Astigmatic multifocus microscopy enables deep 3D super-resolved imaging

Laura Oudjedi (1) , Jean-Bernard Fiche (1) , Sara Abrahamsson (2) , Laurent Mazenq (3) , Aurélie Lecestre (3) , Pierre-François Calmon (3) , Aline Cerf (4) , Marcelo Nollmann (1)

1 CBS - Centre de Biochimie Structurale [Montpellier]
2 Rockefeller University [New York]
3 LAAS-TEAM - Service Techniques et Équipements Appliqués à la Microélectronique
4 LAAS-ELIA - Équipe Ingénierie pour les sciences du vivant

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.

Domaines: Physique [physics]

Laura Oudjedi, Jean-Bernard Fiche, Sara Abrahamsson, Laurent Mazenq, Aurélie Lecestre, et al.. Astigmatic multifocus microscopy enables deep 3D super-resolved imaging. Biomedical optics express, 2016, 7 (6), pp.2163. ⟨10.1364/BOE.7.002163⟩. ⟨hal-02060576⟩