Configurable and Up-Scalable Microfluidic Life Science Platform for Cell Based Assays by Gravity Driven Sequential Perfusion and Diffusion

Microfluidics has the potential to significantly change the way modern biology is performed, but for this potential to be realized several on-chip integration and operation challenges have to be addressed. Critical issues are addressed in this work by first demonstrating an integrated microfluidic tmRNA purification and real time nucleic acid sequence based amplification (NASBA) device. The device is manufactured using soft lithography and a unique silica bead immobilization method for the nucleic acid micro purification column. The integrated device produced a pathogen-specific response in < 3 min from the chip-purified RNA. Further enhancements in the device design and operation that allow the on-chip integration of mammalian cell handling and culturing produced a novel integrated NASBA array. This system demonstrated for the first time that it is possible to combine on a single micro-device cell culture and real time NASBA. In order to expand the cell based assay capabilities of the integrated NASBA array and simplify the device operation novel hydrodynamics and cell sedimentation within trench structures and gravity driven sequential perfusion and diffusion mechanisms were developed. These mechanisms were characterized and implemented within an iCell array device. iCell array can completely integrate cell based assays with bio-analytical read-out. The device is highly scalable and can enable the configurable on-chip integration of procedures such as adherent and non-adherent cell-culture, cellstimulation, cell-lysis, cell-fixing, protein-immunoassays, bright field and fluorescent microscopic monitoring, and real time detection of nucleic acid amplification. The device uses on-board gravity driven flow control which makes it simple and economical to operate with dilute samples (down to 5 cells per reaction), low reagent volumes (50 nL per reaction), highly efficient cell capture (100% capture rates) and single cell protein and gene expression sensitivity. The key results from this work demonstrate a novel technology for versatile, fully integrated microfluidic array platforms. By multiplexing this integrated functionality, the device can be used from routine applications in a biology laboratory to high content screenings

Hybrid integrated platform of PDMS microfluidics and silica capillary for effective CE and ESI-MS coupling

We present an effective hybrid integration of PDMS microfluidic devices and fused silica capillaries. These hybrid microfluidic integrated PDMS and silica capillary (iPSC) modules exhibit a novel architecture and method for leakage free CE sample injection requiring only a single high voltage source. Use of the iPSC devices is based on a modular approach which allows the capillary to be reused over 1,000 times whilst replacing the fluidics below it for different experiments. Integrating fused silica capillaries with PDMS microfluidics allows the direct application of a wide variety of well established conventional CE protocols for complex analyte separations and ESI-MS coupling, allowing users to focus on the sample analysis rather than the development of new separation protocols. The iPSC fabrication method is simple (3 steps) and quick (7 min)

Monolithic centrifugal microfluidic platform for bacteria capture and concentration, lysis, nucleic-acid amplification, and real-time detection

We report the design, fabrication, and characterization of a polymer centrifugal microfluidic system for the specific detection of bacterial pathogens. This single-cartridge platform integrates bacteria capture and concentration, supernatant solution removal, lysis, and nucleic-acid sequence-based amplification (NASBA) in a single unit. The unit is fabricated using multilayer lamination and consists of five different polymer layers. Bacteria capture and concentration are accomplished by sedimentation in five minutes. Centrifugation forces also drive the subsequent steps. A wax valve is integrated in the cartridge to enable high-speed centrifugation. Oil is used to prevent evaporation during reactions requiring thermal cycling. Device functionality was demonstrated by real-time detection of E. coli from a 200-muL sample

Thin film diffusion barrier formation in PDMS microcavities

We describe a method to form glass like thin film barrier in polydimethylsiloxane (PDMS) microcavities. The reactive fragments for the surface reaction were created from O2 and hexamethyldisiloxane (HMDS) in RF plasma environment. The reaction is based on migration of the reactive fragments into the microcavities by diffusion, to form a glass like thin film barrier to conceal the naked surface of PDMS. The barrier successfully blocked penetration of a fluorescent dye rhodamine B (RhB) into PDMS. The thickness of the barrier could be controlled by the time of reaction and the pressure inside the reaction chamber. There is a wide range of applications of such a technique in various fields, e.g. for coating the covered surfaces of microfluidic channels, tubes, capillaries, medical devices, catheters, as well as chip-integrated capillary electrophoresis and advanced electronic and opto-fluidic packaging

Lab-in-a-trench platform for real-time monitoring of cell surface protein expression

In this work we for the first time demonstrate real-time monitoring of the expression of membrane proteins in native, live cells, free of hydrodynamic stress at single cell resolution. This micro-optofluidic mechanism is uniquely enabled by the intricate interplay of gravity induced sedimentation with laminar flow, fast diffusion and short optical path length on our lab-in-a-trench platform