Research progress of miniature head-mounted single photon fluorescence microscopic imaging technique
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摘要:
微型头戴式单光子荧光显微成像技术是近些年出现的用于神经科学研究的一种突破性方法，可以对自由移动活体动物的神经活动进行实时成像，提供了一种前所未有的方式来访问神经信号，增强了对大脑如何工作的理解。在脑科学研究需求的推动下，目前已经出现了许多种类型的微型头戴式单光子荧光显微镜，如高分辨率成像、无线记录、三维成像、双区域成像和双色成像等。为了更加全面地了解和认识这种新兴的光学神经成像技术，本文按成像视场进行分类，对目前报道的不同类型微型头戴式单光子荧光显微镜所具有的特点进行了介绍，重点讨论了其所采用的光学系统方案和光学性能参数，分析对比了不同方案的优缺点，以及未来的改进方向，以便为脑科学研究人员的实际应用提供参考。
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关键词:
- 微型单光子荧光显微镜 /
- 神经信号 /
- 脑科学 /
- 光学系统
Abstract:Miniature head-mounted single-photon fluorescence microscopy is a breakthrough approach for neuroscience research that has emerged in recent years. It can image the neural activity of freely moving vivo animals in real time, providing an unprecedented way to access neural signals and rapidly enhancing the understanding of how the brain works. Driven by the needs of brain science research, there have been many types of miniature head-mounted single-photon fluorescence microscopes, such as high-resolution imaging, wireless recording, 3D imaging, two-region imaging and two-color imaging. In order to have a more comprehensive understanding of this new optical neuroimaging technology, we classify its technologies according to the imaging field of view, introduce the characteristics of different types of micro-head-mounted single-photon fluorescence microscopes reported so far, and focus on the optical system scheme and optical performance parameters used. The advantages and disadvantages of different schemes are analyzed and compared and the future direction of development is described to provide reference for the practical application of brain science researchers.
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图 1 具有基本成像功能的系统。（a）Ghosh等人的集成显微镜的横截面图[10]；（b）MiniScope V3的分解图; （c）戴着微型显微镜的小鼠示意图[14]；（d）小鼠大脑中神经元活动的荧光图像[14]
Figure 1. A system with a basic imaging function. (a) Cross sectional view of integrated microscope proposed by Ghosh et al; (b) exploded view of the MiniScope V3; (c) a schematic of a mouse wearing a miniature microscope; (d) fluorescent images of neural activity in a mouse brain
图 3 具有三维成像功能的系统。（a）MiniLFM的横截面图[24]；（b）Miniscope3D的横截面图[27];（c）Bagramyan等人的显微镜横截面图[28]；（d）SIMscope3D的横截面图[29]
Figure 3. A system with 3D imaging functionality. (a) Cross sectional view of MiniLFM; (b) cross sectional view of Miniscope3D; (c) microscope cross section by Bagramyan et al; (d) cross sectional view of SIMscope3D
表 1 具有基本成像功能的微型荧光显微镜的光学系统和光学性能参数
Table 1. Optical system and optical performance parameters of the miniature fluorescence microscope with basic imaging functionality
系统参数 Ghosh 等人 MiniScope V3 miniscope CHEndoscope Bagramyan 等人 物镜 梯度折射率透镜 梯度折射率透镜 非球面透镜 梯度折射率透镜 梯度折射率透镜 管镜 双胶合透镜 双胶合透镜 双胶合透镜 双胶合透镜 平凸透镜 视场 600 μm×800 μm 750 μm×450 μm 1100 μm×1100 μm ~500 μm ~105 μm 分辨率 2.5 μm 1.0 μm/pix 单细胞分辨率 单细胞分辨率 1.0 μm 图像传感器 MT9V021
(5.6 μm/pix)MT9V032
(6.0 μm/pix)MT9V022
(6.0 μm/pix)MT9P031
(2.2 μm/pix)OV7251
(3.0 μm/pix)成像速度 36 Hz 60 Hz 10 Hz 20 Hz 50 Hz 
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表 2 具有无线功能的微型荧光显微镜的光学系统和光学性能参数
Table 2. Optical system and optical performance parameters of a miniature fluorescence microscope with wireless function
系统参数 FinchScope Wire-free MiniScope miniscope wScope 物镜 梯度折射率透镜 梯度折射率透镜 非球面透镜 梯度折射率透镜 管镜 双胶合透镜 双胶合透镜 双胶合透镜 双胶合透镜 视场 800 μm×600 μm — 500 μm×500 μm 700 μm×450 μm 分辨率 单细胞分辨率 1 μm/pix 单细胞分辨率 1.8 μm 图像传感器 OV7960(6.00 μm/pix) EV76C454(5.80 μm/pix) MT9V022(6.00 μm/pix) OV7690 (1.75 μm/pix) 成像速度 30 Hz 10 Hz 10 Hz 25 Hz
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表 3 具有三维成像功能的微型荧光显微镜的光学系统和光学性能参数
Table 3. Optical system and optical performance parameters of the miniature fluorescence microscope with 3D imaging functionality
系统参数 MiniLFM Miniscope3D Bagramyan等人 OMKAR 等人 物镜 梯度折射率透镜 梯度折射率透镜 梯度折射率透镜 两片双胶合透镜 管镜 双胶合透镜 相位掩模板 平凸透镜 双胶合透镜 视场 700 μm×600 μm×360 μm 900 μm×700 μm×390 μm 横向 150 μm
轴向 98 μm横向207 μm
轴向220 μm三维成像元件 微透镜阵列 相位掩模板 可调谐液晶透镜 电湿润透镜 横向分辨率 6.0 μm 2.8 μm 1.4 μm 1.0 μm/pix 轴向分辨率 30.0 μm 15.0 μm 15.0 μm 18.0 μm 图像传感器 MT9V032 (6.0 μm/pix) MT9V032 (6.0 μm/pix) OV7251 (3.0 μm/pix) MT9P031 (2.2 μm/pix) 成像速度 16 Hz 40 Hz 50 Hz —
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表 4 具有双区域成像功能的微型荧光显微镜的光学系统和光学性能参数
Table 4. Optical system and optical performance parameters of a miniature fluorescence microscope with dual region imaging functionality
系统参数 Gonzalez 等人 NINscope 物镜 梯度折射率透镜 梯度折射率透镜 管镜 双胶合透镜 平凸透镜 视场 600 μm×479 μm 786 μm×502 μm 分辨率 0.83 μm/pix 单细胞分辨率 图像传感器 OV7690 (6 μm/pix) PYTHON480 (4.8 μm/pix) 成像速度 — 30 Hz
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表 5 具有双色成像功能的微型荧光显微镜的光学系统和光学性能参数
Table 5. Optical system and optical performance parameters of a miniature fluorescence microscope with two-color imaging functionality
系统参数 MiniScope V4 DCFIMM-SBI DCFIMM-DBI 物镜 两片双胶合透镜 两片双胶合透镜 双胶合透镜 管镜 双胶合透镜 双胶合透镜 双胶合透镜 视场 ~1.00 mm2 1.10 mm×1.10 mm 0.77 mm×0.77 mm 分辨率 单细胞分辨率 3.47 μm 3.47 μm 图像传感器 PYTHON480
(4.8 μm/pix)EV76C454
(5.8 μm/pix)EV76C454
(5.8 μm/pix)成像速度 120 Hz 20 Hz 20 Hz
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表 6 小视场微型单光子荧光显微镜的光学系统组成和光学性能参数
Table 6. Optical system composition and optical performance parameters of miniature single photon fluorescence microscope with a small field
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表 7 大视场微型荧光显微镜的光学系统和光学性能参数
Table 7. Optical system and optical performance parameters of a large field miniature fluorescence microscope
cScope CM2 mScope 物镜 多片球面透镜 微透镜阵列 双凸透镜 管镜 多片球面透镜 视场 7.8 mm×4.0 mm 7.3 mm×8.1 mm×
2.5 mm8.0 mm×10.0 mm 横向分辨率 14.0 μm 7 μm 39.4~55.7 μm 图像传感器 MT9V032
(6 µm/pix)MT9P031
(2.2 µm/pix)MT9V032
(6 µm/pix)成像速度 60Hz — —
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