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1. 发明申请

US20140212889A1 METHOD FOR STAINING TISSUE 审中-公开
标题翻译：用于染色组织的方法
公开(公告)号：US20140212889A1
公开(公告)日：2014-07-31
申请号：US14342829
申请日：2012-09-04
申请人： Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
发明人： Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
IPC分类号： G01N33/569
CPC分类号： G01N33/569 , G01N1/30
摘要： It is an object of the present invention to provide a tissue staining method that makes it possible to observe both information on the morphology of a tissue and information on a biological substance such as an antigen molecule to be detected on a single section and in a single view field. The present invention provides a tissue staining method, including carrying out (A) a HE (hematoxylin-eosin) staining, and (B) a histochemical staining, serially on a single tissue section, wherein the histochemical staining is defined as a histochemical technique for detecting a biological substance to be detected in a tissue in a visible manner by use of a binding reaction between the biological substance to be detected and a probe biological substance capable of binding specifically to the biological substance to be detected.
摘要翻译：本发明的目的是提供一种组织染色方法，其可以在单个部分和单个部分上观察关于组织形态的信息和关于待检测的抗原分子的生物物质的信息查看字段。本发明提供一种组织染色方法，包括在单个组织切片上连续进行（A）HE（苏木精 - 伊红）染色和（B）组织化学染色，其中组织化学染色定义为组织化学技术通过使用要检测的生物物质和能够特异性结合待检测的生物物质的探针生物物质之间的结合反应，以可见的方式检测待检测的生物物质。

2. 发明授权

US10551378B2 Tissue staining method 有权
公开(公告)号：US10551378B2
公开(公告)日：2020-02-04
申请号：US14342829
申请日：2012-09-04
申请人： Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
发明人： Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
IPC分类号： G01N33/569
摘要： It is an object of the present invention to provide a tissue staining method that makes it possible to observe both information on the morphology of a tissue and information on a biological substance such as an antigen molecule to be detected on a single section and in a single view field. The present invention provides a tissue staining method, including carrying out (A) a HE (hematoxylin-eosin) staining, and (B) a histochemical staining, serially on a single tissue section, wherein the histochemical staining is defined as a histochemical technique for detecting a biological substance to be detected in a tissue in a visible manner by use of a binding reaction between the biological substance to be detected and a probe biological substance capable of binding specifically to the biological substance to be detected.

3. 发明申请

US20130157287A1 BIOLOGICAL SUBSTANCE DETECTION METHOD 有权
标题翻译：生物物质检测方法
公开(公告)号：US20130157287A1
公开(公告)日：2013-06-20
申请号：US13819453
申请日：2011-08-30
申请人： Kensaku Takanashi , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
发明人： Kensaku Takanashi , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , G01N21/6428 , G01N21/6458 , G01N33/4833 , G01N33/582 , G01N2021/6439 , G01N2021/6441
摘要： A biological substance detection method for detecting a biological substance specifically in a pathological specimen, comprising a step of immunologically staining the pathological specimen using a fluorescent label, a step of staining the pathological specimen with a staining reagent for morphology observation purposes (eosin) to observe the morphology of the pathological specimen, a step of irradiating the stained pathological specimen, with excited light to cause the emission of a fluorescent and detecting the biological substance in the pathological specimen. In the step of immunologically staining the pathological specimen, a special fluorescent particle for which the excitation wavelength appears in a region that is different from the excitation wavelength region of eosin is used as the fluorescent label.
摘要翻译：一种用于特异性检测病理标本中生物物质的生物物质检测方法，包括使用荧光标记对病理标本进行免疫染色的步骤，用形态学观察目的的染色试剂（曙红）染色病理标本的步骤，以观察病理标本的形态，用激发光照射染色的病理标本的步骤，引起荧光发射并检测病理标本中的生物物质。在免疫染色病理标本的步骤中，使用在与曙红的激发波长区域不同的区域中出现激发波长的特殊荧光粒子作为荧光标记。

4. 发明申请

US20130157895A1 TISSUE STAINING METHOD, TISSUE EVALUATION METHOD AND BIOSUBSTANCE DETECTION METHOD 审中-公开
公开(公告)号：US20130157895A1
公开(公告)日：2013-06-20
申请号：US13819574
申请日：2011-03-15
申请人： Takuji Aimiya , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
发明人： Takuji Aimiya , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
IPC分类号： G01N1/30 , G01N33/74
摘要： A tissue staining method which comprises: staining a tissue with a staining reagent wherein a biosubstance recognition site is bonded to particles carrying multiple fluorescent substances accumulated therein; in the stained tissue, counting fluorescent points or measuring fluorescent brightness; and evaluating the expression level of a biosubstance, which matches the biosubstance recognition site, in the aforesaid tissue on the basis of the number of the fluorescent points or fluorescent brightness that was measured.

5. 发明授权

US10809167B2 Tissue staining method with staining agent containing particle holding plural phosphors 有权
公开(公告)号：US10809167B2
公开(公告)日：2020-10-20
申请号：US13819574
申请日：2011-03-15
申请人： Takuji Aimiya , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
发明人： Takuji Aimiya , Hideki Gouda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
IPC分类号： G01N1/30 , B82Y30/00 , B82Y15/00 , G01N33/58 , G01N21/64 , G01N33/74
摘要： A tissue staining method which comprises: staining a tissue with a staining reagent wherein a biosubstance recognition site is bonded to particles carrying multiple fluorescent substances accumulated therein; in the stained tissue, counting fluorescent points or measuring fluorescent brightness; and evaluating the expression level of a biosubstance, which matches the biosubstance recognition site, in the aforesaid tissue on the basis of the number of the fluorescent points or fluorescent brightness that was measured.

6. 发明申请

US20140220598A1 BIOLOGICAL SUBSTANCE DETECTION METHOD 审中-公开
标题翻译：生物物质检测方法
公开(公告)号：US20140220598A1
公开(公告)日：2014-08-07
申请号：US14343411
申请日：2012-09-04
申请人： Kensaku Takanashi , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
发明人： Kensaku Takanashi , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Noriaki Ohuchi , Mika Watanabe
IPC分类号： G01N33/58 , G01N33/574
CPC分类号： G01N33/582 , G01N21/6428 , G01N33/56966 , G01N33/57492 , G01N2021/6439 , G01N2333/70596
摘要： The present invention provides a biological substance detection method for specifically detecting a biological substance from a pathological specimen, by which method, when immunostaining using a fluorescent label and staining for morphological observation using a staining agent for morphological observation are simultaneously performed, the results of fluorescence observation and immunostaining can be assessed properly even if the fluorescent label and/or the staining agent is/are deteriorated by irradiation with an excitation light. The biological substance detection method according to the present invention is characterized in that the brightness retention rate of an immunostained part is in a range of 80% to 120% in relation to the brightness retention rate of apart stained for morphological observation when the fluorescent label used for the immunostaining is observed.
摘要翻译：本发明提供一种用于特异性检测来自病理标本的生物物质的生物物质检测方法，通过该方法，当使用荧光标记物进行免疫染色并同时进行用于形态学观察的染色剂进行形态学观察的染色时，荧光的结果即使通过用激发光照射使荧光标记和/或染色剂劣化，也可以适当地评估观察和免疫染色。根据本发明的生物物质检测方法的特征在于，当使用荧光标记物时，免疫染色部分的亮度保持率相对于形态观察时分离染色的亮度保持率在80％至120％的范围内观察免疫染色。

7. 发明授权

US08969100B2 Method for detecting afferent lymph vessel inflow regions and method for identifying specific cells 有权
标题翻译：检测传入淋巴管流入区域的方法和识别特定细胞的方法
公开(公告)号：US08969100B2
公开(公告)日：2015-03-03
申请号：US13379320
申请日：2010-03-11
申请人： Makoto Hikage , Kohsuke Gonda , Motohiro Takeda , Takashi Kamei , Noriaki Ohuchi , Hideki Gouda , Yasushi Nakano
发明人： Makoto Hikage , Kohsuke Gonda , Motohiro Takeda , Takashi Kamei , Noriaki Ohuchi , Hideki Gouda , Yasushi Nakano
IPC分类号： G01N33/543 , G01N33/574 , G01N21/64 , A61K49/00 , B82Y5/00 , B82Y15/00 , G01N33/58 , A61B5/00
CPC分类号： G01N21/6458 , A61B5/0071 , A61B5/41 , A61B5/418 , A61K49/0017 , A61K49/0067 , B82Y5/00 , B82Y15/00 , G01N33/57492 , G01N33/588 , G01N2201/0221 , Y10T436/13
摘要： Regions where metastatic cancer cells can exist are detected with high accuracy in a sentinel lymph node. Quantum dots are injected into the vicinity of a cancer in a living body, thereby identifying the location of the sentinel lymph node by means of fluorescence. Subsequently, the sentinel lymph node is extracted. With respect to the sentinel lymph node extracted with quantum dots injected, structural analysis is conducted by means of precision fluorescence measurement which uses a confocal fluorescence microscope for monomolecular observation. Specifically, the fluorescence intensity is measured with respect to each of multiple areas in the sentinel lymph nodes, and out of the multiple areas measured, one or more areas are detected as afferent lymph vessel inflow regions in descending order of fluorescence intensity.
摘要翻译：在前哨淋巴结中以高精度检测转移性癌细胞可存在的区域。将量子点注入生物体内的癌症附近，通过荧光鉴定前哨淋巴结的位置。随后，提取前哨淋巴结。对于注射量子点提取的前哨淋巴结，采用精细荧光测量法进行结构分析，采用共焦荧光显微镜进行单分子观察。具体地，针对前哨淋巴结中的多个区域中的每个区域测量荧光强度，并且在测量的多个区域中，以荧光强度的降序将一个或多个区域作为传入淋巴管流入区域检测。

8. 发明申请

US20120107831A1 METHOD FOR DETECTING AFFERENT LYMPH VESSEL INFLOW REGIONS AND METHOD FOR IDENTIFYING SPECIFIC CELLS 有权
标题翻译：用于检测感染性淋巴管流入区域的方法和鉴定特异性细胞的方法
公开(公告)号：US20120107831A1
公开(公告)日：2012-05-03
申请号：US13379320
申请日：2010-03-11
申请人： Makoto Hikage , Kohsuke Gonda , Motohiro Takeda , Takashi Kamei , Noriaki Ohuchi , Hideki Gouda , Yasushi Nakano
发明人： Makoto Hikage , Kohsuke Gonda , Motohiro Takeda , Takashi Kamei , Noriaki Ohuchi , Hideki Gouda , Yasushi Nakano
IPC分类号： G01N21/64 , B82Y5/00
CPC分类号： G01N21/6458 , A61B5/0071 , A61B5/41 , A61B5/418 , A61K49/0017 , A61K49/0067 , B82Y5/00 , B82Y15/00 , G01N33/57492 , G01N33/588 , G01N2201/0221 , Y10T436/13
摘要： Regions where metastatic cancer cells can exist are detected with high accuracy in a sentinel lymph node. Quantum dots are injected into the vicinity of a cancer in a living body, thereby identifying the location of the sentinel lymph node by means of fluorescence. Subsequently, the sentinel lymph node is extracted. With respect to the sentinel lymph node extracted with quantum dots injected, structural analysis is conducted by means of precision fluorescence measurement which uses a confocal fluorescence microscope for monomolecular observation. Specifically, the fluorescence intensity is measured with respect to each of multiple areas in the sentinel lymph nodes, and out of the multiple areas measured, one or more areas are detected as afferent lymph vessel inflow regions in descending order of fluorescence intensity.
摘要翻译：在前哨淋巴结中以高精度检测转移性癌细胞可存在的区域。将量子点注入生物体内的癌症附近，通过荧光鉴定前哨淋巴结的位置。随后，提取前哨淋巴结。对于注射量子点提取的前哨淋巴结，采用精细荧光测量法进行结构分析，采用共焦荧光显微镜进行单分子观察。具体地，针对前哨淋巴结中的多个区域中的每个区域测量荧光强度，并且在测量的多个区域中，以荧光强度的降序将一个或多个区域作为传入淋巴管流入区域检测。

9. 发明授权

US09976959B2 Biological substance detection method using florescent dye-containing particle 有权
公开(公告)号：US09976959B2
公开(公告)日：2018-05-22
申请号：US13819453
申请日：2011-08-30
申请人： Kensaku Takanashi , Hideki Goda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
发明人： Kensaku Takanashi , Hideki Goda , Hisatake Okada , Yasushi Nakano , Kohsuke Gonda , Motohiro Takeda , Noriaki Ohuchi
IPC分类号： G01N21/64 , G01N33/483 , G01N33/58
CPC分类号： G01N21/6486 , G01N21/6428 , G01N21/6458 , G01N33/4833 , G01N33/582 , G01N2021/6439 , G01N2021/6441
摘要： A biological substance detection method for detecting a biological substance specifically in a pathological specimen, comprising a step of immunologically staining the pathological specimen using a fluorescent label, a step of staining the pathological specimen with a staining reagent for morphology observation purposes (eosin) to observe the morphology of the pathological specimen, a step of irradiating the stained pathological specimen with excited light to cause the emission of a fluorescent and detecting the biological substance in the pathological specimen. In the step of immunologically staining the pathological specimen, a special fluorescent particle for which the excitation wavelength appears in a region that is different from the excitation wavelength region of eosin is used as the fluorescent label.

10. 发明申请

US20130203082A1 METHOD FOR DETERMINING CANCER ONSET OR CANCER ONSET RISK 审中-公开
标题翻译：用于确定癌症或癌症风险的方法
公开(公告)号：US20130203082A1
公开(公告)日：2013-08-08
申请号：US13819008
申请日：2011-08-26
申请人： Kohsuke Gonda , Minoru Miyashita , Motohiro Takeda , Noriaki Ohuchi
发明人： Kohsuke Gonda , Minoru Miyashita , Motohiro Takeda , Noriaki Ohuchi
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , B82Y15/00 , G01N21/6428 , G01N21/6458 , G01N33/57415 , G01N33/588 , G01N2800/50
摘要： A highly accurate and quantitative method for determining cancer onset or cancer onset risk by a quantitative tissue staining method in biological tissues using an antibody capable of recognizing a cancer growth regulatory factor or cancer metastasis regulatory factor such as PAR1 antibody, which inhibits the cancer cell mobility and infiltration is provided. Cancer onset or cancer onset risk is determined using the tissue staining method comprising the steps of: labeling an antibody which recognizes a cancer growth regulatory factor or cancer metastasis regulatory factor with a fluorescent material, and contacting the fluorescent-labeled antibody with a tissue sample; irradiating a tissue site in contact with the antibody with excitation light to acquire a fluorescence image; acquiring an autofluorescence image in a vicinity region of a short wavelength side or long wavelength side of an acquisition region of fluorescence wavelength emitted by the fluorescent material, in the same field of vision and in the same focal point as those of the fluorescence image; acquiring a corrected fluorescence image by image processing to eliminate a fluorescent brightness of the autofluorescence image from the fluorescent brightness of the fluorescence image; counting the number of cells at the tissue site in contact with the antibody; measuring a mean fluorescent brightness of a single fluorescent particle; and calculating the number of fluorescent particles per cell.
摘要翻译：使用能够识别癌细胞生长调节因子或癌转移调节因子（例如抑制癌细胞迁移率的PAR1抗体）的抗体，通过定量组织染色法在生物组织中测定癌症发病或癌症发病风险的高度准确和定量的方法并提供渗透。使用组织染色方法确定癌症发病或癌症发病风险，其包括以下步骤：用荧光材料标记识别癌症生长调节因子或癌症转移调节因子的抗体，并将荧光标记的抗体与组织样品接触; 用激发光照射与抗体接触的组织部位以获得荧光图像; 在荧光材料发射的荧光波长的获取区域的短波长侧或长波长侧的附近区域中，在与荧光图像相同的视场和相同的焦点中获取自发荧光图像; 通过图像处理获得校正的荧光图像，以从荧光图像的荧光亮度消除自发荧光图像的荧光亮度; 计数与抗体接触的组织部位的细胞数; 测量单个荧光颗粒的平均荧光亮度; 并计算每个细胞的荧光颗粒数。

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