Principles of microscopy for ophthalmologists

Abstract

This short review begins with the theories of Airy, Rayleigh and Abbe on microscope resolution. Next, the principal developments in microscopy in the last half‐century are examined for relevance to ophthalmology: confocal microscopy, photoactivation light microscopy (PALM), stochastic optical reconstruction microscopy (STORM), stimulated emission depletion (STED), structured illumination (SI), 2‐photon and multiphoton excitation microscopy with a focused beam. Except for confocal, these are difficult to apply to the eye in vivo, as are the interference methods available in microscopes. However, interferometry in the form of coherence tomography is now a major ophthalmic method which has diverged from microscopy. Multiphoton excitation microscopy with an unfocussed beam is a new, low‐damage microscope method so‐far not exploited in ophthalmoscopy. The Mesolens, which throws off the historic limitations in microscopy set by the human eye, is described as a possible future aid to ophthalmology of the anterior eye.

摘要

此短篇综述从Airy、Rayleigh和Abbe关于显微镜分辨率的理论开始。随后, 我们阐述过去半个世纪显微镜的发展主线与眼科的相关性: 共聚焦显微镜、光激活显微镜 (PALM) 、随机光学重建显微镜 (STORM) 、受激发射耗尽显微镜 (STED) 、结构照明显微镜 (SI) 、聚焦光束的双光子和多光子激发显微镜。除了共聚焦显微镜外, 这些显微镜都很难在体内应用, 显微镜中可用的干涉方法也是如此。不过, 相干断层扫描形式的干涉测量法目前已成为一种主要的眼科方法, 并已从显微镜中分离出来。使用非聚焦光束的多光子激发显微镜是一种新型、低损伤的显微镜方法, 迄今为止尚未在眼科显微镜中得到应用。Mesolens 打破了人眼显微镜的历史局限性, 未来可能成为眼科前节的辅助性手段。