Application of microfluidic devices in studies of thrombosis and hemostasis

Due to the importance of fluid flow during thrombotic episodes, it is quite appropriate to study clotting and bleeding processes in devices that have well-defined fluid shear environments. Two common devices for applying these defined shear stresses include the cone-and-plate viscometer and parallel-plate flow chamber. While such tools have many salient features, they require large amounts of blood or other protein components. With growth in the area of microfluidics over the last two decades, it has become feasible to miniaturize such flow devices. Such miniaturization not only enables saving of precious samples but also increases the throughput of fluid shear devices, thus enabling the design of combinatorial experiments and making the technique more accessible to the larger scientific community. In addition to simple flows that are common in traditional flow apparatus, more complex geometries that mimic stenosed arteries and the human microvasculature can also be generated. The composition of the microfluidics cell substrate can also be varied for diverse basic science investigations, and clinical investigations that aim to assay either individual patient coagulopathy or response to anti-coagulation treatment. This review summarizes the current state of the art for such microfluidic devices and their applications in the field of thrombosis and hemostasis.     

Influence of different amount of Au on the wetting behavior of PDMS membrane

Polydimethylsiloxane (PDMS) has been widely used as a base material for bio-MEMS/NEMS devices. It is difficult for PDMS to transfer and spread aqueous solution as a kind of highly hydrophobic material. Therefore, surface modification is necessary for PDMS to make it hydrophilic. In this paper, a method of hydrophilization of PDMS surface is proposed. Gold is sputtered to the PDMS substrate by sputter coater in different average thicknesses. Relationship between the average thickness of gold on the PDMS substrate and the contact angle of the surface was studied. It was found that even gold of average thickness less than 1 nm can result in about 25° change of contact angle. AFM is also used to get topographic information of PDMS surface coated with gold. Three cases are classified with different amount of Au: (1) Heterogeneous zone; (2) Transition zone; (3) Film zone. For heterogeneous zone, a simple model about heterogeneous phase wetting is put forward to interpret this phenomenon.     

HS-SPME-GC-MS测定洋甘菊不同部位挥发性成分

目的:分析比较洋甘菊不同部位挥发性成分。方法:采用顶空固相微萃取法(HS-SPME),通过单因素考察依次筛选固相微萃取纤维头、样品量、萃取温度、萃取时间、解吸附时间,得到最佳萃取条件。以该条件萃取洋甘菊不同部位挥发性成分,结合气相色谱-质谱联用法(GC-MS)测定其化学成分,并采用面积归一化法确定各成分的相对含量。结果:以7种主要挥发性成分为指标考察得到了最佳萃取条件,即固相微萃取纤维头为PDMS/DVB型,样品量0.5 g,萃取温度80℃,萃取时间10 min,解吸附时间1 min。从洋甘菊中鉴定出挥发性成分66个,全草、花、根、茎、叶中分别鉴定出挥发性成分40,31,27,29,35个,分别占挥发性成分总峰面积的84.58%,90.27%,82.41%,87.36%,82.10%。结论:洋甘菊不同部位挥发性成分含量(以总峰面积计)由高到低依次为叶全草花茎根;不同部位含共有成分14个,花与全草含共有成分25种,分别占花与全草中已鉴定成分总含量的98%和88%。实验结果为洋甘菊以全草入药提供了化学物质依据。     

A Simple Water‐Based Synthesis of Au Nanoparticle/PDMS Composites for Water Purification and Targeted Drug Release

AuNP/PDMS nanocomposites have been synthesized in the form of gels, foams, and films with distinctive structure and morphology. A simple in situ process in aqueous medium for the formation of such composite materials is described. The nanoparticles are held firmly within the PDMS while still being chemically accessible to substances soluble in PDMS. We demonstrate the utility of this property for water purification applications such as removing aromatic solvents and sulfur‐containing contaminants from water. The contaminants can be freed from the composite with a simple thermal treatment, allowing the material to be reused. We also demonstrate chemically selective uptake and release of a fluorescent dye by the nanocomposite as a drug delivery model system.

     

Autonomous microfluidic multi-channel chip for real-time PCR with integrated liquid handling

We report on a novel, polymer-based, multi-channel device for polymerase chain reaction that combines, for the first time, rapid sample processing in less than 5 min with high throughput at low costs. This is achieved by sample shuttling, during which submicroliter sample plugs (∼100 nl) are oscillated rapidly over three constant-temperature zones by pneumatic actuation with integrated system. The accuracy and the speed of the liquid handling have been significantly increased, while the design of the device can be kept very simple and allows for mass production using conventional low-cost polymer fabrication processes. Massive parallelization can lead to a throughput up to 100 samples in 10 min including the preparation time. The amplification can be optically monitored by means of online fluorescence detection. Successful real-time PCR and the determination of the threshold cycle, C _t, using the developed device were demonstrated with plasmid DNA in a fluorescent real-time format.     

Microfluidic self-assembly of live <Emphasis Type="Italic">Drosophila</Emphasis> embryos for versatile high-throughput analysis of embryonic morphogenesis

A method for assembling Drosophila embryos in a microfluidic device was developed for studies of thermal perturbation of early embryonic development. Environmental perturbation is a complimentary method to injection of membrane-impermeable macromolecules for assaying genetic function and investigating robustness in complex biochemical networks. The development of a high throughput method for perturbing embryos would facilitate the isolation and mapping of signaling pathways. We immobilize Drosophila embryos inside a microfluidic device on minimal potential-energy wells created through surface modification, and thermally perturb these embryos using binary laminar flows of warm and cold solutions. We self-assemble embryos onto oil adhesive pads with an alcohol surfactant carrier fluid (detachment: 0.1 mL/min), and when the surfactant is removed, the embryo-oil adhesion increases to ∼25 mL/min flow rates, which allows for high velocities required for sharp gradients of thermal binary flows. The microfluidic thermal profile was numerically characterized by simulation and experimentally characterized by fluorescence thermometry. The effects of thermal perturbation were observed to induce abnormal morphogenetic movements in live embryos by using time-lapse differential interference contrast (DIC) microscopy.     

A microfluidic platform for sequential ligand labeling and cell binding analysis

Developing biochemical and cell biological assay for screening biomolecules, evaluating their characteristics in biological processes, and determining their pharmacological effects represents a key technology in biomedical research. A PDMS-based integrated microfluidic platform was fabricated and tested for facilitating the labeling of ligand on the nanogram scale and sequential cell binding analysis in a manner that saves both time and reagents. Within this microfluidic platform, ligand labeling, cell immobolization, and optical analysis are performed in a miniaturized, continuous and semi-automated manner. This microfluidic device for ligand labeling and cell analysis is composed of two functional modules: (i) a circular reaction loop for fluorophore-labeling of the ligand and (ii) four parallel-oriented incubation chambers for immobilization of cells, binding of ligand to different cell populations, and optical evaluation of interactions between the labeled ligand and its cell targets. Epidermal growth factor (EGF) as the ligand and different cell lines with various levels of EGF receptor expression have been utilized to test the feasiblity of this microfluidic platform. When compared to studies with traditional Petri dish handling of cells and tissues, or even microwell analyses, experiments with the microfluidic platform described here are much less time consuming, conserve reagents, and are programmable, which makes these platforms a very promising new tool for biological studies.     

Enhancement of Ag85B DNA vaccine immunogenicity against tuberculosis by dissolving microneedles in mice

Tuberculosis (TB) remains a major global public health problem. New immunization methods against TB are urgently needed. Plasmid DNA with a microneedle patch is a potentially attractive strategy to improve the immune effect. A DNA vaccine encoding the secreted protein Ag85B of Mycobacterium tuberculosis was immunized in the skin using microneedles, which can improve protective immunity compared to conventional intramuscular (IM) injection. There is no significant difference between microneedle patch (MNP) and IM immunization when the immunizing dose is low (4.2 μg). However, the results for detecting humoral immunity showed MNP immunization could better provoke an antibody response than IM when the dose is high (12.6 μg). A similar result was observed in cellular immune responses by measuring the cytokines in splenocytes. The effective protection of MNP can also be demonstrated by counting bacteria and analyzing the survival rate. This study indicated that DNA vaccination in the skin using dissolving microneedles may provide a new strategy against TB.     

Stretchable,Multi-Layered Stack Antenna for Smart/Wearable Electronic Applications

The development of microelectronics has been achieved by improving its performance through miniaturization. This was possible through the development of silicon-based semiconductor process technology, but recently, the demand for wearable or flexible devices has increased. These devices are made using various functional elements based on materials that are difficult to utilize with semiconductor devices that contain existing hard silicon-based materials and are bent or flexibly stretched. In this study, wireless antennas suitable for wearable devices were implemented in a stretchable form. It was possible to stably receive a wireless signal, even with a strain of 20% or more, and power light-emitting diodes (LEDs), microheaters, etc. By devising a multi-layered stack antenna without the existing semiconductor process, it was possible to improve the antenna’s reception performance. It is expected that this can be applied in various ways to smart wireless sensors and wearable biomedical devices using the near-field communication (NFC) of smartphones.     

Applications of microfluidics for neuronal studies

Microfabrication processes have changed the technology used in consumer goods, and have now advanced into applications in biology. Microfluidic platforms are microfabricated tools that are gaining popularity for studies of molecular and cellular biology. These platforms can allow precise control of the environment surrounding individual cells and they have been used to study physiologic and pharmacologic responses at the single-cell level. This article reviews microfluidic technology with emphasis on advances that could apply to the study of the nervous system, including architecture for isolation of axons, integrated electrophysiology, patterned physical and chemical substrate cues, and devices for the precisely controlled delivery of possible therapeutic agents such as trophic factors and drugs. The potential of these chips for the study of neurological diseases is also discussed.