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1. 发明授权

US07157223B2 Method and apparatus for detecting enzymatic activity using molecules that change electrophoretic mobility 失效
标题翻译：使用改变电泳迁移率的分子检测酶活性的方法和装置
公开(公告)号：US07157223B2
公开(公告)日：2007-01-02
申请号：US09945396
申请日：2001-08-30
申请人： Nancy Allbritton , Christopher Sims , Michael W. Berns , Gavin D. Meredith , Tatiana B. Krasieva , Bruce J. Tromberg , Chao L. Lee
发明人： Nancy Allbritton , Christopher Sims , Michael W. Berns , Gavin D. Meredith , Tatiana B. Krasieva , Bruce J. Tromberg , Chao L. Lee
IPC分类号： C12Q1/00
CPC分类号： C12M41/46 , C12M35/02 , C12M47/06 , C12Q1/00 , C12Q1/485 , G01N27/44721 , G01N27/44743 , G01N33/50 , G01N33/5017
摘要： The activity of intracellular chemical reactions of molecules is measured by the use of fluorescently labeled substrate molecules that undergo a change in electrophoretic mobility upon chemical reaction such as that catalyzed by an enzyme. Specificity is achieved by using labeled substrate molecules that can be acted upon only by specific enzymes. Thus the activity of a specific enzyme or class of enzymes can be determined. Measurements are made with the intracellular presence of such substrate molecules, at some time of interest, typically after exposure of the cell to a stimulus that activates a particular enzymatic pathway. To ensure accuracy, measurements must be made in a timely manner so as to minimize chemical reactions occurring subsequent to the time of interest. Fast controllable laser lysis is used to obtain the contents of a single cell into which reporter substrate molecules have been introduced. The cell contents are then subjected to capillary electrophoresis and enzymatic activity is determined by comparing amounts of substrate molecules to amounts of enzymatically altered substrate molecules which are separated by the electrophoresis and identified by the presence of a fluorescent label.
摘要翻译：通过使用荧光标记的底物分子来测量分子的细胞内化学反应的活性，所述底物分子在诸如由酶催化的化学反应时经历电泳迁移率的变化。通过使用只能被特定酶作用的标记底物分子来实现特异性。因此，可以确定特异性酶或酶类的活性。通常在感兴趣的某个时间通常在将细胞暴露于激活特定酶促途径的刺激之后，通过细胞内存在这样的底物分子进行测量。为了确保准确性，必须及时进行测量，以最大限度地减少感兴趣的时间后发生的化学反应。使用快速可控的激光裂解来获得引入了报告基质分子的单细胞的含量。然后对细胞内容物进行毛细管电泳，并通过将底物分子的量与通过电泳分离并通过荧光标记的存在鉴定的酶促改变的底物分子的量进行比较来确定酶活性。

2. 发明申请

US20110294208A1 METHOD AND DEVICE FOR CELL SELECTION AND COLLECTION IN AN ISOLATED CULTURING ENVIRONMENT 审中-公开
标题翻译：用于分离培养环境中细胞选择和收集的方法和装置
公开(公告)号：US20110294208A1
公开(公告)日：2011-12-01
申请号：US13145917
申请日：2010-02-11
申请人： Nancy Allbritton , Christopher Sims , Wei Xu
发明人： Nancy Allbritton , Christopher Sims , Wei Xu
IPC分类号： C12N5/16 , C12M1/00 , C12N5/071 , C12M1/12 , C12N1/20 , C12N1/14 , C12N1/16 , C12M3/00 , C12N5/075
CPC分类号： C12M25/04 , C12M23/16
摘要： An apparatus for collecting or culturing cells or cell colonies has a common substrate and a plurality of cell carriers releasably connected to the common substrate. The carriers are arranged in the form of an array. The invention employs microcups as the cell carriers. The substrate is preferably free of barriers between the microcups, and in some embodiments the microcups have porous walls. Methods of using the apparatus are also described.
摘要翻译：用于收集或培养细胞或细胞集落的装置具有公共基底和可释放地连接到共同基底的多个细胞载体。载体以阵列的形式排列。本发明采用微型杯作为细胞载体。基底优选在微杯之间不存在阻挡层，并且在一些实施例中，微型杯具有多孔壁。还描述了使用该装置的方法。

3. 发明授权

US07236888B2 Method to measure the activation state of signaling pathways in cells 有权
标题翻译：测量细胞中信号通路活化状态的方法
公开(公告)号：US07236888B2
公开(公告)日：2007-06-26
申请号：US09990413
申请日：2001-11-21
申请人： Nancy Allbritton , Christopher Sims
发明人： Nancy Allbritton , Christopher Sims
IPC分类号： G06F19/00 , C12Q1/25
CPC分类号： G01N15/1475 , B01L3/5027 , G01N1/02 , G01N33/5005 , G01N33/6803
摘要： The activity of multiple proteins in a single living cell, portion of a cell or in a group of cells is simultaneously measured by introducing reporter molecules into the cell(s) or a portion thereof, chemically modifying the reporter(s) by the enzyme of interest, terminating the modification reactions, removing the reporter(s) and modified reporter(s), and determining the amount of enzyme activity present by measuring or comparing the amount of reporter(s) and modified reporter(s) present. By performing a series of experiments at different time points, conditions, and varieties of cell types, a database is developed for molecular cellular mechanisms in health and disease states. By exposing cells to a variety of compounds data for drug development and screening is provided.
摘要翻译：通过将报告分子引入细胞或其一部分，通过将酶的一个或多个酶的化学修饰报告物同时测量多个蛋白质在单个活细胞，细胞部分或一组细胞中的活性终止修饰反应，除去报告基团和修饰的报告物，以及通过测量或比较存在的报告基团和修饰的报告子的量来测定存在的酶活性的量。通过在不同的时间点，条件和细胞类型的品种进行一系列实验，为健康和疾病状态的分子细胞机制开发了数据库。通过将细胞暴露于各种化合物，提供用于药物开发和筛选的数据。

4. 发明申请

US20070238122A1 SYSTEMS AND METHODS FOR EFFICIENT COLLECTION OF SINGLE CELLS AND COLONIES OF CELLS AND FAST GENERATION OF STABLE TRANSFECTANTS 有权
标题翻译：有效收集单细胞和细胞群的系统和方法以及快速生成稳定转录因子
公开(公告)号：US20070238122A1
公开(公告)日：2007-10-11
申请号：US11733053
申请日：2007-04-09
申请人： Nancy Allbritton , Christopher Sims , Yuli Wang , Mark Bachman , Guann-Pyng Li , Eric Stanbridge
发明人： Nancy Allbritton , Christopher Sims , Yuli Wang , Mark Bachman , Guann-Pyng Li , Eric Stanbridge
IPC分类号： C12Q1/68 , C12N1/20
CPC分类号： G01N1/286 , B01L3/5085 , B01L2200/0668 , B01L2300/0829 , C12M23/12 , C12M47/04 , G01N2001/2886
摘要： A plate manufactured to enable samples of cells, micro-organisms, proteins, DNA, biomolecules and other biological media to be positioned at specific locations or sites on the plate for the purpose of performing addressable analyses on the samples. Preferably, some or all of the sites are built from a removable material or as pallets so that a subset of the samples of interest can be readily isolated from the plate for further processing or analysis. The plate can contain structures or chemical treatments that enhance or promote the attachment and/or function of the samples, and that promote or assist in their analyses. Use of the plate advantageously enables the selection and sorting of cells based on dynamic phenomena and the rapid establishment of stable tranfectants.
摘要翻译：制造的板可使细胞，微生物，蛋白质，DNA，生物分子和其他生物培养基的样品定位在板上的特定位置或位点，以便对样品进行可寻址分析。优选地，一些或所有的部位由可移除的材料或托盘构建，使得感兴趣的样品的子集可以容易地与板分离以用于进一步的处理或分析。该板可以包含增强或促进样品的附着和/或功能的结构或化学处理，并且促进或辅助其分析。板的使用有利于基于动态现象和快速建立稳定的转染剂来选择和分选细胞。

5. 发明申请

US20050237480A1 Chemical modifications to polymer surfaces and the application of polymer grafting to biomaterials 审中-公开
公开(公告)号：US20050237480A1
公开(公告)日：2005-10-27
申请号：US10511788
申请日：2003-05-13
申请人： Nancy Allbritton , Christopher Sims , Guann-Pyng Li , Mark Bachman , Shuwen Hu , Xueqin Ren
发明人： Nancy Allbritton , Christopher Sims , Guann-Pyng Li , Mark Bachman , Shuwen Hu , Xueqin Ren
IPC分类号： C08J7/18 , G01N27/447 , G02B1/04 , G02C7/04
CPC分类号： C08J7/18 , B01J2219/00605 , B01J2219/0061 , B01J2219/0063 , B01J2219/00635 , B01J2219/00637 , G01N27/44704 , G01N27/44752 , G02B1/043
摘要： Polymer-based biomaterials are popular due to ease of fabrication and low costs. However, many polymer substrates have undesirable surface properties. The invention provides a procedure to covalently apply a graft polymer to the surface of a polymer substrate by ultraviolet graft polymerization. The graft polymer is formed from monomers such as PEG, AA, monomethoxy acrylate PEG, HEMA, or DMA. Also, mixed monomers may be used to create the graft and the surface properties of the graft may be tailored for different properties, including hydrophobicity, friction coefficient, electroosmotic mobilities and electrophoretic separations. The invention has particular utility in tailoring surface chemistries in ocular lenses and polymer microdevices. I. II. R:—OHAcrylic Acid(AA) —NH2Acrylamide (AM) —N(CH3)2Dimethylacrylamide (DMA) —OCH2CH2OH2-Hydroxyethylacrylate (HEA) —O(CH2CH2O)nCH3PEG monomethyoxylacrylate (PEG)

6. 发明授权

US09068155B2 Array of micromolded structures for sorting adherent cells 有权
标题翻译：用于分类贴壁细胞的微复合结构阵列
公开(公告)号：US09068155B2
公开(公告)日：2015-06-30
申请号：US13513310
申请日：2012-06-01
申请人： Nancy Allbritton , Christopher Sims , Yuli Wang , Pavak Kirit Shah
发明人： Nancy Allbritton , Christopher Sims , Yuli Wang , Pavak Kirit Shah
IPC分类号： C40B50/06 , C40B60/14 , C12M1/32 , C12M1/12
CPC分类号： C12M23/12 , B01J19/0046 , B01J2219/00452 , B01J2219/00743 , C12M23/20 , C12M23/22 , C12M25/04 , C12M25/16 , C12M47/02 , C12M47/04
摘要： An apparatus for collecting or culturing cells or cell colonies includes: a common substrate formed from a flexible resilient polymeric material and having a plurality of wells formed therein; and a plurality of rigid cell carriers releasably connected to said common substrate, with said carriers arranged in the form of an array, and with each of the carriers resiliently received in one of the wells. A method of collecting or culturing cells or cell colonies with such an apparatus is carried out by depositing a liquid media carrying cells on the apparatus so that said cells settle on or adhere to said the carriers; and then (c) releasing at least one selected carrier having said cells thereon by gradual application of release energy to each carrier from the cavity in which it is received (e.g., by pushing with a probe).
摘要翻译：用于收集或培养细胞或细胞集落的装置包括：由柔性弹性聚合物材料形成且在其中形成有多个孔的共同基底; 以及可释放地连接到所述公共衬底的多个刚性电池载体，所述载体以阵列的形式布置，并且每个载体弹性地容纳在其中一个孔中。使用这种装置收集或培养细胞或细胞集落的方法是通过在装置上沉积携带细胞的液体培养基进行的，以使所述细胞沉降或粘附到所述载体上; 然后（c）通过从其被接收的空腔（例如通过用探针推动）逐渐地向每个载体施加释放能量，从而释放至少一个具有所述细胞的选定载体。

7. 发明申请

US20130065795A1 ARRAY OF MICROMOLDED STRUCTURES FOR SORTING ADHERENT CELLS 有权
公开(公告)号：US20130065795A1
公开(公告)日：2013-03-14
申请号：US13513310
申请日：2012-06-01
申请人： Nancy Allbritton , Christopher Sims , Yuli Wang , Pavak Kirit Shah
发明人： Nancy Allbritton , Christopher Sims , Yuli Wang , Pavak Kirit Shah
IPC分类号： C40B50/06 , C40B60/14
CPC分类号： C12M23/12 , B01J19/0046 , B01J2219/00452 , B01J2219/00743 , C12M23/20 , C12M23/22 , C12M25/04 , C12M25/16 , C12M47/02 , C12M47/04
摘要： An apparatus for collecting or culturing cells or cell colonies includes: a common substrate formed from a flexible resilient polymeric material and having a plurality of wells formed therein; and a plurality of rigid cell carriers releasably connected to said common substrate, with said carriers arranged in the form of an array, and with each of the carriers resiliently received in one of the wells. A method of collecting or culturing cells or cell colonies with such an apparatus is carried out by depositing a liquid media carrying cells on the apparatus so that said cells settle on or adhere to said the carriers; and then (c) releasing at least one selected carrier having said cells thereon by gradual application of release energy to each carrier from the cavity in which it is received (e.g., by pushing with a probe).

8. 发明申请

US20110281743A1 SYSTEMS AND METHODS FOR ISOLATING CELLS IN CELL COLONIES IN CULTURE 审中-公开
标题翻译：用于在细胞培养中分离细胞的系统和方法
公开(公告)号：US20110281743A1
公开(公告)日：2011-11-17
申请号：US13130345
申请日：2009-12-10
申请人： Nancy Allbritton , Christopher Sims , Wei Xu , Hamed Shadpour
发明人： Nancy Allbritton , Christopher Sims , Wei Xu , Hamed Shadpour
IPC分类号： C40B30/00 , C12Q1/68 , H01J49/26 , C40B60/12 , H01J37/26 , C12Q1/02 , G01N33/53
CPC分类号： C12Q1/24 , B01L3/5025 , B01L3/5085 , B01L2200/0668 , B01L2300/0609 , B01L2300/0819
摘要： Selecting and propagating a cell colony of interest from among a plurality of cell colonies carried on a common substrate in culture is carried out by: (a) selecting a cell colony of interest from among the plurality of cell colonies; (b) isolating a cell subset from the cell colony of interest; (c) analyzing (for example, by a destructive analysis) the cell subset isolated from the cell colony of interest to confirm the presence or absence of a desired feature therein; and then (d) propagating the cell colony of interest when the desired feature is present in the cell subset. A micropallet apparatus may include: (a) a substrate; (b) a plurality of discrete arrays formed on the substrate, each of the arrays comprising a plurality of releasable pallets, and (c) a plurality of gap forming regions, wherein the gap forming regions surround the pallets and separate the pallets from one another.
摘要翻译：通过以下步骤进行选择和繁殖培养中携带在共同底物上的多个细胞集落中的感兴趣的细胞集落：（a）从多个细胞集落中选择感兴趣的细胞集落; （b）从感兴趣的细胞集落分离细胞亚群; （c）分析（例如，通过破坏性分析）从感兴趣的细胞集落分离的细胞亚群以确认其中存在或不存在所需特征; 然后（d）当所需特征存在于细胞子集中时，传播感兴趣的细胞集落。微孔装置可以包括：（a）基底; （b）形成在所述基板上的多个分立阵列，每个所述阵列包括多个可释放的托盘，以及（c）多个间隙形成区域，其中所述间隙形成区域围绕所述托盘并将所述托盘彼此分离。

9. 发明申请

US20070128716A1 MICRO-BUBBLE PLATE FOR PATTERNING BIOLOGICAL AND NON-BIOLOGICAL MATERIALS 有权
标题翻译：用于绘制生物和非生物材料的微孔板
公开(公告)号：US20070128716A1
公开(公告)日：2007-06-07
申请号：US11539695
申请日：2006-10-09
申请人： Yuli Wang , Mark Bachman , Christopher Sims , Guann-Pyng Li , Nancy Allbritton
发明人： Yuli Wang , Mark Bachman , Christopher Sims , Guann-Pyng Li , Nancy Allbritton
IPC分类号： C12M1/22
CPC分类号： B01L3/5085 , B01L3/5088 , B01L2200/0642 , B01L2200/12 , B01L2300/0819 , C12M25/06 , C12M29/20
摘要： Systems and methods are provided for patterning biological and non-biological material at specific sites on a plate, as well as growing three dimensional structures. Preferred embodiments comprise a plate with regions that will trap gas, usually in the form of bubbles, when the plate is submerged in liquid. Other embodiment of the present invention include a method for placing materials on the plate at pre-determined locations through the use of trapped gas to prevent materials from collecting at unwanted regions. The plate has great utility for plating cells and tissues at specific sites, such as on an array. The disclosed method can also be used to coat the surface of a plate with coatings at specific locations for patterned coating applications and to build up materials to produce three dimensional structures, including micro-mechanical structures—where the structures may be formed from living or non-living material, tissue or non-tissue, organic or inorganic, and the like.
摘要翻译：提供了系统和方法，用于在板上的特定位置图案化生物和非生物材料，以及生长三维结构。优选实施例包括具有区域的板，当板被浸没在液体中时，其具有通常以气泡形式捕获气体的区域。本发明的其它实施方案包括通过使用捕获的气体将材料放置在预定位置处的材料的方法，以防止材料在不需要的区域收集。该板对于在特定位置（例如阵列）上的电镀细胞和组织具有很大的用途。所公开的方法还可以用于在具有特定位置的涂层涂覆板的表面以用于图案化涂布应用，并且建立材料以产生三维结构，包括微机械结构，其中结构可以由活的或非 - 生物材料，组织或非组织，有机或无机物等。

10. 发明授权

US06740497B2 Method and apparatus for detecting cancerous cells using molecules that change electrophoretic mobility 失效
标题翻译：使用改变电泳迁移率的分子检测癌细胞的方法和装置
公开(公告)号：US06740497B2
公开(公告)日：2004-05-25
申请号：US09859650
申请日：2001-05-17
申请人： Nancy Allbritton , Christopher Sims
发明人： Nancy Allbritton , Christopher Sims
IPC分类号： C12Q148
CPC分类号： C12Q1/00 , B01D53/62 , C12Q1/485 , G01N27/44721 , G01N27/44743 , G01N33/50 , G01N33/5017
摘要： The activity of oncogenic intracellular chemical reactions of molecules is measured by the use of fluorescently labeled substrate molecules that undergo a change in electrophoretic mobility upon a chemical reaction such as that catalyzed by an enzyme or kinase. Specificity is achieved by using labeled substrate molecules that can be acted upon only by specific oncogenic enzymes. Thus the activity of an oncogenic enzyme or class of oncogenic enzymes can be determined. Measurements are made with the intracellular presence of such substrate molecules, at some time of interest. To ensure accuracy, measurements must be made in a timely manner so as to minimize chemical reactions occurring subsequent to the time of interest. Fast controllable laser lysis is used to obtain the contents of said cell or cells into which reporter substrate molecules have been introduced. The cell contents are then subjected to capillary electrophoresis and oncogenic enzymatic activity is determined by comparing amounts of unaltered substrate molecules to the amounts of altered substrate molecules which are separated by the electrophoresis and identified by the presence of a fluorescent label.
摘要翻译：通过使用荧光标记的底物分子来测量分子的致癌细胞内化学反应的活性，所述底物分子在诸如由酶或激酶催化的化学反应上经历电泳迁移率的变化。通过使用可以仅由特定致癌酶起作用的标记底物分子来实现特异性。因此，可以确定致癌酶或致癌酶类的活性。在感兴趣的某个时间，用这种底物分子的细胞内存在进行测量。为了确保准确性，必须及时进行测量，以最大限度地减少感兴趣的时间后发生的化学反应。使用快速可控的激光裂解来获得引入了报告基质分子的所述细胞或细胞的内容物。然后对细胞内容物进行毛细管电泳，并且通过将未改变的底物分子的量与通过电泳分离并通过荧光标记的存在鉴定的改变的底物分子的量进行比较来确定致癌酶活性。

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