阴极发光和特征性X射线发射稀土合核心/蛋白标签用于细胞表面受体的光谱显微镜分析
Sebastian Habermann1,2, Lukas R H Gerken1,2, Mathieu Kociak3
1Nanoparticle Systems Engineering Laboratory, Institute of Energy and Process Engineering, Department of Mechanical and Process Engineering, ETH Zurich, Sonneggstrasse 3, 8092, Zurich, Switzerland.
Small (Weinheim an der Bergstrasse, Germany)
|September 9, 2024
概括
研究人员开发了用于电子显微镜的新型纳米粒子标签,使得细胞内纳米级蛋白质的精确定位成为可能. 这些胺化纳米粒子提供多色成像能力,用于增强细胞超结构分析.
科学领域:
- 纳米技术纳米技术
- 生物物理学的生物物理.
- 显微镜的使用方法
背景情况:
- 细胞超结构和生物分子相互作用很难在纳米尺度上可视化.
- 电子显微镜 (EM) 提供高分辨率,但由于灰度图像,难以明确地定位生物分子.
- 现有的方法缺乏精确的,多颜色的标签,用于相关的纳米尺度成像.
研究的目的:
- 开发和演示基于纳米粒子的多式蛋白质标签,用于相关性阴极光发电电子显微镜 (CCLEM) 和能量分散式X射线光谱显微镜 (EDX-SM).
- 为了在细胞环境中实现纳米尺度生物分子的精确定位和识别.
- 建立电子光谱显微镜的多色标签策略.
主要方法:
- 合成了20nm以下的兰化物合的核心/外纳米粒子,其亮度得到了优化.
- 功能化纳米颗粒与叶酸 (terbium-doped) 和咖啡酸 (europium-doped) 用于特定的蛋白质向.
- 利用单粒子阴极发光 (CL) 和能量散射X射线 (EDX) 信号用于标签定位和HeLa细胞的识别.
主要成果:
- 从功能化纳米粒子中证明了精确的单粒子CL发射和独特的EDX信号.
- 通过使用叶酸和咖啡酸结合,在HeLa细胞中成功定位了表面受体.
- 使用EDX实现基于颜色的受体定位,实现了高分辨率 (2.78nm像素大小) 的快速成像 (<2分钟/μm2).
结论:
- 开发了一种用于先进电子显微镜技术的新型纳米粒子标签系统.
- 启用了多色可视化和细胞环境中的生物分子的精确定位.
- 开辟了纳米尺度成像和了解细胞内的分子相互作用的新途径.
更多相关视频
08:13Visualizing Subcellular Localization of a Protein in the Heart Using Quantum Dots-Mediated Immuno-Labeling Followed by Transmission Electron Microscopy
Published on: September 16, 2022
2.7K
16:16Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
Published on: September 13, 2013
15.2K
相关概念视频
03:31Labeling DNA Probes
8.1K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
8.1K
01:19Protein Dynamics in Living Cells
2.1K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.1K
01:14Photoluminescence: Applications
383
Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
383