Super-Resolution Live Imaging of Cargo Traffic Through the Golgi Apparatus in Mammalian Cells

Takuro Tojima; Daisuke Miyashiro; Yasuhito Kosugi; Akihiko Nakano

doi:10.1007/978-1-0716-2639-9_10

Super-Resolution Live Imaging of Cargo Traffic Through the Golgi Apparatus in Mammalian Cells

Methods Mol Biol. 2023:2557:127-140. doi: 10.1007/978-1-0716-2639-9_10.

Authors

Takuro Tojima¹, Daisuke Miyashiro², Yasuhito Kosugi, Akihiko Nakano²

Affiliations

¹ Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Wako, Saitama, Japan. takuro.tojima@riken.jp.
² Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Wako, Saitama, Japan.

PMID: 36512214
DOI: 10.1007/978-1-0716-2639-9_10

Abstract

Super-resolution confocal live imaging microscopy (SCLIM) we developed provides high-speed, high-resolution, three- and four-dimensional, and multicolor simultaneous imaging. Using this technology, we are now able to observe the fine details of various dynamic events going on in living cells, such as membrane traffic and organelle dynamics. The retention using selective hooks (RUSH) system is a powerful tool to control synchronous release of natural cargo proteins of interest from the endoplasmic reticulum in mammalian cells. In this chapter, we describe a method for visualizing secretory cargo traffic within and around the Golgi apparatus in HeLa cells using SCLIM in combination with the RUSH assay.

Keywords: Golgi apparatus; Live cell imaging; Membrane traffic; Retention using selective hooks (RUSH); Super-resolution microscopy.

Publication types

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

MeSH terms

Animals
Endoplasmic Reticulum* / metabolism
Golgi Apparatus* / metabolism
HeLa Cells
Humans
Mammals
Microscopy, Confocal / methods
Protein Transport