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

Live-cell protein labelling with nanometre precision by cell squeezing

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Author(s)
Kollmannsperger, Alina
•
Raulf, Anika
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Heilemann, Mike
•
Wieneke, Ralph
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Tampe, Robert
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Sharei, Armon Reza
•
Langer, Robert S
•
Jensen, Klavs F
Date Issued
January 2016
Journal
Nature Communications
Publisher
Nature Publishing Group
Citation
Kollmannsperger, Alina, Armon Sharei, Anika Raulf, Mike Heilemann, Robert Langer, Klavs F. Jensen, Ralph Wieneke, and Robert Tampe. “Live-Cell Protein Labelling with Nanometre Precision by Cell Squeezing.” Nat Comms 7 (January 29, 2016): 10372.
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Final published version
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution
http://creativecommons.org/licenses/by/4.0/
Persistent DSpace Link
http://hdl.handle.net/1721.1/101097
DOI of Published Version
https://doi.org/10.1038/ncomms10372
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