Protocol to visualize glycan-mediated binding between virus and human blood mononuclear cells using thin-section transmission electron microscopy

Belinda L Spillings; Kathryn Green; Erica Lovas; Richard Webb; Johnson Mak

doi:10.1016/j.xpro.2023.102673

Protocol to visualize glycan-mediated binding between virus and human blood mononuclear cells using thin-section transmission electron microscopy

STAR Protoc. 2023 Dec 15;4(4):102673. doi: 10.1016/j.xpro.2023.102673. Epub 2023 Oct 27.

Authors

Belinda L Spillings¹, Kathryn Green², Erica Lovas², Richard Webb², Johnson Mak³

Affiliations

¹ Institute for Glycomics, Griffith University, Gold Coast, QLD 4222, Australia. Electronic address: b.devilliers@griffith.edu.au.
² Centre for Microscopy and Microanalysis, The University of Queensland, Brisbane, QLD 4072, Australia.
³ Institute for Glycomics, Griffith University, Gold Coast, QLD 4222, Australia. Electronic address: j.mak@griffith.edu.au.

Abstract

Glycan-glycan interactions between viral particles and host cells may lengthen the dwell time of the virus on the cell surface to facilitate cellular receptor engagement. Here, we present a protocol for visualizing glycan-mediated binding between virus or virus-like-particles (VLPs) and human peripheral blood mononuclear cells using transmission electron microscopy (TEM). We describe steps for virus and VLP production, isolation of human peripheral blood mononuclear cells, and sample preparation. We then detail procedures for thin-section TEM. For complete details on the use and execution of this protocol, please refer to Spillings et al.¹.

Keywords: Cell Biology; Microbiology; Microscopy.

MeSH terms

Cell Line
Cell Membrane
Humans
Leukocytes, Mononuclear*
Microscopy, Electron, Transmission
Virion*

Grants and funding

R21 AI172534/AI/NIAID NIH HHS/United States