目的利用三维建模的方法对耳蜗内毛细胞带状突触进行计数分析,解决由于内毛细胞带状突触数量少、位置深在造成的计数困难,为内毛细胞带状突触的可塑性研究提供有效可靠的计数方法。方法取小鼠耳蜗基底膜,使用免疫荧光双标记的方法对突触...目的利用三维建模的方法对耳蜗内毛细胞带状突触进行计数分析,解决由于内毛细胞带状突触数量少、位置深在造成的计数困难,为内毛细胞带状突触的可塑性研究提供有效可靠的计数方法。方法取小鼠耳蜗基底膜,使用免疫荧光双标记的方法对突触前 RIBEYE 和突触后膜 GluR2&3 进行标记,激光共聚焦显微镜进行光学连续切片,每个荧光色对代表一个突触的存在。使用 3ds max 进行三维建模,对内毛细胞带状突触进行计数。结果耳蜗基底膜内毛细胞的带状突触显示清晰,每个内毛细胞的带状突触数量为(16.10±1.03)个。结论利用免疫荧光双标,激光共聚焦显微镜光学连续切片,3ds max 三维建模所得的内毛细胞带状突触数量准确,方法简单可行,是一种对内毛细胞带状突触进行计数的可靠方法。展开更多
In mammals,the ribbon synapses of cochlear inner hair cells are a synaptic structure of the first sensory nerve in the pathway of acoustical signal transmission to the acoustic center,and it is directly involved in vo...In mammals,the ribbon synapses of cochlear inner hair cells are a synaptic structure of the first sensory nerve in the pathway of acoustical signal transmission to the acoustic center,and it is directly involved in voice coding and neurotransmitter release. It is difficult to quantitatively analyze the ribbon synaptic number only using an electron microscope,because the ribbon synaptic number is relatively limited and their location is deep. In this study,the specific presynaptic structure-RIBEYE,and non-specific postsynaptic structure-GluR 2 & 3 in C57BL/6J mouse basilar membrane samples were treated by immunofluorescent labeling. Serial section was performed on the samples using a laser scanning confocal microscope,and then the serial sections were used to build three-dimensional models using 3DS MAX software. Each fluorescein color pair indicates one synapse,so the number of ribbon synapses of inner hair cells is obtained. The spatial distribution and the number of ribbon synapses of cochlear inner hair cells were clearly shown in this experiment,and the mean number of ribbon synapses per inner hair cell was 16.10±1.03. Our results have demonstrated the number of ribbon synapses is accurately calculated by double immunofluorescent labeling to presynaptic and postsynaptic structures,serial sections obtained using a laser scanning confocal microscope,and three-dimensional modeling obtained using 3DS MAX software. The method above is feasible and has important significance for further exploring the mechanism of sensorineural deafness.展开更多
The optical recording of three-dimensional(3-D)reconstruction of CA1 pyramidal cells wasderived from the studies on the CA1 region of the hippocampus in adult male Wistar rats.The recordingwas produced by the Confocal...The optical recording of three-dimensional(3-D)reconstruction of CA1 pyramidal cells wasderived from the studies on the CA1 region of the hippocampus in adult male Wistar rats.The recordingwas produced by the Confocal Laser Scan Microscope(LSM-10).The attemption was to outline themorphological neural network of CA1 pyramidal cells organization,following the trail of axo-dendritic connec-tions in 3-D spatial distributions among neurons.The fractal structure of neurons with their dendritic andaxonal trees using fractal algorithm was noticed,and 2—18 simulated cells were obtained using PC-486 comput-er.The simulational cells are similar in morphology to the natural CA1 hippocampal pyramidal cells.There-fore,the exploitation of an advanced neurohistological research technique combining optical recording of theLSM-10 and computer simulation of fractal structure can provide the quantitative fractal structural basis forchaosic dynamics of brain.展开更多
文摘目的利用三维建模的方法对耳蜗内毛细胞带状突触进行计数分析,解决由于内毛细胞带状突触数量少、位置深在造成的计数困难,为内毛细胞带状突触的可塑性研究提供有效可靠的计数方法。方法取小鼠耳蜗基底膜,使用免疫荧光双标记的方法对突触前 RIBEYE 和突触后膜 GluR2&3 进行标记,激光共聚焦显微镜进行光学连续切片,每个荧光色对代表一个突触的存在。使用 3ds max 进行三维建模,对内毛细胞带状突触进行计数。结果耳蜗基底膜内毛细胞的带状突触显示清晰,每个内毛细胞的带状突触数量为(16.10±1.03)个。结论利用免疫荧光双标,激光共聚焦显微镜光学连续切片,3ds max 三维建模所得的内毛细胞带状突触数量准确,方法简单可行,是一种对内毛细胞带状突触进行计数的可靠方法。
文摘In mammals,the ribbon synapses of cochlear inner hair cells are a synaptic structure of the first sensory nerve in the pathway of acoustical signal transmission to the acoustic center,and it is directly involved in voice coding and neurotransmitter release. It is difficult to quantitatively analyze the ribbon synaptic number only using an electron microscope,because the ribbon synaptic number is relatively limited and their location is deep. In this study,the specific presynaptic structure-RIBEYE,and non-specific postsynaptic structure-GluR 2 & 3 in C57BL/6J mouse basilar membrane samples were treated by immunofluorescent labeling. Serial section was performed on the samples using a laser scanning confocal microscope,and then the serial sections were used to build three-dimensional models using 3DS MAX software. Each fluorescein color pair indicates one synapse,so the number of ribbon synapses of inner hair cells is obtained. The spatial distribution and the number of ribbon synapses of cochlear inner hair cells were clearly shown in this experiment,and the mean number of ribbon synapses per inner hair cell was 16.10±1.03. Our results have demonstrated the number of ribbon synapses is accurately calculated by double immunofluorescent labeling to presynaptic and postsynaptic structures,serial sections obtained using a laser scanning confocal microscope,and three-dimensional modeling obtained using 3DS MAX software. The method above is feasible and has important significance for further exploring the mechanism of sensorineural deafness.
基金the National Natural Science Foundation of ChinaLaboratory of Visual Information Processing,Institute of Biophysics,Chinese Academy of Sciences
文摘The optical recording of three-dimensional(3-D)reconstruction of CA1 pyramidal cells wasderived from the studies on the CA1 region of the hippocampus in adult male Wistar rats.The recordingwas produced by the Confocal Laser Scan Microscope(LSM-10).The attemption was to outline themorphological neural network of CA1 pyramidal cells organization,following the trail of axo-dendritic connec-tions in 3-D spatial distributions among neurons.The fractal structure of neurons with their dendritic andaxonal trees using fractal algorithm was noticed,and 2—18 simulated cells were obtained using PC-486 comput-er.The simulational cells are similar in morphology to the natural CA1 hippocampal pyramidal cells.There-fore,the exploitation of an advanced neurohistological research technique combining optical recording of theLSM-10 and computer simulation of fractal structure can provide the quantitative fractal structural basis forchaosic dynamics of brain.
