用于记录自由活动动物神经活动的微型多光子显微镜

Miniature Multiphoton Microscopes for Recording Neural Activity in Freely Moving Animals

作者信息

Ozbay Baris N., Futia Gregory L., Ma Ming, McCullough Connor, Young Michael D., Restrepo Diego, Gibson Emily A.

机构信息

Department of Bioengineering, University of Colorado Denver, Aurora, CO, USA

Department of Cell and Developmental Biology, University of Colorado Denver, Aurora, CO, USA

DOI:10.1007/978-1-0716-2764-8_7

PMID:38285825

Abstract

Miniaturized head-mounted microscopes for in vivo recording of neural activity have gained much recognition within the past decade of neuroscience research. In combination with fluorescent reporters, these miniature microscopes allow researchers to record the neural activity that underlies behavior, cognition, and perception in freely moving animals. Single-photon miniature microscopes are convenient for widefield recording but lack the increased penetration depth and optical sectioning capabilities of multiphoton imaging. Here we discuss the current state of head-mounted multiphoton miniature microscopes and introduce a miniature head-mounted two-photon fiber-coupled microscope (2P-FCM) for neuronal imaging with active axial focusing enabled using a miniature electrowetting lens. The 2P-FCM enables three-dimensional two-photon optical recording of structure and activity at multiple focal planes in a freely moving mouse. Detailed methods are provided in this chapter on the 2P-FCM design, operation, and software for data analysis.

摘要

在过去十年的神经科学研究中，用于体内记录神经活动的小型头戴式显微镜已获得广泛认可。结合荧光报告基因，这些微型显微镜使研究人员能够记录自由活动动物行为、认知和感知背后的神经活动。单光子微型显微镜便于进行宽场记录，但缺乏多光子成像所具有的增加穿透深度和光学切片能力。在这里，我们讨论头戴式多光子微型显微镜的当前状态，并介绍一种微型头戴式双光子光纤耦合显微镜（2P-FCM），用于通过使用微型电润湿透镜实现主动轴向聚焦的神经元成像。2P-FCM能够在自由活动的小鼠体内对多个焦平面的结构和活动进行三维双光子光学记录。本章提供了有关2P-FCM设计、操作和数据分析软件的详细方法。