Temporal focusing microscopy combined with three-dimensional structured illumination

Keisuke Isobe, Keisuke Toda, Qiyuan Song, Fumihiko Kannari, Hiroyuki Kawano, Atsushi Miyawaki, Katsumi Midorikawa

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)


Temporal focusing microscopy provides the optical sectioning capability in wide-field two-photon fluorescence imaging. Here, we demonstrate temporal focusing microscopy combined with three-dimensional structured illumination, which enables us to enhance the three-dimensional spatial resolution and reject the background fluorescence. Experimentally, the periodic pattern of the illumination was produced not only in the lateral direction but also in the axial direction by the interference between three temporal focusing pulses, which were easily generated using a digital micromirror device. The lateral resolution and optical sectioning capability were successfully enhanced by factors of 1.6 and 3.6, respectively, compared with those of temporal focusing microscopy. In the two-photon fluorescence imaging of a tissue-like phantom, the out-of-focus background fluorescence and the scattered background fluorescence could also be rejected.

Original languageEnglish
Article number052501
JournalJapanese journal of applied physics
Issue number5
Publication statusPublished - 2017 May

ASJC Scopus subject areas

  • Engineering(all)
  • Physics and Astronomy(all)


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