Faculty Collaboration Database

Methods for imaging mammalian mitochondrial morphology: A prospective on MitoGraph. Anal Biochem 2018 Jul 01;552:81-99

Date

03/06/2018

Pubmed ID

29505779

Pubmed Central ID

PMC6322684

DOI

10.1016/j.ab.2018.02.022

Scopus ID

2-s2.0-85048014256 (requires institutional sign-in at Scopus site)   53 Citations

Abstract

Mitochondria are found in a variety of shapes, from small round punctate structures to a highly interconnected web. This morphological diversity is important for function, but complicates quantification. Consequently, early quantification efforts relied on various qualitative descriptors that understandably reduce the complexity of the network leading to challenges in consistency across the field. Recent application of state-of-the-art computational tools have resulted in more quantitative approaches. This prospective highlights the implementation of MitoGraph, an open-source image analysis platform for measuring mitochondrial morphology initially optimized for use with Saccharomyces cerevisiae. Here Mitograph was assessed on five different mammalian cells types, all of which were accurately segmented by MitoGraph analysis. MitoGraph also successfully differentiated between distinct mitochondrial morphologies that ranged from entirely fragmented to hyper-elongated. General recommendations are also provided for confocal imaging of labeled mitochondria (using mito-YFP, MitoTracker dyes and immunostaining parameters). Widespread adoption of MitoGraph will help achieve a long-sought goal of consistent and reproducible quantification of mitochondrial morphology.

Author List

Harwig MC, Viana MP, Egner JM, Harwig JJ, Widlansky ME, Rafelski SM, Hill RB

Authors

Megan C. Harwig Research Scientist II in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin
Michael E. Widlansky MD Associate Director, Professor in the Medicine department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Bacterial Proteins
Biomarkers
Cell Line
Humans
Luminescent Proteins
Mitochondria
Transfection