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Laser interferometry was used to measure sound-induced umbo velocity in the aging mouse middle ear. Velocity reductions of as much as 8 dB were seen as the mice aged. These functional differences suggest a variety of structural changes that may occur in the aging middle ear.
Laser interferometry is proposed for accurate measurements of the external parameters and fixation of the relief of human skull surface. This method creates a detailed three-dimensional computer model of the object of investigation, which can be used in automated systems of personality identification based on investigation of the skull and life-time photos. Further development of the method opens new vistas in automation of the trassological and ballistic identification.
Optical coherence tomography (OCT) is an evolving imaging modality that combines interferometry with low-coherence light to produce high-resolution tissue imaging. Cross-sectional in vivo images were obtained using an OCT device consisting of a Michelson interferometer, 1.3-microm broadband light source, and a handheld fiberoptic imaging probe. Image pixel resolution approached 10 microm. The mucosa of the oral cavity and oropharynx were examined in 41 patients during operative endoscopy. Optical coherence tomographic imaging was combined with endoscopic photography for gross and histologic image correlation. Optical coherence tomographic images of the oral cavity and oropharynx provided microanatomical information on the epithelium, basement membrane (BM), and supporting lamina propria (LP) of the mucosa. Normal microstructures identified in these tissues included an overlying keratin layer, papillae, ducts, glands, and blood vessels. Regions of pathologic features studied included mature scar, granulation tissue, mucous cysts, leukoplakia, and invasive cancer. Optical coherence tomographic imaging showed distinct zones of normal, altered, and ablated tissue microstructures for each pathologic process studied. Abnormal findings were directly compared with regions of normal tissue or conventional histopathologic features when tissue for analysis was available. This study provides a composite series of in vivo OCT images of the oral cavity and oropharynx in a variety of normal regions and pathologic states as well as outline future applications of OCT technology.
Cyanuric chloride activated polyethylene glycol (PEG)-5000 was covalently coupled to murine and human red blood cells (pegylated RBC). Our purpose was to camouflage RBC receptors, which is necessary for parasite invasion, a process essential to sustain parasitemia. Cell electrophoretic mobility analysis (CEM) of pegylated RBC distinguished a new population of cells bearing characteristic CEM. Pegylation of RBC also modified their rheological properties, which were documented by evaluation of cell deformability (based on cell transit time through calibrated micropores) and cell aggregation (as measured by ultrasonic interferometry). Homologous transfusion of pegylated RBC into murine malaria-infected mice had no significant effect on the cerebral malaria death rate in Plasmodium berghei-infected mice, but it reduced the peripheral blood parasitemia by a factor 2 while in Plasmodium yoelii infected mice, the parasitemia was dramatically reduced by a factor of 4. These experiments demonstrate that transfusion of pegylated RBC may inhibit peripheral parasitemia. Cell electrophoresis appears to be a useful tool to allow in vivo detection and to investigate the fate of transfused pegylated RBC.
In the early days of electronic communication-as a result of the limited number of quartz resonators available-frequency adjustment was accomplished by a pencil mark depositing a foreign mass layer on the crystal. In 1959, Sauerbrey showed that the shift in resonance frequency of thickness-shear-mode resonators is proportional to the deposited mass. This was the starting point for the development of a new generation of piezoelectric mass-sensitive devices. However, it was the development of new powerful oscillator circuits that were capable of operating thickness shear mode resonators in fluids that enabled this technique to be introduced into bioanalytic applications. In the last decade adsorption of biomolecules on functionalized surfaces turned in to one of the paramount applications of piezoelectric transducers. These applications include the study of the interaction of DNA and RNA with complementary strands, specific recognition of protein ligands by immobilized receptors, the detection of virus capsids, bacteria, mammalian cells, and last but not least the development of complete immunosensors. Piezoelectric transducers allow a label-free detection of molecules; they are more than mere mass sensors since the sensor response is also influenced by interfacial phenomena, viscoelastic properties of the adhered biomaterial, surface charges of adsorbed molecules, and surface roughness. These new insights have recently been used to investigate the adhesion of cells, liposomes, and proteins onto surfaces, thus allowing the determination of the morphological changes of cells as a response to pharmacological substances and changes in the water content of biopolymers without employing labor-intense techniques. However, the future will show whether the quartz-crystal microbalance will assert itself against established label-free sensor devices such as surface plasmon resonance spectroscopy and interferometry.
We propose an approach for separating nuclear spin isomers with coherent light and illustrate it by numerical calculations using fulvene as a model system. The scheme employs the equivalence of torsion and interchange of equivalent H-atoms in a class of molecules of which fulvene is a simple example. The exchange symmetry couples with the rotational symmetry to produce a spatial distinction between the two photo-excited nuclear spin isomers, and wavepacket interferometry is applied to separate the species.
Using a surface forces apparatus to measure forces, and optical (multiple beam) interferometry to measure surface shapes and separations (to +/-1 A), the normal, viscous, and shear (lubrication) forces between smooth mica surfaces in aqueous hyaluronic acid (HA) solutions were measured. The experimental conditions of loading pressures, pH, and HA concentration were set to closely correspond to physiological human knee-joint conditions. From the force and optical (refractive index) measurements, it was concluded that, like other negatively charged polyelectrolytes, HA does not naturally adsorb on the mica surface which is hydrophilic and weakly negatively charged at physiological conditions: the polymer solution exhibits the bulk viscosity (22.5 +/- 1.5 cP) for films thicker than about 0.4 miccrom of the polymer, whereas for thinner films, the viscosity decreases monotonically toward the value of the pure electrolyte solution (1 cP) as HA is extruded from between the surfaces. This is indicative of a repulsive "depletion" interaction of HA with each mica surface and to a weakly attractive polymer-mediated force between the two surfaces. Thus, free HA in synovial fluid is not expected to act as a good "boundary lubricant." Relaxation measurements on approaching and receding surfaces in HA solutions were also performed, and it is shown that the presence of HA in the bulk solution can improve "hydrodynamic" modes of lubrication, for example, by assuaging the compression stroke. The study includes information that is beneficial to researchers working with biomaterials viscosupplementation devices.
We have investigated the anti-adhesive properties of a newly synthesized fluorescent triblock copolymer containing poly(ethylene oxide). This adsorbs from aqueous solution onto glass that has been rendered hydrophobic. When the polymer-treated surface was exposed to human platelet-rich plasma (PRP) or whole blood at 37 degrees C, platelet adhesion and spreading were prevented. Avid adhesion and rapid platelet spreading occurred along tracks scraped in the adsorbed polymer coating, as seen by video-enhanced interference reflection microscopy. Leukocytes from whole blood are eventually able to adhere to the polymer-treated surface and were seen to remove labeled polymer from their vicinity and accumulate it at the cell body. Interferometry using polystyrene spheres showed that they do not adhere to polymer-coated glass and are unable to approach closer than 70-95 nm. On scraped tracks, beads make molecular contacts with the glass. Because the fully extended solvated (EO)400 arms may extend up to 100 nm from the glass, this suggests that the polymer forms a monolayer with the hydrophilic arms projecting into the water, whereas the hydrophobic (BO)55 segment binds the molecule to the hydrophobic surface. Another tri-bloc copolymer with shorter hydrophilic arms allows particles to approach more closely.
Dentine is a porous hydrated composite structure that forms the major bulk of the human tooth. The aim of this study was to investigate the role of free water on the in-plane, mechanical strain response in dentine structure. A digital moire interferometry was used for this purpose. It was observed from this experiment that structural dentine demonstrated distinct strain gradients in the axial (perpendicular to the dentinal tubules) and lateral (parallel to the dentinal tubules) directions. The hydrated dentine displayed significant increase in strain with stress in the direction perpendicular to the dentinal tubules, and this response was characteristic of a tough material. On the contrary, the dehydrated dentine, which was dehydrated at 24 degrees C, 55% relative humidity for 72 h showed a strain response characteristic of a brittle material. The strains formed in the direction parallel to the dentinal tubules for hydrated dentine were consistent and did not vary much with increase in applied loads. Upon dehydration, the outer dentine experienced higher strains, and the difference between the outer and inner dentine became more conspicuous with increase in loads. This experiment highlights hydration-induced, distinct in-plane strain gradients in the directions perpendicular and parallel to the dentinal tubules in the dentine structure.
This study examined the influences of titanium (Ti) discs with similar surface roughnesses (R(a) values), but with different topographies and chemical compositions, on the adhesion, spreading, and the alkaline phosphatase (ALP) activity of osteoblast-like cells and normal human fibroblasts. The presence of adhesion molecules on the Ti surfaces and their effects on cell activity were also investigated. Two types of Ti discs were prepared. One kind was a mechanically polished Ti disc, and the other type was a disc obtained by the heating of hydroxyapatite (HA) dip-coated Ti. Scanning electron microscopy, optical interferometry, and scanning Auger electron spectroscopy were used to examine the surface morphology, roughness, and chemical composition, respectively, of the superficial Ti layer. The two types of Ti discs had different topographies and chemical compositions, but had similar R(a) values. The cells on both surface types had similar behaviors and ALP activities. A biological evaluation of the surface-modified Ti discs showed that the type I collagen coating was functionally active in terms of cell spreading in both types of Ti discs. In the mechanically polished Ti discs, fibronectin was functionally active in the normal human fibroblasts, but not in the osteoblast-like cells. Cell adhesion was slightly better on the heat-treated HA dip-coated Ti discs, but not on the mechanically polished Ti discs. Type I collagen and fibronectin mediated the adhesion and spreading of osteoblast-like cells through alpha2beta1 integrin and alpha5beta1 integrin, respectively. These results suggest that type I collagen might be a good candidate for the biochemical modification of Ti surfaces, particularly those surfaces obtained by heating of HA dip-coated Ti.
Reflection electron holography is described as a method to observe sub-A surface morphology. Phase shift of a Bragg-reflected electron wave was measured by means of holographic interferometry using an electron microscope equipped with a field emission electron gun and an electron biprism. A short wavelength of high energy electrons is the essential key to the high vertical sensitivity of this method, since geometrical path differences produced by the surface topography are measured in units of wavelengths in interferometrical measuring. Phase shift at a monoatomic step and the displacement field around a dislocation emerging on the surface were observed.
Changes in the cellularity and in the nature of the matrix were studied in the cartilages of the tibial plateau in experimentally induced arthritis in the dog, 7 and 14 days after section of the anterior cruciate ligament. Samples from the different regions of these cartilages were chilled and sectioned in a cryostat, with a variable microtome chuck to allow precise orientation of the specimens. The samples were examined by normal light microscopy, by microscopic interferometry, and by quantitative polarized light microscopy. The orientation of the glycosaminoglycans was assessed by the new "induced birefringence" method. The results indicated that only the region of the medial tibial cartilage that was unprotected by the meniscus was affected, showing increased water content, loss of superficial cells, and a decrease in orientation of the glycosaminoglycans. Whereas the birefringence of the collagen was unaffected, the superficial area that lacked oriented glycosaminoglycans was markedly increased; this may be a useful indicator of early osteoarthritic changes.
The ability of shear strains to inhibit bony ingrowth was investigated by use of a transcortical porous-coated cylindrical plug implant in a functionally isolated turkey ulna model in which the mechanical loading environment could be accurately controlled and rigorously defined. The distribution of ingrowth at the bone-implant interface was quantified following 8 weeks of in vivo loading consisting of 100 seconds per day of a 20 Hz sinusoidal stimulus sufficient to cause a local peak strain of approximately 100 microstrain in the cortex at the bone-implant interface in four turkeys. A nonuniform but repeatable pattern of bony ingrowth, from 33 +/- 6 to 72 +/- 6% (mean +/- SE), was observed. The mechanical environment in the vicinity of the bone-implant interface was calculated using a three-dimensional elastic orthotropic finite element model. The general stress-strain state of the bone as predicted by the finite element model was validated in two additional turkeys using four three-element rosette strain gauges, while high resolution moiré interferometry was used to determine the mechanical state of the region immediately adjacent to the implant itself. Shear strains and stresses were evaluated at the interface and correlated to the pattern of bony ingrowth circumscribing the implant interface. Linear regressions between ingrowth and both shear strain and shear stress were negative, with the values of R = -0.75 and R = -0.78 (p < 0.001), respectively, indicating significant inhibition of ingrowth where shear components were maximal. These results suggest that the minimization of shear stress and strain components is a major determinant in achieving successful ingrowth of bone into a prosthesis.
A surface for the capture of biotin-tagged proteins on matrix-assisted laser desorption/ionisation (MALDI) targets has been investigated. Binding of a poly-L-lysine poly(ethylene glycol)-biotin polymer to glass and gold surfaces has been demonstrated using dual wavelength interferometry. Biotinylated proteins were captured onto this surface using tetrameric neutravidin as a multivalent bridging molecule. Biotin tagging of proteins was achieved by chemical biotinylation or by expressing a protein with a biotinylation consensus sequence in E. coli. The specificity of the surface for biotin-tagged proteins allowed the purification of biotin-tagged glutathione-S-transferase from a bacterial lysate directly onto a MALDI target. Subsequently, the protein was digested on the MALDI target and a protein fingerprint analysis confirmed its presence directly, but no E. coli proteins were detected. Therefore, we conclude that this surface is highly specific for the capture of biotin-labelled proteins and has low non-specific binding properties for non-biotinylated proteins. Furthermore, protein-protein interactions using biotinylated lectins were investigated, and the selective capture of the glycoprotein fetuin with wheat germ agglutinin was demonstrated. Also, immobilised Arachis hypogea agglutinin recognised a minor asialo component of this glycoprotein on the array. The high affinity immobilisation of proteins onto this surface allowed effective desalting procedures to be used which improved the desorption of high molecular weight proteins. Another aspect of this surface is that a highly ordered coupling of the analyte can be achieved which eliminates the search for the sweet spot and allows the creation of densely packed protein microarrays for use in mass spectrometry.
Backscattered electron (BSE) images of bone exhibit graylevel contrast between adjacent lamellae. Mathematical models suggest that interlamellar contrast in BSE images is an artifact due to topographic irregularities. However, little experimental evidence has been published to support these models, and it is not clear whether submicron topographical features will alter BSE graylevels. The goal of this study was to determine the effects of topography on BSE image mean graylevels and graylevel histogram widths using conventional specimen preparation techniques. White-light interferometry and quantitative BSE imaging were used to investigate the relationship between the BSE signal and specimen roughness. Backscattered electron image graylevel histogram widths correlated highly with surface roughness in rough preparations of homogeneous materials. The relationship between BSE histogram width and surface roughness was specimen dependent. Specimen topography coincided with the lamellar patterns within the bone tissue. Diamond micromilling reduced average surface roughness when compared with manual polishing techniques but did not significantly affect BSE graylevel histogram width. The study suggests that topography is a confounding factor in quantitative BSE analysis of bone. However, there is little quantitative difference between low-to-moderate magnification BSE images of bone specimens prepared by conventional polishing or diamond micromilling.
It was found that when an aqueous solution of vinyl monomers is polymerized on a hydrophobic substrate, obvious heterogeneity occurs in the region of the interface. This substrate effect was observed on polytetrafluroethylene (Teflon), polypropylene (PP), polyethylene (PE), polystyrene (PS), and polyvinylchloride (PVC), but not on hydrophilic substrates. Compared with synthesis on hydrophilic surfaces, the surfaces of hydrogels synthesized on a hydrophobic substrate exhibit a larger degree of swelling, a lower surface coefficient of friction and elastic modulus, weaker interfacial adhesion, and reduced interaction with biological cells. This substrate effect has been observed for many types of aqueous monomer solutions. It was found that the above properties are related to the loosely cross-linked architecture, containing some graft-like polymer chains, that is formed on the gel surface when the gel is prepared on a hydrophobic substrate. To understand the mechanism of the substrate effect, two novel optical methods, electric speckle pattern interferometry (ESPI) and real-time laser sheet refraction (RT-LSR), were developed. It was found that oxygen trapped in the composite interface between the monomer solution and rough hydrophobic substrates played an important role in the substrate effect.
Investigating protein-protein interactions by mutational analysis requires practical techniques for quantifying rate constants and equilibrium constants over several orders of magnitude with reasonably high sample throughput. We have employed spectroscopic interferometry for label-free monitoring of the interaction between the cytokine interferon alpha2 (IFNalpha2) and the extracellular domain of its receptor ifnar2 (ifnar2-EC). We implemented a versatile surface chemistry for the glass substrate of this transducer for covalent immobilization of proteins. Affinity capturing with a monoclonal anti-ifnar2-EC antibody (mAb) followed by crosslinking with a second, noncompetitive mAb provided stable, but still reversible, immobilization of ifnar2-EC. We measured kinetics and affinity of numerous of mutants of IFNalpha2 and ifnar2-EC. Dissociation rate constants up to 0.3 s(-1) and association rate constants up to 3 x 10(6) M(-)1 s(-1) were resolved by the system. Dissociation constants down to 200 microM were measured with protein concentrations up to 50 microM without no background signal or nonspecific binding. The instrument detection limit is approximately 10 pm without the need for temperature stabilization or referencing channels. The system proved effective for large-scale mutational analysis involving alanine scanning mutagenesis and double mutant cycles.
We introduce a technique to characterize, in situ, the solid-solid interface formed when a rough metal surface is placed in contact with a smooth mica surface. The technique is based on the use of extended spectral analysis of multiple beam interferometry in conjunction with the surface forces apparatus (SFA). The intensity versus wavelength spectra of light transmitted through the rough metal/mica interface is measured and compared to that transmitted through a smooth metal/mica interface. Interference theory is used to characterize the rough interface based on differences in the measured spectra. Roughness on the order of nanometers can be resolved. The key to our technique is the creation of a perfectly smooth metal/mica interface that is in all respects, except for roughness, identical to the metal/mica interface created in the SFA. We accomplish this by thermal evaporation of a metal film directly onto mica. We demonstrate the effectiveness of the technique by characterizing the structure of silver/mica and gold/mica interfaces. We also show how the technique can be used to investigate the exchange of fluids within the contact region.