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1. 发明授权

US06897038B2 Method of laser capture microdissection from a sample utilizing short pulse length 有权
标题翻译：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。

2. 发明授权

US06420132B1 Precision laser capture microdissection utilizing short pulse length 有权
标题翻译：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。

3. 发明授权

US08597715B2 Target activated microtransfer 有权
标题翻译：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。

4. 发明申请

US20100190177A1 TARGET ACTIVATED MICROTRANSFER 有权
标题翻译：目标活化微量RNA
公开(公告)号：US20100190177A1
公开(公告)日：2010-07-29
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： C12Q1/68 , B01L3/00 , G01N33/53 , G01N1/28 , G01N33/566
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。

5. 发明授权

US08460744B2 Target activated microtransfer 有权
标题翻译：靶活化微转移
公开(公告)号：US08460744B2
公开(公告)日：2013-06-11
申请号：US12713105
申请日：2010-02-25
申请人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
发明人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
IPC分类号： B05D1/00
CPC分类号： G01N33/543 , G01N33/56966 , G01N2001/284 , Y10T428/2813
摘要： A device for performing target activated transfer that includes a mounting surface for mounting a tissue sample; and a light source positioned to substantially uniformly irradiate both stained and unstained regions of the tissue sample with light energy that activates the reagent to selectively adhere the stained regions to a transfer surface. Also described is an automated system for transferring tissue from a tissue sample to a transfer substrate. The system includes means for holding a tissue section that includes targets specifically stained with an absorptive stain thereby resulting in a stained tissue surface, and a flexible transfer film that includes a lower thermoplastic layer in sufficient thermal contact with the stained tissue surface; an irradiating assembly configured to provide a predetermined uniform light dose to the entire tissue section; and means for applying a constant pressure to the transfer film during irradiation.
摘要翻译：一种用于执行目标激活转移的装置，其包括用于安装组织样本的安装表面; 以及光源，其定位成基本均匀地照射组织样本的染色和未染色区域，所述光能激发试剂以选择性地将染色区域粘附到转移表面。还描述了一种用于将组织从组织样品转移到转移衬底的自动化系统。该系统包括用于保持组织部分的装置，该组织部分包括用吸收性污渍特异性染色的靶，从而导致染色的组织表面;以及柔性转移膜，其包括与染色的组织表面充分热接触的下部热塑层; 照射组件，被配置为向整个组织部分提供预定的均匀光剂量; 以及用于在照射期间向转印膜施加恒定压力的装置。

6. 发明申请

US20100216166A1 TARGET ACTIVATED MICROTRANSFER 有权
标题翻译：目标活化微量RNA
公开(公告)号：US20100216166A1
公开(公告)日：2010-08-26
申请号：US12713105
申请日：2010-02-25
申请人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
发明人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
IPC分类号： G01N33/567
CPC分类号： G01N33/543 , G01N33/56966 , G01N2001/284 , Y10T428/2813
摘要： A device for performing target activated transfer that includes a mounting surface for mounting a tissue sample; and a light source positioned to substantially uniformly irradiate both stained and unstained regions of the tissue sample with light energy that activates the reagent to selectively adhere the stained regions to a transfer surface. Also described is an automated system for transferring tissue from a tissue sample to a transfer substrate. The system includes means for holding a tissue section that includes targets specifically stained with an absorptive stain thereby resulting in a stained tissue surface, and a flexible transfer film that includes a lower thermoplastic layer in sufficient thermal contact with the stained tissue surface; an irradiating assembly configured to provide a predetermined uniform light dose to the entire tissue section; and means for applying a constant pressure to the transfer film during irradiation.
摘要翻译：一种用于执行目标激活转移的装置，其包括用于安装组织样本的安装表面; 以及光源，其定位成基本均匀地照射组织样本的染色和未染色区域，所述光能激发试剂以选择性地将染色区域粘附到转移表面。还描述了一种用于将组织从组织样品转移到转移衬底的自动化系统。该系统包括用于保持组织部分的装置，该组织部分包括用吸收性污渍特异性染色的靶，从而导致染色的组织表面;以及柔性转移膜，其包括与染色的组织表面充分热接触的下部热塑层; 照射组件，被配置为向整个组织部分提供预定的均匀光剂量; 以及在照射期间向转印膜施加恒定压力的装置。

7. 发明授权

US07709047B2 Target activated microtransfer 有权
标题翻译：靶活化微转移
公开(公告)号：US07709047B2
公开(公告)日：2010-05-04
申请号：US10543218
申请日：2003-07-23
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。

8. 发明授权

US07695752B2 Target activated microtransfer 有权
标题翻译：靶活化微转移
公开(公告)号：US07695752B2
公开(公告)日：2010-04-13
申请号：US11202848
申请日：2005-08-12
申请人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
发明人： Robert F. Bonner , Thomas J. Pohida , Michael R. Emmert-Buck , Michael Anthony Tangrea , Rodrigo F. Chuaqui
IPC分类号： B05D1/00
CPC分类号： G01N33/543 , G01N33/56966 , G01N2001/284 , Y10T428/2813
摘要： A device for performing target activated transfer that includes a mounting surface for mounting a tissue sample; and a light source positioned to substantially uniformly irradiate both stained and unstained regions of the tissue sample with light energy that activates the reagent to selectively adhere the stained regions to a transfer surface. Also described is an automated system for transferring tissue from a tissue sample to a transfer substrate. The system includes means for holding a tissue section that includes targets specifically stained with an absorptive stain thereby resulting in a stained tissue surface, and a flexible transfer film that includes a lower thermoplastic layer in sufficient thermal contact with the stained tissue surface; an irradiating assembly configured to provide a predetermined uniform light dose to the entire tissue section; and means for applying a constant pressure to the transfer film during irradiation.
摘要翻译：一种用于执行目标激活转移的装置，其包括用于安装组织样本的安装表面; 以及光源，其定位成基本均匀地照射组织样本的染色和未染色区域，所述光能激发试剂以选择性地将染色区域粘附到转移表面。还描述了一种用于将组织从组织样品转移到转移衬底的自动化系统。该系统包括用于保持组织部分的装置，该组织部分包括用吸收性污渍特异性染色的靶，从而导致染色的组织表面;以及柔性转移膜，其包括与染色的组织表面充分热接触的下部热塑性层; 照射组件，被配置为向整个组织部分提供预定的均匀光剂量; 以及用于在照射期间向转印膜施加恒定压力的装置。

9. 发明授权

US5502386A Pulsed low frequency EPR spectrometer and imager 失效
公开(公告)号：US5502386A
公开(公告)日：1996-03-26
申请号：US340100
申请日：1994-11-15
申请人： John Bourg , James Mitchell , Mark Mirotznik , Bradley Roth , Sankaran Subramanian , Murali Cherukuri , Paul G. Zablocky , Thomas J. Pohida , Paul D. Smith , Walter S. Friauf , Rolf G. Tschudin
发明人： John Bourg , James Mitchell , Mark Mirotznik , Bradley Roth , Sankaran Subramanian , Murali Cherukuri , Paul G. Zablocky , Thomas J. Pohida , Paul D. Smith , Walter S. Friauf , Rolf G. Tschudin
IPC分类号： G01R33/20 , A61B5/055 , G01N24/10 , G01R33/60
CPC分类号： G01R33/60
摘要： An EPR imager and spectrometer includes pulse generating system for generating broadband pulses having an RF carrier frequency that is not highly absorbed by biological samples. The pulse generating system includes up and down chirp convertors for frequency modulating a carrier frequency pulse and compressing the frequency modulated pulse to form a broadband excitation pulse of high energy. Such a machine could form the basis of a clinical imaging device capable of high sensitivity to free radical species in human patients.

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