Extended-resolution imaging of the interaction of lipid droplets and mitochondria

An imaging informatics pipeline enabling the high-resolution spatio-temporal analysis of the subcellular organization of lipid droplet-mitochondria associations in living COS-7 cells is presented. The settings of a popular image deconvolution tool for improvement of the quality of spatial images, the limitations of the image segmentation process, as well as the diffraction pattern of the subcellular compartments significantly affect the results of morphometric analyses. In this context, most lipid droplets showed temporary contacts to mitochondrial membranes rather than stable associations, however, the LD-mitochondria association was increased upon oleic acid treatment and upon overexpression of PLIN5 (Plin5, Perilipin 5).141