Brian Bilenberg

Lab-on-a-chip with integrated optical transducers

Taking the next step from individual functional components to higher integrated devices, we present a feasibility study of a lab-on-a-chip system with five different components monolithically integrated on one substrate. These five components represent three main domains of microchip technology: optics, fluidics and electronics. In particular, this device includes an on-chip optically pumped liquid dye laser, waveguides and fluidic channels with passive diffusive mixers, all defined in one layer of SU-8 polymer, as well as embedded photodiodes in the silicon substrate. The dye laser emits light at 576 nm, which is directly coupled into five waveguides that bring the light to five different locations along a fluidic channel for absorbance measurements. The transmitted portion of the light is collected at the other side of this cuvette, again by waveguides, and finally detected by the photodiodes. Electrical read-out is accomplished by integrated metal connectors. To our knowledge, this is the first time that integration of all these components has been demonstrated.

PMMA to SU-8 bonding for polymer based lab-on-a-chip systems with integrated optics

An adhesive bonding technique for wafer-level sealing of SU-8 based lab-on-a-chip microsystems with integrated optical components is presented. Microfluidic channels and optical components, e.g. waveguides, are fabricated in cross-linked SU-8 and sealed with a Pyrex glass substrate by means of an intermediate layer of 950k molecular weight poly-methylmethacrylate (PMMA). Due to a lower refractive index of PMMA (n = 1.49 at λ = 600–900 nm) this bonding technique preserves waveguiding in the cross-linked SU-8 structures (n = 1.59 at λ = 633 nm) in combination with good sealing of the microfluidic channels. The bonding strength dependence on bonding temperature and bonding force is investigated. A maximum bonding strength of 16 MPa is achieved at bonding temperatures between 110 °C and 120 °C, at a bonding force of 2000 N on a 4 inch wafer. The optical propagation loss of multi-mode 10 µm (thickness) × 30 µm (width) SU-8 waveguides is measured. The propagation loss in PMMA bonded waveguide structures is more than 5 dB cm−1 lower, at wavelengths between 600 nm and 900 nm, than in similar structures bonded by an intermediate layer of SU-8. Furthermore 950k PMMA shows no tendency to flow into the bonded structures during bonding because of its high viscosity.

Topas-based lab-on-a-chip microsystems fabricated by thermal nanoimprint lithography

We present a one-step technology for fabrication of Topas-based lab-on-a-chip (LOC) microsystems by the use of thermal nanoimprint lithography (NIL). The technology is demonstrated by the fabrication of two working devices: a particle separator and a LOC with integrated optics for absorbance measurements. These applications demonstrate the fabrication of millimeter to micrometer-sized structures in one lithographic step. The use of NIL makes the technology easily scalable into the nanometer regime by the use of a suitable lithographic technique in the fabrication of the stamp. Processing issues such as environmental stress cracking of the Topas and the requirements to anti-sticking layers on the stamp when imprinting into Topas are discussed.

Imprinted silicon-based nanophotonics

We demonstrate and optically characterize silicon-on-insulator based nanophotonic devices fabricated by nanoimprint lithography. In our demonstration, we have realized ordinary and topology-optimized photonic crystal waveguide structures. The topology-optimized structures require lateral pattern definition on a sub 30-nm scale in combination with a deep vertical silicon etch of the order of ~300 nm. The nanoimprint method offers a cost-efficient parallel fabrication process with state-of-the-art replication fidelity, comparable to direct electron beam writing.

Comparison of high resolution negative electron beam resists

Four high resolution negative electron beam resists are compared: TEBN-1 from Tokuyama Corp. Japan, ma-N 2401XP and mr-L 6000.1XP from microresist technology GmbH Germany, and SU-8 2000 series from MicroChem Corp., USA. Narrow linewidth high density patterns are defined by 100kV electron beam lithography, and the pattern is transferred into silicon by a highly anisotropic SF6∕O2∕CHF3 based reactive ion etch process with a selectivity between silicon and the investigated resists of approximately 2. 20nm half-pitch lines and 10nm lines with a pitch down to 60nm are written and transferred into silicon.

Real-time tunability of chip-based light source enabled by microfluidic mixing

We demonstrate real-time tunability of a chip-based liquid light source enabled by microfluidic mixing. The mixer and light source are fabricated in SU-8 which is suitable for integration in SU-8-based laboratory-on-a-chip microsystems. The tunability of the light source is achieved by changing the concentration of rhodamine 6G dye inside two integrated vertical resonators, since both the refractive index and the gain profile are influenced by the dye concentration. The effect on the refractive index and the gain profile of rhodamine 6G in ethanol is investigated and the continuous tuning of the laser output wavelength is demonstrated using an ethanolic rhodamine 6G solution of 2×10−2mol∕l mixed with pure ethanol. This yields rhodamine 6G concentrations from 5×10−3 to 1.5×10−2mol∕l inside the laser resonators and a wavelength change of 10 nm with a response time of 110 s.

Fully integrated optical systems for lab-on-a-chip applications

The integration of optical transducers is generally considered a key issue in the further development of lab-on-a-chip microsystems. We present a technology for the integration of miniaturized, polymer based lasers, with planar waveguides, microfluidic networks and substrates such as structured silicon. The flexibility of the polymer patterning process, enables fabrication of laser light sources and other optical components such as waveguides, lenses and prisms, in the same lithographic process step on a polymer. The optically functionalised polymer layer can be overlaid on any reasonably flat substrate, such as electrically functionalised Silicon containing photodiodes. This optical and microfluidic overlay, interfaces optically with the substrate through the polymer-substrate contact plane. Two types of integrable laser source devices are demonstrated: microfluidic- and solid polymer dye lasers. Both are based on laser resonators defined solely in the polymer layer. The polymer laser sources are optically pumped with an external laser, and emits light in the chip plane, suitable for coupling into chip waveguides. Integration of the light sources with polymer waveguides, micro-fluidic networks and photodiodes embedded in a Silicon substrate is shown in a device designed for measuring the time resolved absorption of two fluids mixed on-chip. The feasibility of three types of polymers is demonstrated: SU-8, PMMA and a cyclo-olefin co-polymer (COC) -- Topas. SU-8 is a negative tone photoresist, allowing patterning with conventional UV lithography. PMMA and Topas are thermoplasts, which are patterned by nanoimprint lithography (NIL).

Smart plastic functionalization by nanoimprint and injection molding

In this paper, we present a route for making smart functionalized plastic parts by injection molding with sub-micrometer surface structures. The method is based on combining planar processes well known and established within silicon micro and sub-micro fabrication with proven high resolution and high fidelity with truly freeform injection molding inserts. The link between the planar processes and the freeform shaped injection molding inserts is enabled by the use of nanoimprint with flexible molds for the pattern definition combined with unidirectional sputter etching for transferring the pattern. With this approach, we demonstrate the transfer of down to 140 nm wide holes on large areas with good structure fidelity on an injection molding steel insert. The durability of the sub-micrometer structures on the inserts have been investigated by running two production series of 102,000 and 73,000 injection molded parts, respectively, on two different inserts and inspecting the inserts before and after the production series and the molded parts during the production series.

Technology for Fabrication of Nanostructures by Standard Cleanroom Processing and Nanoimprint Lithography

Sub-micron structures are routinely fabricated by electron beam lithography (EBL). However EBL is a time consuming and costly technology. We present a technology for fabrication of nanostructures by standard UV-lithography and thermal nanoimprint lithography (NIL). NIL-stamps with sub-30 nm patterns are fabricated by standard micrometer resolution cleanroom processing, i.e. UV-lithography, reactive ion etching and thermal oxidation, and the pattern is transferred to a polymer thin film on a substrate by NIL. Subsequently the patterned polymer film is used either as a direct etching mask to transfer the pattern to the substrate or as a metal lift-off mask. This way we have demonstrated the fabrication of sub-100 nm nanochannels in silicon oxide and sub-50 nm gold lines on silicon.

Sequencing of human genomes extracted from single cancer cells isolated in a valveless microfluidic device.

Sequencing the genomes of individual cells enables the direct determination of genetic heterogeneity amongst cells within a population. We have developed an injection-moulded valveless microfluidic device in which single cells from colorectal cancer derived cell lines (LS174T, LS180 and RKO) and fresh colorectal tumors have been individually trapped, their genomes extracted and prepared for sequencing using multiple displacement amplification (MDA). Ninety nine percent of the DNA sequences obtained mapped to a reference human genome, indicating that there was effectively no contamination of these samples from non-human sources. In addition, most of the reads are correctly paired, with a low percentage of singletons (0.17 ± 0.06%) and we obtain genome coverages approaching 90%. To achieve this high quality, our device design and process shows that amplification can be conducted in microliter volumes as long as the lysis is in sub-nanoliter volumes. Our data thus demonstrates that high quality whole genome sequencing of single cells can be achieved using a relatively simple, inexpensive and scalable device. Detection of genetic heterogeneity at the single cell level, as we have demonstrated for freshly obtained single cancer cells, could soon become available as a clinical tool to precisely match treatment with the properties of a patients own tumor.