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Nuclear magnetic resonance (NMR) spectroscopy is commonly employed in a wide range of metabolomic research. Unfortunately, due to its relatively low sensitivity, smaller samples become challenging to study by NMR. Cryoprobes can be used to increase sensitivity by cooling the coil and preamplifier, offering sensitivity improvements of â¼3 to 4x. Alternatively, microcoils can be used to increase mass sensitivity by improving sample filling and proximity, along with decreased electrical resistance. Unfortunately, combining the two approaches is not just technically challenging, but as the coil decreases, so does its thermal fingerprint, reducing the advantage of cryogenic cooling. Here, an alternative solution is proposed in the form of a Lenz lens inside a cryoprobe. Rather than replacing the detection coil, Lenz lenses allow the B1 field from a larger coil to be refocused onto a much smaller sample area. In turn, the stronger B1 field at the sample provides strong coupling to the cryocoil, improving the signal. By combining a 530 I.D. Lenz lens with a cryoprobe, sensitivity was further improved by 2.8x and 3.5x for 1H and 13C, respectively, over the cryoprobe alone for small samples. Additionally, the broadband nature of the Lenz lenses allowed multiple nuclei to be studied and heteronuclear two-dimensional (2D) NMR approaches to be employed. The sensitivity improvements and 2D capabilities are demonstrated on 430 nL of hemolymph and eight eggs (â¼350 µm O.D.) from the model organismDaphnia magna. In summary, combining Lenz lenses with cryoprobes offers a relatively simple approach to boost sensitivity for tiny samples while retaining cryoprobe advantages.
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Lentes , Imageamento por Ressonância Magnética , Animais , Espectroscopia de Ressonância Magnética/métodos , Imageamento por Ressonância Magnética/métodos , Temperatura Baixa , Monitoramento AmbientalRESUMO
Microfluidic water-in-oil droplets are a versatile tool for biological and biochemical applications due to the advantages of extremely small monodisperse reaction vessels in the pL-nL range. A key factor for the successful dissemination of this technology to life science laboratory users is the ability to produce microfluidic droplet generators and related accessories by low-entry barrier methods, which enable rapid prototyping and manufacturing of devices with low instrument and material costs. The direct, experimental side-by-side comparison of three commonly used additive manufacturing (AM) methods, namely fused deposition modeling (FDM), inkjet printing (InkJ), and stereolithography (SLA), is reported. As a benchmark, micromilling (MM) is used as an established method. To demonstrate which of these methods can be easily applied by the non-expert to realize applications in topical fields of biochemistry and microbiology, the methods are evaluated with regard to their limits for the minimum structure resolution in all three spatial directions. The suitability of functional SLA and MM chips to replace classic SU-8 prototypes is demonstrated on the basis of representative application cases.
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Disciplinas das Ciências Biológicas , Dispositivos Lab-On-A-Chip , Enzimas/metabolismo , Desenho de Equipamento , Cinética , Impressão Tridimensional , EstereolitografiaRESUMO
BACKGROUND: Biomedical diagnostic and lab automation solutions built on the Lab-on-a-Disc (LoaD) platform has great potential due to their independence from specialised micro-pumps and their ease of integration, through direct pipetting, with manual or automated workflows. However, a challenge for all microfluidic chips is their cost of manufacture when each microfluidic disc must be customized for a specific application. In this paper, we present centrifugal discs with programmable fluidic networks. RESULTS: Based on dissolvable film valves, we present two technologies. The first, based on recently introduced pulse-actuated dissolvable film valves, is a centrifugal disc which, depending on how it is loaded, is configured to perform either six sequential reagent releases through one reaction chamber or three sequential reagent releases through two reaction chambers. In the second approach, we use the previously introduced electronic Lab-on-a-Disc (eLoaD) wireless valve array, which can actuate up to 128 centrifugo-pneumatic dissolvable film valves in a pre-defined sequence. In this approach we present a disc which can deliver any one of 8 reagent washes to any one of four reaction chambers. We use identical discs to demonstrate the first four sequential washes through two reaction chambers and then two sequential washes through four reaction chambers. SIGNIFICANCE: These programmable fluidic networks have the potential to allow a single disc architecture to be applied to multiple different assay types and so can offer a lower-cost and more integrated alternative to the standard combination of micro-titre plate and liquid handling robot. Indeed, it may even be possible to conduct multiple different assays concurrently. This can have the effect of reducing manufacturing costs and streamlining supply-chains and so results in a more accessible diagnostic platform.
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Laser-induced graphene (LIG) has been emerging as a promising electrode material for supercapacitors due to its cost-effective and straightforward fabrication approach. However, LIG-based supercapacitors still face challenges with limited capacitance and stability. To overcome these limitations, in this work, we present a novel, cost-effective, and facile fabrication approach by integrating LIG materials with candle-soot nanoparticles. The composite electrode is fabricated by laser irradiation on a Kapton sheet to generate LIG material, followed by spray-coating with candle-soot nanoparticles and annealing. Materials characterization reveals that the annealing process enables a robust connection between the nanoparticles and the LIG materials and enhances nanoparticle graphitization. The prepared supercapacitor yields a maximum specific capacitance of 15.1 mF/cm2 at 0.1 mA/cm2, with a maximum energy density of 2.1 µWh/cm2 and a power density of 50 µW/cm2. Notably, the synergistic activity of candle soot and LIG surpasses the performances of previously reported LIG-based supercapacitors. Furthermore, the cyclic stability of the device demonstrates excellent capacitance retention of 80% and Coulombic efficiency of 100% over 10000 cycles.
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The integration of additive manufacturing technologies with the pyrolysis of polymeric precursors enables the design-controlled fabrication of architected 3D pyrolytic carbon (PyC) structures with complex architectural details. Despite great promise, their use in cellular interaction remains unexplored. This study pioneers the utilization of microarchitected 3D PyC structures as biocompatible scaffolds for the colonization of muscle cells in a 3D environment. PyC scaffolds are fabricated using micro-stereolithography, followed by pyrolysis. Furthermore, an innovative design strategy using revolute joints is employed to obtain novel, compliant structures of architected PyC. The pyrolysis process results in a pyrolysis temperature- and design-geometry-dependent shrinkage of up to 73%, enabling the geometrical features of microarchitected compatible with skeletal muscle cells. The stiffness of architected PyC varies with the pyrolysis temperature, with the highest value of 29.57 ± 0.78 GPa for 900 °C. The PyC scaffolds exhibit excellent biocompatibility and yield 3D cell colonization while culturing skeletal muscle C2C12 cells. They further induce good actin fiber alignment along the compliant PyC construction. However, no conclusive myogenic differentiation is observed here. Nevertheless, these results are highly promising for architected PyC scaffolds as multifunctional tissue implants and encourage more investigations in employing compliant architected PyC structures for high-performance tissue engineering applications.
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Engenharia Tecidual , Alicerces Teciduais , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Carbono , Células Musculares , Impressão TridimensionalRESUMO
Extreme point-of-care refers to medical testing in unfavorable conditions characterized by a lack of primary resources or infrastructure. As witnessed in the recent past, considerable interest in developing devices and technologies exists for extreme point-of-care applications, for which the World Health Organization has introduced a set of encouraging and regulating guidelines. These are referred to as the ASSURED criteria, an acronym for Affordable (A), Sensitive (S), Specific (S), User friendly (U), Rapid and Robust (R), Equipment-free (E), and Delivered (D). However, the current extreme point of care devices may require an intermediate sample preparation step for performing complex biomedical analysis, including the diagnosis of rare-cell diseases and early-stage detection of sepsis. This article assesses the potential of carbon-electrode dielectrophoresis (CarbonDEP) for sample preparation competent in extreme point-of-care, following the ASSURED criteria. We first discuss the theory and utility of dielectrophoresis (DEP) and the advantages of using carbon microelectrodes for this purpose. We then critically review the literature relevant to the use of CarbonDEP for bioparticle manipulation under the scope of the ASSURED criteria. Lastly, we offer a perspective on the roadmap needed to strengthen the use of CarbonDEP in extreme point-of-care applications.
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Carbon materials constitute a growing family of high-performance materials immersed in ongoing scientific technological revolutions. Their biochemical properties are interesting for a wide set of healthcare applications and their biomechanical performance, which can be modulated to mimic most human tissues, make them remarkable candidates for tissue repair and regeneration, especially for articular problems and osteochondral defects involving diverse tissues with very different morphologies and properties. However, more systematic approaches to the engineering design of carbon-based cell niches and scaffolds are needed and relevant challenges should still be overcome through extensive and collaborative research. In consequence, this study presents a comprehensive description of carbon materials and an explanation of their benefits for regenerative medicine, focusing on their rising impact in the area of osteochondral and articular repair and regeneration. Once the state-of-the-art is illustrated, innovative design and fabrication strategies for artificially recreating the cellular microenvironment within complex articular structures are discussed. Together with these modern design and fabrication approaches, current challenges, and research trends for reaching patients and creating social and economic impacts are examined. In a closing perspective, the engineering of living carbon materials is also presented for the first time and the related fundamental breakthroughs ahead are clarified.
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Cartilagem Articular , Engenharia Tecidual , Carbono , Humanos , Alicerces TeciduaisRESUMO
Carbon origami enables the fabrication of lightweight and mechanically stiff 3D complex architectures of carbonaceous materials, which have a high potential to impact a wide range of applications positively. The precursor materials and their inherent microstructure play a crucial role in determining the properties of carbon origami structures. Here, non-porous polyaramid Nomex sheets and macroporous fibril cellulose sheets are explored as the precursor sheets for studying the effect of precursor nature and microstructure on the material and structural properties of the carbon origami structures. The fabrication process involves pre-creasing precursor sheets using a laser engraving process, followed by manual-folding and carbonization. The cellulose precursor experiences a severe structural shrinkage due to its macroporous fibril morphology, compared to the mostly non-porous morphology of Nomex-derived carbon. The morphological differences further yield a higher specific surface area for cellulose-derived carbon. However, Nomex results in more crystalline carbon than cellulose, featuring a turbostratic microstructure like glassy carbon. The combined effect of morphology and glass-like features leads to a high mechanical stiffness of 1.9 ± 0.2 MPa and specific modulus of 2.4 × 104 m2·s-2 for the Nomex-derived carbon Miura-ori structure, which are significantly higher than cellulose-derived carbon Miura-ori (elastic modulus = 504.7 ± 88.2 kPa; specific modulus = 1.2 × 104 m2·s-2) and other carbonaceous origami structures reported in the literature. The results presented here are promising to expand the material library for carbon origami, which will help in the choice of suitable precursor and carbon materials for specific applications.
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This work establishes carbon nanofibre-mediated patterning of metal oxide nanostructures, through the combination of electrospinning and vapor-phase transport growth. Electrospinning of a suitable precursor with subsequent carbonization results in the patterning of catalyst gold nanoparticles embedded within carbon nanofibres. During vapor-phase transport growth, these nanofibres allow preferential growth of one-dimensional metal oxide nanostructures, which grow radially outward from the nanofibril axis, yielding a hairy caterpillar-like morphology. The synthesis of metal oxide caterpillars is demonstrated using zinc oxide, indium oxide, and tin oxide. Source and substrate temperatures play the most crucial role in determining the morphology of the metal oxide caterpillars, whereas the distribution of the nanofibres also has a significant impact on the overall morphology. Introducing the current methodology with near-field electrospinning further facilitates user-defined custom patterning of metal oxide caterpillar-like structures.
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It is generally accepted that inducing molecular alignment in a polymer precursor via mechanical stresses influences its graphitization during pyrolysis. However, our understanding of how variations of the imposed mechanics can influence pyrolytic carbon microstructure and functionality is inadequate. Developing such insight is consequential for different aspects of carbon MEMS manufacturing and applicability, as pyrolytic carbons are the main building blocks of MEMS devices. Herein, we study the outcomes of contrasting routes of stress-induced graphitization by providing a comparative analysis of the effects of compressive stress versus standard tensile treatment of PAN-based carbon precursors. The results of different materials characterizations (including scanning electron microscopy, Raman and X-ray photoelectron spectroscopies, as well as high-resolution transmission electron microscopy) reveal that while subjecting precursor molecules to both types of mechanical stresses will induce graphitization in the resulting pyrolytic carbon, this effect is more pronounced in the case of compressive stress. We also evaluated the mechanical behavior of three carbon types, namely compression-induced (CIPC), tension-induced (TIPC), and untreated pyrolytic carbon (PC) by Dynamic Mechanical Analysis (DMA) of carbon samples in their as-synthesized mat format. Using DMA, the elastic modulus, ultimate tensile strength, and ductility of CIPC and TIPC films are determined and compared with untreated pyrolytic carbon. Both stress-induced carbons exhibit enhanced stiffness and strength properties over untreated carbons. The compression-induced films reveal remarkably larger mechanical enhancement with the elastic modulus 26 times higher and tensile strength 2.85 times higher for CIPC compared to untreated pyrolytic carbon. However, these improvements come at the expense of lowered ductility for compression-treated carbon, while tension-treated carbon does not show any loss of ductility. The results provided by this report point to the ways that the carbon MEMS industry can improve and revise the current standard strategies for manufacturing and implementing carbon-based micro-devices.
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Multiscale 3D carbon architectures are of particular interest in tissue engineering applications, as these structures may allow for three-dimensional cell colonization essential for tissue growth. In this work, carbon fiber/microlattice hybrid architectures are introduced as innovative multi-scale scaffolds for tissue engineering. The microlattice provides the design freedom and structural integrity, whereas the fibrous component creates a cellular microenvironment for cell colonization. The hybrid structures are fabricated by carbonization of stereolithographically 3D printed epoxy microlattice architectures which are pre-filled with cotton fibers within the empty space of the architectures. The cotton filling result in less shrinkage of the architecture during carbonization, as the tight confinement of the fibrous material prevents the free-shrinkage of the microlattices. The hybrid architecture exhibits a compressive strength of 156.9±25.6 kPa, which is significantly higher than an empty carbon microlattice architecture. Furthermore, the hybrid architecture exhibits a flexible behavior up to 30% compressive strain, which is also promising towards soft-tissue regeneration. Osteoblast-like murine MC3T3-E1 cells are cultured within the 3D hybrid structures. Results show that the cells are able to not only proliferate on the carbon microlattice elements as well as along the carbon fibers, but also make connections with each other across the inner pores created by the fibers, leading to a three-dimensional cell colonization. These carbon fiber/microlattice hybrid structures are promising for future fabrication of functionally graded scaffolds for tissue repair applications.
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Engenharia Tecidual , Alicerces Teciduais , Animais , Fibra de Carbono , Força Compressiva , Camundongos , Osteoblastos , Impressão TridimensionalRESUMO
A novel approach for automated high throughput NMR spectroscopy with improved mass-sensitivity is accomplished by integrating microfluidic technologies and micro-NMR resonators. A flow system is utilized to transport a sample of interest from outside the NMR magnet through the NMR detector, circumventing the relatively vast dead volume in the supplying tube by loading a series of individual sample plugs separated by an immiscible fluid. This dual-phase flow demands a real-time robust sensing system to track the sample position and velocities and synchronize the NMR acquisition. In this contribution, we describe an NMR probe head that possesses a microfluidic system featuring: (i) a micro saddle coil for NMR spectroscopy and (ii) a pair of interdigitated capacitive sensors flanking the NMR detector for continuous position and velocity monitoring of the plugs with respect to the NMR detector. The system was successfully tested for automating flow-based measurement in a 500 MHz NMR system, enabling high resolution spectroscopy and NMR sensitivity of 2.18 nmol s1/2 with the flow sensors in operation. The flow sensors featured sensitivity to an absolute difference of 0.2 in relative permittivity, enabling distinction between most common solvents. It was demonstrated that a fully automated NMR measurement of nine individual 120 µL samples could be done within 3.6 min or effectively 15.3 s per sample.
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Within microfluidic technologies, the centrifugal microfluidic "Lab-on-a-Disc" (LoaD) platform offers great potential for use at the PoC and in low-resource settings due to its robustness and the ability to port and miniaturize 'wet bench' laboratory protocols. We present the combination of 'event-triggered dissolvable film valves' with a centrifugo-pneumatic siphon structure to enable control and timing, through changes in disc spin-speed, of the release and incubations of eight samples/reagents/wash buffers. Based on these microfluidic techniques, we integrated and automated a chemiluminescent immunoassay for detection of the CVD risk factor marker C-reactive protein displaying a limit of detection (LOD) of 44.87 ng mL-1 and limit of quantitation (LoQ) of 135.87 ng mL-1.
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Proteína C-Reativa/análise , Dispositivos Lab-On-A-Chip , Automação , Biomarcadores/análise , Doenças Cardiovasculares/metabolismo , Desenho de Equipamento , Humanos , Laboratórios , Limite de Detecção , Técnicas Analíticas Microfluídicas , MicrofluídicaRESUMO
Improvements to the signal-to-noise ratio of magnetic resonance detection lead to a strong reduction in measurement time, yet as a sole optimization goal for resonator design, it would be an oversimplification of the problem at hand. Multiple constraints, for example for field homogeneity and sample shape, suggest the use of numerical optimization to obtain resonator designs that deliver the intended improvement. Here we consider the 2D Lenz lens to be a sufficiently broadband flux transforming interposer between the sample and a radiofrequency (RF) circuit and to be a flexible and easily manufacturable device family with which to mediate different design requirements. We report on a method to apply topology optimization to determine the optimal layout of a Lenz lens and demonstrate realizations for both low- (45â¯MHz) and high-frequency (500â¯MHz) nuclear magnetic resonance.
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We introduce a low-complexity, low-cost, yet sufficiently accurate automatic tune and match system for NMR and MRI applications. The ArduiTaM builds upon an Arduino Uno embedded system that drives a commercial frequency synthesiser chip to perform a frequency sweep around the Larmor frequency. The generated low-power signal is fed to the NMR coil, after which the reflected waves are detected using a directional coupler and amplified. The signal shape is then extracted by means of an envelope detector and passed on to the Arduino, which performs a dip search while continuously generating actuator control patterns to adjust the tune and match capacitors. The process stops once the signal dip reaches the Larmor frequency. The ArduiTaM works readily with any spectrometer frequency in the range from 1 to 23â¯T. The speed of the ArduiTaM is mainly limited by the clock of the Arduino and the capacitor actuation mechanism. The Arduino can easily be replaced by a higher-speed microcontroller, and varactors can replace stepper-motor controlled variable capacitors. The ArduiTaM is made available in open source, and so is easily duplicated.
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Edible rice paper wrapper is found to be an interesting precursor of a porous and light-weight carbon material. During pyrolysis, material samples show significant differences in length change, displaying typical 20-25% shrinking in the in-plane directions, and strongly expanding (up to 500%) across their out-of-plane direction. This results in a template-free synthesis of a 3D network of cellular carbon material. The out-of-plane expansion also allows for fabrication of 3D shapes of cellular carbon material from the 2D precursor. The rice paper derived carbon material features a hierarchical porosity, resulting in a specific surface area ranging from 6 m2 g-1 to 239 m2 g-1 depending on the synthesis temperature. The carbon material has a density of 0.02-0.03 g cm-3, and a higher modulus-density ratio than reported for other cellular carbon materials. It is mechanically stiff and exhibits excellent fire-resistant properties.
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A method for the fabrication of flexible electrical circuits on polyaramid substrates is presented based on laser-induced carbonization followed by copper electroplating. Locally carbonized flexible sheets of polyaramid (Nomex), by laser radiation, create rough and highly porous microstructures that show a higher degree of graphitization than thermally carbonized Nomex sheets. The found recipe for laser-induced carbonization creates conductivities of up to â¼45 S cm-1, thereby exceeding that observed for thermally pyrolyzed materials (â¼38 S cm-1) and laser carbon derived from Kapton using the same laser wavelength (â¼35 S cm-1). The electrical conductivity of the carbonized tracks was further improved by electroplating with copper. To demonstrate the electrical performance, fabricated circuits were tested and improvement of the sheet resistance was determined. Copper films exhibit antimicrobial activity and were used to fabricate customized flexible antibacterial coatings. The integration of laser carbonization and electroplating technologies in a polyaramid substrate points to the development of customized circuit designs for smart textiles operating in high-temperature environments.
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Antibacterianos/química , Cobre/química , Lasers , Nylons/química , Antibacterianos/farmacologia , Bacillus subtilis/efeitos dos fármacos , Carbono/química , Cobre/farmacologia , Galvanoplastia , Escherichia coli/efeitos dos fármacosRESUMO
A scalable, generic wireless colour detector for point-of-care diagnostics in resource-limited settings is presented. The challenges faced in these settings have limited the effectiveness of point-of-care diagnostics. By combining the growing fields of paper-based diagnostics and printed electronics with Southern African clinic perspectives, a mass-producible, low-cost, paper-based solution for result readout and communication was developed. Printed radio frequency identification devices with sensing capabilities were manufactured, targeting colour detection from lateral flow test strip devices and other typical paper-based rapid test formats. The results were compared to those obtained from a commercial lateral flow test strip reader and image analysis using ImageJ, and demonstrate suitability for delivering automated readout and communication of results. The wireless colour detector is compatible with different test strip form factors, providing a modular solution and reducing the need for training. The solution is low cost and maintenance free, and thus fitting for resource-limited settings. A scalable version of the solution has been developed, making use of standard manufacturing processes for printing and packaging industries, initially using sheet-to-sheet formats, but with the goal of being scalable to roll-to-roll processes. This would enable the possibility of local manufacture, and mass distribution of the devices to those resource-limited areas where they are most needed, and where they will have the greatest impact on point-of-care testing.
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Técnicas Biossensoriais , Colorimetria , Técnicas Eletroquímicas , Testes Imediatos , Técnicas Biossensoriais/instrumentação , Colorimetria/instrumentação , Técnicas Eletroquímicas/instrumentação , HumanosRESUMO
Multilayer flexible substrates offer a means to combine high lateral precision and resolution with roll-up processes, allowing layer-based manufacturing to reach into the third dimension. Here we explore this combination to achieve an otherwise hard-to-manufacture resonator geometry: the double-helix. The use of commercial flexPCB technology enabled optimal winding connections and a versatile adjustment to various operation fields, sample volumes and resonance numbers. The sensitivity of the design is shown to greatly benefit from the fabrication method, though optimal electrical connections and several radially-wound windings, and was measured to outperform an equivalent solenoid despite the known geometrical disadvantage.
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A simple, efficient, and repeatable combination of wax printing and hot embossing is reported. This combination yields microfluidic channels in paper, where fluid transport driven by paper-intrinsic capillary forces takes place inside the noncompressed areas, whereas embossed and wax-bonded areas serve as hydrophobic barriers laterally confining the fluid flow. Lab-made paper sheets first coated with a hydrophobic wax were hot-embossed with a tailor-made metal stamp. Both paper-intrinsic (e.g., grammage, fiber type) and paper-extrinsic parameters (e.g., embossing force) were studied for their influence on the geometry of the embossed structures and the resulting redistribution of the wax within the paper matrix. Embossing of wax-printed paper at temperatures above the wax melting point was completed within 15 s. Cotton linters papers required higher embossing forces than eucalyptus papers, which can be explained by their different intrinsic mechanical properties. In summary, both paper-intrinsic and paper-extrinsic parameters were found to have strong impact on resolution and reproducibility of the channels. All in all, the approach yields microfluidic channels in a fast and robust and reproducible manner with comparably low constrains on the precision of manufacturing parameters, such as embossing time, force, or temperature. Most importantly, embossing greatly reduces the lateral spreading of the wax as seen with melting approaches and therefore allows for a much higher feature density than the latter.