Molecular Structure and Interactions of Lipids in the Outer Membrane of Living Cells Based on Surface-Enhanced Raman Scattering and Liposome Models.

The distribution and interaction of lipids determine the structure and function of the cellular membrane. Surface-enhanced Raman scattering (SERS) is used for selective molecular probing of the cell membrane of living fibroblast cells grown adherently on gold nanoisland substrates across their whole contact areas with the substrate, enabling mapping of the membrane's composition and interaction. From the SERS data, the localization and distribution of different lipids and their interactions, together with proteins in the outer cell membrane, are inferred. Interpretation of the spectra is mainly supported by comparison with the spectra of model liposomes composed of phosphatidylcholine, sphingomyelin, and cholesterol obtained on the same gold substrate. The interaction of the liposomes with the substrate differs from that with gold nanoparticles. The SERS maps indicate colocalization of ordered lipid domains with cholesterol in the living cells. They support the observation of ordered membrane regions of micrometer dimensions in the outer leaflet of the cell membrane that are rich in sphingomyelin. Moreover, the spectra of the living cells contain bands from the groups of the lipid heads, phosphate, choline, and ethanolamine, combined with those from membrane proteins, as indicated by signals assigned to prenyl attachment. Elucidating the composition and structure of lipid membranes in living cells can find application in many fields of research.

Polarization state generation and measurement with a single metasurface.

The constituent elements of metasurfaces may be designed with explicit polarization dependence, making metasurfaces a fascinating platform for new polarization optics. In this work we show that a metasurface grating can be designed to produce arbitrarily specified polarization states on a set of defined diffraction orders given that the polarization of the incident beam is known. We also demonstrate that, when used in a reverse configuration, the same grating may be used as a parallel snapshot polarimeter, requiring a minimum of bulk polarization optics. We demonstrate its use in measuring partially polarized light, and show that it performs favorably in comparison to a commercial polarimeter. This work is of consequence in any application requiring lightweight, compact, and low-cost polarization optics, polarimetry, or polarization imaging.

Efficient interfacing photonic and long-range dielectric-loaded plasmonic waveguides.

Long-range dielectric-loaded surface plasmon-polariton waveguides (LR-DLSPPWs) operating at telecom wavelengths are efficiently (end-fire) interfaced with photonic waveguides by taking advantage of very similar lateral mode field profiles in these waveguide configurations. The LR-DLSPPWs are formed by 1-µm-high and 1-µm-wide polymer ridges fabricated atop 15-nm-thin and 500-nm-wide gold stripes supported by a 289-nm-thick ormoclear polymer deposited on a low-index (1.34) layer of cytop, whereas gold stripes are absent in the photonic waveguide configuration that is identical to the plasmonic one in all other respects. The coupling efficiency between LR-DLSPPWs and photonic waveguides is numerically estimated to be 97%, decreasing by only a few percents for non-centered gold stripes (as long as a gold stripe is kept inside the polymer ridge). The fabricated LR-DLSPPWs coupled to photonic waveguides are first characterized using amplitude- and phase-resolved near-field imaging of propagating radiation that reveals very similar mode field distributions in these waveguides as well as their efficient interfacing. The coupling efficiency is then experimentally characterized using the cutback approach resulting in an average level of 75% per interface, while the LR-DLSPPW mode propagation length is estimated to be on average 0.3 mm. Possible reasons for differences between experimental and simulation results are discussed, indicating that a 3-nm-thin titanium layer (used for improving adhesion between gold and ormoclear) introduces substantial mode absorption. The results obtained open new perspectives for realization of hybrid photonic-plasmonic components and circuits.

Polarization-selective coupling to long-range surface plasmon polariton waveguides.

We demonstrate polarization-selective coupling from an optical fiber to long-range surface plasmon polariton waveguide modes using plasmonic antenna arrays. The arrays allow the sorting of two distinct (not necessarily orthogonal) polarizations to counter-propagating waveguide modes. The polarization-selective behavior of the devices is described by a compact formalism based on Stokes vectors that offers a clear graphical representation of the response. We experimentally observe polarization-controlled switching and unidirectional coupling with extinction ratios greater than 30 dB and coupling efficiencies comparable to those of a conventional grating coupler.

Data transmission in long-range dielectric-loaded surface plasmon polariton waveguides.

We demonstrate the data transmission of 10 Gbit/s on-off keying modulated 1550 nm signal through a long-range dielectric-loaded surface plasmon polariton waveguide structure with negligible signal degradation. In the experiment the bit error rate penalties do not exceed 0.6 dB over the 15 nm wavelength range and received optical power between -7 and 3 dBm.

Vapor-Phase Contribution to Laser-Induced Plasma Emission of Magnesium in Liquid Aluminum.

Liquid aluminum containing the important alloying element magnesium in varying concentrations was analyzed using in-situ laser-induced breakdown spectroscopy (LIBS). Magnesium emission shows an exponential dependence on melt temperature that correlates well with the expected partial pressure of magnesium above the aluminum melt. Furthermore, comparison with LIBS measurements on corresponding solid samples supports the conclusion that a significant part of Mg emission from liquid metal samples originates from the vapor phase above the metal surface. Simultaneously, curves of growth measured over four orders of magnitude in Mg concentration reveal a level of self-absorption for liquid aluminum samples that is stronger than for solid aluminum samples having a corresponding Mg concentration, and beyond what is expected from conventional plasma models. The implications for measurements of volatile species in liquid metals in general are discussed.

Directional coupling in long-range dielectric-loaded plasmonic waveguides.

Directional couplers (DCs) based on long-range dielectric-loaded surface plasmon-polariton waveguides (LR-DLSPPWs) operating at telecom wavelengths are studied both numerically and experimentally. The investigated LR-DLSPPWs are formed by ~1.2-µm-high and 1-µm-wide polymer ridges fabricated atop of 15-nm-thick and 500-nm-wide gold stripes supported by a 288-nm-thick Ormoclear polymer deposited on a low-index (n(s) ≈1.34) layer of Cytop. DC structures consisting of sine-shaped S-bends (having an offset of ~10 µm over a distance of ~20 µm) and ~100-µm-long parallel LR-DLSPPWs with a center-to-center separation of 2 µm are characterized using scanning near-field microscopy. The experimentally obtained values of the propagation length (~400 µm), S-bend loss (~4 dB) and coupling length (~100 µm) are found in good agreement with the numerical simulations, indicating a significant potential of LR-DLSPPWs for the realization of various plasmonic components.

On-chip modulation of evanescent illumination and live-cell imaging with polymer waveguides.

Imaging of live cells was carried out using evanescent-wave excitation on a polymer waveguide chip. Integrated waveguide-based interferometric light modulators were fabricated in order to demonstrate on-chip control of excitation light, e.g., for time-lapse fluorescence microscopy. When combined with a sensitive high-resolution imaging system, the integrated waveguide-excitation platform provides an ideal method of near-surface studies of live cells, where photobleaching and/or phototoxicity effects are of critical concern.

Long-range dielectric-loaded surface plasmon polariton waveguides operating at telecommunication wavelengths.

We report on the realization of long-range dielectric-loaded surface plasmon polariton waveguides (LR-DLSPPWs) consisting of straight and bent subwavelength dielectric ridges deposited on thin and narrow metal stripes supported by a dielectric buffer layer covering a low-index substrate. Using imaging with a near-field optical microscope and end-fire coupling with a tapered fiber connected to a tunable laser at telecommunication wavelengths (1425-1545 nm), we demonstrate low-loss (propagation length ∼500 µm) and well-confined (mode width ∼1 µm) LR-DLSPPW mode guiding and determine the propagation and bend loss.

SERS Imaging of Mesenchymal Stromal Cell Differentiation.

Understanding the process of mesenchymal stromal cell (MSC) osteogenic differentiation is essential for a wide range of medical applications. However, these primary cells vary significantly from donor to donor, making it difficult to fully exploit their therapeutic potential. Although osteogenic differentiation has been studied extensively, there is still a shortage of standardized methods for the evaluation of the degree of differentiation. Here, we employ noninvasive surface-enhanced Raman scattering (SERS) for studying such cells, offering a better understanding of cellular processes in situ. We present the long-term differentiation of MSCs on biocompatible gold nanoisland SERS substrates, combining imaging of cells with spectroscopic detection of molecular species and chemical events occurring on the cellular membrane adjacent to the surface of the SERS substrate. We detect multiple signs of bone tissue formation, from an early stage to mature osteoblasts, without labeling. We show that the results correlate very well with classical differentiation-detecting assays, indicating that the SERS imaging technique alone is sufficient to study the progress of osteogenic differentiation of such cells, paving a way toward continuous label-free screening of live cells.

High index contrast polymer waveguide platform for integrated biophotonics.

We present detailed characterization of a unique high-index-contrast integrated optical polymer waveguide platform where the index of the cladding material is closely matched to that of water. Single-mode waveguides designed to operate across a large part of the visible spectrum have been fabricated and waveguide properties, including mode size, bend loss and evanescent coupling have been modeled using effective-index approximation, finite-element and finite-difference time domain methods. Integrated components such as directional couplers for wavelength splitting and ring resonators for refractive-index or temperature sensing have been modeled, fabricated and characterized. The waveguide platform described here is applicable to a wide range of biophotonic applications relying on evanescent-wave sensing or excitation, offering a high level of integration and functionality. The technology is biocompatible and suitable for wafer-level mass production.

Evanescent-wave fluorescence microscopy using symmetric planar waveguides.

We describe a new evanescent-wave fluorescence excitation method, ideally suited for imaging of biological samples. The excitation light propagates in a planar optical waveguide, consisting of a thin waveguide core sandwiched between a sample in an aqueous solution and a polymer with a matching refractive index, forming a symmetric cladding environment. This configuration offers clear advantages over other waveguide-excitation methods, such as superior image quality, wide tunability of the evanescent field penetration depth and compatibility with optical fibers. The method is well suited for cell membrane imaging on cells in culture, including cell-cell and cell-matrix interaction, monitoring of surface binding events and similar applications involving aqueous solutions.

Surface enhanced Raman imaging: periodic arrays and individual metal nanoparticles.

Surface enhanced Raman scattering (SERS) from Rhodamine 6G homogenously adsorbed on both periodic arrays of and individual gold nanoparticles is investigated using high-resolution Raman imaging with polarized excitation. Rectangular 50-nm-high nanoparticles of different sizes chosen to ensure the presence of localized surface plasmon resonances close to the 532-nm excitation wavelength are fabricated with electron-beam lithography on the surface of a smooth gold film and arranged both individually (i.e., placed sufficiently far apart) and in 740-nm-period arrays. Linear reflection spectra and high-resolution Raman images obtained from arrays of nanoparticles are compared revealing good correspondence in the spectral dependences of reflection and local SERS enhancements (measured at the top of nanoparticles). The latter are related to those observed with individual nanoparticles. The results obtained emphasize the importance and quantify the influence of particle dimensions, polarized excitation, collctive resonances and SERS locations.

Gold nanoisland substrates for SERS characterization of cultured cells.

We demonstrate a simple approach for fabricating cell-compatible SERS substrates, using repeated gold deposition and thermal annealing. The substrates exhibit SERS enhancement up to six orders of magnitude and high uniformity. We have carried out Raman imaging of fixed mesenchymal stromal cells cultured directly on the substrates. Results of viability assays confirm that the substrates are highly biocompatible and Raman imaging confirms that cell attachment to the substrates is sufficient to realize significant SERS enhancement of cellular components. Using the SERS substrates as an in vitro sensing platform allowed us to identify multiple characteristic molecular fingerprints of the cells, providing a promising avenue towards non-invasive chemical characterization of biological samples.

Innovative in vitro method to study ventilator induced lung injury.

Mechanical ventilation (MV) is a life-saving therapy for critically ill patients, alleviating the work of breathing and supporting adequate gas exchange. However, MV can cause ventilator induced lung injury (VILI) by baro/volu- and atelectrauma, even lead to acute respiratory distress syndrome (ARDS), and substantially augment mortality. There is a need for specific biomarkers and novel research platforms for VILI/ARDS research to study these detrimental disorders and seek ways to avoid or prevent them. Previous in vitro studies on bronchial epithelium, cultured in air-liquid interface (ALI) conditions, have generally utilized static or constant pressure.  We have developed a Cyclical Pressure ALI Device (CPAD) that enables cyclical stress on ALI cultured human bronchial cells, with the aim of mimicking the effects of MV. Using CPAD we were able to analyze differentially expressed VILI/ARDS and innate immunity associated genes along with increased expression of associated proteins. CPAD provides an easy and accessible way to analyze functional and phenotypic changes that occur during VILI and may provide a platform for future drug testing.

Compact plasmonic variable optical attenuator.

We demonstrate plasmonic nanowire-based thermo-optic variable optical attenuators operating in the 1525-1625 nm wavelength range. The devices have a footprint as low as 1 mm, extinction ratio exceeding 40 dB, driving voltage below 3 V, and full modulation bandwidth of 1 kHz. The polarization dependent loss is shown to be critically dependent on the nanowire geometry but devices with polarization-dependent loss as low as +/-2.5 dB PDL over most of the attenuation range have been fabricated. We propose an even more compact device design to reduce insertion loss to approximately 1 dB.

Microgels and apparatus for PAGE of nucleic acids in one or two dimensions.

We describe a system for horizontal 1D or 2D PAGE comprising an apparatus and microgels. There is no buffer outside the gel, making handling and sample loading easy. Specially designed electrodes on all four sides allow 2D electrophoresis without gel rotation. Electrophoresis is completed within 20 min and sensitivity is in the subnanogram range. The system is temperature controlled for speed, denaturation of nucleic acid molecules and maintaining molecules single-stranded. The system allows characterization of structure, conformation and damage in complex nucleic acid preparations. Besides quick 1D PAGE, 2D applications include characterization of efficiency of complex molecular procedures, checking quality of biosamples and detecting DNA damage in cells and body fluids. The system should also run protein gels.

Near-field imaging of surface plasmon-polariton guiding in band gap structures at telecom wavelengths.

Using a collection near-field microscope, we image interaction of surface plasmon-polaritons (SPPs) excited locally at telecom wavelengths with periodic triangular arrays of gold bumps placed on gold film surfaces. We observe the inhibition of SPP propagation into the arrays within a certain wavelength range depending on their period and orientation, i.e., the band gap (BG) effect, as well as the SPP propagation along bent channels cut through these arrays. Prospects and challenges in realization of compact and efficient SPPBG waveguiding structures are discussed.

Compact Z-add-drop wavelength filters for long-range surface plasmon polaritons.

We design, fabricate and investigate compact Z-add-drop (ZAD) filters for long-range surface plasmon polaritons (LR-SPPs) at telecom wavelengths. The ZAD filter for LR-SPPs consists of two ridge gratings formed by periodic gold thickness modulation at the intersections of three zigzag-crossed gold stripes embedded in polymer. We investigate influence of the grating length and crossing angle on the filter characteristics and demonstrate a 10 masculine- ZAD filter based on 80-microm-long gratings that exhibit a 15-dB dip (centered at ~ 1.55 microm) in transmission of the direct arm along with the corresponding ~ 13-nm-wide transmission peak in the drop arm.

Tetraphenylporphyrin tethered chitosan based carriers for photochemical transfection.

Water-soluble amphiphilic chitosan nanocarriers tethered with the photosensitizer meso-tetraphenylporphyrin were synthesized in a seven-step procedure, starting from 3,6-di-O-tert-butyldimethylsilyl-chitosan and 5-(p-aminophenyl)-10,15,20-triphenylporphyrin. The lipophilic photosensitizer could be introduced in a quantitative and reproducible reaction to give either 0.1 or 0.25 degrees of substitution per glucosamine monomer. Fluorescence and NMR investigations revealed the dynamic structures of the carriers, which formed nanoparticles in aqueous solution with a core of π-stacked photosensitizers. These carriers can then unfold in the lipophilic environment, and the photosensitizer moiety can thus be inserted into the cell membrane. The efficacy of the carriers for photochemical internalization (PCI) mediated gene delivery was evaluated in vitro using the HCT116/LUC human colon carcinoma cell line. The efficacy of transfection was comparable to what could be achieved by the reference compound and current clinical candidate TPCS(2a).