Fan Jin’s team provided a complete solution for simultaneous use of multicolor fluorescent proteins, including a molecular biology toolbox for multicolor fluorescent systems and precise separation algorithms for multiple fluorescent signals. The tool kit contains a promoter module, a fluorescent protein module and a vector module, which can realize one-step assembly and construction of 1-4 color fluorescent clone plasmids. The separation algorithm solves the concentration of each fluorescent protein in the microbial cell through a linear solution equation system, and uses the protein concentration information to characterize the expression level, which can realize the precise separation of specific signals from the mixed fluorescent signals contributed by different fluorescent proteins. Through the above scheme, we can quickly and easily construct multicolor fluorescent plasmids. It is important that the protein concentration information obtained by algorithm analysis is independent of each other and does not depend on the shooting parameters and equipment, and can compare data between different equipment. This solution not only provides a technical method for multicolor fluorescence imaging, but also the application of this method will greatly promote our related research on the relationship between genes in natural or synthetic gene networks.
The achievement was recently published in the international academic journal ACS Synthetic Biology titled "Simultaneous Visualization of Multiple Gene Expression in Single Cells Using an Engineered Multicolor Reporter Toolbox and Approach of Spectral Crosstalk Correction" (10.1021 / acssynbio.9b00223). This research was supported by the National Natural Science Foundation of China and the basic research funding of central universities.
(Left) Schematic Diagram of Plasmid System for 1-4 Color Fluorescence Reporting System; (right) Five-color Fluorescence Imaging After Cross-color Correction.
Article link:https://pubs.acs.org/doi/abs/10.1021/acssynbio.9b00223