title: Compartmental genomics in living cells revealed by single-cell nanobiopsy.
abstract: The ability to study the molecular biology of living single cells in heterogeneous cell populations is essential for next generation analysis of cellular circuitry and function. Here, we developed a single-cell nanobiopsy platform based on scanning ion conductance microscopy (SICM) for continuous sampling of intracellular content from individual cells. The nanobiopsy platform uses electrowetting within a nanopipette to extract cellular material from living cells with minimal disruption of the cellular milieu. We demonstrate the subcellular resolution of the nanobiopsy platform by isolating small subpopulations of mitochondria from single living cells, and quantify mutant mitochondrial genomes in those single cells with high throughput sequencing technology. These findings may provide the foundation for dynamic subcellular genomic analysis.
authors.affiliation: Department of Biomolecular Engineering, University of California Santa Cruz , 1156 High Street, Santa Cruz, California 95064, United States.
authors.forename: Paolo
authors.initials: P
authors.lastname: Actis
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Maalouf
authors.forename: Hyunsung John
authors.initials: HJ
authors.lastname: Kim
authors.forename: Akshar
authors.initials: A
authors.lastname: Lohith
authors.forename: Boaz
authors.initials: B
authors.lastname: Vilozny
authors.forename: R Adam
authors.initials: RA
authors.lastname: Seger
authors.forename: Nader
authors.initials: N
authors.lastname: Pourmand
mesh_terms: D001483:Base Sequence; D001706:Biopsy / Q000379:methods*; D002478:Cells, Cultured; D017931:DNA Primers; D023281:Genomics; D006801:Humans; D008853:Microscopy / Q000379:methods; D036103:Nanotechnology; D016133:Polymerase Chain Reaction; D059010:Single-Cell Analysis
chemical_list: D017931:DNA Primers