title: Tunable Single-Cell Extraction for Molecular Analyses.
abstract: Because of cellular heterogeneity, the analysis of endogenous molecules from single cells is of significant interest and has major implications. While micromanipulation or cell sorting followed by cell lysis is already used for subsequent molecular examinations, approaches to directly extract the content of living cells remain a challenging but promising alternative to achieving non-destructive sampling and cell-context preservation. Here, we demonstrate the quantitative extraction from single cells with spatiotemporal control using fluidic force microscopy. We further present a comprehensive analysis of the soluble molecules withdrawn from the cytoplasm or the nucleus, including the detection of enzyme activities and transcript abundances. This approach has uncovered the ability of cells to withstand extraction of up to several picoliters and opens opportunities to study cellular dynamics and cell-cell communication under physiological conditions at the single-cell level.
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich, 8093 Zurich, Switzerland. Electronic address: gorane@ethz.ch.
authors.forename: Orane
authors.initials: O
authors.lastname: Guillaume-Gentil
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Rashel V
authors.initials: RV
authors.lastname: Grindberg
authors.affiliation: Department of Biology, Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Romain
authors.initials: R
authors.lastname: Kooger
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, 8092 Zurich, Switzerland.
authors.forename: Livie
authors.initials: L
authors.lastname: Dorwling-Carter
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, 8092 Zurich, Switzerland.
authors.forename: Vincent
authors.initials: V
authors.lastname: Martinez
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, 8092 Zurich, Switzerland.
authors.forename: Dario
authors.initials: D
authors.lastname: Ossola
authors.affiliation: Department of Biology, Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Pilhofer
authors.affiliation: Laboratory of Biosensors and Bioelectronics, Institute for Biomedical Engineering, ETH Zurich, 8092 Zurich, Switzerland.
authors.forename: Tomaso
authors.initials: T
authors.lastname: Zambelli
authors.affiliation: Department of Biology, Institute of Microbiology, ETH Zurich, 8093 Zurich, Switzerland. Electronic address: jvorholt@ethz.ch.
authors.forename: Julia A
authors.initials: JA
authors.lastname: Vorholt
mesh_terms: D002457:Cell Extracts / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D046529:Microscopy, Electron, Transmission; D036103:Nanotechnology / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods*; D059467:Transcriptome
chemical_list: D002457:Cell Extracts