UFObow: A single-wavelength excitable Brainbow for simultaneous multicolor ex-vivo and in-vivo imaging of mammalian cells

Author:

Hu Jiahong,Yang Fangfang,Liu Chong,Wang Nengzhi,Xiao Yinghan,Zhai Yujie,Wang Xinru,Zhang Ren,Gao Lulu,Xu Mengli,Wang Jialu,Liu ZhengORCID,Huang Songlin,Liu WenfengORCID,Hu Yajing,Liu Feng,Guo Yuqi,Wang LiangORCID,Yuan JingORCID,Zhang ZhihongORCID,Chu JunORCID

Abstract

AbstractBrainbow is a genetic cell-labeling technique that allows random colorization of multiple cells and real-time visualization of cell fate within a tissue, providing valuable insights into understanding complex biological processes. However, fluorescent proteins (FPs) in Brainbow have distinct excitation spectra with peak difference greater than 35 nm, which requires sequential imaging under multiple excitations and thus leads to long acquisition times. In addition, they are not easily used together with other fluorophores due to severe spectral bleed-through. Here, we report the development of a single-wavelength excitable Brainbow, UFObow, incorporating three newly developed blue-excitable FPs. We have demonstrated that UFObow enables not only tracking the growth dynamics of tumor cells in vivo but also mapping spatial distribution of immune cells within a sub-cubic centimeter tissue, revealing cell heterogeneity. This provides a powerful means to explore complex biology in a simultaneous imaging manner at a single-cell resolution in organs or in vivo.

Publisher

Springer Science and Business Media LLC

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