A small protein probe for correlated microscopy of endogenous proteinsde 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 journal › Article › Academic › peer-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.
|Number of pages||8|
|Journal||Histochemistry and cell biology|
|Early online date||11-Jan-2018|
|Publication status||Published - Mar-2018|
- Journal Article, ANTIBODIES, TOOLBOX, EPIDERMAL-GROWTH-FACTOR, ELECTRON-MICROSCOPY, FLUORESCENT PROTEIN, QUANTUM DOTS, CELLS, NANOBODIES, LIGHT, VISUALIZATION