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公开(公告)号：US20070065835A1
公开(公告)日：2007-03-22
申请号：US11231657
申请日：2005-09-20
申请人： Nader Pourmand , Ronald Davis , Shan Wang
发明人： Nader Pourmand , Ronald Davis , Shan Wang
IPC分类号： C12Q1/68
CPC分类号： C12Q1/68 , C12Q1/6837 , C12Q2565/519 , C12Q2537/1373 , C12Q2525/204
摘要： A method of determining the length of a polynucleotide target is provided. With this method, a target is first hybridized to an array of first probes having different, determined lengths, resulting in the formation of duplexes between the polynucleotide target and the first probes. These duplexes have a single stranded section of target if the target is longer than the first probe it is in a duplex with. Next, a second probe having a determined length is hybridized to these duplexes. If the length of the target is greater than the length of the first probe it is displaced during this hybridization step by the process of branch migration. In contrast, if the length of the target is less than or equal to the length of the first probe, it is not displaced. Thus, the length of the polynucleotide target can be determined.
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公开(公告)号：US20060105373A1
公开(公告)日：2006-05-18
申请号：US11271678
申请日：2005-11-10
申请人： Nader Pourmand , Miloslav Karhanek , Ronald Davis
发明人： Nader Pourmand , Miloslav Karhanek , Ronald Davis
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6825 , B82Y15/00 , B82Y30/00 , B82Y40/00 , G01N27/3275 , G01N27/3276
摘要： Methods and apparatus for direct detection of chemical reactions are provided. In a preferred embodiment, electric charge perturbations of the local environment during enzyme-catalyzed reactions are sensed by an electrode system with an immobilized target molecule. The target molecule is preferably DNA. The charge perturbation caused by the polymerase reaction can uniquely identify a DNA sequence. The polymerization process generates local perturbations of charge in the solution near the electrode surface and induces a charge in a polarazible gold electrode. This event is detected as a transient current by a voltage clamp amplifier. Detection of single nucleotides in a sequence can be determined by dispensing individual dNTPs to the electrode solution and detecting the charge perturbations. Alternatively, multiple bases can be determined at the same time using a mix of all dNTPs with subsequent analysis of the resulting signal. The initial enzyme attachment to the DNA molecule can be detected prior to polymerization, with electrode capacitance measurement using the same voltage-clamp amplifier. This technique and device may be adapted to other reaction determinations, such as enzymatic reactions, other electrode configurations, and other amplifying circuits.
摘要翻译： 提供了用于直接检测化学反应的方法和装置。 在优选的实施方案中，酶催化反应期间局部环境的电荷扰动由具有固定化靶分子的电极系统感测。 靶分子优选为DNA。 由聚合酶反应引起的电荷扰动可以唯一地识别DNA序列。 聚合过程在电极表面附近的溶液中产生电荷的局部扰动，并在极化金电极中引起电荷。 该事件由电压钳位放大器检测为瞬态电流。 可以通过将单个dNTP分配到电极溶液并检测电荷扰动来确定序列中单个核苷酸的检测。 或者，可以使用所有dNTP的混合物同时测定多个碱基，随后分析所得信号。 可以在聚合前检测到DNA分子的初始酶附着，使用相同的电压钳位放大器进行电极电容测量。 该技术和装置可以适用于其他反应测定，例如酶反应，其它电极配置和其它放大电路。
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公开(公告)号：US09863939B2
公开(公告)日：2018-01-09
申请号：US12234506
申请日：2008-09-19
IPC分类号： G01N33/00 , G01N33/543 , G01N27/00
CPC分类号： G01N33/54326 , Y10T436/143333
摘要： Methods for analyte detection with magnetic sensors are provided. Aspects of the methods include producing a magnetic sensor device having a magnetically labeled analyte from a sample, such as a serum sample, bound to a surface of a magnetic sensor thereof; and obtaining a signal, e.g., a real-time signal, from the magnetic sensor to determine whether the analyte is present in the sample. Also provided are devices, systems and kits that find use in practicing the methods of the invention. The methods, devices, systems and kits of the invention find use in a variety of different applications, including detection of biomarkers, such as disease markers.
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公开(公告)号：US09598281B2
公开(公告)日：2017-03-21
申请号：US13406269
申请日：2012-02-27
申请人： R. Adam Seger , Paolo Actis , Boaz Vilozny , Nader Pourmand
发明人： R. Adam Seger , Paolo Actis , Boaz Vilozny , Nader Pourmand
IPC分类号： B82Y15/00 , B82Y5/00 , B01L3/00 , G01Q60/44 , G01N33/487
CPC分类号： B82Y5/00 , B01J2219/00371 , B01L3/502715 , B01L3/50273 , B01L2300/0645 , B82Y15/00 , G01N33/48728 , G01Q60/44
摘要： Disclosed herein are methods and systems for controlled ejection of desired material onto surfaces including in single cells using nanopipettes, as well as ejection onto and into cells. Some embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller for depositing a user defined pattern on an arbitrary substrate for the purpose of controlled cell adhesion and growth. Alternate embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller and electronic control of a voltage differential in a bore of the nanopipette electroosmotically injecting material into a cell in a high-throughput manner and with minimal damage to the cell. Yet other embodiments are directed to method and system comprising functionalized nanopipettes combined with scanning ion conductance microscopy for studying molecular interactions and detection of biomolecules inside a single living cell.
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公开(公告)号：US08940142B2
公开(公告)日：2015-01-27
申请号：US12435056
申请日：2009-05-04
IPC分类号： G01N27/327 , G01N27/403 , G01N33/487
CPC分类号： G01N27/42 , G01N27/4035 , G01N33/48707
摘要： Disclosed are methods and devices for biomolecular detection, comprising a nanopipette, exemplified as a hollow inert, non-biological structure with a conical tip opening of nanoscale dimensions, suitable for holding an electrolyte solution which may contain an analyte such as a protein biomolecule to be detected as it is passed through the tip opening. Biomolecules are detected by specific reaction withy peptide ligands chemically immobilized in the vicinity of the tip. Analytes which bind to the ligands cause a detectible change in ionic current. A sensitive detection circuit, using a feedback amplifier circuit, and alternating voltages is further disclosed. Detection of Il-10 at a concentration of 4ng/nl is also disclosed, as is detection of VEGF.
摘要翻译： 公开了用于生物分子检测的方法和装置，其包括纳米管，其示例为具有纳米级尺寸的圆锥形顶端开口的中空惰性非生物结构，适合于保持电解质溶液，所述电解质溶液可以含有诸如蛋白质生物分子的分析物 当它通过尖端开口时检测到。 通过化学固定在尖端附近的肽配体的特异性反应来检测生物分子。 与配体结合的分析物会引起离子电流的可检测变化。 进一步公开了使用反馈放大器电路和交流电压的敏感检测电路。 还公开了浓度为4ng / nl的Il-10的检测，以及VEGF的检测。
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公开(公告)号：US20120225435A1
公开(公告)日：2012-09-06
申请号：US13406269
申请日：2012-02-27
申请人： R. Adam Seger , Paolo Actis , Boaz Vilozny , Nader Pourmand
发明人： R. Adam Seger , Paolo Actis , Boaz Vilozny , Nader Pourmand
CPC分类号： B82Y5/00 , B01J2219/00371 , B01L3/502715 , B01L3/50273 , B01L2300/0645 , B82Y15/00 , G01N33/48728 , G01Q60/44
摘要： Disclosed herein are methods and systems for controlled ejection of desired material onto surfaces including in single cells using nanopipettes, as well as ejection onto and into cells. Some embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller for depositing a user defined pattern on an arbitrary substrate for the purpose of controlled cell adhesion and growth. Alternate embodiments are directed to a method and system comprising nanopipettes combined with an xyz controller and electronic control of a voltage differential in a bore of the nanopipette electroosmotically injecting material into a cell in a high-throughput manner and with minimal damage to the cell. Yet other embodiments are directed to method and system comprising functionalized nanopipettes combined with scanning ion conductance microscopy for studying molecular interactions and detection of biomolecules inside a single living cell.
摘要翻译： 本文公开了用于将所需材料控制性地喷射到包括在使用纳米移液管的单个细胞中的表面的方法和系统，以及喷射到细胞中并进入细胞。 一些实施例涉及包括与xyz控制器组合的纳米移液器的方法和系统，用于在任意基板上沉积用户限定的图案，以便控制细胞粘附和生长。 替代实施例涉及一种方法和系统，其包括与xyz控制器组合的纳米移液器和将纳米片的孔中的电压差的电子控制电子控制以高通量方式以高通量方式电动注入细胞并以最小的细胞损伤进行电子控制。 其他实施方案涉及包括与扫描离子电导显微镜结合的功能化纳米小片剂用于研究单个活细胞内的分子相互作用和生物分子的检测的方法和系统。
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公开(公告)号：US07989396B2
公开(公告)日：2011-08-02
申请号：US11949442
申请日：2007-12-03
申请人： Heng Yu , Nader Pourmand , Shan X. Wang
发明人： Heng Yu , Nader Pourmand , Shan X. Wang
CPC分类号： C40B60/12 , C40B40/08 , C40B40/10 , C40B50/18 , G01N33/54353
摘要： A highly specific and versatile surface chemistry for immobilization of amine-terminated probes is disclosed. A bi-layered polymer thin film serves as the platform for coupling the probes, which are preferably oligonucleotides. The process involves sequentially coating a substrate with polyamine and polyacid anhydride. Hydrolyzed polyacid anhydride groups may be converted to non-hydrolyzed groups at about 100° C. prior to probe attachment. The process of coating the substrate requires no harsh chemical pretreatment of substrates such as RCA or Piranha cleaning. In addition, simple thermal activation of the anhydride groups has a low requirement for storage, leading to a long shelf life of modified surfaces. The disclosed surface chemistry is especially compatible with microfabrication processes, and its effective application to magnetic biosensors is demonstrated.
摘要翻译： 公开了用于固定胺封端的探针的高度特异性和多功能的表面化学。 双层聚合物薄膜用作耦合探针的平台，优选为寡核苷酸。 该方法包括依次用多胺和多酸酐涂覆底物。 在探针附着之前，水解的多酸酐基团可以在约100℃下转化为非水解基团。 涂覆基材的过程不需要对基材进行苛刻的化学预处理，例如RCA或Piranha清洗。 此外，酸酐基团的简单热活化对储存的要求较低，导致改性表面的保质期长。 所公开的表面化学特性与微细加工工艺特别兼容，并且证明了其在磁性生物传感器中的有效应用。
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公开(公告)号：US20090104707A1
公开(公告)日：2009-04-23
申请号：US12234506
申请日：2008-09-19
CPC分类号： G01N33/54326 , Y10T436/143333
摘要： Methods for analyte detection with magnetic sensors are provided. Aspects of the methods include producing a magnetic sensor device having a magnetically labeled analyte from a sample, such as a serum sample, bound to a surface of a magnetic sensor thereof; and obtaining a signal, e.g., a real-time signal, from the magnetic sensor to determine whether the analyte is present in the sample. Also provided are devices, systems and kits that find use in practicing the methods of the invention. The methods, devices, systems and kits of the invention find use in a variety of different applications, including detection of biomarkers, such as disease markers.
摘要翻译： 提供了使用磁传感器进行分析物检测的方法。 所述方法的方面包括制备具有磁性标记的分析物的磁性传感器装置，所述磁性传感器装置与来自其磁性传感器的表面的样品（例如血清样品）结合; 以及从磁传感器获得信号，例如实时信号，以确定分析物是否存在于样品中。 还提供了用于实践本发明的方法的装置，系统和试剂盒。 本发明的方法，装置，系统和试剂盒可用于各种不同的应用，包括检测生物标志物，例如疾病标记物。
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公开(公告)号：US07501253B2
公开(公告)日：2009-03-10
申请号：US11789559
申请日：2007-04-24
申请人： Nader Pourmand , Ronald W. Davis , Shan X. Wang
发明人： Nader Pourmand , Ronald W. Davis , Shan X. Wang
CPC分类号： C12Q1/6837 , C12Q2565/519 , C12Q2537/1373 , C12Q2525/204
摘要： A method of determining the length of a polynucleotide target is provided. With this method, a target is first hybridized to an array of first probes having different, determined lengths, resulting in the formation of duplexes between the polynucleotide target and the first probes. These duplexes have a single stranded section of target if the target is longer than the first probe it is in a duplex with, and a single stranded section of probe if the target is shorter than the first probe it is in a duplex with. Next, a series of probes is hybridized to the duplexes, breaking apart duplexes in which the target and probe have unequal lengths through the process of branch migration. Thus, the target only remains bound in the duplex if the target and probe are of equal lengths. The length of the polynucleotide target can thereby be determined.
摘要翻译： 提供了确定多核苷酸靶标长度的方法。 利用该方法，首先将靶与具有不同的确定长度的第一探针阵列杂交，导致在多核苷酸靶和第一探针之间形成双链体。 如果目标比第一探针长两倍以上，那么这些双链体具有目标的单链段，如果目标短于其双链体中的第一探针，则其具有探针的单链段。 接下来，一系列探针与双链体杂交，分离双链体，其中靶和探针通过分支迁移过程具有不等长度。 因此，如果目标和探针具有相等的长度，则目标仅保持在双工中。 因此可以确定多核苷酸靶的长度。
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公开(公告)号：US08753812B2
公开(公告)日：2014-06-17
申请号：US13434627
申请日：2012-03-29
CPC分类号： C12Q1/6825 , B82Y15/00 , B82Y30/00 , B82Y40/00 , G01N27/3275 , G01N27/3276
摘要： Methods for direct detection of chemical reactions are provided. Electric charge perturbations of the local environment during enzyme-catalyzed reactions are sensed by an electrode system with an immobilized target molecule. The charge perturbation caused by the polymerase reaction can uniquely identify a DNA sequence. The polymerization process generates local perturbations of charge in the solution near the electrode surface and induces a charge in a polarazible gold electrode. This event is detected as a transient current by a voltage clamp amplifier. Detection of single nucleotides in a sequence can be determined by dispensing individual dNTPs to the electrode solution and detecting the charge perturbations. Alternatively, multiple bases can be determined at the same time using a mix of all dNTPs with subsequent analysis of the resulting signal. This technique may be adapted to other reaction determinations, such as enzymatic reactions, other electrode configurations, and other amplifying circuits.
摘要翻译： 提供了直接检测化学反应的方法。 酶催化反应期间局部环境的电荷扰动由具有固定化靶分子的电极系统检测。 由聚合酶反应引起的电荷扰动可以唯一地识别DNA序列。 聚合过程在电极表面附近的溶液中产生电荷的局部扰动，并在极化金电极中引起电荷。 此事件由电压钳位放大器检测为瞬态电流。 可以通过将单个dNTP分配到电极溶液并检测电荷扰动来确定序列中单个核苷酸的检测。 或者，可以使用所有dNTP的混合物同时测定多个碱基，随后分析所得信号。 该技术可以适用于其他反应测定，例如酶反应，其它电极配置和其它放大电路。
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