%0 Journal Article
%T Multi-experiment nonlinear mixed effect modeling of single-cell translation kinetics after transfection
%A Anita Reiser
%A Daniel Wosch¨¦e
%A Fabian Fr£¿hlich
%A Fabian Joachim Theis
%A Jan Hasenauer
%A Joachim Oskar R£¿dler
%A Laura Fink
%A Thomas Ligon
%J Archive of "NPJ Systems Biology and Applications".
%D 2018
%R 10.1038/s41540-018-0079-7
%X Single-cell translation assay for highly parallel readout of reporter protein expression kinetics after mRNA transfection. a Micropatterned protein arrays are used for highly parallel readout of single-cell kinetics on standardized protein adhesion spots, which enables the observation of thousands of cells over a long time period. The microscopy image shows the micropatterned area of one channel with cells expressing eGFP. b Schematics of the six-channel sample holder and the scanning time-lapse acquisition mode. Stacks of images from individual panels are depicted on the right. c Schematic illustration of the transfection process using mRNA containing lipoplexes. The mRNA, which is released into the cytosol, is translated into a fluorescent reporter protein. The translation dynamics are modeled by biochemical rate equations. d Single-cell eGFP expression is measured by integration over the fluorescence intensity. The zoom-in shows one eGFP-expressing cell confined on a fibronectin square (dashed square). The recording of protein expression begins by adding the mRNA lipoplexes, which are incubated for 1£¿h. e A subset of the single-cell trajectories of eGFP expressing cells shows the heterogeneity within the population. The thick black trajectory corresponds to the mean protein expression dynami
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6288153/