Saturated-excitation image scanning microscopy

Kenta Temma, Ryosuke Oketani, René Lachmann, Toshiki Kubo, Nicholas I. Smith, Rainer Heintzmann, Katsumasa Fujita

Research output: Contribution to journalArticlepeer-review

Abstract

Image scanning microscopy (ISM) overcomes the trade-off between spatial resolution and signal volume in confocal microscopy by rearranging the signal distribution on a two-dimensional detector array to achieve a spatial resolution close to the theoretical limit achievable by infinitesimal pinhole detection without sacrificing the detected signal intensity. In this paper, we improved the spatial resolution of ISM in three dimensions by exploiting saturated excitation (SAX) of fluorescence. We theoretically investigated the imaging properties of ISM, when the fluorescence signals are nonlinearly induced by SAX, and show combined SAX-ISM fluorescence imaging to demonstrate the improvement of the spatial resolution in three dimensions. In addition, we confirmed that the SNR of SAX-ISM imaging of fluorescent beads and biological samples, which is one of the challenges in conventional SAX microscopy, was improved.

Original languageEnglish
Pages (from-to)13825-13838
Number of pages14
JournalOptics Express
Volume30
Issue number8
DOIs
Publication statusPublished - Apr 11 2022
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics

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