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1. 发明专利

JP2021519448A 散乱を利用した超局在顕微鏡法およびその関連装置审中-公开
公开(公告)号：JP2021519448A
公开(公告)日：2021-08-10
申请号：JP2020552880
申请日：2019-03-26
申请人：クレストオプティクスエス.ピー.エー. , CRESTOPTICS S.P.A. , フォンダツィオーネ・イスティトゥート・イタリアーノ・ディ・テクノロジャ , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人：レオネッティマルコ , アントナッチジュゼッペ , チェッカレッリライノ
IPC分类号： G01N21/64 , G02B21/00
摘要：本発明は、蛍光顕微鏡法であって、A. 第1の初期構成C 1 をもつ波面を有するコヒーレント励起光線(5)を備え、1つ以上の蛍光発光体を含む散乱サンプルを照らすことで、当該散乱サンプルに含まれる1つ以上の蛍光発光体を励起するステップと、B. 画素撮像ユニット(45)を介して、散乱サンプルの背景画像F 1 =F B に対応する第1の初期画像F 1 を取得し、スペックル粒子画像を得るステップと、C. 少なくとも1つの蛍光発光体の蛍光シグナルをもつ第1の初期画像F 1 の領域に属するN=1…M個の座標[x N ,y N ]を有し、それぞれが第1の初期強度I PN 1 をもつM個の目標ピクセルP N を選択するステップと、D. M個の目標ピクセルP N のそれぞれについて波面の第1の初期構成を最適化することで、第1の初期画像F 1 のスペックル粒子サイズを減少させ、目標ピクセルP N の最大強度に対応する蛍光シグナルの局所最大I PN max を有する散乱サンプルの最終画像F fin (I PN max )を取得するステップと、E. ステップBで取得されたバックグラウンドの第1の初期画像F B を、ステップDで取得された最終画像F fin (I PN max )から減算することで、M個の目標ピクセルP N のそれぞれについて、強度I PN SPECKLE で画素[x N ,y N ]に位置する1つのスペックル粒子から来る蛍光シグナルのみを有する画像F speckle(PN) をそれぞれ取得するステップと、F. 自由中心座標をもつガウス関数で、ステップEで得られた画像F speckle(PN) をフィッティングすることで、M個の目標ピクセルP N のそれぞれについて、xy平面での座標(X N ,Y N )と強度I N を得るステップと、G. M個の目標ピクセルP N のそれぞれについて、ステップFで得られた強度I N と、散乱サンプルのスペックル粒子の平均サイズSに等しいクビレをもち、座標(X N ,Y N )を中心とするガウス分布を含む画像F N を生成するステップと、H. M個の目標ピクセルP N のそれぞれについて、ステップGで得られたM個の画像F N を足し合わせてサンプルの最終画像を生成するステップと、を含む。【選択図】図3

2. 发明申请

US20210063721A1 SCATTERING-ASSISTED SUPER-LOCALIZATION MICROSCOPY METHOD AND RELATIVE APPARATUS 有权
公开(公告)号：US20210063721A1
公开(公告)日：2021-03-04
申请号：US17040301
申请日：2019-03-26
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： Marco LEONETTI , Giuseppe ANTONACCI , Raino CECCARELLI
IPC分类号： G02B21/36 , G01N21/64 , G02B21/16
摘要： Fluorescence microscopy method comprising: illuminating a scattering sample with a coherent excitation light beam having a wavefront with an initial configuration for exciting fluorescent emitters in the scattering sample; acquiring an initial image; selecting target pixels in an area of the initial image; optimising the initial configuration of the wavefront for each target pixel for decreasing the speckle grain size and obtaining a final image; subtracting the initial image from the final image for each target pixel, obtaining an image; fitting the image with a Gaussian function with free center coordinates for each target pixel; generating an image containing a Gaussian distribution centered at the coordinates and intensity, and with a waist equal to an average size S of speckle grain of the scattering sample, for each target pixel; and generating a final image of the sample by summing the images of the target pixels.

3. 发明申请

US20200333305A1 APPARATUS AND METHOD FOR AUTOMATICALLY DETECTING ODORANT SUBSTANCES IN SOLUTION BY USING NEMATODES CAENORABDITIS ELEGANS 审中-公开
公开(公告)号：US20200333305A1
公开(公告)日：2020-10-22
申请号：US16756669
申请日：2018-10-19
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： Viola FOLLI , Andrea SANTINELLI , Marco BROGLIA
IPC分类号： G01N33/00 , B01L3/00 , G01N33/50 , G01N21/64
摘要： Apparatus (1000) and method for automatically detecting odorant substances based on use of nematodes that includes a mechanical selection unit (100) configured to select nematodes in adult stage from an initial nematode population obtaining an intermediate nematode population, a nematode optical selection unit (200) configured to select from the intermediate population a final population of nematodes in adult stage and to select nematodes in young adult stage from nematodes in egg producing adult stage to be sent to a measurement unit (300) configured to detect the response of nematodes of the final population to a stimulus of an odorant substance, the mechanical selection unit (100) being connected to the optical selection unit (200) by a connection channel with an at least three way branch and the optical selection unit (200) being connected to the measurement unit (300) by a loading microchannel.

4. 发明授权

US10905324B2 Spatial super-resolution apparatus for fluorescence analysis of eye fundus 有权
公开(公告)号：US10905324B2
公开(公告)日：2021-02-02
申请号：US16461359
申请日：2017-11-21
申请人： CRESTOPTICS S.p.A.
发明人： Vincenzo Ricco , Raino Ceccarelli , Andrea Latini
IPC分类号： A61B3/125 , A61B3/00 , A61B3/13 , A61B3/14 , G02B21/00 , G02B21/36
摘要： Fluorescence microscopy apparatus for the analysis of a fundus of a sample eye, based on the use of an optical equipment configured to generate a Bessel beam inside a sample eye and an objective configured to increase the numerical aperture of an “objective-cornea-lens” assembly by reducing the overall focal length of the fluorescence microscopy apparatus, also allowing a significant increase in spatial resolution compared to conventional microscopy systems.

5. 发明申请

US20190320894A1 SPATIAL SUPER-RESOLUTION APPARATUS FOR FLUORESCENCE ANALYSIS OF EYE FUNDUS 审中-公开
公开(公告)号：US20190320894A1
公开(公告)日：2019-10-24
申请号：US16461359
申请日：2017-11-21
申请人： CRESTOPTICS S.p.A.
发明人： Vincenzo Ricco , Raino Ceccarelli , Andrea Latini
IPC分类号： A61B3/125 , A61B3/13 , A61B3/14 , A61B3/00 , G02B21/00 , G02B21/36
摘要： Fluorescence microscopy apparatus for the analysis of a fundus of a sample eye, based on the use of an optical equipment configured to generate a Bessel beam inside a sample eye and an objective configured to increase the numerical aperture of an “objective-cornea-lens” assembly by reducing the overall focal length of the fluorescence microscopy apparatus, also allowing a significant increase in spatial resolution compared to conventional microscopy systems.

6. 发明申请

US20180292634A1 CONFOCAL MICROSCOPY APPARATUS AND RELATED PROCESS FOR ACQUIRING AND PROCESSING IMAGES 审中-公开
公开(公告)号：US20180292634A1
公开(公告)日：2018-10-11
申请号：US15764332
申请日：2016-10-12
申请人： CRESTOPTICS S.p.A.
发明人： Vincenzo RICCO , Andrea SANTINELLI
IPC分类号： G02B21/00 , G02B21/36 , G06T3/40
摘要： Confocal microscopy apparatus, and related process, comprising: a structured light generating component configured to be illuminated with a basic light beam and to generate a structured light beam focused on a first plane; a spinning disk configured to receive said structured light beam and to transmit a resulting excitation beam to an optics of a microscope focused on a plane of a sample, wherein the spinning disk lies on a second plane and comprises a disk-shaped substrate composed of an optically transparent material, the substrate of the spinning disk comprising a planar first surface and an opposing planar second surface and a patterned mask disposed on one of the first surface and the second surface and comprising at least one sector provided with one or more continuous spiral slit apertures, wherein the patterned mask or an outer surface thereof is composed of a highly black material opaque to light; a housing configured to house on a third plane an acquisition sensor configured to detect a fluorescent beam emitted from said plane of the sample; a set of relay lenses configured to optically conjugate the first plane, the second plane and said plane of the sample to the third plane; optical means configured to transmit said structured light beam from the structured light generating component to said plane of the sample and said emitted fluorescent beam from said plane of the sample to said housing configured to house the acquisition sensor; and moving means configured to move the structured light generating component, so as to shift the structured light beam in the first plane, and the spinning disk in the second plane.

7. 发明申请

WO2021240319A1 MICROFLUIDIC METHOD FOR THE DETECTION OF NUCLEIC ACIDS 审中-公开
公开(公告)号：WO2021240319A1
公开(公告)日：2021-12-02
申请号：PCT/IB2021/054419
申请日：2021-05-21
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： RICCO, Vincenzo , CECCARELLI, Raino , BOFFI, Alberto , RUOCCO, Giancarlo
IPC分类号： C12Q1/6816
摘要： The present invention describes a novel method for the fast detection of an analyte, typically a viral nucleic acid. The method relies on the denaturation of the viral particle and immediate mixing with pairing primers (complementary oligonucleotides) linked to a fluorophore. The RNA containing solution and the fluorescent primers containing solution are then mixed in a v-shaped conduit under centrifugal force provided by a spinning disk. Once mixed, the reactants are heated and forced through a molecular sieve that separates larger viral nucleic acid complexed with fluorescent primers from unreacted oligonucleotides. Labelled viral molecules are then probed by a focalized laser light at the absorption wavelength of the fluorophore while physically trapped into the molecular sieve.

8. 发明申请

WO2019186393A1 SCATTERING-ASSISTED SUPER-LOCALIZATION MICROSCOPY METHOD AND RELATIVE APPARATUS 审中-公开
公开(公告)号：WO2019186393A1
公开(公告)日：2019-10-03
申请号：PCT/IB2019/052442
申请日：2019-03-26
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： LEONETTI, Marco , ANTONACCI, Giuseppe , CECCARELLI, Raino
IPC分类号： G01N21/64 , G02B21/00
摘要： Fluorescence microscopy method comprising the following steps: A. illuminating a scattering sample including one or more fluorescent emitters with a coherent excitation light beam (5) having a wavefront with a first initial configuration C 1 for exciting said one or more fluorescent emitters comprised in the scattering sample; B. acquiring a first initial image F 1 corresponding to a background image F 1 =F 8 of the scattering sample, through a pixel imaging unit (45), obtaining a speckle grain image; C. selecting M target pixels P N , with N = 1,…,M, of coordinates [X N , Z N ] belonging to an area of the first initial image F 1 having fluorescence signals of at least one fluorescent emitters, each one of the M target pixels P N having a first initial intensity I PN 1 ; D. optimising the first initial configuration of the wavefront for each one of the M target pixels P N , for decreasing the speckle grain size of the first initial image F 1 and obtaining a final image F fin (I PN max ) of the scattering sample with a local maximum I PN max of the fluorescence signal corresponding to a maximum intensity of the target pixels P N ; E. subtracting the first initial image F B of background, obtained at step B, from the final image F fin (I PN max ) , obtained at step D, for each one of the M target pixels P N , obtaining an image F speckle ( PN ) having only fluorescent signals coming from a speckle grain located at pixel [X N , Z N ] with intensity I PN speckle , for each one of the M target pixels P N ; F. fitting the image F speckle ( PN ) , obtained at step E, with a Gaussian function with free center coordinates, for each one of the M target pixels P N , thereby obtaining coordinates ( X N , Z N ) and intensity I N in a xy plane; G. generating an image F N containing a Gaussian distribution centered at coordinates ( X N , Z N ) and with intensity I N , obtained at step F, and with a waist equal to an average size S of speckle grain of the scattering sample, for each one of the M target pixels P N ; H. generating a final image of the sample by summing the M images F N obtained at step G, for each one of the M target pixels P N ;

9. 发明申请

WO2021191767A1 MICROFLUIDIC DEVICE TO DETECT THE NEURONAL ACTIVITY OF A MICRO-ORGANISM 审中-公开
公开(公告)号：WO2021191767A1
公开(公告)日：2021-09-30
申请号：PCT/IB2021/052355
申请日：2021-03-22
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： FOLLI, Viola , CAPRINI, Davide , CECCARELLI, Raino
IPC分类号： C12Q1/02 , B01L3/00 , C12M1/34 , C12M3/00 , B01L2200/0668 , B01L2300/0877 , B01L3/502746 , B01L3/502761 , B01L3/502776 , C12M23/16 , C12Q1/025
摘要： The present invention relates to a microfluidic device (100) for a system of detecting the neuronal activity of a population of sample micro-organisms, which device (10) comprises: - a conduit (10) comprising an intermediate portion (13) arranged between an inlet portion (11) and an outlet portion (12) of an operating fluid in/from the conduit (10), said conduit (10) being shaped so as to receive the operating fluid crossing through a longitudinal direction (A), - a stagnation chamber (20) configured to receive internally the population of sample micro-organisms, wherein said stagnation chamber (20) is in fluid communication with said conduit (10) through one or more openings (30) obtained in the intermediate portion (13) and configured to keep unchanged, in use, the crossing direction of the operating fluid, and wherein the ratio between the cross section passage (o) of the operating fluid in the conduit (10) at said intermediate portion (13) and the cross section passage of the operating fluid in the conduit (10) at said inlet portion (11) or at said outlet portion (12) is greater than 1.

10. 发明申请

WO2021240209A1 DEVICE AND METHOD FOR DETECTING A TARGET MOLECULE IN A BIOLOGICAL FLUID 审中-公开
公开(公告)号：WO2021240209A1
公开(公告)日：2021-12-02
申请号：PCT/IB2020/054966
申请日：2020-05-26
申请人： CRESTOPTICS S.P.A. , FONDAZIONE ISTITUTO ITALIANO DI TECNOLOGIA
发明人： RICCO, Vincenzo , CECCARELLI, Raino , BOFFI, Alberto , RUOCCO, Giancarlo
IPC分类号： B01L3/00 , F16K99/00
摘要： A testing device (1) for detecting a target molecule in a biological fluid sample, which device comprises: ▪ a disk-shaped main body (100), configured to be rotatable about a central spinning axis (110); ▪ a microfluidic path (10) obtained on said main body (100), which microfluidic path defines a testing radial sector, the latter comprising: - a sample reservoir (A) receiving the biological fluid sample; - a reactant reservoir (B) receiving a reactant apt to bind with the target molecule; - a detection chamber (160), arranged at a greater radial distance from the spinning axis (110) with respect to said sample reservoir (A) and reactant reservoir (B); - conduits (131, 132) connecting the sample reservoir (A) and the reactant reservoir (B) to the detection chamber (160) for adducting thereto their contents, wherein rotation of the main body (100) determines the adduction of the biologic fluid sample and of the reactant from their respective reservoirs (A, B) towards said detection chamber (160) by centrifugal force.

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