title: Development of a microfluidic biochip for online monitoring of fungal biofilm dynamics.
abstract: Microfabricated biochips are developed to continuously monitor cell population dynamics in a non-invasive manner. In the presented work we describe the novel combination of contact-less dielectric microsensors and microfluidics to promote biofilm formation for quantitative cell analysis. The cell chip consists of a polymeric fluidic (PDMS) system bonded to a glass wafer containing the electrodes while temperature and fluid flow are controlled by external heating and pumping stations. The high-density interdigitated capacitors (microIDES) are isolated by a 550 nm multi-passivation layer of defined dielectric property and provide stable, robust and non-drifting measurement conditions. The performance of this detector is evaluated using various bacterial and yeast strains. The high sensitivity of the developed dielectric microsensors allows direct identification of microbial strains based on morphological differences and biological composition. The novel biofilm analysis platform is used to continuously monitor the dynamic responses of C. albicans and P. pastoris biofilms to increased shear stress and antimicrobial agent concentration. While the presence of shear stress triggers significant changes in yeast growth profiles, the addition of 0.5 microg mL(-1) amphotericin B revealed two distinct dynamic behaviors of the C. albicans biofilm. Initially, impedance spectra increased linearly at 30 Omega h(-1) for two hours followed by 10 Omega h(-1) (at 50 kHz) over 10 hours while cell viability remained above 95% during fungicide administration. These results demonstrate the ability to directly monitor dielectric changes of sub-cellular components within a living cell population.
authors.affiliation: Division of Nano-System-Technologies, Austrian Research Centers GmbH-ARC, Donau-City-Street 1, 1220 Vienna, Austria.
authors.forename: Lukas
authors.initials: L
authors.lastname: Richter
authors.forename: Christoph
authors.initials: C
authors.lastname: Stepper
authors.forename: Andy
authors.initials: A
authors.lastname: Mak
authors.forename: Alessa
authors.initials: A
authors.lastname: Reinthaler
authors.forename: Rudolf
authors.initials: R
authors.lastname: Heer
authors.forename: Michael
authors.initials: M
authors.lastname: Kast
authors.forename: Hubert
authors.initials: H
authors.lastname: Brückl
authors.forename: Peter
authors.initials: P
authors.lastname: Ertl
mesh_terms: D018441:Biofilms; D002462:Cell Membrane; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D009862:Online Systems / Q000295:instrumentation*