Single Cell Isolation and Analysis

Abstract

Individual cell heterogeneity within a population can be critical to its peculiar function and fate. Subpopulations studies with mixed mutants and wild types may not be as informative regarding which cell responds to which drugs or clinical treatments. Cell to cell differences in RNA transcripts and protein expression can be key to answering questions in cancer, neurobiology, stem cell biology, immunology, and developmental biology. Conventional cell-based assays mainly analyze the average responses from a population of cells, without regarding individual cell phenotypes. To better understand the variations from cell to cell, scientists need to use single cell analyses to provide more detailed information for therapeutic decision making in precision medicine. In this review, we focus on the recent developments in single cell isolation and analysis, which include technologies, analyses and main applications. Here, we summarize the historical background, limitations, applications, and potential of single cell isolation technologies.

Achieving greater efficiency and higher confidence in single‐cell cloning by combining cell printing and plate imaging technologies

“…Fluorescence‐activated cell sorting (FACS) is another widely used method for single‐cell cloning of clinical cell lines . FACS can efficiently evaluate many cells at the single‐cell level and allows for high‐throughput isolation of single‐cells from subpopulations based on size, granularity, and fluorescence .…”

“…Fluorescence-activated cell sorting (FACS) is another widely used method for single-cell cloning of clinical cell lines. 11,12 FACS can efficiently evaluate many cells at the single-cell level and allows for high-throughput isolation of single-cells from subpopulations based on size, granularity, and fluorescence. 2,13 Cells of interest are passed through a hydrodynamically focused stream of sheath fluid, selected by their light scattering or emission properties, charged by electrical plates and deflected into a collection container such as a multi-well plate.…”

Luminescent nanomaterials for droplet tracking in a microfluidic trapping array

“…[6, 7] However, flow cytometry is often limited by its need for large sample size, the cost and size of the instrument,[8] and the use of rapid flow in the system which, when coupled with non-specific surface markers, can negatively affect cell viability. [9] There have been reports on spectral crosstalk between these fluorescent-cell barcodes which was addressed by mass cytometry, a method where each sample is labelled with a unique combination of lanthanide isotopes (called mass-tag barcoding). [6, 10] Despite having a lower throughput than flow cytometry,[4] the spectrally distinct mass-tag barcodes in mass cytometry make it an ideal system for multiplex studies.…”

Study on Cell-Capturing Microfluidic Device in High Flow Rates through Controlling Shape of Microstructures and Their Alignments

“…Even though some capturing schemes have been reported, passive methods using hydrodynamic weirs seem to be attractive because their implementation is simple and does not need expertise for sophisticated and complex microfluidic control [4]. However, most weir or post systems have relatively poor capturing efficiency, it is needed to enhance the designs and fabrication methods in order to apply for the rare cells populations study [5][6][7]. Considering the rare cells isolation rate using microfluidic devices is approaching to about 90%, the sequential capturing single cells is also becoming important.…”

“…Considering the rare cells isolation rate using microfluidic devices is approaching to about 90%, the sequential capturing single cells is also becoming important. Thus, we have presented the a rare cellscapturing device with three different geometries to investigate the effects of geometries on capturing rates in a very high flow rate of 300 µ L/min, to meet with the condition of over 7.5 mL/h (125 µ L /min) of a FDA-approved machine [6].…”