This PDF file contains the front matter associated with SPIE Proceedings Volume 9725 including the Title Page, Copyright information, Table of Contents, Introduction, and Conference Committee listing.

The interaction of light with biological cells can provide a unique tool for studying their biophysical properties. Optical spectroscopy of biological cells can reveal detailed information on their structure and dynamics in a way that is not possible with traditional microscopy techniques. Histological evaluation can only obtain a snapshot of the activity of individual cells, relying instead on large ensembles to develop a picture of their temporal evolution. On the other hand optical spectroscopy can be applied to cells with little to no preparation and can enable studies of the same live cells at extended time intervals. Our research group has developed a suite of optical spectroscopic tools to assess the structure and function of biological cells and modulation due to the onset of disease. The wavelength dependence of the interaction of cells with light provides information of cell features through elastic scattering across the visible and near infrared spectrum. Alternatively, the angular dependence of scattered light can also be used to reveal cell properties. We will discuss how both modes of elastic scattering can be used to evaluate cell status. Finally, the recent advances in the use of optical phase imaging to create contrast in nearly transparent biological cells will also be discussed as related to the role of this modality in biosensing.

Recently exosomes have attracted interest due to their potential as cancer biomarkers. We report the real time, label‐free sensing of single exosomes in serum using microtoroid optical resonators. We use this approach to assay the progression of tumors implanted in mice by specifically detecting low concentrations of tumor‐derived exosomes. Our approach measures the adsorption of individual exosomes onto a functionalized silica microtoroid by tracking changes in the optical resonant frequency of the microtoroid. When exosomes land on the microtoroid, they perturb its refractive index in the evanescent field and thus shift its resonance frequency. Through digital frequency locking, we are able to rapidly track these shifts with accuracies of better than 10 attometers (one part in 10^11). Samples taken from tumor‐implanted mice from later weeks generated larger frequency shifts than those from earlier weeks. Control samples taken from a mouse with no tumor generated no such increase in signal between subsequent weeks. Analysis of shifts from tumor-implanted mouse samples show a distribution of unitary steps, with the maximum step having a height of ~1.2 fm, corresponding to an exosome size of 44 ± 4.8 nm. This size range corresponds to that found by performing nanoparticle tracking analysis on the same samples. Our results demonstrate development towards a minimally‐invasive tumor "biopsy" that eliminates the need to find and access a tumor.

We have demonstrated a Cu2O/ZnO nanorods (NRs) array p-n heterostructures photoelectrochemical biosensor. The electrodeposition of Cu2O at pH 12 acquired the preferably (111) lattice planes, resulting in the largest interfacial electric field between Cu2O and ZnO, which finally led to the highest separation efficiency of photogenerated charge carriers. High verticality ZnO nanorods by seed layer and thermal annealing assist the hydrothermal growth. The optimized Cu2O/ZnO NRs array p-n heterostructures exhibited enhanced PEC performance, such as elevated photocurrent and photoconversion efficiency, as well as excellent sensing performance for the sensitive detection of four strains of different races and different degree of cancer cell which made the device self-powered. We got spectral response characteristics and operating wavelength range of biosensor, and to verify the biological characteristics of cancer cells wafer react with different stages of cancer characterized by a cancer measured reaction experiment.

Biodynamic imaging is an emerging 3D optical imaging technology that probes up to 1 mm deep inside three-dimensional living tissue using short-coherence dynamic light scattering to measure the intracellular motions of cells inside their natural microenvironments. Biodynamic imaging is label-free and non-invasive. The information content of biodynamic imaging is captured through tissue dynamics spectroscopy that displays the changes in the Doppler signatures from intracellular constituents in response to applied compounds. The affected dynamic intracellular mechanisms include organelle transport, membrane undulations, cytoskeletal restructuring, strain at cellular adhesions, cytokinesis, mitosis, exo- and endo-cytosis among others. The development of 3D high-content assays such as biodynamic profiling can become a critical new tool for assessing efficacy of drugs and the suitability of specific types of tissue growth for drug discovery and development. The use of biodynamic profiling to predict clinical outcome of living biopsies to cancer therapeutics can be developed into a phenotypic companion diagnostic, as well as a new tool for therapy selection in personalized medicine. This invited talk will present an overview of the optical, physical and physiological processes involved in biodynamic imaging. Several different biodynamic imaging modalities include motility contrast imaging (MCI), tissue-dynamics spectroscopy (TDS) and tissue-dynamics imaging (TDI). A wide range of potential applications will be described that include process monitoring for 3D tissue culture, drug discovery and development, cancer therapy selection, embryo assessment for in-vitro fertilization and artificial reproductive technologies, among others.

We used silicon nitride (Si3N4) directional coupler (DC) interferometers to fabricate evanescent field biosensors for detection of aptamer hybridization. The DC detects temporal changes in refractive index (RI) as changes in the relative intensities of near-fields. Numerical simulation provided information for sensitivity of the DC as well as preferable dimensions of single mode waveguides. The result suggests that lengthening the DC improves its sensitivity. Another approach to realize high sensitivity was to make the coupling region of the DC longer for a given complete coupling length L_c. A folded DC is also presented based on calculations using Si waveguides. Here a meander-line with an array of segmented DCs and asymmetric 180°-curved waveguides was accommodated in a (100 μm)² square area. Phases of propagating lights within two curved waveguides could be coincided. This cascade-connection of the segments of the DCs facilitated highly sensitive detection of biomolecules and a flexible sensor design. Signal dependences on device length and bulk RI change were evaluated by end-fire coupling measurements. The optical intensities agreed well with the corresponding fitting curve and the curve calculated based on the coupled mode theory.

Heavy metal ions released into the environment from industrial processes lead to various health hazards. We propose an on-chip label-free detection approach that allows high-sensitivity and high-throughput detection of heavy metals. The sensing device consists of 2-dimensional photonic crystal microcavities that are combined by multimode interferometer to form a sensor array. We experimentally demonstrate the detection of cadmium-chelate conjugate with concentration as low as 5 parts-per-billion (ppb).

Clinical attrition is about 30% from failure of drug candidates due to toxic side effects, increasing the drug development costs significantly and slowing down the drug discovery process. This partly originates from the fact that the animal models do not accurately represent human physiology. Hence there is a clear unmet need for developing drug toxicity assays using human-based models that are complementary to traditional animal models before starting expensive clinical trials. Organ-on-a-chip techniques developed in recent years have generated a variety of human organ models mimicking different human physiological conditions. However, it is extremely challenging to monitor the transient and long-term response of the organ models to drug treatments during drug toxicity tests. First, when an organ-on-a-chip model interacts with drugs, a certain amount of protein molecules may be released into the medium due to certain drug effects, but the amount of the protein molecules is limited, since the organ tissue grown inside microfluidic bioreactors have minimum volume. Second, traditional fluorescence techniques cannot be utilized for real-time monitoring of the concentration of the protein molecules, because the protein molecules are continuously secreted from the tissue and it is practically impossible to achieve fluorescence labeling in the dynamically changing environment. Therefore, direct measurements of the secreted protein molecules with a label-free approach is strongly desired for organs-on-a-chip applications. In this paper, we report the development of a photonic crystal-based biosensor for label-free assays of secreted protein molecules from a liver-on-a-chip model. Ultrahigh detection sensitivity and specificity have been demonstrated.

We report an optical cavity based biosensor using a novel differential detection method for point-of-care applications. Two laser diodes allow for multiplexing capability along with the ability to enhance the responsivity using differential detection. The laser wavelengths are chosen so that the optical intensities of two lasers change monotonically with opposite slopes upon the adsorption of desired biomarkers. The cavity width, PMMA thickness, and silver thickness have been optimized to achieve a large change in scaled differential value. We chose biotinylated BSA detection with Avidin as a receptor molecule to demonstrate the proposed design. Avidin is attached directly to the PMMA layer by physisorption. Then, biotinylated BSA is introduced to the sample and the intensities of the laser diodes are measured by a sCMOS camera. A change in the scaled differential value will correlate to the binding of biotinylated BSA. In this presentation, we will discuss simulation results, fabrication procedures, and preliminary measurement results.

Optical biosensors have emerged in the past decade as the most promising candidates for portable, highly-sensitive bioanalytical systems that can be employed for in-situ measurements. In this work, a miniaturized optoelectronic system for rapid, quantitative, label-free detection of harmful species in food is presented. The proposed system has four distinctive features that can render to a powerful tool for the next generation of Point-of-Need applications, namely it accommodates the light sources and ten interferometric biosensors on a single silicon chip of a less-than-40mm2 footprint, each sensor can be individually functionalized for a specific target analyte, the encapsulation can be performed at the wafer-scale, and finally it exploits a new operation principle, Broad-band Mach-Zehnder Interferometry to ameliorate its analytical capabilities. Multi-analyte evaluation schemes for the simultaneous detection of harmful contaminants, such as mycotoxins, allergens and pesticides, proved that the proposed system is capable of detecting within short time these substances at concentrations below the limits imposed by regulatory authorities, rendering it to a novel tool for the near-future food safety applications.

Colorimetric detection is cost-effective and user-friendly when used for sensing target analytes without a need of bulky and expensive equipment. The extraordinary transmission phenomena through plasmonic periodic nanocup arrays achieve colorimetric sensing by detecting color changes of transmitted light associated with the refractive index variation. The application of the nanocup arrays, however, is relatively restricted due to a limited sensitivity for monolayered target analyte detections on the surface. In order to improve the sensitivity bounded by the underlying nanostructures, hybrid nanoparticle (NP) – nanocup array substrates are developed for enhancing the sensitivity to the refractive index change. The three dimensionally assembled Au NPs in circle along the sidewall of each nanocup increases the density of hot spots by the heterogeneous plasmonic coupling between the NP and the edge of the nanocup; thus a small refractive index change at the hot spot becomes easily detected than bare nanocup arrays. In addition to the bulk refractive index sensing, an ultrasensitive spectroscopic detection of the antigen-antibody binding is achieved by this three-dimensional self-assembly of Au NPs on the Au nanocup arrays.

Plasmonic biosensors hold enormous potential for the development of low-cost, label-free, point-of-care biodiagnostics. However, two major challenges need to be overcome to reap the benefits of this class of biosensors: (i) state-of-the-art plasmonic biosensors either offer limited sensitivity or are impractical for real-world applications due to their poor stability and excessive cost; (ii) these biosensors rely on natural antibodies, which are high-cost and impose severe limitations in handling, storage and device integration. We demonstrate that a common filter paper can be transformed into a plasmonic sensing platform for highly sensitive and selective detection of trace levels of chemical and biological analytes. We also demonstrate that short peptides as biorecognition elements compared to larger antibodies as target capture agents offer several advantages. Using a bioplasmonic paper device, we demonstrate the selective and sensitive detection of the cardiac biomarker troponin I. The smaller sized peptide provides higher sensitivity and a lower detection limit using a bioplasmonic paper device. Furthermore, the excellent shelf-life and thermal stability of peptide-based plasmonic biosensors, which precludes the need for special storage conditions, makes it ideal for use in resource-limited settings. We also demonstrate plasmonic biosensors based on artificial antibodies by molecularly imprinting the plasmonic nanotransducers. Apart from significantly lowering the cost, these developments are critical for translating plasmonic sensors to point-of-care and resource-limited settings.

G-quadruplex, readily formed by the G-rich sequence, potentially distributes in over 40 % of all human genes, such as the telomeric DNA with the G-rich sequence found at the end of the chromosome. The G-quadruplex structure is supposed to possess a diverse set of critical functions in the mammalian genome for transcriptional regulation, DNA replication and genome stability. However, most of the currently available methods for G-quadruplex identification are restricted to fluorescence techniques susceptible to poor sensitivity. It is essential to propose methods with higher sensitivity to specifically recognize the G-quadruplexes. In this study, we demonstrate a label-free plasmonic biosensor for G-quadruplex detection by relying on the advantages of nanoporous gold (NPG) disks that provide high-density plasmonic hot spots, suitable for molecular recognition capability without the requirement for labeling processes.

Plasmonic biosensors have greatly overcome the limitations of conventional optical sensors in terms of sensitivity, tunability, photo-stability, and in vivo applicability. In this paper, we present plasmonic biosensors using bioenabled nanomaterials diatom biosilica, with active surface functionalities as affordable and eco-friendly integration platforms of Ag nanoparticles for label-free detection of biomolecules. Dual-mode plasmon sensing mechanisms, including surface-enhanced Raman scattering (SERS) and refractive-index (RI) sensing will be simultaneously implemented on the plasmonic-biosilica nanostructures to obtain quantitative biosensing with structural resolution of the biomolecules. We have achieved ultra-sensitive detection of Rhodamine 6G (R6G) at concentrations as low as 10⁻¹⁰ M. Furthermore, this substrate was used to detect TNT, illustrating the potential application as viable substrates for monitoring pollutant and toxics in environment.

Direct measurements of the strength of particle interactions are critical for characterizing the stability and behavior of colloidal and nanoparticle suspensions. Current techniques are limited in their ability to measure pico-newton scale interaction forces on sub-micrometer particles due to signal detection limits, thermal noise, and throughput. We have recently developed a technique for making direct mechanical measurements of the force and work associated with the steric and electrostatic effects that stabilize colloidal nanoparticles. “Nanophotonic Force Microscopy”, as we call it, is unique in that it uses statistical methods to provide direct measurements of these forces at the individual particle scale, while still being sufficiently high-throughput to produce meaningful population level data. In this talk I will introduce the technology, it’s advantages, and some of the major uses. Specific case studies will include label-free monitoring of binding of individual antibodies onto single viruses and the measurement of the strength of nanoparticle coatings used for steric stabilization.

The integration of optofluidic laser and FRET mechanism provides novel research frontiers, including sensitive biochemical analysis and novel photonic devices, such as on-chip coherent light sources and bio-tunable lasers. Here we investigated an optofluidic FRET laser using quantum dots (QDs) as FRET donors. We achieved lasing from Cy5 as the acceptor in the QD-Cy5 pair with excitation at 450 nm where Cy5 has negligible absorption by itself. The threshold was approximately 14 μJ/mm². The demonstrated capability of QDs as the donor in a FRET laser greatly improves the versatility of optofluidic laser operation due to the broad and large absorption cross section of QDs in the blue and UV spectral region. The excitation efficiency of the acceptor molecules through FRET channel was also analyzed, showing that the energy transfer rate and the non-radiative Auger recombination rate of QDs plays a significant role in FRET laser performance.

The optofluidic laser is an emerging technology that integrates microfluidics, miniaturized laser cavity, and laser gain medium in liquid. It is unique due to its biocompatibility, thus can be used for unconventional bioanalysis, in which biointeraction or process takes place within the optical cavity mode volume. Rather than using fluorescence, the optofluidic laser based detection employs laser emission, i.e., stimulated emission, as the sensing signal, which takes advantage of optical amplification provided by the laser cavity to achieve much higher sensitivity. In this presentation, I will first introduce the concept of optofluidic laser based bioanalysis. Then I will discuss each of the three components (cavity, gain medium, and fluidics) of the optofluidic laser and describe how to use the optofluidic laser in bioanalysis at the molecular, cellular, and tissue level. Finally, I will discuss future research and application directions.

We have shown recently that the hybridization of short oligonucleotides can be studied in a zero-mode waveguide nanostructure (ZMW) chip using a modified DNA sequencer.[1] Here we present an extension of this method enabling the parallel measurement of kinetic constants of a large number of hybridization reactions on a single chip. This can be achieved by immobilization of a mixture of oligonucleotides, which leads to a statistical and random distribution of single molecules in the 150’000 ZMWs of a SMRT™ cell. This setup is comparable to a classical microarray with ZMWs in place of spots but unknown allocation of probes. The probe surface density is reduced by a factor of ~10¹⁰ allowing the study of hybridization in the absence of interactions with neighboring probes. Hybridization with a dye labelled oligonucleotide results in trains of fluorescence pulses from which interpulse durations (IPDs) and pulse widths (PWs) can be extracted. Since the identity of a probe in a ZMW is unknown, the immobilized oligonucleotide is sequenced in a subsequent step. After mapping the fluorescence traces to the sequence, the association and dissociation rate constant for each oligonucleotide can be calculated. By selecting suitable probes, the method can be used to determine rate constants of hybridization for a large number of mismatch oligonucleotides in a single measurement and at single-molecule level.

Waste water, drinking water and other industrial water sources are more and more/increasingly polluted with a large variety of contaminants, such as pesticides or residuals of pharmaceuticals. These compounds can impact human and animal organisms and lead to serious health issues. Today, in order to analyze the presence and quantity of the abovementioned micropollutants, samples are typically sent to specialized centralized laboratories and their processing may take up to several days. In order to meet the demand for continuous and consistent monitoring of aqueous solutions we propose a novel label-free technology system comprising proprietary chip and reader device designs. The core of the system is constituted by a planar-grated-waveguide (PGW) chip. Label-free biosensors, based on PGWs are sensitive to effective refractive index changes caused by the adsorption of biomolecules (micropollutants) onto the sensor surface or due to refractive index changes of the bulk solution. The presented reader device operates with a novel readout concept based on a scanning MEMS mirror for the angular interrogation of input grating couplers at a high repetition rate. The reader has fully integrated optics, electronics and fluidics and at the same time consumes limited energy (portable, field use ready). In the recent experiments, the effectiveness of the technology has been demonstrated with various liquids and bioassays showing (i) an excellent refractometric sensitivity with a limit of detection towards effective refractive index changes of ▵neff < 2 x 10-7, and (ii) the capability to perform affinity measurements for large (<150 kDa) and small (<250 Da) molecules.

We report on a silicon-photonic cascaded microring sensor with thermo-optic tuning for extension of detection range. Cascaded microring resonators have been used to realize refractive index sensors with high sensitivity in the intensity detection mode. In this configuration, one ring is used to probe analyte while the other ring (called filter) is used for spectrum overlap measurement. A significant drawback of this configuration is decreased range of detection. We overcome this problem by thermo-optically tuning the spectrum the filter ring to track the position of maximum overlap, as the spectrum of the probing ring undergoes shift. Using this peak tracking method, we have experimentally demonstrated range enhancement by a factor of 7.8, compared to the intensity detection scheme. This method can use a broadband source for operation, and has a potential for development of low cost, point of care biomedical applications.

Label-free biosensors promised a paradigm involving direct detection of biomarkers from complex samples such as serum without requiring multistep sample processing typical of labelled methods such as ELISA or immunofluorescence assays. Label-free sensors have witnessed decades of development with a veritable zoo of techniques available today exploiting a multitude of physical effects. It is appropriate now to critically assess whether label-free technologies have succeeded in delivering their promise with respect to diagnostic applications, particularly, ambitious goals such as early cancer detection using serum biomarkers, which require low limits of detection (LoD). Comparison of nearly 120 limits of detection (LoD) values reported by labelled and label-free sensing approaches over a wide range of detection techniques and target molecules in serum revealed that labeled techniques achieve 2-3 orders of magnitude better LoDs. Data from experiments where labelled and label-free assays were performed simultaneously using the same assay parameters also confirm that the LoD achieved by labelled techniques is 2 to 3 orders of magnitude better than that by label-free techniques. Furthermore, label-free techniques required significant signal amplification, for e.g. using nanoparticle conjugated secondary antibodies, to achieve LoDs comparable to labelled methods substantially deviating from the original “direct detection” paradigm. This finding has important implications on the practical limits of applying label-free detection methods for molecular diagnosis.

We engineered an effective GO/Ag substrate with a highly controlled and uniform bilayer structure. The substrate was obtained by fine-tuning in the Ag nanocubes (AgNCs) self-assembly, followed by a controlled adsorption of GO sheets on the silver layer. Here the GO coating supplies the signal with additional amplification. The developed GO/AgNCs assemblies show a highly uniform signal distribution, which is ascribed to a homogeneous and soft arrangement of GO sheets over the AgNCs surface. The above characteristics make the GO/AgNCs substrate a reliable tool for detecting different compounds ranging from small molecules to complex biomolecules.