Fast wavefront shaping for two-photon brain imaging with multipatch correction.

Area of Science:

• Biomedical Optics
• Microscopy
• Neuroscience

Background:

• Nonlinear fluorescence microscopy enables deep in vivo imaging of cellular structures.
• Adaptive wavefront correction enhances imaging depth but is limited by a small field of view.
• Existing methods struggle with aberrations and scattering in thick biological tissues.

Purpose of the Study:

• To develop a novel acousto-optic light modulation technique for simultaneous wavefront correction and fluorescence imaging.
• To overcome the field-of-view limitations of current adaptive optics in scattering media.
• To achieve high-resolution in vivo imaging at increased depths and speeds.

Main Methods:

• Introduced an acousto-optic light modulation technique for fluorescence imaging.
• Implemented simultaneous, on-the-fly wavefront correction synchronized with pixel scanning.
• Employed adaptive optimization to learn biaxial wavefront corrections at multiple image locations.
• Utilized raster scanning with position-dependent correction switching.

Main Results:

• Demonstrated simultaneous wavefront correction and fluorescence imaging at pixel scan speed (40 kHz).
• Successfully imaged neurons in vivo through a thinned skull, correcting for aberrations and scattering.
• Achieved multi-patch correction, extending the effective field of view for aberration correction.

Conclusions:

• The acousto-optic technique significantly enhances the depth and speed of in vivo nonlinear fluorescence microscopy.
• This method overcomes field-of-view limitations of adaptive optics, enabling deeper and wider imaging in scattering tissues.
• The technology shows promise for advanced neuroscience research requiring high-resolution imaging of neural activity in vivo.