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公开(公告)号：US08846314B2
公开(公告)日：2014-09-30
申请号：US12716142
申请日：2010-03-02
CPC分类号： G01N27/447 , B01D57/02 , C12N15/101
摘要： A method and system are presented for fast and efficient isolation, purification and quantitation of nucleic acids from complex biological samples using isotachophoresis in microchannels. In an embodiment, a sieving medium may be used to enhance selectivity. In another embodiment, PCR-friendly chemistries are used to purify nucleic acids from complex biological samples and yield nucleic acids ready for further analysis including for PCR. In another embodiment, small RNAs from biological samples are extracted, isolated, preconcentrated and quantitated using on-chip ITP with a high efficiency sieving medium. The invention enables fast concentration and separation (takes 10s to 100s of seconds) of nucleic acids with high selectivity and using lower volumes of reagents (order of 10s of μL to focus less than 1 pg/μL of nucleic acid).
摘要翻译： 提出了一种方法和系统，用于在微通道中使用等速电泳，从复杂生物样品中快速有效地分离，纯化和定量核酸。 在一个实施方案中，可以使用筛分介质来增强选择性。 在另一个实施方案中，使用PCR友好的化学物质从复杂的生物样品中纯化核酸，并产生准备用于进一步分析的核酸，包括用于PCR。 在另一个实施方案中，使用具有高效筛选培养基的片上ITP，提取，分离，预浓缩和定量来自生物样品的小RNA。 本发明使得能够以高选择性快速浓缩和分离（需要10秒至100秒）的核酸，并且使用较低体积的试剂（10μL的浓度以聚焦小于1pg /μL的核酸）。
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2.发明授权On-chip hybridization coupled with ITP based purification for fast sequence specific identification 有权片上杂交与基于ITP的纯化相结合，用于快速序列特异性鉴定公开(公告)号：US08524061B2
公开(公告)日：2013-09-03
申请号：US13373773
申请日：2011-11-29
IPC分类号： G01N27/447 , G01N27/26
CPC分类号： G01N27/44726 , C07K1/26 , C12Q1/68 , C12Q1/6816 , G01N33/6803 , C12Q2525/207 , C12Q2525/301 , C12Q2565/125 , C12Q2565/629
摘要： Isotachophoresis (ITP) can be employed to simultaneously focus the target and ligand of an assay into the same ITP focus zone. The target and ligand can bind to each other in the ITP focus zone, and then the resulting bound complex can be detected (e.g., by fluorescence). The sensitivity of this approach can be greatly increased by the enhanced concentration of both target and ligand that ITP provides in the focus zone. Since ITP can be performed quickly, the resulting assay is both rapid and sensitive. Markers of bacterial urinary tract infections have been experimentally detected at clinically relevant concentrations with this approach. MicroRNA sequences have also been profiled with this approach, which is clinically relevant because MicroRNA is expected to provide useful markers for disease. In one experiment, miR-122 in human kidney and liver was detected and quantified.
摘要翻译： 可以使用等速电泳（ITP）来同时将测定的靶和配体聚焦到相同的ITP聚焦区域中。 靶和配体可以在ITP聚焦区域中彼此结合，然后可以检测所得到的结合复合物（例如通过荧光）。 通过ITP在对焦区域提供的靶和配体的增强浓度，可以大大提高该方法的灵敏度。 由于ITP可以快速进行，因此得到的测定既快速又敏感。 通过这种方法已经在临床相关浓度上实验检测到细菌性尿路感染的标记。 MicroRNA序列也已经用这种方法进行了描述，这是临床上相关的，因为预期MicroRNA可以为疾病提供有用的标记。 在一个实验中，检测并定量人肾脏和肝脏中的miR-122。
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公开(公告)号：US08414754B1
公开(公告)日：2013-04-09
申请号：US11755449
申请日：2007-05-30
IPC分类号： B01D57/02 , G01N27/447 , G01N27/453
CPC分类号： G01N27/44769
摘要： Analysis of samples is facilitated. According to an example embodiment, an electrophoresis approach involves electrophoretically stacking and/or separating a sample or samples. An electrolyte and a mixture of one or more samples with another electrolyte are added to a microchannel or capillary. An electric field is applied to stack (and, in some applications, further separate) the one or more samples. Generally, the electric field and electrolyte are used to facilitate isotachophoretic (ITP) stacking. In some embodiments, a further electric field is applied and used with the electrolyte to facilitate subsequent capillary electrophoresis (CE).
摘要翻译： 样品分析方便。 根据示例性实施方案，电泳方法涉及电泳堆叠和/或分离样品或样品。 将电解质和一种或多种样品与另一种电解质的混合物加入到微通道或毛细管中。 将一个或多个样品施加到堆叠（并且在一些应用中进一步分离）的电场。 通常，电场和电解质用于促进异羟基磷灰石（ITP）堆叠。 在一些实施方案中，施加另外的电场并与电解质一起使用以促进随后的毛细管电泳（CE）。
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4.发明申请On-chip hybridization coupled with ITP based purification for fast sequence specific identification 有权片上杂交与基于ITP的纯化相结合，用于快速序列特异性鉴定公开(公告)号：US20120160689A1
公开(公告)日：2012-06-28
申请号：US13373773
申请日：2011-11-29
IPC分类号： G01N27/447
CPC分类号： G01N27/44726 , C07K1/26 , C12Q1/68 , C12Q1/6816 , G01N33/6803 , C12Q2525/207 , C12Q2525/301 , C12Q2565/125 , C12Q2565/629
摘要： Isotachophoresis (ITP) can be employed to simultaneously focus the target and ligand of an assay into the same ITP focus zone. The target and ligand can bind to each other in the ITP focus zone, and then the resulting bound complex can be detected (e.g., by fluorescence). The sensitivity of this approach can be greatly increased by the enhanced concentration of both target and ligand that ITP provides in the focus zone. Since ITP can be performed quickly, the resulting assay is both rapid and sensitive. Markers of bacterial urinary tract infections have been experimentally detected at clinically relevant concentrations with this approach. MicroRNA sequences have also been profiled with this approach, which is clinically relevant because MicroRNA is expected to provide useful markers for disease. In one experiment, miR-122 in human kidney and liver was detected and quantified.
摘要翻译： 可以使用等速电泳（ITP）来同时将测定的靶和配体聚焦到相同的ITP聚焦区域中。 靶和配体可以在ITP聚焦区域中彼此结合，然后可以检测所得到的结合复合物（例如通过荧光）。 通过ITP在对焦区域提供的靶和配体的增强浓度，可以大大提高该方法的灵敏度。 由于ITP可以快速进行，因此得到的测定既快速又敏感。 通过这种方法已经在临床相关浓度上实验检测到细菌性尿路感染的标记。 MicroRNA序列也已经用这种方法进行了描述，这是临床上相关的，因为预期MicroRNA可以为疾病提供有用的标记。 在一个实验中，检测并定量人肾脏和肝脏中的miR-122。
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公开(公告)号：US20120152746A1
公开(公告)日：2012-06-21
申请号：US13134165
申请日：2011-05-31
IPC分类号： G01N27/447 , B01D57/02
CPC分类号： G01N27/44726
摘要： Indirect detection and/or identification of analytes by ITP can be enhanced by adding a mixture of labeled carrier ampholytes (CAs) to the sample to provide a continuous range of mobility markers. Each analyte can be detected and quantified by corresponding gaps in the CA signal. This approach does not require a priori choice of fluorophores and can be readily applied (without extensive and specific design) to a wide range of analytes. Analyte identification can be expedited by computing a normalized signal integral (NSI) from the CA signals. Empirical calibrations can relate the NSI to effective mobility. Effective mobility results under two or more different pH conditions can be used to determine analyte pKa and fully ionized mobility, which are analyte properties that can facilitate analyte identification.
摘要翻译： 可以通过向样品中加入标记的载体两性电解质（CAs）的混合物来提供ITP的分析物的间接检测和/或鉴定，以提供连续范围的迁移标记。 每个分析物可以通过CA信号中的相应间隙进行检测和量化。 这种方法不需要荧光团的先验选择，并且可以容易地应用于（广泛的和具体的设计）到广泛的分析物。 可以通过从CA信号计算归一化信号积分（NSI）来加速分析物识别。 经验校准可以将NSI与有效移动性相关联。 可以使用两种或多种不同pH条件下的有效迁移率结果来确定分析物pKa和完全电离迁移率，这是分析物的特性，可促进分析物鉴定。
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6.发明授权Electroosmotic micropumps with applications to fluid dispensing and field sampling 失效电渗雾微乳液，适用于流体分配和现场取样公开(公告)号：US07231839B2
公开(公告)日：2007-06-19
申请号：US10638732
申请日：2003-08-11
申请人： David Huber , Shuhuai Yao , Juan G. Santiago
发明人： David Huber , Shuhuai Yao , Juan G. Santiago
CPC分类号： C01B5/00 , B01L3/0241 , B01L2200/143 , B01L2300/069 , B01L2400/0418 , F04B19/006
摘要： Apparatus and methods according to the present invention preferably utilize electroosmotic pumps in fluid handling and field sampling applications. The electroosmotic pumps used in these applications are capable of generating high pressure and flow without moving mechanical parts and the associated generation of unacceptable electrical and acoustic noise, as well as the associated reduction in reliability. Having multiple small flow channels in parallel provides both a high flow rate and high pressure in liquid handling. These electroosmotic pumps are fabricated with materials and structures that improve performance, efficiency, and reduce weight and manufacturing cost relative to presently available micropumps. These electroosmotic pumps also allow for recapture of evolved gases and deposited materials, which may provide for long-term closed-loop operation. Apparatus and methods according to the present invention also allow active regulation of the fluid handling and field sampling devices through electrical control of the flow through the pump. Electroosmotic pumping enables precise dispensing of samples to form genetic, proteomic, or small molecule arrays. A fluid sensor which can be optical, capacitive or other type of electrical sensor is integrated with the electroosmotic pump for measuring liquid levels in liquid drawing and dispensing operations.
摘要翻译： 根据本发明的设备和方法优选地在流体处理和现场取样应用中使用电渗泵。 在这些应用中使用的电渗泵能够在不移动机械部件的情况下产生高压力和流量，并且相关产生不可接受的电气和噪音噪声以及相关联的可靠性降低。 具有多个小流量通道并联提供液体处理中的高流速和高压力。 这些电渗泵由材料和结构制成，相对于目前可获得的微量泵，其材料和结构提高了性能，效率并降低了重量和制造成本。 这些电渗泵还允许重新捕获可能提供长期闭环操作的放出气体和沉积材料。 根据本发明的装置和方法还允许通过对通过泵的流的电控制来主动调节流体处理和现场取样装置。 电渗抽吸可以精确分配样品以形成遗传，蛋白质组或小分子阵列。 可以将光学，电容或其它类型的电传感器的流体传感器与电渗泵一体化，以测量液体抽取和分配操作中的液位。
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公开(公告)号：US07134486B2
公开(公告)日：2006-11-14
申请号：US10272048
申请日：2002-10-16
申请人： Juan G. Santiago , Shulin Zeng
发明人： Juan G. Santiago , Shulin Zeng
IPC分类号： F28D15/00
CPC分类号： B01D67/0062 , B01D19/0031 , B01D61/427 , F04B19/006 , F28D15/00 , F28D15/043 , F28D15/06 , F28F2260/02 , H01L23/473 , H01L2224/48091 , H01L2224/48145 , H01L2224/73257 , H01L2225/06562 , H01L2924/00014 , H01L2924/00012
摘要： Apparatus and methods according to the present invention preferably utilize electroosmotic pumps that are capable of generating high pressure and flow without moving mechanical parts and the associated generation of unacceptable electrical and acoustic noise, as well as the associated reduction in reliability. These electroosmotic pumps are preferably fabricated with materials and structures that improve performance, efficiency, and reduce weight and manufacturing cost relative to presently available micropumps. These electroosmotic pumps also preferably allow for control including venting or recapture of evolved gases and deposited materials, which may provide for long-term closed-loop operation. A gas permeable membrane removes and vents electrolytic gasses generated within the fluid chamber of the electroosmotic pump to ambient. A catalyst can be used to recombine electrolytic gases to form a vapor product that can be vented or condensed into a liquid state. An osmotic membrane provides for re-entry of condensed fluid to the fluid chamber.
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公开(公告)号：US07019972B2
公开(公告)日：2006-03-28
申请号：US10883530
申请日：2004-06-30
IPC分类号： H05K7/20
CPC分类号： G06F1/26 , H01L23/34 , H01L23/473 , H01L23/49827 , H01L2224/05573 , H01L2224/05624 , H01L2224/05644 , H01L2224/05647 , H01L2224/13025 , H01L2224/16 , H01L2924/19105 , H01L2924/00014
摘要： In one aspect, the present invention is a technique of, and a system for conditioning power for a consuming device. In this regard, a power conditioning module, affixed to an integrated circuit device, conditions power to be applied to the integrated circuit device. The power conditioning module includes a semiconductor substrate having a first interface and a second interface wherein the first interface opposes the second interface. The power conditioning module further includes a plurality of interface vias, to provide electrical connection between the first interface and the second interface, and a first set of pads, disposed on the first interface and a second set of pads disposed on the second interface. Each of the pads is connected to a corresponding one of the interface vias on either the first or second interface. The power conditioning module also includes electrical circuitry, disposed within semiconductor substrate, to condition the power to be applied to the integrated circuit device. The electrical circuitry may be disposed on the first interface, the second interface, or both interfaces. Moreover, the electrical circuitry includes at least one voltage regulator and at least one capacitor.
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9.发明申请SIMULTANEOUS EXTRACTION AND SEPARATION OF RNA AND DNA FROM SINGLE CELLS USING ELECTROPHORETIC TECHNIQUES 审中-公开使用电泳技术从单细胞中同时提取和分离RNA和DNA公开(公告)号：US20150191717A1
公开(公告)日：2015-07-09
申请号：US14593925
申请日：2015-01-09
IPC分类号： C12N15/10 , G01N27/447 , C12Q1/68
CPC分类号： A61B1/00009 , A61B1/0005 , A61B1/05 , A61B1/0676 , C12N15/101 , C12Q1/6806 , G01N27/44791 , G06T7/0016 , G06T7/254 , G06T2207/10068 , G06T2207/30032 , C12Q2565/125 , C12Q2565/629
摘要： Devices and methods for preparing RNA and DNA from single cells are disclosed. In particular, the invention relates to a method of simultaneously extracting RNA and DNA from single cells and separating the nucleic acids electrophoretically. An electric field is used to lyse a single target cell, such that the plasma membrane is selectively disrupted without lysing the nuclear membrane. Cytoplasmic RNA is separated from the nucleus by performing isotachophoresis (ITP) in the presence of a sieving matrix that preferentially reduces the mobility of the nucleus. During ITP, the cytoplasmic RNA accumulates at an ITP interface between leading and trailing electrolytes and can later be extracted free of nuclear DNA. The method can be performed in a microfluidic device that fully automates all steps of the process.
摘要翻译： 公开了从单细胞制备RNA和DNA的装置和方法。 特别地，本发明涉及从单个细胞同时提取RNA和DNA并且电泳分离核酸的方法。 电场用于裂解单个靶细胞，使得质膜选择性地破裂而不裂解核膜。 在筛选基质的存在下，通过进行等速电泳（ITP），从细胞质中分离细胞质RNA，其优先降低细胞核的迁移率。 在ITP期间，细胞质RNA在前导电解质和尾部电解质之间的ITP界面处积累，并且可以随后被提取而不含核DNA。 该方法可以在完全自动化该过程的所有步骤的微流体装置中进行。
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10.发明申请公开(公告)号：US20130008789A1
公开(公告)日：2013-01-10
申请号：US13618228
申请日：2012-09-14
申请人： Mostafa Ronaghi , Tarun Khurana , Juan G. Santiago
发明人： Mostafa Ronaghi , Tarun Khurana , Juan G. Santiago
CPC分类号： B01L3/502761 , B01J2219/00317 , B01J2219/00459 , B01J2219/00466 , B01J2219/00596 , B01J2219/00648 , B01J2219/00653 , B01J2219/00702 , B01L2200/0668 , B01L2300/0819 , B01L2400/0415
摘要： Disclosed are a method and apparatus that use an electric field for improved biological assays. The electric field is applied across a device having wells, which receive reactants, which carry a charge. The device thus uses a controllable voltage source between the first and second electrodes, which is controllable to provide a positive charge and a negative charge to a given electrode. By controlled use of the electric field charged species in a fluid in a fluid channel are directed into or out of the well by an electric field between the electrodes. The present method involves the transport of fluids, as in a microfluidic device, and the electric field-induced movement of reactive species according to various assay procedures, such as DNA sequencing, synthesis or the like.
摘要翻译： 公开了使用电场进行改进的生物测定的方法和装置。 电场被施加在具有井的装置上，该装置具有承载电荷的反应物。 因此，该器件在第一和第二电极之间使用可控电压源，其可控制以向给定电极提供正电荷和负电荷。 通过控制使用流体通道中的流体中的电场带电物质通过电极之间的电场被引导进入或离开阱。 本方法涉及如在微流体装置中的流体的运输，以及根据各种测定程序（例如DNA测序，合成等）的反应物种的电场诱导运动。
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