DEVELOPMENT OF PROBES FOR MULTIPHOTON & EFTEM MICROSCOPIES

我正在开发一种既可以用于多光子的探测器 激发荧光显微镜与能量过滤透射法 电子显微镜。将这两个显微镜集成在一起可以提供 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大的 减少离焦对活体标本研究的优势 激发，使生活的三维组织 由于减少了光毒性，组织将得到更好的研究。在……里面 能量过滤透射式电子显微镜小得多 免疫金标记物可以与标准的、透过性的 电子显微镜。我最近证明了140万 纳霍金可以清楚地显示在标记的细胞核内 样品，而5MN通常是最小尺寸 常规的透射电子显微镜。使用较小的 探针减少了空间位阻的问题，使标签能够 更好地渗透到细胞骨架的基质中。这个 这两种显微镜的特点相结合可能会提供很多东西 细胞生物学家，允许高分辨率，相关 体内分析后细胞学结构的可视化 活体标本的动力学。这种集成的方法是 特别强大，当与冷冻技术相结合时。的目标是 这些项目将探索各种组合的标记将被 用于集成显微镜。

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.