Technologies for Single-Cell Isolation

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.

Microchips and their Significance in Isolation of Circulating Tumor Cells and Monitoring of Cancers

“…One of the main challenges encounter with the cell isolation is the heterogeneous nature of target cells which highlights the needs for the development of single cell isolation and analysis techniques (Van Loo and Voet, 2014). With the respective of frequency, the most applicable technologies which currently used for single cell isolation purposes are flow cytometry (33%), manual cell picking (17%), laser micro-dissection (17%), random seeding/dilution (15%), and microchips (12%) (Gross et al, 2015). Among these, laser-directed micro-dissection is a strong method for the collection of malignant cells in solid tumors (Emmert-Buck et al, 1996).…”

Single-Cell Metabolomics by Mass Spectrometry for Drug Discovery: Moving Forward

“…Additionally, a low-calcium medium may reduce undesirable release of analytes from neurons. Current methods for single cell isolation include serial dilution, micromanipulation, laser capture micro-dissection, fluorescence-activated cell sorting (FACS), microfluidics, manual picking and Raman tweezers (Figure 2) [8]. …”

Design and Fabrication of a Dielectrophoretic Cell Trap Array

“…Briefly, in MACS, the target cells are labeled with magnetic beads and they are separated from the cell mixture by applying a magnetic force [5]. Therefore, the prior knowledge about the surface proteins of the cells in the mixture is required for both FACS and MACS methods [3]. Unfortunately, the information of target cells' surface proteins is not always available and therefore, both of these methods can only be used to isolate limited number of target cells from cell mixtures.…”

“…FACS is the most commonly used method but, to use FACS to isolate target cells, complete knowledge about the surface proteins of the target cells and other non-target cells in the cell mixture is needed. In FACS, first, a fluorescence dye is selectively attached to a surface protein of the target cell and then the separation of target cells is through the fluorescence [4,3]. To use MACS in cell isolation, prior knowledge about the target cells are also needed.…”

“…Among the currently available techniques for cell isolation, FACS (fluorescence activated cell sorting), MACS (magnetic activated cell sorting), laser micro dissection, manual cell picking are the methods that are the most widely used. [3]. FACS is the most commonly used method but, to use FACS to isolate target cells, complete knowledge about the surface proteins of the target cells and other non-target cells in the cell mixture is needed.…”