title: Astigmatic multifocus microscopy enables deep 3D super-resolved imaging.
abstract: We have developed a 3D super-resolution microscopy method that enables deep imaging in cells. This technique relies on the effective combination of multifocus microscopy and astigmatic 3D single-molecule localization microscopy. We describe the optical system and the fabrication process of its key element, the multifocus grating. Then, two strategies for localizing emitters with our imaging method are presented and compared with a previously described deep 3D localization algorithm. Finally, we demonstrate the performance of the method by imaging the nuclear envelope of eukaryotic cells reaching a depth of field of ~4µm.
authors.affiliation: Centre de Biochimie Structurale, CNRS UMR5048, INSERM U1054, Université de Montpellier, 29 Rue de Navacelles, 34090 Montpellier, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Oudjedi
authors.affiliation: Centre de Biochimie Structurale, CNRS UMR5048, INSERM U1054, Université de Montpellier, 29 Rue de Navacelles, 34090 Montpellier, France.
authors.forename: Jean-Bernard
authors.initials: JB
authors.lastname: Fiche
authors.affiliation: Lulu and Anthony Wang Laboratory of Neural Circuits and Behavior, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Sara
authors.initials: S
authors.lastname: Abrahamsson
authors.affiliation: CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France; Université de Toulouse, LAAS, F-31031 Toulouse, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Mazenq
authors.affiliation: CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France; Université de Toulouse, LAAS, F-31031 Toulouse, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Lecestre
authors.affiliation: CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France; Université de Toulouse, LAAS, F-31031 Toulouse, France.
authors.forename: Pierre-François
authors.initials: PF
authors.lastname: Calmon
authors.affiliation: CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France; Université de Toulouse, LAAS, F-31031 Toulouse, France.
authors.forename: Aline
authors.initials: A
authors.lastname: Cerf
authors.affiliation: Centre de Biochimie Structurale, CNRS UMR5048, INSERM U1054, Université de Montpellier, 29 Rue de Navacelles, 34090 Montpellier, France.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Nöllmann
keywords: (050.1380) Binary optics; (050.1950) Diffraction gratings; (100.6640) Superresolution; (110.1080) Active or adaptive optics; (180.2520) Fluorescence microscopy; (180.6900) Three-dimensional microscopy