Integrated approaches for multiscale mitochondrial structure and function analysis.

Area of Science:

• Cell Biology
• Biophysics
• Microscopy

Background:

• Mitochondria are vital organelles responsible for ATP production, calcium homeostasis, and apoptosis.
• Visualizing mitochondrial dynamics and ultrastructure requires advanced imaging technologies across various scales.

Purpose of the Study:

• To review diverse mitochondrial imaging modalities, including their principles, capabilities, and limitations.
• To highlight recent advances in fluorescent probes, correlative techniques, and computational analysis for mitochondrial imaging.

Main Methods:

• Conventional fluorescence microscopy (wide-field, confocal, light-sheet) for live-cell dynamics.
• Super-resolution microscopy (SIM, STED, PALM, STORM, expansion microscopy) for sub-mitochondrial structures.
• Proximity-based assays (FRET, split-fluorescent proteins) and electron microscopy (EM) for interactions and ultrastructure.

Main Results:

• Diverse imaging techniques offer complementary spatial and temporal resolutions for mitochondrial studies.
• Novel probes, correlative methods, and computational tools enhance the utility of mitochondrial imaging.
• Super-resolution and EM provide insights into fine structures like cristae and inter-organelle contact sites.

Conclusions:

• Mitochondrial imaging has significantly advanced our comprehension of cellular physiology and pathology.
• The integration of various imaging modalities and computational approaches is key to future discoveries.
• Understanding the principles and applications of these techniques is crucial for biological research.