title: Super-resolution two-photon microscopy via scanning patterned illumination.
abstract: We developed two-photon scanning patterned illumination microscopy (2P-SPIM) for super-resolution two-photon imaging. Our approach used a traditional two-photon microscopy setup with temporally modulated excitation to create patterned illumination fields. Combing nine different illuminations and structured illumination reconstruction, super-resolution imaging was achieved in two-photon microscopy. Using 2P-SPIM we achieved a lateral resolution of 141 nm, which represents an improvement by a factor of 1.9 over the corresponding diffraction limit. We further demonstrated super-resolution cellular imaging by 2P-SPIM to image actin cytoskeleton in mammalian cells and three-dimensional imaging in highly scattering retinal tissue.
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
authors.forename: Ben E
authors.initials: BE
authors.lastname: Urban
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
authors.forename: Ji
authors.initials: J
authors.lastname: Yi
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
authors.forename: Siyu
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
authors.forename: Biqin
authors.initials: B
authors.lastname: Dong
authors.affiliation: Department of Ophthalmology, Northwestern University, Chicago, Illinois, USA.
authors.forename: Yongling
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Ophthalmology, Northwestern University, Chicago, Illinois, USA.
authors.forename: Steven H
authors.initials: SH
authors.lastname: DeVries
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.
authors.forename: Vadim
authors.initials: V
authors.lastname: Backman
authors.affiliation: Department of Biomedical Engineering, Northwestern University, Evanston, Illinois, USA.|Department of Ophthalmology, Northwestern University, Chicago, Illinois, USA.
authors.forename: Hao F
authors.initials: HF
authors.lastname: Zhang
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure; D003287:Contrast Media; D004867:Equipment Design; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008962:Models, Theoretical; D054118:Nanospheres / Q000648:ultrastructure; D019047:Phantoms, Imaging; D011806:Quinolinium Compounds; D012160:Retina / Q000648:ultrastructure; D046509:Tissue Culture Techniques
chemical_list: C481052:Alexa fluor 546; D003287:Contrast Media; D011806:Quinolinium Compounds; D049452:Green Fluorescent Proteins