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The topology of the contact seam of human erythrocytes adhered by dextran, an uncharged polymer, has been examined. Particular attention has been paid to the influence of electrostatic intermembrane interactions since their magnitude and range can be accurately estimated. Normal cells formed a continuous seam, whereas erythrocytes with pronase-modified glycocalices formed localized contact points on adhesion in 72 kDa dextran in buffered 145 mM NaCl. The dependence of the inter-contact distance lambda on dextran concentration [D] over the range 2-6% w/v, was given by lambda = C[D]-0.62, where C was a constant. The index of [D] was independent of dextran molecular mass over the range 20 to 450 kDa. The inter-contact distance for pronase-pretreated cells in 6% w/v 72 kDa dextran increased from 0.78 to 1.4 microns as [NaCl] was reduced through the range 145 to 90 mM and the suspending phase was maintained at isotonicity by using sorbitol to replace NaCl. The formation and lateral separation of the contact points are discussed from the perspective of linear interfacial instability theory. The theory allows a quantitative explanation for the experimentally observed dependence of inter-contact distance and of disturbance growth rate on change in electrostatic interaction. The results suggest that the dominant wavelength, determining the inter-contact distance, is established on approaching membranes when the layers of cell surface charge are separated by a perpendicular distance of < 14 nm (bilayer separation of 24 nm).

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Separation of particles from the suspending phase is of interest, among others, to clinical analysts. A system that enables manipulation of sub-micron sized particles in suspensions of analytical scale volume (10-50 microl) using a non-cavitating ultrasonic standing wave is described. Particle suspensions, contained in glass capillary tubes of 1-2 mm internal dimension, are treated on the axis of a tubular transducer generating a radial standing wave field at 4.5 MHz. Microparticles (of average diameter range 0.3-10 microm) suspended in buffer are concentrated within seconds at preferred regions separated by submillimetre distances. Concentration of suspended latex particles was inhibited in solutions containing protein at levels similar to those occurring in clinical specimens when the suspensions were sonicated in capillaries of circular cross-section. This effect was associated with acoustic streaming of the suspending fluid. Silica microparticles (more dense and less compressible than latex) could be concentrated in the presence of streaming. Latex particles concentrated readily in square cross-section capillaries where no streaming was observed. With sub-micron particles, the geometry of the sample chamber, the suspending phase composition and the size, density and compressibility of the microparticles all influence particle manipulation. The radial standing wave system has been used to enhance agglutination of antibody-coated latex microparticles in the presence of antigen allowing rapid and highly sensitive detection of clinically important biomolecules. The sensitivity of conventional diagnostic tests for microbial antigen has been improved by application of ultrasound and clinical utility has been demonstrated, in particular, for detection of meningitis-causing bacteria.

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