An Inexpensive Microfluidic PDMS Chip for Visual Detection of Biofilm-forming Bacteria

摘要

Aims: Design and assembly of an inexpensive microfluidic PDMS chip for visual detection of cell adhesion and biofilm formation. Study Design: Three different styles of microchannels (2.6, 5.0, and 11.5 μl volumes) were designed, fabricated and tested for adhesion and biofilm formation in a microfluidic system. The pressure drop measurements system includes a bio-Ferrograph connected to the PDMS microchannel via a syringe and a pressure transducer. Methodology: Microfluidic chips were fabricated using Polydimethylsiloxane (PDMS) by means of soft lithography. Different cell densities of E.coli K12 cells were introduced to investigate adhesion and biofilm formation at different time intervals. Stabilization time and hydraulic resistance were obtained via a Bio-Ferrograph connected to a pressure transducer. Results: PDMS microfluidic volume (2.6 μl) failed to generate noticeable biofilm, while slight and greatest yield occurred with PDMS microchannels (5.0, and 11.5 μl), respectively, and could detect as low as 26 cells in 11.5 μl microchannel. As incubation time and/or initial cell density increases, cell adhesion increased, illustrated by crystal violet color intensity. High stabilization time (3 h) didn’t allow for bacterial attachment and cultivation inside the microchannel (2.6 μl) while lower stabilization time (10 min) yielded the highest capacity of cell adhesion in microchannel (11.5 μl). Conclusions: We developed a microfluidic chip with low stabilization time and hydraulic resistance, thus offering more volume for adhesion of bacterial cells and biofilm formation. It allowed bacterial cultivation without any addition of nutrients. The microfluidic chip provides a platform to monitor biofilm growth and can be integrated in situ investigations for biological systems, food biotechnology and other industrial biotechnology applications. This would allow a non-destructive and non-invasive monitoring of the biofilm-forming bacteria inside the PDMS microfluidic chip. This work opens opportunities for further investigations of pressure drop phenomena in microchannels that would otherwise go unnoticed in macro scale measurements.