Light-Sheet Confined Super-Resolution Using Two-Photon Photoactivation |
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| Authors: | Francesca Cella Zanacchi Zeno Lavagnino Mario Faretta Laura Furia Alberto Diaspro |
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| Institution: | 1. Department of Nanophysics, Istituto Italiano di Tecnologia, Genova, Italy.; 2. Università di degli Studi di Genova, Italy.; 3. Department of Experimental Oncology, European Institute of Oncology, Milano, Italy.; University of Zurich, Switzerland, |
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| 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. |
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