Flexible polygon-mirror based laser scanning microscope platform for multiphoton in-vivo imaging.

Area of Science:

• Biomedical Engineering
• Neuroscience
• Microscopy

Background:

• Commercial microscopy often uses fixed scanning speeds, limiting flexibility.
• Capturing fast biological events in vivo requires advanced imaging capabilities.
• Motion artifacts from physiological processes degrade image quality in small animal studies.

Purpose of the Study:

• To develop a high-performance, flexible multiphoton microscope system.
• To enable dynamic control of scanning speed and frame rates for in-vivo imaging.
• To integrate motion correction for enhanced image quality in live animal studies.

Main Methods:

• Constructed a polygon-mirror based multiphoton microscope with a dynamic line scanning system.
• Developed custom open-source software (PScan1.0) for digital control of scanning parameters.
• Implemented an image registration algorithm for real-time and post-processed motion correction.

Main Results:

• Achieved dynamic line scanning speeds from 2.7 kHz to 27 kHz and variable frame rates from 5 Hz to 50 Hz (512x512).
• Successfully captured calcium waves and transient peaks in single neurons at adjusted imaging speeds (40-160 Hz).
• Demonstrated effective motion correction, verified by improved blood flow rate quantification.

Conclusions:

• The developed system offers high performance and flexibility for in-vivo biological imaging.
• Dynamic scanning control allows optimization for signal gain or high-speed event capture.
• Integrated motion correction significantly improves the quality of real-time in-vivo cellular imaging.