3-D polymeric microfluidic devices for BioMOEMS applications

11 páginas, 12 figuras.-- Trabajo presentado en la conferencia "Bioengineered and Bioinspired Systems II"; Sevilla (España); 09-Mayo-2005; Editores: Ricardo A. Carmona, Gustavo Linan-Cembrano.This paper describes the fabrication, packaging and characterization of novel multilayer polymer microfluidic systems fabricated by a CMOS compatible process. These microfluidic devices were specially designed for BioMOEMS applications. Embedded multilayer rectangular smooth and uniform microchannels, 50 to 150 mm wide and 18mm deep were studied. Steady-state flow rates and pressure driven flow control were measured in the laminar flow regime. Flow rates ranging from 1 to 100 µl/min, at pressure drop ranging from 10 to 600 kPa, were obtained. These flow rates yield Reynolds numbers (Re) up to 20. Results indicate that the experimental Re and the flow friction coefficient (f) are in good agreement with the laminar flow theory. These experimental results facilitate the future designs of different microfluidic devices designed by using classical fluidic theory. We also present two different methods developed for macro/microfluidic packaging in order to connect these microfluidic devices to the macroscopic world. The microsystem packaging can withstand pressure drops up from 500 to 2000 kPa with any liquid leakage.This research is sponsored by the Basque and Spanish Governments, under the Torres Quevedo Spanish Fellowship for industrial research and the strategic research program on micro and nanotechnologies (MICROGUNE).Peer reviewe

Optimizing polymer lab-on-chip platforms for ultrasonic manipulation: Influence of the substrate

The choice of substrate material in a chip that combines ultrasound with microfluidics for handling biological and synthetic microparticles can have a profound effect on the performance of the device. This is due to the high surface-to-volume ratio that exists within such small structures and acquires particular relevance in polymer-based resonators with 3D standing waves. This paper presents three chips developed to perform particle flow-through separation by ultrasound based on a polymeric SU-8 layer containing channelization over three different substrates: Polymethyl methacrylate (PMMA); Pyrex; and a cracked PMMA composite-like structure. Through direct observations of polystyrene microbeads inside the channel, the three checked chips exhibit their potential as disposable continuous concentration devices with different spatial pressure patterns at frequencies of resonance close to 1 Mhz. Chips with Pyrex and cracked PMMA substrates show restrictions on the number of pressure nodes established in the channel associated with the inhibition of 3D modes in the solid structure. The glass-substrate chip presents some advantages associated with lower energy requirements to collect particles. According to the results, the use of polymer-based chips with rigid substrates can be advantageous for applications that require short treatment times (clinical tests handling human samples) and low-cost fabrication. © 2015 by the authors; licensee MDPI, Basel, Switzerland.The study has been performed in the framework of two Spanish National Research Project BIO2011-30535-C04-01,02,03, “Development of a high throughput for isolation of tumor cells circulating in peripheral blood”.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer Reviewe

Development of a three-dimensional cell culture system based on microfluidics for nuclear magnetic resonance and optical monitoring.

A new microfluidic cell culture device compatible with real-time nuclear magnetic resonance (NMR) is presented here. The intended application is the long-term monitoring of 3D cell cultures by several techniques. The system has been designed to fit inside commercially available NMR equipment to obtain maximum readout resolution when working with small samples. Moreover, the microfluidic device integrates a fibre-optic-based sensor to monitor parameters such as oxygen, pH, or temperature during NMR monitoring, and it also allows the use of optical microscopy techniques such as confocal fluorescence microscopy. This manuscript reports the initial trials culturing neurospheres inside the microchamber of this device and the preliminary images and spatially localised spectra obtained by NMR. The images show the presence of a necrotic area in the interior of the neurospheres, as is frequently observed in histological preparations; this phenomenon appears whenever the distance between the cells and fresh nutrients impairs the diffusion of oxygen. Moreover, the spectra acquired in a volume of 8 nl inside the neurosphereshow an accumulation of lactate and lipids, which are indicative of anoxic condi-tions. Additionally, a basis for general temperature control and monitoring and a graphical control software have been developed and are also described. The complete platform will allow biomedical assays of therapeutic agents to be performed in the early phases of therapeutic development. Thus, small quantities of drugs or advanced nanodevices may be studied long-term under simulated living conditions that mimic the flow and distribution of nutrient

SU-8 Based Microdevices to Study Self-Induced Chemotaxis in 3D Microenvironments

Tissues are complex three-dimensional structures in which cell behavior is frequently guided by chemotactic signals. Although starvation and nutrient restriction induce many different chemotactic processes, the recreation of such conditions in vitro remains difficult when using standard cell culture equipment. Recently, microfluidic techniques have arisen as powerful tools to mimic such physiological conditions. In this context, microfluidic three-dimensional cell culture systems require precise control of cell/hydrogel location because samples need to be placed within a microchamber without obstruction of surrounding elements. In this article, SU-8 is studied as structural material for the fabrication of complex cell culture microdevices due to its good mechanical properties and sensor integration capacity. Moreover, SU-8 physical properties and their effect on a successful design for precise control of hydrogel location within microfluidic devices are studied. In particular, this manuscript presents a SU-8 based microdevice designed to create “self-induced” medium starvation, based on the combination of nutrient restriction and natural cell metabolism. Results show a natural migratory response toward nutrient source, showing how cells adapt to their own microenvironment modifications. The presented results demonstrate the SU-8 potential for microdevice fabrication applied to cell culture

Glioblastoma on a microfluidic chip: Generating pseudopalisades and enhancing aggressiveness through blood vessel obstruction events

Background: Glioblastoma (GBM) is one of the most lethal tumor types. Hypercellular regions, named pseudo- palisades, are characteristic in these tumors and have been hypothesized to be waves of migrating glioblastoma cells.These “waves” of cells are thought to be induced by oxygen and nutrient depletion caused by tumor-induced blood vessel occlusion. Although the universal presence of these structures in GBM tumors suggests that they may play an instrumental role in GBM’s spread and invasion, the recreation of these structures in vitro has remained challenging. Methods: Here we present a new microfluidic model of GBM that mimics the dynamics of pseudopalisade forma- tion.To do this, we embedded U-251 MG cells within a collagen hydrogel in a custom-designed microfluidic device. By controlling the medium flow through lateral microchannels, we can mimic and control blood-vessel obstruction events associated with this disease. Results: Through the use of this new system, we show that nutrient and oxygen starvation triggers a strong migratory process leading to pseudopalisade generation in vitro.These results validate the hypothesis of pseudo- palisade formation and show an excellent agreement with a systems-biology model based on a hypoxia-driven phenomenon. Conclusions: This paper shows the potential of microfluidic devices as advanced artificial systems capable of mod- eling in vivo nutrient and oxygen gradients during tumor evolution