Axisymmetric Drop Shape Analysis (ADSA): An Outline.

Drop shape techniques for the measurement of interfacial tension are powerful, versatile and flexible. The shape of the drop/bubble depends on the balance between surface tension and external forces, e.g. gravity. This balance is reflected mathematically in the Laplace equation of capillarity. Axisymmetric Drop Shape Analysis (ADSA) is a commonly used drop shape technique. A streamlined version of the development of ADSA over the past several decades is presented to illustrate its validity and range of utility. Several configurations of interest will be considered and presented systematically. Shape and surface tension will be linked to a shape parameter based on proper concepts of differential geometry. The resulting shape parameter will be shown to allow determination of the range of applicability of such a drop shape method.

Phase Transitions in Dipalmitoylphosphatidylcholine Monolayers.

A self-assembled phospholipid monolayer at an air-water interface is a well-defined model system for studying surface thermodynamics, membrane biophysics, thin-film materials, and colloidal soft matter. Here we report a study of two-dimensional phase transitions in the dipalmitoylphosphatidylcholine (DPPC) monolayer at the air-water interface using a newly developed methodology called constrained drop surfactometry (CDS). CDS is superior to the classical Langmuir balance in its capacity for rigorous temperature control and leak-proof environments, thus making it an ideal alternative to the Langmuir balance for studying lipid polymorphism. In addition, we have developed a novel Langmuir-Blodgett (LB) transfer technique that allows the direct transfer of lipid monolayers from the droplet surface under well-controlled conditions. This LB transfer technique permits the direct visualization of phase coexistence in the DPPC monolayer. With these technological advances, we found that the two-dimensional phase behavior of the DPPC monolayer is analogous to the three-dimensional phase transition of a pure substance. This study has implications in the fundamental understanding of surface thermodynamics as well as applications such as self-assembled monolayers and pulmonary surfactant biophysics.

Laplacian drop shapes and effect of random perturbations on accuracy of surface tension measurement for different drop constellations.

Theoretical drop shapes are calculated for three drop constellations: pendant drops, constrained sessile drops, and unconstrained sessile drops. Based on total Gaussian curvature, shape parameter and critical shape parameter are discussed as a function of different drop sizes and surface tensions. The shape parameter is linked to physical parameters for every drop constellation. The as yet unavailable detailed dimensional analysis for the unconstrained sessile drop is presented. Results show that the unconstrained sessile drop shape depends on a dimensionless volume term and the contact angle. Random perturbations are introduced and the accuracy of surface tension measurement is assessed for precise and perturbed profiles of the three drop constellations. It is concluded that pendant drops are the best method for accurate surface tension measurement, followed by constrained sessile drops. The unconstrained sessile drops come last because they tend to be more spherical at low and moderate contact angles. Of course, unconstrained sessile drops are the only option if contact angles are to be measured.

A theory for the surface tensions and contact angles of hydrogen-bonding liquids.

The surface tensions of non-hydrogen-bonding, organic liquids can be accurately calculated from their electromagnetic properties, using an approximate form of the Lifshitz theory. A simple extension of this approach to the calculation of the surface tensions of hydrogen-bonding liquids is proposed. It is shown that the higher surface tensions of hydrogen-bonding liquids can be accounted for, with reasonable accuracy, by the increase in dispersion due to the shortened distance of approach between hydrogen-bonded atoms. Similar considerations allow calculations of contact angles on several low-energy solid surfaces in terms of molecular and electromagnetic properties. In accordance with well-known experimental observations, the calculated contact angles of both hydrogen-bonding and non-hydrogen-bonding liquids on the same low-energy surface nearly follow a single, smooth pattern.

Contact angle patterns on low-energy surfaces.

It is well-known that on a given low-energy solid surface, the contact angles of different organic liquids follow a regular pattern. The experimental evidence for this, and semi-empirical equations describing the pattern, are reviewed. Theoretical and computational efforts to explain the pattern are discussed, and a simplified analytical approach is presented. The main pattern of contact angles is seen to arise from two factors: a common combining rule for liquid-solid molecular interactions, and the reduced density of liquid molecules adjacent to a lower-energy solid surface. Irregular departures from the main pattern are due to chemical effects originating in molecular structure.

Total Gaussian curvature, drop shapes and the range of applicability of drop shape techniques.

Drop shape techniques are used extensively for surface tension measurement. It is well-documented that, as the drop/bubble shape becomes close to spherical, the performance of all drop shape techniques deteriorates. There have been efforts quantifying the range of applicability of drop techniques by studying the deviation of Laplacian drops from the spherical shape. A shape parameter was introduced in the literature and was modified several times to accommodate different drop constellations. However, new problems arise every time a new configuration is considered. Therefore, there is a need for a universal shape parameter applicable to pendant drops, sessile drops, liquid bridges as well as captive bubbles. In this work, the use of the total Gaussian curvature in a unified approach for the shape parameter is introduced for that purpose. The total Gaussian curvature is a dimensionless quantity that is commonly used in differential geometry and surface thermodynamics, and can be easily calculated for different Laplacian drop shapes. The new definition of the shape parameter using the total Gaussian curvature is applied here to both pendant and constrained sessile drops as an illustration. The analysis showed that the new definition is superior and reflects experimental results better than previous definitions, especially at extreme values of the Bond number.

Accuracy of surface tension measurement from drop shapes: the role of image analysis.

Axisymmetric Drop Shape Analysis (ADSA) has been extensively used for surface tension measurement. In essence, ADSA works by matching a theoretical profile of the drop to the extracted experimental profile, taking surface tension as an adjustable parameter. Of the three main building blocks of ADSA, i.e. edge detection, the numerical integration of the Laplace equation for generating theoretical curves and the optimization procedure, only edge detection (that extracts the drop profile line from the drop image) needs extensive study. For the purpose of this article, the numerical integration of the Laplace equation for generating theoretical curves and the optimization procedure will only require a minor effort. It is the aim of this paper to investigate how far the surface tension accuracy of drop shape techniques can be pushed by fine tuning and optimizing edge detection strategies for a given drop image. Two different aspects of edge detection are pursued here: sub-pixel resolution and pixel resolution. The effect of two sub-pixel resolution strategies, i.e. spline and sigmoid, on the accuracy of surface tension measurement is investigated. It is found that the number of pixel points in the fitting procedure of the sub-pixel resolution techniques is crucial, and its value should be determined based on the contrast of the image, i.e. the gray level difference between the drop and the background. On the pixel resolution side, two suitable and reliable edge detectors, i.e. Canny and SUSAN, are explored, and the effect of user-specified parameters of the edge detector on the accuracy of surface tension measurement is scrutinized. Based on the contrast of the image, an optimum value of the user-specified parameter of the edge detector, SUSAN, is suggested. Overall, an accuracy of 0.01mJ/m(2) is achievable for the surface tension determination by careful fine tuning of edge detection algorithms.

Contact angle hysteresis on randomly rough surfaces: a computational study.

Wetting is important in many applications, and the solid surfaces being wet invariably feature some amount of surface roughness. A free energy-based computational simulation is used to study the effect of roughness on wetting and especially contact angle hysteresis. On randomly rough, self-affine surfaces, it is found that hysteresis depends primarily on the value of the Wenzel roughness parameter r, increasing in proportion with r - 1. Micrometer-level roughness causes hysteresis of a few degrees.

Effect of surfactant concentration, compression ratio and compression rate on the surface activity and dynamic properties of a lung surfactant.

This paper reports dynamic surface tension experiments of a lung surfactant preparation, BLES, for a wide range of concentrations, compression ratios and compression rates. These experiments were performed using Axisymmetric Drop Shape Analysis-Constrained Sessile Drop (ADSA-CSD). The main purpose of the paper is to interpret the results in terms of physical parameters using the recently developed Compression-Relaxation Model (CRM). In the past, only the minimum surface tension was used generally for the characterization of lung surfactant films; however, this minimum value is not a physical parameter and depends on the compression protocol. CRM is based on the assumption that the dynamic surface tension response is governed by surface elasticities, adsorption and desorption of components of the lung surfactant. The ability of CRM to fit the surface tension response closely for a wide variety of parameters (compression ratio, compression rate and surfactant concentration) and produce sensible values for the elastic and kinetic parameters supports the validity of CRM.

Computation of contact lines on randomly heterogeneous surfaces.

Liquid wetting on real solid surfaces is significantly more complex than it is on the idealized surfaces of Young's equation. In the case of chemically heterogeneous solid surfaces, simulations of wetting have been carried out only for regularly patterned or one-dimensional surfaces. We describe a computational method for calculating advancing and receding contact lines on two-dimensional solid surfaces with arbitrary patterns of chemical heterogeneity. Results are verified against analytical solutions for homogeneous, single-defect, and striped surfaces. More practical surfaces with randomly placed high energy defects are also modeled. Realistic scatter in the contact angles as well as contact angle hysteresis and stick-slip motion are observed in the simulations. Hysteresis increases with the density of defects, but less so at high densities. The method allows prediction of wetting behavior from surface chemistry for a wide range of heterogeneous surfaces.

Range of validity of drop shape techniques for surface tension measurement.

Drop shape techniques are widely used for surface tension measurement. As the shape becomes close to spherical, the performance of drop shape techniques deteriorates. A quantitative criterion called shape parameter was previously introduced to quantify the meaning of "well-deformed" drops and "close to spherical" drops. In this paper, a modified definition of the shape parameter that does not depend on the radius of curvature at the drop apex is proposed. Such definition is applicable to different constellations of pendant and sessile drops/bubbles. Dimensional analysis is used to describe similarity in constrained sessile drop shapes and to express the problem using appropriate dimensionless groups. The proposed shape parameter is found to depend only on two dimensionless groups: the dimensionless volume (drop volume normalized by the cube of the holder radius) and the Bond number (using the drop holder's radius as the length scale). A critical shape parameter (minimum value of the shape parameter that guarantees a specified accuracy) is shown to depend only on Bond number. A set of experiments were performed with pure liquids to illustrate the change of the critical shape parameter with the Bond number.

Restoring the charge and surface activity of bovine lung extract surfactants with cationic and anionic polysaccharides.

Chitosan, a cationic polysaccharide, has been found to improve the surface activity of lung surfactant extracts in the presence of various inhibitors. It has been proposed that chitosan binds to anionic lipids (e.g. phosphatidyl glycerols) in lung surfactants, producing stable lipid films at the air-water interface. This binding also reverses the net charge of the surfactant aggregates, from negative to positive. Unfortunately, positively charged aggregates may adsorb or interact with the negatively charged epithelial tissue, leading to poor surfactant performance. To address this issue an anionic polysaccharide, dextran sulfate (dexS), was used as a secondary coating to reverse the charge of chitosan-lung surfactant extracts without affecting the surface activity of the preparation. The dynamic surface tension and zeta potential of bovine lipid extract surfactant (BLES) containing chitosan chloride (chiCl) and dexS were evaluated as a function of dexS concentration. These studies were conducted in the absence and presence of sodium bicarbonate buffer, and in the absence and presence of bovine serum used as model inhibitor. It was determined that using an appropriate concentration of dexS, especially at physiological pH, it is possible to restore the negative charge of the surfactant aggregates, and retain their surface activity, even in the presence of bovine serum. High concentrations of dexS affect the binding of chiCl to BLES, and the surface activity of the preparation.

Restoring the activity of serum-inhibited bovine lung extract surfactant (BLES) using cationic additives.

In this work four cationic additives were used to improve the surface activity of lung surfactants, particularly in the presence of bovine serum that was used as a model surfactant inhibitor. Two of those additives were chitosan in its soluble hydrochloride form with average molecular weights of 113kDa and 213kDa. The other two additives were cationic peptides, polylysine 50kDa and polymyxin B. These additives were added to bovine lipid extract surfactant (BLES) and the optimal additive-surfactant ratio was determined based on the minimum surface tension upon dynamic compression, carried out in a constrained sessile drop (CSD) device in the presence of 50 microl/ml serum. At the optimal ratio all the BLES-additive mixtures were able to achieve desirable minimum surface tensions. The optimal additive-surfactant ratios for the chitosan chlorides are consistent with a previously proposed patch model for the binding of the anionic lipids in BLES to the positive charges in chitosan. For the peptides, the optimal binding ratios were consistent with ratios established previously for the binding of these peptides to monolayers of anionic lipids. The optimal formulation containing these peptides were able to reach low minimum surface tension in systems containing 500 microl/ml of serum, matching the effectiveness of a lung surfactant extract that had not undergone post-separation processes and therefore contained all its proteins and lipids (complete lung surfactant).

A double injection ADSA-CSD methodology for lung surfactant inhibition and reversal studies.

This paper presents a continuation of the development of a drop shape method for film studies, ADSA-CSD (Axisymmetric Drop Shape Analysis-Constrained Sessile Drop). ADSA-CSD has certain advantages over conventional methods. The development presented here allows complete exchange of the subphase of a spread or adsorbed film. This feature allows certain studies relevant to lung surfactant research that cannot be readily performed by other means. The key feature of the design is a second capillary into the bulk of the drop to facilitate addition or removal of a secondary liquid. The development will be illustrated through studies concerning lung surfactant inhibition. After forming a sessile drop of a basic lung surfactant preparation, the bulk phase can be removed and exchanged for one containing different inhibitors. Such studies mimic the leakage of plasma and blood proteins into the alveolar spaces altering the surface activity of lung surfactant in a phenomenon called surfactant inhibition. The resistance of the lung surfactant to specific inhibitors can be readily evaluated using the method. The new method is also useful for surfactant reversal studies, i.e. the ability to restore the normal surface activity of an inhibited lung surfactant film by using special additives. Results show a distinctive difference between the inhibition when an inhibitor is mixed with and when it is injected under a preformed surfactant film. None of the inhibitors studied (serum, albumin, fibrinogen, and cholesterol) were able to penetrate a preexisting film formed by the basic preparation (BLES and protasan), while all of them can alter the surface activity of such preparation when mixed with the preparation. Preliminary results show that reversal of serum inhibition can be easily achieved and evaluated using the modified methodology.

Mixed DPPC/DPPG monolayers at very high film compression.

A drop shape technique using a constrained sessile drop constellation (ADSA-CSD) has been introduced as a superior technique for studying spread films specially at high collapse pressures [Saad et al. Langmuir 2008, 24, 10843-10850]. It has been shown that ADSA-CSD has certain advantages including the need only for small quantities of liquid and insoluble surfactants, the ability to measure very low surface tension values, easier deposition procedure, and leak-proof design. Here, this technique was applied to investigate mixed DPPC/DPPG monolayers to characterize the role of such molecules in maintaining stable film properties and surface activity of lung surfactant preparations. Results of compression isotherms were obtained for different DPPC/DPPG mixture ratios: 90/10, 80/20, 70/30, 60/40, and 50/50 in addition to pure DPPC and pure DPPG at room temperature of 24 degrees C. The ultimate collapse pressure of DPPC/DPPG mixtures was found to be 70.5 mJ/m2 (similar to pure DPPC) for the cases of low DPPG content (up to 20%). Increasing the DPPG content in the mixture (up to 40%) caused a slight decrease in the ultimate collapse pressure. However, further increase of DPPG in the mixture (50% or more) caused a sharp decrease in the ultimate collapse pressure to a value of 59.9 mJ/m2 (similar to pure DPPG). The change in film elasticity was also tracked for the range of mixture ratios studied. The physical reasons for such changes and the interaction between DPPC and DPPG molecules are discussed. The results also show a change in the film hysteresis upon successive compression and expansion cycles for different mixture ratios.

Tissue surface tension measurement by rigorous axisymmetric drop shape analysis.

Certain behaviours of embryonic cell aggregates can be modelled by ascribing to them a tissue surface tension, with each cell analogous to a liquid molecule. Under normal gravity, aggregates are nearly spherical, but they can be partially flattened in a centrifuge. This allows measurement of their tissue surface tensions by a drop shape method such as axisymmetric drop shape analysis (ADSA). We study ectodermal embryonic cells from the frog Xenopus laevis subjected to centrifugation at 100 x g and 200 x g. We show that ADSA can be applied to irregular aggregate profiles and compare results with those from a previous, simpler version called ADSA-IP. With a modification in the experimental method, the two algorithms give similar results and the aggregate profiles more closely follow Laplacian curves. The ADSA fitting error allows an estimate of the relative uncertainty in the results.

Axisymmetric drop shape analysis for estimating the surface tension of cell aggregates by centrifugation.

Biological tissues behave in certain respects like liquids. Consequently, the surface tension concept can be used to explain aspects of the in vitro and in vivo behavior of multicellular aggregates. Unfortunately, conventional methods of surface tension measurement cannot be readily applied to small cell aggregates. This difficulty can be overcome by an experimentally straightforward method consisting of centrifugation followed by axisymmetric drop shape analysis (ADSA). Since the aggregates typically show roughness, standard ADSA cannot be applied and we introduce a novel numerical method called ADSA-IP (ADSA for imperfect profile) for this purpose. To examine the new methodology, embryonic tissues from the gastrula of the frog, Xenopus laevis, deformed in the centrifuge are used. It is confirmed that surface tension measurements are independent of centrifugal force and aggregate size. Surface tension is measured for ectodermal cells in four sample batches, and varies between 1.1 and 7.7 mJ/m2. Surface tension is also measured for aggregates of cells expressing cytoplasmically truncated EP/C-cadherin, and is approximately half as large. In parallel, such aggregates show a reduction in convergent extension-driven elongation after activin treatment, reflecting diminished intercellular cohesion.

Axisymmetric drop shape analysis-constrained sessile drop (ADSA-CSD): a film balance technique for high collapse pressures.

Collapse pressure of insoluble monolayers is a property determined from surface pressure/area isotherms. Such isotherms are commonly measured by a Langmuir film balance or a drop shape technique using a pendant drop constellation (ADSA-PD). Here, a different embodiment of a drop shape analysis, called axisymmetric drop shape analysis-constrained sessile drop (ADSA-CSD) is used as a film balance. It is shown that ADSA-CSD has certain advantages over conventional methods. The ability to measure very low surface tension values (e.g., <2 mJ/m2), an easier deposition procedure than in a pendant drop setup, and leak-proof design make the constrained sessile drop constellation a better choice than the pendant drop constellation in many situations. Results of compression isotherms are obtained on three different monolayers: octadecanol, dipalmitoyl-phosphatidyl-choline (DPPC), and dipalmitoyl-phosphatidyl-glycerol (DPPG). The collapse pressures are found to be reproducible and in agreement with previous methods. For example, the collapse pressure of DPPC is found to be 70.2 mJ/m2. Such values are not achievable with a pendant drop. The collapse pressure of octadecanol is found to be 61.3 mJ/m2, while that of DPPG is 59.0 mJ/m2. The physical reasons for these differences are discussed. The results also show a distinctive difference between the onset of collapse and the ultimate collapse pressure (ultimate strength) of these films. ADSA-CSD allows detailed study of this collapse region.

Current perspectives in pulmonary surfactant--inhibition, enhancement and evaluation.

Pulmonary surfactant (PS) is a complicated mixture of approximately 90% lipids and 10% proteins. It plays an important role in maintaining normal respiratory mechanics by reducing alveolar surface tension to near-zero values. Supplementing exogenous surfactant to newborns suffering from respiratory distress syndrome (RDS), a leading cause of perinatal mortality, has completely altered neonatal care in industrialized countries. Surfactant therapy has also been applied to the acute respiratory distress syndrome (ARDS) but with only limited success. Biophysical studies suggest that surfactant inhibition is partially responsible for this unsatisfactory performance. This paper reviews the biophysical properties of functional and dysfunctional PS. The biophysical properties of PS are further limited to surface activity, i.e., properties related to highly dynamic and very low surface tensions. Three main perspectives are reviewed. (1) How does PS permit both rapid adsorption and the ability to reach very low surface tensions? (2) How is PS inactivated by different inhibitory substances and how can this inhibition be counteracted? A recent research focus of using water-soluble polymers as additives to enhance the surface activity of clinical PS and to overcome inhibition is extensively discussed. (3) Which in vivo, in situ, and in vitro methods are available for evaluating the surface activity of PS and what are their relative merits? A better understanding of the biophysical properties of functional and dysfunctional PS is important for the further development of surfactant therapy, especially for its potential application in ARDS.

Interaction between chitosan and bovine lung extract surfactants.

The interaction between a cationic polyelectrolyte, chitosan, and an exogenous bovine lung extract surfactant (BLES) was studied using dynamic compression/expansion cycles of dilute BLES preparations in a Constrained Sessile Drop (CSD) device equipped with an environmental chamber conditioned at 37 degrees C and 100% R.H. air. Under these conditions, dilute BLES preparations tend to produce variable and relatively high minimum surface tensions. Upon addition of "low" chitosan to BLES ratios, the minimum surface tension of BLES-chitosan preparations were consistently low (i.e. <5 mJ/m2), and the resulting surfactant monolayers (adsorbed at the air-water interface) were highly elastic and stable. However, the use of "high" chitosan to BLES ratios induced the collapse of the surfactant monolayer at high minimum surface tensions (i.e. >15 mJ/m2). The zeta potential of the lung surfactant aggregates in the subphase suggests that chitosan binds to the anionic lipids (phosphatidyl glycerols) in BLES, and that this binding is ultimately responsible for the changes in the surface activity (elasticity and stability) of these surfactant-polyelectrolyte mixtures. Furthermore the transition from "low" to "high" chitosan to BLES ratios correlates with the flocculation and de-flocculation of surfactant aggregates in the subphase. It is proposed that the aggregation/segregation of "patches" of anionic lipids in the surfactant monolayer produced at different chitosan to BLES ratios explains the enhancing/inhibitory effects of chitosan. These observations highlight the importance of electrostatic interactions in lung surfactant systems.