Zhi Yanan, Wang Benquan, Yao Xincheng
Department of Bioengineering, University of Illinois at Chicago, Chicago, IL.
Department of Bioengineering, University of Illinois at Chicago, Chicago, IL; Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, IL.
Crit Rev Biomed Eng. 2015;43(4):297-322. doi: 10.1615/CritRevBiomedEng.2016016445.
Light microscopy plays a key role in biological studies and medical diagnosis. The spatial resolution of conventional optical microscopes is limited to approximately half the wavelength of the illumination light as a result of the diffraction limit. Several approaches-including confocal microscopy, stimulated emission depletion microscopy, stochastic optical reconstruction microscopy, photoactivated localization microscopy, and structured illumination microscopy-have been established to achieve super-resolution imaging. However, none of these methods is suitable for the super-resolution ophthalmoscopy of retinal structures because of laser safety issues and inevitable eye movements. We recently experimentally validated virtually structured detection (VSD) as an alternative strategy to extend the diffraction limit. Without the complexity of structured illumination, VSD provides an easy, low-cost, and phase artifact-free strategy to achieve super-resolution in scanning laser microscopy. In this article we summarize the basic principles of the VSD method, review our demonstrated single-point and line-scan super-resolution systems, and discuss both technical challenges and the potential of VSD-based instrumentation for super-resolution ophthalmoscopy of the retina.
光学显微镜在生物学研究和医学诊断中起着关键作用。由于衍射极限，传统光学显微镜的空间分辨率被限制在照明光波长的大约一半左右。为了实现超分辨率成像，已经建立了几种方法，包括共聚焦显微镜、受激发射损耗显微镜、随机光学重建显微镜、光激活定位显微镜和结构光照明显微镜。然而，由于激光安全问题和不可避免的眼球运动，这些方法都不适用于视网膜结构的超分辨率检眼镜检查。我们最近通过实验验证了虚拟结构检测（VSD）作为扩展衍射极限的替代策略。VSD无需结构照明的复杂性，提供了一种简单、低成本且无相位伪影的策略，以在扫描激光显微镜中实现超分辨率。在本文中，我们总结了VSD方法的基本原理，回顾了我们展示的单点和线扫描超分辨率系统，并讨论了技术挑战以及基于VSD的仪器用于视网膜超分辨率检眼镜检查的潜力。
