Super-Resolution Microscopy Based on the Inherent Fluctuations of Dye Molecules

TL;DR

This paper introduces a new super-resolution microscopy method that leverages the natural fluctuations of dye molecules, enabling 3D and multicolor imaging without complex sample preparation or specialized optical systems.

Contribution

It presents a simple, robust super-resolution technique using dye fluctuations and an array of single-photon detectors, eliminating the need for engineered blinking or complex setups.

Findings

Achieved 3D and multicolor super-resolution imaging.

Utilized natural dye fluctuations as a contrast mechanism.

Demonstrated compatibility with standard confocal architecture.

Abstract

Fluorescence microscopy is a critical tool across various disciplines, from materials science to biomedical research, yet it is limited by the diffraction limit of resolution. Advanced super-resolution techniques such as localization microscopy and stimulated-emission-depletion microscopy often demand considerable resources. These methods depend heavily on elaborate sample-staining, complex optical systems, or prolonged acquisition periods, and their application in 3D and multicolor imaging presents significant experimental challenges. In the current work, we provide a complete demonstration of a widely accessible super-resolution imaging approach capable of 3D and multicolor imaging. We replace the confocal pinhole with an array of single-photon avalanche diodes and use the microsecond-scale fluctuations of dye molecules as a contrast mechanism. This contrast is transformed into a super-resolved image using a robust and deterministic algorithm. Our technique utilizes natural fluctuations inherent to organic dyes, thereby it does not require engineering of the blinking statistics. Our robust, versatile super-resolution method opens the way to next-generation multimodal imaging and facilitates on-demand super-resolution within a confocal architecture.