Abstract
Keywords
1 Introduction
The resolution of common fluorescence microscopes (wide-field or confocal microscopes) is limited by the diffraction of light, known as the Abbe limit. The attainable resolution is given by the full-width at half-maximum (FWHM) of the point spread function (PSF) of the beam at the focus of the objective. A high numerical aperture () objective with visible light () can theoretically reach a resolution of and for the confocal and wide field, respectively, whereas the experimental resolution is generally in the range of to 250 nm due to the sample optical properties and beam imperfections. Super-resolution fluorescence microscopy (SRM) permits us to beat the diffraction limit, and it obtains images with a higher resolution, from 100 nm to as low as 20 nm or, in some cases, even lower, with few nanometer localization in some cases. This is a resolution/localization possible only by electron scanning probe microscopes. SRM’s impact in life science, chemical, and physical sciences has been recognized by the Nobel Prize for Chemistry in 2014,1 and it has revolutionized many areas of cellular microscopy2 and even virology.3,4
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