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Luminescent proteins for high-speed single-cell and whole-body imaging

2012; Nature Portfolio; Volume: 3; Issue: 1 Linguagem: Inglês

10.1038/ncomms2248

2041-1723

Kenta Saito, Yu‐Fen Chang, Kazuki Horikawa, Noriyuki Hatsugai, Yuriko Higuchi, Mitsuru Hashida, Yu Yoshida, Tomoki Matsuda, Yoshiyuki Arai, Takeharu Nagai,

Advanced Fluorescence Microscopy Techniques

The use of fluorescent proteins has revolutionized our understanding of biological processes. However, the requirement for external illumination precludes their universal application to the study of biological processes in all tissues. Although light can be created by chemiluminescence, light emission from existing chemiluminescent probes is too weak to use this imaging modality in situations when fluorescence cannot be used. Here we report the development of the brightest luminescent protein to date, Nano-lantern, which is a chimera of enhanced Renilla luciferase and Venus, a fluorescent protein with high bioluminescence resonance energy transfer efficiency. Nano-lantern allows real-time imaging of intracellular structures in living cells with spatial resolution equivalent to fluorescence and sensitive tumour detection in freely moving unshaved mice. We also create functional indicators based on Nano-lantern that can image Ca2+, cyclic adenosine monophosphate and adenosine 5′-triphosphate dynamics in environments where the use of fluorescent indicators is not feasible. These luminescent proteins allow visualization of biological phenomena at previously unseen single-cell, organ and whole-body level in animals and plants. Luminescent proteins are important tools for biomedical imaging but tend to emit fairly little light. Saito et al.. describe a brighter version of a bioluminescent protein that can visualize intracellular dynamics of various signalling molecules with high spatial and temporal resolution.