Reproducibility and Robustness of a Real-Time Microfluidic Cell Toxicity Assay
- 20 April 2011
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 83 (10), 3890-3896
- https://doi.org/10.1021/ac200273f
Abstract
Numerous opportunities exist to apply microfluidic technology to high-throughput and high-content cell-based assays. However, maximizing the value of microfluidic assays for applications such as drug discovery, screening, or toxicity evaluation will require assurance of within-device repeatability, day-to-day reproducibility, and robustness to variations in conditions that might occur from laboratory to laboratory. This report describes a study of the performance and variability of a cell-based toxicity assay in microfluidic devices made of poly(dimethylsiloxane) (PDMS). The assay involves expression of destabilized green fluorescent protein (GFP) as a reporter of intracellular protein synthesis and degradation. Reduction in cellular GFP due to inhibition of ribosome activity by cycloheximide (CHX) was quantified with real-time quantitative fluorescence imaging. Assay repeatability was measured within a 64-chamber microfluidic device. Assay performance across a range of cell loading densities within a single device was assessed, as was replication of measurements in microfluidic devices prepared on different days. Assay robustness was tested using different fluorescence illumination sources and reservoir-to-device tubing choices. Both microfluidic and larger scale assay conditions showed comparable GFP decay rates upon CHX exposure, but the microfluidic data provided the higher level of confidence.Keywords
This publication has 28 references indexed in Scilit:
- Fundamentals of microfluidic cell culture in controlled microenvironmentsChemical Society Reviews, 2010
- Biological implications of polydimethylsiloxane-based microfluidic cell cultureLab on a Chip, 2009
- Microfluidics meet cell biology: bridging the gap by validation and application of microscale techniques for cell biological assaysBioEssays, 2008
- Live cells‐based cytotoxic sensorchip fabricated in a microfluidic systemBiotechnology & Bioengineering, 2007
- Quantitative Comparison between Microfluidic and Microtiter Plate Formats for Cell-Based AssaysAnalytical Chemistry, 2007
- Automated live cell imaging of green fluorescent protein degradation in individual fibroblastsCytometry Part A, 2007
- Understanding microchannel culture: parameters involved in soluble factor signalingLab on a Chip, 2007
- Diffusion dependent cell behavior in microenvironmentsLab on a Chip, 2005
- Microenvironment design considerations for cellular scale studiesLab on a Chip, 2004
- Generation of Destabilized Green Fluorescent Protein as a Transcription ReporterJournal of Biological Chemistry, 1998