High-speed two-photon microscopy with adaptive line-excitation

Yunyang Li; Shu Guo; Ben Mattison; Junjie Hu; Kwun Nok Mimi Man; Weijian Yang

doi:10.1364/OPTICA.529930

High-speed two-photon microscopy with adaptive line-excitation

Optica. 2024 Aug 15;11(8):1138-1145. doi: 10.1364/OPTICA.529930. eCollection 2024 Aug 20.

Authors

Yunyang Li¹, Shu Guo¹, Ben Mattison^{2

3}, Junjie Hu¹, Kwun Nok Mimi Man^{4

5}, Weijian Yang^{1

3}

Affiliations

¹ Department of Electrical and Computer Engineering, University of California, Davis, California 95616, USA.
² Department of Biomedical Engineering, University of California, Davis, California 95616, USA.
³ Biomedical Engineering Graduate Group, University of California, Davis, California 95616, USA.
⁴ Department of Biochemistry and Molecular Medicine, University of California, Davis, California 95616, USA.
⁵ Current address: Max Planck Florida Institute for Neuroscience, Jupiter, Florida 33458, USA.

Abstract

We present a two-photon fluorescence microscope designed for high-speed imaging of neural activity at cellular resolution. Our microscope uses an adaptive sampling scheme with line illumination. Instead of building images pixel by pixel via scanning a diffraction-limited spot across the sample, our scheme only illuminates the regions of interest (i.e., neuronal cell bodies) and samples a large area of them in a single measurement. Such a scheme significantly increases the imaging speed and reduces the overall laser power on the brain tissue. Using this approach, we performed high-speed imaging of the neuronal activity in mouse cortex in vivo. Our method provides a sampling strategy in laser-scanning two-photon microscopy and will be powerful for high-throughput imaging of neural activity.

Associated data

figshare/10.6084/m9.figshare.26139925