title: PSF decomposition of nanoscopy images via Bayesian analysis unravels distinct molecular organization of the cell membrane.
abstract: The spatial organization of membrane receptors at the nanoscale has major implications in cellular function and signaling. The advent of super-resolution techniques has greatly contributed to our understanding of the cellular membrane. Yet, despite the increased resolution, unbiased quantification of highly dense features, such as molecular aggregates, remains challenging. Here we describe an algorithm based on Bayesian inference of the marker intensity distribution that improves the determination of molecular positions inside dense nanometer-scale molecular aggregates. We tested the performance of the method on synthetic images representing a broad range of experimental conditions, demonstrating its wide applicability. We further applied this approach to STED images of GPI-anchored and model transmembrane proteins expressed in mammalian cells. The analysis revealed subtle differences in the organization of these receptors, emphasizing the role of cortical actin in the compartmentalization of the cell membrane.
authors.affiliation: ICFO-Institut de Ciències Fotòniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona), Spain.
authors.forename: Carlo
authors.initials: C
authors.lastname: Manzo
authors.affiliation: ICFO-Institut de Ciències Fotòniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona), Spain.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: van Zanten
authors.affiliation: National Centre for Biological Sciences (TIFR), Bellary Road, Bangalore 560065, India.
authors.forename: Suvrajit
authors.initials: S
authors.lastname: Saha
authors.affiliation: ICFO-Institut de Ciències Fotòniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona), Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Torreno-Pina
authors.affiliation: National Centre for Biological Sciences (TIFR), Bellary Road, Bangalore 560065, India.
authors.forename: Satyajit
authors.initials: S
authors.lastname: Mayor
authors.affiliation: 1] ICFO-Institut de Ciències Fotòniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona), Spain [2] ICREA-Institució Catalana de Recerca i Estudis Avançats, 08010 Barcelona, Spain.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Garcia-Parajo
mesh_terms: D000199:Actins / Q000737:chemistry / Q000235:genetics; D000465:Algorithms; D000818:Animals; D001499:Bayes Theorem; D016466:CHO Cells; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure*; D003412:Cricetulus; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics; D058975:Folate Receptor 1 / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000295:instrumentation* / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D000099149:Ezrin
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; D058975:Folate Receptor 1; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; D000099149:Ezrin; C052523:actin filament bundling proteins