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Analyzing Mitochondrial Morphology Through Simulation Supervised Learning
Published on: March 3, 2023
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
Summary
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.
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.
