Live-cell protein labelling with nanometre precision by cell squeezing

Alina Kollmannsperger, Armon Sharei, Anika Raulf, Mike Heilemann, Robert Langer, Klavs F. Jensen, Ralph Wieneke () and Robert Tampé ()
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Alina Kollmannsperger: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt
Armon Sharei: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT)
Anika Raulf: Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt
Mike Heilemann: Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt
Robert Langer: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT)
Klavs F. Jensen: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT)
Ralph Wieneke: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt
Robert Tampé: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt

Nature Communications, 2016, vol. 7, issue 1, 1-7

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms10372

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