Adán Guerrero

Esley Torres García1,2, Raúl Pinto Cámara1,2, Alejandro Linares2,3, Damián Martínez2, Víctor Abonza2, Eduardo Brito-Alarcón2, Carlos Calcines-Cruz4, Gustavo Valdés Galindo5, David Torres2, Martina Jabloñski6, Héctor H. Torres-Martínez7, José L. Martínez8, Haydee O. Hernández9, José P. Ocelotl-Oviedo2, Yasel Garcés2,8, Marco Barchi10, Rocco D'Antuono11, Ana Bošković12, Joseph G. Dubrovsky7, Alberto Darszon8, Mariano G. Buffone6, Roberto Rodríguez Morales13, Juan Manuel Rendon-Mancha1 Christopher D. Wood2, Armando Hernández-García5, Diego Krapf14,  Álvaro H. Crevenna12 and Adán Guerrero2

1-      Centro de Investigación en Ciencias, Instituto de Investigación en Ciencias Básicas y Aplicadas, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México. 2- Laboratorio Nacional de Microscopía Avanzada, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, México. 3- Analytical and Quantitative Light Microscopy, Marine Biological Laboratory, Woods Hole, MA 02543, U.S.A. 4- Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México. Ciudad de México, México. 5- Departamento de Química de Biomacromoléculas, Instituto de Química. Universidad Nacional Autónoma de México. Ciudad de México, Mexico. 6- Instituto de Biología y Medicina Experimental (IBYME‐CONICET), Buenos Aires, Argentina. 7- Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mexico. 8- Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, Mexico. 9- Instituto de Investigaciones en Matemáticas Aplicadas y en Sistemas, Universidad Nacional Autónoma de México, Ciudad de México, Mexico. 10- Department of Biomedicine and Prevention, Faculty of Medicine, University of Rome Tor Vergata, Rome, Italy. 11- Crick Advanced Light Microscopy Facility, United Kingdom. 12- European Molecular Biology Laboratory, Neurobiology and Epigenetics Unit, Monterotondo, Italy. 13- Instituto de Cibernética, Matemática y Física, Ciudad de la Habana, Cuba. 14- Electrical and Computer Engineering and School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, U.S.A.

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.