Open AccessLight-Sheet Confined Super-Resolution Using Two-Photon Photoactivation
Author(s) -
Francesca Cella Zanacchi,
Zeno Lavagnino,
Mario Faretta,
Laura Furia,
Alberto Diaspro
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0067667
Subject(s) - light sheet fluorescence microscopy , microscopy , optics , photon , two photon excitation microscopy , resolution (logic) , scattering , light scattering , materials science , excitation , image resolution , physics , computer science , artificial intelligence , fluorescence , scanning confocal electron microscopy , quantum mechanics
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.