3D super-resolution deep-tissue imaging in living mice.

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Stimulated emission depletion (STED) microscopy provides 3D super-resolution imaging of dynamic nanoscale structures in living cells.
• Deep-tissue imaging with 3D-STED is limited by optical aberrations and light scattering.

Purpose of the Study:

• To develop and demonstrate a STED system capable of overcoming deep-tissue imaging challenges.
• To achieve aberration-corrected 3D super-resolution imaging deep within biological tissues.

Main Methods:

• Integration of two-photon excitation with adaptive optics.
• Utilization of red-emitting organic dyes and a long-working-distance water-immersion objective lens.
• Implementation of a novel STED microscopy system.

Main Results:

• Demonstrated aberration-corrected 3D super-resolution imaging at 164 µm depth in fixed mouse brain tissue.
• Achieved 76 µm depth imaging in the brain of a living mouse with high resolution.
• Successfully overcame optical aberrations and light scattering for deep-tissue STED.

Conclusions:

• The developed STED system significantly advances the capability for deep-tissue super-resolution microscopy.
• This technology offers unprecedented insights into the organization of nanoscale structures in vivo.
• The system holds promise for future biological and biomedical research requiring deep-tissue imaging.