Massively parallel cantilever-free atomic force microscopy.

Resolution and field-of-view often represent a fundamental tradeoff in microscopy. Atomic force microscopy (AFM), in which a cantilevered probe deflects under the influence of local forces as it scans across a substrate, is a key example of this tradeoff with high resolution imaging being largely limited to small areas. Despite the tremendous impact of AFM in fields including materials science, biology, and surface science, the limitation in imaging area has remained a key barrier to studying samples with intricate hierarchical structure. Here, we show that massively parallel AFM with >1000 probes is possible through the combination of a cantilever-free probe architecture and a scalable optical method for detecting probe-sample contact. Specifically, optically reflective conical probes on a comparatively compliant film are found to comprise a distributed optical lever that translates probe motion into an optical signal that provides sub-10 nm vertical precision. The scalability of this approach makes it well suited for imaging applications that require high resolution over large areas.

Stiffness of HIV-1 Mimicking Polymer Nanoparticles Modulates Ganglioside-Mediated Cellular Uptake and Trafficking.

The monosialodihexosylganglioside, GM3, and its binding to CD169 (Siglec-1) have been indicated as key factors in the glycoprotein-independent sequestration of the human immunodeficiency virus-1 (HIV-1) in virus-containing compartments (VCCs) in myeloid cells. Here, lipid-wrapped polymer nanoparticles (NPs) are applied as a virus-mimicking model to characterize the effect of core stiffness on NP uptake and intracellular fate triggered by GM3-CD169 binding in macrophages. GM3-functionalized lipid-wrapped NPs are assembled with poly(lactic-co-glycolic) acid (PLGA) as well as with low and high molecular weight polylactic acid (PLAlMW and PLAhMW) cores. The NPs have an average diameter of 146 ± 17 nm and comparable surface properties defined by the self-assembled lipid layer. Due to differences in the glass transition temperature, the Young's modulus (E) differs substantially under physiological conditions between PLGA (E PLGA = 60 ± 32 MPa), PLAlMW (E PLA lMW = 86 ± 25 MPa), and PLAhMW (E PLA hMW = 1.41 ± 0.67 GPa) NPs. Only the stiff GM3-presenting PLAhMW NPs but not the softer PLGA or PLAlMW NPs avoid a lysosomal pathway and localize in tetraspanin (CD9)-positive compartments that resemble VCCs. These observations suggest that GM3-CD169-induced sequestration of NPs in nonlysosomal compartments is not entirely determined by ligand-receptor interactions but also depends on core stiffness.

Physiologically Relevant Mechanics of Biodegradable Polyester Nanoparticles.

Despite the extensive use of biodegradable polyester nanoparticles for drug delivery, and reports of the strong influence of nanoparticle mechanics on nano-bio interactions, there is a lack of systematic studies on the mechanics of these nanoparticles under physiologically relevant conditions. Here, we report indentation experiments on poly(lactic acid) and poly(lactide-co-glycolide) nanoparticles using atomic force microscopy. While dried nanoparticles were found to be rigid at room temperature, their elastic modulus was found to decrease by as much as 30 fold under simulated physiological conditions (i.e., in water at 37 °C). Differential scanning calorimetry confirms that this softening can be attributed to the glass transition of the nanoparticles. Using a combination of mechanical and thermoanalytical characterization, the plasticizing effects of miniaturization, molecular weight, and immersion in water were investigated. Collectively, these experiments provide insight for experimentalists exploring the relationship between polymer nanoparticle mechanics and in vivo behavior.

High-Throughput Multiobjective Optimization of Patterned Multifunctional Surfaces.

The realization and optimization of multifunctional materials is difficult, especially when the functionalities are directly incompatible. For example, it is challenging to make surfaces both enzymatically active and water repellent, as these two properties are directly competitive because of the hydrophilic nature of the enzyme-laden surfaces. Patterning discrete domains of distinct functionalities can represent a path to multifunctionality, but the innumerable possible domain permutations present a major barrier to optimizing performance. Here, we develop a high-throughput approach for exploring patterned multifunctional surfaces that is inspired by the microtiter plate architecture. As a model system, patterned surfaces are realized with horseradish peroxidase-decorated domains amidst a background of hydrophobic fluorinated self-assembled monolayers. In experiments exploring effects of pattern geometry, the measured enzyme activity is dependent only on the surface coverage. In contrast, roll-off behavior strongly depends on the parameters of the pattern geometry. Importantly, this finding enables the precise tailoring of distinct wetting behavior of the surfaces in a manner that is independent of their enzymatic activity. The high-throughput nature of the platform facilitates multiobjective optimization of surface functionalities in a general and flexible manner.

Confinement-Induced Stiffening of Elastomer Thin Films.

While entropy is understood to dictate the elastic mechanics of elastomers, a consensus has not been reached regarding why they stiffen when confined to thin films or even the magnitude of this effect. Here, we present a combined computational and experimental approach to measure the confinement-induced stiffening of thin elastomer films using nanoindentation. First, we use finite element computations to modify the adhesion-free contact model to include the presence of a rigid substrate and verify this correction using indentation experiments on macroscopic polydimethylsiloxane (PDMS) films. Next, we validate this correction factor in indentation experiments where adhesion plays a major role through a series of experiments on micrometer-scale PDMS films. With this correction factor in hand, we perform an extensive series of nanoindentation experiments on submicrometer thickness films of varying cross-link density using a variety of probe radii and probe surface energies. We observe a consistent stiffening of all PDMS thin films that, interestingly, converges to a consistent elastic modulus at thicknesses ∼100 nm regardless of the cross-link density. To explain these results, we propose a surface cross-linking model where the density of crosslinks between chains increases near the surface of the film because of oxidation-induced bonds between chains. Importantly, the apparent moduli predicted by this entropic model are in good agreement with all experimental results. Collectively, these results shed new light on the nanomechanics of elastomers and provide a general method for predicting the size-dependent effective moduli of soft polymers.

Design and Realization of 3D Printed AFM Probes.

Atomic force microscope (AFM) probes and AFM imaging by extension are the product of exceptionally refined silicon micromachining, but are also restricted by the limitations of these fabrication techniques. Here, the nanoscale additive manufacturing technique direct laser writing is explored as a method to print monolithic cantilevered probes for AFM. Not only are 3D printed probes found to function effectively for AFM, but they also confer several advantages, most notably the ability to image in intermittent contact mode with a bandwidth approximately ten times larger than analogous silicon probes. In addition, the arbitrary structural control afforded by 3D printing is found to enable programming the modal structure of the probe, a capability that can be useful in the context of resonantly amplifying nonlinear tip-sample interactions. Collectively, these results show that 3D printed probes complement those produced using conventional silicon micromachining and open the door to new imaging techniques.

Photoactuated Pens for Molecular Printing.

The photoactuation of pen arrays made of polydimethylsiloxane carbon nanotube composites is explored, and the first demonstration of photoactuated pens for molecular printing is reported. Photoactuation of these composites is characterized using atomic force microscopy and found to produce microscale motion in response to modest illumination, with an actuation efficiency as high as 200 nm mW-1 on the sub-1 s time scale. Arrays of composite pens are synthesized and it is found that local illumination is capable of moving selected pens by more than 3 µm out of the plane, bringing them into contact to perform controllable and high quality printing while completely shutting off the nonilluminated counterparts. In light of the scalability limitations of nanolithography, this work presents an important step and paves the way for arbitrary control of individual pens in massive arrays. As an example of a scalable soft actuator, this approach can also aid progress in other fields such as soft robotics and microfluidics.

Elasticity and failure of liquid marbles: influence of particle coating and marble volume.

When coated with microscale hydrophobic particles, macroscopic liquid droplets can become non-wetting liquid marbles that exhibit an array of fascinating solid-like properties. Specifically, the force required to uniaxially compress liquid marbles depends on their volume, but it is unclear if the particle coating plays a role. In contrast, the failure of marbles upon compression does depend on the particle coating, but the conditions for failure do not appear to change with marble volume. Here, we experimentally study the elastic deformation and failure of liquid marbles and, by applying a doubly truncated oblate spheroid model to quantify their surface area, explore the role of marble volume and particle composition. First, we find that the work required to compress liquid marbles agrees with the product of the core fluid surface tension and the change in the marble surface area, validating that the elastic mechanics of liquid marbles is independent of the particle coating. Next, we study marble failure by measuring their ductility as quantified by the maximum fractional increase in marble surface area prior to rupture. Not only does marble ductility depend on the particle coating, but it also depends on marble volume with smaller marbles being more ductile. This size effect is attributed to an interaction between marble curvature and particle rafts held together by interparticle forces. These results illuminate new avenues to tailor the rupture of liquid marbles for applications spanning smart fluid handling and pollution mitigation.