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.
|Title of host publication||Whole Genome Amplification: Methods and Protocols|
|Publication status||Published - 2015|
|Name||Methods in Molecular Biology|
|Publisher||Springer New York|
Swennenhuis, J. F., & Terstappen, L. W. M. M. (2015). Sample Preparation Methods Following CellSearch Approach Compatible of Single-Cell Whole-Genome Amplification: An Overview. In T. Kroneis (Ed.), Whole Genome Amplification: Methods and Protocols (pp. 57-67). (Methods in Molecular Biology; No. 1347). Springer. https://doi.org/10.1007/978-1-4939-2990-0_4