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

Yibin Huang; Toshimasa Takahashi; Herbert Gaisano; Hiromi Rakugi; Koichi Yamamoto

doi:10.1016/j.xpro.2022.101347

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

STAR Protoc. 2022 Apr 22;3(2):101347. doi: 10.1016/j.xpro.2022.101347. eCollection 2022 Jun 17.

Authors

Yibin Huang¹, Toshimasa Takahashi¹, Herbert Gaisano², Hiromi Rakugi¹, Koichi Yamamoto¹

Affiliations

¹ Department of Geriatric and General Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
² Department of Medicine, University of Toronto, Toronto, ON M5S 1A8, Canada.

Abstract

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).

Keywords: Cell Biology; Cell culture; Microscopy; Signal Transduction; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Endocytosis*
Microscopy* / methods