A Novel Long-term, Multi-Channel and Non-invasive Electrophysiology Platform for Zebrafish.

Zebrafish are a popular vertebrate model for human neurological disorders and drug discovery. Although fecundity, breeding convenience, genetic homology and optical transparency have been key advantages, laborious and invasive procedures are required for electrophysiological studies. Using an electrode-integrated microfluidic system, here we demonstrate a novel multichannel electrophysiology unit to record multiple zebrafish. This platform allows spontaneous alignment of zebrafish and maintains, over days, close contact between head and multiple surface electrodes, enabling non-invasive long-term electroencephalographic recording. First, we demonstrate that electrographic seizure events, induced by pentylenetetrazole, can be reliably distinguished from eye or tail movement artifacts, and quantifiably identified with our unique algorithm. Second, we show long-term monitoring during epileptogenic progression in a scn1lab mutant recapitulating human Dravet syndrome. Third, we provide an example of cross-over pharmacology antiepileptic drug testing. Such promising features of this integrated microfluidic platform will greatly facilitate high-throughput drug screening and electrophysiological characterization of epileptic zebrafish

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)

Liquid recirculation in microfluidic channels by the interplay of capillary and centrifugal forces

We demonstrate a technique to recirculate liquids in a microfluidic device, maintaining a thin fluid layer such that typical diffusion times for analytes to reach the device surface are < 1 min. Fluids can be recirculated at least 1000 times across the same surface region, with no change other than slight evaporation, by alternating the predominance of centrifugal and capillary forces. Mounted on a rotational platform, the device consists of two hydrophilic layers separated by a thin pressure-sensitive adhesive (PSA) layer that defines the microfluidic structure. We demonstrate rapid, effective fluid mixing with this device

Optically addressable single-use microfluidic valves by laser printer lithography

We report the design, fabrication, and characterization of practical optofluidic valves fabricated using laser printer lithography. Valves are opened by directing optical energy from a solid-state laser, with similar power characterisitcs to those used in CD/DVD drives, to a spot of printed toner where localized heating melts an orifice in the polymer layer in as little as 500 ms, connecting previously isolated fluidic components or compartments. Valve functionality, response time, and laser input energy dependence of orifice size are reported for cyclo-olefin copolymer (COC) and polyethylene terephthalate (PET) films. Implementation of these optofluidic valves is demonstrated on pressure-driven and centrifugal microfluidic platforms. In addition, these “one-shot” valves comprise a continuous polymer film that hermetically isolates on-chip fluid volumes within fluidic devices using low-vapor-permeability materials; we confirmed this for a period of one month. The fabrication and integration of optofluidic valves is compatible with a range of polymer microfabrication technologies and should facilitate the development of fully integrated, reconfigurable, and automated lab-on-a-chip systems, particularly when reagents must be stored on chip for extended periods, e.g. for medical diagnostic devices, lab-on-a-chip synthetic systems, or hazardous biochemical analysis platforms

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

Integrating Nanomembrane Separation with Plasmonic Detection for Real-Time Cell Culture Monitoring

To further understand cellular responses to drug treatment the dynamics of a reduced secretome shall be investigated. Currently there is no method for the detection of secreted small molecules in real time, label-free and with a high resolution. We present a novel design, which integrates nanopore filtration technology with highly sensitive plasmonic detection that allows real time monitoring of filtered molecules with a high spatial resolution and label free. The cell culture chamber is separated from the site of detection only by our biocompatible nanomembrane filter with a thickness of less than 100 nm to exclude the majority of background signals from the cell culture. The fast filtration of the cell culture constituents through the nanomembrane to the detector allows the observation of the dynamics of secreted molecules during cell culture and/or drug application. The setup offers new possibilities for drug screening and cell assays and may reveal new insights into cell signaling and drug responses. This setup shall be used to monitor cell culture or tissue culture without the necessity of labeling. This can be particularly important for the very popular “organ-on-a-chip” or “patient-on-a-chip” approaches to monitor tissue reactions to drug treatments with a high spatial resolution. Please click Additional Files below to see the full abstract