title: Light-sheet confined super-resolution using two-photon photoactivation.
abstract: Light-sheet microscopy is a useful tool for performing biological investigations of thick samples and it has recently been demonstrated that it can also act as a suitable architecture for super-resolution imaging of thick biological samples by means of individual molecule localization. However, imaging in depth is still limited since it suffers from a reduction in image quality caused by scattering effects. This paper sets out to investigate the advantages of non-linear photoactivation implemented in a selective plane illumination configuration when imaging scattering samples. In particular, two-photon excitation is proven to improve imaging capabilities in terms of imaging depth and is expected to reduce light-sample interactions and sample photo-damage. Here, two-photon photoactivation is coupled to individual molecule localization methods based on light-sheet illumination (IML-SPIM), allowing super-resolution imaging of nuclear pH2AX in NB4 cells.
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Genova, Italy. francesca.cella@iit.it
authors.forename: Francesca
authors.initials: F
authors.lastname: Cella Zanacchi
authors.forename: Zeno
authors.initials: Z
authors.lastname: Lavagnino
authors.forename: Mario
authors.initials: M
authors.lastname: Faretta
authors.forename: Laura
authors.initials: L
authors.lastname: Furia
authors.forename: Alberto
authors.initials: A
authors.lastname: Diaspro
mesh_terms: D045744:Cell Line, Tumor; D003911:Dextrans; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D006657:Histones / Q000648:ultrastructure; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation / Q000379:methods*; D017785:Photons
chemical_list: D003911:Dextrans; D005456:Fluorescent Dyes; C468783:H2AX protein, human; D006657:Histones; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate