microcontact printing

The development and application of micropatterning technology play a promising role in the manipulation of biological substances and the exploration of life sciences at the microscale. However, the universally adaptable micropatterning method with user-friendly properties for acceptance in routine laboratories remains scarce. Herein, a green, facile, and rapid microcontact printing method is reported for upgrading popularization and diversification of biological patterning. The three-step printing can achieve high simplicity and fidelity of additive-free polydimethylsiloxane (PDMS) micropatterning and chip fabrication within 8 min as well as keep their high stability and diversity...

With both foodborne illness and food spoilage detrimentally impacting human health and the economy, there is growing interest in the development of in situ sensors that offer real-time monitoring of food quality within enclosed food packages. While oligonucleotide-based fluorescent sensors have illustrated significant promise, the development of such on-food sensors requires consideration towards sensing-relevant fluorescence properties of target food products-information that has not yet been reported. To address this need, comprehensive fluorescence profiles for various contamination-prone food products are established in this study across several wavelengths and timepoints...

Microcontact printing (MCP) is used to pattern a surface with a specific compound, allowing the spatially restricted response of cells to be assayed as they encounter a molecule of interest. MCP is a relatively low-cost and accessible technique that uses commercially available reagents and common cell culture equipment. However, it can be technically challenging, slow, and incompatible with microwell cell culture plates that are widely used for screening and other applications. Here, we describe a novel protocol using medical biopsy punches to transfer patterns into standard 96-well plates via polydimethylsiloxane (PDMS) cutouts...

Cells sense mechanical signals from the surrounding environment and transmit them to the nucleus through mechanotransduction to regulate cellular behavior. Microcontact printing (μCP), which utilizes elastomer stamps, is an effective method for simulating the cellular microenvironment and manipulating cell morphology. However, the conventional fabrication process of silicon masters and elastomer stamps requires complex procedures and specialized equipment, which restricts the widespread application of micropatterning in cell biology and hinders the investigation of the role of cell geometry in regulating cell behavior...

In the last decade human iPSC-derived cardiomyocytes (hiPSC-CMs) proved to be valuable for cardiac disease modeling and cardiac regeneration, yet challenges with scale, quality, inter-batch consistency, and cryopreservation remain, reducing experimental reproducibility and limiting clinical translation. Here, we report a robust cardiac differentiation protocol that uses Wnt modulation and a stirred suspension bioreactor to produce on average 124 million hiPSC-CMs with >90% purity using a variety of hiPSC lines (19 differentiations; 10 iPSC lines)...

INTRODUCTION: In native heart tissue, functions of cardiac fibroblasts (CFs) include synthesis, remodeling, and degradation of the extracellular matrix (ECM) as well as secreting factors that regulate cardiomyocyte (CM) function. The influence of direct co-culture and CF-derived ECM on CM mechanical function are not fully understood. METHODS: Here we use an engineered culture platform that provides control over ECM geometry and substrate stiffness to evaluate the influence of iPSC-CFs, and the ECM they produce, on the mechanical function of iPSC-CMs...

In Alzheimer's disease, the blood-brain barrier breakdown, blood vessel damage and re-organization are early events. Deposits of the small toxic peptide beta-amyloid (Aβ) cause the formation of extracellular plaques and accumulate in vessels disrupting the blood flow but may also play a role in blood clotting. In the present study, we aim to explore the impact of Aβ on the migration of endothelial cells and subsequent vessel formation. We use organotypic brain slices of postnatal day 10 wildtype mice (C57BL/6) and connect them to small microcontact prints (µCPs) of collagen...

Parallel lithographic deposition of polymers onto counterpart substrates is a widely applied surface manufacturing operation. However, polymers may only be soluble in organic solvents or are insoluble at all. Solvent evaporation during stamping may trigger hardly controllable capillarity-driven flow processes or phase separation, and polymer solutions may spread on the counterpart substrates. Solvent-free stamping of melts prevents these drawbacks. Here, a stamp design for the deposition of melts is devised, which intrinsically circumvents ink depletion...

Multicellular three-dimensional (3D) in vitro models, such as cell spheroids and organoids, can significantly improve the viability, histomorphology, genotype stability, function and drug metabolism of cells [1], [2], [3]. In general, several culture methods of 3D models, including the hanging drop, microwell-mesh and hydrogel encapsulating methods, have difficulty building a standard mode and controlling the size and arrangement of cell spheroids or organoids, which could severely affect the authenticity and repeatability of experimental results [4]...

The present study investigates silicone transfer occurring during microcontact printing (μCP) of lectins with polydimethylsiloxane (PDMS) stamps and its impact on the adhesion of cells. Static adhesion assays and single-cell force spectroscopy (SCFS) are used to compare adhesion of nonmalignant (HCV29) and cancer (HT1376) bladder cells, respectively, to high-affinity lectin layers (PHA-L and WGA, respectively) prepared by physical adsorption and μCP. The chemical composition of the μCP lectin patterns was monitored by time-of-flight secondary ion mass spectrometry (ToF-SIMS)...

In recent years, innovative cell-based biosensing systems have been developed, showing impact in healthcare and life science research. Now, there is a need to design mass-production processes to enable their commercialization and reach society. However, current protocols for their fabrication employ materials that are not optimal for industrial production, and their preparation requires several chemical coating steps, resulting in cumbersome protocols. We have developed a simplified two-step method for generating controlled cell patterns on PMMA, a durable and transparent material frequently employed in the mass manufacturing of microfluidic devices...

Surfactants and other amphiphilic molecules are used extensively in household products, industrial processes, and biological applications and are also common environmental contaminants; as such, methods that can detect, sense, or quantify them are of great practical relevance. Aqueous emulsions of thermotropic liquid crystals (LCs) can exhibit distinctive optical responses in the presence of surfactants and have thus emerged as sensitive, rapid, and inexpensive sensors or reporters of environmental amphiphiles...

Highly sensitive and selective choline microbiosensors were constructed by microcontact printing (μCP) of choline oxidase (ChOx) in a crosslinked, polyamine-functionalized zwitterionic polymer matrix on microelectrode arrays (MEAs). μCP has emerged as a potential means to create implantable, multiplexed sensor microprobes, which requires the targeted deposition of different sensor materials to specific microelectrode sites on a MEA. However, the less than sufficient enzyme loading and inadequate spatial resolution achieved with current μCP approaches has limited adoption of the method for electroenzymatic microsensors...

Integrin-dependent adhesion to the extracellular matrix (ECM) mediates mechanosensing and signaling in response to altered microenvironmental conditions. In order to provide tissue- and organ-specific cues, the ECM is composed of many different proteins that temper the mechanical properties and provide the necessary structural diversity. Despite most human tissues being soft, the prevailing view from predominantly in vitro studies is that increased stiffness triggers effective cell spreading and activation of mechanosensitive signaling pathways...

Organotypic brain slices are three-dimensional 150 µm-thick sections of a postnatal day 10 mouse and can be cultured for several weeks in vitro. In such brain slices the complex cellular connections are preserved with a high viability. These brain slices can be connected to collagen-loaded microcontact prints to develop a simple brain-on-a-chip model. Using the microcontact printing technique, many peptides or proteins can be printed onto a semipermeable membrane and linked to brain slices. On these microcontact prints, brain-derived nerve fibers grow out, or microglia can get activated and migrate out, or also new brain vessels can be formed...

Microcontact printing (CP) is widely used to guide neurons to form 2D networks for neuroscience research. However, it is still difficult to establish 3D neuronal cultures on the CP substrate even though 3D neuronal structures are able to recapitulate critical aspects of native tissue. Here, we demonstrate that the reduced cell-substrate adhesion caused by the CP substrate could conveniently facilitate the aggregate formation of large-scale 3D neuron cluster networks. Furthermore, based on the quantitative analysis of the calcium activity of the resulting cluster networks, the effect of cell seeding density and local restriction of the CP substrate on network dynamics was investigated in detail...

The recent advancements of single-cell analysis have significantly enhanced the ability to understand cellular physiology when compared to bulk cellular analysis. Here a massively parallel single-cell patterning and very large biomolecular delivery is reported. Micro-pillar polydimethyl siloxane stamp with different diameters (40-100 µm with 1 cm × 1 cm patterning area) is fabricated and then imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters...

N-Heterocyclic carbene (NHC)-modified planar gold surfaces (NHC@Au) were found to be more susceptible toward wet chemical etching than undecorated surface areas. Site-selective decoration of NHCs on Au was achieved by microcontact printing (μCP) of the NHC precursors 1,3-bis(diisopropylphenyl)imidazol-3-ium hydrogen carbonate (IPr(H)[HCO3 ]) or 1,3-dimethylbenzimidazol-3-ium hydrogen carbonate (BIMe(H)[HCO3 ]). Strikingly, BIMe@Au showed concentration-dependent etching behavior, tunable from a positive resist to a negative resist...

Muscle tissue engineering has been the focus of extensive research due to its potential for numerous medical applications including ex vivo actuator development and clinical treatments. In this study, we developed a method for harvesting muscle fiber in a floatable and translocatable manner utilizing thermally expandable hydrogels with a chemically-patterned polydopamine (PD) layer generated by microcontact printing (μCP). The μCP of PD on the hydrogel facilitated the formation of stripe patterns with varying widths of printed/non-printed area (50/50, 100/100, and 200/200 μm)...

Polymeric surface coatings capable of effectively integrating desired functional molecules and ligands are attractive for fabricating bio-interfaces necessary for various applications. Herein, we report the design of a polymeric platform amenable to such modifications in a modular fashion through host-guest chemistry. Copolymers containing adamantane (Ada) moieties, diethylene glycol (DEG) units, and silyloxy groups to provide functionalization handles, anti-biofouling character, and surface attachment, respectively, were synthesized...