Hacking the optical diffraction limit: Review on recent developments of fluorescence nanoscopy

Abstract

Subject to the diffraction limit, the resolution of conventional optical microscopy is constrained to about 200 and 500 nm in the lateral and axial planes, respectively. The advantage of optical microscopy in the life sciences over electronic microscopy, especially fluorescence microscopy, drives scientists to develop novel "hacks" to reach nanoscale resolutions by optical means. In this review, three aspects of the techniques are discussed: (1) lateral super-resolution; (2) axial super-resolution; (3) super-resolution in three dimensions. The principles of how the methods achieve the cross-barrier resolution are discussed, and recent advances in current techniques are described. With these methods, the use of fluorescence microscopy is growing quickly toward a new era: fluorescence nanoscopy that will reveal 2 orders of magnitude more information on cellular structure and dynamics. optical nanoscopy, fluorescence microscopy, diffraction limit Citation:Ding Y C, Xi P, Ren Q S. Hacking the optical diffraction limit: Review on recent developments of fluorescence nanoscopy.

Three-dimensional multimodal sub-diffraction imaging with spinning-disk confocal microscopy using blinking/fluctuating probes

“…In SD-BaLM processing, the localization precision could be decreased to the nanometer scale, achieving a resolution of less than 30 nm. The axial resolution, however, is also constrained by the PSF of the SD confocal system [15,48].…”

Advanced Biological Imaging for Intracellular Micromanipulation: Methods and Applications

“…Similar to CFM, WFFM suffer from a diffraction barrier. The resolution of WFFM can be expressed as λ 2NA (lateral) and 2λ NA 2 (axial) [98], where λ is the wavelength of the emission light, and NA is the numerical aperture of the objective lens. Even for a WFFM with a high-NA objective lens, its resolution hardly reaches 230 nm laterally and 800 nm axially because the visible light wavelength varies from 390 to 780 nm [99].…”

Computational methods in super-resolution microscopy

“…The past few decades have testified to the rapid development of various super-resolution microscopy techniques (Ding et al, 2011;Gu et al, 2014;Gao et al, 2016;Yang et al, 2016a;Yu et al, 2016). Because the process of optical imaging is the convolution of the original object and the point spread function (PSF) of the system, the wave nature of light limits the resolution of conventional optical microscopy.…”