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Nat Commun. 2016 Jan 29;7:10372. doi: 10.1038/ncomms10372.

Live-cell protein labelling with nanometre precision by cell squeezing.

Author information

1
Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.
2
Department of Chemical Engineering, David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT), 500 Main Street, Building 76-661, Cambridge, Massachusetts 02139, USA.
3
Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue Strasse 7, 60438 Frankfurt/Main, Germany.
4
Cluster of Excellence-Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.

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.

PMID:
26822409
PMCID:
PMC4740111
DOI:
10.1038/ncomms10372
[Indexed for MEDLINE]
Free PMC Article

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