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Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells
Published on: February 2, 2013
Microfluidic single-cell array cytometry for the analysis of tumor apoptosis
Donald Wlodkowic1, Shannon Faley, Michele Zagnoni
1The Bioelectronics Research Centre, Department of Electronics and Electrical Engineering, University of Glasgow, G12 8LT, United Kingdom.
Abstract:
Limitations imposed by conventional analytical technologies for cell biology, such as flow cytometry or microplate imaging, are often prohibitive for the kinetic analysis of single-cell responses to therapeutic compounds. In this paper, we describe the application of a microfluidic array to the real-time screening of anticancer drugs against arrays of single cells. The microfluidic platform comprises an array of micromechanical traps, designed to passively corral individual nonadherent cells. This platform, fabricated in the biologically compatible elastomer poly(dimethylsiloxane), PDMS, enables hydrodynamic trapping of cells in low shear stress zones, enabling time-lapse studies of nonadherent hematopoietic cells. Results indicate that these live-cell, microfluidic microarrays can be readily applied to kinetic analysis of investigational anticancer agents in hematopoietic cancer cells, providing new opportunities for automated microarray cytometry and higher-throughput screening. We also demonstrate the ability to quantify on-chip the anticancer drug induced apoptosis. Specifically, we show that with small numbers of trapped cells (approximately 300) under careful serial observation we can achieve results with only slightly greater statistical spread than can be obtained with single-pass flow cytometer measurements of 15,000-30,000 cells.
Insights
This study introduces a microfluidic array for real-time anticancer drug screening on single hematopoietic cells. The platform enables kinetic analysis and apoptosis quantification, offering a high-throughput alternative to conventional methods.
Area of Science:
- Cell biology
- Microfluidics
- Pharmacology
Background:
- Conventional cell analysis methods like flow cytometry have limitations for single-cell kinetic studies.
- Analyzing single-cell responses to therapeutic compounds requires advanced technologies.
Purpose of the Study:
- To apply a microfluidic array for real-time screening of anticancer drugs against single cells.
- To enable kinetic analysis and apoptosis quantification of drug effects on hematopoietic cells.
Main Methods:
- Fabrication of a microfluidic platform using poly(dimethylsiloxane) (PDMS).
- Utilizing micromechanical traps for hydrodynamic trapping of nonadherent cells.
- Performing time-lapse studies for kinetic analysis of drug responses.
Main Results:
- Demonstrated real-time screening of anticancer agents against single hematopoietic cells.
- Successfully quantified anticancer drug-induced apoptosis on-chip.
- Achieved comparable statistical results with ~300 cells compared to 15,000-30,000 cells in flow cytometry.
Conclusions:
- Microfluidic microarrays provide a viable platform for kinetic analysis of investigational anticancer agents.
- This technology offers opportunities for automated microarray cytometry and higher-throughput screening.
- Enables precise quantification of drug-induced apoptosis with minimal cell numbers.
