Multiplexed genetic tags for electron and fluorescence microscopy

Oleksandr Berezin; Alberto Piovesan; Roman Graf; Eleni Samara; Felix Sigmund; Gil Gregor Westmeyer

doi:10.1038/s41596-025-01260-7

Multiplexed genetic tags for electron and fluorescence microscopy

Nat Protoc. 2026 Apr;21(4):1705-1729. doi: 10.1038/s41596-025-01260-7. Epub 2025 Nov 19.

Authors

Oleksandr Berezin^#^{1

2}, Alberto Piovesan^#^{1

2}, Roman Graf^{1

2}, Eleni Samara³, Felix Sigmund^{1

2}, Gil Gregor Westmeyer^{4

5}

Affiliations

¹ Munich Institute of Biomedical Engineering, Department of Bioscience, TUM School of Natural Sciences and TUM School of Medicine, Technical University of Munich, Munich, Germany.
² Institute for Synthetic Biomedicine, Helmholtz Munich, Neuherberg, Germany.
³ Department Circuits-Computation-Models, Max Planck Institute for Biological Intelligence, Martinsried, Germany.
⁴ Munich Institute of Biomedical Engineering, Department of Bioscience, TUM School of Natural Sciences and TUM School of Medicine, Technical University of Munich, Munich, Germany. gil.westmeyer@tum.de.
⁵ Institute for Synthetic Biomedicine, Helmholtz Munich, Neuherberg, Germany. gil.westmeyer@tum.de.

^# Contributed equally.

PMID: 41261217
DOI: 10.1038/s41596-025-01260-7

Abstract

This protocol provides comprehensive guidance for researchers, including those with limited electron microscopy (EM) experience, on the effective use of genetically encoded, multichannel EMcapsulin reporters in both fluorescence microscopy and EM workflows. EMcapsulins are a set of modular gene reporters that manifest as distinct shapes in EM when expressed in cell culture or model organisms. This protocol encompasses detailed instructions for labeling cells or proteins of interest with fluorescent EMcapsulins, along with biochemical quality control measures. Researchers may also adapt the protocol to develop custom EMcapsulin constructs tailored to their specific experimental requirements.

Publication types

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

MeSH terms

Genes, Reporter
Humans
Microscopy, Electron* / methods
Microscopy, Fluorescence / methods

Grants and funding

ERC-COG 865710/EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council)