Imaging and analysis for simultaneous tracking of fluorescent biosensors in barcoded cells

Summary: 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 imagin...

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Bibliographic Details
Main Authors: Wei-Yu Chi, Gabriel Au, Jessica Liang, Chao-Cheng Chen, Chuan-Hsiang Huang, Jr-Ming Yang
Format: Article
Language:English
Published: Elsevier 2022-09-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166722004919
Description
Summary:Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667