Quantitative Analysis Of Mitochondrial Dynamics Illuminates Links Between Mitochondrial Dynamics And Cell Function, And Allows Single Organelle Localisation Microscopy

Mitochondrial structure is believed to be important to the normal function of cells. However, remarkably, in fully differentiated cells understanding of how mitochondrial structure controls cell function is preliminary. An ability to generate high resolution, high speed images of mitochondrial structure simultaneously with measurements of cell function is required. We present a suite of quantitative bio‐imaging tools developed for the measurement and analysis of mitochondrial dynamics from high speed fluorescence imaging of live cells, including the measurement and quantification of mitochondrial motion to sub‐diffraction limits, Single Organelle Localisation Microscopy, the use of transient depolarisation's of the mitochondrial membrane to segment physically and optically crowded clusters of mitochondria, and the co‐localization of mitochondria and Ca2+ signalling . We present the algorithmic basis of these tools and highlight their use in the context of changing mitochondrial dynamics in cellular proliferation and hypertension, with an emphasis on the generation of quantified measurements to yield insight into the different cellular states. These tools present a powerful combination of experimental procedures that give an unobstructed open view of new, previously hidden, mechanisms that control mitochondrial dynamics and cell function. Grant Funding Source : Wellcome Trust; British Heart Foundation; EPSRC