摘要
To visualize the structure and organization of the brain is a fundamental requirement in the research of neuroscience. Here, combining with two-photon excitation fluorescence microscopy and transgenetic mouse GAD67,we demonstrate a custom-built second harmonic generation（SHG） microscope to discriminate brain layers and sub regions in the cerebellum and brain stem slices with cellular resolution. In particular, the cell densities of neurons in different brain layers are extracted due to the cell soma appearing as dark shadow on an SHG image.Further, the axon initial segments of the Purkinje cell are easily recognized without labeling, which would be useful for guiding micropipettes for electrophysiology.
To visualize the structure and organization of the brain is a fundamental requirement in the research of neuroscience. Here, combining with two-photon excitation fluorescence microscopy and transgenetic mouse GAD67,we demonstrate a custom-built second harmonic generation（SHG） microscope to discriminate brain layers and sub regions in the cerebellum and brain stem slices with cellular resolution. In particular, the cell densities of neurons in different brain layers are extracted due to the cell soma appearing as dark shadow on an SHG image.Further, the axon initial segments of the Purkinje cell are easily recognized without labeling, which would be useful for guiding micropipettes for electrophysiology.
作者
Xiuli Liu
Daozhu Hua
Ling Fu
Shaoqun Zeng
刘秀丽;华道柱;付玲;曾绍群(Britton Chance Center for Biomedical Photonics,School of Engineering Sciences,Wuhan National Laboratory for Optoelectronlcs--Huazhong University of Science and Technology,Wuhan 430074,China;MoE Key Laboratory for Biomedical Photonics,Department of Biomedical Engineering,Huazhong University of Science and Technology,Wuhan 430074,China)
基金
supported by the National Key Research and Development Program of China(No.2016YFA0201403)
the National Natural Science Foundation of China(No.61522502)
the Science Fund for Creative Research Group of China(No.61421064)
