Journal
AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM
Volume 318, Issue 2, Pages E87-E101Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajpendo.00457.2019
Keywords
cell metabolism; diabetes; fluorescence microscopy; image analysis; live-cell imaging
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Funding
- Canadian Institutes of Health Research (CIHR) [MOP-119537]
- JDRF [2-2013-50]
- Canadian Foundation for Innovation [30214]
- Canada Graduate Scholarship from CIHR
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Live-cell imaging of mitochondrial function and dynamics can provide vital insights into both physiology and pathophysiology, including of metabolic diseases like type 2 diabetes. However, without super-resolution microscopy and commercial analysis software, it is challenging to accurately extract features from dense multilayered mitochondrial networks, such as those in insulin-secreting pancreatic beta-cells. Motivated by this, we developed a comprehensive pipeline and associated ImageJ plugin that enables 2D/3D quantification of mitochondrial network morphology and dynamics in mouse beta-cells and by extension other similarly challenging cell types. The approach is based on standard confocal microscopy and shareware, making it widely accessible. The pipeline was validated using mitochondrial photolabeling and unsupervised cluster analysis and is capable of morphological and functional analyses on a per-organelle basis, including in 4D (xyzt). Overall, this tool offers a powerful framework for multiplexed analysis of mitochondrial state/function and provides a valuable resource to accelerate mitochondrial research in health and disease.
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