Fig. 7
From: Pitfalls in methods to study colocalization of nanoparticles in mouse macrophage lysosomes
NP colocalization with lysosomes between fixed-and live-cell imaging. A Experimental workflow. Representative images of a single cell exposed to two different NP types, B SiO2-RhoB NP (green) and C SiO2-BDP FL NP (green). Lysosomes were stained with LAMP-2 antibody (red). The corresponding histograms of fluorescence intensities of each channel are shown in Additional file 1: Fig. S5B. The graphs depict the main parameters (Pearson’s correlation coefficient—PCC, Manders’ correlation coefficients—M1 and M2) to estimate the colocalization between D SiO2-RhoB NP and lysosomes (LysoTracker Green) and E SiO2-BDP FL NP and lysosomes (LysoTracker Red) in live cells (white box) and fixed cells stained with LAMP-2 (grey box). The whiskers are the standard deviations. The analysis was done in individual cells for each experiment (n = 8–13 cells). All the images were analysed using ImageJ with the JACoP plugin comparing the correlation coefficients. Statistical analysis was performed using unpaired t-test in GraphPad Prism software. ***p < 0.001, ns: not significant