High frame-rate multichannel beam-scanning microscopy based on Lissajous trajectories

Area of Science:

• Optical Imaging
• Microscopy
• Biophysics

Background:

• High-speed optical imaging is crucial for capturing dynamic biological processes.
• Existing beam-scanning methods often face limitations in frame rate or resolution.
• Simultaneous acquisition of diverse data channels presents a significant challenge.

Purpose of the Study:

• To develop a simple beam-scanning optical design for kHz frame-rate imaging.
• To enable simultaneous acquisition of multiple data channels.
• To demonstrate the viability of Lissajous trajectory imaging for sensitive detection.

Main Methods:

• Utilized two fast-scan resonant mirrors to create a Lissajous trajectory for beam scanning.
• Employed data segmentation and model-based image reconstruction (MBIR) with 3D in-painting algorithms.
• Integrated control and data acquisition electronics with modified beam-scanning hardware.

Main Results:

• Achieved effective frame rates significantly exceeding the Lissajous trajectory repeat time.
• Demonstrated simultaneous acquisition of high-frame rate (low resolution) and low-frame rate (high resolution) images.
• Preliminary studies confirmed viability using laser transmittance imaging and second harmonic generation microscopy.

Conclusions:

• Lissajous trajectory imaging offers a viable approach for high-speed, multi-channel optical imaging.
• The technique allows for flexible trade-offs between image resolution and frame rate.
• It shows promise for detecting subtle changes and transient fluctuations in optical signals.