WO2022053649A1 - Microscope optique pour imagerie à résolution spectrale et procédé pour un tel microscope - Google Patents

Microscope optique pour imagerie à résolution spectrale et procédé pour un tel microscope Download PDF

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Publication number
WO2022053649A1
WO2022053649A1 PCT/EP2021/075003 EP2021075003W WO2022053649A1 WO 2022053649 A1 WO2022053649 A1 WO 2022053649A1 EP 2021075003 W EP2021075003 W EP 2021075003W WO 2022053649 A1 WO2022053649 A1 WO 2022053649A1
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WO
WIPO (PCT)
Prior art keywords
image sensor
optical element
emitters
digital image
sample
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Application number
PCT/EP2021/075003
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English (en)
Inventor
Koen Jacobus Adrianus MARTENS
Johannes HOHLBEIN
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Wageningen Universiteit
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Application filed by Wageningen Universiteit filed Critical Wageningen Universiteit
Publication of WO2022053649A1 publication Critical patent/WO2022053649A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361—Optical details, e.g. image relay to the camera or image sensor
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/18—Arrangements with more than one light path, e.g. for comparing two specimens
    • 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
    • G02B27/42—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect
    • G02B27/4233—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect having a diffractive element [DOE] contributing to a non-imaging application
    • G02B27/4238—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect having a diffractive element [DOE] contributing to a non-imaging application in optical recording or readout devices
    • 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
    • G02B27/42—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect
    • G02B27/4233—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect having a diffractive element [DOE] contributing to a non-imaging application
    • G02B27/4244—Diffraction optics, i.e. systems including a diffractive element being designed for providing a diffractive effect having a diffractive element [DOE] contributing to a non-imaging application in wavelength selecting devices

Definitions

  • the present invention relates to an optical microscope comprising a light source, a sample plane, an optical imaging system and a digital image sensor. Additionally, the invention relates to a method for microscopy using such an optical microscope.
  • a known method uses one or more dichroic mirrors that filter the desired spectrum regime and are placed in the emission pathway of the microscope. Next, the emission belonging to lower and higher wavelengths are placed separately on either the same camera chip (side-by-side) or on different cameras. By then observing the ratio in the intensity of high-pass- and low-pass-filtered single-molecule emission pattern, emission characteristics are elucidated.
  • dichroic mirrors are only capable of resolving a limited variety of emitters (wavelength ranges). This leads to limited freedom in the choice of emitters.
  • the emission path is complex by requiring a multi-component set-up in the microscope to create the desired behaviour.
  • a second approach relates to so-called Point Spread Function (PSF) - engineering based on emission.
  • a spatial light modulator (SLM device) can be placed in the emission path of a fluorescence microscope, where it is known that the phase difference is wavelength-dependant. By then designing patterns on the SLM device, differences in emission wavelength can be translated to spatial differences on the camera chip.
  • SLM devices are expensive and are polarisation dependent (leading to 50% loss of all photons), and the emission pathway before and behind the SLM device is complex by requiring a multi-component setup.
  • spectral information is typically spread out over many pixels of the camera chip, leading to poor signal-to-noise (resulting in poor spectral precision) and to low achievable emitter density (due to overlapping spatial/spectral signals).
  • a third approach applies diffraction techniques based on gratings or prisms in an implementation that a ‘spatial channel’ and a ‘spectral channel’ are created (either on the same camera chip or on different cameras). Then, the obtained patterns are fitted with functions to retrieve both the spatial and the spectral information.
  • Bongiovanni 2016 (https://doi.org/10.1038/ncomms13544) uses a transmission grating in front of a detecting camera, but the setup utilises two separated areas on the camera, one for localisation (spatial domain) and one for obtaining spectral information (spectral domain) that are separated into two areas by placing a physical aperture next to the grating leading to a reduction of the effective area on the camera chip for localisation or obtaining spectral information.
  • additional optical components such as mirrors or apertures are required to achieve the separation of the spatial and spectral information on the camera, thereby increasing the minimum accessible distance between grating and camera. All implementations of the latter approach have in common that different areas on the camera are used to obtain spatial and spectral information. Further, the spectral information is spread out over many pixels of the camera limiting the accessible signal to noise.
  • an optical microscope comprising a light source, a sample holder, an optical imaging system and a digital image sensor; the sample holder arranged for holding a spectral emitters-containing sample; the light source configured for emitting a light beam for excitation of associated emitters in the sample; the optical imaging system arranged for collecting radiation emitted by emitters in the sample and imaging an emitters’ image from the collected radiation through the optical imaging system on a light detecting surface of the digital image sensor, wherein the microscope comprises a further optical element for modulating the phase of the radiation emitted from the sample, the further optical element arranged in the detection path before reaching the diffractive optical element, and further comprises a diffractive optical element mounted directly in front of the light detecting surface of the digital image sensor at a first distance; the diffractive optical element is configured for receiving the emitters’ image and creating a diffraction pattern thereof consisting of at least two beams of different order and the first distance is chosen in such a way to allow the at least two beams to be directly projected on
  • the invention relates to a microscope comprising a light source, a sample holder, an optical imaging system and a digital image sensor; the sample holder arranged for holding a spectral emitters-containing sample; the light source configured for emitting a light beam for excitation of associated emitters in the sample; the optical imaging system arranged for collecting radiation emitted by emitters in the sample and imaging an emitters’ image from the collected radiation through the optical imaging system on a light detecting surface of the digital image sensor, wherein the microscope microscope comprises a slit element in the optical path between the sample holder and the optical imaging system and further comprises a diffractive optical element mounted directly in front of the light detecting surface of the digital image sensor at a first distance; the diffractive optical element is configured for receiving the emitters’ image and creating a diffraction pattern thereof consisting of at least two beams of different order and the first distance is chosen in such a way to allow the at least two beams to be directly projected on the light detection surface of the digital image sensor.
  • the diffractive optical element By arranging the diffractive optical element immediately in front of the light detecting surface of the digital image sensor at a distance that allows two or more beams, of which at least one is a diffracted beam, to be directly projected on the light detection surface of the digital image sensor, spatial information and spectral information are both projected in the field of view of the image sensor. No additional optical elements are required between the diffractive optical element and the sensor which simplifies the construction of the microscope.
  • the invention also relates to a method for microscopy, comprising: providing a microscope comprising a light source, a sample holder, an optical imaging system and a digital image sensor; providing a diffractive optical element; arranging the diffractive optical element directly in front of a light detecting surface of the digital image sensor at a first distance thereof; providing an emitters containing sample on/in the sample holder; providing by means of the light source configured for emitting a light beam for providing a selected excitation wavelength for excitation of associated emitters in the sample; by means of the optical imaging system collecting radiation emitted by the emitters and imaging an emitter’s image from the collected radiation along a transmission path on the light detection surface of the digital image sensor, in which a phase of the radiation emitted from the sample is modulated by a further optical element arranged in the detection path before reaching the diffractive optical element, wherein the diffractive optical element produces from the emitters’ image a diffraction pattern comprising at least two diffracted beams from the collected radiation, and the
  • the invention also relates to a method for microscopy, comprising: providing a microscope comprising a light source, a sample holder, an optical imaging system and a digital image sensor; providing a diffractive optical element; arranging the diffractive optical element directly in front of a light detecting surface of the digital image sensor at a first distance thereof; providing an emitters containing sample on/in the sample holder; providing by means of the light source configured for emitting a light beam for providing a selected excitation wavelength for excitation of associated emitters in the sample; by means of the optical imaging system collecting radiation emitted by the emitters and imaging an emitter’s image from the collected radiation along a transmission path on the light detection surface of the digital image sensor, in which a slit element is arranged in the detection path before reaching the diffractive optical element and is configured for letting the collected radiation pass through, wherein the diffractive optical element produces from the emitters’ image a diffraction pattern comprising at least two diffracted beams from the collected radiation, and the at least two
  • Figure 1A, 1 B and 1C schematically show an optical microscope according to a respective embodiment of the invention
  • Figure 2 schematically shows a detail of an optical microscope according to an embodiment of the invention.
  • Figure 3 schematically shows a measurement performed by an optical microscope according to an embodiment of the invention.
  • Figure 1A schematically shows an optical microscope according to an embodiment of the invention.
  • An optical microscope 100 comprises at least a light source 3, a sample holder 5, an optical imaging system 7 and a digital image sensor 9.
  • the light source 3 is configured for emitting a light beam that illuminates the sample 6 that is arranged in or on the sample holder 5.
  • the light source 3 may include optics to project and re-position the light beam 3a on the sample. In this manner, a scanning light beam may be created.
  • the microscope 100 has a widefield/confocal geometry in which the light beam from the light source 3 is directed at the sample plane by means of a polychroic mirror 24 (or dichroic mirror) that is arranged in the optical path between the sample 6 and the optical imaging system 7.
  • An objective lens 4 is arranged between the polychroic mirror 24 and the sample holder 5.
  • the sample holder 5 is arranged to receive the light beam 3a from the light source 3 and to forward light that is either scattered or emitted as fluorescence from emitters in a sample 6 in/on the sample holder 5 towards the image sensor 9.
  • an optical imaging system 7 is positioned between the sample holder 5 and the imaging sensor 9 to collect the forwarded light from the emitters and project an image of a sample on/in the sample holder on the digital image sensor.
  • the optical imaging system 7 comprises one or more optical elements.
  • the optical imaging system 7 consists of a tube lens.
  • the sample holder 5 is a cover glass.
  • the digital image sensor 9 is positioned in a manner that light (i.e., an image of the emitters) after passing the optical imaging system 7 is projected on a light detecting surface 12 thereof.
  • the light detecting surface 12 comprises an array of pixels that each are configured to convert impinging light into an electric signal that can be processed further by a digital processing circuit such as an image processor.
  • the digital image sensor may be a digital sCMOS based camera.
  • the optical microscope comprises a diffractive optical element 14 that is mounted between the optical imaging system 7 and the light detecting surface 12 of the digital image sensor.
  • the diffractive optical element 14 is configured to create a diffraction pattern from the forwarded light (the image of the emitters).
  • the diffractive optical element is positioned close to the light detecting surface in a setup distance 16 that from at least the image of one emitter the diffracted image comprising at least two beams 20, 22 is directly projected on the light detecting surface 12 of the digital image sensor 9.
  • the diffractive optical element 14 may be a diffraction grating, optionally the diffraction grating is a blazed grating.
  • the optical microscope comprises a light source, a sample holder, an optical imaging system and a digital image sensor; the sample holder arranged for holding a spectral emitters-containing sample; the light source configured for emitting a light beam for excitation of associated emitters in the sample; the optical imaging system arranged for collecting radiation emitted by emitters in the sample and imaging an emitters’ image from the collected radiation through the optical imaging system on a light detecting surface of the digital image sensor, wherein the microscope further comprises a diffractive optical element mounted directly in front of the light detecting surface of the digital image sensor at a first distance; the diffractive optical element is configured for receiving the emitters’ image and creating a diffraction pattern thereof consisting of at least two beams of different order and the first distance is chosen in such a way to allow the at least two beams to be directly projected on the light detection surface of the digital image sensor.
  • the microscope comprises an optical element 26 for modulating the phase of the radiation emitted from the sample.
  • Such optical element 26 can be arranged at any position in the optical detection path before reaching the diffractive optical element.
  • the optical element 26 is arranged in the optical path between the polychroic mirror 24 and the diffractive optical element 14.
  • the optical element 26 can be arranged in the optical path between the polychroic mirror 24 and the optical imaging system 7, or between the optical imaging system 7 and the diffractive optical element 14.
  • the optical element 26 for modulating the phase comprises a cylindrical lens.
  • the invention relates to a microscope comprising a light source, a sample holder, an optical imaging system and a digital image sensor; the sample holder arranged for holding a spectral emitters-containing sample; the light source configured for emitting a light beam for excitation of associated emitters in the sample; the optical imaging system arranged for collecting radiation emitted by emitters in the sample and imaging an emitters’ image from the collected radiation through the optical imaging system on a light detecting surface of the digital image sensor, wherein the microscope comprises a further optical element for modulating the phase of the radiation emitted from the sample, the further optical element arranged in the detection path before reaching the diffractive optical element, and the microscope comprises a diffractive optical element mounted directly in front of the light detecting surface of the digital image sensor at a first distance; the diffractive optical element is configured for receiving the emitters’ image and creating a diffraction pattern thereof consisting of at least two beams of different order and the first distance is chosen in such a way to allow the at least two beams to
  • the invention allows to project two or more beams, of which at least one is a diffracted beam, directly on the light detection surface of the digital image sensor, spatial information and spectral information are both projected in the field of view of the image sensor.
  • the optical element 26 provides that the phase of the radiation emitted from the sample towards the detector can be modulated so as to enhance the optical signal received at the digital image sensor.
  • an optical path 18 between the diffractive optical element 14 and the digital image sensor 9 for the direct projection of the at least two beams 20, 22 no additional optical elements are provided.
  • the optical path 18 between the diffractive optical element and the digital image sensor is free from elements causing any interaction with the at least two beams 20, 22.
  • the diffractive optical element is positioned directly in front of the digital image sensor.
  • the two beams 20, 22 may be a non-diffracted beam 20 (a beam passing the diffractive optical element in a direction without diffraction, also referred to as zeroth order diffracted beam) and a diffracted beam 22 diffracted in the direction of the first order maximum of the diffraction pattern, also referred to as first order diffracted beam.
  • the two beams may be two first order diffracted beams on opposite sides of the direction of the zeroth order.
  • the two beams may also comprise other higher order diffracted beams (i.e., higher than first order) that may be projected on the image sensor 9 by means of the diffractive optical element 14.
  • the diffraction pattern is determined by a periodic pattern of the diffractive optical element 14 and the wavelength of the light forwarded from the sample holder 5, i.e., the sample plane, the diffraction pattern will be different for various wavelengths.
  • the distance 16 of the diffractive optical element 14 is optionally configured to be adjustable relative to the light detecting surface, to adapt a distance between the at least two beams that are projected on the light detecting surface.
  • the distance between the diffractive optical element and the light detecting surface can be set minimal to let the diffractive optical element abut (i.e., is mounted directly on) the light detection surface of the digital image sensor.
  • the light source 3 comprises at least one laser beam source to create a monochrome light beam (for example 561 nm or 642 nm).
  • the laser beam source may be a laser beam source with tuneable wavelength.
  • the light source 3 may comprise multiple laser beam sources, for example two or three or more, each with a different wavelength to create a combined light beam comprising multiple wavelength components.
  • the light source 3 may comprise a light source capable of creating a light beam with a small spectral bandwidth, for example light within a bandwidth of 50 nm or less, around a central wavelength.
  • the microscope 100 is a fluorescence microscope or a single-molecule fluorescence capable microscope for analysing samples that comprise one or more emitter substances as additive in the sample that fluoresce or scatter at respective different wavelengths due to excitation by light (or light component) from the light source 3.
  • the forwarded light from the sample thus may comprise one or more wavelength components depending on the presence of different types of emitters.
  • Emitter substances can be selected for example from fluorophores or nanoparticles used for inelastic scattering or plasmonics.
  • the microscope is provided with a slit element 10 for shaping the forwarded light beam.
  • the slit element 10 (drawn as a dashed line) is typically arranged at any position in the optical path between the polychroic mirror 24 and the diffractive optical element 14.
  • the slit element 10 is arranged in the optical path between the sample holder 5 and the optical imaging system 7. In a further embodiment, the slit element 10 is arranged in the optical path between the polychroic mirror 24 and the optical imaging system 7, as shown in fig 1A.
  • the slit element 10 is provided to the microscope in addition to the optical element 26 for phase modulation of the radiation.
  • the slit element 10 is provided to the microscope as an alternative for the optical element 26 for phase modulation of the radiation.
  • the invention relates to a microscope which comprises a light source, a sample holder, an optical imaging system and a digital image sensor; the sample holder arranged for holding a spectral emitters-containing sample; the light source configured for emitting a light beam for excitation of associated emitters in the sample; the optical imaging system arranged for collecting radiation emitted by emitters in the sample and imaging an emitters’ image from the collected radiation through the optical imaging system on a light detecting surface of the digital image sensor, wherein the microscope microscope comprises a slit element in the optical path between the sample holder and the optical imaging system and further comprises a diffractive optical element mounted directly in front of the light detecting surface of the digital image sensor at a first distance; the diffractive optical element is configured for receiving the emitters’ image and creating a diffraction pattern thereof consisting of at least two beams of different order and the first distance is chosen in
  • the slit element 10 provides a shaping of the radiation emitted from the sample towards the detector so as to enhance the optical signal received at the digital image sensor.
  • FIG 1B schematically shows an optical microscope 101 according to an embodiment of the invention.
  • the optical microscope 101 is very similar to the optical microscope 100 as shown in Figure 1A and comprises as above described the light source 3, the sample holder 5, the optical imaging system 7 and the digital image sensor 9.
  • the light source 3 is positioned above the sample holder 5 to directly illuminate the emitters in the sample 6.
  • the optical imaging system 7 is positioned between the sample holder 5 and the imaging sensor 9 to collect the forwarded light from emitters in the sample 6 and project an image of the emitters on the digital image sensor.
  • the optical microscope comprises a diffractive optical element 14 that is mounted between the optical imaging system 7 and the light detecting surface 12 of the digital image sensor, close to the light detecting surface 12.
  • the microscope 101 further comprises a further optical element for modulating the phase of the radiation emitted from the sample, the further optical element arranged in the detection path before reaching the diffractive optical element.
  • the microscope comprises a slit element 10 in the optical path between the sample holder 5 and the optical imaging system 7 or in the optical path between the polychroic mirror 24 and the optical imaging system 7.
  • the diffractive optical element 14 and the digital image sensor 9 for the direct projection of the at least two beams 20, 22 no additional optical elements are provided.
  • an optical element for modulating the phase of the radiation emitted from the sample and/or a slit element 10 can be arranged in the optical path between the sample plane 5 and the diffractive optical element 14 or in a further embodiment in the optical path between the polychroic mirror 24 and the optical imaging system 7.
  • FIG. 1C schematically shows an optical microscope 102 according to an embodiment of the invention.
  • the optical microscope 102 is very similar to the optical microscopes 100; 101 as described above, and accordingly comprises the light source 3, the sample holder 5, the optical imaging system 7 and the digital image sensor 9, the further optical element 26 and/or the slit element 10.
  • the sample plane is provided with a waveguide 8 for illuminating emitters in a sample 6 on the sample holder 5.
  • the output of the light source 3 is coupled to an input of the waveguide 8 for illumination of the sample 6.
  • the sample holder 5 and the waveguide 8 are integrated.
  • Figure 2 schematically shows a detail of the microscopes of Figure 1A-1C.
  • the position of the diffractive optical element 14 is adjustable relative to the light detecting surface 12, as indicated by the double arrow 28.
  • the digital image sensor 9 is a digital sCMOS based camera and the diffractive optical element 14 is installed in a threaded retainer ring mounted on the camera in position direct in front of the light detecting surface 12 of the digital image sensor 9.
  • a distance 16 between the diffractive optical element 14 and the light detecting surface 12 can be adapted.
  • the diffractive optical element 14 is positioned in front of the light detecting surface 12 at a distance 16, without providing any additional optical element therebetween.
  • a zeroth order diffracted beam 20 i.e. , an non-diffracted beam passing the diffractive optical element 14 in non-diffracted direction of a zeroth order maximum
  • a first order diffracted beam 22 i.e., radiation running in the direction of the first maximum in the diffraction pattern next to the zero order beam
  • the projections of the diffracted beams are spaced apart over a lateral distance 30 depending on the diffraction angle between the zeroth and first (or other orders) (in general, depending on the difference of the diffraction angles of the observed diffraction orders).
  • the lateral distance 30 will change accordingly. This allows to adapt the size of the diffraction pattern relative to the light detecting surface area of the image sensor 9 for example to enhance the resolution of the projected diffracted beams.
  • the microscope 100; 101 ; 102 may comprise, or may be coupled to, an image processing system 32 to analyse images captured by the digital image sensor 9, for example to determine the lateral distance between any pair of diffracted beams, or to determine the intensity distribution in the projections of the diffracted beams.
  • pairs of diffracted beams can be projected on the light detecting surface: for example the minus first order diffracted beam and the positive first order diffracted beam (i.e. , the first diffraction maxima on both sides of the zero order maximum) may be projected and analysed with respect to their lateral distance and intensity distribution.
  • images of multiple emitters can be projected simultaneously.
  • the grating When using a diffractive grating as diffractive optical element, the grating may have a density between about 20 and about 2000 lines/mm.
  • Figure 3 schematically shows a measurement of an emitter substance containing sample performed by a microscope according to an embodiment of the invention.
  • Figure 3 a top view of the light detecting surface 12 of the image sensor 9 is shown while a sample is illuminated by light from the light source 3 of the microscope.
  • the sample emitters are present at certain locations, which are configured for emitting radiation due to an excitation by light received from the light source. Since the light from the light source is selectively filtered out by a suitable regular emission filter and the radiation from the emitters can pass the regular emission filter, an image of the locations of the radiating emitters in the sample is selectively transmitted through the optical imaging system towards the image sensor. After passing the diffractive optical element, the images 34; 37 of individual emitters are projected without diffraction (as zero order diffracted intensity) on the light detecting surface presenting spatial information.
  • the diffracted image 35; 38 of each emitter is projected as first order diffraction beam (e.g., +1 order) in the direction of the first order diffraction maximum offset over a lateral distance 36; 39 with respect to the zero order 34; 37.
  • the lateral distance is dependent on the emission wavelength.
  • any spectral variation in the radiation from the emitter will be visible by a broadening of the projection of the first order diffracted beams in the diffraction direction on the light detecting surface.
  • the spectral characteristics of the projection of the diffracted beam can be derived from the intensity distribution detected by individual pixels in the sensor. In case of multiple emitters being imaged the spatial information of these emitters is projected all over the image sensor, as well as the diffracted beams corresponding with these emitters. For each emitter, the diffracted beam is simply offset in the diffraction direction relative to the location of the image of the respective emitter by a spectrumdependant distance, but both spatial and spectral information are projected on the whole of the sensor.
  • the distance between the zeroth (or other) order and the first (or other) order diffraction beam can be determined from the image captured by the image sensor. It will be appreciated that also unpaired zeroth order maxima 40 can be detected on the digital image sensor if the associated first order maximum 41 is not projected on the digital image sensor. Likewise, an unpaired first order maximum 42 can be detected by the digital image sensor, without the associated zeroth order maximum 43 being projected on the digital image sensor.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention concerne un microscope comprenant une source de lumière, un porte-échantillon, un système d'imagerie optique et un capteur d'image numérique. Le porte-échantillon est conçu pour contenir un échantillon contenant des émetteurs spectraux avec des profils d'émission spécifiques. La source de lumière est configurée pour émettre un faisceau de lumière pour l'excitation d'émetteurs dans l'échantillon. Le système d'imagerie optique est conçu pour collecter un rayonnement provenant d'émetteurs dans l'échantillon et imager une image d'émetteurs à partir du rayonnement collecté à travers le système d'imagerie optique sur le capteur d'image numérique. Le microscope comprend un élément optique diffractif monté directement devant le capteur d'image numérique à une première distance. L'élément optique diffractif est configuré pour recevoir l'image d'émetteurs et créer un motif de diffraction de celle-ci d'au moins deux faisceaux d'ordre différent. La première distance est choisie de telle sorte que les au moins deux faisceaux sont directement projetés sur le capteur d'image numérique.
PCT/EP2021/075003 2020-09-10 2021-09-10 Microscope optique pour imagerie à résolution spectrale et procédé pour un tel microscope WO2022053649A1 (fr)

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EP20195448.4 2020-09-10

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060012785A1 (en) * 2004-07-16 2006-01-19 Joerg-Michael Funk Light scanning electron microscope and use
US20170176338A1 (en) * 2015-12-21 2017-06-22 Verily Life Sciences Llc Spectrally And Spatially Multiplexed Fluorescent Probes For In Situ Cell Labeling
US20170307440A1 (en) * 2014-09-25 2017-10-26 Northwestern University Devices, methods, and systems relating to super resolution imaging

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060012785A1 (en) * 2004-07-16 2006-01-19 Joerg-Michael Funk Light scanning electron microscope and use
US20170307440A1 (en) * 2014-09-25 2017-10-26 Northwestern University Devices, methods, and systems relating to super resolution imaging
US20170176338A1 (en) * 2015-12-21 2017-06-22 Verily Life Sciences Llc Spectrally And Spatially Multiplexed Fluorescent Probes For In Situ Cell Labeling

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BONGIOVANNI MARIE N. ET AL: "Multi-dimensional super-resolution imaging enables surface hydrophobicity mapping", vol. 7, no. 1, 1 December 2016 (2016-12-01), XP055774589, Retrieved from the Internet <URL:http://www.nature.com/articles/ncomms13544> [retrieved on 20210210], DOI: 10.1038/ncomms13544 *
CHRISTENSEN D ET AL: "ANALYSIS OF EXCITATION AND COLLECTION GEOMETRIES FOR PLANAR WAVEGUIDE IMMUNOSENSORS", PROCEEDINGS OF SPIE, IEEE, US, vol. 1886, 21 January 1993 (1993-01-21), pages 2 - 08, XP000672419, ISBN: 978-1-62841-730-2, DOI: 10.1117/12.144833 *

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