Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies

To facilitate an expanded application, researchers present a scalable and high-throughput strategy tosimultaneously target multiple endogenous proteins in living cells with micro- to nanometer resolution. Forintracellular protein labeling, they advanced nanobodies by site-specific and stoichiometric attachment ofbright organicfluorophores. Their fast andfine-tuned intracellular transfer by microfluidic cell squeezingenabled high-throughput delivery with less than 10% dead cells. This strategy allowed for the dual-colorimaging of distinct endogenous cellular structures, and culminated in super-resolution imaging of nativeprotein networks in genetically non-modified living cells. The simultaneous delivery of multipleengineered nanobodies does not only offer exciting prospects for multiplexed imaging of endogenousprotein, but also holds potential for visualizing native cellular structures with unprecedented accuracy.

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