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    Methods for the isolation and analysis of cellular protein content
    4.
    发明授权
    Methods for the isolation and analysis of cellular protein content 有权
    分离和分析细胞蛋白质含量的方法

    公开(公告)号：US06969614B1

    公开(公告)日：2005-11-29

    申请号：US09913667

    申请日：2000-02-16

    申请人： Lance A. Liotta ,  Nicole Simone ,  Michael Emmert-Buck ,  Emmanuel F. Petricoin III

    发明人： Lance A. Liotta ,  Nicole Simone ,  Michael Emmert-Buck ,  Emmanuel F. Petricoin III

    IPC分类号： G01N1/18 ,  G01N1/28 ,  G01N1/30 ,  G01N21/00 ,  G01N21/75 ,  G01N33/53 ,  G01N33/68

    CPC分类号： G01N33/6803 ,  G01N2001/284 ,  Y10T436/25375 ,  Y10T436/255

    摘要： The present invention describes devices and methods for performing protein analysis on laser capture microdissected cells, which permits proteomic analysis on cells of different populations. Particular disclosed examples are analysis of normal versus malignant cells, or a comparison of differential protein expression in cells that are progressing from normal to malignant. The protein content of the microdissected cells may be analyzed using techniques such as immunoassays, 1D and 2D gel electrophoresis characterization, Western blotting, liquid chromatography quadrapole ion trap electrospray (LCQ-MS), Matrix Assisted Laser Desorption Ionization/Time of Flight (MALDI/TOF), and Surface Enhanced Laser Desorption Ionization Spectroscopy (SELDI). In addition to permitting direct comparison of qualitative and quantitative protein content of tumor cells and normal cells from the same tissue sample, the methods also allow for investigation of protein characteristics of tumor cells, such as binding ability and amino acid sequence, and differential expression of proteins in particular cell populations in response to drug treatment. The present methods also provide, through the use of protein fingerprinting, a rapid and reliable way to identify the source tissue of a tumor metastasis.

    摘要翻译： 本发明描述了用于对激光捕获显微切割细胞进行蛋白质分析的装置和方法，其允许对不同群体的细胞进行蛋白质组学分析。 特别公开的实例是正常与恶性细胞的分析，或从正常到恶性进展的细胞中差异蛋白表达的比较。 可以使用免疫测定，1D和2D凝胶电泳表征，Western印迹，液相色谱四极离子阱电喷雾（LCQ-MS），基质辅助激光解吸电离/飞行时间（MALDI / TOF）和表面增强激光解吸电离光谱（SELDI）。 除了允许直接比较来自相同组织样品的肿瘤细胞和正常细胞的定性和定量蛋白质含量之外，该方法还允许研究肿瘤细胞的蛋白质特征，例如结合能力和氨基酸序列，以及差异表达 蛋白质在特定的细胞群体响应药物治疗。 本方法还通过使用蛋白质指纹图谱提供了一种快速可靠的方法来鉴定肿瘤转移的源组织。

    Isolation of cellular material under microscopic visualization
    5.
    发明授权
    Isolation of cellular material under microscopic visualization 失效
    在微观可视化下分离细胞材料

    公开(公告)号：US06204030B1

    公开(公告)日：2001-03-20

    申请号：US09388805

    申请日：1999-09-02

    申请人： Lance A. Liotta ,  Michael E. Buck ,  Rhonda Ann Weiss ,  Zhengping Zhuang ,  Robert F. Bonner

    发明人： Lance A. Liotta ,  Michael E. Buck ,  Rhonda Ann Weiss ,  Zhengping Zhuang ,  Robert F. Bonner

    IPC分类号： C12P1912

    CPC分类号： G02B21/32 ,  C12M33/02 ,  G01N1/04 ,  G01N1/08 ,  G01N1/2806 ,  G01N1/2813 ,  G01N1/286 ,  G01N1/31 ,  G01N15/1468 ,  G01N33/50 ,  G01N2001/028 ,  G01N2001/2833 ,  G01N2001/284 ,  G01N2035/00237 ,  Y10T436/25 ,  Y10T436/25125 ,  Y10T436/25375

    摘要： A method of microdissection which involves: forming an image field of cells of the tissue sample utilizing a microscope, identifying at least one zone of cells of interest from the image field of cells which at least one zone of cells of interest includes different types of cells than adjacent zones of cells, and extracting the at least one zone of cells of interest from the tissue sample. The extraction is achieved by contacting the tissue sample with a transfer surface that can be selectively activated so that regions thereof adhere to the zone of cells of interest to be extracted. The transfer surface includes an activatable adhesive layer which provides chemical or electrostatic adherence to the selected regions of the tissue sample. After the transfer surface is activated the transfer surface and tissue sample are separated. During separation the zone of cells of interest remains adhered to the transfer surface and is thus separated from the tissue sample.

    摘要翻译： 一种显微切割的方法，其包括：使用显微镜形成组织样品的细胞的图像场，从细胞的图像区域鉴定感兴趣的细胞的至少一个区域，所述细胞的至少一个区域包括不同类型的细胞 而不是相邻的细胞区域，并且从组织样品中提取感兴趣的细胞的至少一个区域。 通过使组织样品与可以选择性活化的转移表面接触使得其区域粘附到待提取的感兴趣的细胞区域来实现提取。 转移表面包括可激发的粘合剂层，其向组织样品的选定区域提供化学或静电粘附。 转移表面被激活后，分离转移表面和组织样品。 在分离期间，感兴趣的细胞区域保持粘附到转移表面，并因此与组织样品分离。

    Isolation of cellular material under microscopic visualization
    7.
    发明授权
    Isolation of cellular material under microscopic visualization 失效
    在微观可视化下分离细胞材料

    公开(公告)号：US06569639B2

    公开(公告)日：2003-05-27

    申请号：US09765937

    申请日：2001-01-18

    申请人： Lance A. Liotta ,  Michael E. Buck ,  Rhonda Ann Weiss ,  Zhengping Zhuang ,  Robert F. Bonner

    发明人： Lance A. Liotta ,  Michael E. Buck ,  Rhonda Ann Weiss ,  Zhengping Zhuang ,  Robert F. Bonner

    IPC分类号： G01N130

    CPC分类号： G02B21/32 ,  C12M33/02 ,  G01N1/04 ,  G01N1/08 ,  G01N1/2806 ,  G01N1/2813 ,  G01N1/286 ,  G01N1/31 ,  G01N15/1468 ,  G01N33/50 ,  G01N2001/028 ,  G01N2001/2833 ,  G01N2001/284 ,  G01N2035/00237 ,  Y10T436/25 ,  Y10T436/25125 ,  Y10T436/25375

    摘要： A method of microdissection which involves: forming an image field of cells of the tissue sample utilizing a microscope, identifying at least one zone of cells of interest from the image field of cells which at least one zone of cells of interest includes different types of cells than adjacent zones of cells, and extracting the at least one zone of cells of interest from the tissue sample. The extraction is achieved by contacting the tissue sample with a transfer surface that can be selectively activated so that regions thereof adhere to the zone of cells of interest to be extracted. The transfer surface includes an activatable adhesive layer which provides chemical or electrostatic adherence to the selected regions of the tissue sample. After the transfer surface is activated the transfer surface and tissue sample are separated. During separation the zone of cells of interest remains adhered to the transfer surface and is thus separated from the tissue sample.

    摘要翻译： 一种显微切割的方法，其包括：使用显微镜形成组织样品的细胞的图像场，从细胞的图像区域鉴定感兴趣的细胞的至少一个区域，所述细胞的至少一个区域包括不同类型的细胞 而不是相邻的细胞区域，并且从组织样品中提取感兴趣的细胞的至少一个区域。 通过使组织样品与可以选择性活化的转移表面接触使得其区域粘附到待提取的感兴趣的细胞区域来实现提取。 转移表面包括可激发的粘合剂层，其向组织样品的选定区域提供化学或静电粘附。 转移表面被激活后，分离转移表面和组织样品。 在分离期间，感兴趣的细胞区域保持粘附到转移表面，并因此与组织样品分离。

    Hydrogel nanoparticle based immunoassay
    9.
    发明授权
    Hydrogel nanoparticle based immunoassay 有权
    基于水凝胶纳米颗粒的免疫测定

    公开(公告)号：US09012240B2

    公开(公告)日：2015-04-21

    申请号：US13061507

    申请日：2009-08-26

    申请人： Lance A. Liotta ,  Alessandra Luchini ,  Emanuel F. Petricoin ,  Virginia Espina

    发明人： Lance A. Liotta ,  Alessandra Luchini ,  Emanuel F. Petricoin ,  Virginia Espina

    IPC分类号： G01N33/549 ,  G01N33/53 ,  G01N33/544 ,  G01N33/558 ,  G01N33/52 ,  G01N33/543

    CPC分类号： G01N33/521 ,  G01N33/54333 ,  G01N33/54346

    摘要： An immunoassay device incorporating porous polymeric capture nanoparticles within either the sample collection vessel or pre-impregnated into a porous substratum within fluid flow path of the analytical device is presented. This incorporation of capture particles within the immunoassay device improves sensitivity while removing the requirement for pre-processing of samples prior to loading the immunoassay device. A preferred embodiment is coreshell bait containing capture nanoparticles which perform three functions in one step, in solution: a) molecular size sieving, b) target analyte sequestration and concentration, and c) protection from degradation. The polymeric matrix of the capture particles may be made of co-polymeric materials having a structural monomer and an affinity monomer, the affinity monomer having properties that attract the analyte to the capture particle. This device is useful for point of care diagnostic assays for biomedical applications and as field deployable assays for environmental, pathogen and chemical or biological threat identification.

    摘要翻译： 提供了在分析装置的流体流动路径内将多孔聚合物捕获纳米颗粒结合到样品采集容器中或预浸渍到多孔基质内的免疫测定装置。 这种在免疫测定装置内的捕获颗粒的并入提高了灵敏度，同时在加载免疫测定装置之前消除了样品预处理的要求。 优选的实施方案是含有捕获纳米颗粒的芯壳诱饵，其在一个步骤中在溶液中进行三个功能：a）分子筛分，b）目标分析物螯合和浓缩，以及c）防止降解。 捕获颗粒的聚合物基质可以由具有结构单体和亲和单体的共聚物制成，亲和单体具有将分析物吸引到捕获颗粒的性质。 该设备可用于生物医学应用的护理点诊断分析以及环境，病原体和化学或生物威胁鉴定的现场部署测定。

    Tissue preservation and fixation method
    10.
    发明授权
    Tissue preservation and fixation method 有权
    组织保存和固定方法

    公开(公告)号：US08460859B2

    公开(公告)日：2013-06-11

    申请号：US12447773

    申请日：2007-10-26

    申请人： Virginia A. Espina ,  Lance A. Liotta ,  David Geho

    发明人： Virginia A. Espina ,  Lance A. Liotta ,  David Geho

    IPC分类号： A01N1/00

    CPC分类号： G01N1/30 ,  A01N1/00

    摘要： This invention relates, e.g., to a composition that, at room temperature, when contacted with a sample comprising phosphoproteins, can fix and stabilize cellular phosphoproteins, preserve cellular morphology, and allow the sample to be frozen to generate a cryostat frozen section suitable for molecular analysis. The composition comprises (1) a fixative that is effective to fix the phosphoproteins, and that has a sufficient water content to be soluble for a stabilizer and/or a permeability enhancing agent); (2) a stabilizer, comprising (a) a kinase inhibitor and (b) a phosphatase inhibitor and, optionally, (c) a protease (e.g., proteinase) inhibitor; and (3) a permeability enhancing agent (e.g. PEG). Methods are described for preserving phosphoproteins, using such a composition. Also described are endogenous surrogate markers for monitoring protein degradation, including the loss of posttranslational modifications (such as phosphorylation), e.g. the following removal of a cell or tissue from a subject; and exogenous molecular sentinels (e.g. phosphoproteins attached to magnetic nanoparticles) that allow one to evaluate the processing history of a cellular or tissue population sample.

    摘要翻译： 本发明涉及例如在室温下与包含磷蛋白的样品接触时可以固定和稳定细胞磷酸蛋白的组合物，保留细胞形态，并允许样品冷冻以产生适于分子的低温恒温器冷冻切片 分析。 组合物包含（1）固定磷酸蛋白有效的固定剂，并且其具有足够的水含量可溶于稳定剂和/或渗透性增强剂）; （2）稳定剂，其包含（a）激酶抑制剂和（b）磷酸酶抑制剂和任选的（c）蛋白酶（例如蛋白酶）抑制剂; 和（3）渗透性增强剂（例如PEG）。 描述了使用这种组合物来保存磷蛋白的方法。 还描述了用于监测蛋白质降解的内源替代标记，包括翻译后修饰（例如磷酸化）的丧失，例如， 以下从受试者中除去细胞或组织; 和外源性分子前哨蛋白（例如连接到磁性纳米颗粒的磷酸蛋白），其允许评价细胞或组织群体样品的加工历史。