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Bioluminescent system for dynamic imaging of cell and animal behavior

by: Chikako Hara-Miyauchi, Osahiko Tsuji, Aki Hanyu, Seiji Okada, Akimasa Yasuda, Takashi Fukano, Chihiro Akazawa, Masaya Nakamura, Takeshi Imamura, Yumi Matsuzaki, Hirotaka J. Okano, Atsushi Miyawaki, Hideyuki Okano
Biochemical and Biophysical Research Communications (February 2012), doi:10.1016/j.bbrc.2012.01.141  Key: citeulike:10349143

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Abstract

The current utility of bioluminescence imaging is constrained by a low photon yield that limits temporal sensitivity. Here, we describe an imaging method that uses a chemiluminescent/fluorescent protein, ffLuc-cp156, which consists of a yellow variant of Aequorea GFP and firefly luciferase. We report an improvement in photon yield by over three orders of magnitude over current bioluminescent systems. We imaged cellular movement at high resolution including neuronal growth cones and microglial cell protrusions. Transgenic ffLuc-cp156 mice enabled video-rate bioluminescence imaging of freely moving animals, which may provide a reliable assay for drug distribution in behaving animals for pre-clinical studies. ⺠We combined a yellow variant of GFP and firefly luciferase to make ffLuc-cp156. ⺠ffLuc-cp156 showed improved photon yield in cultured cells and transgenic mice. ⺠ffLuc-cp156 enabled video-rate bioluminescence imaging of freely-moving animals. ⺠ffLuc-cp156 mice enabled tracking real-time drug delivery in conscious animals.


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