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Live-cell protein labelling with nanometre precision by cell squeezing

Author

Listed:
  • 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
    Cluster of Excellence—Macromolecular Complexes, Goethe-University Frankfurt)

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.

Suggested Citation

  • Alina Kollmannsperger & Armon Sharei & Anika Raulf & Mike Heilemann & Robert Langer & Klavs F. Jensen & Ralph Wieneke & Robert Tampé, 2016. "Live-cell protein labelling with nanometre precision by cell squeezing," Nature Communications, Nature, vol. 7(1), pages 1-7, April.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms10372
    DOI: 10.1038/ncomms10372
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