Understanding Mitochondrial and Lysosomal Dynamics by Fluorescent Microscopy.

Area of Science:

• Cell Biology
• Microscopy
• Bioimaging

Background:

• Live cell imaging visualizes dynamic cellular structures like mitochondria and lysosomes.
• Mitochondrial dynamics (fission/fusion) regulate organelle size and morphology.
• Lysosome size and distribution can indicate cellular function.

Purpose of the Study:

• To describe live cell confocal imaging of mitochondria and lysosomes.
• To present 2D and 3D analysis methods for organelle morphology.
• To utilize the Mitochondria Analyzer plugin for ImageJ/Fiji.

Main Methods:

• Live cell confocal microscopy was performed using MitoTracker and LysoTracker dyes.
• Organelle morphology and mitochondrial network connectivity were analyzed.
• The freely available Mitochondria Analyzer plugin for ImageJ/Fiji was employed for quantification.

Main Results:

• Successful live cell imaging of mitochondria and lysosomes was achieved.
• Quantitative analysis of mitochondrial morphology and lysosomal morphology was performed.
• The Mitochondria Analyzer plugin facilitated detailed 2D and 3D assessments.

Conclusions:

• Live cell imaging combined with the Mitochondria Analyzer plugin offers a robust approach for studying organelle dynamics.
• This method allows for detailed morphological and network analysis of mitochondria and lysosomes.
• The described protocol is valuable for investigating cellular processes involving these organelles.