Quantifying mitochondrial content in living cells

Matheus Palhares Viana; Swee Lim; Susanne M Rafelski

doi:10.1016/bs.mcb.2014.10.003

Quantifying mitochondrial content in living cells

Methods Cell Biol. 2015:125:77-93. doi: 10.1016/bs.mcb.2014.10.003. Epub 2015 Jan 8.

Authors

Matheus Palhares Viana¹, Swee Lim², Susanne M Rafelski³

Affiliations

¹ Departments of Developmental and Cell Biology, University of California, Irvine, CA, USA.
² Biomedical Engineering, University of California, Irvine, CA, USA.
³ Departments of Developmental and Cell Biology, University of California, Irvine, CA, USA; Biomedical Engineering, University of California, Irvine, CA, USA; Center for Complex Biological Systems, University of California, Irvine, CA, USA.

PMID: 25640425
DOI: 10.1016/bs.mcb.2014.10.003

Abstract

We describe a novel version of MitoGraph, our fully automated image processing method and software, dedicated to calculating the volume of 3D intracellular structures and organelles in live cells. MitoGraph is optimized and validated for quantifying the volume of tubular mitochondrial networks in budding yeast. We therefore include the experimental protocol, microscopy conditions, and software parameters focusing on mitochondria in budding yeast. However, MitoGraph can also be applied to mitochondria in other cell types and possibly other intracellular structures. We begin with our protocol and then include substantial discussion of the validation, requirements, and limits of MitoGraph to aid a wide range of potential users in applying MitoGraph to their data and troubleshooting any potential problems that arise. MitoGraph is freely available at the Web site http://rafelski.com/susanne/MitoGraph.

Keywords: Budding yeast; Microscopy; MitoGraph; Mitochondria; Networks; Skeletonization; Software; Topology; Tubules; Volume.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Line, Tumor
Cell Survival
Humans
Image Processing, Computer-Assisted / methods*
Microscopy, Confocal
Mitochondria / metabolism*
Mitochondrial Dynamics
Mutation
Saccharomyces cerevisiae / cytology*
Saccharomyces cerevisiae / metabolism*
Software