High-Sensitivity Intrinsic Optical Signal Imaging Through Flexible, Low-Cost Adaptations of an Upright Microscope.
High-Sensitivity Intrinsic Optical Signal Imaging Through Flexible, Low-Cost Adaptations of an Upright Microscope.
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DOI：
10.1523/eneuro.0046-23.2023
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发表时间：
2023-07
期刊：
影响因子：
3.4
通讯作者：
Zeiger, William
中科院分区：
文献类型：
--
作者：
Vasquez, Brenda;Campos, Baruc;Cao, Ashley;Theint, Aye Theint;Zeiger, William
内在光信号成像（IOSI）是现代神经科学中的一项重要技术。30多年前，IOSI允许在整个皮层中宏观映射神经元活动。该技术已被用于研究感觉处理和经验依赖的可塑性，并经常被用作定位皮质区域的后续目标由其他成像或生理技术的一个连续的程序。尽管IOSI在神经科学中无处不在，但很少有商业上可用的交钥匙IOSI系统。因此，研究人员通常采用建立自己的成像系统。多年来，已经开发了作为专用钻机或并入现有显微镜平台的简化系统。在这里，我们提出了一套简单的适应，可以应用于任何标准的直立显微镜，使用现成的，廉价的，商业照明，光学和信号检测部分，使高灵敏度IOSI。利用这些适应性，我们能够很容易地绘制体感和视觉皮层的感觉诱发信号，包括小鼠的单须桶皮层活动图。我们表明，这些IOSI地图是高度可重复的动物，可用于研究可塑性机制的躯体感觉皮层。我们还提供开源应用程序来控制照明和分析原始数据以生成活动地图。我们预计，这些资源将是有用的神经科学研究人员希望增加IOSI能力，以现有的显微镜在实验室的预算。
Intrinsic optical signal imaging (IOSI) is a staple technique in modern neuroscience. Pioneered >30 years ago, IOSI allows macroscopic mapping of neuronal activity throughout the cortex. The technique has been used to study sensory processing and experience-dependent plasticity, and is often used as an adjunctive procedure to localize cortical areas for subsequent targeting by other imaging or physiology techniques. Despite the ubiquity of IOSI in neuroscience, there are few commercially available turn-key IOSI systems. As a result, investigators have typically resorted to building their own imaging systems. Over the years, simplified systems built either as dedicated rigs or incorporated into existing microscope platforms have been developed. Here we present a straightforward set of adaptations that can be applied to any standard upright microscope, using readily available, inexpensive, commercial parts for illumination, optics, and signal detection, that enables high-sensitivity IOSI. Using these adaptations, we are able to readily map sensory-evoked signals across the somatosensory and visual cortex, including single-whisker barrel cortical activity maps in mice. We show that these IOSI maps are highly reproducible across animals and can be used to study plasticity mechanisms in the somatosensory cortex. We also provide open-source applications to control illumination and analyze raw data to generate activity maps. We anticipate that these resources will be useful for neuroscience investigators looking to add IOSI capabilities to an existing microscope in the laboratory on a budget.