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    Non-contact laser capture microdissection
    4.
    发明授权
    Non-contact laser capture microdissection 有权
    非接触式激光捕获显微切割

    公开(公告)号：US06743601B1

    公开(公告)日：2004-06-01

    申请号：US09456042

    申请日：1999-12-06

    申请人： Robert F. Bonner ,  Seth R Goldstein ,  Paul D. Smith ,  Thomas Pohida

    发明人： Robert F. Bonner ,  Seth R Goldstein ,  Paul D. Smith ,  Thomas Pohida

    IPC分类号： G01N130

    CPC分类号： G01N1/2813 ,  G01N2001/284 ,  Y10T428/2813 ,  Y10T428/2839 ,  Y10T436/25375

    摘要： An apparatus and process for the micro juxtaposition is set forth where a selectively activatable surface is maintained spaced apart from the tissue sample and juxtaposed to the tissue sample by activation. In the typical case, activation occurs by laser radiation with the material of the activatable surface thermally expanding and bringing about the desired micro juxtaposition. The disclosed micro juxtapositioning can cause locally and microscopically pressure on tissue sample, insertion to the tissue sample, or contact of an activated or prepared surface to the tissue sample.

    摘要翻译： 阐述了用于微并列的装置和方法，其中可选择性激活的表面保持与组织样本间隔开并且通过激活并置到组织样本。 在典型情况下，通过激活辐射发生激活，其中可激活表面的材料热膨胀并产生所需的微并置。 所公开的微并置可能导致组织样本的局部和显微压力，组织样本的插入或活化或制备的表面与组织样品的接触。

    Method of laser capture microdissection from a sample utilizing short pulse length
    5.
    发明授权
    Method of laser capture microdissection from a sample utilizing short pulse length 有权
    使用短脉冲长度从样品中激光捕获显微解剖的方法

    公开(公告)号：US06897038B2

    公开(公告)日：2005-05-24

    申请号：US10118487

    申请日：2002-04-08

    申请人： Robert F. Bonner ,  Seth R. Goldstein ,  Paul D. Smith ,  Thomas J. Pohida

    发明人： Robert F. Bonner ,  Seth R. Goldstein ,  Paul D. Smith ,  Thomas J. Pohida

    IPC分类号： G01N1/28 ,  G01N1/30

    CPC分类号： G01N1/2813 ,  G01N2001/2833 ,  G01N2001/284

    摘要： Laser capture microdissection occurs where the transfer polymer film is placed on a substrate overlying visualized and selected cellular material from a sample for extraction. The transfer polymer film is focally activated (melted) with a pulse brief enough to allow the melted volume to be confined to that polymer directly irradiated. This invention uses brief pulses to reduce the thermal diffusion into surrounding non-irradiated polymer, preventing it from being heated hot enough to melt while providing sufficient heat by direct absorption in the small focal volume directly irradiated by the focused laser beam. This method can be used both in previously disclosed contact LCM, non contact LCM, using either condenser-side (or beam passes through polymer before tissue) or epi-irradiation (or laser passes through tissue before polymer). It can be used in configuration in which laser passes through tissue before polymer with and without an additional rigid substrate. In its preferred configuration it uses the inertial confinement of the surrounding unmelted thermoplastic polymer (and the overlying rigid substrate) to force expansion of the melted polymer into the underlying tissue target. Utilizing the short pulse protocol, the targeted and extracted material can have a diameter equal to or smaller than the exciting beam.

    摘要翻译： 发生激光捕获显微切割，其中将转移聚合物膜放置在覆盖来自样品的可视化和选择的细胞材料的基底上用于提取。 转移聚合物膜以足够短的脉冲进行焦点活化（熔化），以使熔融体积被限制在直接照射的聚合物上。 本发明使用简短的脉冲来减少对周围未被照射的聚合物的热扩散，防止其被热加热至足以融化，同时通过直接吸收由聚焦激光束直接照射的小焦点体积提供足够的热量。 这种方法可以在以前公开的接触LCM，非接触式LCM中使用，使用冷凝器侧（或者束通过组织之前的聚合物）或外照射（或激光通过聚合物之前的组织）。 它可以用于在具有和不具有额外的刚性基底的聚合物之前激光穿过组织的配置。 在其优选的构造中，它使用周围未熔化的热塑性聚合物（和上覆的刚性基材）的惯性约束来强制将熔融的聚合物膨胀到下面的组织靶中。 利用短脉冲协议，目标和提取的材料的直径可以等于或小于激发光束。

    Method utilizing convex geometry for laser capture microdissection
    6.
    发明授权
    Method utilizing convex geometry for laser capture microdissection 失效
    用于激光捕获显微切割的凸几何的方法

    公开(公告)号：US6100051A

    公开(公告)日：2000-08-08

    申请号：US883821

    申请日：1997-06-27

    申请人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    发明人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    IPC分类号： G01N1/28 ,  G02B21/34 ,  C12Q1/08

    CPC分类号： G02B21/34 ,  G01N1/2813 ,  G01N2001/2833 ,  G01N2001/284 ,  Y10T436/25 ,  Y10T436/25125 ,  Y10T436/25375

    摘要： A process of microdissection where a tissue sample is conventionally visualized in a microscope. A selectively activatable convex surface is provided, preferably on the periphery at the distal end of a rod. This selectively activatable convex surface when locally activated, typically with a laser through an optic light path in the microscope, provides the activated region with adhesive properties. The tissue sample has at least one portion, which is to be extracted is identified. This identified portion is contacted with a portion of the selectively activatable convex surface on the periphery of the rod. When the convex surface is locally activated, typically by exposure to laser light in the footprint of the desired portion, an adhesive transfer surface on the selectively activatable convex surface is activated which adheres to the desired cells in the footprint of the desired portion. Thereafter, the adhesive transfer surface is separated from the remainder of the tissue sample while maintaining adhesion with the portion of the sample. Thus the desired portion of the tissue sample is extracted. The disclosed selectively activatable convex surface is preferably utilized to collect desired tissue samples at more than one location on the same slide or from different slides. A rod having a convex surface with the selectively activatable material is set forth as a staple for use with the apparatus and process. Preferred shapes for the convex surface are disclosed as well as a method for coating rods.

    摘要翻译： 显微切割的过程，其中组织样品通常在显微镜中可视化。 优选地，在棒的远端的周边上设置有选择性激活的凸面。 当局部激活时，这种选择性激活的凸表面通常在显微镜中通过光学光路通过激光提供具有粘合性质的激活区域。 组织样本具有至少一部分要被提取的部分。 该识别部分与杆的周边上的可选择性激活的凸面的一部分接触。 当凸表面被局部激活时，通常通过暴露在所需部分的覆盖区内的激光，激活可选择性激活的凸表面上的粘合剂转移表面，该粘合剂转移表面在期望部分的覆盖区中粘附到期望的电池。 此后，粘合剂转移表面与组织样品的其余部分分离，同时保持与样品部分的粘附。 因此，提取组织样本的期望部分。 所公开的可选择激活的凸表面优选用于在相同载片上或不同载玻片上的多于一个位置收集所需的组织样本。 具有具有可选择性活化材料的凸起表面的杆被设置为与装置和工艺一起使用的订书钉。 公开了用于凸表面的优选形状以及用于涂覆棒的方法。

    Convex geometry adhesive film system for laser capture microdissection
    7.
    发明授权
    Convex geometry adhesive film system for laser capture microdissection 失效
    凸形几何胶片系统用于激光捕获显微切割

    公开(公告)号：US06783734B1

    公开(公告)日：2004-08-31

    申请号：US09387810

    申请日：1999-09-01

    申请人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    发明人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    IPC分类号： B01L1100

    CPC分类号： G02B21/34 ,  G01N1/2813 ,  G01N2001/2833 ,  G01N2001/284 ,  Y10T436/25 ,  Y10T436/25125 ,  Y10T436/25375

    摘要： A tissue sample is conventionally visualized in a microscope. A selectively activated convex surface is provided, preferably at the distal end of a rod. This selectively activated convex surface when activated, typically with a laser through an optic light path in the microscope, provides the activated region with adhesive properties. At least one portion of the tissue sample which is to be extracted is identified. This identified portion is contacted with a portion of the selectively activated convex surface on the end of the rod. When the convex surface is activated, typically by exposure to laser light in the footprint of the desired sample, an adhesive transfer surface on the selectively activated convex surface is provided which adheres to the desired cells in the footprint of the desired sample. Thereafter, the adhesive transfer surface is separated from the remainder of the tissue sample while maintaining adhesion with the desired cells. Thus the desired portion of the tissue sample is extracted. The disclosed selectively activated convex surface is preferably utilized to collect desired tissue samples at more than one location on the same slide or from different slides. The collected tissue samples can thereafter be inspected if desired, as collected on the convex surface, and then liberated—as by dissolving the proteins of the samples. This can effectively concentrate rarely occurring cells in order to obtain enough pure material for analysis. A rod having a convex surface with the selectively activated material is set forth as a staple for use with the apparatus and process. Preferred shapes for the convex surface are disclosed as well as a method for coating rods with a resultant rod article.

    摘要翻译： 传统的组织样本在显微镜下可视化。 提供选择性活化的凸表面，优选地在杆的远端处。 当被激活时，这种选择性活化的凸表面通常通过在显微镜中通过光学光路的激光器提供激活区域的粘合性质。 识别要提取的组织样品的至少一部分。 该识别部分与杆的端部上的选择性活化的凸面的一部分接触。 当凸表面被激活时，通常通过暴露于所需样品的覆盖区域中的激光，在选择性活化的凸表面上提供粘合剂转移表面，该粘合剂转移表面在期望样品的覆盖区中粘附到期望的细胞。 此后，粘合剂转移表面与组织样品的其余部分分离，同时保持与所需细胞的粘附。 因此，提取组织样本的期望部分。 所公开的选择性活化的凸表面优选用于在相同载片上或不同载玻片上的多于一个位置处收集所需的组织样品。 此后，如果需要，收集的组织样品可以如在凸表面上收集的那样被检查，然后通过溶解样品的蛋白质来释放。 这可以有效地集中很少发生的细胞，以获得足够的纯物质进行分析。 具有带有选择性活化材料的凸表面的杆被列为与装置和工艺一起使用的订书钉。 公开了用于凸表面的优选形状以及用所述棒制品涂覆棒的方法。

    Mechanical handling systems for laser capture microdissection
    8.
    发明授权
    Mechanical handling systems for laser capture microdissection 失效
    用于激光捕获显微切割的机械处理系统

    公开(公告)号：US06720191B1

    公开(公告)日：2004-04-13

    申请号：US09601559

    申请日：2000-10-30

    申请人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    发明人： Seth R. Goldstein ,  Robert F. Bonner ,  Paul D. Smith ,  John Peterson ,  Thomas Pohida

    IPC分类号： G01N100

    CPC分类号： G01N1/2813 ,  G01N35/00009 ,  G01N2001/282 ,  G01N2001/284 ,  Y10T436/11 ,  Y10T436/25

    摘要： A method and apparatus of gathering by LCM identified cellular material from randorn locations on a tissue sample to designated locations on a transporting substrate enables convenient further processing. A transporting substrate has an identified mapped location for receiving identified cellular material. At least a segment of a selectively activatable coating is placed on the side of the transporting substrate in apposition to the tissue sample at the mapped location. The transporting substrate and sample are relatively moved to place the selectively activated coating at the mapped location in apposition to identified cellular material of the tissue sample which is to be extracted. Thereafter, the selectively activatable coating is activated and impressed or impressed and activated to form an adhesive region on the transporting substrate for adhering to the identified cellular material. Upon removal of the transporting substrate from the tissue sample, identified cellular material adheres to the transporting substrate at the mapped location.

    摘要翻译： 通过LCM聚集的方法和装置将组织样本上的多个位置的细胞材料鉴定到运输基底上的指定位置，便于进一步处理。 传输基板具有用于接收所识别的细胞材料的识别的映射位置。 至少一段可选择性活化的涂层被放置在运送基片的侧面上，以与所述组织样本在所述位置相对应。 运输基质和样品被相对移动以将选择性活化的涂层置于映射位置处，以与待提取的组织样品的鉴定的细胞材料相关联。 此后，可选择性活化的涂层被活化并印刷或印刷并活化，以在传送基材上形成粘合区域，以粘附到所鉴定的多孔材料上。 当从组织样品中移走运送基质时，鉴定的细胞材料在映射位置附着于运输基质。

    Precision laser capture microdissection utilizing short pulse length
    9.
    发明授权
    Precision laser capture microdissection utilizing short pulse length 有权
    使用短脉冲长度的精密激光捕获显微切割

    公开(公告)号：US06420132B1

    公开(公告)日：2002-07-16

    申请号：US09495401

    申请日：2000-01-31

    申请人： Robert F. Bonner ,  Seth R. Goldstein ,  Paul D. Smith ,  Thomas J. Pohida

    发明人： Robert F. Bonner ,  Seth R. Goldstein ,  Paul D. Smith ,  Thomas J. Pohida

    IPC分类号： G01N130

    CPC分类号： G01N1/2813 ,  G01N2001/2833 ,  G01N2001/284

    摘要： Laser capture microdissection occurs where the transfer polymer film is placed on a substrate overlying visualized and selected cellular material from a sample for extraction. The transfer polymer film is focally activated (melted) with a pulse brief enough to allow the melted volume to be confined to that polymer directly irradiated. This invention uses brief pulses to reduce the thermal diffusion into surrounding non-irradiated polymer, preventing it from being heated hot enough to melt while providing sufficient heat by direct absorption in the small focal volume directly irradiated by the focused laser beam. This method can be used both in previously disclosed contact LCM, non contact LCM, using either condenser-side (or beam passes through polymer before tissue) or epi-irradiation (or laser passes through tissue before polymer). It can be used in configuration in which laser passes through tissue before polymer with and without an additional rigid substrate. In its preferred configuration it uses the inertial confinement of the surrounding unmelted thermoplastic polymer (and the overlying rigid substrate) to force expansion of the melted polymer into the underlying tissue target. Utilizing the short pulse protocol, the targeted and extracted material can have a diameter equal to or smaller than the exciting beam.

    摘要翻译： 发生激光捕获显微切割，其中将转移聚合物膜放置在覆盖来自样品的可视化和选择的细胞材料的基底上用于提取。 转移聚合物膜以足够短的脉冲进行焦点活化（熔化），以使熔融体积被限制在直接照射的聚合物上。 本发明使用简短的脉冲来减少对周围未被照射的聚合物的热扩散，防止其被热加热至足以融化，同时通过直接吸收由聚焦激光束直接照射的小焦点体积提供足够的热量。 这种方法可以在以前公开的接触LCM，非接触式LCM中使用，使用冷凝器侧（或者束通过组织之前的聚合物）或外照射（或激光通过聚合物之前的组织）。 它可以用于在具有和不具有额外的刚性基底的聚合物之前激光穿过组织的配置。 在其优选的构造中，它使用周围未熔化的热塑性聚合物（和上覆的刚性基材）的惯性约束来强制将熔融的聚合物膨胀到下面的组织靶中。 利用短脉冲协议，目标和提取的材料的直径可以等于或小于激发光束。

    Target activated microtransfer
    10.
    发明授权
    Target activated microtransfer 有权
    靶活化微转移

    公开(公告)号：US08597715B2

    公开(公告)日：2013-12-03

    申请号：US12753566

    申请日：2010-04-02

    申请人： Michael R. Emmert-Buck ,  Michael Anthony Tangrea ,  Robert F. Bonner ,  Rodrigo Chuaqui ,  Thomas J. Pohida

    发明人： Michael R. Emmert-Buck ,  Michael Anthony Tangrea ,  Robert F. Bonner ,  Rodrigo Chuaqui ,  Thomas J. Pohida

    IPC分类号： B05D1/00

    CPC分类号： G01N33/56966 ,  G01N33/543 ,  Y10T428/2813 ,  Y10T428/2839

    摘要： A method of removing a target from a biological sample which involves placing a transfer surface in contact with the biological sample, and then focally altering the transfer surface to allow selective separation of the target from the biological sample. In disclosed embodiments, the target is a cell or cellular component of a tissue section and the transfer surface is a film that can be focally altered to adhere the target to the transfer surface. Subsequent separation of the film from the tissue section selectively removes the adhered target from the tissue section. The transfer surface is activated from within the target to adhere the target to the transfer surface, for example by heating the target to adhere it to a thermoplastic transfer surface. Such in situ activation can be achieved by exposing the biological sample to an immunoreagent that specifically binds to the target (or a component of the target). The immunoreagent can alter the transfer surface directly (for example with a heat generating enzyme carried by the immunoreagent), or indirectly (for example by changing a characteristic of the target). In some embodiments, the immunoreagent deposits a precipitate in the target that increases its light absorption relative to surrounding tissue, such that the biological specimen can be exposed to light to selectively heat the target. Alternatively, the immunoreagent is an immunofluorescent agent that carries a fluorophore that absorbs light and emits heat.

    摘要翻译： 一种从生物样品中除去靶的方法，其包括将转移表面与生物样品接触，然后焦点改变转移表面以允许靶与生物样品的选择性分离。 在所公开的实施方案中，靶是组织切片的细胞或细胞组分，并且转移表面是可以被焦点改变以将靶附着到转移表面的膜。 随后从组织切片分离膜，从组织切片选择性地去除粘附的靶。 转移表面从靶内活化，以将靶附着到转印表面，例如通过加热靶以将其粘附到热塑性转印表面。 可以通过将生物样品暴露于特异性结合靶（或靶的成分）的免疫反应物来实现这种原位激活。 免疫反应物可以直接（例如用免疫反应物携带的发热酶）或间接地（例如通过改变靶的特征）来改变转移表面。 在一些实施方案中，免疫反应物在靶中沉积沉淀物，其相对于周围组织增加其光吸收，使得生物样品可暴露于光以选择性加热靶标。 或者，免疫反应剂是携带吸收光并发出热量的荧光团的免疫荧光剂。