title: Sample Preparation Methods Following CellSearch Approach Compatible of Single-Cell Whole-Genome Amplification: An Overview.
abstract: Single cells are increasingly used to determine the heterogeneity of therapy targets in the genome during the course of a disease. The first challenge using single cells is to isolate these cells from the surrounding cells, especially when the targeted cells are rare. A number of techniques have been developed for this goal, each having specific limitations and possibilities. In this chapter, five of these techniques are discussed in the light of the isolation of circulating tumor cells (CTC) present at extremely low frequency in the blood of patients with metastatic cancer from the perspective of pre-enriched samples by means of CellSearch. The techniques described are micromanipulation, FACS, laser capture microdissection, DEPArray, and microfluidic solutions. All platforms are hampered with a low efficiency and differences in hands-on time and costs are the most important drivers for selection of the optimal platform.
authors.affiliation: Department of Medical Cell BioPhysics, Faculty of Science and Technology, MIRA Research Institute, University of Twente, 217, Drienerlolaan 5, 7500 AE, Enschede, The Netherlands. j.f.swennenhuis@utwente.nl.
authors.forename: Joost F
authors.initials: JF
authors.lastname: Swennenhuis
authors.affiliation: Department of Medical Cell BioPhysics, Faculty of Science and Technology, MIRA Research Institute, University of Twente, 217, Drienerlolaan 5, 7500 AE, Enschede, The Netherlands.
authors.forename: Leon
authors.initials: L
authors.lastname: Terstappen
mesh_terms: D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods; D016678:Genome; D023281:Genomics / Q000379:methods*; D006801:Humans; D061346:Laser Capture Microdissection / Q000379:methods; D044085:Microfluidics / Q000379:methods; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology; D021141:Nucleic Acid Amplification Techniques; D059010:Single-Cell Analysis / Q000379:methods*
keywords: CellSearch; Circulating tumor cells; DEPArray; Laser capture microdissection; Microfluidics; Micromanipulation; Rare cells; Sample pre-enrichment; Single-cell whole-genome amplification