Quantitative Determination of Spatial Protein-protein Proximity in Fluorescence Confocal Microscopy

TL;DR

This paper presents a method to quantify spatial protein-protein proximity in fluorescence microscopy images by decomposing correlation functions into fast and slow components, improving reliability in colocalization analysis.

Contribution

It introduces a novel approach to separate specific protein clustering from background effects, enhancing accuracy in colocalization measurements.

Findings

Fast component correlates with specific protein clusters

Method improves reliability of proximity indices

Applicable to fluorescence confocal microscopy images

Abstract

To quantify spatial protein-protein proximity (colocalization) in fluorescence microscopic images, cross-correlation and autocorrelation functions were decomposed into fast and slowly decaying components. The fast component results from clusters of proteins specifically labeled and the slow one from background/image heterogeneity. We show that the calculation of the protein-protein proximity index and the correlation coefficient are more reliably determined by extracting the fast-decaying component.