Versatile Wettability Gradients Prepared by Chemical Modification of Polymer Brushes on Polymer Foils

A method to create a wettability gradient by variation of the chemical functionality in a polymer brush is presented. A poly(N-methyl-vinylpyridinium) (QP4VP) brush was created on a poly(ethylene-alt-tetrafluoroethylene) (ETFE) foil by the grafting of 4-vinylpyridine and subsequent quaternization. The instability of QP4VP, a strong polyelectrolyte, in alkaline media was exploited to transform it to the neutral poly(vinyl(N-methyl-2-pyridone)) (PVMP), as confirmed with ATR-IR spectroscopy. The slow transformation resulted in a substantial, time-dependent decrease in wettability. A nearly linear gradient in water contact angle (CA) was created by immersion of a QP4VP brush modified sample into a sodium hydroxide solution, resulting in CAs ranging from 10° to 60°. The concurrent decrease in the number of charged functional groups along the gradient was characterized by loading an anionic dye into the polymer brush and measuring the UV transmittance of the sample. The versatility of the wettability gradient was demonstrated by exchanging the counterions of the N-methyl-vinylpyridinium groups, whereby a reversal of gradient direction was reproducibly achieved

Light-Responsive Polymer Surfaces via Postpolymerization Modification of Grafted Polymer-Brush Structures

Light-induced, spatially well-defined, reversible switching of surface properties enables the creation of remote-controlled smart surfaces. We have taken advantage of the unique high-resolution structuring capabilities of extreme ultraviolet (EUV) interference lithography to produce nanostructured photoresponsive polymer brushes. Patterns of poly­(glycidyl methacrylate) (PGMA) and poly­(methacrylic acid) (PMAA) were grafted from two different 100 μm thick fluoropolymer substrates by means of a radiation-initiated, grafting-from approach based on free-radical polymerization (FRP). Photochromic properties were introduced via novel one- or two-step postpolymerization modifications with spiropyran (SP) derivatives, which allowed us to control the number of photochromic groups on the polymer brushes. Depending on the degree of functionalization and the local chemical environment, the SP moieties can open upon UV-light exposure to form zwitterionic, deeply colored, and fluorescent merocyanines (MCs) and reclose to the colorless SP configuration via thermal or visible light-induced relaxation. Switching kinetics were studied by means of time-resolved fluorescence microscopy and compared with kinetic measurements of the SP moiety in solution. The results indicated the importance, for the intensity of the switching, of the local chemical environment provided by both the polymer brush and added solvents, and showed the predominant influence on the ring-closing kinetics of polar solvents, which stabilize the MC form. To allow further characterization of the polymer-brush arrangements on a macroscopic scale, similar, but unstructured brush systems were grafted from fluoropolymers after large-area activation using EUV radiation or argon plasma. All steps of the postpolymerization modification were characterized in detail using attenuated total reflection infrared (ATR-IR) spectroscopy. Furthermore, a light-induced reversible static-contact-angle switch with a range of up to 15° for PGMA-SP brushes and up to 30° for PMA-SP brushes was demonstrated upon alternating UV- and visible-light irradiation

From pH- to Light-Response: Postpolymerization Modification of Polymer Brushes Grafted onto Microporous Polymeric Membranes

A microporous pH- and light-responsive membrane that enables remote control over its interfacial properties has been fabricated. pH-Responsiveness was imparted to a porous polypropylene film via grafting of poly­(methacrylic acid) brushes from the substrate using argon-plasma-induced free-radical graft polymerization. Morphological changes as a function of grafting level were analyzed using atomic force microscopy. Conversion into a light-responsive membrane was performed via postpolymerization modification to covalently attach photochromic spiropyran moieties to the grafted polymer brushes. Reversible switches in wettability and permeability were determined upon changing from acidic to basic pH or upon alternating UV- and visible-light irradiation. Additionally, light-responsive membranes show a switch in color upon UV exposure

From pH- to Light-Response: Postpolymerization Modification of Polymer Brushes Grafted onto Microporous Polymeric Membranes

A microporous pH- and light-responsive membrane that enables remote control over its interfacial properties has been fabricated. pH-Responsiveness was imparted to a porous polypropylene film via grafting of poly­(methacrylic acid) brushes from the substrate using argon-plasma-induced free-radical graft polymerization. Morphological changes as a function of grafting level were analyzed using atomic force microscopy. Conversion into a light-responsive membrane was performed via postpolymerization modification to covalently attach photochromic spiropyran moieties to the grafted polymer brushes. Reversible switches in wettability and permeability were determined upon changing from acidic to basic pH or upon alternating UV- and visible-light irradiation. Additionally, light-responsive membranes show a switch in color upon UV exposure

Preparation of Micro- and Nanopatterns of Polymer Chains Grafted onto Flexible Polymer Substrates

In this work a simple novel method for preparing micro- and nanoscale patterns of polymer chains grafted onto flexible polymer substrates is described. A combination of the two techniques of radiation grafting and “grafting-from” has been made. This combination makes it possible to prepare grafted structures having micro- or nanoscale lateral dimensions that are determined by the electron beam or X-ray irradiation patterns used. The height of the grafted features can be controlled by the irradiation dose or such grafting reaction conditions as time, temperature, or monomer concentration. Our first results for nanopatterned samples demonstrate resolution comparable to those of other polymer-based lithography processes

Poly(methyl methacrylate)-Based Nanofluidic Device for Rapid and Multiplexed Serological Antibody Detection of SARS-CoV‑2

The outbreak of SARS-CoV-2 has emphasized the value of point-of-care diagnostics, as well as reliable and cost-effective serological antibody tests, to monitor the viral spread and contain pandemics and endemics. Here, we present a three-dimensional (3D) nanofluidic device for rapid and multiplexed detection of viral antibodies. The device is made from poly­(methyl methacrylate) and contains 3D fluidic channels with nanoscale topography variations on the millimeter length scale, enabled by combining gray-scale e-beam lithography and nanoimprint lithography. It works with capillary pumps only and does not require a complex microfluidic setup and pumps, which hinder the widespread usage of micro- and nanofluidic devices. The device is designed to size dependently immobilize particles from a multiparticle mixture at predefined positions in nanochannels, resulting in distinct trapping lines. We show that it can be used as an on-chip fluorescence-linked immunosorbent assay for highly specific and sensitive multiplexed detection of serological antibodies against different viral proteins. Further test flexibility is demonstrated by on-bead color multiplexing for simultaneous detection of IgG and IgM antibodies in convalescent human serum. The particle sorting is further leveraged to enable concurrent detection of anti-spike (SARS-CoV-2) and anti-hemagglutinin (influenza A) antibodies. The device’s applications can be further extended to detect a large variety of diseases simultaneously in a reliable and straightforward manner

Poly(methyl methacrylate)-Based Nanofluidic Device for Rapid and Multiplexed Serological Antibody Detection of SARS-CoV‑2

The outbreak of SARS-CoV-2 has emphasized the value of point-of-care diagnostics, as well as reliable and cost-effective serological antibody tests, to monitor the viral spread and contain pandemics and endemics. Here, we present a three-dimensional (3D) nanofluidic device for rapid and multiplexed detection of viral antibodies. The device is made from poly­(methyl methacrylate) and contains 3D fluidic channels with nanoscale topography variations on the millimeter length scale, enabled by combining gray-scale e-beam lithography and nanoimprint lithography. It works with capillary pumps only and does not require a complex microfluidic setup and pumps, which hinder the widespread usage of micro- and nanofluidic devices. The device is designed to size dependently immobilize particles from a multiparticle mixture at predefined positions in nanochannels, resulting in distinct trapping lines. We show that it can be used as an on-chip fluorescence-linked immunosorbent assay for highly specific and sensitive multiplexed detection of serological antibodies against different viral proteins. Further test flexibility is demonstrated by on-bead color multiplexing for simultaneous detection of IgG and IgM antibodies in convalescent human serum. The particle sorting is further leveraged to enable concurrent detection of anti-spike (SARS-CoV-2) and anti-hemagglutinin (influenza A) antibodies. The device’s applications can be further extended to detect a large variety of diseases simultaneously in a reliable and straightforward manner

Fabrication of Thiol–Ene “Clickable” Copolymer-Brush Nanostructures on Polymeric Substrates via Extreme Ultraviolet Interference Lithography

We demonstrate a new approach to grafting thiol-reactive nanopatterned copolymer-brush structures on polymeric substrates by means of extreme ultraviolet (EUV) interference lithography. The copolymer brushes were designed to contain maleimide functional groups as thiol-reactive centers. Fluoropolymer films were exposed to EUV radiation at the X-ray interference lithography beamline (XIL-II) at the Swiss Light Source, in order to create radical patterns on their surfaces. The radicals served as initiators for the copolymerization of thiol–ene “clickable” brushes, composed of a furan-protected maleimide monomer (FuMaMA) and different methacrylates, namely, methyl methacrylate (MMA), ethylene glycol methyl ether methacrylate (EGMA), or poly­(ethylene glycol) methyl ether methacrylate (PEGMA). Copolymerization with ethylene-glycol-containing monomers provides antibiofouling properties to these surfaces. The number of reactive centers on the grafted brush structures can be tailored by varying the monomer ratios in the feed. Grafted copolymers were characterized by using attenuated total reflection infrared (ATR-IR) spectroscopy. The reactive maleimide methacrylate (MaMA) units were utilized to conjugate thiol-containing moieties using the nucleophilic Michael-addition reaction, which proceeds at room temperature without the need for any metal-based catalyst. Using this approach, a variety of functionalities was introduced to yield polyelectrolytes, as well as fluorescent and light-responsive polymer-brush structures. Functionalization of the brush structures was demonstrated via ATR-IR and UV–vis spectroscopy and fluorescence microscopy, and was also indicated by a color switch. Furthermore, grafted surfaces were generated via plasma activation, showing a strongly increased wettability for polyelectrolytes and a reversible switch in static water contact angle (CA) of up to 18° for P­(EGMA-<i>co</i>-MaMA-SP) brushes, upon exposure to alternating visible and UV-light irradiation

Data_Sheet_1_A Compartmentalized Neuronal Cell-Culture Platform Compatible With Cryo-Fixation by High-Pressure Freezing for Ultrastructural Imaging.PDF

The human brain contains a wide array of billions of neurons and interconnections, which are often simplified for analysis in vitro using compartmentalized microfluidic devices for neuronal cell culturing, to better understand neuronal development and disease. However, such devices are traditionally incompatible for high-pressure freezing and high-resolution nanoscale imaging and analysis of their sub-cellular processes by methods including electron microscopy. Here we develop a novel compartmentalized neuronal co-culture platform allowing reconstruction of neuronal networks with high variable spatial control, which is uniquely compatible for high-pressure freezing. This cryo-fixation method is well-established to enable high-fidelity preservation of the reconstructed neuronal networks and their sub-cellular processes in a near-native vitreous state without requiring chemical fixatives. To direct the outgrowth of neurites originating from two distinct groups of neurons growing in the two different compartments, polymer microstructures akin to microchannels are fabricated atop of sapphire disks. Two populations of neurons expressing either enhanced green fluorescent protein (EGFP) or mCherry were grown in either compartment, facilitating the analysis of the specific interactions between the two separate groups of cells. Neuronally differentiated PC12 cells, murine hippocampal and striatal neurons were successfully used in this context. The design of this device permits direct observation of entire neuritic processes within microchannels by optical microscopy with high spatial and temporal resolution, prior to processing for high-pressure freezing and electron microscopy. Following freeze substitution, we demonstrate that it is possible to process the neuronal networks for ultrastructural imaging by electron microscopy. Several key features of the embedded neuronal networks, including mitochondria, synaptic vesicles, axonal terminals, microtubules, with well-preserved ultrastructures were observed at high resolution using focused ion beam – scanning electron microscopy (FIB-SEM) and serial sectioning – transmission electron microscopy (TEM). These results demonstrate the compatibility of the platform with optical microscopy, high-pressure freezing and electron microscopy. The platform can be extended to neuronal models of brain disease or development in future studies, enabling the investigation of subcellular processes at the nanoscale within two distinct groups of neurons in a functional neuronal pathway, as well as pharmacological testing and drug screening.</p

Poly(methyl methacrylate)-Based Nanofluidic Device for Rapid and Multiplexed Serological Antibody Detection of SARS-CoV‑2

The outbreak of SARS-CoV-2 has emphasized the value of point-of-care diagnostics, as well as reliable and cost-effective serological antibody tests, to monitor the viral spread and contain pandemics and endemics. Here, we present a three-dimensional (3D) nanofluidic device for rapid and multiplexed detection of viral antibodies. The device is made from poly­(methyl methacrylate) and contains 3D fluidic channels with nanoscale topography variations on the millimeter length scale, enabled by combining gray-scale e-beam lithography and nanoimprint lithography. It works with capillary pumps only and does not require a complex microfluidic setup and pumps, which hinder the widespread usage of micro- and nanofluidic devices. The device is designed to size dependently immobilize particles from a multiparticle mixture at predefined positions in nanochannels, resulting in distinct trapping lines. We show that it can be used as an on-chip fluorescence-linked immunosorbent assay for highly specific and sensitive multiplexed detection of serological antibodies against different viral proteins. Further test flexibility is demonstrated by on-bead color multiplexing for simultaneous detection of IgG and IgM antibodies in convalescent human serum. The particle sorting is further leveraged to enable concurrent detection of anti-spike (SARS-CoV-2) and anti-hemagglutinin (influenza A) antibodies. The device’s applications can be further extended to detect a large variety of diseases simultaneously in a reliable and straightforward manner