Optimization and characterization of toroidal foci for super-resolution fluorescence microscopy: tutorial

Single-molecule localization microscopy (SMLM) has become an essential tool to investigate phenomena at the nanoscale. Among the different SMLM approaches, methods that interrogate the molecular position with an intensity minimum, such as minimal emission fluxes (MINFLUX) or the more recent raster s...

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Detalles Bibliográficos
Autores: Lopez, Lucía Fernanda, Masullo, Luciano Andrés, Szalai, Alan Marcelo, Edorna, Florencia Belen, Choque, Florencia Daniela, Caprile, Fernando, Stefani, Fernando Daniel
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:Argentina
Institución:Consejo Nacional de Investigaciones Científicas y Técnicas
Repositorio:CONICET Digital (CONICET)
Idioma:inglés
OAI Identifier:oai:ri.conicet.gov.ar:11336/228958
Acceso en línea:http://hdl.handle.net/11336/228958
Access Level:acceso abierto
Palabra clave:Fluorescence
Single Molecule Localization Microscopy
MINFLUX
https://purl.org/becyt/ford/1.3
https://purl.org/becyt/ford/1
Descripción
Sumario:Single-molecule localization microscopy (SMLM) has become an essential tool to investigate phenomena at the nanoscale. Among the different SMLM approaches, methods that interrogate the molecular position with an intensity minimum, such as minimal emission fluxes (MINFLUX) or the more recent raster scanning a minimum of light (RASTMIN), stand out for reaching true molecular resolution. To implement these methods, the phase of the excitation beam needs to be modulated to obtain a focus with a central minimum, i.e., a so-called toroidal or doughnut-shaped focus. In this tutorial,we explain the basis and experimental tricks to generate and optimize such beams, particularly in raster-scanning microscopes.