WO2019232875A1 - Procédé et dispositif d'imagerie microscopique hyperspectrale chromatographique à large champ de vision fondée sur la focalisation spatiale et temporelle - Google Patents

Procédé et dispositif d'imagerie microscopique hyperspectrale chromatographique à large champ de vision fondée sur la focalisation spatiale et temporelle Download PDF

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WO2019232875A1
WO2019232875A1 PCT/CN2018/094531 CN2018094531W WO2019232875A1 WO 2019232875 A1 WO2019232875 A1 WO 2019232875A1 CN 2018094531 W CN2018094531 W CN 2018094531W WO 2019232875 A1 WO2019232875 A1 WO 2019232875A1
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sample
optical
scanning
pulse laser
ultra
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PCT/CN2018/094531
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Chinese (zh)
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孔令杰
谢浩
张元龙
戴琼海
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清华大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0073—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by tomography, i.e. reconstruction of 3D images from 2D projections

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  • the invention relates to a wide-field tomography hyperspectral microscopic imaging method and device based on space-time focusing, and belongs to the technical field of microspectral imaging and analytical chemistry.
  • Hyperspectral Microscopy has important applications in the field of biomedical research, especially in the fields of clinical disease diagnosis and intraoperative image navigation.
  • the use of hyperspectral microscopy imaging technology to obtain spatially resolvable spectral information can provide information on physiological parameters, morphology, and components of biological tissues for disease diagnosis.
  • non-invasive detection of a variety of cancers has been achieved using hyperspectral microscopy imaging technology.
  • hyperspectral microscopic imaging technology is a technology that obtains higher-dimensional information (ie, spectral information) on the basis of microscopic imaging.
  • the current hyperspectral microscopic imaging technology can be divided into hyperspectral microscopic imaging technology based on ordinary wide-field microscopy, and hyperspectral microscopic imaging technology based on confocal scanning.
  • the former can acquire spectral information in a wide field of view quickly and in parallel, but it is limited to the disadvantages that ordinary wide-field microscopy does not have tomographic capability and is susceptible to signal crosstalk caused by tissue scattering. This technology is only applicable to transparent biological samples.
  • the latter is based on the principle of confocal, to a certain extent, it suppresses the influence of tissue scattering and obtains the ability of axial resolution, but because of the need for point-by-point scanning imaging, the imaging speed is limited.
  • a hyperspectral microscopy imaging technique based on light sheet microscopy has also emerged. Unfortunately, this technique is also not suitable for scattering tissue imaging.
  • non-linear optical microscopy In order to overcome the influence of biological tissue scattering and improve the depth of imaging penetration, non-linear optical microscopy has been introduced into hyperspectral microscopy. Hyperspectral microscopy based on nonlinear optical effects has been developed and widely used in biology. Medical Research. Because ordinary non-linear optical microscopy techniques still use point scanning to overcome the effects of tissue scattering, the speed and flux of imaging will be affected. On the other hand, although surface-excited non-linear optical microscopy eliminates the speed bottleneck caused by point-by-point scanning, the excited signal is severely cross-talked by tissue scattering and is not suitable for scattering tissue spectral imaging. The ordinary non-linear optical microscopy technique using line scanning method can compromise the imaging speed and suppress the influence of scattering. However, compared with the point scanning method, the axial resolution obtained by this method is reduced, which is not an ideal choice.
  • the purpose of the present invention is to overcome the shortcomings of the prior art and provide a wide-field tomography hyperspectral microscopic imaging method and device based on space-time focusing.
  • the invention is suitable for scattering biological tissue imaging, can improve imaging speed and flux, and has high axial resolution.
  • Parameter setting Set the x-axis, y-axis, and z-axis along the horizontal, vertical, and axial directions of the sample, set the ⁇ axis along the direction of the laser spectrum, and the t-axis along the time dimension; set the sample Within the target scanning area XYZ, set the step length of the deflection angle of the galvanometer for scanning along the longitudinal line of the sample, set the axial step of the micro objective lens for scanning along the axial direction of the sample, and set the spectral information according to the size of the target scanning area Acquisition cycle and total scan duration;
  • a non-linear optical effect is used to excite a fluorescent signal on the focus line of step 3).
  • the fluorescent signal is collected and transmitted in the reverse direction through a microscope objective lens, and then the reflected ultra-short pulse laser light is filtered by a filter and confocal optics is used.
  • the slit filters out the stray light caused by the scattering of the sample to obtain the stripe fluorescence of the sample;
  • the obtained bar-shaped emission fluorescence is spectrally expanded by a dispersive element, and the area array detector is used to collect spectral information to obtain (x, ⁇ ) two-dimensional information of the sample; at the same time, according to the set galvanometer deflection angle step size Change the deflection angle of the line scan to obtain (x, ⁇ , y) three-dimensional information of the sample, until the scan traverses the XY target area, and complete the acquisition of spectral information at different positions on the two-dimensional plane of the sample; wherein the detection area of the area array detector The size completely covers the expansion degree of the strip-shaped emission fluorescence, and the area array detector is synchronized with the trigger signal of the galvanometer;
  • the invention also proposes a device according to the above-mentioned wide-field tomography hyperspectral microscopic imaging method based on space-time focusing, which is characterized by comprising an ultra-short pulse laser light source and a beam conversion system, a line-scanning system based on space-time focusing, and optical Microscopy system and filtering and synchronous spectrum confocal detection system; among them,
  • the ultra-short pulse laser light source and a beam conversion system the ultra-short pulse laser light source is used to provide excitation pulse light that generates a non-linear optical signal, and the beam conversion system is used to adjust the excitation pulse light beam size;
  • the space-time focusing-based line scanning system is disposed behind the beam conversion system and includes an optical diffraction element, a lens, and an optical scanning element, and the optical diffraction element and the optical scanning element are respectively placed on an object side of the lens.
  • the optical microscope system is disposed behind the optical scanning element and includes a lens group and a microscope objective lens.
  • the lens group connects the optical scanning element and the microscope objective lens and enters a pupil surface to form a 4f system.
  • the microscope system is used to form a focus line in the sample that is focused in both the space and time dimensions to excite the tissue sample and generate emission fluorescence based on nonlinear optical effects;
  • the filtering and synchronous spectrum confocal detection system is placed in the space-time-focusing-based line-scanning system after an optical scanning element that emits fluorescence collected by the microscopic objective lens and transmitted in the reverse direction, including a filter, a confocal optical narrow
  • the slit and spectrometer are used to select the emitted fluorescent signal of the sample and collect the spectral information; the information acquisition of the spectrometer is synchronized with the trigger signal of the space-time focusing-based line scanning system.
  • the present invention has the following advantages: By adopting a line-scanning technique based on space-time focusing method and proposing a corresponding synchronous spectral confocal detection technique, high axial resolution, low scattered signal crosstalk, and high-speed spectrum can be ensured Information acquisition, the realized wide-field tomography hyperspectral microscopy imaging technology is suitable for deep field wide-field tomography high-spectrum microscopy imaging.
  • the proposed wide-field tomography hyperspectral microscopy imaging technology can be used to obtain (x, y, z, t, ⁇ ) five-dimensional information of deep biological tissues, with a wide field of view, high spatial resolution, High time resolution, high spectral resolution and other advantages.
  • FIG. 1 is a schematic structural diagram of a wide-field tomography hyperspectral microscopic imaging device according to the present invention.
  • Fig. 2 is a schematic diagram of the principle of the present invention.
  • FIG. 3 is a schematic structural diagram of Embodiment 1 of the device of the present invention.
  • FIG. 4 is a schematic structural diagram of a device implementation example 2 of the present invention.
  • Parameter setting Set the x-axis, y-axis, and z-axis along the horizontal, vertical, and axial directions of the sample, set the ⁇ axis along the direction of the laser spectrum (that is, wavelength), and set the t axis along the time dimension. ;
  • Set the target scanning area XYZ in the sample set the step length of the deflection angle of the galvanometer to scan along the longitudinal line of the sample, set the axial step of the microscope objective lens to scan along the axial direction of the sample, and according to the size of the target scanning area Set the spectrum information collection cycle and total scanning time;
  • focus lines are formed in the sample at the same time in two dimensions of space and time by the space-time focusing method (along the x direction, the y direction scan of the focus line can be achieved by changing the deflection angle of the galvanometer) ;
  • a non-linear optical effect is used to excite a fluorescent signal on the focus line of step 3).
  • the fluorescent signal is collected and transmitted in the reverse direction through a microscope objective lens, and then the reflected ultra-short pulse laser light is filtered by a filter and confocal optics is used.
  • the slit filters out the stray light caused by the scattering of the sample to obtain the stripe fluorescence of the sample;
  • the obtained bar-shaped emission fluorescence is spectrally expanded by a dispersive element, and the area array detector is used to collect spectral information to obtain (x, ⁇ ) two-dimensional information of the sample; at the same time, according to the set galvanometer deflection angle step size Change the deflection angle of the line scan to obtain the (x, ⁇ , y) three-dimensional information of the sample until the scan traverses the XY target area to complete the acquisition of the spectral information at different positions on the two-dimensional plane of the sample; of which, the detection area of the area array detector is completely sized Cover the expansion degree of the bar-shaped emission fluorescence, and the area array detector and the trigger signal of the galvanometer are synchronized;
  • the present invention also proposes a wide-field tomography hyperspectral microscopic imaging device according to the above method.
  • the structure is shown in FIG. 1, and includes: an ultra-short pulse laser light source and a beam conversion system, a line-scanning system based on space-time focusing, and optics.
  • Ultra-short pulse laser light source and beam conversion system.
  • Ultra-short pulse laser light source is used to provide excitation pulse light that generates nonlinear optical signals.
  • the beam conversion system is used to adjust the size of the excitation pulse light beam;
  • a line-scanning system based on space-time focusing is placed behind the above-mentioned beam conversion system, and includes an optical diffraction element, a lens, and an optical scanning element.
  • the optical diffraction element and the optical scanning element are respectively placed on the object focal plane and image focal plane of the lens; optical The diffractive element is used to introduce the divergence of the excitation pulse beam, and the optical scanning element is used to introduce the variable deflection angle of the excitation pulse beam;
  • the optical microscope system is placed behind the optical scanning element, and includes a lens group and a microscope objective lens.
  • the lens group connects the optical scanning element and the microscope objective lens and enters the pupil surface, and forms a 4f system for forming simultaneous samples in the sample. Focus lines focused in two dimensions of space and time to excite tissue samples and generate emission fluorescence based on nonlinear optical effects;
  • Filtering and synchronous spectral confocal detection system which is placed in the space-time-focusing-based line-scanning system after the optical scanning element that emits fluorescence collected by the microscopic objective lens and transmitted backward, including a filter, a confocal optical slit and a spectrometer for The emitted fluorescence signal of the sample is selected and the spectral information is collected.
  • the information acquisition of the spectrometer is synchronized with the trigger signal of the line-scanning system based on space-time focusing.
  • a dispersion pre-compensation system is provided before the ultra-short-pulse laser output, which is used to pre-compensate the dispersion accumulated by the ultra-short pulse before it reaches the focus of the microscope objective lens.
  • the line-scan system based on the space-time focusing technology further includes an adaptive optical element placed at the Fourier surface of the optical diffractive element after passing through the lens along the direction of excitation light propagation, which is used to perform spectral phase shaping to further overcome the scattering of biological samples Impact on final imaging.
  • the ultra-short pulse laser light source can select a femtosecond pulse laser light source or a picosecond pulse laser light source according to the output pulse width; the ultra-short pulse laser light source can be adjusted according to whether the output wavelength is adjustable, and a fixed Wavelength ultra-short pulse laser light source or tunable wavelength ultra-short pulse laser light source; the beam conversion system is Galileo telescope system or Kepler telescope system.
  • Ultra-short pulse laser light source and beam conversion system provide any of the non-linear optical signals in the excitation light that generates non-linear optical signals through two-photon absorption fluorescence effect, three-photon absorption fluorescence effect, or two-photon excitation-fluorescence resonance energy transfer effect produce.
  • the optical diffractive element can be a grating, anamorphic mirror, spatial light modulator, or other optical diffractive element;
  • the optical scanning element is a galvanometer, a polygon mirror, or an acousto-optic modulator.
  • the filter is a dichroic mirror, a band-pass filter, a low-pass filter or a long-pass filter.
  • the confocal optical slit is placed on the conjugate surface of the excitation surface of the biological sample, and the width of the confocal optical slit is determined by the design size of the conjugate image of the sample.
  • the spectrometer consists of a dispersive element, a two-dimensional surface detector, and two lenses. The first lens forms an object image relationship between the confocal optical slit and the dispersive element, and the second lens forms an object image relationship between the dispersive element and the two-dimensional surface detector.
  • Dispersion elements can be prisms, gratings or other dispersive elements.
  • Two-dimensional surface detectors use charge-coupled elements (CCD), electron multiplying charge-coupled elements (EMCCD), or scientific-grade complementary metal-oxide-semiconductor devices (sCMOS).
  • CCD charge-coupled elements
  • EMCD electron multiplying charge-coupled elements
  • sCMOS scientific-grade complementary metal-oxide-semiconductor devices
  • FIG. 2 a schematic diagram of the principle of the present invention is shown.
  • the use of non-linear optical effects combined with space-time focusing technology can generate a focus line (x direction) with high axial resolution on a biological sample.
  • the excited fluorescence spectrum signal is detected by confocal detection Imaging on the two-dimensional photoelectric detection surface (Dx, Dy), where the Dy direction is the lambda dimension of the spectrum.
  • Dx, Dy two-dimensional photoelectric detection surface
  • (x, ⁇ , y, z) four-dimensional information of the sample can be obtained; further, using this technology's High-speed spectral microscopic imaging capability enables time-delayed information collection (that is, collecting (x, ⁇ , y, z) four-dimensional information of samples at different times) to obtain (x, ⁇ , y, z, t) five-dimensional information of samples.
  • the wide-field tomographic hyperspectral micro-imaging method and device provided by the present invention have the advantages of wide field of view, high spatial resolution, high temporal resolution, high spectral resolution, etc., and can be used for biological dynamic process research, Provides a wealth of information on the basis of disease diagnosis.
  • FIG. 3 includes an ultra-short pulse laser light source and a beam conversion system, a line-scanning system based on space-time focus, an optical microscope system, and filtering.
  • a biological sample is placed on a sample stage 319.
  • the ultra-short pulse laser light source and the ultra-short pulse laser light source 301 in the beam conversion system use a femtosecond laser (such as the Coherent Chameleon Discovery series), and the beam conversion system uses a Kepler telescope system composed of a lens 302 and a cylindrical lens 303 ( Is a 4f system); a line-scan system based on space-time focusing includes a transmission grating 304, a lens 305, and a scanning galvanometer 307; an optical microscope system includes two lenses 308, 309, and a microscope objective 310; a filtering and synchronous spectral confocal detection system It includes a dichroic mirror 306, a low-pass filter 311, a lens 312, a confocal optical slit 313 (the width of the optical slit is determined by the design size of the conjugate image of the biological sample), and two lenses 314, 316 and a reflection grating 315 And two-dimensional surface detector (using sCMOS or
  • the lens 302 and the cylindrical lens 303 form a 4f system to expand the beam
  • the transmission grating 304 is placed at the image surface of the cylindrical lens 303
  • the lens 305 forms the transmission grating 304 at the scanning galvanometer 307.
  • 308 and 309 form a 4f system so that the scanning galvanometer 307 and the entrance pupil plane of the micro objective 310 are conjugated
  • the lens 312 and lens 308 form a 4f system so that the object plane is imaged at the confocal optical slit 313.
  • FIG. 3 also illustrates a computer 318 for controlling the deflection angle of the scanning galvanometer 307, and performing conventional image reconstruction and data processing on the spectral information collected by the two-dimensional surface detector 317.
  • the laser beam emitted by the ultra-short pulse laser light source 301 is expanded by the lens 302 and the cylindrical lens 303 (changing the diameter of the laser beam) and then incident on the transmission grating 304.
  • the ultra-short pulse beam is generated by the transmission grating 304.
  • Role divergence (the purpose is to make the introduced divergent beam fill the back focal plane of the objective lens after the subsequent optical elements), collimated by the lens 305 and projected onto the scanning galvanometer 307 through the dichroic mirror 306 and introducing a variable deflection angle (The deflection angle is driven by the galvanometer driving voltage, and the deflection angle is set according to the scanning area.) Finally, a focus line is generated on the focal surface of the objective lens in the biological sample through the lenses 308 and 309 and the micro objective lens 310.
  • the optical signal generated by the non-linear optical effect is collected by the micro objective lens 310 and transmitted in the reverse direction, passes through the lenses 309 and 308 and the scanning galvanometer 307 in order, and is reflected by the dichroic mirror 306. After that, the signal beam passes through the low-pass filter 311, the lens 312, and the confocal optical slit 313 in order, and finally enters the spectrometer for signal collection.
  • the line scanning trigger signal of the scanning galvanometer 307 is synchronized with the frame trigger signal of the two-dimensional area detector 317.
  • (x, ⁇ , y, z) By moving the microscope objective 310 for axial scanning, (x, ⁇ , y, z) four-dimensional information of the sample can be obtained. If delayed information collection is performed, (x, ⁇ , y, z, t) five-dimensional information of the sample can be obtained.
  • the following describes in detail the wide-field tomography hyperspectral microscopic imaging device of this embodiment with reference to FIG. 4.
  • the difference between this embodiment and Embodiment 1 is that an adaptive optical element is added.
  • the device of this embodiment includes an ultra-short pulse laser light source and a beam conversion system, a space-time focusing-based line scanning system, an optical microscopy system, and a filtering and synchronous spectral confocal detection system.
  • a biological sample is placed on the sample stage 424;
  • the pulsed laser light source and the ultra-short pulsed laser light source 401 in the beam conversion system use a femtosecond laser (such as the Coherent Chameleon Discovery series), and the beam conversion system uses a Kepler telescope system composed of a lens 402 and a cylindrical lens 403 (for a 4f system) ;
  • Space-time focusing-based line scan system includes a transmission grating 404, five lenses 405, 407, 408, 410, 411, and an adaptive optical element 406 disposed between the lenses 405 and 407 (a spatial light modulator is used in this embodiment) , Scanning galvanometer 412;
  • optical microscope system includes two lenses 413, 414, microscope objective lens 415;
  • filtering and synchronous spectrum confocal detection system includes dichroic mirror 409, low-pass filter 416, ordinary lens 417, confocal optics Slot 418 (the width of the optical slit is determined by the design size of
  • the lens 402 and the cylindrical lens 403 form a 4f system to expand the beam
  • the transmission grating 404 is placed at the image surface of the cylindrical lens 403
  • the lens 405 images the transmission grating 404 at the spatial light modulator 406.
  • the lenses 407 and 408, the lenses 410 and 411, and the lenses 413 and 414 respectively constitute three groups of 4f systems connected in series, so that the spatial light modulator 406 is conjugated with the entrance pupil plane of the scanning galvanometer 412 and the micro objective lens 415, and the lenses 417 and 410 constitute
  • the 4f system makes the object surface image at the confocal optical slit 418, the low-pass filter 416 is placed immediately before the lens 417, the lens 419 images the confocal optical slit 418 at the reflection grating 420, and the lens 421 images the reflection grating 420 at Two-dimensional surface detector 422.
  • FIG. 4 also illustrates a computer 423 for controlling the deflection angle of the scanning galvanometer 412, and performing conventional image reconstruction and data processing on the spectral information collected by the two-dimensional surface detector 422.
  • the laser beam emitted by the ultra-short pulse laser light source 401 is expanded by the lens 402 and the cylindrical lens 403 and then enters the transmission grating 404.
  • the ultra-short pulse beam is scattered and passes through the lens.
  • 405 is collimated and projected to spatial light modulator 406 for spectral phase shaping to further overcome the influence of tissue scattering, and then passes through lens 407, lens 408, dichroic mirror 409, lens 410, and lens 411 on scanning galvanometer 412.
  • the variable deflection angle finally generates a focus line on the focal surface of the objective lens in the biological sample through the lenses 413, 414 and the micro objective lens 415.
  • the fluorescence signal generated by the nonlinear optical effect is collected by the micro objective lens 415 and transmitted in the reverse direction, passes through the lens 414, the lens 413, the scanning galvanometer 412, the lens 411, and the lens 410, and is reflected by the dichroic mirror 409. After that, the signal beam passes through the low-pass filter 416, the lens 417, and the confocal optical slit 418 in order, and finally enters the spectrometer for signal collection.
  • the line trigger signal of the scanning galvanometer 412 is synchronized with the frame trigger signal of the two-dimensional surface detector 422; the spatial light modulator 406 uses adaptive optics to measure the wavefront distortion and applies a compensated wavefront (in the spectrum Perform before imaging).
  • (x, ⁇ , y) three-dimensional information of a biological sample can be obtained.
  • (x, ⁇ , y, z) four-dimensional information of a biological sample can be obtained. If time-delayed information collection is performed (that is, multiple acquisitions), (x, ⁇ , y, z, t) five-dimensional information of the biological sample can be obtained.
  • the required excitation light power may be greater than the light damage threshold of the spatial light modulator
  • the present invention combines line-time focusing technology to perform line scanning technology, and proposes a corresponding synchronous spectral confocal detection technology, which ensures high axial resolution and low scattering signal crosstalk, and is suitable for deep tissue tomography microscopy imaging. ; Improve the speed of obtaining spectral information, and can achieve high-speed spectral microscopy imaging in a wide field of view.

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Abstract

La présente invention se rapporte au domaine technique de l'imagerie spectrale microscopique et de la chimie analytique, et concerne un procédé et un dispositif d'imagerie microscopique hyperspectrale chromatographique à large champ de vision fondée sur la focalisation spatiale et temporelle. Le procédé consiste à : utiliser une source laser à impulsions ultracourtes pour produire un laser à impulsions ultracourtes; produire une ligne focale dans un échantillon par focalisation spatiale et temporelle; collecter une lumière fluorescente excitée et filtrer la lumière parasite en utilisant une fente optique confocale; collecter des informations relatives au spectre de fluorescence pour terminer l'acquisition d'informations relatives au spectre (x, λ) de l'échantillon; et enfin, acquérir des informations en cinq dimensions (x, λ, y, z, t) concernant l'échantillon par balayage spatial tridimensionnel et balayage retardé. Le dispositif comprend un système de conversion de faisceau et de source laser à impulsions ultracourtes, un système de balayage de ligne fondé sur la focalisation spatiale et temporelle, un système microscopique optique et un système de détection confocale qui filtre et synchronise les spectres; l'acquisition d'informations relatives aux spectres dans le système de détection confocale qui filtre et synchronise les spectres est synchronisée avec un signal de déclenchement de balayage de ligne dans le système de balayage de ligne qui combine une technologie de focalisation spatiale et temporelle. La présente invention présente les avantages suivants : un large champ de vision, une résolution spatiale élevée, une résolution temporelle élevée, une résolution spectrale élevée et équivalent.
PCT/CN2018/094531 2018-06-08 2018-07-04 Procédé et dispositif d'imagerie microscopique hyperspectrale chromatographique à large champ de vision fondée sur la focalisation spatiale et temporelle WO2019232875A1 (fr)

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Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107290286A (zh) * 2016-04-12 2017-10-24 北京世纪桑尼科技有限公司 一种可用于光谱分析的高速扫描共聚焦成像系统
CN109799602A (zh) * 2018-12-24 2019-05-24 清华大学 一种基于线扫描时空聚焦的光显微成像装置及方法
CN110031830B (zh) * 2019-04-29 2020-11-03 清华大学深圳研究生院 一种基于激光线扫描成像的测距方法
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US11333870B2 (en) * 2019-12-04 2022-05-17 National Taiwan University Large-angle optical raster scanning system for deep tissue imaging
DE102020108117B4 (de) * 2020-03-24 2023-06-15 Evident Technology Center Europe Gmbh Mikroskop und Verfahren zum Betreiben eines Mikroskops
CN112903640B (zh) * 2021-01-19 2023-01-03 雷振东 光子反冲成像共聚焦探测系统及方法
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CN113558576B (zh) * 2021-07-27 2023-12-15 苏州微景医学科技有限公司 激光扫描成像方法、系统及存储介质
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CN113702288B (zh) * 2021-08-18 2022-07-01 北京大学 一种双模态显微成像系统及其成像方法
CN113984715A (zh) * 2021-10-28 2022-01-28 上海盛晃光学技术有限公司 相干断层扫描装置及方法
CN114326099B (zh) * 2021-12-29 2024-04-02 武汉大学 一维高速、高分辨率成像系统及基于该系统的实时熔池监测方法
CN114326100A (zh) * 2021-12-29 2022-04-12 武汉大学 一种二维高速、高分辨率成像系统及基于该系统的实时熔池监测方法
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060256348A1 (en) * 2005-03-25 2006-11-16 Fuji Photo Film Co., Ltd. Optical tomography apparatus
CN107144955A (zh) * 2017-05-15 2017-09-08 清华大学 基于线扫描时空聚焦的结构光显微成像系统
CN107966802A (zh) * 2017-12-26 2018-04-27 清华大学 基于相机阵列的超光谱光片光场显微成像系统及方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8848199B2 (en) * 2007-07-10 2014-09-30 Massachusetts Institute Of Technology Tomographic phase microscopy
CN101401722B (zh) * 2008-11-07 2012-07-25 上海奥通激光技术有限公司 一种多模式共聚焦成像方法及其装置
US8570405B2 (en) * 2010-08-11 2013-10-29 Inview Technology Corporation Determining light level variation in compressive imaging by injecting calibration patterns into pattern sequence

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060256348A1 (en) * 2005-03-25 2006-11-16 Fuji Photo Film Co., Ltd. Optical tomography apparatus
CN107144955A (zh) * 2017-05-15 2017-09-08 清华大学 基于线扫描时空聚焦的结构光显微成像系统
CN107966802A (zh) * 2017-12-26 2018-04-27 清华大学 基于相机阵列的超光谱光片光场显微成像系统及方法

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