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1.
Conventional neuronal analysis at the single neuron level usually involves culturing of neurons in vitro and analysis of neuronal activities by electrophysiological or pharmacological methods. However, the extracellular environments of in vitro neuronal analysis cannot mimic the exact surroundings of the neurons. Here, we report a microfluidic worm-chip for in vivo analysis of neuronal activities upon dynamic chemical stimulations. A comb-shaped microvalve was developed to immobilize whole animal for high-resolution imaging of neuronal activities. Using a sequential sample introduction system, multiple chemical stimuli were delivered to an individual Caenorhabditis elegans nose tip based on programmed interface shifting of laminar flows. ASH sensory neuron responses to various stimuli in individual C. elegans were quantitatively evaluated, and mutants were significantly defective in neuronal responses to certain stimulus in comparison to others. Sensory reduction in the magnitude of the response to repetitive chemical stimulation with different durations was also found. Our study explored the possibility of real-time detection of neuronal activities in individual animals upon multiple stimulations.  相似文献   

2.
微流控芯片通道内径一般约10~50 μm.典型哺乳类动物细胞直径一般为8~30 μm,体积为87 fL~4 pL.  相似文献   

3.
Zhang-Run Xu  Cui-Hong Liu  Jin Fang 《Talanta》2010,80(3):1088-1093
A novel microfluidic chip integrating an osmosis-based micro-pump was developed and used for perfusion cell culture. The micro-pump includes two sealed chambers, i.e., the inner osmotic reagent chamber and the outer water chamber, sandwiching a semi-permeable membrane. The water in the outer chamber was forced to flow through the membrane into the inner chamber via osmosis, facilitating continuous flow of fluidic zone in the channel. An average flow rate of 0.33 μL min−1 was obtained within 50 h along with a precision of 4.3% RSD (n = 51) by using a 100 mg mL−1 polyvinylpyrrolidone (PVP) solution as the osmotic driving reagent and a flow passage area of 0.98 cm2 of the semi-permeable membrane. The power-free micro-pump has been demonstrated to be pulse-free offering stable flow rates during long-term operation. The present microfluidic chip has been successfully applied for the perfusion culture of human colorectal carcinoma cell by continuously refreshing the culture medium with the osmotic micro-pump. In addition, in situ cell immunostaining was also performed on the microchip by driving all the reagent zones with the integrated micro-pump.  相似文献   

4.
设计并验证了一种用于细胞三维培养的集成微柱阵列的微流控芯片.芯片由一片聚二甲基硅氧烷(PDMS)沟道片和一片玻璃盖片组成, 在PDMS沟道片上集成了一个由两排微柱阵列围成的细胞培养室和两条用于输送培养基的侧沟道.微柱间距直接影响了芯片的使用性能, 是整个芯片设计的关键.基于数值模拟和实验验证, 本研究对微柱间距进行了优化设计.优化后的微流控芯片可以很好地实现细胞与细胞外基质模拟材料混合液的稳定注入、培养基中营养物质向培养室内的快速扩散和细胞代谢物的及时排出.在芯片上进行了神经干细胞的三维培养, 证明了芯片上构建的细胞体外微环境的稳定性.  相似文献   

5.
微流控技术由于其固有的优势已发展成为细胞分析中一个强有力的工具.本文从微流控芯片上的细胞培养、细胞微环境的模拟和控制、单细胞分析、芯片器官以及微流控芯片与质谱联用技术等方面对微流控技术在细胞分析研究中的应用进展进行了介绍,并对这一技术的发展前景进行了总结和展望,希望能为相关研究的开展提供启发.  相似文献   

6.
A brief review on biochemical kinetics in the twentieth century mainly concerned with enzyme kinetics and cooperative processes is presented. Molecular biology and, in particular, structural biology provided the basis for modeling biological phenomena at the molecular level. Structure was recognized as the ultimate and only level at which biological processes find an explanation that is satisfactory for chemists and physicists. A new epoch in biology was initiated by successful extensions of the molecular approach from individual molecules and reactions to the cellular and organismic level. Starting with sequencing of whole genomes in the 1980s more and more techniques became available that are suitable for upscaling from molecules to cells. A series of research programs was initiated: genomics dealing with sequencing the DNA of whole organisms, proteomics considering all proteins of a cell and their interactions, metabolomics studying all metabolic reactions of a cell or an organism, and functional genomics or systems biology aiming at an exploration of the dynamics of complete biological entities. At the same time computational facilities have experienced an unexpected development in speed of calculations and storing devices. At present computer simulations of whole cells at molecular resolution are within reach. The challenge for the theorist in biology is to develop methods for handling the enormously complex networks of gene regulation and metabolism in such a way that biological questions can be addressed. This goal cannot be achieved by dynamical systems theory alone. What is needed is a joint effort from different mathematical disciplines supported by empirical knowledge and tools from discrete mathematics to informatics. Two sections with selected examples from our own laboratory dealing with structural bioinformatics of RNA and with a dynamical systems approach to gene regulation are added.  相似文献   

7.
The development of highly efficient analytical methods capable of probing biological systems at system level is an important task that is required in order to meet the requirements of the emerging field of systems biology. Optical molecular imaging (OMI) is a very powerful tool for studying the temporal and spatial dynamics of specific biomolecules and their interactions in real time in vivo. In this article, recent advances in OMI are reviewed extensively, such as the development of molecular probes that make imaging brighter, more stable and more informative (e.g., FPs and semiconductor nanocrystals, also referred to as quantum dots), the development of imaging approaches that provide higher resolution and greater tissue penetration, and applications for measuring biological events from molecule to organism level, including gene expression, protein and subcellular compartment localization, protein activation and interaction, and low-mass molecule dynamics. These advances are of great significance in the field of biological science and could also be applied to disease diagnosis and pharmaceutical screening. Further developments in OMI for systems biology are also proposed.  相似文献   

8.
Caenorhabditis elegans, one of the widely studied model organisms, sense external chemical cues and perform relative chemotaxis behaviors through its simple chemosensory neuronal system. To study the mechanism underlying chemosensory behavior, a rapid and reliable method for quantitatively analyzing the worms' behaviors is essential. In this work, we demonstrated a microfluidic approach for investigating chemotaxis responses of worms to chemical gradients. The flow-based microfluidic chip was consisted of circular tree-like microchannels, which was able to generate eight flow streams containing stepwise chemical concentrations without the difference in flow velocity. Worms' upstream swimming into microchannels with various concentrations was monitored for quantitative analysis of the chemotaxis behavior. By using this microfluidic chip, the attractive and repellent responses of C. elegans to NaCl were successfully quantified within several minutes. The results demonstrated the wild type-like repellent responses and severely impaired attractive responses in grk-2 mutant animals with defects in calcium influx. In addition, the chemotaxis analysis of the third stage larvae revealed that its gustatory response was different from that in the adult stage. Thus, our microfluidic method provided a useful platform for studying the chemosensory behaviors of C. elegans and screening of chemosensation-related chemical drugs.  相似文献   

9.
A combined detection system involving simultaneous LIF and contacfless-conductometric measurements at the same place of the microfluidic chip was described. The LIF measurement was designed according to the confocal principle and a moveable contactless-conduetivity detector was used in C^4D. Both measurements were mutually independent and advantageous in analyses of mixtures. Various experimental parameters affecting the response were examined and optimized. The performances were demonstrated by simultaneous detection of Rhodamine B. And the results showed that the combined detection system could be used sensitively and reliably.  相似文献   

10.
Currently, there has been a growing need for developing in vitro models to better reflect organism response to chemotherapy at tissue level. For this reason, a microfluidic platform was developed for mimicking physiological microenvironment of solid tumor with multicellular tumor spheroids (MTS) for anticancer drug screening. Importantly, the power of this system over traditional systems is that it is simple to operate and high integration in a more physiologically relevant context. As a proof of concept, long-term MTS cultures with uniform structure were realized on the microfluidic based platform. The response of doxorubicin and paclitaxel on different types of spheroids were simultaneously performed by in situ Live/Dead fluorescence stain to provide spatial distribution of dead cells as well as cytotoxicity information. In addition, the established platform combined with microplate reader was capable to determine the cytotoxicity of different sized MTS, showing a more powerful tool than cell staining examination at the end-point of assay. The HCT116 spheroids were then lysed on chip followed by signaling transduction pathway analysis. To our knowledge, the on chip drug screening study is the first to address the drug susceptibility testing and the offline detailed drug signaling pathway analysis combination on one system. Thus, this novel microfluidic platform provides a useful tool for drug screening with tumor spheroids, which is crucial for drug discovery and development.  相似文献   

11.
Mixing a small amount of magnetic beads and regents with large volume samples evenly in microcavities of a microfluidic chip is always the key step for the application of microfluidic technology in the field of magnetophoresis analysis. This article proposes a microfluidic chip for DNA extraction by magnetophoresis, which relies on bubble rising to generate turbulence and microvortices of various sizes to mix magnetic beads with samples uniformly. The construction and working principle of the microfluidic chip are introduced. CFD simulations are conducted when magnetic beads and samples are irritated by the generation of gas bubbles with the variation of supply pressures. The whole mixing process in the microfluidic chip is observed through a high-speed camera and a microfluidic system when the gas bubbles are generated continuously. The influence of supply pressure on the mixing characteristics of the microfluidic chip is investigated and discussed with both simulation and experiments. Compared with magnetic mixing, bubble mixing can avoid the magnetic beads gather phenomenon caused by magnetic forces and provide a rapid and high efficient solution to realize mixing small amount of regents in large volume samples in a certain order without complex moving structures and operations in a chip. Two applications of mixing with the proposed microfluidic chip are also carried out and discussed.  相似文献   

12.
A new cross-linked polystyrene-supported thioanisole reagent is reported. This reagent incorporates the flexible JandaJel™ cross-linker and can be treated with methyl trifluoromethanesulfonate to form the corresponding sulfonium salt. This salt can in turn be deprotonated to form a polymer-supported sulfur ylide that is able to react with aldehydes and ketones to form epoxides. The thioanisole reagent can also be oxidized to form an insoluble sulfoxide reagent that is useful in Swern oxidation reactions. In these reactions, the polymer-supported thioanisole-based reagents can be recovered, regenerated and reused.  相似文献   

13.
Microcalorimetry and measurement of culture turbidity using a Bioscreen C Analyzer System were applied to study the toxic effect of phenanthrene on Cunninghamella elegans IM 1785/21Gp spore germination. The results of C. elegans spore incubation in Bioscreen C microbiology reader showed the inhibition of spore germination by 70% (with 25 mg l−1 of phenanthrene) and total inhibition of the fungus growth with a higher content of the xenobiotic (50-100 mg l−1). The microcalorimetric technique showed to be useful for the estimation of metabolic activity of C. elegans spores in growth medium up to xenobiotic concentrations of 90 mg l−1. These data corresponded with the microscopic observations. The obtained results showed that the microcalorimetry method could be a valuable supplement in the study on the mechanism of PAHs detoxification by fungi.  相似文献   

14.
Five new oxindole alkaloids, gelegamines A-E (1-5), were isolated from the roots of Gelsemium elegans. Their structures were extensively elucidated on the basis of spectroscopic analysis. Among them, the epoxy ring (C-19/C-20) of gelegamine A (1) was assigned as α-orientation by ROESY experiment and DFT method at B3LYP/6-31g(d) level, and gelegamine B (2) is the first humantenine-type alkaloid with 19-(E) ethylidene configuration. The absolute configurations of gelegamines A-E (1-5) were established on biosynthetic consideration coupled with CD experiments.  相似文献   

15.
In the course of automated screening for small-molecule agonists to peroxisome proliferator-activated receptor-γ (PPAR-γ), 10 new linear triterpenes 1-10 have been isolated from the bark of three neocaledonian Cupaniopsis species, SAPINDACEAE. The structures were elucidated by extensive mono- and bi-dimensional spectroscopy and mass spectrometry.  相似文献   

16.
Communication among microorganisms is mediated by secretion and detection of microbial signaling molecules such as quorum-sensing pheromones and microbial hormones. The molecules elicit the regulation of important genes necessary for microbial survival and often play important roles in interspecies or even inter-kingdom communication. Recent progress in the study of the signaling molecules has enabled us to eavesdrop on microbial conversations to gain insight on their intercellular communication system. This review summarizes the recent advances in the chemistry and chemical biology of these important microbial signaling molecules: acyl-homoserine lactones (AHLs), AI-2, CAI-1 related α-hydroxy ketones (AHKs), ComX pheromones, diffusible signal factors (DSFs), diffusible extracellular factor (DF), and Phytophthora mating hormones.  相似文献   

17.
The new method presented in this article achieved the goal of capturing Salmonella typhimurium via immunoreaction and rapid in situ detection of the CdSe/ZnS quantum dots (QDs) labeled S. typhimurium by self-assembly light-emitting diode-induced fluorescence detection (LIF) microsystem on a specially designed multichannel microfluidic chip. CdSe/ZnS QDs were used as fluorescent markers improving detection sensitivity. The microfluidic chip developed in this study was composed of 12 sample channels, 3 mixing zones, and 6 immune reaction zones, which also acted as fluorescence detection zones. QDs–IgG–primary antibody complexes were generated by mixing CdSe/ZnS QDs conjugated secondary antibody (QDs–IgG) and S. typhimurium antibody (primary antibody) in mixing zones. Then, the complexes went into immune reaction zones to label previously captured S. typhimurium in the sandwich mode. The capture rate of S. typhimurium in each detection zone was up to 70%. The enriched QDs-labeled S. typhimurium was detected using a self-assembly LIF microsystem. A good linear relationship was obtained in the range from 3.7 × 10 to 3.7 × 105 cfu mL−1 using the equation I = 0.1739 log (C) − 0.1889 with R2 = 0.9907, and the detection limit was down to 37 cfu mL−1. The proposed method of online immunolabeling with QDs for in situ fluorescence detection on the designed multichannel microfluidic chip had been successfully used to detect S. typhimurium in pork sample, and it has shown potential advantages in practice.  相似文献   

18.
Characterization of the in vivo behavior of nanomaterials aims to optimize their design, to determine their biological effects, and to validate their application. The characteristics of the model organism Caenorhabditis elegans (C. elegans) advocate this 1 mm long nematode as an ideal living system for the primary screening of engineered nanoparticles in a standard synthetic laboratory. This review describes some practicalities and advantages of working with C. elegans that will be of interest for chemists and materials scientists who would like to enter the “worm” community, anticipates some drawbacks, and offers relevant examples of nanoparticle assessment by using C. elegans.  相似文献   

19.
A small library of peculiar biphenyl and terphenyl-containing spirocyclic triones has been synthesized in parallel by combining the organocatalytic three-component domino Knoevenagel/Diels-Alder sequence to Suzuki coupling. This methodology is fast, general and serves as a platform to gain access to novel chemical tools to probe protein-protein interactions.  相似文献   

20.
Gong FC  Tang LH  Shen GL  Yu RQ 《Talanta》2004,62(4):735-740
A fluoroimmunosensing device which was based on ferulic acid (FA)/horseradish peroxidase system for the detection of Schistosoma japonicum antibody (SjAb) has been developed. To circumvent the difficulty of regeneration of immunocomposite surface, a natural chitosan-epoxy resin matrix was used for the immobilization of SjAg. The surface of the immunocomposite layer reacted was easily regenerated by simple polishing. The renewed surface served as a platform for the competitive immuno-reaction of HRP-SjAb and SjAb with SjAg immobilized at the support body surface and for enzymatic reaction. A novel fluorescent substrate ferulic acid for HRP, which is relatively stable toward H2O2, has been adapted in the proposed fluorometric enzyme immunosensing system. FA can been catalyzed to produce a non-fluorescent species. The amount of HRP-SjAb bound to the aforementioned renewable surface layer, which is related to the content of SjAb in samples could be quantitized by measuring the decrease of fluorescence of FA induced by HRP-SjAb. The chitosan incorporated in matrix is favorable for the amplification of this sensing system due to the electrostatic reaction with FA. The proposed method showed a linear response ranging from 45 to 150 ng ml−1, with an improved detection limit of 45 ng ml−1. The method has been employed to determine SjAb in serum samples.  相似文献   

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