CUED Publications database

Graphene Liquid Enclosure for Single-Molecule Analysis of Membrane Proteins in Whole Cells Using Electron Microscopy

Dahmke, IN and Verch, A and Hermannsdörfer, J and Peckys, DB and Weatherup, RS and Hofmann, S and De Jonge, N (2017) Graphene Liquid Enclosure for Single-Molecule Analysis of Membrane Proteins in Whole Cells Using Electron Microscopy. ACS Nano, 11. pp. 11108-11117. ISSN 1936-0851

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Abstract

Membrane proteins govern many important functions in cells via dynamic oligomerization into active complexes. However, analytical methods to study their distribution and functional state in relation to the cellular structure are currently limited. Here, we introduce a technique for studying single-membrane proteins within their native context of the intact plasma membrane. SKBR3 breast cancer cells were grown on silicon microchips with thin silicon nitride windows. The cells were fixed, and the epidermal growth factor receptor ErbB2 was specifically labeled with quantum dot (QD) nanoparticles. For correlative fluorescence- and liquid-phase electron microscopy, we enclosed the liquid samples by chemical vapor deposited (CVD) graphene films. Depending on the local cell thickness, QD labels were imaged with a spatial resolution of 2 nm at a low electron dose. The distribution and stoichiometric assembly of ErbB2 receptors were determined at several different cellular locations, including tunneling nanotubes, where we found higher levels of homodimerization at the connecting sites. This experimental approach is applicable to a wide range of cell lines and membrane proteins and particularly suitable for studies involving both inter- and intracellular heterogeneity in protein distribution and expression.

Item Type: Article
Uncontrolled Keywords: STEM breast cancer cell epidermal growth factor receptor graphene liquid-phase electron microscopy single-molecule analysis tunneling nanotube Cell Line, Tumor Graphite Humans Lab-On-A-Chip Devices Membrane Proteins Microscopy, Electron Neoplasm Proteins Quantum Dots Receptor, ErbB-2 Silicon Compounds Single Molecule Imaging
Subjects: UNSPECIFIED
Divisions: Div B > Solid State Electronics and Nanoscale Science
Depositing User: Cron Job
Date Deposited: 25 Oct 2017 20:05
Last Modified: 10 Apr 2021 22:19
DOI: 10.1021/acsnano.7b05258