Real-time imaging of cell cleaning processes in mice using advanced two-photon microscopy

A computationally enhanced imaging framework enables high-resolution analysis of autophagy dynamics in vivo.

• The imaging system integrates two-photon microscopy with an optimized tissue stabilization method to reduce motion artifacts.
• Real-time imaging captures LC3-positive structures in the liver and skeletal muscle under various stress conditions.
• Image quality is improved through a trained denoising model and enhanced Super-Resolution Radial Fluctuations.
• Autophagic flux is assessed using chloroquine to differentiate between increased autophagosome formation and impaired degradation.
• This framework addresses significant technical challenges in intravital imaging, allowing for a detailed analysis of autophagy.

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