WO2020062609A1 - 用于sted光学显微镜的照明系统及sted光学显微镜 - Google Patents

用于sted光学显微镜的照明系统及sted光学显微镜 Download PDF

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Publication number
WO2020062609A1
WO2020062609A1 PCT/CN2018/121206 CN2018121206W WO2020062609A1 WO 2020062609 A1 WO2020062609 A1 WO 2020062609A1 CN 2018121206 W CN2018121206 W CN 2018121206W WO 2020062609 A1 WO2020062609 A1 WO 2020062609A1
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Prior art keywords
light
light beam
illumination
sted
incident
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PCT/CN2018/121206
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English (en)
French (fr)
Chinese (zh)
Inventor
袁景和
于建强
方晓红
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Institute of Chemistry CAS
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Institute of Chemistry CAS
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Priority to US17/043,615 priority Critical patent/US11726309B2/en
Priority to JP2020551913A priority patent/JP7073523B2/ja
Priority to EP18934542.4A priority patent/EP3757650B1/en
Publication of WO2020062609A1 publication Critical patent/WO2020062609A1/zh
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/58—Optics for apodization or superresolution; Optical synthetic aperture systems

Definitions

  • the invention relates to the field of microscopic imaging technology, in particular to an illumination system for a STED optical microscope and a STED optical microscope.
  • STED Stimulated Emission Depletion-STED
  • STED uses a beam of STED light to form an empty shell-shaped light spot, and converts the fluorescent molecules around the excitation light diffraction spot to a non-radiative state through depletion of stimulated radiation. Spatial resolution at 50nm. Due to the use of a plenoptic setting, the image acquisition time is the same as a traditional confocal microscope, and there are no special requirements for sample preparation, so real-time imaging and dynamic tracking of subcellular structures in living cells can be achieved.
  • the object of the present invention is to solve at least one of the above-mentioned defects and deficiencies, and the object is achieved by the following technical solutions.
  • the invention provides an illumination system for a STED optical microscope, which includes an illumination light source and an illumination light path composed of optical elements.
  • the light beam emitted by the illumination light source passes through the illumination light path and is focused and irradiated onto a sample surface to excite the fluorescence in the sample.
  • the substance emits fluorescence, and the illumination light path includes a first filter, a second filter, a polarization beam splitter, a first 1/4 wave plate, a first dichroic element, and an optical path sequentially arranged along a transmission direction of the optical path.
  • the light beam emitted by the illumination light source passes through the first filter and the second filter to obtain a certain wavelength.
  • a first light beam and a second light beam are respectively split by the polarization beam splitter to form linearly polarized light, and are sequentially incident on the first 1/4 wave plate and
  • the first light beam forms circularly polarized light after passing through the first 1/4 wave plate, and after being reflected by the first dichroic element, passes through the first Linearly polarized light is formed after a 1/4 wave plate, and then After being transmitted by the polarizing beam splitter, reflecting by the second dichroic element, and converted by the second quarter-wave plate into circularly polarized light, the microscope objective lens that is incident on the microscopic imaging system converges in the display
  • a first light spot is formed at the focal surface of the micro objective lens; the second light beam passes through the first quarter-wave plate to form circularly polarized light, is transmitted through the first dichroic element, and enters the optical path A delay unit and the phase plate, after the second light beam is reflected by the phase plate, it is sequentially emitted through
  • the first light beam is excitation light
  • the first light spot is a solid light spot
  • the second light beam is lossy light with respect to the first light beam
  • the second light spot is a hollow light spot.
  • the first filter is a neutral filter for adjusting the intensity of the total laser light emitted by the illumination light source; and the second filter is a dual band-pass filter for Filtering out the first beam and the second beam with a certain wavelength, and adjusting the intensity of the first beam and the intensity of the second beam, the first filter and the second filter
  • the light sheet is arranged coaxially along the optical path.
  • the first 1/4 wave plate can convert the incident first and second light beams from linearly polarized light to circularly polarized light, and can also convert the incident first and second light beams from Circularly polarized light is converted into linearly polarized light; the second quarter-wave plate is capable of converting linearly polarized light into circularly polarized light.
  • the first dichroic element is a selective transmission dielectric film, the dielectric film is plated on the incident end of the optical path delay unit, and the dielectric film can reflect the incident first light beam. And the incident second light beam can be transmitted; the second dichroic element is a dichroic sheet, and the dichroic sheet can make the incident first light beam and the second light beam both Reflected and capable of transmitting the fluorescence emitted by the sample.
  • the optical path delay unit can cause an optical delay to the second light beam.
  • phase plate is a reflective phase plate, which is disposed at the far end of the optical path delay unit, and the second light beam enters the optical path delay unit and is incident on the phase plate. Returning to the incident end of the optical path delay unit along the original optical path, and the phase plate can modulate the wavefront of the incident second light beam.
  • a beam expander is provided between the illumination light source and the first filter to expand and shape the light beam emitted by the illumination light source.
  • the present invention also provides an illumination system for the STED optical microscope.
  • the STED optical microscope further includes a microscopic imaging system and a fluorescence detection system.
  • the microscopic imaging system includes a microscopic objective lens, and the illumination light source emits light. After passing through the illumination light path of the illumination system, the light beam is divided into two coaxial first light beams and a second light beam, and the first light beam and the second light beam are respectively focused by the microscope objective lens and then irradiated onto the sample to be excited.
  • the fluorescent substance in the sample emits fluorescence, and the fluorescence is collected by the microscope objective lens to enter the fluorescence detection system for detection, and the optical axis of the first beam and the optical axis of the second beam are both The optical axis of the detection optical path of the fluorescence detection system is coaxial.
  • a third filter for filtering the fluorescence is provided between the emission end of the microscopic imaging system and the fluorescence detection system.
  • the illumination system for the STED optical microscope provided by the present invention is integrated and designed to avoid the physical adjustment of the mutual geometrical relationship of each unit device and the inherent temperature and vibration instability of the mechanical adjustment mechanism, which makes the STED instruments and equipment Capable of long-term reliable operation in a variety of environments.
  • the present invention can filter the excitation light and loss light of appropriate wavelengths from the laser light source through optical cold working and optical coating.
  • the optical coating also realizes the adjustment of the relative strength of the two, and the neutral filter achieves the total light intensity. Regulation.
  • the present invention uses a combination of a polarizing beam splitter and a wave plate to make the optical path compact, which is beneficial to the use of fewer optical elements to realize the steering of the light beam and the polarization state of the light beam, and enables each main optical element to be bonded through the end face.
  • the methods are combined into an integrated integrated module, which reduces the volume of the device while making the device insensitive to temperature and environmental vibration, and improves reliability.
  • phase plate is processed on the optical path delay unit through a binary optical processing process, and reflective phase adjustment is performed on the loss light, which can generate the critical and concentric excitation light in the STED technology with a certain delay.
  • Phase-modulated light beam (lossy light is collected by the objective lens to form an empty shell-shaped light spot).
  • FIG. 1 is a schematic structural diagram of an illumination system for a STED optical microscope according to an embodiment of the present invention
  • 2 is a confocal imaging image obtained by irradiating a 40 nm fluorescent microsphere sample according to an existing confocal microscope;
  • FIG. 3 is a STED image obtained by irradiating a 40 nm fluorescent microsphere sample with the illumination system for a STED optical microscope provided in an embodiment of the present invention in the same region of FIG. 2;
  • FIG. 3 is a STED image obtained by irradiating a 40 nm fluorescent microsphere sample with the illumination system for a STED optical microscope provided in an embodiment of the present invention in the same region of FIG. 2;
  • FIG. 1 is a schematic structural diagram of an illumination system for a STED optical microscope according to an embodiment of the present invention.
  • the illumination system is suitable for a STED optical microscope.
  • the STED optical microscope includes an illumination system 10, a microscopic imaging system 20, and a fluorescence detection system 30.
  • the light beam emitted by the illumination system 10 is focused by the microscope objective lens of the microscopic imaging system 20 and irradiated onto the sample surface to excite the fluorescent substance in the sample to emit fluorescence.
  • the fluorescence of the sample is converged by the microscope objective lens and incident on the fluorescence detection system 30 Check it out.
  • the illumination system for a STED optical microscope includes an illumination light source and an illumination light path composed of optical elements.
  • the illumination light source is a laser 1 capable of emitting multi-wavelength laser light or multi-laser light. combination.
  • the laser beam emitted by the laser 1 is focused on the sample surface after passing through the illumination light path.
  • the illumination light path includes a first filter 21, a second filter 22, a polarization beam splitter 3, and a first 1/4, which are sequentially arranged along the transmission direction of the optical path.
  • the wave plate 41, the first dichroic element 51, the optical path delay unit 6, the phase plate 7, the second dichroic element 52, and the second 1/4 wave plate 42, the light beam emitted from the illumination light source 1 passes through the first
  • the filter 21 and the second filter 22 are filtered to obtain two light beams, a first beam 101 and a second beam 102.
  • the first beam 101 and the second beam 102 are separated by a polarization beam splitter 3 and incident on the first 1
  • the / 4 wave plate 41 generates circularly polarized light and enters the first dichroic element 51.
  • the first light beam 101 incident on the first dichroic element 51 is reflected by the first 1/4 wave plate 41 to generate linearly polarized light, transmitted by the polarizing beam splitter 3, reflected by the second dichroic element 52, and the first
  • the second quarter-wave plate 42 is converted into circularly polarized light and incident on the microscope objective lens to form a first light spot at the focal surface of the objective lens;
  • the second light beam 102 incident on the first dichroic element 51 is incident after transmission An optical delay is generated until the optical path delay unit 6, the second light beam 102 is reflected by a phase plate 7 disposed at the far end of the optical path delay unit 6, and is sequentially emitted through the optical path delay unit 6, the first dichroic element 51 is transmitted,
  • the first quarter-wave plate 41 generates linearly polarized light
  • the transmission of the polarizing beam splitter 3, the reflection from the second dichroic element 52, and the second quarter-wave plate 42 is converted into circularly polarized light and is also incident on the microscope objective lens for convergence
  • the first light beam 101 is excitation light, which is used to excite fluorescence and image the sample.
  • the second light beam 102 is loss light relative to the first light beam 101, which is used to suppress fluorescence, and de-excites fluorescence in the peripheral region of the first spot.
  • the fluorescent substance in the emission state makes the peripheral region no longer generate fluorescence.
  • the first filter 21 is a neutral filter.
  • the first filter 21 can filter the light beam and adjust the intensity of the total laser light emitted by the illumination light source 1.
  • the second filter 22 is a dual band-pass filter. It is used to filter out the first light beam 101 and the second light beam 102 with suitable wavelengths, and adjust the intensity of the two light beams.
  • the second filter 22 uses an optical coating to regulate the relative intensity of both the excitation light and the loss light.
  • a beam expanding mirror 8 is provided between the laser 1 and the first filter 21.
  • the beam expanding mirror 8 can expand and shape the laser beam.
  • the first filter 21 and the second filter 22 are coaxially arranged along the optical path and coincide with the optical axis of the laser 1; the axis line of the beam expander 8 also coincides with the optical axis of the laser 1.
  • the polarization beam splitter 3 can separate incident light into two linearly polarized lights whose polarization directions are perpendicular to each other, and respectively reflect and transmit the same.
  • the first light beam 101 and the second light beam 102 pass through the polarization beam splitter 3 to form linearly polarized light.
  • the P-polarized light beam is discarded after transmission, and the S-polarized light beam is incident on the first quarter-wave plate 41 and the first one in turn Dichroic element 51.
  • the first 1/4 wave plate 41 and the second 1/4 wave plate 42 are used to adjust the polarization state of the light beam.
  • the first 1/4 wave plate 41 can make the incident first light beam 101 and the second light beam 102 be linearly polarized light. Converting to circularly polarized light can also convert the incident first and second light beams 101 and 102 from circularly polarized light to linearly polarized light; the second quarter-wave plate can make the first and second light beams 101 and 102 from linear Polarized light is converted into circularly polarized light.
  • the first dichroic element 51 is a selectively transmissive dielectric film capable of reflecting the incident first light beam 101 and transmitting the second light beam 102.
  • the second dichroic element 52 is a selectively transmitting dichroic sheet, which can reflect both the incident first light beam 101 and the second light beam 102 and transmit the fluorescence emitted by the sample.
  • plating the dielectric film of the first dichroic element 51 on the surface of the incident end of the optical path delay unit 6 can not only reduce the volume of the entire optical path system, make the arrangement of the optical elements more reasonable and compact, but also reduce the temperature. , Vibration and other environmental factors on the microscope.
  • the optical path delay unit 6 is optical glass whose two ends are strictly parallel, and its length can be designed according to actual needs, so that it can generate a first light beam 101 and a second light beam 102 with appropriate delays.
  • the phase plate 7 is a reflective phase plate, which is disposed at the far end of the optical path delay unit 6 and is perpendicular to the optical axis of the optical path delay unit 6 (the center line of the phase plate 7 coincides with the optical axis of the optical path delay unit 6), It is used to modulate the loss light wavefront and optical path, and produces an empty shell-shaped focal spot on the focal surface of the microscope objective lens.
  • the reflective phase adjustment of the lossy light on the phase plate 7 can generate the phase-modulated beam that is the key in STED technology and has a certain delay that is strictly concentric with the excitation light (the lossy light is condensed by the objective lens) After forming an empty shell-shaped light spot).
  • the optical path delay unit 6 and the phase plate 7 may be an integrated structure or a split structure.
  • the optical path delay unit 6 and the phase plate 7 are provided as an integrated structure, and the phase plate 7 is processed on the surface of the exit end of the optical path delay unit 6.
  • the first light beam 101 is converted into circularly polarized light by the first 1/4 wave plate 41, and is reflected by the first dichroic element 51, and then linearly polarized light is formed after passing through the first 1/4 wave plate 41, and is incident on The polarization beam splitter 3 transmits.
  • the second light beam 102 is converted into circularly polarized light by the first 1/4 wave plate 41, and then transmitted by the first dichroic element 51 and enters the optical path delay unit 6 to generate an optical delay, and is incident on the optical path delay set.
  • the second light beam 102 is subjected to wavefront modulation reflection by the phase plate 7 and then returns to the incident end of the optical path delay unit 6 along the original optical path, and is transmitted through the first dichroic element 51 to
  • the first quarter-wave plate 41 is converted into linearly polarized light, and then transmitted through the polarization beam splitter 3, and then re-combined with the first beam 101 at the polarization beam splitter 3. Since the first beam 101 does not pass through the optical path delay unit 6 and the second beam 102 passes through the optical path delay unit 6, a fixed pulse time delay is generated between the two, and the first beam 101 and the second beam 102 after recombination Coaxial.
  • the recombined first beam 101 and second beam 102 are respectively reflected by the second dichroic element 52 and then passed through the second quarter-wave plate 42 to convert linearly polarized light into circularly polarized light, and then enter the microscopic imaging
  • the focusing objective lens condenses and irradiates the sample to form a concentric light spot.
  • the first light beam 101 is focused and irradiated on the sample to form a solid spot;
  • the second light beam 102 is focused and irradiated on the sample to form a hollow spot.
  • the first light beam 101 and the second light beam 102 have different wavelengths.
  • the surface of the sample excites fluorescence.
  • the solid light spot overlaps the hollow light spot.
  • the solid light spot excites the fluorescent substance on the sample to emit fluorescence, and the hollow The light spot suppresses the fluorescence emitted by the periphery of the fluorescent substance, so that only the middle point that is smaller than the diffraction limit emits fluorescence and is observed.
  • the excited fluorescence is filtered by the third filter 9 and received by the fluorescence detection system 30 for detection.
  • the first quarter-wave plate 41 converts the light beam reflected by the polarization beam splitter 3 from linearly polarized light to circularly polarized light, and converts the second incident circularly polarized light into linearly polarized light and then enters the polarization beam splitter 3 to occur. Transmission to achieve smooth transmission of the light path.
  • the second quarter-wave plate 42 By using the second quarter-wave plate 42 to set the excitation light entering the microscopic imaging system to circularly polarized light, a higher fluorescence excitation efficiency can be obtained and a better hollow focus spot can be obtained.
  • the polarization beam splitter 3 is connected to the first quarter-wave plate 41, and the first dichroic element 51 is plated on the surface of the optical path delay unit 6 to regulate the polarization properties of the excitation light and the loss light.
  • the optical path transmission path; the second dichroic element 52 is connected to the second quarter-wave plate 42 to adjust the polarization state of the excitation light and the loss light after recombination, and change the light propagation direction.
  • the third filter 9 is a fluorescent band-pass filter, which is disposed between the microscopic imaging system 20 and the fluorescence detection system 30.
  • the fluorescence emitted by the sample is filtered by the third filter 9 to remove light other than fluorescence (including scattering).
  • the third filter 9 is disposed on one side of the second dichroic element 52, and the second dichroic element 52 is provided as a lens capable of transmitting fluorescence, which can ensure the fluorescence emitted by the sample. Direct light enters the fluorescence detection system 30 for detection.
  • FIG. 2 shows an image obtained by performing a confocal imaging test on a fluorescent microsphere sample with a diameter of 40 nm using a conventional confocal microscope
  • FIG. 3 shows a fluorescent microsphere sample with a diameter of 40 nm using the above-mentioned STED optical microscope.
  • STED Stimulated Radiation Depletion
  • the illumination system for a STED optical microscope provided by the present invention can greatly improve the resolution of imaging and obtain the effect of super-resolution imaging.
  • the invention also provides a STED optical microscope including the illumination system described above.
  • the STED optical microscope further includes a microscopic imaging system 20 and a fluorescence detection system 30.
  • the microscopic imaging system 20 includes the microscopic objective lens.
  • the light beam emitted by the illumination light source is divided into two coaxial first beams by the illumination light path of the illumination system. 101 and second light beam 102, the first light beam 101 and the second light beam 102 are respectively converged by a microscope objective lens and irradiated onto a sample to excite a fluorescent substance in the sample to emit fluorescence.
  • the fluorescence is condensed by the microscope objective lens and enters a fluorescence detection system 30 for detection .
  • the optical axes of the detection light paths of the first light beam 101, the second light beam 102, and the fluorescence detection system 30 are coaxial, and the focal plane of the micro objective lens and the light of the detection light paths of the first light beam 101, the second light beam 102, and the fluorescence detection system are coaxial.
  • the axes are all vertical.
  • the illumination system for the STED optical microscope provided by the present invention adopts an integrated and integrated optical module design, can realize coaxial input and output of excitation light, loss light, and confocal detection optical path, and realize excitation light spot and STED hollow shell light spot.
  • first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply these entities or operations There is any such actual relationship or order among them.

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
PCT/CN2018/121206 2018-09-26 2018-12-14 用于sted光学显微镜的照明系统及sted光学显微镜 Ceased WO2020062609A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US17/043,615 US11726309B2 (en) 2018-09-26 2018-12-14 Illumination system for STED optical microscope and STED optical microscope
JP2020551913A JP7073523B2 (ja) 2018-09-26 2018-12-14 Ｓｔｅｄ光学顕微鏡に用いる照明システム及びｓｔｅｄ光学顕微鏡
EP18934542.4A EP3757650B1 (en) 2018-09-26 2018-12-14 Illumination system for sted optical microscope and sted optical microscope

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CN201811125387.8A CN108957720B (zh) 2018-09-26 2018-09-26 受激辐射损耗光学显微镜及其照明系统
CN201811125387.8 2018-09-26

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US (1) US11726309B2 (enExample)
EP (1) EP3757650B1 (enExample)
JP (1) JP7073523B2 (enExample)
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WO (1) WO2020062609A1 (enExample)

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CN115508994A (zh) * 2022-09-14 2022-12-23 郑州思昆生物工程有限公司 一种显微成像系统
CN115728869A (zh) * 2022-11-15 2023-03-03 昂纳信息技术（深圳）有限公司 一种可调谐光滤波器

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CN108957720B (zh) 2018-09-26 2019-12-10 中国科学院化学研究所 受激辐射损耗光学显微镜及其照明系统
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US11271747B2 (en) * 2019-09-16 2022-03-08 Lawrence Livermore National Security, Llc Optical authentication of images
CN113866973B (zh) * 2021-10-12 2023-10-03 桂林电子科技大学 一种基于多阶光纤模式复用的光纤sted显微镜
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CN114706208B (zh) * 2022-02-18 2023-01-17 中国科学院化学研究所 受激辐射损耗光学显微镜及其显微成像系统
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103257130A (zh) * 2013-05-31 2013-08-21 中国科学院苏州生物医学工程技术研究所 受激辐射损耗显微成像系统
CN103616330A (zh) * 2013-11-18 2014-03-05 中国科学院化学研究所 基于超连续产生的宽带激光光源激发的超分辨sted显微成像系统
CN106841149A (zh) * 2017-03-17 2017-06-13 王富 受激辐射损耗显微方法及显微装置
WO2017174100A1 (de) * 2016-04-08 2017-10-12 Universität Heidelberg Parallelisierung des sted-mikroskopieverfahrens
CN108957720A (zh) * 2018-09-26 2018-12-07 中国科学院化学研究所 用于sted光学显微镜的照明系统及sted光学显微镜

Family Cites Families (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003262798A (ja) 2002-03-12 2003-09-19 Olympus Optical Co Ltd 顕微鏡
JP4334835B2 (ja) 2002-08-28 2009-09-30 独立行政法人科学技術振興機構 顕微鏡
DE102007025688A1 (de) 2007-06-01 2008-12-11 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Wellenlängen- oder polarisationssensitiver optischer Aufbau und dessen Verwendung
DE102010060121C5 (de) * 2010-10-22 2023-09-28 Leica Microsystems Cms Gmbh SPIM-Mikroskop mit sequenziellem Lightsheet
JP5984351B2 (ja) 2011-09-14 2016-09-06 キヤノン株式会社 計測装置
US9721326B2 (en) * 2011-11-08 2017-08-01 UNIVERSITé LAVAL Method and system for improving resolution in laser imaging microscopy
CN102540439B (zh) 2011-12-30 2013-10-30 哈尔滨工业大学 基于反射式液晶空间光调制器的共焦轴向扫描装置及共焦轴向扫描方法
JP5988643B2 (ja) 2012-03-26 2016-09-07 キヤノン株式会社 計測装置、計測方法及び光学部品の製造方法
GB201217171D0 (en) 2012-08-23 2012-11-07 Isis Innovation Stimulated emission depletion microscopy
EP3957983B1 (en) * 2013-06-03 2023-12-13 LUMICKS DSM Holding B.V. Method and system for imaging a molecular strand
JP6613552B2 (ja) 2013-09-09 2019-12-04 株式会社ニコン 超解像観察装置及び超解像観察方法
US10247672B2 (en) * 2014-09-29 2019-04-02 Howard Hughes Medical Institute Non-linear structured illumination microscopy
DE102016211374A1 (de) * 2016-06-24 2017-12-28 Carl Zeiss Microscopy Gmbh Mikroskopieverfahren unter Nutzung zeitlicher Fokusmodulation und Mikroskop
US9880377B1 (en) * 2016-09-09 2018-01-30 Photonicsys Ltd. Multiple wavelengths real time phase shift interference microscopy
CN107329245B (zh) * 2017-07-06 2023-05-05 中国科学院西安光学精密机械研究所 基于径向偏振调制的干涉式结构光照明显微镜系统与方法
CN108121059B (zh) * 2017-11-18 2022-01-28 苏州国科医工科技发展(集团)有限公司 一种基于结构光照明的sted并行显微成像系统

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103257130A (zh) * 2013-05-31 2013-08-21 中国科学院苏州生物医学工程技术研究所 受激辐射损耗显微成像系统
CN103616330A (zh) * 2013-11-18 2014-03-05 中国科学院化学研究所 基于超连续产生的宽带激光光源激发的超分辨sted显微成像系统
WO2017174100A1 (de) * 2016-04-08 2017-10-12 Universität Heidelberg Parallelisierung des sted-mikroskopieverfahrens
CN106841149A (zh) * 2017-03-17 2017-06-13 王富 受激辐射损耗显微方法及显微装置
CN108957720A (zh) * 2018-09-26 2018-12-07 中国科学院化学研究所 用于sted光学显微镜的照明系统及sted光学显微镜

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111855544A (zh) * 2020-07-31 2020-10-30 洹仪科技(上海)有限公司 一种荧光成像装置及其成像方法
CN113484320A (zh) * 2021-07-01 2021-10-08 西北大学 一种远场光学超薄片层成像系统及方法
CN115508994A (zh) * 2022-09-14 2022-12-23 郑州思昆生物工程有限公司 一种显微成像系统
CN115728869A (zh) * 2022-11-15 2023-03-03 昂纳信息技术（深圳）有限公司 一种可调谐光滤波器

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