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公开(公告)号：US10247672B2
公开(公告)日：2019-04-02
申请号：US14869968
申请日：2015-09-29
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Dong Li
Abstract: A method includes: (a) providing spatially-patterned activation radiation to a sample that includes phototransformable labels, where an optical parameter of the spatially-patterned activation radiation varies periodically in space; (b) providing spatially-patterned excitation radiation to the sample, where an optical parameter of the spatially-patterned excitation radiation varies periodically in space, where (a) and (b) create a non-linear fluorescence emission pattern within the sample, the pattern including H modulation harmonics, with H>1. The method includes (c) detecting radiation emitted from the activated and excited labels, (d) storing detected radiation data, and (e) spatially shifting one or both of the spatially-patterned excitation radiation and the spatially-patterned activation radiation with respect to the sample to spatially shift the non-linear fluorescence emission pattern within the sample, and (f) repeating (a)-(e) at least N times, with N>2. Then, a sub-diffraction-limited final image of the sample is generated based on the stored data for the N positions of the non-linear fluorescence emission pattern within the sample.
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公开(公告)号：US20180210184A1
公开(公告)日：2018-07-26
申请号：US15843633
申请日：2017-12-15
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig
Abstract: A microscope has a light source for generating a light beam having a wavelength, λ, and beam-forming optics configured for receiving the light beam and generating a Bessel-like beam that is directed into a sample. The beam-forming optics include an excitation objective having an axis oriented in a first direction. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beam and for imaging the received light on a detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction. A detector is configured for detecting signal light received by the imaging optics, and an aperture mask is positioned.
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公开(公告)号：US20150362713A1
公开(公告)日：2015-12-17
申请号：US14660906
申请日：2015-03-17
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Kai Wang
CPC classification number: G02B21/0064 , G01N21/6458 , G01N2021/6419 , G01N2201/0675 , G02B21/0056 , G02B21/006 , G02B21/0072 , G02B21/0076 , G02B21/008 , G02B21/361 , G02B26/06
Abstract: Excitation light is focused to a focus within a sample and the focus is scanned within a volume in the sample with scanning optical elements. Signal light emitted from the focus is de-scanned, with the one or more scanning optical elements, onto a wavefront sensor as the focus is scanned within the volume. Based on the descanned signal light, an average aberration created by the volume of the sample of a wavefront of the excitation light is determined. A wavefront of the excitation light is corrected by an amount according to the determined average aberration while the focus is scanned within the volume, the signal light is imaged onto a photosensitive detector as the focus is scanned within the volume, and a wavefront of the imaged signal light is corrected by an amount according to the determined average aberration while the focus is scanned. These steps can be repeated for a plurality of different volumes in the sample, and an image of the sample can be generated based on the detected signal light from scanned foci within the different volumes.
Abstract translation: 激发光聚焦到样品中的焦点，并用扫描光学元件在样品中的体积内扫描焦点。 当聚焦在体积内被扫描时，从焦点发出的信号光与一个或多个扫描光学元件一起被扫描到波前传感器上。 基于下拉信号光，确定由激发光的波前的样本的体积产生的平均像差。 当在体积内扫描焦点时，根据确定的平均像差来校正激发光的波前，当体积内的焦点被扫描时，信号光被成像到光敏检测器上，并且成像的波前 当扫描焦点时，根据确定的平均像差校正信号光的量。 可以对样本中的多个不同体积重复这些步骤，并且可以基于来自不同体积内的扫描焦点的检测到的信号光来生成样本的图像。
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公开(公告)号：US10051240B2
公开(公告)日：2018-08-14
申请号：US13844405
申请日：2013-03-15
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Kai Wang
Abstract: An apparatus includes a light source configured for generating a coherent light beam having a wavelength, λ, a light detector, and beam-forming optics configured for receiving the generated light beam and for generating a plurality of substantially parallel Bessel-like beams directed into a sample in a first direction. Each of the Bessel-like beams has a fixed phase relative to the other Bessel-like beams. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beams and for imaging the received light onto the detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction, where the detector is configured for detecting light received by the imaging optics. A processor configured to generate an image of the sample based on the detected light.
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公开(公告)号：US09500846B2
公开(公告)日：2016-11-22
申请号：US14660906
申请日：2015-03-17
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Kai Wang
CPC classification number: G02B21/0064 , G01N21/6458 , G01N2021/6419 , G01N2201/0675 , G02B21/0056 , G02B21/006 , G02B21/0072 , G02B21/0076 , G02B21/008 , G02B21/361 , G02B26/06
Abstract: Excitation light is focused to a focus within a sample and the focus is scanned within a volume in the sample with scanning optical elements. Signal light emitted from the focus is de-scanned, with the one or more scanning optical elements, onto a wavefront sensor as the focus is scanned within the volume. Based on the descanned signal light, an average aberration created by the volume of the sample of a wavefront of the excitation light is determined. A wavefront of the excitation light is corrected by an amount according to the determined average aberration while the focus is scanned within the volume, the signal light is imaged onto a photosensitive detector as the focus is scanned within the volume, and a wavefront of the imaged signal light is corrected by an amount according to the determined average aberration while the focus is scanned. These steps can be repeated for a plurality of different volumes in the sample, and an image of the sample can be generated based on the detected signal light from scanned foci within the different volumes.
Abstract translation: 激发光聚焦到样品中的焦点，并用扫描光学元件在样品中的体积内扫描焦点。 当聚焦在体积内被扫描时，从焦点发出的信号光与一个或多个扫描光学元件一起被扫描到波前传感器上。 基于下拉信号光，确定由激发光的波前的样本的体积产生的平均像差。 当在体积内扫描焦点时，根据确定的平均像差校正激发光的波前，当体积内的焦点被扫描时，将信号光成像到光敏检测器上，并且成像的波前 当扫描焦点时，根据确定的平均像差校正信号光的量。 可以对样本中的多个不同体积重复这些步骤，并且可以基于来自不同体积内的扫描焦点的检测到的信号光来生成样本的图像。
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Patent Agency Ranking
公开(公告)号：US20160305883A1
公开(公告)日：2016-10-20
申请号：US14869968
申请日：2015-09-29
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Dong Li
CPC classification number: G01N21/6458 , G01N21/6428 , G01N2021/6439 , G02B21/06 , G02B21/16 , G02B21/367
Abstract: A method includes: (a) providing spatially-patterned activation radiation to a sample that includes phototransformable labels, where an optical parameter of the spatially-patterned activation radiation varies periodically in space; (b) providing spatially-patterned excitation radiation to the sample, where an optical parameter of the spatially-patterned excitation radiation varies periodically in space, where (a) and (b) create a non-linear fluorescence emission pattern within the sample, the pattern including H modulation harmonics, with H>1. The method includes (c) detecting radiation emitted from the activated and excited labels, (d) storing detected radiation data, and (e) spatially shifting one or both of the spatially-patterned excitation radiation and the spatially-patterned activation radiation with respect to the sample to spatially shift the non-linear fluorescence emission pattern within the sample, and (f) repeating (a)-(e) at least N times, with N>2. Then, a sub-diffraction-limited final image of the sample is generated based on the stored data for the N positions of the non-linear fluorescence emission pattern within the sample.
Abstract translation: 一种方法包括：（a）向包括光变形标签的样品提供空间图案化的激活辐射，其中空间图案化的激活辐射的光学参数在空间中周期性地变化; （b）向样品提供空间图案化的激发辐射，其中空间图案化激发辐射的光学参数在空间中周期性变化，其中（a）和（b）在样品内产生非线性荧光发射图案， 模式包括H调制谐波，H> 1。 该方法包括（c）检测从激活和激发的标签发射的辐射，（d）存储检测到的辐射数据，和（e）空间上移动空间图案化的激发辐射和空间图案化的激活辐射之一或两者相对于 样品空间上移动样品内的非线性荧光发射图案，（f）重复（a） - （e）至少N次，N> 2。 然后，基于样本内的非线性荧光发射图案的N个位置的存储数据生成样本的亚衍射极限最终图像。
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公开(公告)号：US10721441B2
公开(公告)日：2020-07-21
申请号：US16044449
申请日：2018-07-24
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Kai Wang
Abstract: An apparatus includes a light source configured for generating a coherent light beam having a wavelength, λ, a light detector, and beam-forming optics configured for receiving the generated light beam and for generating a plurality of substantially parallel Bessel-like beams directed into a sample in a first direction. Each of the Bessel-like beams has a fixed phase relative to the other Bessel-like beams. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beams and for imaging the received light onto the detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction, where the detector is configured for detecting light received by the imaging optics. A processor configured to generate an image of the sample based on the detected light.
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公开(公告)号：US10509217B2
公开(公告)日：2019-12-17
申请号：US15843633
申请日：2017-12-15
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig
Abstract: A microscope has a light source for generating a light beam having a wavelength, λ, and beam-forming optics configured for receiving the light beam and generating a Bessel-like beam that is directed into a sample. The beam-forming optics include an excitation objective having an axis oriented in a first direction. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beam and for imaging the received light on a detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction. A detector is configured for detecting signal light received by the imaging optics, and an aperture mask is positioned.
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公开(公告)号：US20190199969A1
公开(公告)日：2019-06-27
申请号：US16044449
申请日：2018-07-24
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig , Kai Wang
CPC classification number: H04N7/18 , G02B21/002 , G02B21/06 , G02B27/095
Abstract: An apparatus includes a light source configured for generating a coherent light beam having a wavelength, λ, a light detector, and beam-forming optics configured for receiving the generated light beam and for generating a plurality of substantially parallel Bessel-like beams directed into a sample in a first direction. Each of the Bessel-like beams has a fixed phase relative to the other Bessel-like beams. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beams and for imaging the received light onto the detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction, where the detector is configured for detecting light received by the imaging optics. A processor configured to generate an image of the sample based on the detected light.
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公开(公告)号：US09891421B2
公开(公告)日：2018-02-13
申请号：US14215976
申请日：2014-03-17
Applicant: Howard Hughes Medical Institute
Inventor： Robert E. Betzig
CPC classification number: G02B21/36 , G02B21/0004 , G02B21/002 , G02B21/0096 , G02B21/06 , G02B21/361 , G02B21/365 , G02B27/095
Abstract: A microscope has a light source for generating a light beam having a wavelength, λ, and beam-forming optics configured for receiving the light beam and generating a Bessel-like beam that is directed into a sample. The beam-forming optics include an excitation objective having an axis oriented in a first direction. Imaging optics are configured for receiving light from a position within the sample that is illuminated by the Bessel-like beam and for imaging the received light on a detector. The imaging optics include a detection objective having an axis oriented in a second direction that is non-parallel to the first direction. A detector is configured for detecting signal light received by the imaging optics, and an aperture mask is positioned.
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