Nonlinear focal modulation microscopy

TL;DR

Nonlinear focal modulation microscopy (NFOMM) is a new super-resolution imaging technique that extends spatial frequency bandwidth using phase modulation and high-intensity illumination, achieving 60 nm resolution.

Contribution

NFOMM introduces a nonlinear focal modulation approach with a simple, flexible setup that enhances resolution without complex modifications to existing microscopes.

Findings

Achieves 60 nm transverse resolution in fluorescent nanoparticle imaging.

Comparable performance to STED in imaging nuclear pore complexes.

Easy to implement as an add-on module for existing microscopes.

Abstract

Here we report nonlinear focal modulation microscopy (NFOMM) to achieve super-resolution imaging. Abandoning the previous persistence on minimizing the size of Gaussian emission pattern by directly narrowing (e.g. Minimizing the detection pinhole in Airyscan, Zeiss) or by indirectly peeling its outer profiles (e.g., Depleting the outer emission region in STED, stimulated emission microscopy) in pointwise scanning scenarios, we stick to a more general basis------ maximizing the system frequency shifting ability. In NFOMM, we implement a nonlinear focal modulation by applying phase modulations with high-intensity illumination, thereby extending the effective spatial-frequency bandwidth of the imaging system for reconstructing super-resolved images. NFOMM employs a spatial light modulator (SLM) for assisting pattern-modulated pointwise scanning, making the system single beam path while achieving a transverse resolution of 60 nm on imaging fluorescent nanoparticles. While exploring a relatively simple and flexible system, the imagingperformance of NFOMM is comparable with STED as evidenced in imaging nuclear pore complexes, demonstrating NFOMM is a suitable observation tool for fundamental studies in biology. Since NFOMM is implemented as an add-on module to an existing laser scanning microscope and easy to be aligned, we anticipate it will be adopted rapidly by the biological community.