Compared with other imaging techniques,fluorescence microscopy has become an essential tool to study cell biology due to its high compatibility with living cells.Owing to the resolution limit set by the diffraction of light,fluorescence microscopy could not resolve the nanostructures in the range of<200 nm.Recently,many techniques have been emerged to overcome the diffraction barrier,providing nanometer spatial resolution.In the course of development,the progress in fluorescent probes has helped to promote the development of the high-resolution fluorescence nanoscopy.Here,we describe the contributions of the fluorescent probes to far-field super resolution imaging,focusing on concepts of the existing super-resolution nanoscopy based on the photophysics of fluorescent nanoprobes,like photoswitching,bleaching and blinking.Fluorescent probe technology is crucial in the design and implementation of super-resolution imaging methods.
Recently,we theoretically demonstrate that utilization of silica nanobeads co-doped with Cy3 and Cy5 molecules instead of single dye molecules asfluorescent labels can enable optical resolutions far beyond the diffraction-limit.Here,we show that by combining the 4Pi microscopy and the novelfluorescent label,it is possible to completely suppress the sidelobes in 4Pi focal spot and significantly enhance the optical resolution in the axial direction.
