RoundMi: A quantitative method to analyze mitochondrial morphology in mitotic cells

Area of Science:

• Cell Biology
• Biophysics
• Computational Biology

Background:

• Mitochondrial morphology reflects cellular health and disease.
• Quantitative analysis in mitotic cells is difficult due to their 3D structure.
• Current volumetric methods are resource-intensive and slow.

Purpose of the Study:

• To develop a streamlined workflow for analyzing mitochondrial morphology in mitotic cells.
• To enable rapid, quantitative assessment using single focal plane imaging.
• To overcome limitations of existing high-throughput methods.

Main Methods:

• Developed RoundMi, a workflow integrating Nellie plugin (Napari) and MitoSkel.
• Utilized single focal plane imaging guided by DNA and mitochondrial staining.
• Validated in mouse embryonic fibroblasts (MEFs) and HeLa cells.

Main Results:

• RoundMi accurately detects mitochondrial morphology differences between wild-type and DRP1-deficient cells.
• Single-plane analysis reliably approximates volumetric data without artifacts.
• Significantly reduced computational demand and acquisition time compared to volumetric methods.

Conclusions:

• RoundMi provides a versatile, high-throughput solution for mitochondrial morphology analysis in dividing cells.
• The method is compatible with live-cell imaging and applicable across cell types.
• Simplifies complex analyses, enhancing accessibility for researchers.