Yang Weijian, Yuste Rafael
Department of Electrical and Computer Engineering, University of California, Davis, CA, USA.
Neurotechnology Center, Department of Biological Sciences, Columbia University, New York, NY, USA.
Adv Exp Med Biol. 2021;1293:613-639. doi: 10.1007/978-981-15-8763-4_43.
A critical neuroscience challenge is the need to optically image and manipulate neural activity with high spatiotemporal resolution over large brain volumes. The last three decades have seen the development of calcium imaging to record activity from neuronal populations, as well as optochemistry and optogenetics to optically manipulate neural activity. These methods are typically implemented with wide-field or laser-scanning microscopes. While the former approach has a good temporal resolution, it generally lacks spatial resolution or specificity, particularly in scattering tissues such as the nervous system; meanwhile, the latter approach, particularly when combined with two-photon excitation, has high spatial resolution and specificity but poor temporal resolution. As a new technique, holographic microscopy combines the advantages of both approaches. By projecting a holographic pattern on the brain through a spatial light modulator, the activity of specific groups of neurons in 3D brain volumes can be imaged or stimulated with high spatiotemporal resolution. In a combination of other techniques such as fast scanning or temporal focusing, this high spatiotemporal resolution can be further improved. Holographic microscopy enables all-optical interrogating of neural activity in 3D, a critical tool to dissect the function of neural circuits.
神经科学面临的一项关键挑战是,需要在大的脑区范围内以高时空分辨率对神经活动进行光学成像和操纵。在过去三十年中,已经开发出钙成像技术来记录神经元群体的活动,以及光化学和光遗传学技术来光学操纵神经活动。这些方法通常通过宽场显微镜或激光扫描显微镜来实现。虽然前一种方法具有良好的时间分辨率,但通常缺乏空间分辨率或特异性,特别是在诸如神经系统等散射组织中;与此同时,后一种方法,特别是与双光子激发相结合时,具有高空间分辨率和特异性,但时间分辨率较差。作为一种新技术,全息显微镜结合了这两种方法的优点。通过空间光调制器在大脑上投射全息图案,可以以高时空分辨率对三维脑区中特定神经元群体的活动进行成像或刺激。与快速扫描或时间聚焦等其他技术相结合,这种高时空分辨率可以进一步提高。全息显微镜能够对三维神经活动进行全光学检测,是剖析神经回路功能的关键工具。