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Updated: Apr 3, 2026
Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization
Published on: May 15, 2018
Sample Preparation Methods Following CellSearch Approach Compatible of Single-Cell Whole-Genome Amplification: An
Joost F Swennenhuis1, Leon Terstappen2
1Department 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.
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.
Insights
Isolating rare circulating tumor cells (CTCs) from blood is crucial for understanding cancer. This study reviews five single-cell isolation techniques, highlighting their efficiency, cost, and hands-on time for selecting the best method.
Area of Science:
- Single-cell analysis
- Genomics
- Cancer biology
Background:
- Single cells offer insights into disease heterogeneity and therapy targets.
- Isolating rare cells, like circulating tumor cells (CTCs), presents a significant challenge.
- Existing isolation techniques have limitations and varying capabilities.
Purpose of the Study:
- To evaluate five distinct single-cell isolation techniques.
- To assess these methods for isolating rare circulating tumor cells (CTCs) from pre-enriched blood samples.
- To provide a comparative perspective for selecting optimal isolation platforms.
Main Methods:
- Micromanipulation
- Fluorescence-activated cell sorting (FACS)
- Laser capture microdissection
- DEPArray technology
- Microfluidic solutions
- Evaluation based on CellSearch pre-enriched samples.
Main Results:
- All reviewed platforms exhibit limitations in efficiency.
- Significant differences exist in hands-on time and associated costs among techniques.
- These factors are critical drivers for selecting the most suitable isolation platform.
Conclusions:
- The choice of single-cell isolation technique depends on a balance of efficiency, time, and cost.
- Further optimization of current platforms is needed to improve efficiency for rare cell isolation.
- Effective CTC isolation is vital for advancing cancer diagnostics and treatment strategies.
