Publication

A small protein probe for correlated microscopy of endogenous proteins

de Beer, M. A., Kuipers, J., van Bergen En Henegouwen, P. M. P. & Giepmans, B. N. G., Mar-2018, In : Histochemistry and cell biology. 149, 3, p. 261-268 8 p.

Research output: Contribution to journalArticleAcademicpeer-review

Probes are essential to visualize proteins in their cellular environment, both using light microscopy as well as electron microscopy (EM). Correlated light microscopy and electron microscopy (CLEM) requires probes that can be imaged simultaneously by both optical and electron-dense signals. Existing combinatorial probes often have impaired efficiency, need ectopic expression as a fusion protein, or do not target endogenous proteins. Here, we present FLIPPER-bodies to label endogenous proteins for CLEM. Fluorescent Indicator and Peroxidase for Precipitation with EM Resolution (FLIPPER), the combination of a fluorescent protein and a peroxidase, is fused to a nanobody against a target of interest. The modular nature of these probes allows an easy exchange of components to change its target or color. A general FLIPPER-body targeting GFP highlights histone2B-GFP both in fluorescence and in EM. Similarly, endogenous EGF receptors and HER2 are visualized at nm-scale resolution in ultrastructural context. The small and flexible FLIPPER-body outperforms IgG-based immuno-labeling, likely by better reaching the epitopes. Given the modular domains and possibilities of nanobody generation for other targets, FLIPPER-bodies have high potential to become a universal tool to identify proteins in immuno-CLEM with increased sensitivity compared to current approaches.

Original languageEnglish
Pages (from-to)261-268
Number of pages8
JournalHistochemistry and cell biology
Volume149
Issue number3
Early online date11-Jan-2018
Publication statusPublished - Mar-2018

    Keywords

  • Journal Article, ANTIBODIES, TOOLBOX, EPIDERMAL-GROWTH-FACTOR, ELECTRON-MICROSCOPY, FLUORESCENT PROTEIN, QUANTUM DOTS, CELLS, NANOBODIES, LIGHT, VISUALIZATION

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