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1.
The lack of technologies that combine automated manipulation, sorting, as well as immobilization of single metazoan embryos remains the key obstacle to high-throughput organism-based ecotoxicological analysis and drug screening routines. Noticeably, the major obstacle hampering the automated trapping and arraying of millimetre-sized embryos on chip-based devices is their substantial size and mass, which lead to rapid gravitational-induced sedimentation and strong inertial forces. In this work, we present a comprehensive mechanistic and design rationale for manipulation and passive trapping of individual zebrafish embryos using only hydrodynamic forces. We provide evidence that by employing innovative design features, highly efficient hydrodynamic positioning of large embryos on a chip can be achieved. We also show how computational fluid dynamics-guided design and the Lagrangian particle tracking modeling can be used to optimize the chip performance. Importantly, we show that rapid prototyping and medium scale fabrication of miniaturized devices can be greatly accelerated by combining high-speed laser prototyping with replica moulding in poly(dimethylsiloxane) instead of conventional photolithography techniques. Our work establishes a new paradigm for chip-based manipulation of large multicellular organisms with diameters well above 1 mm and masses often exceeding 1 mg. Passive docking of large embryos is an attractive alternative to provide high level of automation while alleviating potentially deleterious effects associated with the use of active chip actuation. This greatly expands the capabilities of bioanalyses performed on small model organisms and offers numerous and currently inaccessible laboratory automation advantages.  相似文献   

2.
Bio-electrosprays (BESs) provide a means of precisely manipulating cells and thus have the potential for many clinical uses such as the generation of artificial tissues∕organs. Previously we tested the biological safety of this technology with a variety of living cells and also embryos from the vertebrate model organisms Danio rerio (zebrafish) and Xenopus tropicalis (frog). However, the viability and fertility of the treated embryos could not be fully assessed due to animal licensing laws. Here we assay the viability and fertility of Drosophila melanogaster (fruit fly) embryos in conjunction with the bio-electrospray procedure. Bio-electrosprayed Drosophila embryos developed into fully fertile adult flies that were indistinguishable from wild-type. Thus, we demonstrate that the bio-electrospray procedure does not induce genetic or physical damage that significantly affects the development or fertility of a multicellular organism. This study along with our previous investigations demonstrates the potential of this approach to be developed for the precise manipulation of sensitive biological materials.  相似文献   

3.
This work presents an array of microfluidic chambers for on-chip culturing of microorganisms in static and continuous shear-free operation modes. The unique design comprises an in-situ polymerized hydrogel that forms gas and reagent permeable culture wells in a glass chip. Utilizing a hydrophilic substrate increases usability by autonomous capillary priming. The thin gel barrier enables efficient oxygen supply and facilitates on-chip analysis by chemical access through the gel without introducing a disturbing flow to the culture. Trapping the suspended microorganisms inside a gel well allows for a much simpler fabrication than in conventional trapping devices as the minimal feature size does not depend on cell size. Nutrients and drugs are provided on-chip in the gel for a self-contained and user-friendly handling. Rapid antibiotic testing in static cultures with strains of Enterococcus faecalis and Escherichia coli is presented. Cell seeding and diffusive medium supply is provided by phaseguide technology, enabling simple operation of continuous culturing with a great flexibility. Cells of Saccharomyces cerevisiae are utilized as a model to demonstrate continuous on-chip culturing.  相似文献   

4.
Investigations were conducted to determine acute effects of Pb2+ and Cu2+ presented individually and collectively on zebrafish embryos. Aquatic safety testing requires a cheap, fast, and highly efficient platform for real-time evaluation of single and mixture of metal toxicity. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic effects of Pb2+ and Cu2+ using zebrafish (Danio rerio) embryos. The microfluidic chip is composed of a disc-shaped concentration gradient generator and 24 culture chambers, which can generate one blank solution, seven mixture concentrations, and eight single concentrations for each metal solution, thus enabling the assessment of zebrafish embryos. To test the accuracy of this new chip platform, we have examined the toxicity and teratogenicity of Pb2+ and Cu2+ on embryos. The individual and combined impact of Pb2+ and Cu2+ on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators, such as spontaneous motion at 22 hours post fertilization (hpf), mortality at 24 hpf, heartbeat and body length at 96 hpf, etc. It was found that Pb2+ or Cu2+ could induce deformity and cardiovascular toxicity in zebrafish embryos and the mixture could induce more severe toxicity. This chip is a multiplexed testing apparatus that allows for the examination of toxicity and teratogenicity for substances and it also can be used as a potentially cost-effective and rapid aquatic safety assessment tool.  相似文献   

5.
In this work, we demonstrate the use of stereolithographic 3D printing to fabricate millifluidic devices, which are used to engineer particles with multiple compartments. As the 3D design is directly transferred to the actual prototype, this method accommodates 3D millimeter-scaled features that are difficult to achieve by either lithographic-based microfabrication or traditional macrofabrication techniques. We exploit this approach to produce millifluidic networks to deliver multiple fluidic components. By taking advantage of the laminar flow, the fluidic components can form liquid jets with distinct patterns, and each pattern has clear boundaries between the liquid phases. Afterwards, droplets with controlled size are fabricated by spraying the liquid jet in an electric field, and subsequently converted to particles after a solidification step. As a demonstration, we fabricate calcium alginate particles with structures of (1) slice-by-slice multiple lamellae, (2) concentric core-shells, and (3) petals surrounding the particle centers. Furthermore, distinct hybrid particles combining two or more of the above structures are also obtained. These compartmentalized particles impart spatially dependent functionalities and properties. To show their applicability, various ingredients, including fruit juices, drugs, and magnetic nanoparticles are encapsulated in the different compartments as proof-of-concepts for applications, including food, drug delivery, and bioassays. Our 3D printed electro-millifluidic approach represents a convenient and robust method to extend the range of structures of functional particles.  相似文献   

6.
Microfluidic technologies have several advantages in sample preparation for diagnostics but suffer from the need for an external operation system that hampers user-friendliness. To overcome this limitation in microfluidic technologies, a number of user-friendly methods utilizing capillary force, degassed poly(dimethylsiloxane), pushbutton-driven pressure, a syringe, or a pipette have been reported. Among these methods, the pushbutton-driven, pressure-based method has a great potential to be widely used as a user-friendly sample preparation tool for point-of-care testing or portable diagnostics. In this Perspective, we focus on the pushbutton-activated microfluidic technologies toward a user-friendly sample preparation tool. The working principle and recent advances in pushbutton-activated microfluidic technologies are briefly reviewed, and future perspectives for wide application are discussed in terms of integration with the signal analysis system, user-dependent variation, and universal and facile use.  相似文献   

7.
90 kDa heat shock protein (HSP90) is a ubiquitous molecular chaperone and is one of the abundant proteins present in a cell under normal and stressed conditions. The adenosine triphosphate (ATP) binding region of HSP90 is currently under a great degree of study because of the interest of its role in cancer and protein maintenance; the binding of ATP to HSP90 induces a large conformational change in the protein as a result of the activity of different types of stressors within the cells. In the present paper, a simple microfluidic biosensor is proposed for the characterization of ATP-HSP90 interactions through the principle of bioresistive variation. The experimental results prove that the present biosensor system is highly suitable for the detection of heat shock proteins present in a real-time biological sample, which is very useful for in-situ biomedical applications and rapid pathogenic detections.  相似文献   

8.
Additive manufacturing was adopted in multiple fields of life sciences. It is also becoming a popular tool for rapid prototyping of microfluidic and biomedical devices. Limited studies have been performed to investigate the biological implications of using 3D printed polymers. Here we assessed the biocompatibility of seven commercially available polymers, using a battery of standardized bioassays for chemical risk assessment. Our data show that leachates from photopolymers substrata appear to be very toxic to vertebrates and several invertebrate indicator organisms. These results demonstrate significant consequences for the use of selected photopolymers in the fabrication of bio-devices.  相似文献   

9.
The capability of 3D printing technologies for direct production of complex 3D structures in a single step has recently attracted an ever increasing interest within the field of microfluidics. Recently, ultrafast lasers have also allowed developing new methods for production of internal microfluidic channels within the bulk of glass and polymer materials by direct internal 3D laser writing. This review critically summarizes the latest advances in the production of microfluidic 3D structures by using 3D printing technologies and direct internal 3D laser writing fabrication methods. Current applications of these rapid prototyped microfluidic platforms in biology will be also discussed. These include imaging of cells and living organisms, electrochemical detection of viruses and neurotransmitters, and studies in drug transport and induced-release of adenosine triphosphate from erythrocytes.  相似文献   

10.
BackgroundFor more than a decade, water-soluble, eco-friendly, biocompatible, and low-toxicity fluorescent nanomaterials have received considerable attention for their numerous in vivo and in vitro applications in biomedical imaging, disease diagnostics, and environmental monitoring. Owing to their tunable photoluminescence properties, carbon-based luminescent nanomaterials have shown great potential in bioimaging, photocatalysis, and biosensing among other applications.ResultsMarine environments provide excellent resources for the fabrication of these nanomaterials, because many marine organisms contain interesting trigger organic compounds that can be used as precursors. Herein, we synthesize multi-color emissive carbon dots (CDs) with an intrinsic photoluminescence quantum yield of 20.46%. These nanostructures were achieved through the one-step hydrothermal treatment of marine polysaccharide chondroitin sulfate, obtained from shark cartilage, in aqueous solution.ConclusionsWe successfully demonstrate the low toxicity of our marine resource-derived CDs in zebrafish, and provide an initial assessment of their possible use as a bioimaging agent. Notably, the newly synthesized CDs localize in the intestines of zebrafish larvae, thereby indicating their biocompatibility and potential use as in vivo dyes.How to cite: Kim KW, Choi TY, Kwon YM, et al. Simple synthesis of photoluminescent carbon dots from a marine polysaccharide found in shark cartilage. Electron J Biotechnol 2020;47. https://doi.org/10.1016/j.ejbt.2020.07.003.  相似文献   

11.
We report that modular millifluidic networks are simpler, more cost-effective alternatives to traditional microfluidic networks, and they can be rapidly generated and altered to optimize designs. Droplet traffic can also be studied more conveniently and inexpensively at the millimeter scale, as droplets are readily visible to the naked eye. Bifurcated loops, ladder networks, and parking networks were made using only Tygon® tubing and plastic T-junction fittings and visualized using an iPod® camera. As a case study, droplet traffic experiments through a millifluidic bifurcated loop were conducted, and the periodicity of drop spacing at the outlet was mapped over a wide range of inlet drop spacing. We observed periodic, intermittent, and aperiodic behaviors depending on the inlet drop spacing. The experimentally observed periodic behaviors were in good agreement with numerical simulations based on the simple network model. Our experiments further identified three main sources of intermittency between different periodic and/or aperiodic behaviors: (1) simultaneous entering and exiting events, (2) channel defects, and (3) equal or nearly equal hydrodynamic resistances in both sides of the bifurcated loop. In cases of simultaneous events and/or channel defects, the range of input spacings where intermittent behaviors are observed depends on the degree of inherent variation in input spacing. Finally, using a time scale analysis of syringe pump fluctuations and experiment observation times, we find that in most cases, more consistent results can be generated in experiments conducted at the millimeter scale than those conducted at the micrometer scale. Thus, millifluidic networks offer a simple means to probe collective interactions due to drop traffic and optimize network geometry to engineer passive devices for biological and material analysis.  相似文献   

12.
In recent years, there has been a dramatic increase in the use of poly(dimethylsiloxane) (PDMS) devices for cell-based studies. Commonly, the negative tone photoresist, SU8, is used to pattern features onto silicon wafers to create masters (SU8-Si) for PDMS replica molding. However, the complexity in the fabrication process, low feature reproducibility (master-to-master variability), silane toxicity, and short life span of these masters have been deterrents for using SU8-Si masters for the production of cell culture based PDMS microfluidic devices. While other techniques have demonstrated the ability to generate multiple devices from a single master, they often do not match the high feature resolution (∼0.1 μm) and low surface roughness that soft lithography masters offer. In this work, we developed a method to fabricate epoxy-based masters that allows for the replication of features with high fidelity directly from SU8-Si masters via their PDMS replicas. By this method, we show that we could obtain many epoxy based masters with equivalent features to a single SU8-Si master with a low feature variance of 1.54%. Favorable feature transfer resolutions were also obtained by using an appropriate Tg epoxy based system to ensure minimal shrinkage of features ranging in size from ∼100 μm to <10 μm in height. We further show that surface coating epoxy masters with Cr/Au lead to effective demolding and yield PDMS chambers that are suitable for long-term culturing of sensitive primary hippocampal neurons. Finally, we incorporated pillars within the Au-epoxy masters to eliminate the process of punching media reservoirs and thereby reducing substantial artefacts and wastage.  相似文献   

13.
We report on the feasible fabrication of microfluidic devices for ferroelectric polymers'' synthesis in a rapid and stable fashion. Utilizing micro-mixing and flow-focusing in microchannels, poly(vinylidene fluoride-trifluoroethylene) and copper phthalocyanine are uniformly dispersed in one hydrogel particle, which are then demonstrated to immediate and complete on-chip steady polymerization by moderate ultraviolet treatment. The advantage of our droplet-based microfluidic devices is generating versatile particles from simple spheres to disks or rods, and the lengths of particles can be precisely tuned from 30 to 400 μm through adjusting the flow rates of both disperse and oil phases. In addition, this mixed technique allows for the continuous production of dielectric microparticles with controlled dielectric properties between 10 and 160. Such a microfluidic device offers a flexible platform for multiferroic applications.  相似文献   

14.
A combination of a microfluidic device with a light modulation system was developed to detect the oxygen consumption rate (OCR) of a single developing zebrafish embryo via phase-based phosphorescence lifetime detection. The microfluidic device combines two components: an array of glass microwells containing Pt(II) octaethylporphyrin as an oxygen-sensitive luminescent layer and a microfluidic module with pneumatically actuated glass lids above the microwells to controllably seal the microwells of interest. The total basal respiration (OCR, in pmol O2/min/embryo) of a single developing zebrafish embryo inside a sealed microwell has been successfully measured from the blastula stage (3 h post-fertilization, 3 hpf) through the hatching stage (48 hpf). The total basal respiration increased in a linear and reproducible fashion with embryonic age. Sequentially adding pharmacological inhibitors of bioenergetic pathways allows us to perform respiratory measurements of a single zebrafish embryo at key developmental stages and thus monitor changes in mitochondrial function in vivo that are coordinated with embryonic development. We have successfully measured the metabolic profiles of a single developing zebrafish embryo from 3 hpf to 48 hpf inside a microfluidic device. The total basal respiration is partitioned into the non-mitochondrial respiration, mitochondrial respiration, respiration due to adenosine triphosphate (ATP) turnover, and respiration due to proton leak. The changes in these respirations are correlated with zebrafish embryonic development stages. Our proposed platform provides the potential for studying bioenergetic metabolism in a developing organism and for a wide range of biomedical applications that relate mitochondrial physiology and disease.  相似文献   

15.
With the development of 3D printing techniques, the application of it in microfluidic/Lab-on-a-Chip (LoC) fabrication is becoming more and more attractive. However, to achieve a satisfying printing quality of the target devices, researchers usually require quite an amount of work in calibration trials even for high-end 3D printers. To increase the calibration efficiency of the average priced printers and promote the application of 3D printing technology in the microfluidic community, this work has presented a computer vision (CV)-based method for rapid and precise 3D printing calibration with examples on cylindrical hole/post diameters of 0.2–2.4 mm and rectangular hole/post widths of 0.2–1.0 mm by a stereolithography-based 3D printer. Our method is fully automated, which contains five steps and only needs a camera at hand to provide photos for convolutional neural network recognition. The experimental results showed that our CV-based method could provide calibrated dimensions with just one print of the specific calibration ruler to meet user desire. The higher resolution of the photo provides a higher precision in calibration. Subsequently, only one more print for the target device is needed after the calibration process. Overall, this work has provided a quick and precise calibration tool for researchers to apply 3D printing in the fabrication of their microfluidic/LoC devices with average price printers. Besides, with our open source calibration software and calibration ruler design file, researchers can modify the specific setting based on customized needs and conduct calibration on any type of 3D printer.  相似文献   

16.
Hydrogels have several excellent characteristics suitable for biomedical use such as softness, biological inertness and solute permeability. Hence, integrating hydrogels into microfluidic devices is a promising approach for providing additional functions such as biocompatibility and porosity, to microfluidic devices. However, the poor mechanical strength of hydrogels has severely limited device design and fabrication. A tetra-poly(ethylene glycol) (tetra-PEG) hydrogel synthesized recently has high mechanical strength and is expected to overcome such a limitation. In this research, we have comprehensively studied the implementation of tetra-PEG gel into microfluidic device technology. First, the fabrication of tetra-PEG gel/PDMS hybrid microchannels was established by developing a simple and robust bonding technique. Second, some fundamental features of tetra-PEG gel/PDMS hybrid microchannels, particularly fluid flow and mass transfer, were studied. Finally, to demonstrate the unique application of tetra-PEG-gel-integrated microfluidic devices, the generation of patterned chemical modulation with the maximum concentration gradient: 10% per 20 μm in a hydrogel was performed. The techniques developed in this study are expected to provide fundamental and beneficial methods of developing various microfluidic devices for life science and biomedical applications.  相似文献   

17.
A broad range of microfluidic applications, ranging from cell culture to protein crystallization, requires multilevel devices with different heights and feature sizes (from micrometers to millimeters). While state-of-the-art direct-writing techniques have been developed for creating complex three-dimensional shapes, replication molding from a multilevel template is still the preferred method for fast prototyping of microfluidic devices in the laboratory. Here, we report on a "dry and wet hybrid" technique to fabricate multilevel replication molds by combining SU-8 lithography with a dry film resist (Ordyl). We show that the two lithography protocols are chemically compatible with each other. Finally, we demonstrate the hybrid technique in two different microfluidic applications: (1) a neuron culture device with compartmentalization of different elements of a neuron and (2) a two-phase (gas-liquid) global micromixer for fast mixing of a small amount of a viscous liquid into a larger volume of a less viscous liquid.  相似文献   

18.
《Research Policy》2022,51(5):104505
In this study we attempt to shed more light on the relationship between speed of new technology imitation and the sales performance of the imitator compared to the innovator, with a particular focus on the performance outcomes resulting from the rapid imitation of technologies introduced by the market leader. Using data on handset technologies mounted on more than 600 devices introduced to the UK market by 14 mobile phone vendors operating from 1997 to 2008, we study hundreds of imitative actions to test hypotheses on the extent to which an imitator can catch up (i.e., reduce the market share gap) with the market leader by rapidly imitating its innovations. First, we show that gaining advantage by rapidly imitating a technology pioneer is contingent on whether the pioneer is the market leader or a non-leader rival. Second, we find that the risks of rapid imitation of the market leader's technologies are mitigated when industry clockspeed is high, i.e., during a period of fast innovation and imitation cycles in an industry, resulting in rapid variations in product design. Third, we observe that the degree of competitive responsiveness of the technology pioneer when its innovations are imitated represents an important mechanism that can explain why speed of imitation may affect how an imitator can improve its market share gains relative to the pioneer. This paper advances competitive dynamics and imitation as predictive theories of how rapid imitators might catch up with market leaders in technology-intensive industries.  相似文献   

19.
Precise analysis of the aquatic cells and their responses to the toxic chemicals, i.e., water disinfective agents, is of crucial importance due to their role in the ecosystem. We demonstrate the application of the droplets based millifluidic tool for isolating and longtime monitoring of single Paramecium tetraurelia cells using a large number of water-in-oil emulsion droplets. Due to the automated monitoring of the fluorescence signal, the droplets containing cells are distinguished from the empty reservoirs. A viability indicator is used to follow the metabolic dynamic of the cells in every single droplet. Finally, we perform ecotoxicity tests in droplets, exposing the encapsulated paramecia cells to silver nitrate for determination of EC50 levels, and compare the output with the conventional microtiter plate assay.  相似文献   

20.
Photovoltaic technology offers a sustainable solution to the problem of soaring global energy demands. Recently, metal halide perovskite solar cells (PSCs) have attracted worldwide interest because of their high power conversion efficiency of 25.5% and great potential in becoming a disruptive technology in the photovoltaic industry. The transition from research to commercialization requires advancements of scalable deposition methods for both perovskite and charge transporting thin films. Herein, we share our view regarding the current challenges to fabrication of PSCs by printing techniques. We focus particularly on ink technologies, and summarize the strategies for printing uniform, pinhole-free perovskite films with good crystallinity. Moreover, the stability of perovskite solar modules is discussed and analyzed. We believe this review will be advantageous in the area of printable electronic devices.  相似文献   

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