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Review
. 2015 Jul 24;16(8):16897-919.
doi: 10.3390/ijms160816897.

Technologies for Single-Cell Isolation

Andre Gross  1   2 , Jonas Schoendube  3   4 , Stefan Zimmermann  5 , Maximilian Steeb  6 , Roland Zengerle  7   8   9 , Peter Koltay  10   11
Affiliations
Review

Technologies for Single-Cell Isolation

Andre Gross et al. Int J Mol Sci. .

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.

Keywords: flow cytometry; laser microdissection; limiting dilution; microfluidics; single-cell analysis; single-cell handling; single-cell separation; single-cell technologies.

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Figures

Figure 1
Figure 1
(a) Distribution of participants in their general fields of research. 44 participants stated basic research and 30 applied research as their general field; and (b) Distribution of participants in their specific fields of research. Immunology and oncology were most frequently named.
Figure 2
Figure 2
(a) The usage of technologies for handling single-cells in Germany in 2014. This data was derived as part of this work by a survey amongst 210 participants from German universities, research institutes and industry; and (b) Extensiveness of use of different single-cell technologies (data from “Single Cell Technologies Trends 2014” [6], reproduced with permission from HTStec Limited, Single Cell Technologies Trends 2014, HTStec 2014 URL: http://selectbiosciences.com/ MarketReportsID.aspx?reportID=83).
Figure 3
Figure 3
Schematic overview of single-cell separation technologies discussed in the following. The five technologies were identified through market studies as the most commonly used technologies for the handling of single cells (cf. (compare to) Figure 1).
Figure 4
Figure 4
Schematic view on laser capture microdissection (LCM) methods. (a) Contact-based via adhesive tapes; (b) Cutting with a focused laser followed by capture with a vessel. Cut-out section extracted by gravity; and (c) Cutting with a focused laser followed by pressure catapulting with a defocused laser pulse.
Figure 5
Figure 5
Schematic overview of different microfluidic methods for single-cell isolation. (a) An aqueous stream of cells is broken up into individual droplets-in-oil containing random distribution of cells; (b) Pneumatic membrane valves use air pressure to close a microfluidic channel by membrane deflection. This stops the flow and can trap a cell; and (c) Hydrodynamic traps are passive elements that only fit single cells and hold them at one position.
Figure 6
Figure 6
Single-cell printer (SCP) for single-cell isolation. A microfluidic dispenser chip integrated in a polymer cartridge is filled with cell suspension. An automated object recognition algorithm detects cells in the dispenser nozzle prior to the dispensation. This allows for ejection of droplets containing one single-cell only and their deposition in direction of the arrow on various substrates, such as micro-well plates.
Figure 7
Figure 7
Importance of single-cell analysis to German cell researchers in 2014 and estimated for 2017. This data was derived by a survey amongst 210 participants from German universities, research institutes and industry. A strong growth of interest is expected over the next years.
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