Abstract
The handling of single cells is of great importance in applications such as cell line development or single-cell analysis, e.g., for cancer research or for emerging diagnostic methods. This review provides an overview of technologies that are currently used or in development to isolate single cells for subsequent single-cell analysis. Data from a dedicated online market survey conducted to identify the most relevant technologies, presented here for the first time, shows that FACS (fluorescence activated cell sorting) respectively Flow cytometry (33% usage), laser microdissection (17%), manual cell picking (17%), random seeding/dilution (15%), and microfluidics/lab-on-a-chip devices (12%) are currently the most frequently used technologies. These most prominent technologies are described in detail and key performance factors are discussed. The survey data indicates a further increasing interest in single-cell isolation tools for the coming years. Additionally, a worldwide patent search was performed to screen for emerging technologies that might become relevant in the future. In total 179 patents were found, out of which 25 were evaluated by screening the title and abstract to be relevant to the field.
Highlights
With regards to heterogeneous cell populations, such as those found in many tumors, the generation and analysis of single cells has an increasing impact on various fields of life sciences and biomedical research [1]
To increase the flexibility and enable easier interfacing with other upstream and downstream methods, the authors of this review have proposed previously a flexible single-cell isolation system (Figure 6), based on inkjet-like single-cell printing [52,53]
Regarding performance fluorescence activated cell sorters (FACS) systems have the main benefit of high throughput and sorting capability but are cost-intensive and potentially harmful to some cells
Summary
With regards to heterogeneous cell populations, such as those found in many tumors, the generation and analysis of single cells has an increasing impact on various fields of life sciences and biomedical research [1]. A further need for the analysis of single-cells relates to very rare cells like circulating tumor cells, which are surrounded by billions of normal blood cells and have an increasing clinical impact as a so-called liquid biopsy [4]. At present, the isolation and separation of single cells is still a technically challenging task. Main challenges are the yield and quality or in other words the integrity and purity of the cells as well as the throughput and the sensitivity of single cell isolation methods. A large variety of technologies for single-cell separation, isolation, and sorting are already available that are applied according to the scientific objective
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