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颗粒与细胞的超声操控。细胞的超声分离。

Ultrasonic manipulation of particles and cells. Ultrasonic separation of cells.

作者信息

Coakley W T, Whitworth G, Grundy M A, Gould R K, Allman R

机构信息

School of Pure and Applied Biology, University of Wales College of Cardiff, UK.

出版信息

Bioseparation. 1994 Apr;4(2):73-83.

PMID:7765041
Abstract

Cells or particles suspended in a sonic standing wave field experience forces which concentrate them at positions separated by half a wavelength. The aims of the study were: (1) To optimise conditions and test theoretical predictions for ultrasonic concentration and separation of particles or cells. (2) To investigate the scale-up of experimental systems. (3) To establish the maximum acoustic pressure to which a suspension might be exposed without inducing order-disrupting cavitation. (4) To compare the efficiencies of techniques for harvesting concentrated particles. The primary outcomes were: (1) To design of an acoustic pressure distribution within cylindrical containers which led to uniformly repeating sound pressure patterns throughout the containers in the standing wave mode, concentrated suspended eukaryotic cells or latex beads in clumps on the axis of wide containers, and provided uniform response of all particle clumps to acoustic harvesting regimes. Theory for the behaviour (e.g. movement to different preferred sites) of particles as a function of specific gravity and compressibility in containers of different lateral dimensions was extended and was confirmed experimentally. Convective streaming in the container was identified as a variable requiring control in the manipulation of particles of 1 micron or smaller size. (2) Consideration of scale-up from the model 10 ml volume led to the conclusion that flow systems in intermediate volume containers have more promise than scaled up batch systems. (3) The maximum acoustic pressures applicable to a suspension without inducing order-disrupting cavitation or excessive conductive streaming at 1 MHz and 3 MHz induce a force equivalent to a centrifugal field of about 10(3) g. (4) The most efficient technique for harvesting concentrated particles was the introduction of a frequency increment between two transducers to form a slowly sweeping pseudo-standing wave. The attractive inter-droplet ultrasonic standing wave force was employed to enhance the rate of aqueous biphasic cell separation and harvesting. The results help clarify the particle size, concentration, density and compressibility for which standing wave separation techniques can contribute either on a process engineering scale or on the scale of the manipulation of small particles for industrial and medical diagnostic procedures.

摘要

悬浮在声波驻波场中的细胞或颗粒会受到力的作用,这些力会将它们集中在相隔半个波长的位置。本研究的目的是:(1)优化条件并测试用于颗粒或细胞超声浓缩和分离的理论预测。(2)研究实验系统的放大。(3)确定悬浮液在不引起破坏有序的空化情况下可能承受的最大声压。(4)比较收集浓缩颗粒的技术效率。主要成果如下:(1)设计圆柱形容器内的声压分布,在驻波模式下使整个容器内的声压模式均匀重复,将悬浮的真核细胞或乳胶珠集中成块,位于宽容器的轴线上,并使所有颗粒团对声收集方式具有均匀响应。扩展了关于不同横向尺寸容器中颗粒行为(如移动到不同的优选位置)与比重和可压缩性的函数关系的理论,并通过实验得到证实。确定容器中的对流是控制1微米或更小尺寸颗粒时需要控制的一个变量。(2)从10毫升体积的模型进行放大研究得出结论,中等体积容器中的流动系统比放大的分批系统更有前景。(3)在1兆赫和3兆赫下,适用于悬浮液而不引起破坏有序的空化或过度传导流的最大声压会产生相当于约10³g离心场的力。(4)收集浓缩颗粒最有效的技术是在两个换能器之间引入频率增量,以形成缓慢扫过的伪驻波。利用有吸引力的液滴间超声驻波力提高双水相细胞分离和收集的速率。这些结果有助于阐明驻波分离技术在过程工程规模或工业和医学诊断程序中小颗粒操作规模上能够发挥作用的颗粒尺寸、浓度、密度和可压缩性。

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