对死后人类大脑组织的固定和染色的协议优化为聚焦离子束扫描电子显微镜
Matthew J Maier1, David A Lewis1, Zachary Freyberg2
1University of Pittsburgh, School of Medicine, Department of Psychiatry, Pittsburgh, PA 15213, USA.
STAR protocols
|January 16, 2026
概括
这项研究详细介绍了一种准备死后人类大脑组织的协议,用于使用聚焦离子束扫描电子显微镜 (FIB-SEM) 的高分辨率体积电子显微镜 (vEM). 该方法确保了对详细的3D重建的优异超结构性保存.
科学领域:
- 神经科学是一个神经科学.
- 显微镜的使用方法
- 生物技术是生物技术.
背景情况:
- 死亡后人脑组织的高分辨率成像对于理解神经系统疾病至关重要.
- 现有的协议往往难以为先进的显微镜技术保留微妙的超结构.
研究的目的:
- 提出一个强大的协议,用于生成经验丰富的体积电子显微镜 (vEM) 优化的人类脑组织样本.
- 为了使人类大脑组织的详细的三维 (3D) 超结构重建.
主要方法:
- 大脑组织的沉浸固定.
- 切割和长期冷保存技术.
- 增强对比染色用于改进的VEM成像.
主要成果:
- 该协议产生了死后人类大脑样本,具有高质量的超结构性保存.
- 样品适用于高分辨率聚焦离子束扫描电子显微镜 (FIB-SEM).
- 能够对细胞和亚细胞特征进行密集和有针对性的3D重建.
结论:
- 该协议提供了一种可靠的方法,用于为vEM准备人体死后脑组织.
- 简化了详细的超结构分析和人类大脑的3D重建.
- 通过改进成像能力,推进神经生物学和神经病理学的研究.
相关概念视频
Preparation of Samples for Electron Microscopy
6.8K
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
6.8K
Fixation and Sectioning
7.4K
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
7.4K


