Trade-offs between structural integrity and acquisition time in stochastic super-resolution microscopy techniques.

The applicability of widefield stochastic microscopy, such as PALM or STORM, is limited by their long acquisition times. Images are produced from the accumulation of a large number of frames that each contain a scarce number of super-resolved localizations. We show that the random and uneven distribution of localizations leads to a specific type of trade-off between the spatial and temporal resolutions. We derive analytical predictions for the minimal time required to obtain a reliable image at a given spatial resolution. We find that the image completion time scales logarithmically with the ratio of the image size to the spatial resolution volume, with second order corrections due to spurious localization within the background noise. We validate our predictions against experimental localization sequences of labeled microtubule filaments obtained by STORM. Our theoretical framework makes it possible to compare the efficiency of emitters, define optimal labeling strategies, and allow implementation of a stopping criterion for data acquisitions that can be performed using real-time monitoring algorithms.

Correlated Electrochemical and Optical Detection Reveals the Chemical Reactivity of Individual Silver Nanoparticles.

Electrochemical (EC) impacts of single nanoparticles (NPs) on an ultramicroelectrode are coupled with optics to identify chemical processes at the level of individual NPs. While the EC signals characterize the charge transfer process, the optical monitoring gives a complementary picture of the transport and chemical transformation of the NPs. This is illustrated in the case of electrodissolution of Ag NPs. In the simplest case, the optically monitored dissolution of individual NPs is synchronized with individual EC spikes. Optics then validates in situ the concept of EC nanoimpacts for sizing and counting of NPs. Chemical complexity is introduced by using a precipitating agent, SCN(-), which tunes the overall electrodissolution kinetics. Particularly, the charge transfer and dissolution steps occur sequentially as the synchronicity between the EC and optical signals is lost. This demonstrates the level of complexity that can be revealed from such electrochemistry/optics coupling.

Deciphering the Elementary Steps of Transport-Reaction Processes at Individual Ag Nanoparticles by 3D Superlocalization Microscopy.

Transport-reaction processes at individual Ag nanoparticles (NPs) are studied using electrochemistry coupled with in situ 3D light scattering microscopy. Electrochemistry is used to trigger a (i) diffusiophoretic transport mode capable of accelerating and preconcentrating NPs toward an electrode and (ii) subsequent diffusion-controlled oxidation of NPs. Individual NP dissolution rate, analyzed using optical modeling, suggests the intervention of insoluble products. New insights into diverse NPs behaviors highlight the strength of coupled optical-electrochemical 3D microscopies for single-NP studies.

Stochastic 3D optical mapping by holographic localization of Brownian scatterers.

We present a wide-field microscopy technique for the 3D mapping of optical intensity distributions using Brownian gold nanopar-ticles as local probes, which are localized by off-axis holography. Fast computation methods allow us to localize hundreds of particles per minute with accuracies as good as 3 × 3 × 10nm³ for immobilized particles. Factors limiting this accuracy are discussed and the possibilities of the technique are illustrated through the 3D optical mapping of an evanescent and a propagative wave. Our results pave the way for a new stochastic imaging technique, well adapted to subwavelength optical characterization in water-based systems.