Ten-kilohertz two-photon microscopy imaging of single-cell dendritic activity and hemodynamics in vivo

Ruijie Li; Sibo Wang; Jing Lyu; Ke Chen; Xiaxin Sun; Junjie Huang; Pei Sun; Susu Liang; Min Li; Mengke Yang; Hongbang Liu; Shaoqun Zeng; Xiaowei Chen; Longhui Li; Hongbo Jia; Zhenqiao Zhou

doi:10.1117/1.NPh.10.2.025006

3 May 2023 Ten-kilohertz two-photon microscopy imaging of single-cell dendritic activity and hemodynamics in vivo

Ruijie Li, Sibo Wang, Jing Lyu, Ke Chen, Xiaxin Sun, Junjie Huang, Pei Sun, Susu Liang, Min Li, Mengke Yang, Hongbang Liu, Shaoqun Zeng, Xiaowei Chen, Longhui Li, Hongbo Jia, Zhenqiao Zhou

Author Affiliations +

Neurophotonics, Vol. 10, Issue 2, 025006 (May 2023). https://doi.org/10.1117/1.NPh.10.2.025006

Abstract

Significance

The studying of rapid neuronal signaling across large spatial scales in intact, living brains requires both high temporal resolution and versatility of the measurement device.

Aim

We introduce a high-speed two-photon microscope based on a custom-built acousto-optic deflector (AOD). This microscope has a maximum line scan frequency of 400 kHz and a maximum frame rate of 10,000 frames per second (fps) at 250 × 40 pixels. For stepwise magnification from population view to subcellular view with high spatial and temporal resolution, we combined the AOD with resonance-galvo (RS) scanning.

Approach

With this combinatorial device that supports both large-view navigation and small-view high-speed imaging, we measured dendritic calcium propagation velocity and the velocity of single red blood cells (RBCs).

Results

We measured dendritic calcium propagation velocity (80 / 62.5 − 116.7 μm / ms) in OGB-1-labeled single cortical neurons in mice in vivo. To benchmark the spatial precision and detection sensitivity of measurement in vivo, we also visualized the trajectories of single RBCs and found that their movement speed follows Poiseuille’s law of laminar flow.

Conclusions

This proof-of-concept methodological development shows that the combination of AOD and RS scanning two-photon microscopy provides both versatility and precision for quantitative analysis of single neuronal activities and hemodynamics in vivo.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Ruijie Li, Sibo Wang, Jing Lyu, Ke Chen, Xiaxin Sun, Junjie Huang, Pei Sun, Susu Liang, Min Li, Mengke Yang, Hongbang Liu, Shaoqun Zeng, Xiaowei Chen, Longhui Li, Hongbo Jia, and Zhenqiao Zhou "Ten-kilohertz two-photon microscopy imaging of single-cell dendritic activity and hemodynamics in vivo," Neurophotonics 10(2), 025006 (3 May 2023). https://doi.org/10.1117/1.NPh.10.2.025006

Received: 3 January 2023; Accepted: 3 April 2023; Published: 3 May 2023

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KEYWORDS

Calcium

Hemodynamics

Two photon imaging

Neurons

In vivo imaging

Red blood cells

Two photon excitation microscopy

Significance

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