EP3953684A1 - Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons - Google Patents

Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons

Info

Publication number
EP3953684A1
EP3953684A1 EP20719335.0A EP20719335A EP3953684A1 EP 3953684 A1 EP3953684 A1 EP 3953684A1 EP 20719335 A EP20719335 A EP 20719335A EP 3953684 A1 EP3953684 A1 EP 3953684A1
Authority
EP
European Patent Office
Prior art keywords
light sheet
light
cover
designed
support glass
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP20719335.0A
Other languages
German (de)
English (en)
Inventor
Alexander Weiss
Christian Schumann
Ronja CAPELLMANN
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems CMS GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Leica Microsystems CMS GmbH filed Critical Leica Microsystems CMS GmbH
Publication of EP3953684A1 publication Critical patent/EP3953684A1/fr
Pending legal-status Critical Current

Links

Classifications

    • 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
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/33—Immersion oils, or microscope systems or objectives for use with immersion fluids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • 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
    • G02B21/0004—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
    • G02B21/008—Details of detection or image processing, including general computer control
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/02—Objectives
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/082—Condensers for incident illumination only
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/10—Condensers affording dark-field illumination
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/241—Devices for focusing
    • G02B21/245—Devices for focusing using auxiliary sources, detectors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides

Definitions

  • the invention relates to a light sheet microscope.
  • the invention also relates to a method for acquiring a measured variable by means of a light sheet microscope.
  • the thickness of the cover glass since this determines the length of the optical path which the detection light to be detected by the objective traverses when passing through the cover glass.
  • Another optical parameter is the distance between the cover or support glass and the imaging lens. Knowledge of this distance is necessary, for example, to implement an autofocus system.
  • the document US Pat. No. 8,5082,203 B2 discloses a light sheet microscope with an illumination device for generating a light sheet and an objective.
  • the light sheet is arranged in a sample in such a way that the direction of propagation of the illuminating light that forms the light sheet is not perpendicular to the optical axis of the objective of the microscope.
  • Such light sheet microscopes are therefore also referred to as oblique plane microscopes (OPM: oblique plane microscope).
  • the document DE 10 2016 119 268 B3 discloses a light sheet microscope with a lighting device that generates a light sheet in an intermediate image space, and a transport optics that images the light sheet into the sample.
  • the documents WO 2017/210159 A1 and WO 2015/109323 A2 each disclose a light sheet microscope with an illumination device which is designed to generate a light sheet and move it laterally through a sample. Such light sheet microscopes are also referred to as SCAPE microscopes (SCAPE: swept confocally-aligned planar excitation).
  • the light sheet microscope comprises a sample space in which a cover or support glass can be arranged which has a surface that defines a partially reflective interface, an optical system with an objective facing the cover or support glass, an illumination device that is designed, a light sheet to generate a sensor, and a processor.
  • the two boundary surfaces are formed by the fact that two optical media can be attached to the sample space, which media adjoin the two surfaces of the cover or support glass.
  • the light sheet microscope is designed in such a way that it forms a measuring device for detecting a measured variable.
  • the measuring device is designed to direct the light sheet through the optics system at an oblique incidence onto the cover or support glass, to generate a reflected light bundle by partially reflecting the light sheet at the interface, and to receive and receive the reflected light bundle through the optics system to steer towards the sensor.
  • the sensor is designed to detect the intensity and / or the point of incidence of the reflected light bundle.
  • the processor is designed to determine the measured variable on the basis of the detected intensity and / or the location of incidence of the reflected light bundle.
  • cover or support glass is understood to mean, in particular, a cover glass covering a sample, a slide, the bottom of a Petri dish or the bottom of a cavity of a microtiter plate.
  • the surface of the cover or support glass which forms the interface can be either a surface facing the objective or a surface of the cover or support glass facing away from the objective.
  • the sensor is preferably designed as a position-sensitive sensor, i.e. as a sensor that is suitable for detecting the intensity and the point of incidence.
  • the light sheet microscope uses the reflected light beam, i. a partial reflection of the light sheet on the surface of the cover or support glass in order to record the measured variable.
  • the reflected light bundle used to determine the measured variable therefore has a wavelength that generally differs by a few nanometers, the so-called Stokes shift, from the wavelength of the detection light emanating from the sample.
  • the Stokes shift is about 5 to 20 nm, and even more in the case of unusual fluorophores. This means in particular that when a wavelength-dependent measured variable is determined, no conversion of the measured variable to the wavelength of the detection light is required and no assumption about the dispersion needs to be made. This enables a particularly reliable determination of the measured variable.
  • those measured variables are wavelength-dependent that are required to determine an imaging error, for example the length of an optical path through the cover or support glass or a refractive index.
  • the proposed light sheet microscope can thus be used to improve the imaging quality of the light sheet microscope by setting suitable manipulated variables of the light sheet microscope on the basis of the measured value determined.
  • the partial reflection of the light sheet also occurs in previously known light sheet microscopes. It represents a disturbance that is separated from the detection light by means of a blocking filter.
  • the proposed light sheet microscope thus makes particularly advantageous use of a disturbance variable that is disadvantageous per se to improve the image quality.
  • the solution described here can therefore be implemented with little effort by appropriately configuring an existing light sheet microscope.
  • the light sheet thus fulfills the same function as a measuring light beam formed by means of a slit diaphragm in known autofocus devices, for example according to DE 10 2010 0S0 430
  • the processor is designed to determine the distance of the cover or support glass from the objective along its optical axis as a measured variable on the basis of the detected point of incidence of the reflection light bundle.
  • the measuring device can be used as an autofocus device or as part of such.
  • Each of the at least three measuring points is determined by three coordinates, one of which indicates the distance to be determined between the measuring point and the lens along its optical axis, while the other two coordinates define the position of the respective measuring point on the surface of the cover glass .
  • the cover or support glass has a further surface which defines a further partially reflective interface.
  • the two boundary surfaces are arranged at different distances from the objective.
  • the measuring device in this embodiment is designed to generate a further bundle of reflected light by partially reflecting the light sheet at the further boundary surface and to receive the further bundle of reflected light through the objective and direct it onto the sensor.
  • the sensor is designed to detect the intensity and / or the location of incidence of the further reflected light bundle.
  • the processor is designed to determine the measured variable on the basis of the recorded intensities and / or incidence locations of the two reflected light bundles.
  • the measuring device is designed to direct the two reflected light bundles simultaneously onto the detector and to determine the thickness of the cover or support glass based on the mutual spacing of the different incidence locations of the two reflected light bundles.
  • This special embodiment is designed in particular for cover or support glasses which are relatively thin in relation to the lens magnification. In this case there is the spatial separation of the two The reflected light bundle is correspondingly small, so that both can be detected on the detector at the same time.
  • the measuring device is designed to direct the two reflected light bundles one after the other onto the detector. This means that the measuring device detects only one of the reflected light bundles on the detector at a given point in time.
  • Such an embodiment is designed in particular for cover or support glasses which are comparatively thick with respect to the objective magnification. In this case, the mutual distance between the two boundary surfaces at which the reflection light bundles arise is so great that the associated spatial separation of the reflection light bundles makes it impossible to receive both reflection light bundles at the same time on the detector.
  • the processor is preferably designed to set the distance between the cover or support glass and the objective as an operating parameter.
  • This distance, measured along the optical axis of the objective can be changed, for example, using a suitable adjustment device, such as a so-called z-drive.
  • a suitable adjustment device such as a so-called z-drive.
  • the measuring device is designed to set the position of a displaceable focusing lens provided in the light sheet microscope as an operating parameter. From the change in position of this displaceable focusing lens, the optical thickness of the cover or support glass can then be determined as the measured variable, taking into account the optical imaging conditions.
  • the processor is designed to use the optical thickness to determine a mechanical thickness of the cover or support glass, taking into account the refractive indices of the cover or support glass and an optical medium that is attached to both the objective and the cover. or glass is adjacent to be determined as a measured variable.
  • the aforementioned optical medium is, for example, an immersion medium that is located between the cover or support glass and the objective.
  • the numerical aperture of the main beam of the light sheet in the sample space is also taken into account. This is preferably done according to the following relationship: where NA denotes the numerical aperture of the main ray of the light sheet in the sample space.
  • the numerical aperture NA is given by the product of the refractive index ni m of the immersion medium and the angle of incidence at which the measuring light beam emerging from the objective falls on the cover or support glass with respect to the optical axis.
  • the usual imaging errors can be taken into account when calculating the mechanical thickness.
  • the two boundary surfaces are formed in that two optical media in the sample space adjoin the two surfaces of the cover or support glass.
  • the processor is designed to determine the refractive index of one of the two optical media as a measured variable on the basis of the recorded intensities of the two reflected light bundles.
  • the intensities of the two reflected light bundles depend on the reflection and the transmission of the light sheet at the two interfaces, which are defined by the cover or support glass and the two optical media that border the cover or support glass from opposite sides.
  • the reflection and transmission processes, on which the intensities of the two spatially separated reflection light bundles are ultimately based, are thus essentially determined by the refractive indices of the cover or support glass and the optical media adjoining the cover or support glass . If the refractive index of the cover or support glass and the refractive index of one of the two adjacent optical media are known, the refractive index of the other medium can be reliably determined from the intensities recorded by the detector and the knowledge of the angle of incidence of the measuring light beam in the sample space .
  • the spatial separation of the two reflected light bundles results from the fact that the measuring device directs the light sheet obliquely onto the cover or support glass.
  • the two partially reflective interfaces are axially offset from one another, ie are spaced apart along the optical axis of the lens, the oblique incidence of the light sheet on the two interfaces ensures that the two reflective light bundles are reflected back into the lens on different optical paths.
  • the two reflected light bundles on the detector are recorded separately from one another at different points of incidence.
  • the processor is preferably designed to determine the refractive index of the one optical medium on the basis of the ratio of the intensities of the two reflected light bundles as a measured variable.
  • the measurement of the refractive index is, as it were, self-referential. This means that the refractive index can be determined independently of the intensity of the measuring light beam, i.e. no knowledge of this intensity is required.
  • the optical medium, the refractive index of which is to be determined as a measured variable by means of the measuring device is preferably an embedding medium for a sample which borders on one of the two surfaces of the cover or support glass.
  • the other optical medium, the refractive index of which is known from the outset is preferably an immersion medium that is adjacent to the other surface of the cover or support glass on the one hand and the objective on the other.
  • the measuring device it is also possible to use the measuring device to determine the refractive index of any medium, provided that it is directly adjacent to one of the two surfaces of the cover or support glass and thereby forms a partially reflective interface.
  • the aforementioned intensity distribution is obtained by integrating the respective image on the detector over a direction that is parallel to the longitudinal orientation of the cross-section of the Light sheet lies.
  • the light sheet microscope includes a detection device with a further sensor for detecting the detection light emanating from the sample and a beam splitter unit which is designed to direct the reflected light bundle onto the sensor and the detection light onto the further sensor.
  • the light sheet microscope comprises two sensors, each of which has only one function. This allows the use of specialized sensors and thus a more reliable detection of both the reflected light bundle and the detection light. This makes the determination of the measured variable more reliable.
  • the light sheet microscope comprises a filter which can be connected upstream of the sensor and which is impermeable to the reflected light bundle. This prevents the reflected light bundle from influencing the detection of the detection light emanating from the sample.
  • the switchability of the filter means that the sensor can be used both to record the reflected light beam and the detection light.
  • the light sheet microscope preferably comprises a scanning element which is designed to move the light sheet along a scanning axis.
  • the scanning element is, for example, a movable raster mirror.
  • the light sheet microscope has an electronic focusing device which is designed to focus the light sheet on different planes in the sample space. This also allows the light sheet to be moved along the scanning axis.
  • the light sheet microscope is often also referred to as OPM (OPM: oblique plane microscope). If the scanning takes place laterally to the optical axis of the objective, the light sheet microscope is often referred to as a SCAPE microscope (SCAPE: swept confocally-aligned planar excitation).
  • SCAPE swept confocally-aligned planar excitation
  • the invention also relates to a method for acquiring a measured variable by means of a light sheet microscope.
  • the method has the advantages described above and can be developed in the same way, in particular by the features of the dependent claims related to the light sheet microscope.
  • FIG. 1 shows a schematic representation of a light sheet microscope which provides scanning lateral to the optical axis of the objective, according to an embodiment
  • FIG. 2 shows a schematic representation of the light sheet microscope, which provides a scanning lateral to the optical axis of the objective, according to an embodiment with two sensors;
  • FIG. 3 shows a schematic representation of the light sheet microscope, which provides a scanning along the optical axis of the objective, according to an embodiment
  • FIG. 4 shows a schematic representation of the light sheet microscope, which provides scanning along the optical axis of the objective, according to an embodiment with two sensors;
  • FIG. 5 shows a schematic illustration showing a sample space of the light sheet microscope
  • FIG. 6 shows an intensity distribution detected by a position-sensitive detector of the light sheet microscope with an intensity maximum
  • FIG. 7 shows a further schematic illustration which shows the sample space of the light sheet microscope.
  • FIG. 1 shows a schematic illustration of a light sheet microscope 100 in a special embodiment which provides scanning lateral to the optical axis 02 of an objective 120.
  • the light sheet microscope 100 enables, in addition to imaging using light sheet microscopy, the acquisition of a measured variable that influences the imaging.
  • the light sheet microscope 100 at the same time forms a measuring device for this measured variable.
  • the light sheet microscope 100 comprises an illumination device 102, a transport optics 104 and a detection device 106, which together form an optics system 107.
  • the optical axis OB of the detection device 106 is tilted by a tilt angle ⁇ with respect to the optical axis 02 of the transport optics 104.
  • the optical Axis 01 of the lighting device 102 is perpendicular to the optical axis OB of the detection device 106, so that the optical axis 01 of the lighting device 102 is tilted by an angle of 90 ° with respect to the optical axis 02 of the transport optics 104.
  • the three aforementioned optical axes 01, 02, 03 intersect in an intermediate image space 108.
  • a cover or support glass which is simply referred to as cover glass 118 in the following, and two optical media 117, 119 arranged, each of which adjoins the cover glass 118.
  • the light sheet microscope 100 also has a processor 110.
  • the lighting device 102 comprises a light source 112 and an illuminating objective 114.
  • the light source 112 generates a light sheet, for example with the aid of a cylinder lens not explicitly shown in FIG. 1, the light sheet with reference to an oblique coordinate system shown in FIG a direction of light propagation A and a direction of expansion B perpendicular thereto.
  • the light source 112 can also generate a quasi-static light sheet by means of a scanning element specially provided for this purpose.
  • the lighting objective 114 images the light sheet in the intermediate image space 108.
  • the light sheet can also be coupled directly into the transport optics 104.
  • the transport optics 104 viewed from the sample space 116, comprise the objective 120 facing the cover glass 118, a first tube lens 122, a first eyepiece 124, a second eyepiece 126, a second tube lens 128 and a projection objective 130, which are positioned along the optical axis 02 the transport optics 104 are arranged.
  • a scanning device is arranged between the first eyepiece 124 and the second eyepiece 126, which in the embodiment shown is formed by a raster mirror 132 on which the optical axis 02 of the optical transport system 104 is deflected.
  • the processor 110 is connected to the light source 112, the raster mirror 132, the position-sensitive sensor 150 and a so-called z-drive 111 which, for example by moving a microscope table, determines the position of the cover glass 118 along the optical axis 02 of the Lens 120 can change.
  • the processor 110 is designed to determine the measured variable on the basis of the detected intensity and / or the location of incidence of the reflection light bundle 142.
  • the processor 110 also has a memory 154 in which parameters for determining the measured variable can be stored.
  • FIG. 2 shows a schematic representation of a light sheet microscope 200 which provides scanning lateral to the optical axis 02 of the objective 120.
  • the light sheet microscope 200 represents a modified embodiment with two sensors 150, 202.
  • the light sheet microscope 200 according to FIG. 2 thus differs from the embodiment according to FIG. 1 essentially in the further sensor 202 and a beam splitter unit 204, which is designed to direct the reflection light bundle 142 onto the position-sensitive sensor 150 and detection light onto the further sensor 202.
  • Identical and identically acting elements are denoted by the same reference symbols in FIGS. 1 and 2.
  • FIG. 3 shows a light sheet microscope 300 as a modified embodiment, which differs from the embodiment according to FIG. 1 essentially in that the scanning through the light sheet is axial, i.e. takes place along the optical axis 02 of the transport optics 104.
  • Identical and identically acting elements are denoted by the same reference symbols in FIGS. 1 to 3.
  • FIG. 4 shows a schematic representation of a light sheet microscope 400 that provides scanning along the optical axis 02 of the objective 120, according to an embodiment with two sensors 150, 202.
  • the embodiment of the light sheet microscope 400 according to FIG. 4 differs from the embodiment of the light sheet microscope 300 according to FIG. 3 essentially by the further sensor 202 and the beam splitter unit 204.
  • Identical and identically acting elements are denoted by the same reference numerals in FIGS.
  • FIG. 5 is a schematic illustration which shows the sample space 116 of the light sheet microscope 100, 200, 300, 400.
  • FIG. 5 shows how the reflected light bundle 142 is generated by reflecting the light sheet 134 at first one of the surfaces 138, 140 of the cover glass 118.
  • the light sheet 134 which decentrally illuminates the entrance pupil 136 of the objective 120, is directed by the objective 120 at an angle ⁇ obliquely to the optical axis 02 onto the front surface of the cover glass 118 facing the objective 120 and denoted by 138 in FIG . Since the cover glass 118 and an immersion medium 119 adjoining its front surface 138 have different refractive indices, the front surface 138 of the cover glass 118 and the immersion medium 119 adjoining it form an interface at which the incident light sheet 134 is partly reflected. The part of the measuring light bundle 134 that is reflected at this interface generates the reflection light bundle 142, which is guided back into the objective 120.
  • the other part 152 of the light sheet 134 which transmits the first interface, is broken away from the optical axis 02 of the objective 120 when it enters the cover glass 118 and forms an angle g with this which is greater than the angle ⁇ .
  • This transmitted part 152 of the light sheet 134 is partly reflected at a second boundary surface which is defined by the rear surface 140 of the cover glass 118 and an adjoining embedding medium 117 which has a different refractive index than the cover glass 118.
  • This second partial reflection of the light sheet 134 at the second boundary surface generates the second reflection light bundle 142b, which passes through the front surface 138 of the cover glass 118 and then passes back into the objective 120.
  • the oblique incidence of the light sheet 134 into the sample space 116 ensures that the reflection light bundles 142a, 142b generated by the two partial reflections on the front surface 138 and the rear surface 140 of the cover glass 118 point to different optical paths back into the objective 120.
  • the two reflection light bundles 142a, 142b strike the position-sensitive detector 150 at different points of incidence, provided that it is ensured that both reflection light bundles 142a, 142b fall simultaneously on the detector 150.
  • the two images of the cross section of the light sheet 134 generated on the front surface 138 and the rear surface 140 of the cover glass 118 are mapped onto the position-sensitive detector 150 spatially separated from one another, as illustrated in the following diagram according to FIG.
  • the second partial reflection takes place on the rear surface 140 of the cover glass 188 at a point offset transversely to the optical axis O2.
  • the areas below the peaks PI, P2 shown in FIG. 8 are each a measure of the intensity of the respective reflection light bundle 142a, 142b. From their ratio, for example, the refractive index of one of the two optical media 117, 119 can be determined as a measured variable.
  • FIG. 8 shows a situation in which the two reflection light bundles 142a, 142b fall simultaneously on the position-sensitive detector 150. This means that the spatial splitting of the two reflection light bundles 142a,
  • the thickness of the cover glass 118 to be detected as a measured variable is relatively small in relation to the objective magnification.
  • the spatial splitting of the reflection light bundles 142a, 142b, corresponding to the thickness of the cover glass 118 to be detected is so great that a simultaneous detection of the two reflection light bundles 142a, 142b by the position-sensitive detector 150 is not possible is.
  • aspects have been described in the context of a device, it is clear that these aspects also represent a description of the corresponding method, with a block or a device corresponding to a method step or a function of a method step. Analogously, aspects that are described in the context of a method step also represent a description of a corresponding block or element or a property of a corresponding device.
  • Some or all of the method steps can be carried out by (or using) a hardware device, as described in FIG Example can be a processor, a microprocessor, a programmable computer or an electronic circuit.
  • one or more of the most important method steps can be carried out by such a device.
  • embodiments of the invention can be implemented in hardware or software.
  • the implementation can be carried out with a non-volatile storage medium such as a digital storage medium such as a floppy disk, a DVD, a Blu-Ray, a CD, a ROM, a PROM and EPROM, an EEPROM or a FLASH memory, are carried out on the electronically readable control signals are stored, which interact (or can cooperate) with a programmable computer system so that the respective method is carried out. Therefore, the digital storage medium can be computer readable.
  • Some exemplary embodiments according to the invention comprise a data carrier with electronically readable control signals which can interact with a programmable computer system so that one of the methods described herein is carried out.
  • exemplary embodiments of the present invention can be implemented as a computer program product with a program code, the program code being effective for executing one of the methods when the computer program product is running on a computer.
  • the program code can, for example, be stored on a machine-readable carrier.
  • Another exemplary embodiment of the invention is therefore a data stream or a signal sequence which represents the computer program for carrying out one of the methods described herein.
  • the data stream or the signal sequence can for example be configured in such a way that they are transmitted via a data communication connection, for example via the Internet.
  • Another embodiment comprises a processing means, for example a computer or a programmable logic device, which is configured or adapted to carry out one of the methods described herein.
  • a processing means for example a computer or a programmable logic device, which is configured or adapted to carry out one of the methods described herein.
  • Another exemplary embodiment comprises a computer on which the computer program for carrying out one of the methods described herein is installed.
  • a further exemplary embodiment according to the invention comprises a device or a system which is configured to transmit (for example electronically or optically) a computer program for carrying out one of the methods described herein to a receiver.
  • the receiver can for example be a computer, a mobile device, a storage device or the like.
  • the device or the system can for example comprise a file server for transmitting the computer program to the recipient.
  • a programmable logic device eg, a field programmable gate array, FPGA
  • FPGA field programmable gate array
  • a field programmable gate arrangement can cooperate with a microprocessor to perform any of the methods described herein. In general, the methods are preferably performed by any hardware device.

Landscapes

  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Engineering & Computer Science (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Multimedia (AREA)
  • Pathology (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • General Engineering & Computer Science (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Length Measuring Devices By Optical Means (AREA)
  • Focusing (AREA)
  • Automatic Focus Adjustment (AREA)

Abstract

Microscope à feuille de lumière, comprenant une chambre à échantillons dans laquelle un verre de couvercle ou de support peut être disposé, qui a une surface définissant une interface à réflexion partielle ; un système optique comportant un objectif tourné vers le verre de couvercle ou de support ; un système d'illumination conçu pour produire une feuille de lumière ; un capteur ; et un processeur. Le microscope à feuille de lumière forme un dispositif de mesure pour déterminer une grandeur de mesure. Le dispositif de mesure est conçu pour diriger la feuille de lumière à travers le système optique sous incidence oblique sur le verre de couvercle ou de support, pour produire un faisceau de lumière de réflexion en réfléchissant la feuille de lumière en partie au niveau de l'interface, et pour recevoir le faisceau de lumière de réflexion à travers le système optique et le diriger sur le capteur. Le capteur est conçu pour détecter l'intensité et/ou l'emplacement de l'incidence du faisceau de lumière de réflexion. Le processeur est conçu pour déterminer la grandeur de mesure sur la base de l'intensité détectée et/ou de l'emplacement d'incidence du faisceau de lumière de réflexion.
EP20719335.0A 2019-04-12 2020-03-25 Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons Pending EP3953684A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102019109832.7A DE102019109832B3 (de) 2019-04-12 2019-04-12 Lichtblattmikroskop und Verfahren zum Erfassen einer Messgröße
PCT/EP2020/058256 WO2020207795A1 (fr) 2019-04-12 2020-03-25 Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons

Publications (1)

Publication Number Publication Date
EP3953684A1 true EP3953684A1 (fr) 2022-02-16

Family

ID=70289729

Family Applications (1)

Application Number Title Priority Date Filing Date
EP20719335.0A Pending EP3953684A1 (fr) 2019-04-12 2020-03-25 Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons

Country Status (6)

Country Link
US (1) US11500189B2 (fr)
EP (1) EP3953684A1 (fr)
JP (1) JP7411682B2 (fr)
CN (1) CN113677979B (fr)
DE (1) DE102019109832B3 (fr)
WO (1) WO2020207795A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102020209268B3 (de) 2020-07-22 2021-10-14 Hochschule Emden/Leer Optisches System
CN113484297B (zh) * 2021-09-07 2021-12-24 清华大学 一种荧光光片显微系统

Family Cites Families (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS587986Y2 (ja) * 1977-03-22 1983-02-12 株式会社ユアサコーポレーション 電池
US6731380B2 (en) * 2001-06-18 2004-05-04 Applied Optics Center Of Delaware, Inc. Method and apparatus for simultaneous measurement of the refractive index and thickness of thin films
JP4242767B2 (ja) * 2001-09-21 2009-03-25 ケイマック ２次元型検出器を用いた薄膜特性測定装置及びその測定方法
EP1840623B1 (fr) * 2006-03-31 2013-05-08 Yokogawa Electric Corporation Microscope avec un système optique pour detecter un erreur de focalisation
DE102007043937B4 (de) 2006-09-13 2010-10-07 Innovent E.V. Verfahren zur Bestimmung der Dicke und des Brechungsindex von optisch transparenten Schichten auf optisch transparenten planparallelen Substraten
GB0814039D0 (en) 2008-07-31 2008-09-10 Imp Innovations Ltd Optical arrangement for oblique plane microscopy
WO2010135323A1 (fr) * 2009-05-19 2010-11-25 Bionanomatrix, Inc. Dispositifs et procédés pour la détermination dynamique de l'orientation spatiale d'un échantillon et le repositionnement dynamique d'un échantillon
TWI448055B (zh) 2010-06-07 2014-08-01 Richtek Technology Corp 切換式電源供應器之控制電路及其控制方法以及用於其中之電晶體元件
DE102010030430B4 (de) 2010-06-23 2015-01-29 Leica Microsystems Cms Gmbh Triangulierende Autofokuseinrichtung für Mikroskope und Verwendungen hiervon
DE102011000835C5 (de) * 2011-02-21 2019-08-22 Leica Microsystems Cms Gmbh Abtastmikroskop und Verfahren zur lichtmikroskopischen Abbildung eines Objektes
DE102011084562B4 (de) 2011-10-14 2018-02-15 Leica Microsystems Cms Gmbh Verfahren und Vorrichtung zur Feststellung und Korrektur von sphärischen Abbildungsfehlern in einem mikroskopischen Abbildungsstrahlengang
US20150330776A1 (en) * 2012-10-15 2015-11-19 Sony Corporation Image obtaining apparatus, and method of measuring an inclination of a slide
WO2015027153A1 (fr) 2013-08-22 2015-02-26 Thorlabs, Inc. Appareil de mise au point automatique
DE102013112595A1 (de) * 2013-11-15 2015-05-21 Carl Zeiss Microscopy Gmbh Anordnung zur Lichtblattmikroskopie
JP7180964B2 (ja) 2014-01-17 2022-11-30 ザ・トラスティーズ・オブ・コロンビア・ユニバーシティ・イン・ザ・シティ・オブ・ニューヨーク 三次元イメージング装置および方法
DE102014216227B4 (de) * 2014-08-14 2020-06-18 Carl Zeiss Microscopy Gmbh Verfahren und Vorrichtung zum Bestimmen eines Abstandes zweier voneinander entlang einer ersten Richtung beabstandeter optischer Grenzflächen
US20160187633A1 (en) * 2014-12-24 2016-06-30 Applied Scientific Instrumentation Inc. Dual oblique view single plane illumination microscope
DE102015221044A1 (de) * 2015-10-28 2017-05-04 Carl Zeiss Microscopy Gmbh Probenbegrenzungselement, Mikroskopierverfahren und Mikroskop
US11243391B2 (en) 2016-05-30 2022-02-08 The Trustees Of Columbia University In The City Of New York Three-dimensional imaging using swept confocally aligned planar excitation with asymmetrical magnification
DE102016212020A1 (de) * 2016-07-01 2018-01-04 Carl Zeiss Microscopy Gmbh Anordnung zur Mikroskopie und zur Korrektur von Aberrationen
DE102016212019A1 (de) 2016-07-01 2018-01-04 Carl Zeiss Microscopy Gmbh Neigungsmessung und -korrektur des Deckglases im Strahlengang eines Mikroskops
DE102016119268B3 (de) 2016-10-10 2017-12-21 Leica Microsystems Cms Gmbh Schiefebenenmikroskop
DE102017217380A1 (de) * 2017-09-29 2019-04-04 Carl Zeiss Microscopy Gmbh Immersionsvorrichtung zur dynamischen Anpassung eines Mediums an eine Probe

Also Published As

Publication number Publication date
CN113677979A (zh) 2021-11-19
JP7411682B2 (ja) 2024-01-11
DE102019109832B3 (de) 2020-04-23
CN113677979B (zh) 2024-07-02
WO2020207795A1 (fr) 2020-10-15
JP2022527609A (ja) 2022-06-02
US11500189B2 (en) 2022-11-15
US20220206281A1 (en) 2022-06-30

Similar Documents

Publication Publication Date Title
EP2137488B1 (fr) Procédé et dispositif de reproduction optique à discrimination en profondeur
EP3489735B1 (fr) Procédé et système de microscopie à feuille de lumière
EP2406679B1 (fr) Procédé d'autofocus et système d'autofocus
DE102006050834A1 (de) Grabenmesssystem mit einem chromatischen konfokalen Höhensensor und einem Mikroskop
DE102011082756A1 (de) Autofokussierverfahren und -einrichtung für ein Mikroskop
EP3948392B1 (fr) Procédé et dispositif de détection de déplacements d'un échantillon par rapport à un objectif
WO2001088590A1 (fr) Dispositif de mise au point automatique confocale
EP3953684A1 (fr) Microscope à feuille de lumière et procédé de détermination de l'indice de réfraction d'objets dans la chambre à échantillons
WO2016193037A1 (fr) Procédé pour la détermination d'une information de hauteur à résolution spatiale d'un échantillon au moyen d'un microscope à champ large et microscope à champ large
EP3374755A1 (fr) Microscope optique et procédé de détermination d'un indice de réfraction, dépendant de la longeur d'onde, d'un milieu échantillon
DE102009012293A1 (de) Autofokusverfahren und Autofokuseinrichtung
DE102019102330C5 (de) Optisches System für ein Mikroskop, Mikroskop mit einem optischen System und Verfahren zur Abbildung eines Objekts unter Verwendung eines Mikroskops
EP3418789A1 (fr) Procédé et système de microscopie destiné à enregistrer une image
WO2014147257A1 (fr) Procédé par microscopie optique de localisation d'objets ponctuels
EP3867684B1 (fr) Procédé et microscope pour déterminer l'épaisseur d'une lamelle couvre-objet ou porte-objet
EP3198323B1 (fr) Dispositif de projection d'un échantillon
DE102022202778A1 (de) System und Verfahren zur konfokal-chromatischen Linienabstandsmessung
EP3867630B1 (fr) Procede et microscope pour la determination de l'indice de refraction d'un milieu optique
EP2767797B1 (fr) Interféromètre à faible cohérence et procédé de mesure optique à résolution spatiale du profil de surface d'un objet
DE102018125995A1 (de) Verfahren und Mikroskop zur Bestimmung einer Verkippung eines Deckglases
DE102016120312B3 (de) Verfahren zum Beleuchten von Fokuspositionen objektseitig eines Objektivs eines Mikroskops und Mikroskop
DE102019116114B3 (de) Mikroskop und Verfahren zum Betrieb eines Mikroskops
DE202009003288U1 (de) Autofokuseinrichtung
DE102012109726A1 (de) Verfahren und Vorrichtung zur Bestimmung der Geometrie eines Objektes mit einer Zoomoptik
DE102018117470A1 (de) Verfahren zur Bestimmung von Dicke und Brechzahl einer Schicht

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20211022

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20221027

P01 Opt-out of the competence of the unified patent court (upc) registered

Effective date: 20230414