1. Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy
- Author
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Actis, Paolo, Maalouf, Michelle M, Kim, Hyunsung John, Lohith, Akshar, Vilozny, Boaz, Seger, R Adam, and Pourmand, Nader
- Subjects
Biochemistry and Cell Biology ,Genetics ,Biological Sciences ,Biotechnology ,Human Genome ,Bioengineering ,1.1 Normal biological development and functioning ,Generic health relevance ,Base Sequence ,Biopsy ,Cells ,Cultured ,DNA Primers ,Genomics ,Humans ,Microscopy ,Nanotechnology ,Polymerase Chain Reaction ,Single-Cell Analysis ,nanopipette ,scanning ion conductance microscopy ,single-cell biopsy ,electrowetting ,mitochondria ,next-generation sequencing ,Nanoscience & Nanotechnology - 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.
- Published
- 2014