Locally Controlled Growth of Individual Lambda-Shaped Carbon Nanofibers.

The locally defined growth of carbon nanofibers with lambda shape in an open flame process is demonstrated. Via the growth time, the geometry of the structures can be tailored to a Λ- or λ-type shape. Microchannel cantilever spotting and dip-pen nanolithography are utilized for the deposition of catalytic salt NiCl2 · 6H2 O for locally controlled growth of lambda-shaped carbon nanofibers. Rigorous downscaling reveals a critical catalytic salt volume of 0.033 µm³, resulting in exactly one lambda-shaped carbon nanofiber at a highly predefined position. An empirical model explains the observed growth process.

Site-Specific Surface Functionalization via Microchannel Cantilever Spotting (µCS): Comparison between Azide-Alkyne and Thiol-Alkyne Click Chemistry Reactions.

Different types of click chemistry reactions are proposed and used for the functionalization of surfaces and materials, and covalent attachment of organic molecules. In the present work, two different catalyst-free click approaches, namely azide-alkyne and thiol-alkyne click chemistry are studied and compared for the immobilization of microarrays of azide or thiol inks on functionalized glass surfaces. For this purpose, the surface of glass is first functionalized with dibenzocyclooctyne-acid (DBCO-acid), a cyclooctyne with a carboxyl group. Then, the DBCO-terminated surfaces are functionalized via microchannel cantilever spotting with different fluorescent and nonfluorescent azide and thiol inks. Although both routes work reliably for surface functionalization, the protein binding experiments reveal that using a thiol-alkyne route will obtain the highest surface density of molecular immobilization in such spotting approaches. The obtained achievements and results from this work can be used for design and manufacturing of microscale patterns suitable for biomedical and biological applications.

Polymer Pen Lithography with Lipids for Large-Area Gradient Patterns.

Gradient patterns comprising bioactive compounds over comparably (in regard to a cell size) large areas are key for many applications in the biomedical sector, in particular, for cell screening assays, guidance, and migration experiments. Polymer pen lithography (PPL) as an inherent highly parallel and large area technique has a great potential to serve in the fabrication of such patterns. We present strategies for the printing of functional phospholipid patterns via PPL that provide tunable feature size and feature density gradients over surface areas of several square millimeters. By controlling the printing parameters, two transfer modes can be achieved. Each of these modes leads to different feature morphologies. By increasing the force applied to the elastomeric pens, which increases the tip-surface contact area and boosts the ink delivery rate, a switch between a dip-pen nanolithography (DPN) and a microcontact printing (µCP) transfer mode can be induced. A careful inking procedure ensuring a homogeneous and not-too-high ink-load on the PPL stamp ensures a membrane-spreading dominated transfer mode, which, used in combination with smooth and hydrophilic substrates, generates features with constant height, independently of the applied force of the pens. Ultimately, this allows us to obtain a gradient of feature sizes over a mm2 substrate, all having the same height on the order of that of a biological cellular membrane. These strategies allow the construction of membrane structures by direct transfer of the lipid mixture to the substrate, without requiring previous substrate functionalization, in contrast to other molecular inks, where structure is directly determined by the printing process itself. The patterns are demonstrated to be viable for subsequent protein binding, therefore adding to a flexible feature library when gradients of protein presentation are desired.

Large-scale parallel surface functionalization of goblet-type whispering gallery mode microcavity arrays for biosensing applications.

A novel surface functionalization technique is presented for large-scale selective molecule deposition onto whispering gallery mode microgoblet cavities. The parallel technique allows damage-free individual functionalization of the cavities, arranged on-chip in densely packaged arrays. As the stamp pad a glass slide is utilized, bearing phospholipids with different functional head groups. Coated microcavities are characterized and demonstrated as biosensors.

Design of plasmonic grating structures towards optimum signal discrimination for biosensing applications.

Sensors based on surface plasmon resonances (SPRs) have proven themselves as promising devices for molecular investigations - still there is potential to determine the geometrical parameter set for optimal sensing performance. Here we propose a comprehensive design rule for one-dimensional plasmonic grating structures. We present an analytical approach, which allows for estimation of the grating parameters for best SPR coupling efficiency for any geometry and design wavelength. On the example of sinusoidal gratings, we expand this solution and discuss numerically and experimentally, how the grating modulation depth can be refined to achieve optimal signal resolution. Finally, we propose a benchmark factor to assess the sensor performance, which can be applied to any sensing scheme utilizing resonances, allowing for comparison of different technological platforms.

Low-cost label-free biosensors using photonic crystals embedded between crossed polarizers.

There is a strong need for low-cost biosensors to enable rapid, on-site analysis of biological, biomedical, or chemical substances. We propose a platform for label-free optical biosensors based on applying the analyte onto a surface-functionalized photonic crystal slab and performing a transmission measurement with two crossed polarization filters. This dark-field approach allows for efficient background suppression as only the photonic crystal guided-mode resonances interacting with the functionalized surface experience significant polarization rotation. We present a compact biosensor demonstrator using a low-cost light emitting diode and a simple photodiode capable of detecting the binding kinetics of a 2.5 nM solution of the protein streptavidin on a biotin-functionalized photonic crystal surface.

Writing Behavior of Phospholipids in Polymer Pen Lithography (PPL) for Bioactive Micropatterns.

Lipid-based membranes play crucial roles in regulating the interface between cells and their external environment, the communication within cells, and cellular sensing. To study these important processes, various lipid-based artificial membrane models have been developed in recent years and, indeed, large-area arrays of supported lipid bilayers suit the needs of many of these studies remarkably well. Here, the direct-write scanning probe lithography technique called polymer pen lithography (PPL) was used as a tool for the creation of lipid micropatterns over large areas via polymer-stamp-mediated transfer of lipid-containing inks onto glass substrates. In order to better understand and control the lipid transfer in PPL, we conducted a systematic study of the influence of dwell time (i.e., duration of contact between tip and sample), humidity, and printing pressure on the outcome of PPL with phospholipids and discuss results in comparison to the more often studied dip-pen nanolithography with phospholipids. This is the first systematic study in phospholipid printing with PPL. Biocompatibility of the obtained substrates with up to two different ink compositions was demonstrated. The patterns are suitable to serve as a platform for mast cell activation experiments.

Reconfigurable microfluidic photonic crystal slab cavities.

We demonstrate the spectral and spatial reconfigurability of photonic crystal double-heterostructure cavities in silicon by microfluidic infiltration of selected air holes. The lengths of the microfluidic cavities are changed by adjusting the region of infiltrated holes in steps of several microns. We systematically investigate the spectral signature of these cavities, showing high Q-factor resonances for a broad range of cavity lengths. The fluid can be removed by immersing the device in toluene, offering complete reconfigurability. Our cavity writing technique allows for tolerances in the infiltration process and provides flexibility as it can be employed at any time after photonic crystal fabrication.

Clickable Antifouling Polymer Brushes for Polymer Pen Lithography.

Protein-repellent reactive surfaces that promote localized specific binding are highly desirable for applications in the biomedical field. Nonspecific adhesion will compromise the function of bioactive surfaces, leading to ambiguous results of binding assays and negating the binding specificity of patterned cell-adhesive motives. Localized specific binding is often achieved by attaching a linker to the surface, and the other side of the linker is used to bind specifically to a desired functional agent, as e.g. proteins, antibodies, and fluorophores, depending on the function required by the application. We present a protein-repellent polymer brush enabling highly specific covalent surface immobilization of biorecognition elements by strain-promoted alkyne-azide cycloaddition click chemistry for selective protein adhesion. The protein-repellent polymer brush is functionalized by highly localized molecular binding sites in the low micrometer range using polymer pen lithography (PPL). Because of the massive parallelization of writing pens, the tunable PPL printed patterns can span over square centimeter areas. The selective binding of the protein streptavidin to these surface sites is demonstrated while the remaining polymer brush surface is resisting nonspecific adsorption without any prior blocking by bovine serum albumin (BSA). In contrast to the widely used BSA blocking, the reactive polymer brushes are able to significantly reduce nonspecific protein adsorption, which is the cause of biofouling. This was achieved for solutions of single proteins as well as complex biological fluids. The remarkable fouling resistance of the polymer brushes has the potential to improve the multiplexing capabilities of protein probes and therefore impact biomedical research and applications.

Densely Packed Microgoblet Laser Pairs for Cross-Referenced Biomolecular Detection.

Microgoblet laser pairs are presented for cross-referenced on-chip biomolecular sensing. Parallel readout of the micro-lasers facilitates effective mutual filtering of highly localized refractive index and temperature fluctuations in the analyte. Cross-referenced detection of two different types of proteins and complete chemical transducer reconfiguration is demonstrated. Selective surface functionalization of the individual lasers with high spatial accuracy is achieved by aligned microcontact stamping.

On-chip microlasers for biomolecular detection via highly localized deposition of a multifunctional phospholipid ink.

We report on a novel approach to realize on-chip microlasers, by applying highly localized and material-saving surface functionalization of passive photonic whispering gallery mode microresonators. We apply dip-pen nanolithography on a true three-dimensional structure. We coat solely the light-guiding circumference of pre-fabricated poly(methyl methacrylate) resonators with a multifunctional molecular ink. The functionalization is performed in one single fabrication step and simultaneously provides optical gain as well as molecular binding selectivity. This allows for a direct and flexible realization of on-chip microlasers, which can be utilized as biosensors in optofluidic lab-on-a-chip applications. In a proof-of-concept we show how this highly localized molecule deposition suffices for low-threshold lasing in air and water, and demonstrate the capability of the ink-lasers as biosensors in a biotin-streptavidin binding experiment.

High-Q microfluidic cavities in silicon-based two-dimensional photonic crystal structures.

We demonstrate postprocessed microfluidic double-heterostructure cavities in silicon-based photonic crystal slab waveguides. The cavity structure is realized by selective fluid infiltration of air holes using a glass microtip, resulting in a local change of the average refractive index of the photonic crystal. The microcavities are probed by evanescent coupling from a silica nanowire. An intrinsic quality factor of 57,000 has been derived from our measurements, representing what we believe to be the largest value observed in microfluidic photonic crystal cavities to date.