Fernando Stefani

Luciano A. Masullo 1,  Alan M. Szalai 1,  Lucía F. López 1, Mauricio Pilo-Pais 2, Guillermo P. Acuna 3, Fernando D. Stefani 1

1- Center for Bionanoscience Research (CIBION), Buenos Aires, Argentina, 2- University of Fribourg, Switzerland, 3- University of Fribourg, Switzerland

Here, we present RASTMIN (RASTer scanning MINima), a single-molecule localization method that delivers a localization precision equivalent to MINFLUX but is easily implementable on any laser-scanning (confocal) microscope with few modifications [1]. We present the working principle, simulations, experimental set-up, and performance demonstrations through single-molecule localization measurements and super-resolution imaging of DNA-origami structures. RASTMIN is fully compatible with lifetime measurements and with any other laser-scanning fluorescence microscopy techniques such as two-photon microscopy. In addition, we present a simpler conceptual framework to understand this method as well as other reported methods of the same kind, such as MINFLUX, orbital tracking or MINSTED [2].

[1] Masullo, L. A.; Szalai, A. M.; Lopez, L. F.; Pilo-Pais, M.; Acuna, G. P.; Stefani, F. D. An Alternative to MINFLUX That Enables Nanometer Resolution in a Confocal Microscope. Light Sci. Appl. 2022, 11 (1), 199. https://doi.org/10.1038/s41377-022-00896-4.[2] Masullo, L. A.; Lopez, L. F.; Stefani, F. D. A Common Framework for Single-Molecule Localization Using Sequential Structured Illumination. Biophys. Reports 2022, 2 (1), 100036. https://doi.org/10.1016/j.bpr.2021.100036.