title: Live-cell protein labelling with nanometre precision by cell squeezing.
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.
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.
authors.forename: Alina
authors.initials: A
authors.lastname: Kollmannsperger
authors.affiliation: 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.
authors.forename: Armon
authors.initials: A
authors.lastname: Sharei
authors.affiliation: Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue Strasse 7, 60438 Frankfurt/Main, Germany.
authors.forename: Anika
authors.initials: A
authors.lastname: Raulf
authors.affiliation: Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue Strasse 7, 60438 Frankfurt/Main, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Heilemann
authors.affiliation: 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.
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
authors.affiliation: 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.
authors.forename: Klavs F
authors.identifier: http://orcid.org/0000-0001-7192-580X
authors.initials: KF
authors.lastname: Jensen
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.
authors.forename: Ralph
authors.initials: R
authors.lastname: Wieneke
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.|Cluster of Excellence-Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt/Main, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé
mesh_terms: D001696:Biomechanical Phenomena; D002460:Cell Line; D002477:Cells / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D011506:Proteins / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins
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