Perspective: An outlook on fluorescence tracking

TL;DR

This perspective reviews the evolution of fluorescence tracking techniques for single molecules, comparing methods, discussing emerging physics-inspired and AI-based approaches, and highlighting challenges and opportunities for improving resolution and efficiency.

Contribution

It provides a comprehensive overview of the development, current state, and future directions of fluorescence tracking methods in single-molecule studies.

Findings

Comparison of various fluorescence tracking methods

Discussion of emerging physics-inspired and AI techniques

Identification of challenges and opportunities for future improvements

Abstract

Tracking single fluorescent molecules has offered resolution into dynamic molecular processes at the single-molecule level. This perspective traces the evolution of single-molecule tracking, highlighting key developments across various methodological branches within fluorescence microscopy. We compare the strengths and limitations of each approach, ranging from conventional widefield offline tracking to real-time confocal tracking. In the final section, we explore emerging efforts to advance physics-inspired tracking techniques, a possibility for parallelization and artificial intelligence, and discuss challenges and opportunities they present toward achieving higher spatiotemporal resolution and greater computational and data efficiency in next-generation single-molecule studies.