Analytical scale ultrasonic standing wave manipulation of cells and microparticles

The ultrasonic standing-wave manipulation of suspended eukaryotic cells, bacteria and submicron latex or silica particles has been examined here. The different systems, involving plane or tubular ultrasonic transducers and a range of acoustic pathlengths, have been designed to treat suspension volumes of analytical scale i.e. 5 ml to 50 microliters for both sample batch and 'on-line' situations. Frequencies range from 1 to 12 MHz. The influence of secondary cell-cell interaction forces in determining the cell concentration dependence of harvesting efficiency in batch sedimentation systems is considered. Applications of standing wave radiation forces to (1) clarify cell suspensions, (2) enhance particle agglutination immunoassay detection of cells or cellular products and (3) examine and enhance cell-cell interactions in suspension are described.     

Power ultrasonic transducers with extensive radiators for industrial processing

High-power ultrasonics (HPU) is a green emerging technology that offers a great potential for a wide range of industrial processes. Nevertheless such potential have remained restricted during many years to a limited number of applications which reached commercial development. The possible major problem for extending the range of HPU industrial applications has been the lack of power ultrasonic transducers for large-scale application, adapted to the requirements of each specific problem with high efficiency and power capacity.A new family of HPU transducers with extensive radiators have been recently introduced. It comprises a variety of transducer types designed with the radiators adapted to different specific uses in fluids and multi-phase media. Such transducers implement high power capacity, high efficiency and radiation pattern control. In addition, their design incorporate strategies to eliminate or reduce modal interactions produced at high power as a consequence of their nonlinear behaviour. The introduction of such new transducers has significantly contributed to the development at semi-industrial and industrial level of a number of processes in the food and beverage industry, in environment and in manufacturing. This paper deals with the basic structure and main characteristics of such transducers as well as their performance in the developed application processes.     

Molecular masking and unmasking of the paramagnetic effect of iron on the proton spin-lattice (T1) relaxation time in blood and blood clots

The contribution of hemolysis, proteolysis and the paramagnetic effect of iron on the proton spin-lattice (T1) relaxation time in blood was examined. Hemolysis induced by sonication resulted in a significant (10%) increase in the T1 relaxation time of whole blood. Proteolysis in both sonicated and unsonicated whole blood samples eventually yielded T1 values which correlated well with the relaxation times of free iron in plasma or water at concentrations comparable to the concentration of iron in whole blood. It is concluded that proteolysis allows the iron atom to express its paramagnetic effect on water relaxation by gradually destroying the hydrophobic nature of the pocket in which iron resides on the hemoglobin molecule. The contribution of various blood components to the T1 relaxation of whole blood was also studied. The T1 values for packed erythrocytes, intact whole blood, sonicated whole blood, plasma and serum proved to be significantly different from each other. Serum was found to have a significantly (12%) longer T1 relaxation time than plasma. Packed clotted blood in vitro showed no change in the T1 time for at least 13 days while packed erythrocytes showed a shortening of T1 time after 6-8 days.     

Acoustic characterization in whole blood and plasma of site-targeted nanoparticle ultrasound contrast agent for molecular imaging

The ability to enhance specific molecular markers of pathology with ultrasound has been previously demonstrated by our group employing a nanoparticle contrast agent [Lanza et al., Invest. Radiol. 35, 227-234 (2000); Ultrasound Med. Biol. 23, 863-870 (1997)]. One of the advantages of this agent is very low echogenicity in the blood pool that allows increased contrast between the blood pool and the bound, site-targeted agent. We measured acoustic backscatter and attenuation coefficient as a function of the contrast agent concentration, ambient pressure, peak acoustic pressure, and as an effect of duty cycle and wave form shape. Measurements were performed while the nanoparticles were suspended in either whole porcine blood or plasma. The nanoparticles were only detectable when insonified within plasma devoid of red blood cells and were shown to exhibit backscatter levels more than 30 dB below the backscatter from whole blood. Attenuation of nanoparticles in whole porcine blood was not measurably different from that of whole blood alone over a range of concentrations up to eight times the maximum in vivo dose. The resulting data provide upper bounds on blood pool attenuation coefficient and backscatter and will be needed to more precisely define levels of molecular contrast enhancement that may be obtained in vivo.     

Plasma-mediated ablation of biofilm contamination

Ultra-short pulsed laser removal of thin biofilm contamination on different substrates has been conducted via the use of plasma-mediated ablation. The biofilms were formed using sheep whole blood. The ablation was generated using a 1.2 ps ultra-short pulsed laser with wavelength centered at 1552 nm. The blood contamination was transformed into plasma and collected with a vacuum system. The single line ablation features have been measured. The ablation thresholds of blood contamination and bare substrates were determined. It is found that the ablation threshold of the blood contamination is lower than those of the beneath substrates including the glass slide, PDMS, and human dermal tissues. The ablation effects of different laser parameters (pulse overlap rate and pulse energy) were studied and ablation efficiency was measured. Proper ablation parameters were found to efficiently remove contamination with maximum efficiency and without damage to the substrate surface for the current laser system. Complete removal of blood contaminant from the glass substrate surface and freeze-dried dermis tissue surface was demonstrated by the USP laser ablation with repeated area scanning. No obvious thermal damage was found in the decontaminated glass and tissue samples.     

Freeze-drying of mononuclear cells and whole blood of human cord blood

The research on haematopoietic stem cells of human cord blood has become more important recently. People have concentrated on the preservation of cord blood stem cells. At present, cord blood can be preserved at ultra-low temperatures. In this study, we try to preserve cord blood and its constituents by freeze-drying. The experiments on both the mononuclear cell content and the whole blood of human cord blood were carried out respectively. The samples were frozen firstly by different cooling protocols in the presence of PVP, sucrose, and mannitol. Afterwards, they were vacuum-dried at a selected shelf temperature of -30 degree C for the main drying stage, and then vacuum-dried at 15 degree C for the second drying stage. The entire time of the freeze drying was 52 hours. Samples were stored at room temperature for 2 days prior to evaluation. Subsequently, the dried samples were suspended in an isotonic phosphate-buffered saline solution. The recovery of the cells were tested by a haemacytometer, and the highest cell numerical count recovery of MNC was 75.0 percent (SD = 4.1 percent) (P = 0.01), obtained in the protocol of 40 percent PVP + 20 percent sucrose + 10 percent Mannitol. The viability of the nucleated cells measured by PI staining and the ratio of the number of CD34+ to the number of lymphocytes (by the FITC anti-human CD34+ conjugated antibody method) were measured using a flow cytometer (FCM). The protocol of 40 percent PVP + 20 percent sucrose + 10 percent fetal bovine serum had the highest viability of 98.6 percent (SD = 0.7 percent) (P = 0.01). The highest ratio of CD34+ to lymphocytes was 1.2%, and the highest recovery of CD34+ was 68.4 percent (SD = 39.5 percent) (P = 0.05). Comparing the results of the lyophilized MNC subfraction with that of the whole blood, the lyophilization of the isolated MNC was more successful than that of whole blood.     

Investigation of molecular complexes of blood serum by laser correlation spectroscopy and atomic-force microscopy

The progression of pathological processes in the human organism is accompanied by significant changes in a number of molecular parameters of biological fluids. An intensive search for methods appropriate for diagnosing homeostatic disturbances inherent in various pathologies in the preclinical stage has been performed all over the world. Macromolecular fractions of blood serum and blood plasma with typical sizes ranging from 1 to 1000 nm are of particular research interest from the viewpoint of the development of the methods intended for the early detection of oncological and cardiovascular diseases. Considerable possibilities for studying these objects are provided by a combined approach with the use of laser correlation spectroscopy and atomic-force microscopy. The results obtained within the proposed approach have been reported in the present paper. They have demonstrated that this approach holds much promise for composition analysis of blood serum and for further development of the methods employed in the early detection of serious diseases as a whole.     

Resonance tracking and vibration stablilization for high power ultrasonic transducers

Resonant frequency shift and electrical impedance variation are common phenomena in the application of high power ultrasonic transducers, e.g. in focused ultrasound surgery and in cutting. They result in low power efficiency and unstable vibration amplitude. To solve this problem, a driving and measurement system has been developed to track the resonance of high power transducers and to stabilise their vibration velocity. This has the ability to monitor the operating and performance parameters of the ultrasonic transducers in real time. The configuration of the system, with its control algorithm implemented in LabVIEW (National Instruments, Newbury, UK), ensures flexibility to suit different transducers and load conditions. In addition, with different programs, it can be utilised as a high power impedance analyser or an instantaneous electrical power measurement system for frequencies in the MHz range. The effectiveness of this system has been demonstrated in detailed studies. With it, high transducer performance at high power can be achieved and monitored in real time.     

血液的光散射特性

测定了不同血液样品(全血、红细胞、血浆)在可见光的吸收光谱,研究了He-Ne激光经过全血和红细胞的偏振度的变化,得出了人体血液中对可见光的吸收和散射主要来自红细胞的结论.     

Wave guide imaging through Time Domain Topological Energy

Time Domain Topological Energy (TDTE), uses the reflected ultrasonic field recorded by an array of transducers placed on the boundary of the inspected medium. Two numerical determinations (forward and adjoint problems) of the acoustical field inside a reference medium are necessary to compute an image. Topological Energy is defined as a variation of topological sensitivity or gradient and comes from the field of mathematical optimisation. Recent developments for Non-Destructive Testing have shown the analogy with Time Reversal concepts. Time Reversal mirrors have been employed for various applications in a wide number of situations including wave guides where very good re-focalization performances have been obtained with a reduced number of transducers instead of an array of transducers. Moreover recent works have enlightened that the reverberation properties of a wave guide allow to re-focalize using Time Reversal with only one transducer. For TDTE imaging a single transducer placed at one end of a wave guide has been modelled. The boundaries of the wave guide create virtual sources that can be understood as a virtual array of transducers. Numerical and experimental results are presented using TDTE and a single transducer in a wave guide for targets and conditions of increasing complexity.     

血液静态荧光偏振光谱研究

报道了用408 nm的LED偏振光诱导浓度为0.3%的人全血溶液在350800 nm波段的静态荧光偏振光谱,讨论了其光谱的结构特征和红移现象并给出了机理解释;检测了不同浓度全血溶液荧光偏振光谱中在490nm和610 nm附近处两个各主要荧光区的偏振度,实验结果表明,当血液浓度为1.0%时,在两个主要荧光区的偏振度分别为0.4796和0.4344,当浓度增加到8.0%时,相应的偏拓坟墓分别为0.2064和0.0538。研究了偏振度随浓度的变化规律,用能量转移理论分析了在不同荧光区、不同浓度下,血液中各荧光团之间的不同能量转换机理及偏振度的变化现象。研究结果对光诱导生物组织自体荧光诊断技术有一定的参考价值。     

Measurement of elastic nonlinearity using remote laser ultrasonics and CHeap Optical Transducers and dual frequency surface acoustic waves

A nonlinear ultrasonic technique for evaluating material elastic nonlinearity has been developed. It measures the phase modulation of a high frequency (82MHz) surface acoustic wave interacting with a low frequency (1MHz) high amplitude stress inducing surface acoustic wave. A new breed of optical transducers has been developed and used for the generation and detection of the high frequency wave. The CHeap Optical Transducer (CHOT) is an ultrasonic transducer system, optically activated and read by a laser. We show that CHOTs offer advantages over alternative transducers. CHOTs and nonlinear ultrasonics have great potential for aerospace applications. Results measuring changes in ultrasonic velocity corresponding to different stress states of the sample are presented on fused silica and aluminium.     

Structure and function in native and pathological erythrocytes: A quantitative view from the nanoscale

The red blood cells (RBCs) are among the most simple and less expensive cells to purify; for this reason and for their physiological relevance, they have been extensively studied with a variety of techniques. The picture that results is that these cells have several peculiarities including extreme mechanical performances, relatively simple architecture, biological relevance and predictable behavior that make them a perfect laboratory of testing for novel techniques, methodologies and ideas. These include the re-evaluation of old concepts, such as the relationship between structure and function (which is one of the guideline of this report) but considered at the cellular level. The studies reported on this paper, indeed, exploit the full potential of an high resolution quantitative microscopy such as the atomic force microscopy (AFM) to investigate different aspect of the erythrocytes' life, death and interaction with the environment. Indeed, the erythrocytes have a special relationship with the environment that is able to deeply influence their morphology as consequence of alteration of their biochemical or biophysical status. In this context the conditions under which the erythrocytes can be considered as biochemically programmable systems have been investigated by analyzing different environmentally induced alteration of the cell's morphology and comparing the results with naturally occurring pathological morphologies. This class of studies takes great advantage by the additional consideration of the nanomechanical properties of the cells. These latter are particularly important for the cell functionality and are shown to be of practical usefulness to discriminate and partition environmental effects charging different cellular structure (e.g. membrane or membrane-skeleton). Moreover, the development of novel morphological parameter can be important to push the level of investigation on the RBCs' status towards the molecular level. In particular, we describe the introduction and use of the plasma membrane roughness as a morphometric parameter of simple derivation from the AFM images and that results sensitive to the structural integrity of the cells' membrane-skeleton. This offer a remarkable opportunity to investigate the relationship between structure and function in normal and pathological cells by using a morphometric parameter that probes the cell surface at the nanoscale level. At last, a complex but physio-pathologically important phenomenon such as the erythrocytes aging was considered. To properly analyze the many variation that the cells experience during the whole aging path we used all the parameters that the AFM can provides: quantitative imaging, analysis of the membrane roughness and local measure of the nanomechanical properties analyzed together with biochemical parameter such as the ATP content. The picture that emerged is that the aging path is triggered by the ATP intracellular concentration that influence the membrane-skeleton structure and the support exerted on the plasma membrane. The consequences of the membrane-skeleton involvement can be monitored by AFM and showed the occurrence of peculiar morphologies and morphological defects that appear in the very place where the membrane-skeleton contact with the membrane became loose. As a whole, the collected data enable to describe the entire phenomenon as a sequence of morphological intermediates following one another along the aging path.     

用微波ECR等离子体溅射法在蓝宝石(0112)晶面上生长ZnO薄膜的研究

蓝宝石上外延生长ZnO薄膜在表面波和声光器件中有重要的应用．用微波电子回旋共振(ECR)等离子体溅射法在蓝宝石(0112)晶面上外延生长了ZnO薄膜,膜无色透明,并且表面光滑,基片温度为380℃,为探索沉积工艺参数对薄膜结构的影响,用XRD对不同基片温度和沉积速率生长的ZnO薄膜进行了研究．     

Hydrogen passivation of multi—crystalline silicon solar cells

The effects of hydrogen passivation on multi-crystalline silicon (mc-Si) solar cells are reported in this paper. Hydrogen plasma was generated by means of ac glow discharge in a hydrogen atmosphere. Hydrogen passivation was carried out with three different groups of mc-Si solar cells after finishing contacts. The experimental results demonstrated that the photovoltaic performances of the solar cell samples have been improved after hydrogen plasma treatment, with a relative increase in conversion efficiency up to 10.6\%. A calculation modelling has been performed to interpret the experimental results using the model for analysis of microelectronic and photonic structures developed at Pennsylvania State University.     

Influence of size and surface capping on photoluminescence and cytotoxicity of gold nanoparticles

Hydrophilic and homogeneous sub-10 nm blue light-emitting gold nanoparticles (NPs) functionalized with different capping agents have been prepared by simple chemical routes. Structure, average, size, and surface characteristics of these NPs have been widely studied, and the stability of colloidal NP solutions at different pH values has been evaluated. Au NPs show blue PL emission, particularly in the GSH capped NPs, in which the thiol-metal core transference transitions considerably enhance the fluorescent emission. The influence of capping agent and NP size on cytotoxicity and on the fluorescent emission are analyzed and discussed in order to obtain Au NPs with suitable features for biomedical applications. Cytotoxicity of different types of gold NPs has been determined using NPs at high concentrations in both tumor cell lines and primary cells. All NPs used show high biocompatibility with low cytotoxicity even at high concentration, while Au-GSH NPs decrease viability and proliferation of both a tumor cell line and primary lymphocytes.     

Ultrasonic transducers using electron-irradiated vinylidene fluoride-trifluoroethylene copolymers

Single-element, planar transducers have been fabricated using electron-irradiated poly(vinylidene fluoride-trifluoroethylene) 80/20 mol% copolymers with different electron dosage. Electrical field-induced strain response of copolymer film with 100 Mrad dosage has been studied at 5 kHz and the electrostrictive coefficient was calculated. The transmitting response of the air-backing and epoxy-backing transducers was evaluated with the application of high DC bias voltages. Clear ultrasonic amplitudes and high frequency spectrum (>20 MHz) were observed when driven from a standard ultrasonic voltage source through a decoupling circuit. It has also showed that larger generation of ultrasonic waves will be induced under high DC bias field, which is due to the increase of induced d(33) piezoelectric coefficient. Two different polar bias voltages, positive and negative, were applied to the transducers and inverse waveforms were received, which was coincident with the theoretical analysis of the strain response of electrostrictive film.     

Research on inductive transducers for vibration measurement

Recently in Romania the use of high power ultrasonics in electrically conductive solid media has increased. Consequently more severe conditions have been imposed on inductive transducers regarding their sensitivity and selectivity for measuring acoustic field amplitudes. In view of this the authors carried out theoretical and experimental studies on inductive transducers. Parameters that influence the back emf in an idealized physical model, can be particularized for every practical case. By reducing the magnetic reluctance of the air closed field lines, transducers with a sensitivity of about 400 mV/μm have been built. These transducers may possibly be used at higher frequencies. Also, selective transducers for longitudinal and transverse vibrations were made by altering the geometry and position of their constituent elements.     

Atomic force microscopy probing of cell elasticity

Atomic force microscopy (AFM) has recently provided the great progress in the study of micro- and nanostructures including living cells and cell organelles. Modern AFM techniques allow solving a number of problems of cell biomechanics due to simultaneous evaluation of the local mechanical properties and the topography of the living cells at a high spatial resolution and force sensitivity. Particularly, force spectroscopy is used for mapping mechanical properties of a single cell that provides information on cellular structures including cytoskeleton structure.

This entry is aimed to review the recent AFM applications for the study of dynamics and mechanical properties of intact cells associated with different cell events such as locomotion, differentiation and aging, physiological activation and electromotility, as well as cell pathology. Local mechanical characteristics of different cell types including muscle cells, endothelial and epithelial cells, neurons and glial cells, fibroblasts and osteoblasts, blood cells and sensory cells are analyzed in this paper.     

Experimental and finite element modelling studies on single-layer and multi-layer 1-3 piezocomposite transducers

Finite element modelling (FEM) using ATILA code and experimental studies have been carried out on 1-3 piezocomposite transducers. FEM study was initially carried out on a piezocomposite infinite plate and then extended to transducers of finite size. The infinite-plate model results agree well with that of a simple analytical model and experiments. The acoustic performance of multi-layer finite-size piezocomposite transducers was also studied. Transducer stacks were fabricated with different number of layers. The transducer characteristics such as the electrical impedance, the transmitting voltage response (TVR) and the receiving sensitivity (RS) of the 1-3 piezocomposite transducers were evaluated as functions of frequency, ceramic volume fractions and the number of layers. TVR increases and RS decreases with increase in ceramic volume fractions. The model results are found to agree with the experimental data, especially when the number of layers is less.