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1. 发明申请

US20120135449A1 ITERATIVE STAINING OF BIOLOGICAL SAMPLES 审中-公开
标题翻译：生物样本的迭代染色
公开(公告)号：US20120135449A1
公开(公告)日：2012-05-31
申请号：US13365525
申请日：2012-02-03
申请人： Jun Xie , Robert John Filkins , Fiona Mary Ginty , Michael Christopher Montalto , Anup Sood , Jeffrey Bernard Fortin , Wei-Cheng Tian , Michael John Gerdes
发明人： Jun Xie , Robert John Filkins , Fiona Mary Ginty , Michael Christopher Montalto , Anup Sood , Jeffrey Bernard Fortin , Wei-Cheng Tian , Michael John Gerdes
IPC分类号： G01N21/75 , C12M1/34
CPC分类号： G01N1/312
摘要： Automated methods and devices that facilitate iterative staining of biological samples from imaging applications are provided. The methods include the steps of providing a small volume flow cell containing a biological sample, applying a stain to the biological sample, combining at least two precursor reagents to form an activated destaining agent and wherein the activated destaining agent decomposition rate is greater than or similar to the destaining reaction rate, flowing the destaining agent over the biological sample at a flow rate that is greater than the decomposition rate of the activated destaining agent, and releasing the sample from the flow cell wherein the integrity of the sample is intact. The process of staining, combining and flowing may be iteratively repeated. Also disclosed herein are devices for iterative staining of biological samples comprising a flow cell, in fluid communication with a premixer, wherein the volume capacity of the premixer is smaller than about five times the volume capacity of the flow cell.
摘要翻译：提供了促进成像应用中生物样品的迭代染色的自动化方法和装置。所述方法包括以下步骤：提供含有生物样品的小体积流动池，向生物样品施加染色，将至少两种前体试剂结合以形成活化的脱色剂，并且其中所述活化脱色剂分解速率大于或等于以脱色反应速率使脱色剂以大于活化脱色剂的分解速率的流速流过生物样品，并将样品从流动池中释放出来，其中样品的完整性是完整的。染色，组合和流动的过程可以迭代地重复。本文还公开了用于重复染色生物样品的装置，其包括与预混合器流体连通的流动池，其中预混合器的体积容量小于流动池的体积容量的约五倍。

2. 发明申请

US20140024024A1 METHODS OF DETECTING DNA, RNA AND PROTEIN IN BIOLOGICAL SAMPLES 审中-公开
标题翻译：在生物样品中检测DNA，RNA和蛋白质的方法
公开(公告)号：US20140024024A1
公开(公告)日：2014-01-23
申请号：US13551190
申请日：2012-07-17
申请人： Anup Sood , Michael John Gerdes , Antti Eljas Seppo , Wei Gao , Fiona Mary Ginty , Elizabeth Mary Collins
发明人： Anup Sood , Michael John Gerdes , Antti Eljas Seppo , Wei Gao , Fiona Mary Ginty , Elizabeth Mary Collins
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6841 , C12Q1/6804 , G01N33/53 , C12Q2523/319 , C12Q2543/10 , C12Q2563/107 , C12Q2565/601
摘要： Novel methods of probing multiple targets in a biological sample are provide whereby the targets are DNA, RNA and protein. The method comprises subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe that binds an RNA target, observing a signal, and optionally removing the signal. The method further comprises an antigen retrieval protocol, observing a signal, removing the signal, and optionally applying a protease treatment to access the sample's DNA targets by subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe, observing a signal from the labeled DNA targets, and optionally removing the signal.
摘要翻译：提供探索生物样品中多个靶标的新方法，其目标是DNA，RNA和蛋白质。该方法包括使用结合RNA靶标的标记的核酸探针对样品进行原位杂交反应，观察信号和任选地除去信号。该方法还包括抗原检索方案，观察信号，去除信号，以及任选应用蛋白酶处理以通过使用标记的核酸探针对样品进行原位杂交反应以观察样品的DNA靶标，观察来自标记的DNA靶标，并任选地去除信号。

3. 发明申请

US20110091081A1 METHOD AND SYSTEM FOR ANALYZING THE EXPRESSION OF BIOMARKERS IN CELLS IN SITU IN THEIR TISSUE OF ORIGIN 审中-公开
标题翻译：用于分析生物标志物在其原始组织中的表达的方法和系统
公开(公告)号：US20110091081A1
公开(公告)日：2011-04-21
申请号：US12580651
申请日：2009-10-16
申请人： Brion Daryl Sarachan , Thomas Paul Repoff , Colin Craig McCulloch , Fiona Mary Ginty , Megan Pearl Rothney , Zhengyu Pang
发明人： Brion Daryl Sarachan , Thomas Paul Repoff , Colin Craig McCulloch , Fiona Mary Ginty , Megan Pearl Rothney , Zhengyu Pang
IPC分类号： G06K9/00 , C12Q1/02 , C12Q1/68 , G06F19/00
CPC分类号： C12Q1/6881 , G01N33/5023 , G01N33/5082 , G06T7/11 , G06T7/174 , G06T2207/10056 , G06T2207/20221 , G06T2207/30024
摘要： The present application discloses a technique for obtaining and storing data on expression of multiple biomarkers in individual cells or the compartments of individual cells in a tissue specimen and methods of utilizing that data to create groups, the members of which share some degree of similarity greater than the general population from which the data is drawn, by an analysis of digital images of a portion of the tissue specimen which has been iteratively stained to generate optical signals, typically fluorescent, reflective of the amount of each of the biomarkers examined. The analysis of the images involves a segmentation routine whereby each pixel of the examined images is assigned to an individual cell or a compartment of an individual cell, the intensity of the signal representative of each biomarker is measured for each pixel, a dataset is created in which each cell or compartments of each cell is associated with a signal intensity for each biomarker examined, and the dataset is interrogated with appropriate numerical tools to create groups. It also discloses the display of such groups on images of the tissue examined such that the individual cells belonging to a particular group are marked or indicated on one of the images examined. It further discloses examining the biomarker data for each group for possible association with a biological condition or process in cases in which tissue specimens drawn from different subjects or different portions of the tissue of a subject have been examined. It also discloses using this examination in the diagnoses, prognoses or assessment of the response to therapy of a condition or disease.
摘要翻译：本申请公开了一种用于获得和存储组织样本中个体细胞或个体细胞的多个生物标志物表达的数据的技术，以及利用该数据创建组的方法，其成员具有一定程度的相似性，大于通过对经过迭代染色以产生光信号的组织标本的一部分的数字图像的分析，通常是荧光的，反映了所检查的每个生物标志物的量，绘制数据的一般群体。图像的分析涉及分割程序，其中检查的图像的每个像素被分配给单个单元的单个单元或隔室，针对每个像素测量代表每个生物标志物的信号的强度，数据集被创建在每个细胞的每个细胞或隔室与所检测的每个生物标志物的信号强度相关联，并且用适当的数值工具询问数据集以产生组。它还公开了在所检查的组织的图像上显示这样的组，使得属于特定组的各个细胞在所检查的图像之一上被标记或指示。它还公开了在从受试者的不同受试者或组织的不同部分抽取的组织标本已经被检查的情况下，检查每组的生物标志物数据与生物学条件或过程的可能关联。它还公开了在对病症或疾病的治疗反应的诊断，预后或评估中使用该检查。

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