title: PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking.
abstract: We present a novel approach for three-dimensional localization of single molecules using adaptive optics. A 52-actuator deformable mirror is used to both correct aberrations and induce two-dimensional astigmatism in the point-spread-function. The dependence of the z-localization precision on the degree of astigmatism is discussed. We achieve a z-localization precision of 40 nm for fluorescent proteins and 20 nm for fluorescent dyes, over an axial depth of ~800 nm. We illustrate the capabilities of our approach for three-dimensional high-resolution microscopy with super-resolution images of actin filaments in fixed cells and single-molecule tracking of quantum-dot labeled transmembrane proteins in live HeLa cells.
authors.affiliation: Laboratoire Kastler Brossel, CNRS UMR 8552, Département de Physique et Institut de Biologie de l’Ecole Normale Supérieure, Université Pierre et Marie Curie-Paris6, 46 rue d’Ulm 75230 Paris cedex 05, France.
authors.forename: Ignacio
authors.initials: I
authors.lastname: Izeddin
authors.forename: Mohamed
authors.initials: M
authors.lastname: El Beheiry
authors.forename: Jordi
authors.initials: J
authors.lastname: Andilla
authors.forename: Daniel
authors.initials: D
authors.lastname: Ciepielewski
authors.forename: Xavier
authors.initials: X
authors.lastname: Darzacq
authors.forename: Maxime
authors.initials: M
authors.lastname: Dahan
mesh_terms: D004867:Equipment Design; D019544:Equipment Failure Analysis; D006367:HeLa Cells; D006801:Humans; D007089:Image Enhancement / Q000295:instrumentation*; D007909:Lenses; D008853:Microscopy / Q000295:instrumentation*; D011506:Proteins / Q000648:ultrastructure*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D011506:Proteins