Coefficient
51:54 Which colocalization metric should I use?
52:49 Take-home messages
References:
Aaron JS et al., 2018, Image co-localization - co-occurrence versus correlation, DOI: 10.1242/jcs.211847
Dunn KW et al., 2011, A practical guide to evaluating colocalization in biological microscopy, DOI: 10.1152/ajpcell.00462.2010
López S et al., 2024, On the pixel selection criterion for the calculation of the Pearson’s correlation coefficient in fluorescence microscopy, DOI: doi.org/10.1111/jmi.13273
Adler J & Parmryd I, 2010, Quantifying colocalization by correlation: the Pearson correlation coefficient is superior to the Mander's overlap coefficient, DOI: 10.1002/cyto.a.20896
Adler J & Parmryd I, 2021, Quantifying colocalization: The case for discarding the Manders overlap coefficient, DOI: 10.1002/cyto.a.24336
Bolte S & Cordelieres FP, 2006, A guided tour into subcellular colocalization analysis in light microscopy, DOI: 10.1111/j.1365-2818.2006.01706.x
Costes SV et al., 2004, Automatic and Quantitative Measurement of Protein-Protein Colocalization in Live Cells, DOI: 10.1529/biophysj.103.038422
Manders EMM et al., 1993, Measurement of co-localization of objects in dual-colour confocal images, DOI: 10.1111/j.1365-2818.1993.tb03313.x
Singan VR et al., 2011, Dual channel rank-based intensity weighting for quantitative co-localization of microscopy images, DOI: 10.1186/1471-2105-12-407