Tunable Single-Cell Extraction for Molecular Analyses

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 co...

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Bibliographic Details
Published in:Cell 2016-07, Vol.166 (2), p.506-516
Main Authors: Guillaume-Gentil, Orane, Grindberg, Rashel V., Kooger, Romain, Dorwling-Carter, Livie, Martinez, Vincent, Ossola, Dario, Pilhofer, Martin, Zambelli, Tomaso, Vorholt, Julia A.
Format: Article
Language:English
Subjects:
Cell Extracts - analysis
HeLa Cells
Humans
Microscopy, Atomic Force - methods
Microscopy, Electron, Transmission
Nanotechnology - methods
Single-Cell Analysis - methods
Transcriptome
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Summary: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 We further present a comprehensive analysis of the soluble molecules withdrawn from the cytoplasm or the nucleus, including the detection of enzyme activities and transcript abundances.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. [Display omitted] •Sampling of cytoplasmic and nucleoplasmic fractions from single live cells•Real-time monitoring of the cellular extraction at a sub-picoliter resolution•Extract dispensing strategies adaptable to a broad range of analytical methods•Method allows for ready assessment of enzymatic activities and transcriptional readouts Extraction of sub-picoliter samples of nucleoplasm and cytoplasm from live cells allows cellular heterogeneity to be assessed without killing the cells.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2016.06.025