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Near-Infrared Bioluminescence Imaging of Animal Cells with Through-Bond Energy Transfer Cassette

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Live Cell Imaging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2274))

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Abstract

Coelenterazine (CTZ) is the most general substrate for marine luciferases. The present protocol introduces a near-infrared (NIR ) bioluminescence (BL) imaging of mammalian cells with a cyanine-5 (Cy5) dye-conjugated CTZ . This unique Cy5-conjugated CTZ, named Cy5-CTZ , can act as a dual optical readout emitting both fluorescence (FL) and BL. The Cy5-CTZ exerts through-bond energy transfer (TBET)-based imaging modalities for mammalian cells. This novel derivative, Cy5-CTZ , is intrinsically fluorescent and emits NIR-shifted BL when reacting with an appropriate luciferase , such as Renilla luciferase (RLuc). The protocol exemplifies a unique live-cell imaging with Cy5-CTZ that is optically stable in physiological samples and rapidly permeabilize through plasma membrane and emit NIR-BL in live mammalian cells.

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Acknowledgments

This work was partly supported by JSPS KAKENHI Grants: Numbers 26288088, 15KK0029, 17H01215, 21H04948, and 24225001.

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Authors and Affiliations

  1. Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama, Japan

    Masahiro Abe, Ryo Nishihara & Koji Suzuki

  2. Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan

    Sung-Bae Kim

Authors
  1. Masahiro Abe
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  2. Ryo Nishihara
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  3. Sung-Bae Kim
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  4. Koji Suzuki
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Corresponding author

Correspondence to Sung-Bae Kim .

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Editors and Affiliations

  1. National Institute of Advanced Industrial Science and Technology, Research Institute for Environmental Management Technology, Tsukuba, Japan

    Sung-Bae Kim

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Abe, M., Nishihara, R., Kim, SB., Suzuki, K. (2021). Near-Infrared Bioluminescence Imaging of Animal Cells with Through-Bond Energy Transfer Cassette. In: Kim, SB. (eds) Live Cell Imaging. Methods in Molecular Biology, vol 2274. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1258-3_10

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  • DOI: https://doi.org/10.1007/978-1-0716-1258-3_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1257-6

  • Online ISBN: 978-1-0716-1258-3

  • eBook Packages: Springer Protocols

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