具有三维聚焦之微型流式细胞/颗粒计数仪

本研究提出一种新式的微型细胞颗粒计数器,此微型细胞颗粒计数器结合二维水力聚焦及微文件流结构(micro-weir structure)于微管道中,可进行高精确度及均一性的细胞或颗粒计数,本研究是利用简单的等向性(isotropie)的湿式蚀刻技术来制作微文件流结构于玻璃基材上,而此微型细胞颗粒计数器的主要构成的组件包含二维边鞘流的聚焦结构,其主要功能为将细胞或颗粒聚焦于x-Y平面上,而微文件流结构的目的在于将细胞或颗粒在z方向的筛选,最后利用雷射诱导荧光系统将细胞或颗粒侦测出.在实验与数值分析的结果,显示此微型细胞颗粒计数器确实可进行高精确度及均一性的细胞或颗粒的计数,并且可以提供一个微型化的生物分析系统.     

Hydrodynamic focusing of a particle flux

Based on numerical integration of the equations of mechanics of multiphase media, an effect of focusing of a particle flux generated by a source located on the upper wall of a closed vessel has been revealed and investigated.Notation t time - x, y Cartesian coordinates - U₁ (u ₁, v₁),P, ₁ velocity, pressure, and density of the gas - ₂,U ₂ mean density and velocity of the dispersed phase - V _k , r _k velocity and radius vector of a macroparticle - g gravitational acceleration - e(0–1) gravity force vector - dynamic viscosity of the gas - f friction force of particles in the gas - Eu, Re Euler and Reynolds numbers - dimensionless time of the high-rate relaxation of particles Élektrogorsk Research Center, Russia; Institute of Mechanics and Biomechanics, Sofia, Bulgaria; Institute for Problems in Mechanics, Russian Academy of Sciences, Moscow. Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 68, No. 3, pp. 355–360, May–June, 1995.     

Thermostatic and dynamic performance of an ultrasonic density probe

The thermally static and dynamic performance of an ultrasonic density probe for liquids is investigated in the density range of 750 to 1300 kg/m³ at temperature ranging from 0 to 40°C. The single transducer probe uses a pulse echo technique to obtain the characteristic acoustic impedance of the liquid and, subsequently, the speed of sound through the liquid to obtain the density of the liquid. Variations in the initial sound amplitude are addressed by the design of a layered two material probe. It is shown that it is possible to obtain an accuracy of 0.4% in the experiments carried out. For changing temperature, the probe exhibits large errors because of problems in estimating the temperatures in certain regions of the probe     

Analytical electric field and sensitivity analysis for two microfluidic impedance cytometer designs

Microfabricated impedance cytometers have been developed to measure the electrical impedance of single biological particles at high speed. A general approach to analytically solve the electric field distributions for two different designs of cytometers: parallel facing electrodes and coplanar electrodes, using the Schwarz-Christoffel Mapping method is presented. Compared to previous analytical solutions, our derivations are more systematic and solutions are more straightforward. The solutions have been validated by comparison with numerical simulations performed using the finite element method. The influences on the electric field distribution due to the variations in the geometry of the devices have been discussed. A simple method is used to determine the impedance sensitivity of the system and to compare the two electrode designs. For identical geometrical parameters, we conclude that the parallel electrodes design is more sensitive than the coplanar electrodes.     

Microchip flow cytometry using electrokinetic focusing

Flow cytometry of fluorescently labeled and unlabeled latex particles is demonstrated on a microfabricated device. The latex particles were detected and counted using laser light scattering and fluorescence coincidence measurements. Sample confinement was accomplished using electrokinetic focusing at a cross intersection, and detection occurred 50 μm downstream from the intersection. Particles with diameters of 1 and 2 μm were analyzed and distinguished from each other based on their light scattering intensity and fluorescence. A maximum sample throughput of 34 particles/s was achieved. Sample mixtures with varying proportions of fluorescently labeled and unlabeled particles were also analyzed and found to be within experimental error of the expected ratios.     

Choosing ultrasonic flow transducers

We consider flowmeters using piezoelectric transducers attached to pipes; these devices are characterized by high error and low sensitivity. Translated from Izmeritel'naya Tekhnika, No. 10, pp. 18–20, October, 1997.     

The effects of focusing and refraction on Gaussian ultrasonic beams

A scalar theory of the propagation of Gaussian ultrasonic beams through lenses and interfaces is presented. For radiation into a fluid, the Fresnel approximation is employed to derive the laws of propagation of Guassian beams (previously employed in the analysis of coherent optical systems). These are then generalized to situations commonly found in nondestructive evaluation by treating the effects of propagation through lenses and through curved interfaces at oblique incidence. A numerical example illustrates the ease with which insight into diffraction phenomena for complex geometries can be gained by this approach. The limitations imposed on the theory by aberrations and the scalar assumption are discussed, and the relationship of the Gaussian theory to the radiation of piston transducers is explored.     

Microfluidic flow cytometer for quantifying photobleaching of fluorescent proteins in cells

Traditional flow cytometers are capable of rapid cellular assays on the basis of fluorescence intensity and light scatter. Microfluidic flow cytometers have largely followed the same path of technological development as their traditional counterparts; however, the significantly smaller transport distance and resulting lower cell speeds in microchannels provides for the opportunity to detect novel spectroscopic signatures based on multiple, nontemporally coincident excitation beams. Here, we characterize the design and operation of a cytometer with a three-beam, probe/bleach/probe geometry, employing HeLa suspension cells expressing fluorescent proteins. The data collection rate exceeds 20 cells/s under a range of beam intensities (5 kW to 179 kW/cm(2)). The measured percent photobleaching (ratio of fluorescence intensities excited by the first and third beams: S(beam3)/S(beam1)) partially resolves a mixture of four red fluorescent proteins in mixed samples. Photokinetic simulations are presented and demonstrate that the percent photobleaching reflects a combination of the reversible and irreversible photobleaching kinetics. By introducing a photobleaching optical signature, which complements traditional fluorescence intensity-based detection, this method adds another dimension to multichannel fluorescence cytometry and provides a means for flow-cytometry-based screening of directed libraries of fluorescent protein photobleaching.     

Analysis of an ultrasonic method of measuring the flow rate of materials

A metrological analysis of an ultrasonic method of measuring the flow rate of materials is presented. Translated from Izmeritel'naya Tekhnika, No. 1, pp. 34–36, January, 2000.     

基于相关性延迟分析的SAFT算法

针对超声经典合成孔径聚焦(SAFT)算法需对每点进行延迟聚焦、运算量大等问题,提出基于相关性延迟分析的合成孔径聚焦算法,并建立数学模型,采用Matlab分别对经典SAFT算法和基于相关性延迟分析的SAFT算法进行仿真分析。结果表明:基于相关性延迟分析的SAFT算法具有图像质量优、分辨率高、实时性好等特点。     

Measurement of scattering properties of individual particles with a scanning flow cytometer

A hydrofocusing head with an optical cuvette has been developed for the flow cytometer to generate complete scatter patterns of single particles at scattering angles ranging from 10° to 120°. The scatter signal has been measured as a function of the angle (a flying indicatrix) by the use of particle motion within a scanning system of the flow cytometer by the use of a single photomultiplier. Scattering data measured with the flow cytometer have been compared with those calculated from Mie theory for latex particles. A calculation algorithm has been used to estimate the size and the refractive index of spherical particles from the scattering data measured.     

Multinode acoustic focusing for parallel flow cytometry

Flow cytometry can simultaneously measure and analyze multiple properties of single cells or particles with high sensitivity and precision. Yet, conventional flow cytometers have fundamental limitations with regards to analyzing particles larger than about 70 μm, analyzing at flow rates greater than a few hundred microliters per minute, and providing analysis rates greater than 50,000 per second. To overcome these limits, we have developed multinode acoustic focusing flow cells that can position particles (as small as a red blood cell and as large as 107 μm in diameter) into as many as 37 parallel flow streams. We demonstrate the potential of such flow cells for the development of high throughput, parallel flow cytometers by precision focusing of flow cytometry alignment microspheres, red blood cells, and the analysis of a CD4+ cellular immunophenotyping assay. This approach will have significant impact toward the creation of high throughput flow cytometers for rare cell detection applications (e.g., circulating tumor cells), applications requiring large particle analysis, and high volume flow cytometry.     

Analytical calculation of the longitudinal electric field resulting from the tight focusing of an ultrafast transverse-magnetic laser beam

A purely time-domain approach is proposed for the propagation of vectorial ultrafast beams in free space beyond the paraxial and the slowly varying envelope approximations. As an example of application of this method, we describe in detail the vectorial properties of an ultrafast tightly focused transverse-magnetic (TM(01)) beam, where special attention is given to the longitudinal electric field component. We show that for spot sizes at the waist comparable to the wavelength, the beam diverges more rapidly than expected from paraxial theory. A consequence of this phenomenon is a faster decrease of the amplitude of the longitudinal field away from the waist and a faster evolution of the axial Gouy phase shift in the vicinity of the focus. It has been observed that the phase of the beam has an overall variation of 2pi from z=-infinity to infinity, independent of the beam spot size at the waist and pulse duration.     

The velocity and attenuation of outgoing surface acoustic waves measured using an ultrasonic microscope with two focusing transducers

A new method for measuring the local parameters of outgoing surface acoustic waves (OSAWs) is proposed. The technique employs a system of two focusing transducers, with the output signal taken from one of these scanning over the sample surface. In comparison with the conventional method with a single transducer moving in the direction normal to the sample surface, the proposed technique is capable of conducting measurements in a greater range of OSAW velocities and is less sensitive to parameters of the immersion liquid. The new method was experimentally verified by measuring systems with known properties.     

Analytical representation of transient ultrasonic phased-array near- and far-fields

The present contribution provides a method for the derivation of analytical near- and far-field expressions for the pressure step-function response of nonuniformly excited ultrasonic transducers modeled as pistons of arbitrary shape in an infinite rigid baffle. Explicit inversion of the Laplace transformed ultrasonic field using Cagniard's idea results in elementary functions or definite integrals of elementary functions, depending on the assumed nonuniform aperture distribution. The basic signal distortion is illustrated in terms of three-dimensional space-time plots of the pressure unit step-function response of a rectangular uniform aperture. The corresponding results for a phased-array transducer are also given. In addition, we investigate the pressure profile on the axis perpendicular to a uniform rectangular piston and its horizontal radiation pattern for different bandwidths of the exciting impulse.     

Model of a two-dimensionally focusing ultrasonic X-ray lens based on Bragg-Fresnel optics

A model is proposed for a two-dimensionally focusing x-ray lens, whose operation is based on the principle of Bragg-Fresnel optics. It is shown that the lens has a large aperture. Pis’ma Zh. Tekh. Fiz. 23, 40–43 (March 26, 1997)     

Design and performance testing of an ultrasonic linear motor with dual piezoelectric actuators

In this work, design and performance testing of an ultrasonic linear motor with dual piezoelectric actuator patches are studied. The motor system consists of a linear stator, a pre-load weight, and two piezoelectric actuator patches. The piezoelectric actuators are bonded with the linear elastic stator at specific locations. The stator generates propagating waves when the piezoelectric actuators are subjected to harmonic excitations. Vibration characteristics of the linear stator are analyzed and compared with finite element and experimental results. The analytical, finite element, and experimental results show agreement. In the experiments, performance of the ultrasonic linear motor is tested. Relationships between velocity and pre-load weight, velocity and applied voltage, driving force and applied voltage, and velocity and driving force are reported. The design of the dual piezoelectric actuators yields a simpler structure with a smaller number of actuators and lower stator stiffness compared with a conventional design of an ultrasonic linear motor with fully laminated piezoelectric actuators.     

Estimating particle concentration using passive ultrasonic measurement of impact vibrations

A method of obtaining particle size and concentration information, from ultrasonic transducer measurements of particle-wall interactions in a particle laden fluid, is presented. A mathematical model of the flexural vibrations of the vessel wall due to the particle impact is constructed. The key component of this model is the derivation of an analytic expression for the impact force amplitude. An analytic expression for the power spectrum is then obtained that shows its explicit dependency on the system parameters. This spectrum is then integrated over a specific frequency range and a comparison with experimental results is reported.