Light-Sheet Confined Super-Resolution Using Two-Photon Photoactivation

Francesca Cella Zanacchi, Zeno Lavagnino, Mario Faretta, Laura Furia, Alberto Diaspro

Research output: Contribution to journalArticle

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.

Original languageEnglish
Article numbere67667
JournalPLoS One
Volume8
Issue number7
DOIs
Publication statusPublished - Jul 2 2013

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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