Super-multiplex imaging of cellular dynamics and heterogeneity by integrated stimulated Raman and fluorescence microscopy

Jingwen Shou, Robert Oda, Fanghao Hu, Keiko Karasawa, Mutsuo Nuriya, Masato Yasui, Bruce Shiramizu, Wei Min, Yasuyuki Ozeki

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Observing multiple molecular species simultaneously with high spatiotemporal resolution is crucial for comprehensive understanding of complex, dynamic, and heterogeneous biological systems. The recently reported super-multiplex optical imaging breaks the “color barrier” of fluorescence to achieve multiplexing number over six in living systems, while its temporal resolution is limited to several minutes mainly by slow color tuning. Herein, we report integrated stimulated Raman and fluorescence microscopy with simultaneous multimodal color tunability at high speed, enabling super-multiplex imaging covering diverse molecular contrasts with temporal resolution of seconds. We highlight this technique by demonstrating super-multiplex time-lapse imaging and image-based cytometry of live cells to investigate the dynamics and cellular heterogeneity of eight intracellular components simultaneously. Our technique provides a powerful tool to elucidate spatiotemporal organization and interactions in biological systems.

Original languageEnglish
Article number102832
JournaliScience
Volume24
Issue number8
DOIs
Publication statusPublished - 2021 Aug 20

Keywords

  • Biological sciences research methodologies
  • Cell biology
  • Optics

ASJC Scopus subject areas

  • General

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