DEVELOPMENT OF PROBES FOR MULTIPHOTON & EFTEM MICROSCOPIES

多光子探针的开发

• 批准号: 6117269
• 负责人: MAREK MALECKI
• 金额: $ 1.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6117269
• 关键词: bioimaging /biomedical imaging; biomedical resource; biotechnology; human tissue; lasers; microscopy

I am developing probes that may be used with both multiphoton excitation fluorescence microscopy and energy-filtered transmission electron microscopy. Integrating these two microscopies can provide us with a powerful tool for studying intracellular processes. Multiphoton excitation fluorescence microscopy provides considerable advantages for studying live specimens because of reduced out-of-focus excitation, enabling the thi-ee-dimensional organization of live tissue to be better studied because of reduced phototoxicity. In energy filtering transmission electron microscopy much smaller immuno-gold labels can be used compared with standard, transmission electron microscopy. I have recently demonstrated that 1.4 mn nahogold can be clearly visualized within the nucleus of a labeled specimen, compared with 5mn which is generally the minimum size for conventional transmission electron microscopy. The use of smaller probes reduces problems of steric hindrance, allowing the label to infiltrate better into the matrix of the cytoskeleton. The combination of featurts of both microscopies may have much to offer the cell biologist, allowing high-resolution, correlative visualization of cytological structure following analysis of in vivo dynamics of the living specimen. This integrated approach is particularly powerful, when combined with cryotechniques. The goal of these project will be to explore various combinations of markers to be used for integrated microscopy.

我正在开发的探测器，可用于多光子 激发荧光显微术和能量过滤透射 电镜 整合这两种显微镜可以提供 为我们提供了一个研究细胞内过程的强大工具。 多光子激发荧光显微镜提供了相当多的 由于减少了散焦，因此有利于研究活体标本 激发，实现现场的三维组织 因为减少了光毒性，组织将被更好地研究。 在 能量过滤透射电子显微镜 免疫金标记可用于与标准、透射 电镜 我最近证明了140万 Nahogold可以在标记的细胞核内清楚地可视化。 试样，相比之下，5mn通常是最小的尺寸， 常规透射电子显微镜。 使用较小的 探针减少了空间位阻的问题，允许标记物 更好地渗透到细胞骨架的基质中。 的 结合两种显微镜的特点， 细胞生物学家，允许高分辨率，相关 体内分析后的细胞学结构可视化 活标本的动力学。 这种综合方法是 当与冷冻技术相结合时，特别强大。 的目标 这些项目将探索标记的各种组合， 用于集成显微镜。