Compartmental Genomics in Living Cells Revealed by Single-Cell Nanobiopsy
| Autor: | Akshar Lohith, Hyunsung John Kim, Michelle M. Maalouf, Paolo Actis, R. Adam Seger, Nader Pourmand, Boaz Vilozny |
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| Rok vydání: | 2013 |
| Předmět: |
nanopipette
Cells 1.1 Normal biological development and functioning Biopsy Cell General Physics and Astronomy Bioengineering Genomics Mitochondrion Biology Polymerase Chain Reaction Genome Article DNA sequencing Single-cell analysis Stem Cell Research - Nonembryonic - Human Underpinning research Genetics medicine Humans Nanotechnology General Materials Science Nanoscience & Nanotechnology Cells Cultured DNA Primers Microscopy Cultured Base Sequence Human Genome General Engineering electrowetting Stem Cell Research Cell biology mitochondria medicine.anatomical_structure single-cell biopsy Scanning ion-conductance microscopy next-generation sequencing Generic health relevance Single-Cell Analysis scanning ion conductance microscopy Intracellular Biotechnology |
| Zdroj: | ACS nano, vol 8, iss 1 |
| ISSN: | 1936-086X 1936-0851 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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