Levet, F. et al. Nat. Commun. 10, 2379 (2019).

Single-molecule localization microscopy (SMLM) is becoming increasingly useful for quantitative analysis of cellular structures. Multicolor localization microscopy allows researchers to determine the relative positions of multiple structures, but parameters such as the degree of colocalization can be challenging to quantify, especially in 3D. Levet et al. have developed Coloc-Tesseler for analyzing colocalization in 2D and 3D SMLM data. Coloc-Tesseler begins by generating Voronoї diagrams for each color channel for image segmentation, which then enables standard colocalization metrics such as Manders and Pearson’s coefficients to be calculated. Coloc-Tesseler’s performance was validated on synthetic and experimental data, and it is likely to be a powerful tool for analyzing multicolor localization microscopy data. The software is implemented in a user-friendly GUI.