31 October 2016 Label-free imaging of rat spinal cords based on multiphoton microscopy
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As an integral part of the central nervous system, the spinal cord is a communication cable between the body and the brain. It mainly contains neurons, glial cells, nerve fibers and fiber tracts. The recent development of the optical imaging technique allows high-resolution imaging of biological tissues with the great potential for non-invasively looking inside the body. In this work, we evaluate the imaging capacity of multiphoton microscopy (MPM) based on second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) for the cells and extracellular matrix in the spinal cord at molecular level. Rat spinal cord tissues were sectioned and imaged by MPM to demonstrate that MPM is able to show the microstructure including white matter, gray matter, ventral horns, dorsal horns, and axons based on the distinct intrinsic sources in each region of spinal cord. In the high-resolution and high-contrast MPM images, the cell profile can be clearly identified as dark shadows caused by nuclei and encircled by cytoplasm. The nerve fibers in white matter region emitted both SHG and TPEF signals. The multiphoton microscopic imaging technique proves to be a fast and effective tool for label-free imaging spinal cord tissues, based on endogenous signals in biological tissue. It has the potential to extend this optical technique to clinical study, where the rapid and damage-free imaging is needed.
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Chenxi Liao, Chenxi Liao, Zhenyu Wang, Zhenyu Wang, Linquan Zhou, Linquan Zhou, Xiaoqin Zhu, Xiaoqin Zhu, Wenge Liu, Wenge Liu, Jianxin Chen, Jianxin Chen, } "Label-free imaging of rat spinal cords based on multiphoton microscopy", Proc. SPIE 10024, Optics in Health Care and Biomedical Optics VII, 1002425 (31 October 2016); doi: 10.1117/12.2245519; https://doi.org/10.1117/12.2245519

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