Using MitER for 3D analysis of mitochondrial morphology and ER contacts

Therese Kichuk, Satyen Dhamankar, Saurabh Malani, William A. Hofstadter, Scott A. Wegner, Ileana M. Cristea, José L. Avalos

Research output: Contribution to journalArticlepeer-review

Abstract

We have developed an open-source workflow that allows for quantitative single-cell analysis of organelle morphology, distribution, and inter-organelle contacts with an emphasis on the analysis of mitochondria and mitochondria-endoplasmic reticulum (mito-ER) contact sites. As the importance of inter-organelle contacts becomes more widely recognized, there is a concomitant increase in demand for tools to analyze subcellular architecture. Here, we describe a workflow we call MitER (pronounced “mightier”), which allows for automated calculation of organelle morphology, distribution, and inter-organelle contacts from 3D renderings by employing the animation software Blender. We then use MitER to quantify the variations in the mito-ER networks of Saccharomyces cerevisiae, revealing significantly more mito-ER contacts within respiring cells compared to fermenting cells. We then demonstrate how this workflow can be applied to mammalian systems and used to monitor mitochondrial dynamics and inter-organelle contact in time-lapse studies.

Original languageEnglish (US)
Article number100692
JournalCell Reports Methods
Volume4
Issue number1
DOIs
StatePublished - Jan 22 2024

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Genetics
  • Radiology Nuclear Medicine and imaging
  • Computer Science Applications

Keywords

  • CP: Imaging
  • Saccharomyces cerevisea
  • image analysis
  • imaging
  • inter-organelle contact
  • mitochondrial-ER contact
  • organelle distribution
  • organelle morphology

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