Microfluidic Single-Cell Array Cytometry for the Analysis of Tumor Apoptosis

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 anti-cancer drugs against arrays of single cells. The microfluidic platform comprises an array of micromechanical traps, designed to passively corral individual non-adherent 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 non-adherent hematopoietic cells. Results indicate that these live-cell, microfluidic microarrays can be readily applied to kinetic analysis of investigational anti-cancer 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 anti-cancer drug induced apoptosis. Specifically, we show that with small numbers of trapped cells (~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.

Single-cell attachment and culture method using a photochemical reaction in a closed microfluidic system

“…13 Numerous microfluidic-based single-cell analysis methods have been developed and can be classified according to the positioning methods, such as electrical positioning, 14 hydrodynamic trapping, 15 physical trapping, 16,17 and microwell trapping. 18,19 However, these methods are inappropriate for the detection of environmental factors such as cell-cell contact and cell-extracellular matrix ͑ECM͒ contact. Cell contact on the surface plays a pivotal role in the behavior of adherent cells, as adhesion triggers intracellular signal transduction, affecting cellular growth, differentiation, and proliferation.…”

Streamline based design guideline for deterministic microfluidic hydrodynamic single cell traps

“…This streamline design guideline may also be applied to trapping arrays. 12,23,24 By optimizing the design such that all relevant streamlines are captured, multiple targets of interest may be captured in a high-throughput manner without any loss provided that the number of traps exceeds the number of expected targets.…”

Microfluidics-Based Cell Manipulation and Analysis

“…Meanwhile, microfluidic devices can realise biological experiments in a high-throughput way, while being based on the miniaturising macroscopic systems and taking advantage of massive parallel processing. Thus far, microfluidic applications have been involved in many experimental parts of cell manipulation and analysis, such as cell trapping/sorting, cell culture/co-culture, cytotoxicity, PCR, DNA sequencing, and gene analyses (VelveCasquillas et al, 2010;Wlodkowic et al, 2009;Melin et al, 2007). Furthermore, a large number of novel microfluidic devices have been reported for cell research and tissue simulation in last 10 years (Ho et al, 2006;Huh et al, 2010;Sung et al, 2011).…”