Parallel single‐cell analysis microfluidic platform

Abstract

We report a PDMS microfluidic platform for parallel single-cell analysis (PaSCAl) as a powerful tool to decipher the heterogeneity found in cell populations. Cells are trapped individually in dedicated pockets, and thereafter, a number of invasive or non-invasive analysis schemes are performed. First, we report single-cell trapping in a fast (2-5 min) and reproducible manner with a single-cell capture yield of 85% using two cell lines (P3x63Ag8 and MCF-7), employing a protocol which is scalable and easily amenable to automation. Following this, a mixed population of P3x63Ag8 and MCF-7 cells is stained in situ using the nucleic acid probe (Hoechst) and a phycoerythrin-labeled monoclonal antibody directed at EpCAM present on the surface of the breast cancer cells MCF-7 and absent on the myeloma cells P3x63Ag8 to illustrate the potential of the device to analyze cell population heterogeneity. Next, cells are porated in situ using chemicals in a reversible (digitonin) or irreversible way (lithium dodecyl sulfate). This is visualized by the transportation of fluorescent dyes through the membrane (propidium iodide and calcein). Finally, an electrical protocol is developed for combined cell permeabilization and electroosmotic flow (EOF)-based extraction of the cell content. It is validated here using calcein-loaded cells and visualized through the progressive recovery of calcein in the side channels, indicating successful retrieval of individual cell content.

Review on Impedance Detection of Cellular Responses in Micro/Nano Environment

“…Due to their miniaturization and automation, there are a number of advantages of using microfluidic systems, such as less sample/reagent consumption, reduced risk of contamination, less cost per analysis, lower power consumption, enhanced sensitivity and specificity, and higher reliability. Microfluidic systems have been developed for various biological analytical applications, such as DNA analysis [4][5][6][7][8], immunoassay [9][10][11][12][13], and cell analysis [14][15][16][17][18]. Moreover, a number of demonstrations showed that cell culture can be performed on the microfluidic systems to achieve higher throughput and more reliable results [19,20].…”

Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

“…Several other methods have also been coupled with microfluidics for cell immobilization and conducting controlled, complete cell assays. Flow-based active cell trapping by using control valves [14,15], non-invasive optical trapping [16][17][18], dielectrophoresis [19][20][21], surface chemistry modification techniques [22,23], arrays of physical barriers [24], cell trapping by negative pressure [25,26], and hydrodynamic methods [27][28][29] are some of these successfully established techniques.…”

Droplet Microfluidics for Screening of Surface-Marker and Secretory Protein Expression

“…This system has shown great potential not only in sample manipulation and handling but also created a reputation for itself in being able to integrate multiparametric analysis in single platforms. LOC microfluidic devices have found a major niche in various facets of biochemical research including drug discovery [2,3], diagnostics [4][5][6][7][8], single cell analysis [9][10][11], single molecule detection [12][13][14], chemical synthesis [15,16], and other biological and chemical assays [17][18][19][20].…”