Jester James V, Ward Brant R, Takashima Akira, Gatlin Joel, Garcia J Victor, Cavanagh H Dwight, Petroll W Matthew
Department of Ophthalmology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390-9057, USA.
Ocul Surf. 2004 Jan;2(1):10-20. doi: 10.1016/s1542-0124(12)70020-4.
This paper reviews new developments in microscopy that combine gene transfer technology, multiphoton confocal fluorescence microscopy, live cell imaging and digital imaging techniques that provide unique insights into the complex physiological processes involved in tissue function at the cellular and subcellular level. The evolution of this novel, new technology is discussed with particular attention to earlier achievements in noninvasive ocular surface imaging. The practical basis of confocal microscopy, multiphoton confocal fluorescence microscopy, and the vital fluorescent labeling of cells in living tissues are also discussed. Additionally, one application using retroviral gene transfer to express enhanced green fluorescent protein in living wound healing fibroblasts is presented as an example of how living biology can be studied in situ in four dimensions (x, y, z, time).
本文综述了显微镜技术的新进展，这些进展结合了基因转移技术、多光子共聚焦荧光显微镜、活细胞成像和数字成像技术，能够在细胞和亚细胞水平上为组织功能所涉及的复杂生理过程提供独特的见解。本文讨论了这项新颖的新技术的发展，特别关注非侵入性眼表成像的早期成果。还讨论了共聚焦显微镜、多光子共聚焦荧光显微镜的实际基础以及活组织中细胞的活体荧光标记。此外，还介绍了一个利用逆转录病毒基因转移在活的伤口愈合成纤维细胞中表达增强型绿色荧光蛋白的应用实例，以此说明如何在四维（x、y、z、时间）原位研究活体生物学。
