title: A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay.
abstract: A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single infected host cells into the individual micro-wells of the biochip was achieved by one-step vacuum-driven microfluidics. The adhered bacterial cells were then quantified by direct on-chip real-time PCR assay with single-bacterium-detection sensitivity. The performance of this microfluidic platform was demonstrated through profiling of the association of a common bacterial pathogen, Pseudomonas aeruginosa, to single host human lung epithelial A549 cells, revealing an adherence distribution that has not been previously reported. This microfluidic platform offers a simple and effective tool for biologists to analyze pathogen-host interaction at the single-cell level without the necessities of fluorescence labeling. The chip can similarly be used for other PCR-based applications requiring single-cell analysis.
authors: School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore, Singapore.
authors: Rui
authors: R
authors: Zhang
authors: Hai-Qing
authors: HQ
authors: Gong
authors: Xu Dong
authors: XD
authors: Zeng
authors: Chun Chau
authors: CC
authors: Sze
mesh_terms: D001422:Bacterial Adhesion; D002460:Cell Line; D054884:Host-Pathogen Interactions; D006801:Humans; D008168:Lung / Q000166:cytology*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation*; D059010:Single-Cell Analysis / Q000295:instrumentation*