Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins

Area of Science:

• Cell Biology
• Microscopy Techniques
• Physiology

Background:

• Live imaging of subcellular structures is crucial for understanding cellular dynamics.
• Existing in vivo imaging methods have limitations in spatial resolution and accessibility.
• Salivary glands offer unique advantages for microscopy due to accessibility and ease of manipulation.

Purpose of the Study:

• To describe a novel confocal intravital microscopy protocol for imaging subcellular structures in live rodents.
• To demonstrate the application of this technique in studying secretory granule exocytosis and apical plasma membrane dynamics in mouse salivary glands.
• To establish a robust system for investigating biological processes at a molecular level in vivo.

Main Methods:

• Utilized confocal intravital microscopy for high-resolution live imaging.
• Employed live mouse salivary glands as a model organ, stabilizing them to minimize motion artifacts.
• Used transgenic mice expressing cytosolic GFP and membrane-targeted tandem-Tomato for subcellular labeling.

Main Results:

• Successfully imaged subcellular structures and tracked the kinetics of secretory granule exocytosis.
• Visualized the dynamics of the apical plasma membrane during stimulated exocytosis.
• Demonstrated the feasibility of extending the protocol to other mouse models and labeling strategies.

Conclusions:

• Confocal intravital microscopy provides a powerful tool for high-resolution in vivo imaging of subcellular dynamics.
• The described protocol enables detailed investigation of cellular processes like exocytosis in accessible organs.
• This technique has broad applicability for visualizing diverse subcellular components in various biological contexts.