Polymer based microfabrication and its applications in optical MEMS and bioMEMS

Due to its ease of fabrication, low cost and great variety of functionalities, polymer has become an important material in microfabrication. MEMS devices with polymer as the structure material have found applications in various fields, especially in BioMEMS and optical MEMS. In this dissertation, three polymer based microfabricated devices/components have been developed and tested. Various polymer based fabrication techniques, such as high aspect ratio SU-8 photolithography, three dimensional polydimethylsiloxane (PDMS) soft lithography, multi-layer soft lithography and PDMS double casting technique have been developed/studied and employed in the device fabrication process. The main contribution of this dissertation includes: (1) Developed two novel methods for the fabrication of out-of-plane microlens. The PDMS and UV curable polymer (NOA73) replication technique made possible the fast replication of out-of-plane microlens and broaden the lens material selection. The in-situ pneumatical microlens fabrication technique, on the other hand, provides feasible method to integrate out-of-plane microlens into microfluidic chips requiring minimal design footprint and fabrication complexity. (2) Design, fabrication and test of a microchip flow cytometer with 3-D hydrofocusing chamber and integrated out-of-plane microlens as on-chip optical detection component. The developed micro flow cytometer offers 3-D hydrofocusing like conventional cytometer cuvette, and has on-chip microlens for optical signal collection to improve the detection efficiency. With the latest design improvement, the hydrofocusing chamber can focus the sample stream down to less than 10 m in diameter in both vertical and horizontal directions. (3) Development of a PDMS microchip based platform for multiplex immunoassay applications. Integrated micro valves were employed for manipulation of fluidic reagents in the microchannel network. PDMS surface was used as the solid phase substrate for immuno-reactions. Preliminary results showed that, even with low cost polyclonal goat anti-mouse IgG as the reporter antibody, the detection limit of goat mouse IgG can reach as low as 5 ng/mL (about 33 pM). With the continuous advance in microfabrcation technique and polymer science, polymer based microfabrication and polymer MEMS devices will keep to evolve. In the future, more work needs to be done in this field with great potential and endless innovations

Mems (Micro-Electro-Mechanical-Systems) Based Microfluidic Platforms for Magnetic Cell Separation

Microfluidic platforms for magnetic cell separation were developed and investigated for isolation of magnetic particles and magnetically tagged cells from a fluidic sample. Two types of magnetic separation platforms were considered: an Isodynamic Open Gradient Magnetic Sorter (OGMS) and a multistage bio-ferrograph. Miniaturized magnets were designed using magnetostatic simulation software, microfluidic channels were fabricated using microfabrication technology and magnetic separation was investigated using video microscopy and digital image processing. The isodynamic OGMS consisted of an external magnetic circuit and a microfabricated channel (biochip) with embedded magnetic elements. The biochip is placed inside the magnetic field of the external circuit to obtain nearly constant energy density gradient in the portion of the channel used for separation. The microfabrication process involved improving adhesion of thick SU-8 to Pyrex, forming enclosed channels using a low temperature SU-8 adhesive bonding, and fabricating patterned plating molds on both sides of the bonded wafers. Adhesion of SU-8 to Pyrex was improved by using a highly crosslinked thin SU-8 adhesion layer, and enclosed microchannels were fabricated using selectively exposed SU-8 bond formation layers. Electroplating molds were fabricated using KMPR photoresists and were integrated on both sides of the bonded wafers. The multistage bio-ferrograph consisted of a microfabricated enclosed channel placed on the surface of a multi-unit magnet (4 trapezoidal magnets placed in series) assembly such that magnetic cells from a flowing stream would be deposited on designated locations. The OGMS was able to deflect magnetic particles by 500-1000 microns and the capture efficiencies of magnetic particles and cells with the multistage bio-ferrograph were 80-85 percent and 99.5 percent, respectivel

Mems (Micro-Electro-Mechanical-Systems) Based Microfluidic Platforms for Magnetic Cell Separation

Microfluidic platforms for magnetic cell separation were developed and investigated for isolation of magnetic particles and magnetically tagged cells from a fluidic sample. Two types of magnetic separation platforms were considered: an Isodynamic Open Gradient Magnetic Sorter (OGMS) and a multistage bio-ferrograph. Miniaturized magnets were designed using magnetostatic simulation software, microfluidic channels were fabricated using microfabrication technology and magnetic separation was investigated using video microscopy and digital image processing. The isodynamic OGMS consisted of an external magnetic circuit and a microfabricated channel (biochip) with embedded magnetic elements. The biochip is placed inside the magnetic field of the external circuit to obtain nearly constant energy density gradient in the portion of the channel used for separation. The microfabrication process involved improving adhesion of thick SU-8 to Pyrex, forming enclosed channels using a low temperature SU-8 adhesive bonding, and fabricating patterned plating molds on both sides of the bonded wafers. Adhesion of SU-8 to Pyrex was improved by using a highly crosslinked thin SU-8 adhesion layer, and enclosed microchannels were fabricated using selectively exposed SU-8 bond formation layers. Electroplating molds were fabricated using KMPR photoresists and were integrated on both sides of the bonded wafers. The multistage bio-ferrograph consisted of a microfabricated enclosed channel placed on the surface of a multi-unit magnet (4 trapezoidal magnets placed in series) assembly such that magnetic cells from a flowing stream would be deposited on designated locations. The OGMS was able to deflect magnetic particles by 500-1000 microns and the capture efficiencies of magnetic particles and cells with the multistage bio-ferrograph were 80-85 percent and 99.5 percent, respectivel

Development of an integrated microspectrometer using arrayed waveguide grating (AWG)

With non-invasive properties and high sensitivities, portable optical biosensors are extremely desirable for point-of-care (POC) applications. Lab-on-a-chip technology such as microfluidics has been treated as an ideal approach to integrate complex sample processing and analysis units with optical detection elements. Spectroscopic sensing (such as fluorescence, Raman and absorption spectroscopy) remains the most highly developed, widely applied, optical technique. However, conventional spectroscopic sensing systems still rely on bulky and expensive dispersive components such as spectrophotometers in a well established laboratory. The work in this thesis is to develop an integrated dispersive component in combination with a microfluidic chip, providing a portable and inexpensive platform for on-chip spectroscopic sensing. In this study, an arrayed waveguide grating (AWG) design developed for telecommunication is re-engineered and utilized to realise a compact, dispersive optical component operating in the visible spectral region. The AWG devices operating in the visible region (λ_c=680 nm) are designed and fabricated with flame hydrolysis deposited (FHD) silica waveguide material. The micro-spectrometer in this proof of concept study has a small (1 cm x 1 cm) footprint and 8 output channels centred on different wavelengths. A series of fabrication issues and challenges are investigated and discussed for the specific AWG device. Subsequently, a sample cuvette is formed by using lithographic technique and dry etching process. Following this, a PDMS chip with microfluidic channels is bonded with the AWG device, leading to an integrated AWG-microfluidic platform. To the best of the author’s knowledge, this is the first work to integrate a visible AWG device and a microfluidic chip towards spectroscopic sensing. The monolithic integrated AWG microspectrometer–microfluidic platform is demonstrated for fluorescence spectroscopic analysis. Signals from the output channels detected on a camera chip can be used to re-create the complete fluorescence spectrum of an analyte. By making fluorescence measurements of (i) mixed quantum dot solutions, (ii) an organic fluorophore (Cy5) and (iii) the propidium iodide (PI)-DNA assay, the results obtained illustrate the unique advantages of the AWG platform for simultaneous, quantitative multiplex detection and its capability to detect small spectroscopic shifts. Although the current system is designed for fluorescence spectroscopic analysis, in principle, it can be implemented for other types of analysis, such as Raman spectroscopy. Fabricated using established semiconductor industry methods, this miniturised platform holds great potential to create a handheld, low cost biosensor with versatile detection capability. Also, the AWG device design is modified with focusing properties that enable localised spectroscopic measurements. Micro-beads based, multiplexed fluorescence detection is performed with the AWG + CCD system and the results have demonstrated capabilities of using the adapted AWG device for localised, multiplexed fluorescence detections, opening up potential applications in the field of cell sorting and single cell analysis. Furthermore, the AWG-microfluidic device is investigated for absorption spectroscopy measurement. As a test system, the pH dependence of the absorption spectra of bromophenol blue is measured to illustrate how an AWG device could be used as a colorimetric pH sensor. Overall, it is believed that the AWG technology holds great potential to realise a compact, integrated spectroscopic biosensor for point-of-care applications

Design of a microfluidic chip for three dimensional hydrodynamic focusing in cell cytometry applications

The Focusing of cells is an important part of cytometry applications and can be achieved by different techniques. 3D hydrodynamic focusing has generated considerable interest over the last few years, owing to its simplicity and its independence from an electric potential for focusing. Current 3D hydrodynamic focusing devices require multilayer structures necessitating complex fabrication. Moreover, the existing designs show poor efficiencies in focusing. In the present work, three novel 3D hydrodynamic focusing designs consisting of a main channel for sample fluid flow and three pairs of side channels for focusing are proposed and modelled. A novel three dimensional hydrodynamic focusing design is proposed and simulated. In order to develop the numerical model for three dimensional focusing designs, a theoretical review of parameters affecting the fluid flow in microfluidic structure has been performed. Simulation was performed using COMSOL Multiphysics with diluted glycerol as the sample fluid and DI water as sheath flow fluid. The effects of fluid velocities in the channels were studied. In Design III, the overall efficiency is less than that in the first two designs, but the advantage of this design lies in the possibility of a simpler fabrication. Subsequently, parameters such as velocity and viscosity were studied in the case of Design III, and the ideal velocity condition was identified as 150 m/sec for the sample flow, 350µm/sec for the first sheath flow, and 550µm/sec for the second sheath. Simulations were carried out with sample and sheath flow fluids that have different viscosities. It was concluded that the effect of focusing is primarily dependent on the velocity rather than on the viscosity of the fluid

One Step Quick Detection of Cancer Cell Surface Marker by Integrated NiFe-based Magnetic Biosensing Cell Cultural Chip

RGD peptides has been used to detect cell surface integrin and direct clinical effective therapeutic drug selection. Herein we report that a quick one step detection of cell surface marker that was realized by a specially designed NiFe-based magnetic biosensing cell chip combined with functionalized magnetic nanoparticles. Magnetic nanoparticles with 20-30 nm in diameter were prepared by coprecipitation and modified with RGD-4C, and the resultant RGD-functionalized magnetic nanoparticles were used for targeting cancer cells cultured on the NiFe-based magnetic biosensing chip and distinguish the amount of cell surface receptor-integrin. Cell lines such as Calu3, Hela, A549, CaFbr, HEK293 and HUVEC exhibiting different integrin expression were chosen as test samples. Calu3, Hela, HEK293 and HUVEC cells were successfully identified. This approach has advantages in the qualitative screening test. Compared with traditional method, it is fast, sensitive, low cost, easy-operative, and needs very little human intervention. The novel method has great potential in applications such as fast clinical cell surface marker detection, and diagnosis of early cancer, and can be easily extended to other biomedical applications based on molecular recognition

Biocompatible micro-sized cell culture chamber for the detection of nanoparticle-induced IL8 promoter activity on a small cell population

In most conventional in vitro toxicological assays, the response of a complete cell population is averaged, and therefore, single-cell responses are not detectable. Such averaging might result in misinterpretations when only individual cells within a population respond to a certain stimulus. Therefore, there is a need for non-invasive in vitro systems to verify the toxicity of nanoscale materials. In the present study, a micro-sized cell culture chamber with a silicon nitride membrane (0.16 mm2) was produced for cell cultivation and the detection of specific cell responses. The biocompatibility of the microcavity chip (MCC) was verified by studying adipogenic and neuronal differentiation. Thereafter, the suitability of the MCC to study the effects of nanoparticles on a small cell population was determined by using a green fluorescence protein-based reporter cell line. Interleukin-8 promoter (pIL8) induction, a marker of an inflammatory response, was used to monitor immune activation. The validation of the MCC-based method was performed using well-characterized gold and silver nanoparticles. The sensitivity of the new method was verified comparing the quantified pIL8 activation via MCC-based and standard techniques. The results proved the biocompatibility and the sensitivity of the microculture chamber, as well as a high optical quality due to the properties of Si3N4. The MCC-based method is suited for threshold- and time-dependent analysis of nanoparticle-induced IL8 promoter activity. This novel system can give dynamic information at the level of adherent single cells of a small cell population and presents a new non-invasive in vitro test method to assess the toxicity of nanomaterials and other compounds

Microfabrication and Applications of Opto-Microfluidic Sensors

A review of research activities on opto-microfluidic sensors carried out by the research groups in Canada is presented. After a brief introduction of this exciting research field, detailed discussion is focused on different techniques for the fabrication of opto-microfluidic sensors, and various applications of these devices for bioanalysis, chemical detection, and optical measurement. Our current research on femtosecond laser microfabrication of optofluidic devices is introduced and some experimental results are elaborated. The research on opto-microfluidics provides highly sensitive opto-microfluidic sensors for practical applications with significant advantages of portability, efficiency, sensitivity, versatility, and low cost

Particle-Based Microfluidic Device for Providing High Magnetic Field Gradients

A microfluidic device for manipulating particles in a fluid has a device body that defines a main channel therein, in which the main channel has an inlet and an outlet. The device body further defines a particulate diverting channel therein, the particulate diverting channel being in fluid connection with the main channel between the inlet and the outlet of the main channel and having a particulate outlet. The microfluidic device also has a plurality of microparticles arranged proximate or in the main channel between the inlet of the main channel and the fluid connection of the particulate diverting channel to the main channel. The plurality of microparticles each comprises a material in a composition thereof having a magnetic susceptibility suitable to cause concentration of magnetic field lines of an applied magnetic field while in operation. A microfluidic particle-manipulation system has a microfluidic particle-manipulation device and a magnet disposed proximate the microfluidic particle-manipulation device