Acoustic tethering of microorganisms.

We show how to construct and apply a setup to acoustically tether and enable behavioral observations of individual microorganisms using simple laboratory equipment and a standard light microscope. We explore the capability of the setup with the freely swimming dinoflagellate Alexandrium minutum as the study organism. The setup allows us to tether cells in focus in the mid-plane of the sample chamber and make observations of individual organisms at high magnification without affecting their flagellar beat frequencies. We discuss the prospect of the method to explore appendage motion and swimming kinematics of other flagellates and ciliates, and we argue that the method will be applicable to a broad range of cell sizes and shapes.

Two-dimensional order in mammalian pre-ocular tear film.

We report a grazing incidence x-ray diffraction (GIXD) investigation of the surface lipid layer of the pre-ocular tear film. For the first time we demonstrate the existence of 2D order over a wide range of surface pressures in this system, with typical spicing of 3.75A and 4.16A independent of the monolayer surface pressure. Analogous lipid ordering is also found in an artificial lipid mixture of the major lipid components of the tear film, suggesting that the 2D ordering is set by generic lipid-lipid interactions. Fluorescence microscopy of the natural and artificial tear film mixture reveals the co-existence of a dilute and a much more condensed phase in the amphiphilic lipid matrix over the pressure range of 15-45mN/m investigated by GIXD, plus an additional structure due to the much more hydrophobic part of the mixture. This evidence supports the previous hypothesis that tear film has a layered structure.

Viscoelastic properties of insoluble amphiphiles at the air/water interface.

The effects of the presence of a molecular monolayer on the dilatational properties of the air/water interface have been investigated. Two water insoluble amphiphiles, dipalmitoyl phosphatidyl choline and quercetin 3-O-palmitate, were spread onto a pendant drop and the dynamic surface pressure was measured by means of drop shape analysis. The surface dilatational elasticity and viscosity of the spread monolayers were also determined by the oscillating drop technique. Constraints on the range of measuring conditions were investigated and we demonstrated that the pressure-area isotherms derived from oscillatory dynamic measurements display phase behaviour similar to that found in equilibrium measurements, albeit at reduced resolution. Both the amphiphiles formed purely elastic films that were characterised by a dilatational modulus that depended on the surface concentration and obeyed a power scaling law. The exponent of the relationship could be related to the thermodynamic conditions prevailing at the interface. The phospholipid monolayer scaling exponent was 2.8 in a temperature range of 20-26 degrees C indicates a favourable solvency of molecules in the bidimensional matrix. A very high scaling exponent (11.8 at 7 degrees C) for quercetin palmitate was interpreted assuming that molecules self-organise in fibre-like structures. This interface structure and the phase behaviour was found consistent with observations of the surface film obtained by Brewster angle microscopy. The structured quercetin 3-O-palmitate monolayers are disrupted by temperature increase or by adding a 0.2 molar fraction of the immiscible dipalmitoyl phosphatidyl choline.

Insertion of tear proteins into a meibomian lipids film.

The eyelid meibomian gland secretions form the outer layer of the tear film. That layer functions as a lubricant during a blink, and as a barrier against intrusion of foreign bodies. The lipid film is also exposed to proteins present in the aqueous phase that may adsorb there, and thus form an integral part of the surface of the tear film, or possibly, cause disruption to the outermost layer. Therefore, the adsorption of tear proteins to the meibomian lipid layer was object of the present investigation. A model tear was set up coating a pendant drop of saline with a film of meibomian lipids and measuring variations of the interfacial pressure after the injection of tear proteins into the aqueous subphase at their physiological concentration. All tear proteins adsorbed at the interface causing the initial surface pressure to increase. For each protein, a limiting surface pressure at which a given protein was no longer able to insert into the lipid layer was found. Among the proteins tested, lipocalin was the most surface active one and inserted into the lipid layer in the whole range of surface pressure exerted by the meibomian lipid mixture. Lactoferrin, lysozyme and IgA also interacted with the lipids whereas albumin interacted more weakly. The timescale of the protein insertion into the lipid layer was of the order of 10(2) s. It was hypothesized that protein adsorption at the interface could be associated with structural changes. Indeed, the enzymatic activity of lysozyme was maintained in the presence of an outermost meibomian lipid layer that prevented its denaturation while exposure at the air/aqueous interface induced significant lysozime degradation. meibomian lipid composition is therefore functional to maintain tear proteins activity.