This PDF file contains the front matter associated with SPIE Proceedings Volume 8099, including the Title Page, Copyright information, Table of Contents, and the Conference Committee listing.

The most concerned factors for cancer prognosis are tumor invasion and metastasis. The patterns of tumor invasion can be characterized as random infiltration to surrounding extracellular matrix (ECM) or formation of long-range path for collective migration. Recent studies indicate that mechanical force plays an important role in tumor infiltration and collective migration. However, how tumor colonies develop mechanical interactions with each other to initiate various invasion patterns is unclear. Using a micro-patterning technique, we partition cells into clusters to mimic tumor colonies and quantitatively induce colony-ECM interactions. We find that pre-malignant epithelial cells, in response to concentrations of type I collagen in ECM ([COL]), develop various branching patterns resembling those observed in tumor invasion. In contrast with conventional thought, these patterns require long-range (~ 600 μm) transmission of traction force, but not biochemical factors. At low [COL], cell colonies synergistically develop pairwise and directed branching mimicking the formation of long-range path. By contrast, at high [COL] or high colony density, cell colonies develop random branching and scattering patterns independent of each other. Our results suggest that tumor colonies might select different invasive patterns depending on their interactions with each other and with the ECM.

We face the challenge of providing adequate medical attention to a growing and aging population. Even societies with the best healthcare standards are not prepared to provide adequate medical attention to a growing population. Globally, these problems are magnified as medical care is a mélange ranging from obsolete techniques to state-of-the-art care. A solution to providing proper healthcare in every society, and closing the gap between developed and underserved communities, is the implementation of wireless based preventive medicine. The key components to universalize wireless health care are device miniaturization, increased shelf-life of bio-reagents, and low production cost of medical devices.

We report a portable, low-cost, and high-performance microfluidics based fluorescence-activated cell sorter (microFACS) system to isolate E.coli. cells in combination with a modified specific fluorescence labeling method called tyramide signal amplification-fluorescence in situ hybridization (TSA-FISH). One of the primary challenges in studying bacterial communities that elude cell culturing is to isolate of low abundance bacteria cell from heterogeneous microbial samples. The proposed TSA-FISH protocol is flow cytometry compatible and yields about 10-fold enhancement in fluorescence labeling intensity over widely used standard FISH staining methods. Teflon AF coated optofluidic waveguide and space-time coding with a matched filter algorithm enhance its detection sensitivity. The microFACS is also able to enrich TSA-FISH labeled E.coli. cells by a factor of 223 with an integrated piezoelectric actuator and realtime control electronics system. The microFACS in conjunction with the modified TSA-FISH technologies demonstrates a highly effective and low cost solution potentially for the genomic complexity of complex bacterial communities.

Small interfering RNA (siRNA) is potentially a promising tool in influencing gene expression with a high degree of target specificity. However, its poor intracellular uptake, instability in vivo, and non-specific immune stimulations impeded its effect in clinical applications. In this study, carbon nanotubes (CNTs) functionalized with two types of phospholipid-polyethylene glycol (PEG) have shown capabilities to stabilize siRNA in cell culture medium during the transfection and efficiently deliver siRNA into neuroblastoma and breast cancer cells. Moreover, the intrinsic optical properties of CNTs have been investigated through absorption and fluorescence measurements. We have found that the directly-functionalized groups play an important role on the fluorescence imaging of functionalized CNTs. The unique fluorescence imaging and high delivery efficiency make CNTs a promising material to deliver drugs and evaluate the treatment effect simultaneously.

Although real-time PCR (RT-PCR) has become a diagnostic standard for rapid identification of bacterial species, typical methods remain time-intensive due to sample preparation and amplification cycle times. The assay described in this work incorporates on-chip dielectrophoretic capture and concentration of bacterial cells, thermal lysis, cell permeabilization, and nucleic acid denaturation and fluorescence resonance energy transfer assisted in-situ hybridization (FRET-ISH) species identification. Identification is achieved completely on chip in less than thirty minutes from receipt of sample compared to multiple hours required by traditional RT-PCR and its requisite sample preparation.

This study describes the preparation of hydroxyapatite microspheres for local drugs delivery. The formation of the hydroxyapatite microspheres was initiated by enzymatic decomposition of urea and accomplished by emulsification process (water-in-oil). The microspheres obtained were sintered at 500°C. Scanning electron microscope (SEM) indicated that the microspheres have various porous with random size, which maximizes the surface area. Cytotoxicity was not observed after sintering. Osteoporosis drugs, alendronate and BMP-2, were loaded into HAp microspheres and the releases of both molecules showed sustained releasing profiles.

This work reports the immobilization of monomeric, dimeric and trimer protein Gs onto silica magnetic nanoparticles for self-oriented antibody immobilization. To achieve this, we initially prepared the silica-coated magnetic nanoparticle having about 170 nm diameters. The surface of the silica coated magnetic nanoparticles was modified with 3- aminopropyl-trimethoxysilane (APTMS) to chemically link to multimeric protein Gs. The conjugation of amino groups on the SiO2-MNPs to cysteine tagged in multimeric protein Gs was performed using a sulfo-SMCC coupling procedure. The binding efficiencies of monomer, dimer and trimer were 77 %, 67 % and 55 % respectively. However, the efficiencies of antibody immobilization were 70 %, 83 % and 95 % for monomeric, dimeric and trimeric protein G, respectively. To prove the enhancement of accessibility by using multimeric protein G, FITC labeled goat-anti-mouse IgG was treated to mouse IgG immobilized magnetic silica nanoparticles through multimeric protein G. FITC labeled goat anti-mouse IgGs were more easily bound to mouse IgG immobilized by trimeric protein G than others. Finally protein G bound silica magnetic nanoparticles were utilized to develop highly sensitive immunoassay to detect hepatitis B antigen.

High-sensitivity, label-free biosensors, such as optical microcavities, have shown tremendous potential in medical diagnostics, environmental monitoring, and food safety evaluation, particularly when paired with a biochemical recognition element that grants high specificity towards a target of interest. Their primary limitation is that these systems are single-use, unless the recognition element can be regenerated. Therefore, the ability to selectively functionalize the optical microcavity for a specific target molecule and then recycle the system, without degrading device performance, is extremely important. Here, we present a bioconjugation strategy that not only imparts specificity to optical microcavities, but also allows for biosensor recycling. In this approach, we selectively functionalize the surface of silica microtoroids with a biotin recognition element. We then use a non-destructive O2 plasma treatment to remove the surface chemistry, refresh the recognition element, and recycle the device. The surface chemistry and optical performance of the functionalized and recycled devices are characterized by microcavity analysis, and typical spectroscopic techniques, respectively. The resulting devices can be recycled several times without performance degradation, and show high density surface coverage of biologically active recognition elements. This work represents one of the first examples of a recyclable, bioconjugation strategy for optical microtoroid resonators.

The development of a sensing platform capable of detecting and identifying hazards including biological, chemical, and energetic in nature is a long sought after goal of the Army and many other first responders. Surface enhanced Raman scatting (SERS) is one spectroscopic technique gaining popularity as a solution to many sensing needs due to its many advantages such as high sensitivity, little to no sample preparation required, and use in numerous environmental settings). Despite all the advantages of SERS, it still remains a marginalized sensing technique primarily due to the challenges in fabricating a reliable, highly sensitive and reproducible nanoscale surface. In this work, we show that many of these challenges have been overcome with a newly developed commercially available Klarite SERS substrate. These substrates are fabricated in a fashion similar to standard Klarite substrates, but due to changes in size and spacing of the inverted pyramidal structurethere is an overall increase of SERS sensing capabilities of up to 4 orders of magnitude. In this proceeding paper, the next generation Klarite (308 and 309) substrates are characterized, analyte sensitivity demonstrated at 633 nm and 785 nm, and a brief discussion of their biological sensing capabilities is presented.

Measuring the binding kinetics of molecular systems is fundamental in understanding the interaction between biomolecules within a binding pair. One emerging label-free detection method is based on silica optical microcavities. The majority of research to date with microcavity-based sensors has focused on applications in the diagnostics realm. Here, we develop and characterize a covalent surface attachment strategy for microsphere resonators. We also measure the optical performance (quality factor) of the functionalized microcavities and use them to determine the dissociation constant of the biotinstreptavidin pair. The measured value is within acceptable range of previously published dissociation constants for the biotin-streptavidin pair.

An optical label free and high sensitivity plasmonic biosensor using nanoimprint metallic binary grating is presented based on the phase information of the ellipsometry signal. Plasmonic binary grating was prepared by using soft nanoimprinting technique which significantly reduce the fabrication cost and can be realized for a transition from a laboratory-scale method to full-scale technology. The bulk sensitivity measurement from this 1D binary metallic grating gives a value of refractive index resolution of 1.06×10^-7 RIU. Such a highly sensitive plasmonic biochip was used to investigate the adsorption of bio-molecules on the nanostructure surface in dynamic mode by monitoring the change in polarization state or phase of reflected light in the ellipsometry measurement as a sensing signal.

The paper presents the brief review of published results as well as the original study of photoluminescence (PL) and Raman scattering of core-shell CdSe/ZnS quantum dots (QDs) with radiative interface states. First commercially available CdSe/ZnS QDs with emission at 525 nm (2.36 eV), 565 nm (2.20 eV), 605 nm (2.05 eV) and 640 nm (1.96 eV) covered by PEG polymer have been compared in nonconjugated states. PL spectra of nonconjugated QDs are characterized by a superposition of PL bands related to exciton emission in CdSe cores and to hot electron-hole emission via high energy states (2.00, 2.20, 2.37, 2.75 and 3.04 eV). The high energy states were studded using QDs of different sizes and at different temperatures. It is shown that these PL bands related to interface states. Then the CdSe/ZnS QDs with the color emission 525nm and 605 nm have been conjugated with bio-molecules - ovarian cancer (OC 125) and anti Interleukin 10 (IL-10) antibodies, respectively. It is revealed that the PL spectrum of bioconjugated QDs has changed dramatically with essential decreasing the hot electron-hole recombination flow via interface states. The variation of PL spectra at the bioconjugation is explained on the base of electrostatic interaction and re-charging of QD interface states. The Raman scattering study of nonconjugated and bioconjugated QDs has shown that mentioned antibodies are characterized by the dipole moment that provokes the surface enhance Raman scattering effect in bioconjugated QD samples as well.

Recently, cheap silicon-on-insulator label-free ring resonator biosensors have been demonstrated that allow fast and accurate quantitative detection of biologically relevant molecules for applications in medical diagnostics and drug development. However, a further improvement of their detection limit is limited by their small sensitivity and an expensive tunable laser is typically required to resolve the sharp resonances for wavelength interrogation. Therefore, we experimentally investigated the use of a Vernier-cascade sensor that achieves a sensitivity (thousands of nm/RIU) that is an order of magnitude larger than that of a ring resonator sensor (approx. 100 nm/RIU), while still maintaining sharp spectral features that allow precise monitoring of spectral shifts with data-fitting. Moreover we prove that it's also possible to accurately interrogate the sensor with a low-cost broadband light source by integrating it with an arrayed waveguide grating spectral filter that divides the sensor's transmission spectrum in multiple wavelength channels and transmits them to spatially separated output ports. Experiments show that this sensor can monitor refractive index changes of watery solutions in real-time with a detection limit (1.6 • 10^-5 RIU) competitive with more expensive interrogation schemes, indicating its applicability in low-cost label-free biosensing.

We have created an enhanced cell-imaging platform for 3D confocal fluorescence cell imaging where fluorescence sensitivity is amplified for more than 100 folds especially for cell membrane and cytoplasm. The observed unprecedented three-dimensional fluorescence intensity enhancement on the entire cell microstructure including cell membrane 10 μm above the substrate surface is attributed to a novel far field enhancement mechanism, nanoplasmon coupled optical resonance excitation (CORE) mechanism which permits enhanced surface plasmon on the substrate being evanescently coupled to Whispering Gallery mode optical resonance inside the spheroidal cell membrane microcavity. Theoretical model of the hypothesis is explained using coupled mode theory. In addition, preliminary result has been provided for the observation of early stage of transfection in a cancer cell.

We present the application of ellipsometry to the phase measurement of surface plasmon resonance (SPR) in biomolecular detection. In this work, the experimental setup for the SPR sensor was based on a custom-built rotating analyzer ellipsometer, which was equipped with a SPR cell and a microfluidic system. We investigate the sensitivity of SPR sensor which is dependent on the thickness and roughness of metal film, alignment of optical system, and stability of microfluidics. In the drug discovery process, to directly monitor the interaction of small molecule-protein, it is necessary to design a high-sensitivity SPR sensor with a sensitivity of greater than 1 pg/mm². Our sensor demonstrates a much better sensitivity in comparison to other SPR sensors based on reflectometry or phase measurements. The results of calibration indicate that the phase change, δ▵, had an almost linear response to the concentration of ethanol in the double-distilled water solutions. A quantitative analysis of refractive index variation was possible using the results of the ellipsometric model fits for the multilayered thin film on the gold film. Thus, this method is applicable not only to sensor applications, such as affinity biosensors, but also to highly sensitive kinetics for drug discovery. In this paper, we demonstrate how a custom-built rotating analyzer ellipsometer in the SPR condition can be used to directly obtain the interactions and binding kinetics of analytes (biotins, peptides) with immobilized ligand (streptavidin, antibody). We achieved a detection limit of lower than 1.0 x10^-7 RIU, which is the equivalent of 0.1 pg/mm².

Determination of whether nanoparticles accumulate in target or non-target tissues is critical in assessing a nanoparticle formulation for nanomedical purposes. There is an overwhelming need for a sensitive and efficient imaging-based method that can (1) detect small numbers of (even single) nanoparticles, (2) associate nanoparticle uptake with cell type, and (3) allow for rapid detection in large tissue samples. We propose a novel method for nanoparticle detection that utilizes an oligonucleotide "nanobarcode" conjugated to the nanoparticle surface, which amplifies the optical signal from a single nanoparticle via in situ PCR. Herein, we describe the design process of the nanobarcoding method.

We demonstrate surface plasmon-induced enhancements in optical imaging and spectroscopy on silver coated silicon nanocones which we call black silver substrate. The black silver substrate with dense and homogeneous nanocone forest structure is fabricated on wafer level with a mass producible nanomanufacturing method. The black silver substrate is able to efficiently trap and convert incident photons into localized plasmons in a broad wavelength range, which permits the enhancement in optical absorption from UV to NIR range by 12 times, the visible fluorescence enhancement of ~30 times and the NIR Raman scattering enhancement factor up to ~10⁸. We show a considerable potential of the black silver substrate in high sensitivity and broadband optical sensing and imaging of chemical and biological molecules.one)

A newly emerging field in bioanalytics based on biomolecular binding detected label-free at single metal nanoparticles is introduced. Thereby particles which show the effect of localized surface plasmon resonance (LSPR) are used as plasmonic transducers. They change their spectroscopic properties (a band in the UV-VIS range) upon binding of molecules. This effect is even observable at the single nanoparticle level using micro spectroscopy and presents the base for a new field of single particle bioanalytics with the promise of highly parallel and miniaturized sensor arrays. The paper describes this approach and shows first result from our work regarding the detection of DNA binding at single nanoparticle sensors.

Metallic nanoparticles have drawn much interest due to their distinct plasmonic characteristics especially in imaging and sensing applications. Surface plasmon resonance (SPR) based biosensors have evolved in many ways, among which sensitivity enhancement towards molecular sensing capability came up with strategies to overcome the hard limit of the intrinsic sensitivity of gold thin film. Recently adoption of signal contrast materials has proven successful in biochemical sensing applications. This study employs gold-SiO2 core-shell nanoparticles (CSNPs) as a strong SPR signal contrast agents. To reveal the underlying physics for the contrast mechanism, the particle characteristics were analytically evaluated in terms of light interaction coefficients. We experimentally demonstrate the effect of the CSNPs by applying them to acquire enhanced signal in DNA hybridization sensing scheme. We also applied gold nanowire grating structure on conventional gold thin film to further amplify the intrinsic sensitivity, where localized surface plasmon and locally amplified evanescent fields take parts. The results suggest that CSNPs and the grating structure cooperatively enhance the sensitivity and the role of nanowire gratings was analyzed with numerical methods to allow optimum sensitivity enhancement in terms of fill factor variations. The effects of field localization, amplification and enlarged signature of CSNPs are also discussed.

Bacteria, viruses and parasites elimination from human environment is one of the most important problem, extensively studied by many groups. The growing resistance to commonly used disinfection and/or sterilization methods and antibiotics, is one of the major problem in the health care sector. Nanomaterials with tailored antimicrobial features may find applications in this field. One of the promising application of nanomaterials is the possibility to enhance the antimicrobial photodynamic therapy (APDT), which combines a nontoxic photoactive dye - photosensitizer and nanomaterials properties. This paper focused on the examination of optical and antibacterial properties of silica- and titania-based nanopowders doped with silver and photosensitizer - Photolon. Various concentration of Photolon and nanomaterials have been prepared in order to examine the fluorescence enhancement and resulting better antibacterial activity. It was proved that the fluorescence intensity of Photolon increased, depending on silver concentration. Antibacterial study showed that silver doped silica and titania nanoparticles revealed antibacterial activity, but in the presence of Photolon, the antibacterial activity of materials is more effective.

The purpose of this paper is to analyze the current evolutions on nanotechnology and its applications on cancer theragnostics.Rapid advances and emerging technologies in nanotechnology are having a profound impact on cancer treatment. Applications of nanotechnology, which include liposomes, nanoparticles, polymeric micelles, dendrimers, nanocantilever, carbon nanotubes and quantum dots have significantly revolutionized cancer theragnostics. From a pharmaceutical viewpoint, it is critical that the biodistribution of active agents has to be controlled as much as possible. This aspect is vital in order to assure the proper efficiency and safety of the anticancer agents. These biocompatible nanocomposites provide specific biochemical interactions with receptors expressed on the surface of cancer cells. With passive or active targeting strategies, an increased intracellular concentration of drugs can be achieved in cancer cells , while normal cells are being protected from the drug simultaneously. Thus, nanotechnology restricts the extent of the adverse effects of the anticancer therapy. Treatment for metastatic breast cancer, sarcoma in AIDS patients, ovarian and lung cancer is already on market or under final phases of many clinical trials, showing remarkable results. As nanotechnology is perfected, side effects due to normal cell damage will decrease, leading to better results and lengthening patient's survival.

Endocrine disrupting compounds (EDCs) such as bisphenol A (BPA) and female hormone Estrone are especially prevalent in surface and waste-waters in nano-molar concentrations and therefore, there is a need for sensitive analytical device for their monitoring. We have designed a miniature, low cost and fast surface plasmon resonance (SPR) imaging liquid sensor based on the angular interrogation using Kretschmann configuration with diverged incident monochromatic light. During this paper we present a surface plasmon resonance imaging (SPRI) biosensor to detect EDCs such as BPA and estrone. A pattern of SPR line which is dark intensity line on bright area was reflected at angles range depending on the dielectric constant of the analye: Rabbit Anti-Estrone polyclonal IgG + Estrone 11-MUA attached to the silver or non-specific sensing of BPA in water with nanoprecision. For analyzing the SPR signals we used an efficient detection algorithm based on Radon Transform with less sensitivity to laser speckle noise and nonuniformity of the illumination.