Mahtab Hassanpourfard

Protocol for Biofilm Streamer Formation in a Microfluidic Device with Micro-pillars

Several bacterial species possess the ability to attach to surfaces and colonize them in the form of thin films called biofilms. Biofilms that grow in porous media are relevant to several industrial and environmental processes such as wastewater treatment and CO2 sequestration. We used Pseudomonas fluorescens, a Gram-negative aerobic bacterium, to investigate biofilm formation in a microfluidic device that mimics porous media. The microfluidic device consists of an array of micro-posts, which were fabricated using soft-lithography. Subsequently, biofilm formation in these devices with flow was investigated and we demonstrate the formation of filamentous biofilms known as streamers in our device. The detailed protocols for fabrication and assembly of microfluidic device are provided here along with the bacterial culture protocols. Detailed procedures for experimentation with the microfluidic device are also presented along with representative results.

Rapid label-free detection of E. coli using antimicrobial peptide assisted impedance spectroscopy

There is an increasing demand for rapid detection of waterborne pathogens to monitor drinking water safety. We demonstrate a compact, label-free sensor array for rapid detection of Escherichia coli (E. coli) in contaminated water samples using antimicrobial peptide assisted impedimetric sensor platform. Interdigitated electrode arrays immobilized with the antimicrobial peptide Colicin V (ColV) were used to screen the affinity towards different bacterial strains by monitoring impedance variations in real-time. This ColV assisted impedance biosensor exhibited high selectivity towards Gram-negative strains particularly towards E. coli strains. This selective detection of E. coli from other strains was observed at 102 cfu mL−1, which is clinically relevant. The sensor can detect E. coli from 102 to 106 cfu mL−1 in water sample at pH 7 to 9. These results show that the antimicrobial peptide ColV assisted impedimetric array is capable of rapid, specific detection of E. coli in contaminated water samples.

Portable Nanofiber-Light Addressable Potentiometric Sensor for Rapid Escherichia coli Detection in Orange Juice

The growing need to prevent pathogen outbreaks is irrefutable in the case of the food industry. Early detection in products, especially beverages, contaminated with bacterial strains is vital to avoid infected foods from reaching the consumer. If E. coli is pesent in such foods, it can cause infections. It can also be an indicator of the existence of other harmful coliforms. In this study, we have investigated the detection of Escherichia coli ( E. coli) in orange juice using a portable nanofiber-light addressable potentiometric sensor (NF-LAPS). We have chosen electrospun pH-sensitive poly(vinyl alcohol)/poly(acrylic acid) (PVA/PAA) hydrogel NFs as the sensitive layer. The successful detection of E. coli was reported with the NF-LAPS in less than 1 h. The limit of detection (LOD) measured in the sensor is found to be102 CFU/mL. We have confirmed the selectivity of the biosensor toward E. coli by examining the response of the NF-LAPS against Salmonella typhimurium ( S. typhi), also commonly found in orange juice. Despite the complex nature of orange juice, the response of the biosensor is in no way affected while orange juice is tested as is.

Biofilm Streamer Formation in a Microfluidic Porous Media Mimic

Several bacterial species possess the ability to attach to surfaces and colonize themselves in thin films called biofilms. Biofilms that grow in porous media are relevant to several industrial and environmental processes such as wastewater treatment and CO2 sequestration. We used Pseudomonas fluorescens, a gram negative aerobic biofilm forming bacteria, to investigate biofilm formation in a microfluidic porous media mimic device. The microfluidic device consists of an array of micro-posts, which were fabricated using soft-lithography. Subsequently, biofilm formation in this device was investigated as a function of time and the formation of filamentous biofilms known as streamers was observed. Furthermore, we used computational fluid mechanics simulation to better understanding of the streamer formation.© 2014 ASME