Opto2P-FCM: A MEMS based miniature two-photon microscope with two-photon patterned optogenetic stimulation.

Area of Science:

• Neuroscience
• Biomedical Engineering
• Microscopy

Background:

• Multiphoton microscopy and optogenetics are crucial for understanding neural circuits in live animals.
• Current limitations hinder simultaneous neural imaging and patterned optogenetic stimulation in freely moving subjects.
• Previous techniques struggled with spatial precision in freely moving animal models.

Purpose of the Study:

• To develop a compact, head-mountable microscope for high-resolution two-photon imaging.
• To integrate spatially patterned two-photon optogenetic stimulation capabilities.
• To enable cell-specific neural activity studies in freely moving animals.

Main Methods:

• Designed a miniature microscope incorporating a MEMS scanner for two-photon imaging.
• Integrated a secondary beam path for patterned two-photon optogenetic stimulation.
• Developed a lightweight, head-attachable system for freely moving subjects.

Main Results:

• Demonstrated high-resolution two-photon fluorescence imaging using a MEMS scanner.
• Achieved cell-specific optogenetic stimulation with patterned two-photon excitation.
• Successfully imaged neural activity in freely moving mice with the miniature microscope.

Conclusions:

• The developed miniature microscope overcomes previous limitations in freely moving animal studies.
• This technology facilitates cell-specific investigation of neural mechanisms underlying behavior.
• Enables unprecedented insights into neural dynamics in naturalistic behaviors.