Cryogenic measurement of photophysical properties of a ratiometric fluorescent calcium-ion indicator, Yellow Cameleon 3.60

Masahito Yamanaka; Wakana Miyamura; Takumi Kunimoto; Toshiki Kubo; Kosuke Tsuji; Kazunori Sugiura; Shu-ichi Fukushima; Nicholas I. Smith; Takeharu Nagai; Katsumasa Fujita

doi:10.1117/12.3002051

13 March 2024 Cryogenic measurement of photophysical properties of a ratiometric fluorescent calcium-ion indicator, Yellow Cameleon 3.60

Masahito Yamanaka, Wakana Miyamura, Takumi Kunimoto, Toshiki Kubo, Kosuke Tsuji, Kazunori Sugiura, Shu-ichi Fukushima, Nicholas I. Smith, Takeharu Nagai, Katsumasa Fujita

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Proceedings Volume PC12862, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications XV; PC128620H (2024) https://doi.org/10.1117/12.3002051
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

We investigated the photophysical property of Yellow Cameleon 3.60 (YC3.60), a fluorescent calcium-ion (Ca2+) indicator based on Förster resonance energy transfer (FRET), under cryogenic conditions. By measuring the fluorescence intensity ratio of the donor and accepter at various Ca2+ concentrations under room and cryogenic temperatures, we confirmed that YC3.60 exhibits a Ca2+-dependent FRET efficiency. Although slight differences were observed in the fluorescence lifetime and spectral shape at the cryogenic temperature, which can affect the FRET efficiency, our measurement suggested that YC3.60 can be employed for quantitative Ca2+ measurement and imaging under cryogenic conditions with improved photostability and quantum yield.

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Masahito Yamanaka, Wakana Miyamura, Takumi Kunimoto, Toshiki Kubo, Kosuke Tsuji, Kazunori Sugiura, Shu-ichi Fukushima, Nicholas I. Smith, Takeharu Nagai, and Katsumasa Fujita "Cryogenic measurement of photophysical properties of a ratiometric fluorescent calcium-ion indicator, Yellow Cameleon 3.60", Proc. SPIE PC12862, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications XV, PC128620H (13 March 2024); https://doi.org/10.1117/12.3002051

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KEYWORDS

Cryogenics

Fluorescence

Fluorescence resonance energy transfer

Venus

Fluorescent proteins

Quantum efficiency

Quantum reading

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