Discriminating single-molecule binding events from diffraction-limited fluorescence.

ABSTRACT: Single-molecule localization microscopy enables high-resolution imaging of molecular interactions, but discriminating molecular binding types has traditionally relied on complex strategies, such as multiple dyes, time-division techniques, or kinetic analysis, that are asynchronous, invasive, or time-consuming. Here, we uncover previously overlooked spatiotemporal information embedded within diffraction-limited fluorescence, enabling synchronous classification of individual binding event videos using only a single fluorescent dye. Building on this insight, we propose a Temporal-to-Context Convolutional Neural Network (T2C CNN), which integrates long-term spatial convolutions, shallow cross-connected blocks, and a pooling-free structure to enhance contextual representation while preserving f

SUBMITTER: Yin Y 

PROVIDER: S-EPMC12595110 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature