DEVELOPMENT OF PROBES FOR MULTIPHOTON & EFTEM MICROSCOPIES

多光子探针的开发

• 批准号: 6278464
• 负责人: MAREK MALECKI
• 金额: $ 0.71万
• 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: 1998-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6278464
• 关键词: 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.

我正在开发可与多光子一起使用的探针 激发荧光显微镜和能量过滤透射 电子显微镜。 整合这两种显微镜可以提供 为我们提供了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 由于失焦减少，有利于研究活体标本 激发，使现场的三维组织成为可能 由于光毒性降低，因此可以更好地研究组织。 在 能量过滤透射电子显微镜小得多 可以使用免疫金标记与标准、透射比 电子显微镜。 我最近证明了 140 万 nahogold 可以在标记的细胞核内清晰可见 样本，与 5mn 相比，通常是最小尺寸 传统的透射电子显微镜。 使用较小的 探针减少了空间位阻问题，使标记能够 更好地渗透到细胞骨架基质中。 这 两种显微镜功能的结合可能会带来很多好处 细胞生物学家，允许高分辨率、相关性 体内分析后细胞学结构的可视化 活体标本的动力学。 这种综合方法是 与冷冻技术结合时特别强大。 目标是 这些项目将探索标记的各种组合 用于集成显微镜。