Integration of Single-Photon Miniature Fluorescence Microscopy and Electrophysiological Recording Methods for in vivo Studying Hippocampal Neuronal Activity

Pub Date : 2024-08-29 DOI:10.1134/s0022093024040264

A. I. Erofeev, E. K. Vinokurov, I. E. Antifeev, O. L. Vlasova, I. B. Bezprozvanny

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Abstract

The miniature single-photon fluorescent microscope (miniscope) enables the visualization of calcium activity in vivo in freely moving laboratory animals, providing the capability to track cellular activity during the investigation of memory formation, learning, sleep, and social interactions. However, the use of calcium sensors for in vivo imaging is limited by their relatively slow (millisecond-scale) kinetics, which complicates the recording of high-frequency spike activity. The integration of methods from single-photon miniature fluorescent microscopy with electrophysiological recording, which possesses microsecond resolution, represents a potential solution to this issue. Such a combination of techniques allows for the simultaneous recording of optical and electrophysiological activity in a single animal in vivo. In this study, a flexible polyimide microelectrode was developed and integrated with the gradient lens of the miniscope. The in vivo tests conducted in this research confirmed that the microelectrode combined with the gradient lens facilitates simultaneous single-photon calcium imaging and local field potential recording in the hippocampus of an adult mouse.

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整合单光子微型荧光显微镜和电生理记录方法，在体内研究海马神经元活动

摘要微型单光子荧光显微镜（miniscope）能够观察自由活动的实验动物体内的钙离子活动，为研究记忆形成、学习、睡眠和社会交往过程中的细胞活动提供了能力。然而，使用钙传感器进行体内成像受到其相对较慢（毫秒级）动力学的限制，这使得记录高频尖峰活动变得复杂。将单光子微型荧光显微镜的方法与具有微秒分辨率的电生理记录相结合，是解决这一问题的潜在方法。将这两种技术相结合，可以同时记录单个动物体内的光学和电生理活动。本研究开发了一种柔性聚酰亚胺微电极，并将其与微型显微镜的梯度透镜集成在一起。本研究进行的体内测试证实，微电极与梯度透镜的结合有助于在成年小鼠的海马中同时进行单光子钙成像和局部场电位记录。

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