Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA.
Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA.
STAR Protoc. 2022 Aug 19;3(3):101611. doi: 10.1016/j.xpro.2022.101611. eCollection 2022 Sep 16.
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).
我们最近开发了一种生物传感器条码化方法，可高度多重跟踪活细胞中的分子活性。在本方案中，我们详细介绍了用一对条码化蛋白对表达不同基因编码荧光生物传感器的细胞进行标记，并进行平行成像。然后将具有相同条码的细胞信号汇集在一起，以获得相应生物传感器活性的动态。我们描述了细胞条码化、图像采集和深度学习模型分析的步骤。如需详细了解本方案的使用和执行，请参考 Yang 等人（2021 年）的文献。
