Saito Kenta, Nakano Masahiro, Nagai Takeharu
The Center for Brain Integration Research, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka, 567-0047, Japan.
Methods Mol Biol. 2016;1461:289-97. doi: 10.1007/978-1-4939-3813-1_24.
Luminescence exerts an ideal optical readout for imaging living subjects including no external light source, whereas the dim luminescence and poor color pallet should be addressed for the better utilities. To address the demerits and to prevail the advantages, we developed a bright luminescent protein, named yellow Nano-lantern, exhibiting about 10-20 times brighter than wild-type RLuc. In this chapter, we demonstrate two luminescence-based protocols in detail: i.e., (a) multicolor visualization of Ca(2+) dynamics in different cellular compartments in a single cell using Ca(2+) indicators based on cyan- and orange-Nano-lanterns and (b) video-rate tumor detection in a freely moving mouse using yellow Nano-lantern.
发光为活体成像提供了理想的光学读出方式,无需外部光源,然而,为了更好地应用,仍需解决发光暗淡和色彩单一的问题。为了克服这些缺点并发挥其优势,我们开发了一种明亮的发光蛋白,名为黄色纳米灯笼,其亮度比野生型海肾荧光素酶高约10至20倍。在本章中,我们详细展示了两种基于发光的方法:即,(a) 使用基于青色和橙色纳米灯笼的钙离子指示剂对单个细胞中不同细胞区室的钙离子动态进行多色可视化,以及(b) 使用黄色纳米灯笼对自由活动小鼠进行视频速率的肿瘤检测。