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Related Experiment Videos

Protocol for evaluating mitochondrial morphology changes in response to CCCP-induced stress through open-source image

Sudeshna Nag1, Kaitlin Szederkenyi2, Christopher M Yip2

  • 1Department of Biochemistry, University of Toronto, MaRS Centre West Tower, 661 University Avenue, M5G 1M1 Toronto, Canada.

STAR Protocols
|December 1, 2023
PubMed
Summary

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This summary is machine-generated.

Quantifying mitochondrial morphology is complex. This protocol simplifies it by using an aspect ratio to categorize mitochondria into subclasses, aiding cellular health assessment under stress.

Area of Science:

  • Cell Biology
  • Mitochondrial Biology
  • Cellular Physiology

Background:

  • Mitochondrial morphology reflects cellular health and function.
  • Quantifying and categorizing mitochondrial subclasses is challenging.
  • Standardized protocols are needed for reliable assessment.

Purpose of the Study:

  • To present a protocol for quantifying mitochondrial morphology.
  • To enable categorization of mitochondria into subclasses.
  • To assess mitochondrial morphology under carbonyl cyanide m-chlorophenyl hydrazone (CCMPH) stress.

Main Methods:

  • Protocol for cell preparation for immunofluorescence microscopy.
  • Staining techniques for mitochondria.
  • Morphology quantification using aspect ratio calculation.
Keywords:
Cell BiologyCell cultureMetabolismMicroscopyMolecular/Chemical Probes

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Main Results:

  • A reliable method for mitochondrial morphology quantification is described.
  • The protocol allows categorization into two distinct mitochondrial subclasses.
  • Mitochondrial morphology changes under CCMPH stress can be assessed.

Conclusions:

  • The presented protocol simplifies mitochondrial morphology quantification.
  • The aspect ratio method effectively categorizes mitochondria.
  • This protocol aids in evaluating cellular health and response to stress.