All-Optical Wide-Field Selective Imaging of Fluorescent Nanodiamonds in Cells, in Vivo and Ex Vivo

Tamami Yanagi, Kiichi Kaminaga, Michiyo Suzuki, Hiroshi Abe, Hiroki Yamamoto, Takeshi Ohshima, Akihiro Kuwahata, Masaki Sekino, Tatsuhiko Imaoka, Shizuko Kakinuma, Takuma Sugi, Wataru Kada, Osamu Hanaizumi, Ryuji Igarashi

研究成果: Article査読

抄録

Fluorescence imaging is a critical tool to understand the spatial distribution of biomacromolecules in cells and in vivo, providing information on molecular dynamics and interactions. Numerous valuable insights into biological systems have been provided by the specific detection of various molecular species. However, molecule-selective detection is often hampered by background fluorescence, such as cell autofluorescence and fluorescence leakage from molecules stained by other dyes. Here we describe a method for all-optical selective imaging of fluorescent nanodiamonds containing nitrogen-vacancy centers (NVCs) for wide-field fluorescence bioimaging. The method is based on the fact that the fluorescence intensity of NVCs strictly depends on the configuration of ground-state electron spins, which can be controlled by changing the pulse recurrence intervals of microsecond excitation laser pulses. Therefore, by using regulated laser pulses, we can oscillate the fluorescence from NVCs in a nanodiamond, while oscillating other optical signals in the opposite phase to NVCs. As a result, we can reconstruct a selective image of a nanodiamond by using a series of oscillated fluorescence images. We demonstrate application of the method to the selective imaging of nanodiamonds in live cells, in microanimals, and on a hippocampal slice culture obtained from a rat. Our approach potentially enables us to achieve high-contrast images of nanodiamond-labeled biomolecules with a signal-to-background ratio improved by up to 100-fold over the standard fluorescence image, thereby providing a more powerful tool for the investigation of molecular dynamics in cells and in vivo.

本文言語English
ページ(範囲)12869-12879
ページ数11
ジャーナルACS Nano
15
8
DOI
出版ステータスPublished - 2021 8 24

ASJC Scopus subject areas

  • 材料科学(全般)
  • 工学(全般)
  • 物理学および天文学(全般)

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