Selective-plane-activation structured illumination microscopy

Kenta Temma; Ryosuke Oketani; Toshiki Kubo; Kazuki Bando; Shunsuke Maeda; Kazunori Sugiura; Tomoki Matsuda; Rainer Heintzmann; Tatsuya Kaminishi; Koki Fukuda; Maho Hamasaki; Takeharu Nagai; Katsumasa Fujita

doi:10.1038/s41592-024-02236-3

Selective-plane-activation structured illumination microscopy

Nat Methods. 2024 Apr 5. doi: 10.1038/s41592-024-02236-3. Online ahead of print.

Authors

Kenta Temma^#^{1

2

3}, Ryosuke Oketani^#^{1

4}, Toshiki Kubo¹, Kazuki Bando¹, Shunsuke Maeda¹, Kazunori Sugiura⁵, Tomoki Matsuda⁵, Rainer Heintzmann^{6

7}, Tatsuya Kaminishi^{3

8}, Koki Fukuda^{3

9}, Maho Hamasaki^{8

9}, Takeharu Nagai^{3

5

10}, Katsumasa Fujita^{11

12

13}

Affiliations

¹ Department of Applied Physics, Osaka University, Osaka, Japan.
² Advanced Photonics and Biosensing Open Innovation Laboratory, AIST-Osaka University, Osaka, Japan.
³ Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Osaka, Japan.
⁴ Department of Chemistry, Kyushu University, Fukuoka, Japan.
⁵ SANKEN (The Institute of Scientific and Industrial Research), Osaka University, Osaka, Japan.
⁶ Leibniz Institute of Photonic Technology, Jena, Germany.
⁷ Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller University Jena, Jena, Germany.
⁸ Department of Genetics, Graduate School of Medicine, Osaka University, Osaka, Japan.
⁹ Laboratory of Intracellular Membrane Dynamics, Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
¹⁰ Research Institute for Electronic Science, Hokkaido University, Hokkaido, Japan.
¹¹ Department of Applied Physics, Osaka University, Osaka, Japan. fujita@ap.eng.osaka-u.ac.jp.
¹² Advanced Photonics and Biosensing Open Innovation Laboratory, AIST-Osaka University, Osaka, Japan. fujita@ap.eng.osaka-u.ac.jp.
¹³ Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Osaka, Japan. fujita@ap.eng.osaka-u.ac.jp.

^# Contributed equally.

PMID: 38580844
DOI: 10.1038/s41592-024-02236-3

Abstract

The background light from out-of-focus planes hinders resolution enhancement in structured illumination microscopy when observing volumetric samples. Here we used selective plane illumination and reversibly photoswitchable fluorescent proteins to realize structured illumination within the focal plane and eliminate the out-of-focus background. Theoretical investigation of the imaging properties and experimental demonstrations show that selective plane activation is beneficial for imaging dense microstructures in cells and cell spheroids.

Abstract

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