Fundamental precision limits of fluorescence microscopy: a perspective on MINFLUX
Year: 2024
Authors: Rosati M., Parisi M., Gianani I., Barbieri M., Cincotti G.
Autors Affiliation: Univ Roma Tre, Dipartimento Ingn Civile Informat & Tecnol Aeronau, Via Vito Volterra 62, I-00146 Rome, Italy; Univ Roma Tre, Dipartimento Sci, Via Vasca Navale 84, I-00146 Rome, Italy; Ist Nazl Ottica CNR, Largo E Fermi 6, I-50125 Florence, Italy.
Abstract: Over the past years, fluorescence microscopy (FM) has steadily progressed in increasing the localization precision of fluorescent emitters in biological samples and led to new claims, whose rigorous validation remains an outstanding problem. We present a novel, to the best of our knowledge, multi-parameter estimation framework that captures the full complexity of a single-emitter FM localization experiment. We showcase our method with Minimum Flux (MINFLUX) microscopy, among the highest-resolution approaches, demonstrating that (i) the localization precision can be increased only by turning the illumination intensity up, thus increasing the risk of photo-bleaching, and it is independent from the beams’ separation, and (ii) in presence of background noise, the localization precision decreases with the beams’ separation. Finally, we apply our method to Minimum Flux Stimulated Emission Depletion (MINSTED) microscopy, showing that a reduction of the beam width can provide similar performance to MINFLUX. (c) 2024 Optica Publishing Group. All rights, including for text and data mining (TDM), Artificial Intelligence (AI) training, and similar technologies, are reserved.
Journal/Review: OPTICS LETTERS
Volume: 49 (17) Pages from: to:
More Information: Ministero dell’Universita e della Ricerca 816000-2022-SQUID – CUP F83C22002390007 (Young Researchers); H2020 Future and Emerging Technologies (STORMYTUNE 899587).KeyWords: Fisher Information; LocalizationDOI: 10.1364/OL.530358