219 - Automated analysis of fluorescence micrographs demonstrates the importance of physiologically relevant O2 and glucose in live cell mitochondrial experiments

Abstract

Mitochondrial morphology, organization, and dynamics is intricately tied to the redox metabolism of animal cells. The physiological relevance of mitochondrial dynamics and how mitochondrial morphology relates to cell growth, stress, and metabolism has attracted much interest recently. Morphological investigation, addressing mitochondrial morphology and function in response to things such as oxidative stress and metabolic function, is often carried out by live cell fluorescence microscopy. For such investigations to be physiologically relevant, it is paramount that metabolic conditions reflect realistic ranges. Furthermore, as it becomes easier to generate large volumes of image data, unbiased and robust methods of analysing the morphology, organization, and dynamics of mitochondria for comparative purposes has become a necessity. Here, we introduce a refined version of our ImageJ plugin (MiNA 3.0) for analyzing mitochondrial morphology and organization in 2D and 3D, as well as through time. The plugin effectively captured the more fragmented morphology associated with excess glucose availability and elevated oxygen concentrations when compared with physiological conditions in common model cell lines. The work thus demonstrates the relevance of environmental control in live cell work and the benefit of semi-quantitative, automated analysis practices.