Damla Ozcelik

Optofluidic wavelength division multiplexing for single-virus detection

Significance The ability to simultaneously detect and identify multiple biological particles or biomarkers is one of the key requirements for molecular diagnostic tests that are becoming even more important as personalized and precision medicine place increased emphasis on such capabilities. Integrated optofluidic platforms can help create such highly sensitive, multiplexed assays on a small, dedicated chip. We introduce a method for multiplex fluorescence detection of single bioparticles by creating color-dependent excitation spot patterns from a single integrated waveguide structure. Detection and identification of individual virus particles from three different influenza subtypes are demonstrated. The principle can be readily applied to amplification-free detection of nucleic acid biomarkers as well as larger target numbers using combinatorial color coding. Optical waveguides simultaneously transport light at different colors, forming the basis of fiber-optic telecommunication networks that shuttle data in dozens of spectrally separated channels. Here, we reimagine this wavelength division multiplexing (WDM) paradigm in a novel context––the differentiated detection and identification of single influenza viruses on a chip. We use a single multimode interference (MMI) waveguide to create wavelength-dependent spot patterns across the entire visible spectrum and enable multiplexed single biomolecule detection on an optofluidic chip. Each target is identified by its time-dependent fluorescence signal without the need for spectral demultiplexing upon detection. We demonstrate detection of individual fluorescently labeled virus particles of three influenza A subtypes in two implementations: labeling of each virus using three different colors and two-color combinatorial labeling. By extending combinatorial multiplexing to three or more colors, MMI-based WDM provides the multiplexing power required for differentiated clinical tests and the growing field of personalized medicine.

Integration of programmable microfluidics and on-chip fluorescence detection for biosensing applications

We describe the integration of an actively controlled programmable microfluidic sample processor with on-chip optical fluorescence detection to create a single, hybrid sensor system. An array of lifting gate microvalves (automaton) is fabricated with soft lithography, which is reconfigurably joined to a liquid-core, anti-resonant reflecting optical waveguide (ARROW) silicon chip fabricated with conventional microfabrication. In the automaton, various sample handling steps such as mixing, transporting, splitting, isolating, and storing are achieved rapidly and precisely to detect viral nucleic acid targets, while the optofluidic chip provides single particle detection sensitivity using integrated optics. Specifically, an assay for detection of viral nucleic acid targets is implemented. Labeled target nucleic acids are first captured and isolated on magnetic microbeads in the automaton, followed by optical detection of single beads on the ARROW chip. The combination of automated microfluidic sample preparation and highly sensitive optical detection opens possibilities for portable instruments for point-of-use analysis of minute, low concentration biological samples.

Signal-to-Noise Enhancement in Optical Detection of Single Viruses With Multispot Excitation

We present fluorescence detection of single H1N1 viruses with enhanced signal to noise ratio (SNR) achieved by multispot excitation in liquid-core antiresonant reflecting optical waveguides (ARROWs). Solid-core Y-splitting ARROW waveguides are fabricated orthogonal to the liquid-core section of the chip, creating multiple excitation spots for the analyte. We derive expressions for the SNR increase after signal processing, and analyze its dependence on signal levels and spot number. Very good agreement between theoretical calculations and experimental results is found. SNR enhancements up to 5×104 are demonstrated.

On-chip wavelength multiplexed detection of cancer DNA biomarkers in blood

We have developed an optofluidic analysis system that processes biomolecular samples starting from whole blood and then analyzes and identifies multiple targets on a silicon-based molecular detection platform. We demonstrate blood filtration, sample extraction, target enrichment, and fluorescent labeling using programmable microfluidic circuits. We detect and identify multiple targets using a spectral multiplexing technique based on wavelength-dependent multi-spot excitation on an antiresonant reflecting optical waveguide chip. Specifically, we extract two types of melanoma biomarkers, mutated cell-free nucleic acids -BRAFV600E and NRAS, from whole blood. We detect and identify these two targets simultaneously using the spectral multiplexing approach with up to a 96% success rate. These results point the way toward a full front-to-back chip-based optofluidic compact system for high-performance analysis of complex biological samples.

Spectrally reconfigurable integrated multi-spot particle trap

Optical manipulation of small particles in the form of trapping, pushing, or sorting has developed into a vast field with applications in the life sciences, biophysics, and atomic physics. Recently, there has been increasing effort toward integration of particle manipulation techniques with integrated photonic structures on self-contained optofluidic chips. Here, we use the wavelength dependence of multi-spot pattern formation in multimode interference (MMI) waveguides to create a new type of reconfigurable, integrated optical particle trap. Interfering lateral MMI modes create multiple trapping spots in an intersecting fluidic channel. The number of trapping spots can be dynamically controlled by altering the trapping wavelength. This novel, spectral reconfigurability is utilized to deterministically move single and multiple particles between different trapping locations along the channel. This fully integrated multi-particle trap can form the basis of high throughput biophotonic assays on a chip.

Tailoring the spectral response of liquid waveguide diagnostic platforms

Liquid filled waveguides that form the basis for on-chip biophotonics diagnostic platforms have primarily found application in fluorescence and Raman spectroscopy experiments that require sensitive discrimination between weak analyte signals and a variety of background signals. Primary sources of background signal can include light from excitation sources (strong, narrow frequency band) and photoluminescence generated in waveguide cladding layers (weak, wide frequency band). Here we review both solid and liquid core filtering structures which are based on anti-resonant reflection that can be integrated with waveguides for attenuating undesirable optical bands. Important criteria to consider for an optimized biosensor include cladding layer materials that minimize broad-spectrum photoluminescence and optimize layer thicknesses for creating a desired spectral response in both solid and liquid guiding layers, and a microfabrication process capable of producing regions with variable spectral response. New results describing how spurious fluorescence can be minimized by optimized thermal growth conditions and how liquid-core filter discrimination can be tuned with liquid core waveguide length are presented.

Improved environmental stability for plasma enhanced chemical vapor deposition SiO2 waveguides using buried channel designs

Ridge and buried channel waveguides (BCWs) made using plasma-enhanced chemical vapor deposition SiO2 were fabricated and tested after being subjected to long 85°C water baths. The water bath was used to investigate the effects of any water absorption in the ridge and BCWs. Optical mode spreading and power throughput were measured over a period of three weeks. The ridge waveguides quickly absorbed water within the critical guiding portion of the waveguide. This caused a nonuniformity in the refractive index profile, leading to poor modal confinement after only seven days. The BCWs possessed a low index top cladding layer of SiO2, which caused an increase in the longevity of the waveguides, and after 21 days, the BCW samples still maintained ~20% throughput, much higher than the ridge waveguides, which had a throughput under 5%.

Scalable Spatial-Spectral Multiplexing of Single-Virus Detection Using Multimode Interference Waveguides

Simultaneous detection of multiple pathogens and samples (multiplexing) is one of the key requirements for diagnostic tests in order to enable fast, accurate and differentiated diagnoses. Here, we introduce a novel, highly scalable, photonic approach to multiplex analysis with single virus sensitivity. A solid-core multimode interference (MMI) waveguide crosses multiple fluidic waveguide channels on an optofluidic chip to create multi-spot excitation patterns that depend on both the wavelength and location of the channel along the length of the MMI waveguide. In this way, joint spectral and spatial multiplexing is implemented that encodes both spatial and spectral information in the time dependent fluorescence signal. We demonstrate this principle by using two excitation wavelengths and three fluidic channels to implement a 6x multiplex assay with single virus sensitivity. High fidelity detection and identification of six different viruses from a standard influenza panel is reported. This multimodal multiplexing strategy scales favorably to large numbers of targets or large numbers of clinical samples. Further, since single particles are detected unbound in flow, the technique can be broadly applied to direct detection of any fluorescent target, including nucleic acids and proteins.

Silicate overcoat layers for the improvement of PECVD SiO 2 optofluidic waveguides

Silicate spin-on-glass can be used as a protective barrier to coat PECVD SiO2 optofluidic waveguides in order to smooth out surface topology, and lessen moisture absorption on top of the waveguide. The measured optical throughput and mode confinement improved when compared to uncoated waveguides.

Optofluidic bioanalysis: fundamentals and applications

Abstract: Over the past decade, optofluidics has established itself as a new and dynamic research field for exciting developments at the interface of photonics, microfluidics, and the life sciences. The strong desire for developing miniaturized bioanalytic devices and instruments, in particular, has led to novel and powerful approaches to integrating optical elements and biological fluids on the same chip-scale system. Here, we review the state-of-the-art in optofluidic research with emphasis on applications in bioanalysis and a focus on waveguide-based approaches that represent the most advanced level of integration between optics and fluidics. We discuss recent work in photonically reconfigurable devices and various application areas. We show how optofluidic approaches have been pushing the performance limits in bioanalysis, e.g. in terms of sensitivity and portability, satisfying many of the key requirements for point-of-care devices. This illustrates how the requirements for bianalysis instruments are increasingly being met by the symbiotic integration of novel photonic capabilities in a miniaturized system.