title: Whole Genome Amplification of Labeled Viable Single Cells Suited for Array-Comparative Genomic Hybridization.
abstract: Understanding details of a complex biological system makes it necessary to dismantle it down to its components. Immunostaining techniques allow identification of several distinct cell types thereby giving an inside view of intercellular heterogeneity. Often staining reveals that the most remarkable cells are the rarest. To further characterize the target cells on a molecular level, single cell techniques are necessary. Here, we describe the immunostaining, micromanipulation, and whole genome amplification of single cells for the purpose of genomic characterization. First, we exemplify the preparation of cell suspensions from cultured cells as well as the isolation of peripheral mononucleated cells from blood. The target cell population is then subjected to immunostaining. After cytocentrifugation target cells are isolated by micromanipulation and forwarded to whole genome amplification. For whole genome amplification, we use GenomePlex(®) technology allowing downstream genomic analysis such as array-comparative genomic hybridization.
authors.affiliation: Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21, Graz, 8010, Austria. thomas.kroneis@medunigraz.at.|Sahlgrenska Cancer Center, University of Gothenburg, Gothenburg, Sweden. thomas.kroneis@medunigraz.at.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kroneis
authors.affiliation: Research Unit for Single Cell Analysis, Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21, Graz, 8010, Austria.
authors.forename: Amin
authors.initials: A
authors.lastname: El-Heliebi
mesh_terms: D055028:Comparative Genomic Hybridization / Q000379:methods; D053938:DNA Fragmentation; D015894:Genome, Human; D015698:Genomic Library; D023281:Genomics / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D059010:Single-Cell Analysis / Q000379:methods*
keywords: Immunostaining; Micromanipulation; Single-cell analysis; Whole genome amplification