排序方式: 共有7条查询结果,搜索用时 31 毫秒
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Guillaume Mottet Karla Perez-Toralla Ezgi Tulukcuoglu Francois-Clement Bidard Jean-Yves Pierga Irena Draskovic Arturo Londono-Vallejo Stephanie Descroix Laurent Malaquin Jean Louis Viovy 《Biomicrofluidics》2014,8(2)
We present a low cost microfluidic chip integrating 3D micro-chambers for the capture and the
analysis of cells. This device has a simple design and a small footprint. It allows the
implementation of standard biological protocols in a chip format with low volume consumption. The
manufacturing process relies on hot-embossing of cyclo olefin copolymer, allowing the development of
a low cost and robust device. A 3D design of microchannels was used to induce high flow velocity
contrasts in the device and provide a selective immobilization. In narrow distribution channels, the
liquid velocity induces a shear stress that overcomes adhesion forces and prevents cell
immobilization or clogging. In large 3D chambers, the liquid velocity drops down below the threshold
for cell attachment. The devices can be operated in a large range of input pressures and can even be
handled manually using simple syringe or micropipette. Even at high flow injection rates, the 3D
structures protect the captured cell from shear stress. To validate the performances of our device,
we implemented immuno-fluorescence labeling and Fluorescence in Situ Hybridization
(FISH) analysis on cancer cell lines and on a patient pleural effusion sample. FISH is a Food and
Drug Administration approved cancer diagnostic technique that provides quantitative information
about gene and chromosome aberration at the single cell level. It is usually considered as a long
and fastidious test in medical diagnosis. This process can be easily implanted in our platform, and
high resolution fluorescence imaging can be performed with reduced time and computer intensiveness.
These results demonstrate the potential of this chip as a low cost, robust, and versatile tool
adapted to complex and demanding protocols for medical diagnosis. 相似文献
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Paul D Saias L Pedinotti JC Chabert M Magnifico S Pallandre A De Lambert B Houdayer C Brugg B Peyrin JM Viovy JL 《Biomicrofluidics》2011,5(2):24102
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
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Svobodova Z Reza Mohamadi M Jankovicova B Esselmann H Verpillot R Otto M Taverna M Wiltfang J Viovy JL Bilkova Z 《Biomicrofluidics》2012,6(2):24126-2412612
Determination of amyloid β (Aβ) isoforms and in particular the proportion of the Aβ 1-42 isoform in cerebrospinal fluid (CSF) of patients suspected of Alzheimer's disease might help in early diagnosis and treatment of that illness. Due to the low concentration of Aβ peptides in biological fluids, a preconcentration step prior to the detection step is often necessary. This study utilized on-chip immunoprecipitation, known as micro-immunoprecipitation (μIP). The technique uses an immunosorbent (IS) consisting of magnetic beads coated with specific anti-Aβ antibodies organized into an affinity microcolumn by a magnetic field. Our goal was to thoroughly describe the critical steps in developing the IS, such as selecting the proper beads and anti-Aβ antibodies, as well as optimizing the immobilization technique and μIP protocol. The latter includes selecting optimal elution conditions. Furthermore, we demonstrate the efficiency of anti-Aβ IS for μIP and specific capture of 5 Aβ peptides under optimized conditions using various subsequent analytical methods, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), capillary electrophoresis, microchip electrophoresis, and immunoblotting. Synthetic Aβ peptides samples prepared in buffer and spiked in human CSF were analyzed. Finally, on-chip immunoprecipitation of Aβ peptides in human CSF sample was performed. 相似文献
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Reza M. Mohamadi Zuzana Svobodova Zuzana Bilkova Markus Otto Myriam Taverna Stephanie Descroix Jean-Louis Viovy 《Biomicrofluidics》2015,9(5)
We present an integrated microfluidic chip for detection of β-amyloid (Aβ) peptides. Aβ peptides are major biomarkers for the diagnosis of Alzheimer''s disease (AD) in its early stages. This microfluidic device consists of three main parts: (1) An immunocapture microcolumn based on self-assembled magnetic beads coated with antibodies specific to Aβ peptides, (2) a nano-porous membrane made of photopolymerized hydrogel for preconcentration, and (3) a microchip electrophoresis (MCE) channel with fluorescent detection. Sub-milliliter sample volume is either mixed off-chip with antibody coated magnetic beads and injected into the device or is injected into an already self-assembled column of magnetic beads in the microchannel. The captured peptides on the beads are then electrokinetically eluted and re-concentrated onto the nano-membrane in a few nano-liters. By integrating the nano-membrane, total assay time was reduced and also off-chip re-concentration or buffer exchange steps were not needed. Finally, the concentrated peptides in the chip are separated by electrophoresis in a polymer-based matrix. The device was applied to the capture and MCE analysis of differently truncated peptides Aβ (1–37, 1–39, 1–40, and 1–42) and was able to detect as low as 25 ng of synthetic Aβ peptides spiked in undiluted cerebrospinal fluid (CSF). The device was also tested with CSF samples from healthy donors. CSF samples were fluorescently labelled and pre-mixed with the magnetic beads and injected into the device. The results indicated that Aβ1-40, an important biomarker for distinguishing patients with frontotemporal lobe dementia from controls and AD patients, was detectable. Although the sensitivity of this device is not yet enough to detect all Aβ subtypes in CSF, this is the first report on an integrated or semi-integrated device for capturing and analyzing of differently truncated Aβ peptides. The method is less demanding and faster than the conventional Western blotting method currently used for research. 相似文献
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广枣提取液对小鼠免疫功能和运动耐力的影响 总被引:11,自引:0,他引:11
观察了广枣提取液对小鼠免疫功能、运动耐力的影响。结果表明:广枣提取液可明显增加小鼠耐缺氧时间,延长小鼠负重游泳时间,降低运动后小鼠血乳酸水平,显著提高其心肌组织中SOD活力;同时明显增加小鼠脾脏/体重、胸腺/体重比值,增强小鼠对巨噬细胞的吞噬能力.提高小鼠的细胞免疫、体液免疫功能。提示广枣提取液具有显著促进小鼠免疫功能及显著提高小鼠运动耐力的功效。 相似文献
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Zuzana Svobodova Mohamad Reza Mohamadi Barbora Jankovicova Hermann Esselmann Romain Verpillot Markus Otto Myriam Taverna Jens Wiltfang Jean-Louis Viovy Zuzana Bilkova 《Biomicrofluidics》2012,6(2):024126-024126-12
Determination of amyloid β (Aβ) isoforms and in particular the proportion of the Aβ 1-42 isoform in cerebrospinal fluid (CSF) of patients suspected of Alzheimer’s disease might help in early diagnosis and treatment of that illness. Due to the low concentration of Aβ peptides in biological fluids, a preconcentration step prior to the detection step is often necessary. This study utilized on-chip immunoprecipitation, known as micro-immunoprecipitation (μIP). The technique uses an immunosorbent (IS) consisting of magnetic beads coated with specific anti-Aβ antibodies organized into an affinity microcolumn by a magnetic field. Our goal was to thoroughly describe the critical steps in developing the IS, such as selecting the proper beads and anti-Aβ antibodies, as well as optimizing the immobilization technique and μIP protocol. The latter includes selecting optimal elution conditions. Furthermore, we demonstrate the efficiency of anti-Aβ IS for μIP and specific capture of 5 Aβ peptides under optimized conditions using various subsequent analytical methods, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), capillary electrophoresis, microchip electrophoresis, and immunoblotting. Synthetic Aβ peptides samples prepared in buffer and spiked in human CSF were analyzed. Finally, on-chip immunoprecipitation of Aβ peptides in human CSF sample was performed. 相似文献
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