Colocalization

Analysis in Fluorescence Micrographs: Verification of a More Accurate Calculation of Pearson’s Correlation Coefficient.

Colocalization Analysis in Fluorescence Micrographs

This recently published paper in Microscopy and Microanalysis is co-authored by PerkinElmer Application Specialist Dr. Andrew Barlow, Senior Software Developer Alasdair MacLeod, and colleagues at the University of Ghent, Belgium:

Barlow A L, MacLeod A, Noppen S, Sanderson J and Guérin C J (2010). Colocalization Analysis in Fluorescence Micrographs: Verification of a More Accurate Calculation of Pearson’s Correlation Coefficient. Microscopy and Microanalysis doi: 10.1017/S143192761009389X.

Abstract: One of the most routine uses of fluorescence microscopy is colocalization, i.e. the demonstration of a relationship between pairs of biological molecules. Frequently this is presented simplistically by the use of overlays of red and green images, with areas of yellow indicating colocalization of the molecules. Colocalization data is rarely quantified, and even when it is can still be misleading. Our results from both theoretical and biological datasets demonstrate that the mathematical models currently used produce results which overestimate the positive correlation of molecules and fail to demonstrate negative correlations. We have found that the calculation of a thresholded Pearson’s correlation coefficient using only intensity values over a determined threshold in both channels produces numerical values which more accurately describe both synthetic datasets and biological examples. Its use will bring clarity and accuracy to colocalization studies using fluorescent microscopy. 

This paper has been accepted for publication and is shown below in its revised form, subsequent to editorial input by Cambridge University Press, in Microscopy and Microanalysis, published by Cambridge University Press © 2010 Microscopy Society of America.

View the full text of this article – revised version (PDF)
View the figures  – revised  version  (PDF)

View the final abstract and full text (requires a subscription) on the Cambridge Journals homepage