This PDF file contains the front matter associated with SPIE Proceedings Volume 9721 including the Title Page, Copyright information, Table of Contents, Introduction, and Conference Committee listing.

Light can be made to do the work. Imagine tweezers made out of light. Such optical tweezers can trap and move materials noninvasively at length scales ranging from tens of nanometers to tens of micrometers, and so have provided unprecedented access to physical, chemical and biological processes on a microscale. Since a light beam can carry angular momentum it is possible to use optical tweezers to exert torques to twist or rotate nano and microscopic objects. These optical rotors can be used to map the mechanical properties of cells. They can also be used in biotechnology and optomechanics.

Employing vibrational spectroscopy (IR-absorption and Raman spectroscopy) allows for the labelfree detection of molecular specific fingerprints of inorganic, organic and biological substances. The sensitivity of vibrational spectroscopy can be improved by several orders of magnitude via the application of plasmonic active surfaces. Within this contribution we will discuss two such approaches, namely surface enhanced Raman spectroscopy (SERS) as well as surface enhanced IR absorption (SEIRA). It will be shown that SERS using metal colloids as SERS active substrate in combination with a microfluidic lab-on-a-chip (LOC) device enables high throughput and reproducible measurements with highest sensitivity and specificity. The application of such a LOC-SERS approach for therapeutic drug monitoring (e.g. quantitative detection of antibiotics in a urine matrix) will be presented. Furthermore, we will introduce innovative bottom-up strategies to prepare SERS-active nanostructures coated with a lipophilic sensor layer as one-time use SERS substrates for specific food analysis (e.g. quantitative detection of toxic food colorants). The second part of this contribution presents a slit array metamaterial perfect absorber for IR sensing applications consisting of a dielectric layer sandwiched between two metallic layers of which the upper layer is perforated with a periodic array of slits. Light-matter interaction is greatly amplified in the slits, where also the analyte is concentrated, as the surface of the substrate is covered by a thin silica layer. Thus, already small concentrations of analytes down to a monolayer can be detected by refractive index sensing and identified by their spectral fingerprints with a standard mid-infrared lab spectrometer.

Localized surface Plasmon Resonance (LSPR) is a nanoscale phenomenon which presents strong resonance associated with noble metal nanostructures. This plasmon resonance based technology enables highly sensitive detection for chemical and biological applications. Recently, we have developed a plasmon field effect transistor (FET) that enables direct plasmonic-to-electric signal conversion with signal amplification. The plasmon FET consists of back-gated field effect transistor incorporated with gold nanoparticles on top of the FET channel. The gold nanostructures are physically separated from transistor electrodes and can be functionalized for a specific biological application. In this presentation, we report a successful demonstration of a model system to detect Con A proteins using Carbohydrate linkers as a capture molecule. The plasmon FET detected a very low concentration of Con A (0.006 mg/L) while it offers a wide dynamic range of 0.006-50 mg/L. In this demonstration, we used two-color light sources instead of a bulky spectrometer to achieve high sensitivity and wide dynamic range. The details of two-color based differential measurement method will be discussed. This novel protein-based sensor has several advantages such as extremely small size for point-of-care system, multiplexing capability, no need of complex optical geometry.

We report a novel nanophotonic biosensor surface capable of both colorimetric detection and Raman-scattered detection of DNA infection markers at extreme sensitivities. Combining direct-write lithography, dip-pen nanolithography based DNA patterning, and molecular self-assembly, we create molecularly-active plasmonic nanostructures onto which metallic nanoparticles are located via DNA-hybridization. Arraying these structures enables optical surfaces that change state when contacted by specific DNA sequences; shifting the surface color while simultaneously generating strong Raman-scattering signals. Patterning the DNA markers onto the plasmonic surface as micro-scale symbols results in easily identifiable color shifts, making this technique applicable to multiplexed lab-on-a-chip and point-of-care diagnostic applications.

Herein we describe promising results from the combination of fluorescent lifetime imaging microscopy (FLIM) and diffusion reflection (DR) medical imaging techniques. Three different geometries of gold nanoparticles (GNPs) were prepared: spheres of 20nm diameter, rods (GNRs) of aspect ratio (AR) 2.5, and GNRs of AR 3.3. Each GNP geometry was then conjugated using PEG linkers estimated to be 10nm in length to each of 3 different fluorescent dyes: Fluorescein, Rhodamine B, and Sulforhodamine B. DR provided deep-volume measurements (up to 1cm) from within solid, tissue-imitating phantoms, indicating GNR presence corresponding to the light used by recording light scattered from the GNPs with increasing distance to a photodetector. FLIM imaged solutions as well as phantom surfaces, recording both the fluorescence lifetimes as well as the fluorescence intensities. Fluorescence quenching was observed for Fluorescein, while metal-enhanced fluorescence (MEF) was observed in Rhodamine B and Sulforhodamine B – the dyes with an absorption peak at a slightly longer wavelength than the GNP plasmon resonance peak. Our system is highly sensitive due to the increased intensity provided by MEF, and also because of the inherent sensitivity of both FLIM and DR. Together, these two modalities and MEF can provide a lot of meaningful information for molecular and functional imaging of biological samples.

In this presentation, we explore the feasibility of plasmonic nanohole-based sub-diffraction-limited nanoscopy for biomolecular imaging. The technique utilizes near-field distribution localized by surface plasmon localization on metallic nanoholes which is used to sample molecular fluorescence. The optimum geometry of nanohole arrays was determined by numerical analysis. The localization sampling was applied to reconstructing sub-diffraction-limited images of gliding microtubules with a 76 nm effective resolution in the lateral direction. Extraordinary light transmission was also employed to address enhancement of axial resolution using nanohole arrays, based on which extraction of gliding motions of bacteria was demonstrated with an axial resolution down to 50 nm.

Localized surface plasmon resonance has been extensively investigated for biochemical sensor applications. In traditional localized surface plasmon resonance biosensors, resonance spectra were measured in the reflection or transmission from the nanostructure devices. In this work, we demonstrate a new surface plasmon resonance sensor platform with which the localized surface plasmon resonance and shift were measured by using a CCD imager instead of using an optical spectrometer. In additional to the metal nanostructures which support localized plasmon resonance, we pattern the nanostructures into diffraction gratings with super-wavelength grating periods. The nanostructure diffraction gratings support localized plasmon resonance and also diffract localized plasmon resonance radiations into non-zeroth order diffractions. Plasmon resonance spectrum and shift are measured with a CCD imager in one of the diffraction orders. The new plasmon resonance spectrometer sensor combines the functions of sensing and spectral analysis into one apparatus and is capable of real-time visualization of the biochemical bonding process with an imager.

We designed and fabricated microscale lens arrays on a flexible substrate. The flexibility of the substrate allows for wide field of view imaging as well as optical focus scanning. Fresnel zone plates (FZPs), which are compact and lightweight, are used as microlenses for focusing. The arrangement of FZPs on flexible substrate can be reconfigured to maximize FOV. Tunable focus can also be achieved by stretching the FZPs laterally. In addition, the lightweight microlenses can be actuated to scan the focus axially. The lenses have a wide range of applications including displays, contact lenses, microscopy, surveillance and optical communications. The diameter of the microlenses ranges from 100 to 500 µm. The thickness of the lenses is 100 µm. Unlike refractive and reflective lenses, the focusing capability of FZPs is achieved via diffraction. FZPs consist of alternating black and white zones to modulate the phase of the incident light. The light diffracted from edge of the regions to achieve multiple focus. Most of the energy is diffracted into the first focus. The dark regions are made of silicon nanowires which are highly absorbent for visible spectrum. Standard processes, including wet and dry etching, are used to etch silicon substrate and form nanowires. The white zones are designed for both reflective and transmissive lenses. The lenses are implemented on PDMS as flexible substrate. The silicon nanowires are embedded into PDMS so that the shape of individual lens as well as the arrangement of the array can be reconfigured. In this article, we report our design, fabrication process and experiments.

We present surface plasmon enhanced fluorescence microscopy with random spatial sampling using patterned block of silver nanoislands. Rigorous coupled wave analysis was performed to confirm near-field localization on nanoislands. Random nanoislands were fabricated in silver by temperature annealing. By analyzing random near-field distribution, average size of localized fields was found to be on the order of 135 nm. Randomly localized near-fields were used to spatially sample F-actin of J774 cells (mouse macrophage cell-line). Image deconvolution algorithm based on linear imaging theory was established for stochastic estimation of fluorescent molecular distribution. The alignment between near-field distribution and raw image was performed by the patterned block. The achieved resolution is dependent upon factors including the size of localized fields and estimated to be 100-150 nm.

Cortisol, a biomarker of stress, has recently been shown to have potential in evaluating the physiological state of individuals diagnosed with stress-related conditions including chronic fatigue syndrome. Noninvasive techniques to extract biomarkers from the body are a topic of considerable interest. One such technique to achieve this is known as reverse iontophoresis (RI) which is capable of extracting biomolecules through the skin. Unfortunately, however, the extracted levels are often considerably lower in concentration than those found in blood, thereby requiring a very sensitive analytical method with a low limit of detection. A promising sensing approach, which is well suited to handle such samples, is Surface Plasmon Resonance (SPR) spectroscopy. When coupled with aptamer modified surfaces, such sensors can achieve both selectivity and the required sensitivity. In this study, fabrication and characterization of a RIbased SPR biosensor for the measurement of cortisol has been developed. The optical mount and diffusion cell were both fabricated through the use of 3D printing techniques. The SPR sensor was configured to employ a prism couplerbased arrangement with a laser generation module and CCD line sensor. Cortisol-specific DNA aptamers were immobilized onto a gold surface to achieve the necessary selectivity. For demonstration purposes, cortisol was extracted by the RI system using a skin phantom flow system capable of generating time dependent concentration profiles. The captured sample was then transported using a micro-fluidic platform from the RI collection site to the SPR sensor for real-time monitoring. Analysis and system control was accomplished within a developed LabVIEW® program.

Understanding the cellular signaling and function at the nano-bio interface can pave the way towards developing next-generation smart diagnostic tools. From this perspective, limited reports detail so far the cellular and subcellular forces exerted by bacterial cells during the interaction with abiotic materials. Nanowire arrays with high aspect ratio have been used to detect such small forces. In this regard, live force measurements were performed ex-vivo during the interaction of Xylella fastidiosa bacterial cells with InP nanowire arrays. The influence of nanowire array topography and surface chemistry on the response and motion of bacterial cells was studied in detail. The nanowire arrays were also functionalized with different cell adhesive promoters, such as amines and XadA1, an afimbrial protein of X.fastidiosa. By employing the well-defined InP nanowire arrays platform, and single cell confocal imaging system, we were able to trace the bacterial growth pattern, and show that their initial attachment locations are strongly influenced by the surface chemistry and nanoscale surface topography. In addition, we measure the cellular forces down to few nanonewton range using these nanowire arrays. In case of nanowire functionalized with XadA1, the force exerted by vertically and horizontally attached single bacteria on the nanowire is in average 14% and 26% higher than for the pristine array, respectively. These results provide an excellent basis for live-cell force measurements as well as unravel the range of forces involved during the early stages of bacterial adhesion and biofilm formation.

Nanoparticle suspensions are used in numerous biomedical applications ranging from sensing and diagnostics to in vivo therapeutic agents and drug delivery mechanisms. One key challenge in developing these technologies is engineering particles that remain stable in the presence of physiological salt concentrations and different pH regimes encountered in applications. Here, we show an approach for high-throughput characterization of nanoparticle stability by directly measuring the interaction energy profiles between nanoparticles and surfaces. As nanoparticles are trapped and propelled along an optical waveguide, they scatter light. Our technique takes advantage of the confined Brownian motion exhibited by the particles as they fluctuate about the equilibrium position between the optical and particle-surface interaction forces. In this way, unlike colloidal probe atomic force microscopy, this technique is capable of making measurements that are not limited by thermal noise, and capable of mapping interaction energy profiles on the sub-kT scale, driven by sub-pN forces. We demonstrate direct measurement of the interactions between protein-coated gold nanoparticles with 50 nm diameters and surfaces in a variety of experimental conditions including changes in specific ions present, overall ionic strength and pH, giving insight into the dynamics of these biologically relevant systems at the nanoscale. These direct measurements on particles with sub-100 nm diameters offer new insights into suspension stability missed by indirect measurements such as absorbance spectroscopy, zeta-potential, and dynamic light scattering, and allow for the detailed study of sub-populations in a heterogeneous sample. Additionally, the sub-pN force resolution makes this a suitable platform for fundamental biophysical studies.

In our study we aim to develop a new, simple and non-invasive method to detect and to treat atherosclerosis. We use gold nanoparticles (GNPs) combined with the diffusion reflection (DR) method to demonstrate the detection of vulnerable atherosclerotic plaques. Our method is based on the fact that macrophages are a major component in the vulnerable plaque and are able to uptake metal nanoparticles that can be discovered by the DR system. Moreover, it is well known that high density lipoprotein (HDL) reduces ASVD by inhibiting pro-inflammatory factors, enabling the specific treatment of atherosclerosis.

The cross-coupling reactions have been used in C-C bond formation which can be used extensively in optoelectronic materials for organic light emitting diode (OLED), organic photovoltaics and chemical biosensing. Here, we report twofold geminal C-C bond formation at 1,1-dibromoolefins via cross-coupling reactions of aromatic boronic esters over Pd catalysts for multiple topological configurations of π-conjugated molecules. We employ a series of recipes from a precursor toolbox to produce π-conjugated macrocycles, conjugated dendrimers, 1-dimensional linear conjugated polymers, 2-dimensional conjugated microporous polymers (CMPs) and crosslinking conjugated polymer nanoparticles (CCPNs). The π-conjugated macrocycles, dendrimers and 1-D polymers show characteristic aggregation-induced emission properties. 2-D conjugated microporous polymers possess unique porosity of 2-3 nm. This universal strategy toward definite topological configurations of π-conjugated molecules enables efficient coupling of aryl bromides with various coupling partners under mild conditions affording multiple topological conjugated systems with abundant optical and optoelectronic interest.

We present optoelectronic investigation of in vitro interactions of whole human blood with different nanodiamond biomarkers. Plasmo-chemical modifications of detonation nanodiamond particles gives the possibility for controlling their surface for biological applications. Optical investigations reveal the biological activity of nanodiamonds in blood dependent on its surface termination. We compare different types of nanodiamonds: commercial non-modified detonation nanodiamonds, and nanodiamonds modified by MW PACVD method with H₂-termination, and chemically modified nanodiamond with O₂-termination. The absorption spectra, and optical microscope investigations were conducted. The results indicate haemocompatibility of non-modified detonation nanodiamond as well as modified nanodiamonds, which enables their application for drug delivery, as well as sensing applications.

Background/Aim: In atherosclerosis stable and vulnerable atherosclerotic plaque types are distinguished that behave differently concerning rupture, thrombosis and clinical events. The stable are rich in M2 macrophages. The unstable are rich in inflammatory M1 macrophages and are highly susceptible to rupture, setting patients at risk for thrombotic events when they undergo invasive diagnosis such as coronary angiography. Therefore, novel approaches for non-invasive detection and classification of vulnerable plaques in vivo are needed. Whereas classical approaches fail to differentiate between both plaque types, a new biophotonic method (combination of the diffusion reflection (DR) method with flow cytometry (FCM) or image cytometry (IC)) to analyze gold nanoparticle (GNP) loading of plaques could overcome this limitation. Methods: Two types of GNP were used three variants of gold nanorods (GNRI with 40x18 nm, II 65x25 nm and III 52x13 nm in size) and gold nanospheres (GNS with an average diameter of 18.5 nm). The GNS had an absorption peak at 520 nm and the GNR at 630 nm. Monocytes were isolated from human buffy blood samples, differentiated into macrophages and their subtypes and labelled with GNR and GNS for 3 and 24 h. GNS and GNR loading were determined by FCM and/or IC. Macrophages within tissue-like phantoms were analyzed by the DR system. Results: After GNR labelling of macrophages the FCM light scatter values increased up to 3.7 fold and the DR slope changed from an average slope of 0.196 (macrophages only) to an average slope of 0.827 (macrophages labelled with GNR). But, GNRIII did not present much higher DR slopes than the control phantoms, indicating that macrophages take up GNRIII in a lower amount than GNRI or II. IC and microscopy showed that all particle variants were taken up by the cells in a heterogeneous fashion. Conclusion and outlook: The combination of FCM and DR measurements provides a potential novel, highly sensitive and non-invasive method for the identification of atherosclerotic vulnerable plaques, aimed to develop a potential tool for in vivo tracking. Further experiments will show, if different macrophage subtypes (M1 or M2) take up the particles differently and may thereby serve to distinguish stable from vulnerable plaques.

In this paper we present gold nanoparticles coated with silicon that switch the order between the scattering and the absorption magnitude at the resonance peak and tune the plasmon resonance over the spectrum. This is obtained by modifying the refractive index of the silicon coating of the nanoparticle by illuminating it with a pumping light due to the plasma dispersion effect in silicon. We also report how changing the diffraction limited point spread function through the utilization of plasma dispersion effect of the above mentioned silicon coated nanoparticles allows doing imaging with sub wavelength resolution. The plasma dispersion effect can increase the absorption coefficient of the silicon, when illuminated with a focused laser beam and as explained above it can also tune the absorption versus scattering properties of the nanoparticle. Due to the Gaussian nature of the laser illumination which has higher intensity at its peak, the plasma dispersion effect is more significant at the center of the illumination. As a consequence, the reflected light from probe beam at the near infra-red region has a sub wavelength dip that overlaps with the location of the pump illumination peak. This dip has a higher spatial frequency than an ordinary Gaussian, which enables to achieve super resolution.

In recent years, optical super-resolution by microspheres and microfibers emerged as a new paradigm in nanoscale label-free and fluorescence imaging. However, the mechanisms of such imaging are still not completely understood and the resolution values are debated. In this work, the fundamental limits of super-resolution imaging by high-index barium-titanate microspheres and silica microfibers are studied using nanoplasmonic arrays made from Au and Al. A rigorous resolution analysis is developed based on the object’s convolution with the point-spread function that has width well below the conventional (~λ/2) diffraction limit, where λ is the illumination wavelength. A resolution of ~λ/6-λ/7 is demonstrated for imaging nanoplasmonic arrays by microspheres. Similar resolution was demonstrated for microfibers in the direction perpendicular to the fiber axis with hundreds of times larger field-of-view in comparison to microspheres. Using numerical solution of Maxwell’s equations, it is shown that extraordinary close point objects can be resolved in the far field, if they oscillate out of phase. Possible super-resolution using resonant excitation of whispering gallery modes is also studied.

Energy-time entangled photon pairs exhibit at the same time narrowband and short time features. We show how to make use of those quantum properties to realize measurements beyond the capabilities of classical devices. As proof of principle experiment we show imaging through a scattering medium by selecting the ballistic photons only, using optical coincidence measurements.

The intrinsic near-infrared photoluminescence (fluorescence) of single-walled carbon nanotubes exhibits unique photostability, narrow bandwidth, penetration through biological media, environmental sensitivity, and both chromatic variety and range. Biomedical applications exploiting this large family of fluorophores will require the spectral and spatial resolution of individual (n,m) nanotube species’ fluorescence and its modulation within live cells and tissues, which is not possible with current microscopy methods. We present a wide-field hyperspectral approach to spatially delineate and spectroscopically measure single nanotube fluorescence in living systems. This approach resolved up to 17 distinct (n,m) species (chiralities) with single nanotube spatial resolution in live mammalian cells, murine tissues ex vivo, and zebrafish endothelium in vivo. We anticipate that this approach will facilitate multiplexed nanotube imaging in biomedical applications while enabling deep-tissue optical penetration, exceptional photostability, and single-molecule resolution in vivo.

Surface plasmon resonant (SPR) phenomenon is widely researched for various purposes, among which biomedical sensing is getting more attentions as they are suitable for surface functionalization acting as a bio recognition element to detect different biological infections. The common method of surface resonant is propagating SPR such as reflection method. Another method which is widely used for SPR is localized SPR which use nanostructures in thin metal. Various structures such as slit only, slit- groove and slit-multiple groove are used for generation of SPR and obtaining the optimum optical transmittance through the structure. The number and position of slits and grooves affect transmittance through the structure. In this paper we propose a new structure of cross slit-grooves structure, which includes slit-groove structure in grid form. The slit-grooves structures are arranged in such a way that it forms symmetrical structure in two dimension with slit and groove and hence the transmittance with cross slit-grooves structure increases significantly. The cross slit-grooves structure takes the advantage of symmetrical slit and groove by using both dimensional structures for generating SPR which increases the transmittance through the structure. A comparison of proposed slit-grooves grid structure with straight slit-grooves structure is carried out to show the increase in transmittance through the cross slit-grooves grid structure. Plane wavelength of 400 nm to 900 nm is used for the analysis of transmittance through the Ag slit-grooves grid structures with glass substrate. We also measure the change in transmittance with change in refractive index, which can be helpful for measuring different chemical analytes, and hence can be used for different chemical and biosensors applications.

Various techniques for recovering optical parameters were developed over the years. However each has its limitations, constraints and disadvantages (e.g. accuracy, computational speed, sample assembly, distinguishing between the different parameters, etc.). This research suggests an optical technique for extracting the reduced scattering coefficient (μs') of substances by examining the light transmission through or reflection from them. It uses the multiple planes Gerchberg- Saxton (G-S) algorithm to reconstruct the light phase created by the substance. At the end of the algorithm, μs' can be estimated from the standard deviation (STD) of the retrieved phase of the reemitted light. We will use the theory to compute the phase’s STD that directly correlated to the optical properties of different substances. Two possible applications for this technique, out of many others, are nanoparticles (NPs) penetration depth determination, for promoting topical medications, and detection of milk components quantitative signature as en route to milk content monitoring tool. For the former application, three materials were fabricated into NPs and all presented an activity enhancement with their size reduction. Then the NPs were applied on tissues and detected by our technique. For the latter, different milk content concentrations were examined resulting with different STD values suggesting it can be used as indicator for the milk component concentrations.

Functional nanoscale materials are being extensively investigated for applications in biology and medicine and are ready to make significant contributions in the realization of exciting advancements in diverse areas of diagnostics and therapeutics. Aiming for more accurate, efficient, non-invasive and fast diagnostic tools, the use of near-infrared (NIR) light in the range of the 1st and 2nd biological window (NIR-I: 0.70-0.95 µm; NIR-II: 1.00-1.35 µm) provides deeper penetration depth into biological tissue, better image contrast, reduced phototoxicity and photobleaching. Consequently, NIR-based bioimaging became a quickly emerging field and manifold new NIR-emitting bioprobes have been reported. Since commercially available microscopes are not optimized for this kind of NPs, a new microscopy hyperspectral confocal imager has been developed to cover a broad spectral range (400 to 1700 nm) with high spectral resolution. The smallest spectral variation can be easily monitored thanks to the high spectral resolution (as low as 0.2 nm). This is possible thanks to a combination of an EMCCD and an InGaAs camera with a high resolution spectrometer. An extended number of NPs can be excited with a Ti:Sapphire laser, which provides tunable illumination within 690-1040 nm. Cells and tissues can be mapped in less than 100 ms, allowing in-vivo imaging. As a proof of concept, here we present the preliminary results of the spatial distribution of the fluorescence signal intensity from lanthanide doped nanoparticles incorporated into a system of biological interest. The temperature sub-mm gradient – analyzing the spectral features so gathered through an all-optical route is also thoroughly discussed.

Non-radiative Excitation Fluorescence Microscopy (NEFM) constitutes a new way to observe biological samples beyond the diffraction limit. Non-radiative excitation of the samples is achieved by coating the substrate with donor species, such as quantum dots (QDs). Thus the dyes are not excited directly by the laser source, as in common fluorescence microscopy, but through a non-radiative energy transfer. To prevent dewetting of the donor film, we have recently implemented a silanization process to covalently bond the QDs on the substrate. An homogeneous monolayer of QDs was then deposited on only one side of the coverslips. Atomic force microscopy was then used to characterize the QD layer. We highlight the potential of our method through the study of Giant Unilamellar Vesicles (GUVs) labeled with DiD as acceptor, in interaction with surface functionalized with poly-L-lysine. In the presence of GUVs, we observed a quenching of QDs emission, together with an emission of DiD located in the membrane, which clearly indicated that non-radiative energy transfer from QDs to DiD occurs.

In this paper we aim to experimentally verify a speckle based technique for non-contact measurement of glucose concentration in blood stream while the vision for the final device aims to contain a single wristwatch-style device containing an AC (alternating) electro-magnet generated by a solenoid, a laser and a camera. The experiments presented in work are performed in-vitro in order to verify the effects that are responsible for the operation principle. When a glucose substance is inserted into a solenoid generating an alternating magnetic field it exhibits Faraday rotation which affects the temporal changes of the secondary speckle patterns distribution. The temporal frequency resulting from the AC magnetic field was found to have a lock-in amplification role which increased the observability of the relatively small magneto-optic effect. Experimental results to support the proposed concept are presented.

Brain tumors are the second leading cause of cancer-related deaths in children, after leukemia. Patients with cancer in the central nervous system have a very low recovery rate. Today known imaging and cytology techniques are not always sensitive enough for an early detection of both tumor and its metastatic spread, moreover the detection is generally limited, reviewer dependent and takes a relatively long time. Medulloblastoma (MB) is the most common malignant brain tumor in children. The aim of our talk is to present the frequency domain fluorescence lifetime imaging microscopy system as a possible method for an early detection of MB and its metastatic spread in the cerebrospinal fluids within the pediatric population.

Optical wide-field imaging of sub-diffraction limit nanostructures is of interest in a wide array of applications. In applications where the nanostructures to be visualized are well isolated, a high enough optical contrast is sufficient to detect these. Here we demonstrate a technique to visualize nanoscale features, such as grain boundaries in Chemical Vapor Deposited (CVD) single layer graphene, which are just a few atom length defects, using regular bright field optical microscopy. This remarkably low lateral length scale was imaged using of a special thin film structure consisting of a water-soluble thin film layer deposited on a metal substrate, which produces a strong color change as a function of the film thickness. Small local water transport differences in the graphene layer result in thickness variation of the underlying thin film due to its solubility in water and produces color contrast readily observable under a normal brightfield optical microscope with the naked eye. We demonstrate the use of this technique for direct optical visualization of grain boundaries in graphene as wide as 2-5 nm and sub-100 nm photoresist lines. By using super-resolution image processing algorithms, we may be able to detect structure even smaller in size than currently achieved. We believe that this technique can be extended to single molecule detection.

This work presents the use of flickering nanoparticles for imaging biological samples. The method has high noise immunity, and it enables the detection of overlapping types of GNPs, at significantly sub-diffraction distances, making it attractive for super resolving localization microscopy techniques. The method utilizes a lock-in technique at which the imaging of the sample is done using a time-modulated laser beam that match the number of the types of gold nanoparticles (GNPs) that label a given sample, and resulting in the excitation of the temporal flickering of the scattered light at known temporal frequencies. The final image where the GNPs are spatially separated is obtained using post processing where the proper spectral components corresponding to the different modulation frequencies are extracted. This allows the simultaneous super resolved imaging of multiple types of GNPs that label targets of interest within biological samples. Additionally applying the post-processing algorithm of the K-factor image decomposition algorithm can further improve the performance of the proposed approach.

Skin cancer detection at its early stages has been the focus of a large number of experimental and theoretical studies during the past decades. Among these studies two prominent approaches presenting high potential are reflectometric sensing at the THz wavelengths region and polarimetric imaging techniques in the visible wavelengths. While THz radiation contrast agent and source of sensitivity to cancer related tissue alterations was considered to be mainly the elevated water content in the cancerous tissue, the polarimetric approach has been verified to enable cancerous tissue differentiation based on cancer induced structural alterations to the tissue. Combining THz with the polarimetric approach, which is considered in this study, is examined in order to enable higher detection sensitivity than previously pure reflectometric THz measurements. For this, a comprehensive MC simulation of radiative transfer in a complex skin tissue model fitted for the THz domain that considers the skin`s stratified structure, tissue material optical dispersion modeling, surface roughness, scatterers, and substructure organelles has been developed. Additionally, a narrow beam Mueller matrix differential analysis technique is suggested for assessing skin cancer induced changes in the polarimetric image, enabling the tissue model and MC simulation to be utilized for determining the imaging parameters resulting in maximal detection sensitivity.

Super-resolution localization microscopy methods rely on accurate and fast localization algorithms. We introduce a simple algorithm for the localization of imaged objects based on the search of the best-correlated center. This approach yields tracking accuracies that are comparable to those of Gaussian fittings in typical low signal-to-noise ratios, but with 6× faster execution. The algorithm can be adapted to localize objects that do not exhibit radial symmetry or have to be localized in higher dimensional spaces.

Human tissue is one of the most complex optical media since it is turbid and nonhomogeneous. We suggest a new optical method for sensing physiological tissue state, based on the collection of the ejected light at all exit angles, to receive the full scattering profile. We built a unique set-up for noninvasive encircled measurement. We use a laser, a photodetector and tissues-like phantoms presenting different diameters and different reduced scattering coefficients. Our method reveals an isobaric point, which is independent of the optical properties and linearly depends on the exact tissue geometry. Furthermore, we present the angular distribution of cylindrical silicon based phantoms containing blood vessels in different diameters, in order to sense physiological tissue state. We show, for the first time, by simulation and experiments, that the vessel diameter influences on the full scattering profile. In addition, we found higher reflection intensity for larger vessel diameters, in accordance to the shielding effect. These findings can be useful for biomedical applications such as non-invasive and simple diagnostic of the fingertip joint, ear lobe and pinched tissues.