Sensorless adaptive optics implementation in widefield optical sectioning microscopy inside in vivo Drosophila brain

Area of Science:

• Biomedical Optics
• Microscopy
• Neuroscience

Background:

• Aberrations in fluorescence microscopy limit imaging depth and resolution.
• Adaptive optics (AO) can correct these aberrations but often requires complex setups.
• In vivo imaging of complex biological tissues like the Drosophila brain presents significant optical challenges.

Purpose of the Study:

• To implement a sensorless adaptive optics loop for widefield fluorescence microscopy.
• To compensate for sample-induced aberrations in both excitation and emission pathways.
• To enable fast optical sectioning within living Drosophila brains.

Main Methods:

• Developed a sensorless adaptive optics (AO) system integrated into a widefield fluorescence microscope.
• Utilized AO to correct aberrations introduced by the biological sample.
• Applied structured illumination microscopy (SIM) in conjunction with AO.

Main Results:

• Demonstrated effective compensation for aberrations in living Drosophila brains.
• Achieved fast optical sectioning capabilities at depths up to 50 μm.
• Quantified significant improvements in optical sectioning using AO-SIM.

Conclusions:

• Sensorless AO is a viable method for aberration correction in widefield fluorescence microscopy.
• This technique enables high-resolution imaging deep within scattering biological tissues.
• The developed system significantly enhances optical sectioning for in vivo neuroscience research.