Deciphering the fine morphology and precise location of neurons and neural circuits are crucial to enhance our understanding of brain function and diseases. Traditionally, we have to map brain images to coarse axial-sampling planar reference atlases to orient neural structures. However, this means might fail to orient neural projections at single-cell resolution due to position errors resulting from individual differences at the cellular level. Here, we present a high-throughput imaging method that can automatically obtain the fine morphologies and precise locations of both neurons and circuits, employing wide-field large-volume tomography to acquire three-dimensional images of thick tissue and implementing real-time soma counterstaining to obtain cytoarchitectonic landmarks during the imaging process. The reconstruction and orientation of brain-wide neural circuits at single-neuron resolution can be accomplished for the same mouse brain without additional counterstains or image registration. Using our method, mouse brain imaging datasets of multiple type-specific neurons and circuits were successfully acquired, demonstrating the versatility. The results show that the simultaneous acquisition of labeled neural structures and cytoarchitecture reference at single-neuron resolution in the same brain greatly facilitates precise tracing of long-range projections and accurate locating of nuclei. Our method provides a novel and effective tool for application in studies on genetic dissection, brain function and the pathology of the nervous system.
Qingming Luo, Hui Gong, Jing Yuan, Xiangning Li, Anan Li, and Tonghui Xu, "High-throughput dual-color precision imaging for brain-wide mapping of the connectome with cytoarchitectonic landmarks at the cellular level (Conference Presentation)," Proc. SPIE 10051, Neural Imaging and Sensing, 100510D (Presented at SPIE BiOS: January 30, 2017; Published: 19 April 2017); https://doi.org/10.1117/12.2251624.5371358589001.
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