Live-cell protein labelling with nanometre precision by cell squeezing

Nature Communications
Alina KollmannspergerRobert Tampé

Abstract

Live-cell labelling techniques to visualize proteins with minimal disturbance are important; however, the currently available methods are limited in their labelling efficiency, specificity and cell permeability. We describe high-throughput protein labelling facilitated by minimalistic probes delivered to mammalian cells by microfluidic cell squeezing. High-affinity and target-specific tracing of proteins in various subcellular compartments is demonstrated, culminating in photoinduced labelling within live cells. Both the fine-tuned delivery of subnanomolar concentrations and the minimal size of the probe allow for live-cell super-resolution imaging with very low background and nanometre precision. This method is fast in probe delivery (∼ 1,000,000 cells per second), versatile across cell types and can be readily transferred to a multitude of proteins. Moreover, the technique succeeds in combination with well-established methods to gain multiplexed labelling and has demonstrated potential to precisely trace target proteins, in live mammalian cells, by super-resolution microscopy.

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Citations

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Methods Mentioned

BETA
imaging techniques
super-resolution microscopy
flow cytometry
CLIP
super-resolution
fluorescence microscopy
PCR
FCS
transfection
chip

Software Mentioned

FlowJo
Fiji
rapidSTORM
ImageJ

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