公开(公告)号：US20140178927A1

公开(公告)日：2014-06-26

申请号：US14118150

申请日：2012-05-18

申请人： Atsushi Miyawaki ,  Hiroshi Hama ,  Asako Sakaue

发明人： Atsushi Miyawaki ,  Hiroshi Hama ,  Asako Sakaue

IPC分类号： G01N1/30

CPC分类号： G01N1/30 ,  G01N33/5002 ,  G01N33/5005 ,  G01N33/5008 ,  G01N2001/302

摘要： A clearing reagent according to the present invention for making a biological material transparent is a solution containing: at a concentration of 1M or more and not more than 8.5M, at least one compound selected from the group consisting of urea and urea derivatives; and glycerol at a concentration of 25 (w/v) % or more and not more than 35 (w/v) %.

摘要翻译： 根据本发明的用于使生物材料透明的清洗试剂是含有浓度为1M以上至不超过8.5M的至少一种选自脲和脲衍生物的化合物的溶液; 和浓度为25（w / v）％以上且不高于35（w / v）％的甘油。

公开(公告)号：US20130045503A1

公开(公告)日：2013-02-21

申请号：US13583548

申请日：2011-03-11

申请人： Atsushi Miyawaki ,  Hiroshi Hama ,  Hiroshi Kurokawa ,  Hiroyuki Kawano ,  Asako Sakaue

发明人： Atsushi Miyawaki ,  Hiroshi Hama ,  Hiroshi Kurokawa ,  Hiroyuki Kawano ,  Asako Sakaue

IPC分类号： G01N1/30

CPC分类号： G01N1/30

摘要： A clearing reagent according to this invention for making a biological material transparent is a solution containing, as an active component, at least one compound selected from the group consisting of urea and urea derivatives, in order to provide a novel clearing reagent for making a biological material transparent.

摘要翻译： 根据本发明的用于使生物材料透明化的清洗试剂是含有选自脲和脲衍生物的至少一种化合物作为活性组分的溶液，以提供用于制备生物材料的新型清除试剂 材料透明。

公开(公告)号：US20110206615A1

公开(公告)日：2011-08-25

申请号：US12920283

申请日：2009-02-27

申请人： Atsushi Miyawaki ,  Kazuki Sasaki

发明人： Atsushi Miyawaki ,  Kazuki Sasaki

IPC分类号： A61K49/00 ,  C12N15/63 ,  G01N33/53 ,  C12Q1/68 ,  C07K19/00 ,  C12N9/02 ,  C07H21/04 ,  C07H21/02

CPC分类号： G01N33/5008 ,  A61K49/0017 ,  A61K49/0056 ,  A61K49/14 ,  A61K51/08 ,  A61K51/088 ,  C07K14/00 ,  C12Q1/28 ,  G01N21/6428 ,  G01N21/77 ,  G01N33/5014 ,  G01N33/84 ,  G01N2500/10

摘要： The present invention relates to fluorescent or luminescent probe reagents for measuring oxidative stress in a cell or an organism. Examples of the probe reagents include: a fluorescent or luminescent protein and a marker protein; a fluorescent or luminescent protein, a marker protein, and a regulatory factor; or a fluorescent or luminescent protein, a marker protein, a cleavage sequence, and a regulatory factor. In the probe reagents, the marker protein makes it possible to detect the oxidative stress caused by reactive oxygen species and comprises a regulatory factor-binding site and a ubiquitin-binding site; and the regulatory factor is a protein making it possible to regulate degradation of the marker protein in response to the reactive oxygen species. The present invention also relates to a method of measuring oxidative stress in a cell or an organism, or a method of screening a substance which suppresses or promotes the oxidative stress in a cell or an organism by using the probe reagent.

摘要翻译： 本发明涉及用于测量细胞或生物体中的氧化应激的荧光或发光探针试剂。 探针试剂的实例包括：荧光或发光蛋白和标记蛋白; 荧光或发光蛋白，标记蛋白和调节因子; 或荧光或发光蛋白，标记蛋白，切割序列和调节因子。 在探针试剂中，标记蛋白使得可以检测由活性氧引起的氧化应激，并且包含调节因子结合位点和泛素结合位点; 并且调节因子是使得可以响应于活性氧来调节标记蛋白的降解的蛋白质。 本发明还涉及一种测定细胞或生物体中的氧化应激的方法，或者通过使用探针试剂筛选抑制或促进细胞或生物体中的氧化应激的物质的方法。

公开(公告)号：US07595375B2

公开(公告)日：2009-09-29

申请号：US10162593

申请日：2002-06-06

申请人： Atsushi Miyawaki ,  Takeharu Nagai

发明人： Atsushi Miyawaki ,  Takeharu Nagai

IPC分类号： C07K14/00 ,  C07H21/04 ,  C12N1/20 ,  C12N15/00

CPC分类号： C07K14/43595 ,  C07K2319/00

摘要： The present invention provides a fluorescent protein having an amino acid sequence of the green fluorescent protein, the yellow fluorescent protein or mutants thereof wherein the 46th phenylalanine residue is substituted with a leucine residue. According to the present invention, there are provided novel GFP or YEP mutants having an excellent maturation efficacy and having a decreased sensitivity to both of H+ and Cl−.

摘要翻译： 本发明提供了具有绿色荧光蛋白的氨基酸序列，黄色荧光蛋白或其突变体的荧光蛋白，其中第46位苯丙氨酸残基被亮氨酸残基取代。 根据本发明，提供了具有优异成熟功效并且对H +和Cl-两者具有降低的敏感性的新型GFP或YEP突变体。

公开(公告)号：US20090176211A1

公开(公告)日：2009-07-09

申请号：US11719166

申请日：2005-11-14

申请人： Takeharu Nagai ,  Atsushi Miyawaki

发明人： Takeharu Nagai ,  Atsushi Miyawaki

IPC分类号： C12Q1/68 ,  C07K14/00 ,  C12N15/11 ,  C12N15/00 ,  C12N5/06

CPC分类号： G01N33/5035 ,  C07K14/43595 ,  G01N33/542

摘要： It is an object of the present invention to provide a fluorescent protein, which allows an acceptor for fluorescence resonance energy transfer (FRET) to appear in a stimulating light-dependent manner, thereby enabling the marking of any given cell organelle, cells, or tissues, with multiple colors. The present invention provides a fluorescent protein which consists of a fused protein of a donor fluorescent protein and an acceptor fluorescent protein, wherein before irradiation with stimulating light, the donor protein is able to emit fluorescence as a result of irradiation of the donor protein with excitation light; and after irradiation with stimulating light, intramolecular FRET occurs between the donor fluorescent protein and the acceptor fluorescent protein as a result of irradiation of the donor protein with excitation light, and the acceptor protein is able to emit fluorescence, and wherein the fluorescence of the donor protein and the fluorescence of the acceptor protein have wavelengths that are different from each other.

摘要翻译： 本发明的目的是提供一种荧光蛋白，其允许荧光共振能量转移受体（FRET）以刺激光依赖的方式出现，从而能够标记任何给定的细胞器，细胞或组织 ，具有多种颜色。 本发明提供了由供体荧光蛋白的融合蛋白和受体荧光蛋白组成的荧光蛋白，其中在用刺激光照射之前，供体蛋白能够通过激发照射供体蛋白而发出荧光 光; 并且在用刺激光照射后，作为供体蛋白质与激发光照射的结果，在供体荧光蛋白和受体荧光蛋白之间发生分子内FRET，并且受体蛋白能够发射荧光，并且其中供体的荧光 蛋白质，受体蛋白质的荧光具有彼此不同的波长。

公开(公告)号：US20090017516A1

公开(公告)日：2009-01-15

申请号：US11722283

申请日：2005-12-21

申请人： Takeharu Nagai ,  Atsushi Miyawaki

发明人： Takeharu Nagai ,  Atsushi Miyawaki

IPC分类号： C12N13/00 ,  C07K14/00 ,  C07F19/00 ,  C07K1/00

CPC分类号： C07K14/43595 ,  A61K41/0057 ,  A61K41/0071

摘要： An object of the present invention is to provide a method of generating reactive oxygen species in a light irradiation-dependent manner, so as to inactivate any target physiological function. The present invention provides a target physiological function inactivator which consists of a photosensitizer-labeled fluorescent protein, wherein fluorescence resonance energy transfer (FRET) from the fluorescent protein to the photosensitizer occurs as a result of light irradiation, so that the photosensitizer can be excited to generate reactive oxygen species.

摘要翻译： 本发明的目的是提供一种依照光依赖的方式产生活性氧的方法，从而使任何目标生理功能失活。 本发明提供了由光敏剂标记的荧光蛋白组成的目标生理功能失活剂，其中由于光照射而发生从荧光蛋白到光敏剂的荧光共振能量转移（FRET），使得光敏剂可被激发 产生活性氧。

公开(公告)号：US07277566B2

公开(公告)日：2007-10-02

申请号：US10662355

申请日：2003-09-16

申请人： Atsushi Miyawaki ,  Takashi Fukano

发明人： Atsushi Miyawaki ,  Takashi Fukano

IPC分类号： G06K9/00

CPC分类号： G02B21/082 ,  G01B11/24 ,  G01B11/25

摘要： A microscope system comprises: a control means (18) capable of generating electric control signals (18a); a spatial modulator means (7) having an illuminated surface (7a) to be illuminated by light emitted by a light source (1), and capable of receiving the electric control signal and of spatially modulating reflection characteristic or transmission characteristic of the illuminated surface by a spatial frequency specified by the electric control signal; an illuminating optical means (8, 11) for illuminating a specimen (12) with light spatially modulated by the spatial modulator means; an image detecting means (15) for detecting a signal image formed by signal light emitted by the specimen illuminated by the illuminating optical means; and an arithmetic means (16) for processing signal images formed by using the spatial frequency of at least three different phases set by the control means and detected by the image detecting means to obtain an optical sectioned image.

摘要翻译： 显微镜系统包括：能够产生电控信号的控制装置（18）; 空间调制器装置（7）具有被光源（1）发射的光照射的照明表面（7a），并且能够接收电控制信号和空间调制被照射表面的反射特性或透射特性 通过由电控信号指定的空间频率; 用于通过由空间调制器装置空间调制的光照射样本（12）的照明光学装置（8,11）; 图像检测装置，用于检测由照明光学装置照射的样本发出的信号光形成的信号图像; 以及用于处理通过使用由控制装置设置并由图像检测装置检测的至少三个不同相位的空间频率形成的信号图像以获得光学分割图像的算术装置（16）。

公开(公告)号：US07236298B2

公开(公告)日：2007-06-26

申请号：US11520150

申请日：2006-09-13

申请人： Atsushi Miyawaki ,  Takashi Fukano ,  Yasushi Aono

发明人： Atsushi Miyawaki ,  Takashi Fukano ,  Yasushi Aono

IPC分类号： G02B21/06

CPC分类号： G02B21/06 ,  G02B21/16 ,  G02B21/365

摘要： An illumination apparatus for a microscope and an image processing apparatus using the illumination apparatus include a light source, a semi-transmissive mirror splitting a light beam from the light source into two beams of the first and second irradiation light, two excitation filters selecting the wavelengths of the first and second irradiation light, a semi-transmissive mirror synthesizing individual beams of the first and second irradiation light whose wavelengths are selected, into a single beam, a dichroic mirror directing a light beam synthesized by the semi-transmissive mirror toward a specimen and transmitting light from the specimen, an objective lens, cameras imaging fluorescent light from the specimen after being separated into fluorescent light excited by the first and second wavelengths, and an image processing section processing fluorescent images formed by imaging elements.

摘要翻译： 用于显微镜的照明装置和使用该照明装置的图像处理装置包括光源，将来自光源的光束分成两束第一和第二照射光的半透射镜，两个激发滤光器选择波长 第一和第二照射光的第一和第二照射光的半透射镜将将波长选择的第一和第二照射光的各个光束合成为单个光束，将由半透射镜合成的光束引向样品的二向色镜 以及透射来自所述样本的物体，物镜，将被分离成由所述第一和第二波长激发的荧光的荧光从所述样本成像的摄像机，以及处理由成像元件形成的荧光图像的图像处理部。

公开(公告)号：US20070087436A1

公开(公告)日：2007-04-19

申请号：US10552923

申请日：2004-04-09

申请人： Atsushi Miyawaki ,  Hiroyuki Imabayashi ,  Sachiko Karaki

发明人： Atsushi Miyawaki ,  Hiroyuki Imabayashi ,  Sachiko Karaki

IPC分类号： C12N15/00 ,  C12M1/00

CPC分类号： B82Y30/00 ,  C12M35/00

摘要： An object of the present invention is to provide a method for introducing a physiologically active substance such as a gene into a cell, which introduces a physiologically active substance such as any given gene into any given cell in a view under a microscope, while significantly reducing invasiveness to the cell, and a device used for the above method. The present invention provides a method for introducing a physiologically active substance into a cell, which comprises: allowing a physiologically active substance to attach around a needle having a diameter of 500 nm or less, provided that it is able to be inserted into a cell; and inserting the above-described needle into the cell; and a microinjection device for carrying out the aforementioned method.

摘要翻译： 本发明的目的是提供一种将生物活性物质如基因导入细胞的方法，该细胞在显微镜下以任何给定的基因引入任何给定基因的生理活性物质，同时显着降低 对细胞的侵袭性，以及用于上述方法的装置。 本发明提供了一种将生理活性物质引入细胞的方法，其包括：使生理活性物质附着在直径为500nm以下的针上，只要其能够插入细胞中即可。 并将上述针插入细胞中; 以及用于实施上述方法的显微注射装置。

公开(公告)号：US20070024964A1

公开(公告)日：2007-02-01

申请号：US11529017

申请日：2006-09-28

申请人： Kiyochika Ishibashi ,  Atsushi Miyawaki

发明人： Kiyochika Ishibashi ,  Atsushi Miyawaki

IPC分类号： G02B21/00

CPC分类号： G01N21/6408 ,  G01N21/6428 ,  G01N2021/6419 ,  G01N2021/6441

摘要： There is provided a fluorescence spectroscopic apparatus includes an exciting optical unit configured to irradiate the same sample area with a plurality of excitation lights of different wavelength bands, an optical unit configured to repeatedly guide fluorescences emitted by the sample in response to the respective excitation lights, to a detection unit, and a calculation unit configured to perform analysis on the basis of a comparison of output signals corresponding to the fluorescences from the detection unit, wherein the exciting optical unit includes an excitation light varying unit configured to intermittently vary the intensity of at least one excitation light.

摘要翻译： 提供了一种荧光分光装置，包括：激发光学单元，被配置为用不同波长带的多个激发光照射相同的样本区域;光学单元，被配置为响应于各个激发光反复引导样品发射的荧光， 检测单元和计算单元，其被配置为基于与来自检测单元的荧光相对应的输出信号的比较来执行分析，其中激发光学单元包括激励光改变单元，其被配置为间歇地改变在 至少一个激发光。