Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Langer, S; Geigl, JB; Gangnus, R; Speicher, MR.
Sequential application of interphase-FISH and CGH to single cells.
Lab Invest. 2005; 85(4):582-592
Doi: 10.1038/labinvest.3700249
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- Führende Autor*innen der Med Uni Graz
- Speicher Michael
- Co-Autor*innen der Med Uni Graz
- Geigl Jochen Bernd
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- Abstract:
- A comprehensive genomic analysis of single cells is needed for numerous scenarios in tumor genetics, clinical diagnostics and forensic application. PCR protocols were developed which allow an unbiased amplification of the whole genome of a single cell for subsequent analyses by comparative genomic hybridization (CGH). However, verification of single-cell CGH results has been impossible as the procedure naturally involves the destruction of the respective cell. Here we show that the genome of individual cells can be analyzed by two different single cell techniques applied sequentially to the same cell. In a first step, interphase fluorescence in situ hybridization (FISH) is applied. After evaluation of the interphase-FISH signals, cells of interest can be selected for a further analysis. Single cells are collected by laser microdissection, the DNA is amplified by linker-adaptor PCR and subjected to CGH-analysis. This strategy offers new opportunities for a sophisticated selection of cells based on interphase-FISH signals. Furthermore, the sequential application of two different single-cell approaches to the same single-cell represents the only option to control and verify the single-cell CGH results. We demonstrate the feasibility of this approach with a series of experiments including cells from pre- and postnatal diagnostics, for example, cells with trisomies 13, 18, or 21, respectively, leukemia and tumor cells and tissue sections.
- Find related publications in this database (using NLM MeSH Indexing)
- In Situ Hybridization, Fluorescence -
- Interphase -
- Nucleic Acid Hybridization -
- Find related publications in this database (Keywords)
- single-cell analysis
- interphase fluorescence in situ hybridization
- unbiased whole genome amplification
- single-cell comparative genomic hybridization
- tumor heterogeneity