目的旨在建立脑组织样本神经血管单元中内皮细胞线粒体快速可视化分割的方法和应用。方法取假手术组纹状体和缺血再灌注组纹状体半暗区组织,采用锇-硫代碳酰肼-锇制样技术(Osmium-thiocarbohydrazide-osmium,OTO)包埋前块染色技术,结合...目的旨在建立脑组织样本神经血管单元中内皮细胞线粒体快速可视化分割的方法和应用。方法取假手术组纹状体和缺血再灌注组纹状体半暗区组织,采用锇-硫代碳酰肼-锇制样技术(Osmium-thiocarbohydrazide-osmium,OTO)包埋前块染色技术,结合聚焦离子束扫描电子显微镜(Focused ion beam scanning electron microscope,FIB/SEM)和人工智能可视化图像分析软件(Artificial intelligence-visualization-image analysis,AIVIA)机器学习图像分割技术,对神经血管单元中内皮细胞线粒体进行大尺度成像和快速分割分析。结果本研究成功建立基于OTO-FIB/SEM-AIVIA(OFA)的脑组织块样本线粒体快速分割流程,实现微米级神经血管单元中特定细胞类型线粒体纳米级分辨率的快速可视化;该流程可以提供脑组织样本线粒体数量和形态变化的准确信息;同时,训练出的模型可以拓展应用于相同图像采集参数的不同样本数据,提高图像分割速度和数据处理效率;基于这一方法,本研究发现相较于假手术组,脑缺血再灌注组纹状体半暗区内神经血管单元完整线粒体的平均体积变小且形态变圆;本研究还进一步提取并三维重建了内皮细胞中的线粒体并发现其形态和数量的改变。结论OFA流程可以快速获取神经血管单元或其中某个特定细胞类型,提供内皮细胞完整线粒体数量和体积的准确信息,为功能研究提供更准确全面的结构学证据。展开更多
A method for in situ preparation of fluorescent gold nanoclusters(Au NCs) with bovine serum albumin/montmorillonite composite powder(Au NC-BSA/MMT) was developed, and the products were used to detect latent fingermark...A method for in situ preparation of fluorescent gold nanoclusters(Au NCs) with bovine serum albumin/montmorillonite composite powder(Au NC-BSA/MMT) was developed, and the products were used to detect latent fingermarks. In this work, Au NCs were "grown" both inside and on the surface of BSA/MMT clay using one-step reduction of HAu Cl4 by BSA. The as-prepared Au NC-BSA/MMT nanocomposites emit intensive red fluorescence under the excitation of UV-visible light and show stable chemical features and low toxicity. The obtained fluorescent powders were characterized by UV-visible absorption spectroscopy,fluorescence spectroscopy, infrared spectroscopy, transmission electron microscopy/high-resolution transmission electron microscopy, scanning electron microscopy, X-ray photoelectron spectroscopy and X-ray diffraction to depict their sizes, structural information and optical features. Given their environmentally friendly preparation, simple operation, low cost, efficient UVvisible radiation-dependent photoluminescence and good affinity with finger residues, the in situ synthesized Au NC-BSA/MMT nanocomposite powders were used as an alternative fluorescent developing reagent for developing latent fingermarks deposited on various object surfaces(such as glass, aluminum foil, painted metal, plastic products and weighing papers) for individual identification. As results, the developed fingermarks with clear patterns and satisfactory level-2(minutiae points) and level-3(sweat pores) ridge details were obtained. Notably, treated prints could be excited by red light and emitted near infrared fluorescence, which was beneficial to avoid background interference and reduce the damage caused by UV light. With the advantages of the simple preparation process and good enhancement performance for latent fingermarks, the proposed method might be used in the preparation of various fluorescent probes for detecting trace evidence in forensic sciences.展开更多
文摘目的旨在建立脑组织样本神经血管单元中内皮细胞线粒体快速可视化分割的方法和应用。方法取假手术组纹状体和缺血再灌注组纹状体半暗区组织,采用锇-硫代碳酰肼-锇制样技术(Osmium-thiocarbohydrazide-osmium,OTO)包埋前块染色技术,结合聚焦离子束扫描电子显微镜(Focused ion beam scanning electron microscope,FIB/SEM)和人工智能可视化图像分析软件(Artificial intelligence-visualization-image analysis,AIVIA)机器学习图像分割技术,对神经血管单元中内皮细胞线粒体进行大尺度成像和快速分割分析。结果本研究成功建立基于OTO-FIB/SEM-AIVIA(OFA)的脑组织块样本线粒体快速分割流程,实现微米级神经血管单元中特定细胞类型线粒体纳米级分辨率的快速可视化;该流程可以提供脑组织样本线粒体数量和形态变化的准确信息;同时,训练出的模型可以拓展应用于相同图像采集参数的不同样本数据,提高图像分割速度和数据处理效率;基于这一方法,本研究发现相较于假手术组,脑缺血再灌注组纹状体半暗区内神经血管单元完整线粒体的平均体积变小且形态变圆;本研究还进一步提取并三维重建了内皮细胞中的线粒体并发现其形态和数量的改变。结论OFA流程可以快速获取神经血管单元或其中某个特定细胞类型,提供内皮细胞完整线粒体数量和体积的准确信息,为功能研究提供更准确全面的结构学证据。
基金supported by the National Natural Science Foundation of China(51401174)the National Social Science Foundation of China(16AYY015)the Scientific Research Funds of Southwest University of Political Science and Law(2015XZQN-32)
文摘A method for in situ preparation of fluorescent gold nanoclusters(Au NCs) with bovine serum albumin/montmorillonite composite powder(Au NC-BSA/MMT) was developed, and the products were used to detect latent fingermarks. In this work, Au NCs were "grown" both inside and on the surface of BSA/MMT clay using one-step reduction of HAu Cl4 by BSA. The as-prepared Au NC-BSA/MMT nanocomposites emit intensive red fluorescence under the excitation of UV-visible light and show stable chemical features and low toxicity. The obtained fluorescent powders were characterized by UV-visible absorption spectroscopy,fluorescence spectroscopy, infrared spectroscopy, transmission electron microscopy/high-resolution transmission electron microscopy, scanning electron microscopy, X-ray photoelectron spectroscopy and X-ray diffraction to depict their sizes, structural information and optical features. Given their environmentally friendly preparation, simple operation, low cost, efficient UVvisible radiation-dependent photoluminescence and good affinity with finger residues, the in situ synthesized Au NC-BSA/MMT nanocomposite powders were used as an alternative fluorescent developing reagent for developing latent fingermarks deposited on various object surfaces(such as glass, aluminum foil, painted metal, plastic products and weighing papers) for individual identification. As results, the developed fingermarks with clear patterns and satisfactory level-2(minutiae points) and level-3(sweat pores) ridge details were obtained. Notably, treated prints could be excited by red light and emitted near infrared fluorescence, which was beneficial to avoid background interference and reduce the damage caused by UV light. With the advantages of the simple preparation process and good enhancement performance for latent fingermarks, the proposed method might be used in the preparation of various fluorescent probes for detecting trace evidence in forensic sciences.
