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Pupil plane actuated remote focusing for rapid focal depth control
Optics Express
|September 10, 2020
Summary
We developed a new remote focusing method for laser scanning microscopy that avoids actuator damage and achieves sub-millisecond response times. This technique enables faster in vivo imaging of neural activity in mouse brains.
Area of Science:
- Optics and Photonics
- Neuroscience
- Biomedical Engineering
Background:
- Laser scanning microscopy is crucial for imaging and material processing, with existing systems excelling at 2D scanning.
- Rapid control of focal depth is essential for advanced applications, but current remote focusing methods risk actuator damage.
- Laser-induced damage to actuators near the focal point limits the speed and reliability of remote focusing techniques.
Purpose of the Study:
- To introduce a novel remote focusing method that overcomes laser-induced damage limitations.
- To achieve rapid, sub-millisecond focal depth control in laser scanning systems.
- To demonstrate the efficacy of this method in advanced microscopy for in vivo neural imaging.
Main Methods:
- Developed a pupil plane actuated remote focusing technique.
- Implemented a dual-plane two-photon laser scanning fluorescence microscope.
- Utilized the new method for in vivo calcium transient recording in mouse neocortex.
Main Results:
- Achieved sub-millisecond response times for focal depth control.
- Successfully avoided laser-induced damage to the focusing actuator.
- Enabled high-speed, in vivo recording of neuronal calcium transients in live mouse brains.
Conclusions:
- Pupil plane actuated remote focusing offers a damage-free solution for rapid focal control.
- This method significantly enhances the capabilities of laser scanning fluorescence microscopy.
- The technique is suitable for high-resolution, in vivo imaging of dynamic biological processes.
