Extending resolution within a single imaging frame
Esley Torres, Raúl Pinto‐Cámara, Alejandro Linares, Damián Martínez, Víctor Abonza, Eduardo Brito‐Alarcón, Carlos Calcines-Cruz, Erick G. Valdés-Galindo, David Torres, Martina Jabloñski, Héctor H. Torres‐Martínez, José Luis Martínez, Haydeé O. Hernández, José P. Ocelotl-Oviedo, Yasel Garcés Suárez, Marco Barchi, Rocco D’Antuono, Ana Bošković, Joseph Dubrovsky, Alberto Darszon, Mariano G. Buffone, Roberto Morales, Juan Manuel Rendón-Mancha, Christopher D. Wood, Armando Hernández-García, Diego Krapf, Álvaro H. Crevenna, Adán Guerrero
Abstract
The resolution of fluorescence microscopy images is limited by the physical properties of light. In the last decade, numerous super-resolution microscopy (SRM) approaches have been proposed to deal with such hindrance. Here we present Mean-Shift Super Resolution (MSSR), a new SRM algorithm based on the Mean Shift theory, which extends spatial resolution of single fluorescence images beyond the diffraction limit of light. MSSR works on low and high fluorophore densities, is not limited by the architecture of the optical setup and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image stacks, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other SRM approaches. Along with its wide accessibility, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.