STAR Protocols (Jun 2022)

A live-imaging protocol for tracking receptor dynamics in single cells

  • Yibin Huang,
  • Toshimasa Takahashi,
  • Herbert Gaisano,
  • Hiromi Rakugi,
  • Koichi Yamamoto

Journal volume & issue
Vol. 3, no. 2
p. 101347

Abstract

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Summary: 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).

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