一种用于在单细胞中追踪受体动力学的实时成像方案。
A live-imaging protocol for tracking receptor dynamics in single cells.
机构信息
Department of Geriatric and General Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
Department of Medicine, University of Toronto, Toronto, ON M5S 1A8, Canada.
出版信息
STAR Protoc. 2022 Apr 22;3(2):101347. doi: 10.1016/j.xpro.2022.101347. eCollection 2022 Jun 17.
Adjacent membrane receptors can show different cellular responses to ligand stimulation. Here, we describe a super-resolution microscopy imaging protocol for tracking the dynamics of two different membrane-bound receptors in single cells. We describe the transfection protocol by electroporation. We detail the imaging procedure for receptors in a single cell. We then outline the data analysis pipeline. We have applied this protocol to imaging of endocytosis of the LOX-1 and AT1 in CHO-K1 cells, but the protocol can be applied to a variety of membrane receptors in other cell lines. For complete details on the use and execution of this protocol, please refer to Takahashi et al. (2021).
相邻的膜受体在受到配体刺激时可能会表现出不同的细胞反应。在这里，我们描述了一种超分辨率显微镜成像方案，用于跟踪单个细胞中两种不同的膜结合受体的动力学。我们描述了通过电穿孔进行转染的方案。我们详细介绍了单个细胞中受体的成像过程。然后，我们概述了数据分析流程。我们已经将该方案应用于 CHO-K1 细胞中 LOX-1 和 AT1 的内吞作用的成像，但该方案可应用于其他细胞系中的各种膜受体。有关该方案使用和执行的完整详细信息，请参阅 Takahashi 等人。（2021 年）。
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