组织固定对结构部件的光学特性的影响,通过非线性显微镜成像评估
M Andrea Markus1, Daniele P Ferrari1, Frauke Alves1,2,3
1Translational Molecular Imaging Group, Max-Planck Institute for Multidisciplinary Sciences, Hermann-Rein-Str. 3, 37075 Göttingen, Germany.
Biomedical optics express
|October 6, 2023
概括
诸如甲素这样的组织固定方法会改变蛋白质结构,影响非线性显微镜信号. 在研究中,选择正确的保存技术对于准确的原和肌素可视化至关重要.
科学领域:
- 生物医学成像技术 生物医学成像技术
- 历史学 历史学 历史学
- 分子生物学分子生物学
背景情况:
- 组织固定对于保存生物样本至关重要.
- 化学固定剂,如甲素,可以化学修改组织成分,如原蛋白和肌素.
- 非线性光学显微镜技术,包括第二和第三和生成 (SHG和THG),对分子结构敏感.
研究的目的:
- 评估不同组织保护方法对关键结构蛋白的非线性光学特性的影响.
- 确定固定技术如何影响来自原蛋白,脂质滴和肌肉素的SHG和THG信号.
- 为了指导选择最佳的固定方法,用于先进的成像和组织蛋白质的量化.
主要方法:
- 使用非线性双光子显微镜 (NL2PM) 来评估来自小鼠组织的SHG和THG排放.
- 对比了各种固定方法的影响,包括甲素和冷.
- 分析了原纤维参数 (密度,分支点) 和髓微结构的变化.
主要成果:
- 甲林固定显著改变了原纤维参数,特别是在原稀疏区域,如脏.
- 冷导致肌肉素SHG信号的显著减少.
- 原蛋白,脂质滴和肌肉蛋白的非线性光学能力被保存技术明显影响.
结论:
- 组织固定方法对结构蛋白的非线性光学信号有着深刻的影响.
- 甲素固定可以扭曲原结构,而冷会减少与肌素相关的信号.
- 选择适当的固定方法对于使用NL2PM准确可视化和量化蛋白质至关重要,有助于疾病研究.
更多相关视频
相关概念视频
01:03Fixation and Sectioning
4.4K
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...
4.4K
01:18Imaging Biological Samples with Optical Microscopy
4.8K
Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
4.8K
01:20Preparation of Samples for Electron Microscopy
5.5K
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...
5.5K
01:17Studying the Cytoskeleton
6.3K
The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
6.3K