Glass Cover Slides Research Articles

Fluorescence imaging assisted by surface modes on dielectric multilayers

We propose a novel approach for wide-field fluorescence imaging based on dielectric multilayers on glass coverslides. It is demonstrated that the fluorescence radiated by emitters in proximity to the multilayer surface can couple to Bloch Surface Waves and subsequently leaks into the glass. The coupled fluorescence is beamed with low divergence according to the Bloch Surface Waves spectral/angular dispersion and then collected with a low-magnification imaging system. Since the coupling between emitter and surface modes is dependent on the surface local morphology, we find a strong spectral deformation of the collected fluorescence spectrum depending on the corresponding surface relieves imaged.

Selective functionalization of patterned glass surfaces.

Tailored writing and specific positioning of molecules on nanostructures is a key step for creating functional materials and nano-optical devices, or interfaces for synthetic machines in various applications. We present a novel approach for the selective functionalization of patterned glass surfaces with functional probes of any nature. The presented strategy is optimized for imaging fluorophore labeled nanostructures for (single-molecule) fluorescence microscopy. The first step in the protocol is coating a glass surface, here a microscope cover slide, with a 60 nm thick diamond-like carbon film. Subsequently, the pattern is defined by either writing silicon oxide on the coating with a focused electron beam, or by etching the coating with a focused ion beam to expose the glass surface. Finally, the pattern is silanized and functionalized. We demonstrate the selective binding of organic fluorophores and imaging with high contrast, especially in total-internal-reflection mode. The presented approach is flexible and combines bottom-up assembly with high-resolution lithography on glass cover slides to precisely position and image functional molecules of any type.

Autocalibrating Stokes polarimeter for materials characterization

The design and construction of an ellipsometer based on a fixed-wavelength rotating-retarder Stokes polarimeter is described. Details are provided for an automated calibration scheme that provides two advantages for its operation. The first allows the phase of the lock-in amplifiers to be set based on the raw data, without a known calibration sample. The second illustrates that the relative amplitude of the acquired signals may also be calibrated in a similar manner. As an illustration, the refractive index and thickness of a glass cover slide are determined over a range of incident angles.

Self-assembly of microparticles in stable ring structures in an optical trap

Micro-particle self assembly under the influence of optical forces produced by higher order optical beams or by projection of a hologram into the trapping volume is well known. In this paper, we report the spontaneous formation of a ring of identical microspheres (each with diameter 1.1 $\mu$m) in conventional single beam optical tweezers with a usual TEM$_{00}$ Gaussian beam coupled into a sample chamber having standing wave geometry with a cover slip and glass slide. The effects of different experimental parameters on the ring formation are studied extensively. The experimental observations are backed by theoretical simulations based on a plane wave decomposition of the forward and backward propagating Gaussian beams. The ring patterns are shown to be caused due to geomterical aberrations produced by focusing the Gaussian beam using a high numerical aperture microscope objective into stratified media. It is found that the thickness of the stratified media and the standing wave geometry itself play a critical role in formation of stable ring structures. These structures could be used in the study of optical binding, as well as biological interactions between cells in an optical trap.

Matrine Inhibits Disturbed Flow–Enhanced Migration via Downregulation of ERK1/2–MLCK Signaling Vascular Smooth Muscle Cells

To investigate the effects of matrine on the vascular smooth muscle cell (VSMC) migration modulated by disturbed flow and their underlying molecular mechanisms in vitro. Isolated rat aortic VSMCs were grown to confluence on 20- × 80-mm fibronectin-coated glass cover slides, and then, denuded zones were made at the position calculated to be the oscillating flow-reattachment zone and also in the downstream laminar flow region. VSMCs were treated with different doses of matrine (0, 10, 20, 30, and 40 mg/L), or PD98059 (30 μM), ML-7 (10 μM) combined with matrine (40 mg/L) for 30 minutes before and during the experiments. Then, the wounded monolayers were kept under static conditions or were subjected to laminar or disturbed flow for 21 hours or 10 hours. The VSMC migration was assessed by microscopic images. The extracellular signal-regulated kinase 1/2 (ERK1/2) and myosin light chain kinase (MLCK) proteins were determined by Western blot. Disturbed flow significantly increased phosphorylation of ERK1/2. Selective inhibition of ERK1/2 phosphorylation by inhibitor PD98059 and matrine significantly suppressed VSMC migration under disturbed flow. Disturbed flow significantly enhanced phosphorylation of MLCK, whereas both matrine and PD98059 inhibited the phosphorylation of MLCK under disturbed flow. The complete inhibition of MLCK phosphorylation using the selective MLCK inhibitor ML-7 significantly inhibited VSMC migration under disturbed flow. Matrine inhibits VSMC migration under disturbed flow, in part, by downregulation of ERK1/2-MLCK signaling pathway.

Hydrophobicity and Biofilm Growth ofE-coliRP437, MG1655 and B/r

ABSTRACTTwo strains ofE-ColiK-12, viz, RP437, MG1655 and B/r (E. coliB derivative, not a K-12 strain) were grown on various surfaces to study bacterial adhesion and subsequent biofilm formation. We observed biofilm and large colonies on cover slides, beads made of soda lime or borosilicate glasses, on plasma treated PDMS (Polydimethylsiloxane), on Tissue Culture (TC) polystyrene, and observed some clusters on plasma treated ZnTi cover slide; but no evidence of biofilm on untreated-PDMS and ZnTi glass cover slides. From contact angle measurements, we conclude that the hydrophobic nature of untreated PDMS prevent bacterial adhesion for these three strains.

Oriented Poly(3-hexylthiophene) Nanofibril with the π−π Stacking Growth Direction by Solvent Directional Evaporation

In this article, the uniaxial alignment of poly(3-hexylthiophene) (P3HT) nanofibrils with a π-π stacking growth direction in which P3HT chains adopt a flat-on conformation was obtained by solvent directional evaporation using a glass cover slide and a poly(dimethylsiloxane) (PDMS) sheet to press the P3HT film in a carbon disulfide (CS(2)) atmosphere. By controlling the CS(2) vapor pressure during the film-forming process, we got a well-oriented P3HT film whose order parameter reached as high as 0.97. The orientation of the film was induced by the crystallization nucleation of P3HT and the directional evaporation of the solvent. Under a CS(2) vapor atmosphere, P3HT crystals preferred to adopt the form II modification, which started by nucleation. Owing to the solvent directional evaporation from the center to the margin, P3HT at the center of the sample would precipitate first to induce nucleation. Then the peripheral P3HT would directly diffuse, precipitate, and then adhere to the nucleus to form the uniaxial alignment of P3HT nanofibrils along the direction of solvent evaporation. Furthermore, in the P3HT nanofibrils, the π-π stacking direction of P3HT lamellae was parallel to the crystal growth direction, which would provide an effective path for charge transport.

Peroxiredoxin II Restrains DNA Damage-induced Death in Cancer Cells by Positively Regulating JNK-dependent DNA Repair

The 2-Cys peroxiredoxins (Prx) belong to a family of antioxidant enzymes that detoxify reactive oxygen and nitrogen species and are distributed throughout the intracellular and extracellular compartments. However, the presence and role of 2-Cys Prxs in the nucleus have not been studied. This study demonstrates that the PrxII located in the nucleus protects cancer cells from DNA damage-induced cell death. Although the two cytosolic 2-Cys Prxs, PrxI and PrxII, were found in the nucleus, only PrxII knockdown selectively and markedly increased cell death in the cancer cells treated with DNA-damaging agents. The increased death was completely reverted by the nuclearly targeted expression of PrxII in an activity-independent manner. Furthermore, the antioxidant butylated hydroxyanisole did not influence the etoposide-induced cell death. Mechanistically, the knockdown of Prx II expression impaired the DNA repair process by reducing the activation of the JNK/c-Jun pathway. These results suggest that PrxII is likely to be attributed to a tumor survival factor positively regulating JNK-dependent DNA repair with its inhibition possibly sensitizing cancer cells to chemotherapeutic agents.

Synthesis and electro-polymerisation of a novel heteropentalene mesomeric betaine: preparation of a novel low band-gap conjugated polymer

The 3,4-ethylenedioxythiophene (EDOT)-containing heteropentalene mesomeric betaine 9 has been prepared and electro-polymerised. When coated onto an indium tin oxide (ITO) covered glass slide the optical band-gap of the polymer was determined as 1.75 eV.

Direct laser-writing of dielectric-loaded surface plasmon–polariton waveguides for the visible and near infrared

We demonstrate flexible and low-cost fabrication of dielectric-loaded surface plasmon–polariton waveguides. The waveguide structures are fabricated by two-photon polymerization of commercially available, spin-coatable epoxy-based UV-lithographic resist on a metal covered glass slide. The excitation and guiding properties of the plasmonic waveguides are investigated in the far-field at a wavelength of 632.8 nm by imaging the leakage radiation from the waveguide modes. The optimum bending radius for right angle bends is measured to 6 μm providing a transmission of up to 70%. The functionality of more complex Y-splitters is demonstrated.

Formation and hydrogen production of photosynthetic bacterial biofilm under various illumination conditions

The application of immobilized-cell technology in photobioreactor for hydrogen production could offer improvements in photo-hydrogen production rate and light utilization efficiency. Indigenous Rhodopseudomonas palustris CQK 01 was attached to the surface of a cover glass slide in a flat-panel photobioreactor, to form biofilm under illumination with a range of intensities and wavelengths. The morphology and structure of mature photosynthetic bacterial (PSB) biofilm were determined to elucidate the relationship between biofilm formation and hydrogen production performance. The effects of operation conditions on hydrogen production performance of the biofilms formed under various illumination conditions were experimentally investigated. The results showed that illumination wavelength and intensity substantially influenced the morphology and structure of the biofilm, and the hydrogen production performance of mature biofilm varied significantly with the illumination conditions that were used for biofilm formation. Biofilm formed under 590 nm and 5000 lx illumination showed the highest hydrogen production performance.

A micro-channel-well system for culture and differentiation of embryonic stem cells on different types of substrate

We have developed a combined micro-channel and micro-well system for easy cell loading, culture and post-culture operation on a chip. To demonstrate the reliability of the system, on chip cell culture and differentiation were performed with different types of substrates made of culture dish, glass cover slide and polydimethylsiloaxe (PDMS). As expected, mouse embryo fibroblasts (MEF) showed different adhesion and growth rate on different substrates. When embryonic stem (ES) cells were co-cultured with MEFs, the formation of ES colonies is efficient on both glass and Petri dish, although PDMS could also be used. Finally, ES cell differentiation with neuron growth factors was performed on different substrates, showing clear advantages of using culture Petri dish over both glass and PDMS.

Mode-selective excitation of laser-written dielectric-loaded surface plasmon polariton waveguides

We experimentally demonstrate mode-selective excitation of laser-written dielectric-loaded surface plasmon polariton waveguides. The waveguide structures are fabricated by two-photon polymerization of spin-coatable epoxy-based resist on a metal covered glass slide. The waveguides are excited by illuminating the waveguide ends with a strongly focused Gaussian laser beam with a wavelength of 632.8 nm and adjustable polarization. The guiding properties are investigated in the far field by imaging the leakage radiation of the waveguide modes from the metal-substrate interface. It is observed that different TE and TM waveguide modes can be selectively excited with high precision. Simultaneous excitation of the plasmonic TM00 and TM01 modes leads to mode beating and oscillation of the guided intensity between the two sides of the waveguides. In Y-splitters, this simultaneous excitation of the first two TM modes can be used to control the splitting ratio at the exit ports of the splitter.

Real time in vitro measurement of oxygen uptake rates for HEPG2 liver cells encapsulated in alginate matrices

Quantitative non-invasive measurement of critical physiological parameters is necessary to assess the functionality and applicability of tissue engineered matrices. Advancements in fiber optic sensors have made it possible for measuring parameters such as oxygen, glucose, and aminoacids necessary for viable tissue growth. In this study, we have devised an experimental protocol to measure in real time, the oxygen uptake rate (OUR) values for a selected liver cell line (HEPG2) when grown (a) on cover glass slides, and (b) encapsulated within alginate based hydrogel matrices. For both cases, the oxygen uptake rates of HEPG2 cells at selected time points varied in close co-relation with cell proliferation and metabolic activity during the 7-day culture period. This investigation concludes that OUR can be used as an indicative parameter to assess the metabolic activity of cells encapsulated within a matrix. The study also presents a fiber optic sensing technology as a non-invasive diagnostic tool to monitor cell behavior and activity.

Iron-mediated Aggregation and a Localized Structural Change Characterize Ferritin from a Mutant Light Chain Polypeptide That Causes Neurodegeneration

Nucleotide insertions in the ferritin light chain (FTL) polypeptide gene cause hereditary ferritinopathy, a neurodegenerative disease characterized by abnormal accumulation of ferritin and iron in the central nervous system. Here we describe for the first time the protein structure and iron storage function of the FTL mutant p.Phe167SerfsX26 (MT-FTL), which has a C terminus altered in sequence and extended in length. MT-FTL polypeptides assembled spontaneously into soluble, spherical 24-mers that were ultrastructurally indistinguishable from those of the wild type. Far-UV CD showed a decrease in α-helical content, and 8-anilino-1-naphthalenesulfonate fluorescence revealed the appearance of hydrophobic binding sites. Near-UV CD and proteolysis studies suggested little or no structural alteration outside of the C-terminal region. In contrast to wild type, MT-FTL homopolymers precipitated at much lower iron loading, had a diminished capacity to incorporate iron, and were less thermostable. However, precipitation was significantly reversed by addition of iron chelators both in vitro and in vivo. Our results reveal substantial protein conformational changes localized at the 4-fold pore of MT-FTL homopolymers and imply that the C terminus of the MT-FTL polypeptide plays an important role in ferritin solubility, stability, and iron management. We propose that the protrusion of some portion of the C terminus above the spherical shell allows it to cross-link with other mutant polypeptides through iron bridging, leading to enhanced mutant precipitation by iron. Our data suggest that hereditary ferritinopathy pathogenesis is likely to result from a combination of reduction in iron storage function and enhanced toxicity associated with iron-induced ferritin aggregates.

Slipping friction of an optically and magnetically manipulated microsphere rolling at a glass-water interface

The motion of submerged magnetic microspheres rolling at a glass-water interface has been studied using magnetic rotation and optical tweezers combined with bright field microscopy particle tracking techniques. Individual microspheres of varying surface roughness were magnetically rotated both in and out of an optical trap to induce rolling along either plain glass cover slides or glass cover slides functionalized with polyethylene glycol. It has been observed that the manipulated microspheres exhibited nonlinear dynamic rolling-while-slipping motion characterized by two motional regimes: At low rotational frequencies, the speed of microspheres free rolling along the surface increased proportionately with magnetic rotation rate; however, a further increase in the rotation frequency beyond a certain threshold revealed a sharp transition to a motion in which the microspheres slipped with respect to the external magnetic field, resulting in decreased rolling speeds. The effects of surface-microsphere interactions on the position of this threshold frequency are posed and investigated. Similar experiments with microspheres rolling while slipping in an optical trap showed congruent results.

Polarization Modulation Infrared Reflection Absorption Spectroscopy Investigations of Thin Silica Films Deposited on Gold. 2. Structural Analysis of a 1,2-Dimyristoyl-sn-glycero-3-phosphocholine Bilayer

In this paper we report on the structural analysis of bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) using polarization modulation infrared reflection absorption spectroscopy (PM IRRAS). The lipid bilayers were formed on SiO2|Au and Au surfaces using the Langmuir-Blodgett and Langmuir-Schaeffer techniques. As we showed in part 1 (Zawisza, I.; Wittstock, G.; Boukherroub, R.; Szunertis, S. Langmuir 2007, 23, 9303-9309), SiO2 layers of 7 nm thickness, synthesized by plasma-enhanced chemical vapor deposition on 200 nm thick gold covered glass slides, allow PM IRRAS investigations. Only minor changes in the order and structure of the lipid bilayer are observed when deposited on SiO2|Au and Au surfaces. The choline moiety in the leaflet directed toward the SiO2 surface exists in trans conformation and shows a tilt of 28 degrees with the surface normal of the CN bond. On the silica surface in the second leaflet directed toward air and in two layers deposited on the Au surface, trans and gauche isomers of the choline moiety are present and the tilt of the CN bond increases to 55 degrees with respect to the surface normal. The order and molecular orientation in the DMPC bilayers on SiO2 and Au surfaces are not affected by time. The analysis of the phosphate stretching mode on the Au surface shows slight dehydration of this group and reorientation of the phosphate moiety.

WO3-x Nanorods Synthesized on a Thermal Hot Plate

A simple thermal vapor deposition technique to synthesize crystalline tungsten oxide nanorods in ambient condition is presented. Using a commercial thermal hot plate, a pure 99.9% tungsten foil is heated at 485 ± 5 °C under ambient conditions with a piece of 150 μm thick glass cover slide pressing on the tungsten foil. Single crystalline WO3-x nanorods, with a preferential growth axis in the [001] direction, are found to deposit on the cover slide facing the heated tungsten foil. The structure, morphology, and composition of the WO3-x nanorods were characterized using the scanning electron microscopy, transmission electron microscopy, energy dispersive spectrometry, Raman spectroscopy, and X-ray diffraction. In addition, mechanical properties of the as-synthesized nanorods were investigated by employing the three-point bend test using an atomic force microscope. The elastic modulus of the nanorods was found to be in the 10−110 GPa range, and it increases with decreasing diameter of the nanorods. The tungsten oxide nanorod was also found to be a good field emitter with a field enhancement factor estimated to be 9.8 × 104 cm-1.

PM IRRAS Investigation of Thin Silica Films Deposited on Gold. Part 1. Theory and Proof of Concept

Polarization modulation infrared reflection absorption spectroscopy (PM IRRAS) was successfully used for the first time to characterize an optically transparent thin oxide film. SiO2 layers of 7 nm thickness were synthesized by plasma enhanced chemical vapor deposition (PECVD) on 200 nm thick gold covered glass slides. Despite the fact that silica is transparent and absorptive to IR radiation when deposited in the form of thin films on a gold surface, it preserves the high metallic reflectivity for the IR light. At grazing angles of incidence of the IR beam, the enhancement of the normal component of the electric field at the interface is comparable to that of Au alone. In addition, the analysis of the structure of a 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid monolayer deposited using the Langmuir-Blodgett technique is demonstrated.

Identification of Glioma Neovascularization-related Proteins by Using MALDI-FTMS and Nano-LC Fractionation to Microdissected Tumor Vessels

The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.