High-speed miniaturized two-photon microscopy with elliptical beam excitation

Ben Mattison; Shing-Jiuan Liu; Feng Tian; Weijian Yang

doi:10.1117/12.3001090

13 March 2024 High-speed miniaturized two-photon microscopy with elliptical beam excitation

Ben Mattison, Shing-Jiuan Liu, Feng Tian, Weijian Yang

Proceedings Volume PC12828, Neural Imaging and Sensing 2024; PC1282806 (2024) https://doi.org/10.1117/12.3001090
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Two-photon microscopy traditionally suffers from slow frame rates due to the necessity of raster scanning the laser excitation spot over the field of view (FOV). We have developed a miniaturized elliptical beam shaper (MEBS) which is compatible with the state-of-the-art open-source two-photon miniscopes for imaging neural activity in freely moving mice. The MEBS enables an elliptical two-photon excitation pattern with a long axis of 5-10 µm, which greatly reduces the number of scanning rows required while maintaining cellular resolution in imaging. As a result, the overall frame rate is increased compared to the diffraction-limited-point scanning method. We demonstrate this high-speed miniaturized two-photon microscope in fluorescence imaging and show a significant frame rate improvement while maintaining cellular resolution across a 400 µm diameter FOV.

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Citation Download Citation

Ben Mattison, Shing-Jiuan Liu, Feng Tian, and Weijian Yang "High-speed miniaturized two-photon microscopy with elliptical beam excitation", Proc. SPIE PC12828, Neural Imaging and Sensing 2024, PC1282806 (13 March 2024); https://doi.org/10.1117/12.3001090

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KEYWORDS

Miniaturization

Two photon excitation microscopy

Microscopes

Functional imaging

Image resolution

Objectives

Optical fibers

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