Non-telecentric two-photon microscopy for 3D random access mesoscale imaging.

Area of Science:

• Neuroscience
• Optical Imaging
• Microscopy

Background:

• Conventional two-photon microscopy has limitations in field-of-view size and excitation volume shape.
• These constraints restrict the scope of biological questions and obtainable information in neuronal activity monitoring.

Purpose of the Study:

• To present a low-cost, flexible solution to overcome the limitations of conventional two-photon microscopy.
• To expand the three-dimensional field-of-view and enable advanced imaging capabilities.

Main Methods:

• Utilized a non-telecentric optical design.
• Incorporated rapid laser-focus control using an electrically tunable lens.
• Developed a system easily implemented and reversible within hours.

Main Results:

• Achieved a several-fold expansion of the three-dimensional field-of-view.
• Enabled near-simultaneous imaging of remote 3D regions.
• Demonstrated the ability to bend imaging planes to follow biological curvatures.
• Successfully imaged neuronal activity in zebrafish, mice, and fruit flies.

Conclusions:

• The presented system offers a flexible and cost-effective advancement for two-photon microscopy.
• The expanded imaging capabilities facilitate a broader range of neuroscience research.
• The adaptable design serves as a valuable platform for future developments in optical monitoring.