Far‐Field Nanoscopy with Reversible Chemical Reactions

Abstract

Revolution in resolution: Abbe's resolution limit has been overcome in fluorescence microscopy by using light‐driven processes to switch the emission of fluorophores on and off. Alternatively, chemical reactions can be used, for example the coordination of Cu2+ ions to a fluorescent probe for the stochastic switching between spectroscopic states.

Single-molecule chemical reaction reveals molecular reaction kinetics and dynamics

“…1a) 20 . This process can be reproduced again and again for many times due to the reversibility of this redox process ( Supplementary Figs 1 and 2) [21][22][23] .…”

“…To get more clues about reactivity dynamic fluctuation of single dye molecules, we used the fitting of the emission point spread function 40,41 , the basis for optical super-resolution imaging methods 23,42,43 , to 'track' the movement of single dye molecules in SiO 2 nanospheres. In this way, we can map the location variation of a free single dye molecule in situ inside the cavity of a hollow SiO 2 nanosphere during the repeated redox process (Supplementary Methods and Discussion; Supplementary Figs 11 and 12).…”

Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging

“…Examples in the second category include dark-to-bright fluorescent proteins, such as PA-GFP 28 and PA-mCherry 29 , color-converting fluorescent proteins, such as PS-CFP 30 , EosFP 31,32 , and Dendra2 33 , and push-pull fluorogen dyes 18 , although in some cases photoactivatable fluorescent proteins may also undergo reversible photoswitching once photoactivated. In addition to photoswitchable fluorophores, probes that can be turned on and off by non-optical means, such as reversible ligand binding 34 or by fluorescent quenchers 19 , have also been used for localization-based super-resolution fluorescence microscopy (Fig. 1b).…”

Super-Resolution Molecular and Functional Imaging of Nanoscale Architectures in Life and Materials Science

“…Recently, the switching of fluorescent states without the requirement for light illumination has been achieved by chemical (Schwering et al, 2011; Vaughan et al, 2012) and enzymatic (Lee et al, 2013) reactions (Figure 2E).…”