Real-time super-resolution imaging by high-speed fluorescence emission difference microscopy

Abstract Overcoming the resolution limit of conventional optics is regarded as the most important issue in optical imaging science and technology. Although hyperlenses, super-resolution imaging devices based on highly anisotropic dispersion relations that allow the access of high-wavevector components, have recently achieved far-field sub-diffraction imaging in real-time, the previously demonstrated devices have suffered from the extreme difficulties of both the fabrication process and the non-artificial objects placement. This results in restrictions on the practical applications of the hyperlens devices. While implementing large-scale hyperlens arrays in conventional microscopy is desirable to solve such issues, it has not been feasible to fabricate such large-scale hyperlens array with the previously used nanofabrication methods. Here, we suggest a scalable and reliable fabrication process of a large-scale hyperlens device based on direct pattern transfer techniques. We fabricate a 5 cm × 5 cm size hyperlenses array and experimentally demonstrate that it can resolve sub-diffraction features down to 160 nm under 410 nm wavelength visible light. The array-based hyperlens device will provide a simple solution for much more practical far-field and real-time super-resolution imaging which can be widely used in optics, biology, medical science, nanotechnology and other closely related interdisciplinary fields.

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Real-time super-resolution imaging using a single sensor

2015 1st URSI Atlantic Radio Science Conference (URSI AT-RASC) ◽

10.1109/ursi-at-rasc.2015.7302946 ◽

2015 ◽

Author(s):

Lianlin Li ◽

Henxin Ruan ◽

Fang Li ◽

Tiejun Cui

Keyword(s):

Real Time ◽

Super Resolution ◽

Single Sensor ◽

Resolution Imaging

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FluoSim: simulator of single molecule dynamics for fluorescence live-cell and super-resolution imaging of membrane proteins

10.1101/2020.02.06.937045 ◽

2020 ◽

Cited By ~ 3

Author(s):

Matthieu Lagardère ◽

Ingrid Chamma ◽

Emmanuel Bouilhol ◽

Macha Nikolski ◽

Olivier Thoumine

Keyword(s):

Real Time ◽

Protein Dynamics ◽

Single Molecule ◽

Imaging Techniques ◽

Simulated Data ◽

Super Resolution ◽

Molecular Complexes ◽

Live Cell ◽

Single Molecule Level ◽

Resolution Imaging

AbstractFluorescence live-cell and super-resolution microscopy methods have considerably advanced our understanding of the dynamics and mesoscale organization of macro-molecular complexes that drive cellular functions. However, different imaging techniques can provide quite disparate information about protein motion and organization, owing to their respective experimental ranges and limitations. To address these limitations, we present here a unified computer program that allows one to model and predict membrane protein dynamics at the ensemble and single molecule level, so as to reconcile imaging paradigms and quantitatively characterize protein behavior in complex cellular environments. FluoSim is an interactive real-time simulator of protein dynamics for live-cell imaging methods including SPT, FRAP, PAF, and FCS, and super-resolution imaging techniques such as PALM, dSTORM, and uPAINT. The software, thoroughly validated against experimental data on the canonical neurexin-neuroligin adhesion complex, integrates diffusion coefficients, binding rates, and fluorophore photo-physics to calculate in real time the distribution of thousands of independent molecules in 2D cellular geometries, providing simulated data of protein dynamics and localization directly comparable to actual experiments.

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