Cellular membranes have long served as an inspiration for nanomaterial research. The preparation of ultrathin polydopamine (PDA) films with integrated protein pores containing phospholipids and an embedded domain of a membrane protein glycophorin A as simplified cell membrane mimics is reported. Large area, ultrathin PDA films are obtained by electropolymerization on gold surfaces with 10-18 nm thickness and dimensions of up to 2.5 cm 2 . The films are transferred from gold to various other substrates such as nylon mesh, silicon, or substrates containing holes in the micrometer range, and they remain intact even after transfer. The novel transfer technique gives access to freestanding PDA films that remain stable even at the air interfaces with elastic moduli of ≈6-12 GPa, which are higher than any other PDA films reported before. As the PDA film thickness is within the range of cellular membranes, monodisperse protein nanopores, so-called "nanodiscs," are integrated as functional entities. These nanodisc-containing PDA films can serve as semipermeable films, in which the embedded pores control material transport. In the future, these simplified cell membrane mimics may offer structural investigations of the embedded membrane proteins to receive an improved understanding of protein-mediated transport processes in cellular membranes.

A photolithographic technique based on dry film photoresists for facile and low-cost patterning of microcantilever beams is presented. Dry film photoresists enable instantly homogenous photoresist coatings on flexible and 3D patterned substrate surfaces, represented here by microcantilever beams, which is otherwise challenging if conventional spin-coating of photoresist is utilized. Compared to alternative microtechnologies, such as focused ion beam milling or resist spray coating, our strategy is far less elaborate, fully compatible with routine additive and subtractive microfabrication processes and can be readily scaled. We show specifically microcantilever shape modification by CF 4 reactive ion etching, localized metal deposition in combination with conventional lift-off procedures as well as a utilization of patterned dry film photoresists as permanent microstructural elements. These microstructural elements are in particular flat-ended cylindrical dry resist micropillars created at the freestanding end of the cantilever beam that can be employed as scanning probes. The resist pillars enabled imaging of a 3T3 mouse fibroblast cell culture surface to determine their elastic force constants. Alongside UV-exposure by a conventional mask aligner, we also demonstrate dry film photoresist exposure by contact-free laser lithography eliminating possible substrate damage by photomask contact.

The actin cytoskeleton with its dynamic properties serves as the driving force for the movement and division of cells and gives the cell shape and structure. Disorders in the actin cytoskeleton occur in many diseases. Deeper understanding of its regulation is essential in order to better understand these biochemical processes. In our study, we use metal-induced energy transfer (MIET) as a tool to quantitatively examine the rarely considered third dimension of the actin cytoskeleton with nanometer accuracy. In particular, we investigate the influence of different drugs acting on the ROCK pathway on the three-dimensional actin organization. We find that cells treated with inhibitors have a lower actin height to the substrate while treatment with a stimulator for the ROCK pathway increases the actin height to the substrate, while the height of the membrane remains unchanged. This reveals the precise tuning of adhesion and cytoskeleton tension, which leads to a rich three-dimensional structural behaviour of the actin cytoskeleton. This finetuning is differentially affected by either inhibition or stimulation. The high axial resolution shows the importance of the precise finetuning of the actin cytoskeleton and the disturbed regulation of the ROCK pathway has a significant impact on the actin behavior in the z dimension.

Bordered pit membranes of angiosperm xylem are anisotropic, mesoporous media between neighbouring conduits, with a key role in long distance water transport. Yet, their mechanical properties are poorly understood. Here, we aim to quantify the stiffness of intervessel pit membranes over various growing seasons. By applying an AFM-based indentation technique “Quantitative Imaging” we measured the effective elastic modulus (Eeffective) of intervessel pit membranes of Clematis vitalba in dependence of size, age, and hydration state. The indentation-deformation behaviour was analysed with a non-linear membrane model, and paired with magnetic resonance imaging to visualise sap-filled and embolised vessels, while geometrical data of bordered pits were obtained using electron microscopy. Eeffective was transformed to the geometrically independent apparent elastic modulus Eapparent and to aspiration pressure Pb. The material stiffness (Eapparent) of fresh pit membranes was with 57 MPa considerably lower than previously suggested. The estimated pressure for pit membrane aspiration was 2.20+28 MPa. Pit membranes from older growth rings were shrunken, had a higher material stiffness and a lower aspiration pressure than current year ones, suggesting an irreversible, mechanical ageing process. This study provides an experimental-stiffness analysis of hydrated intervessel pit membranes in their native state. The estimated aspiration pressure suggests that membranes are not deflected under normal field conditions. Although absolute values should be interpreted carefully, our data suggest that pit membrane shrinkage implies increasing material stiffness, and highlight the dynamic changes of pit membrane mechanics and their complex, functional behaviour for fluid transport.

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