@article{3dd1885c294f439ea238d45e8b9f6b58,
title = "Imaging and analysis for simultaneous tracking of fluorescent biosensors in barcoded cells",
abstract = "We recently developed a biosensor barcoding approach for highly multiplexed tracking of molecular activities in live cells. In this protocol, we detail the labeling of cells expressing different genetically encoded fluorescent biosensors with a pair of barcoding proteins and parallel imaging. Signals from cells with the same barcodes are then pooled together to obtain the dynamics of the corresponding biosensor activity. We describe the steps involved in cell barcoding, image acquisition, and analysis by deep learning models. For complete details on the use and execution of this protocol, please refer to Yang et al. (2021).",
keywords = "Cell biology, Microscopy, Molecular/Chemical probes, Signal transduction, Single cell",
author = "Chi, {Wei Yu} and Gabriel Au and Jessica Liang and Chen, {Chao Cheng} and Huang, {Chuan Hsiang} and Yang, {Jr Ming}",
note = "Funding Information: The authors would like to thank Michiyuki Matsuda for the plasmids of PicchuEV, EV-ROCK, and EV-S6K. This work was supported by NIH grants K22CA212060 (to C.H.H.), R01GM136711 (to C.H.H.), Cervical Cancer SPORE P50CA098252 Career Development Award (to J.M.Y.) and Pilot Project Award (to C.H.H.), the W. W. Smith Charitable Trust (#C1901, to C.H.H.) and the Sol Goldman Pancreatic Cancer Research Center (to C.H.H.). The Zeiss LSM 780 confocal microscope was purchased with NIH grants S10OD016374. W.Y.C. G.A. C.H.H. and J.M.Y. wrote the manuscript with inputs from J.L. and C.C.C. C.H.H. and J.M.Y. supervised the project. All authors read and approved the manuscript. The authors declare no competing interests. Funding Information: The authors would like to thank Michiyuki Matsuda for the plasmids of PicchuEV, EV-ROCK, and EV-S6K. This work was supported by NIH grants K22CA212060 (to C.H.H.), R01GM136711 (to C.H.H.), Cervical Cancer SPORE P50CA098252 Career Development Award (to J.M.Y.) and Pilot Project Award (to C.H.H.), the W. W. Smith Charitable Trust (# C1901 , to C.H.H.) and the Sol Goldman Pancreatic Cancer Research Center (to C.H.H.). The Zeiss LSM 780 confocal microscope was purchased with NIH grants S10OD016374 . Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = sep,
day = "16",
doi = "10.1016/j.xpro.2022.101611",
language = "English (US)",
volume = "3",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "3",
}