WO2019011581A1 - Scintillement lors d'un éclairage à angle variable - Google Patents

Scintillement lors d'un éclairage à angle variable Download PDF

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Publication number
WO2019011581A1
WO2019011581A1 PCT/EP2018/065945 EP2018065945W WO2019011581A1 WO 2019011581 A1 WO2019011581 A1 WO 2019011581A1 EP 2018065945 W EP2018065945 W EP 2018065945W WO 2019011581 A1 WO2019011581 A1 WO 2019011581A1
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WIPO (PCT)
Prior art keywords
illumination
light
light sources
geometries
optical device
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Application number
PCT/EP2018/065945
Other languages
German (de)
English (en)
Inventor
Lars STOPPE
Thomas OHRT
Original Assignee
Carl Zeiss Microscopy 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 Carl Zeiss Microscopy Gmbh filed Critical Carl Zeiss Microscopy Gmbh
Priority to CN201880052636.7A priority Critical patent/CN110998406B/zh
Publication of WO2019011581A1 publication Critical patent/WO2019011581A1/fr

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Classifications

    • 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
    • G02B21/06—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
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/14—Condensers affording illumination for phase-contrast observation
    • 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

Definitions

  • Various examples generally relate to illuminating a
  • Sample object by means of multiple illumination geometries which implement an angle-variable illumination.
  • various examples relate to illuminating the sample object or an environment of the sample object by means of continuous illumination.
  • flicker can be reduced due to the lighting by means of the multiple lighting geometries.
  • Illumination of a sample object used in imaging the light intensity varies as a function of the angle of incidence. Due to the angle-variable illumination different illumination geometries can be implemented. For example, different
  • Illuminating geometries illuminate a specimen object from different angles or areas, which can be implemented by a spatial structure of the light sources of a lighting module.
  • the angle-variable illumination by means of different illumination geometries may be desirable in connection with different applications.
  • phase contrast See, for example, DE 10 2014 1 12 242 A1 or, for example, L. Tian and L. Waller: "Quantitative differential phase contrast imaging in an LED array microscope", Optics Express 23 (2015), 1 1394.
  • angle variable illumination involves the creation a height profile in the material testing, see, for example, German Patent Application 10 2017 106 984.4.
  • Lighting geometries are perceived by a user as “flickering.”
  • the brightness or light distribution typically changes from
  • Illumination geometry to illumination geometry may have a frequency in the range of 10 Hz to 200 Hz. Such a flicker can be perceived by the user as unpleasant.
  • an optical device controller is configured to drive a plurality of first light sources of the optical device for sequentially illuminating a sample object disposed on a sample holder of the optical device by a plurality of illumination geometries.
  • the controller is also set up to have at least one second light source for
  • the use of the continuous lighting may cause the perception of flickering with the sequential lighting by means of the plurality of illumination geometries is reduced.
  • the continuous illumination can provide a comparatively large basic brightness value, so that a modulation of this basic brightness value due to the sequential illumination by means of the plurality of illumination geometries does not have a strong influence on the brightness perception of a user.
  • One method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source to illuminate the sample object or an environment of the sample object by means of a continuous illumination during the
  • a computer program product includes program code that can be executed by at least one processor. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source to illuminate the sample object or an environment of the sample object by means of a continuous illumination during illumination by means of the plurality of illumination geometries.
  • a computer program includes program code that comes from at least one
  • Processor can be executed. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source for Illumination of the sample object or an environment of the sample object by means of a continuous illumination during illumination by means of the plurality of illumination geometries.
  • an optical device controller is configured to drive a plurality of light sources of the optical device for sequentially illuminating a sample object disposed on a sample holder of the optical device by a plurality of illumination geometries. At least a subset of the plurality of light sources operates at at least two illumination geometries of the plurality of illumination geometries at at least one light intensity other than zero.
  • the luminous intensity of the light sources of the subset is not set to zero when switching between different illumination geometries. This in turn creates a basic brightness value that allows the perception of flicker to be reduced.
  • a method comprises driving a plurality of light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of
  • Lighting geometries At least a subset - or all - of the plurality of light sources is operated at at least two illumination geometries of the plurality of illumination geometries at at least one light intensity other than zero.
  • the plurality of light sources is arranged in the bright field of a detector aperture of a detection optical system of the optical device.
  • a computer program product includes program code that can be executed by at least one processor. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries. At least a subset of the plurality of light sources will be at least two Operated illumination geometries of the plurality of illumination geometries at least one light intensity, which is different from zero.
  • the plurality of light sources is the bright field of a detector aperture of a detection optical system of the optical
  • a computer program includes program code that comes from at least one
  • Processor can be executed. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries. At least a subset of the plurality of light sources is operated at at least two illumination geometries of the plurality of illumination geometries at at least one light intensity other than zero.
  • the plurality of light sources is the bright field of a detector aperture of a detection optical system of the optical
  • One method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source to illuminate the sample object or an environment of the sample object by means of continuous-wave illumination. It can the
  • CW lighting means continuous lighting in comparison to sequential lighting.
  • a computer program product includes program code that can be executed by at least one processor. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source for Illumination of the sample object or an environment of the sample object by means of a continuous-wave illumination.
  • a computer program includes program code that comes from at least one
  • Processor can be executed. Executing the program code causes the at least one processor to perform a method.
  • the method comprises driving a plurality of first light sources of an optical device for sequentially illuminating a sample object arranged on a sample holder of the optical device by means of a plurality of illumination geometries.
  • the method also includes driving at least one second light source to illuminate the sample object or an environment of the sample object by means of continuous-wave illumination.
  • FIG. 1 schematically illustrates an optical device and a controller for the optical device according to various examples.
  • FIG. 2 schematically illustrates a lighting module of the optical device according to various examples, which has a plurality of light sources in a lattice structure.
  • FIG. 3 schematically illustrates the light intensity with which different light sources of the illumination module are operated in an exemplary illumination geometry.
  • FIG. 4 schematically illustrates illumination geometries according to FIG. 4
  • FIG. 5 schematically illustrates illumination geometries according to various examples.
  • FIG. 6 schematically illustrates illumination geometries according to various examples.
  • FIG. 7 schematically illustrates brightness values for the illumination geometries according to the example of FIG. 5th
  • FIG. 8 schematically illustrates brightness values for the illumination geometries according to the example of FIG. 6th
  • FIG. 9 schematically illustrates illumination geometries and associated ones
  • FIG. 10 schematically illustrates illumination geometries and associated ones
  • FIG. 1 1 schematically illustrates illumination geometries according to FIG. 1
  • FIG. 12 schematically illustrates illumination geometries according to various examples.
  • FIG. 13 schematically illustrates a lighting module and a sample holder according to various examples.
  • FIG. 14 schematically illustrates a lighting module and a sample holder according to various examples.
  • FIG. 15 schematically illustrates the spectral separation of light according to FIG.
  • FIG. 16 is a flowchart of an example method
  • FIG. 17 is a flowchart of an example method.
  • Units can be implemented as hardware, software or a combination of hardware and software.
  • An illumination geometry may be characterized by the amount of angles or directions under which the sample object is illuminated.
  • illumination geometry may refer to the illumination of the sample object in the bright field, i. those angles or directions in the bright field under which the sample object is illuminated.
  • illumination geometries can be implemented in the various techniques described herein.
  • lighting geometries could be implemented that provide semicircular or linear illumination of the sample object.
  • Lighting geometries could be used which illuminate the sample object from different illumination directions, wherein the different illumination directions can be mirrored, for example, on the optical axis. It could also be used such illumination directions, which are mirrored on a symmetry axis oriented perpendicular to the optical axis - as it may be the case, for example, in complementary semicircular or linear illumination geometries.
  • the techniques described herein may be applications
  • Sample object to be a biological sample could be arranged on a sample holder of the microscope.
  • the techniques described herein could also be used in conjunction with materials testing.
  • a height profile of a sample object - for example a textile fabric - could be checked in incident light geometry.
  • Lighting geometries according to reference implementations can cause an unpleasant flicker. Namely, when switching between different lighting geometries, the lighting of the surrounding scene typically changes. The brightness values can fluctuate. Often a such flicker frequencies in the range of 10 Hz - 200 Hz, which should be avoided in the context of epilepsy.
  • this is enabled by providing a basic brightness value.
  • This basic brightness value is then modulated by switching between different illumination geometries, wherein the amplitude of the modulation, for example, may be comparatively small compared to the amplitude of the basic brightness value, for example not> 50%, optionally not more than 15%, further optionally not larger than 5%. This will increase the perception of the
  • the basic brightness value can be created, for example, by the use of continuous illumination - sometimes referred to as continuous-wave illumination.
  • continuous illumination may mean that a possible frequency with which the continuous illumination is switched on and off is significantly lower than the frequency with which switching between different illumination geometries of the angle-variable illumination.
  • a frequency with which the continuous illumination is switched on and off can not be greater than 5% of the frequency with which between different illumination geometries of the angle variable
  • Lighting is switched, its optional not greater than 0.5%, further optional not greater than 0.05%.
  • the basic brightness value could be generated by one or more light sources located close to those light sources used to illuminate the sample object using the plurality of illumination geometries.
  • the one or more light sources used for the continuous illumination could, for example be integrated into the optical device.
  • these one or more light sources it would be possible for these one or more light sources to be integrated in a lighting module which is also suitable for illuminating the sample object by means of the plurality of light sources
  • Lighting geometries of angle-variable lighting is used.
  • an external light source could be used to provide the basic brightness value.
  • an external light source could be used as part of a projector, so that a projector could be used
  • Outer surface of the optical device can be illuminated.
  • Such light sources for continuous illumination can be arranged in a dark field of a detector aperture of a detection optical system of the optical device.
  • the basic brightness value could also be obtained by limiting the difference between the intensities of different light sources used in different illumination geometries. This means that the light sources are at least partially not switched off and on when switching between lighting geometries; rather, it would be possible for at least some of the light sources to switch between
  • Lighting geometries between different finite light intensities are switched back and forth. This limits the stroke of change in the brightness value and thus provides a basic brightness value that limits the perception of flicker.
  • the basic brightness value could be achieved by operating one or more light sources arranged in the bright field of the detector aperture at a constant light intensity.
  • a reference illumination geometry can be used to compensate for an impact of these constant-intensity light sources on the imaging.
  • Such different approaches to limiting the perception of flicker can be combined. Such techniques may make it possible to provide the optical device with an open and thus easily accessible sample holder and illumination module, without the use of several lighting geometries in the angle-variable lighting would have a negative impact on the working environment.
  • the flickering can be reduced particularly efficiently, in particular, if light of the same color is used to illuminate the sample object by means of the plurality of illumination geometries of the angle-variable illumination and to provide the basic brightness value.
  • This may mean that light is used which has no or only a comparatively small spectral distance, e.g. a spectral distance of not more than 100 nm, optionally not more than 50 nm, further optionally not more than 5 nm. Namely, a distinction of the various light sources by the user can be avoided, so that the flicker can be suppressed particularly efficient.
  • FIG. 1 illustrates an example optical system 100.
  • the optical system 100 according to the example of FIG. 1 a light microscope
  • the optical system 100 implement, for example, in transmitted-light geometry. Such a microscope could be used for phase-contrast imaging.
  • the optical system 100 according to the example of FIG. 1 also implement a light microscope, in Auflichtgeometrie.
  • a corresponding light microscope in Auflichtgeometrie could be used for material testing. For this a height profile of the sample object can be created.
  • optical system 100 By means of the optical system 100, it may be possible to enlarge small structures of a sample object fixed by a sample holder 13.
  • the optical system 100 could be a wide field microscope
  • the detection optics 12 may generate an image of the sample object on a detector 14.
  • the detector 14 may then be configured to capture one or more images of the sample object.
  • a viewing through an eyepiece is conceivable.
  • the detection optics 1 12 has a detector aperture.
  • the size of the detector aperture defines which light sources of a lighting module 1 1 1 are arranged in the bright field or in the dark field.
  • the illumination module 1 1 1 is arranged to illuminate the sample object which is fixed on the sample holder 1 13.
  • this lighting could be implemented by means of Köhler illumination.
  • a condenser lens and a condenser aperture diaphragm are used. This leads to a particularly homogeneous intensity distribution of the light used for the illumination in the plane of the sample object.
  • a partially incoherent illumination can be implemented.
  • the lighting module 1 1 1 is set up to enable angle-variable lighting. This means that by means of
  • Illumination module 1 1 1 different illumination geometries of the
  • Illumination of the sample object used light can be implemented.
  • the different illumination geometries can correspond to illumination of the sample object from different illumination directions.
  • different hardware implementations are possible to accommodate the different ones
  • Illumination module 1 1 1 comprise a plurality of adjustable light sources, which are adapted to locally modify and / or generate light.
  • the controller 1 15 could be implemented as a microprocessor or microcontroller. Alternatively or additionally, the controller 1 15 could include, for example, an FPGA or ASIC. The controller 1 15 may alternatively or additionally also the sample holder 1 13, the imaging optics 1 12, and / or the detector 1 14 control. In some examples, it is possible that the controller 1 15 in a housing of the optical device 100 is integrated. In other examples, it would also be possible that the
  • Control 1 15 is provided externally of the optical device 100.
  • controller 1 15 by a corresponding
  • FIG. 2 illustrates aspects relating to the lighting module 1 1 1.
  • the lighting module 1 1 1 a variety of adjustable
  • Light sources 121 -1, 121 -2 in a matrix structure (lattice structure with square unit cell).
  • the matrix structure is oriented in a plane perpendicular to the beam path of the light (lateral plane, spatial coordinates x, y). This plane is turned toward the sample holder (the sample holder is not shown in FIG.
  • Light sources 121 -1, 121 -2 so be arranged in a lattice structure.
  • the adjustable light sources 121 -1, 121 -2 could be implemented as lights, for example as light emitting diodes.
  • the adjustable light sources 121 -1, 121 -2 could be implemented as lights, for example as light emitting diodes.
  • Lighting geometry can be implemented.
  • the lighting module 1 1 1 could be implemented as a spatial light modulator (SLM).
  • SLM spatial light modulator
  • the SLM may be spatially resolved to engage in a condenser pupil, which may have a direct impact on imaging - for example, modeled using the TCC.
  • the different light sources e.g. by micromirrors or switchable
  • the double detector aperture 399 of the detection optical system 1 14 is also shown.
  • FIG. 3 illustrates aspects related to an exemplary lighting geometry 300.
  • the light intensity 301 provided for the various adjustable light sources 121 of the lighting module 1 1 1 along the axis XX ' of FIG. 2 shown.
  • the illumination system 300 is characterized by a dependency of
  • Illumination geometry 300 provided.
  • the illumination geometry 300 is typically characterized by the light intensity 301 of the light sources 121-1 forming the bright field illumination.
  • FIG. Figure 4 illustrates aspects relating to complementary illumination geometries 300-1, 300-2 that may be used in conjunction with the angle variable illumination.
  • FIG. 4 illustrates a scenario in which in each case only parts of the light sources 121 -1 of the illumination modules 1 1 1 contribute to the illumination by means of the illumination geometries 300 - 1, 300 - 2. Same hatchings denote the same light intensities. In the example of FIG. 4 a big flicker was observed. This is because the illumination geometry 300-1 is fully complementary to the
  • Illumination geometry is 300-2.
  • Illumination geometry 300-1 as well as during illumination of the
  • Sample object by means of the illumination geometry 300-2 are turned on.
  • FIG. Figure 5 illustrates aspects relating to illumination geometries 300-1, 300-2 that may be used in conjunction with the angle-variable illumination.
  • the illumination geometries 300-1, 300-2 are in the scenarios of FIG. 4 and FIG. 5 identical, because in the bright field, the same semicircular pattern for
  • FIG. 5 illustrates in particular aspects relating to the flicker reduction by means of a continuous illumination by light sources 121 -2 arranged in the dark field.
  • FIG. 5 While the light sources 121 -1 convert the illumination geometries 300-1, 300-2 in the bright field, FIG. 5 also uses light sources 121-2 in the dark field in order to provide a basic brightness value by means of continuous illumination. This reduces the flicker.
  • those light sources 121 - 2 arranged in the dark field are switched on continuously during illumination with the illumination geometries 300 - 1, 300 - 2 and thus provide the constant basic brightness value. This means that these are in the dark field of the
  • Detector aperture arranged light sources 121 -2 of the lighting module 1 1 1 not or only comparatively slowly on and off.
  • These light sources 121 -2 arranged in the dark field of the detector aperture implement the continuous illumination.
  • Lighting module 1 1 1 used both for the illumination of the sample object by means of the variety of illumination geometries of the angle-variable illumination, as well as for implementing the continuous illumination.
  • the light sources 121 - 1 of the lighting module 1 1 which provide the illumination of the sample object by means of the plurality of lighting geometries of the angle variable lighting, as well as the light sources 121 - 2 of the
  • Illumination module 1 1 which is the continuous illumination of the sample object (see Figure 2): for example, the light sources 121-2 could be disposed in one or more outer rings of a ring lattice structure, while the lattice structure could be arranged in a common lattice structure on a corresponding sample holder surface
  • Light sources 121 -1 could be arranged in one or more inner rings of a ring-grid structure. In this case, there is a spatial separation of the light of the various light sources 121 -1, 121 -2 through the detector aperture 399 after the incident light on the sample object.
  • a conventional lighting module such as a lighting module as it is known from German Patent Application 10 2016 1 16 31 1 .2.
  • the flicker can be reduced particularly efficiently because the light sources providing the continuous illumination are located close to the light sources used to implement the different illumination geometries. This means that essentially the same areas through the
  • the illumination geometries 300-1, 300-2 according to the
  • Example of FIG. 5 can be used to generate a result image which images the illuminated sample object with a phase contrast.
  • 300-1, 300-2 could each have a picture of the
  • Sample object can be detected by the detector 1 14, and then based on these two images, a result image can be determined.
  • the result image can be determined by combining the two images, such as the following
  • FIG. Figure 6 illustrates aspects relating to illumination geometries 300-3, 300-4 that may be used in conjunction with the angle variable illumination.
  • FIG. 6 basically corresponds to the example of FIG. 5th
  • FIG. 6 illustrates in particular aspects relating to the flicker reduction by means of a continuous illumination by light sources 121 -2 arranged in the dark field.
  • the illumination geometries 300-3, 300-4 according to the
  • Example of FIG. 6 can be used for an autofocus application. For this, a shift of the image of the sample object between images viewed in the illumination geometry 300-3 and the illumination geometry 300-4 can be considered. This shift can be proportional to the defocusing. From the knowledge of the distance of the lines of the illumination geometries 300-3, 300-4 from each other or from the optical axis, the defocusing can also be determined quantitatively. Corresponding techniques are e.g. in DE10 2014 109 687 A1.
  • FIG. 7 shows the basic brightness value 181, which in the scenario of FIG. 5 by the continuous lighting by means of Light sources 121 -2, which are located outside the double detector aperture 399, is achieved.
  • the brightness value 182 indicated by the semicircular illumination geometries of the light sources 121-1 disposed within the double detector aperture 399, according to the scenario of FIG. 5 is achieved.
  • the modulation of the basic brightness value 181 by switching between the illumination geometries 300-1, 300-2 amounts to approximately 25%.
  • the light sources 121-2 providing the continuous illumination could be driven such that such a ratio between the brightness values 181, 182 is achieved.
  • the basic brightness value 181 could be based on the brightness value 182 of the various
  • Illumination geometries 300-1, 300-2 are implemented. It could be for
  • a corresponding control loop can be implemented that has a
  • Brightness sensor such as a photodiode, and takes into account the ratio between the brightness values 181, 182 as a target size and the light intensity at which each of the light sources 121 -2 -2 emits light, and / or the number of light sources 121 -2 as a manipulated variable considered.
  • the brightness value 182 could also be determined by a given control table, e.g. depending on the illumination geometry used and / or the light intensities with which the various light sources 121 -1 emit light.
  • the brightness value 182 in FIG. 8 is absolutely smaller than the brightness value 182 in FIG. 7, because in the scenario of FIG. 6, a linear illumination of the light sources 121-1 is used as illumination geometries 300-3, 300-4, in the scenario of FIG. 5 but a semi-circular illumination of the light sources 121 -1 is used as lighting geometries 300-1, 300-2.
  • Exposure of the sample to be reduced with light Bleaching or phototoxic effects can be avoided. At the same time, flickering can be efficiently reduced.
  • an ambient brightness could also be considered, i. the brightness value in the environment, which is not due to the illumination by means of the illumination geometries.
  • a sensor may be provided which is arranged on the optical device 100, for example on the sample holder, on the stand or in the vicinity of the illumination module 11. This is based on the
  • FIG. Figure 9 illustrates aspects relating to illumination geometries 300-5, 300-6 that may be used in conjunction with the angle-variable illumination.
  • semi-circular illumination geometries 300-5, 300-6 are also used, with light sources 121 -1 dimmed in bright field to provide a basic brightness value.
  • FIG. 9. Illustrates therefore in particular aspects relating to the flicker reduction by means of a modulation of the light intensity of light sources arranged in the bright field 121 -1 between light intensities greater than zero.
  • the different light sources 121 -1 are operated in the bright field in both illumination geometries 300-5, 300-6 with a light intensity that is greater than zero. For example, at the
  • Illumination geometry 300-5 operated in the left semicircle light sources 121 -1 at a first light intensity (shadows from bottom left to top right) and located in the right semicircle light sources 121 -1 operated at a second light intensity (hatching from top left to bottom right ); on the other hand, in the illumination geometry 300-6, the light sources 121-1 located in the left semicircle are operated at the second light intensity and the light sources 121-1 located in the right semicircle are operated at the first light intensity. In general, four different levels of light could be used. The local Variation of the brightness values 182, 183 is then comparatively small when switching between the different illumination geometries 300-5, 300-6 and so that we reduce the perception of the flicker. For example, the result image with phase contrast could be determined in this example: wherein the ratio between the brightness values 182, 183 and 0 ⁇ x ⁇ 1 applies.
  • FIG. 10 basically corresponds to FIG. 9, wherein in FIG. 10 more
  • Illumination geometry 300-7 is used.
  • An image of the sample object can also be detected for the illumination geometries 300-7.
  • the result image with phase contrast can be determined according to the following equation:
  • ⁇ denotes the brightness value 184, which is associated with the illumination geometry 300-7.
  • the illumination geometry 300-7 serves as a reference illumination geometry, wherein 11 and 12 are corrected by subtraction with ab. Thereby differential images - a ⁇ I 3 and I 2 - a - 1 3 are obtained.
  • the light sources 121-2 could be used to provide continuous illumination (not illustrated in Figs. 9 and 10).
  • 100% of all light sources 121 -1 are operated in the bright field in all illumination geometries 300-5, 300-6, 300-7 at light intensities greater than zero. Therefore, the flicker is reduced particularly well. It is not always necessary that 100% of all light sources 121 -1 be operated in the bright field continuously at a light intensity of greater than zero in order to achieve a reduction of the flicker. This is illustrated in conjunction with the examples of FIGS. 11 and 12.
  • FIG. Fig. 11 illustrates aspects relating to illumination geometries 300-3, 300-4 that may be used in conjunction with the angle variable illumination.
  • FIG. 1 two line-shaped illumination geometries 300-3, 300-4 are used, wherein the illumination lines have equal distances to the optical axis (in the center of the detector aperture 399 and perpendicular to the drawing plane, but not shown).
  • Such illumination geometries can be used in particular in the
  • illumination geometries 300-3, 300-4 shown in FIG. 1 an increased flicker was also observed.
  • illumination geometries 300-8-300-10 according to the example of FIG. 12 are used.
  • Illumination provided by the light sources 121 -2 (cf., FIG. 6).
  • FIG. Figure 12 illustrates aspects relating to illumination geometries 300-8, 300-9, 300-10 that may be used in conjunction with the angle-variable illumination.
  • an illumination geometry 300-8 is used that is in the bright field inverse to the illumination geometry 300-3 according to the example of FIG. 1 is 1; Accordingly, in the example of FIG. 12 also uses a lighting geometry 300-9, which in the bright field inverse to the
  • Illumination geometry 300-4 according to the example of FIG. 1 is 1.
  • a subset of the plurality of light sources 121-1 in the bright field of the detector aperture 399 is operated on at least two of the plurality of illumination geometries used at a constant light intensity other than zero.
  • all the light sources 121 -1 in the bright field, not on one of the two Lighting lines fall, operated at a constant light intensity. This creates the basic brightness value that reduces the perception of flicker.
  • Difference image are obtained, which corresponds to the illumination of the sample object by means of the line-shaped illumination geometry 300-3. Accordingly, a difference image can be obtained by subtracting a picture acquired with illumination by means of the reference illumination geometry 300 - 10 and an image acquired with illumination by means of the illumination geometry 300 - 9, which indicates the illumination of the sample object by means of the linear image
  • Illumination geometry corresponds to 300-4. Such pre-processing of the images can then be established based on corresponding difference images
  • the subset of the light sources 121 -1 in the bright field which have a constant light intensity different from zero at the different illumination geometries and thus provide a basic brightness value, is dimensioned comparatively large.
  • the subset could not be less than 50% of all light sources within the
  • Brightfields optionally not less than 75%, further optional not less than 90%, further optional not less than 95%, further optional not less than 100%.
  • these sub-set light sources be operated at the constant light intensity for all lighting geometries used.
  • the basic brightness value for example, by driving the Be implemented light sources 121 -2 in the dark field or by suitable design of the illumination geometries, which are characterized by the used light intensities of the light sources 121 -1 in the bright field.
  • light sources could be used which are spatially separate from the light sources 100 a 20-1, 121 -2
  • FIG. 13 illustrates aspects relating to a lighting module 11 1.
  • FIG. 13 shows a scenario in which light sources 126, which illuminate an environment of the sample object by means of a continuous illumination, are provided on a side surface of the illumination module 11. Thereby, the basic brightness value can be provided so that flickering can be reduced.
  • the light sources 121 which are used to illuminate a sample object arranged on the sample holder 13, are located on one of the light sources 121, which are used to illuminate a sample object arranged on the sample holder 13, are located on one of the light sources 121, which are used to illuminate a sample object arranged on the sample holder 13, are located on one of the light sources 121, which are used to illuminate a sample object arranged on the sample holder 13, are located on one of the
  • Sample holder facing surface of the lighting module 1 1 1 - and thus spatially separated from the light sources 126 - are arranged.
  • the orientation and / or arrangement of the light sources 121, 126 we achieve a spatial separation of the corresponding light, in particular before the light of the light sources 126 impinges on the sample object. Thereby, exposure of the sample object to light from the light sources 126 can be avoided.
  • FIG. 14 illustrates aspects relating to a sample holder 1 13.
  • FIG. 13 illustrates a scenario in which the light sources 126 are provided which surround an environment of the sample object by means of continuous illumination illuminate. Thereby, the basic brightness value can be provided so that flicker can be reduced.
  • the light sources 126 are arranged on the sample holder 123.
  • the light sources 126 are configured to emit light away from a sample object disposed on the sample holder.
  • the arrangement and / or orientation of the light sources 126 results in a spatial separation between light emitted by the light sources 121 and by the light sources 126
  • the light sources 126 were provided as part of the optical device 100. In other implementations, it would also be possible for the light sources configured to provide the light for continuous illumination not to be located in the optical device 100, but separately. For example, it would be possible to use directional continuous illumination of suitable light sources, for example one
  • one or more objects could be selectively illuminated around the optical device to provide a basic brightness value that reduces flicker. Exposure of the sample to the light is avoided by the directional nature of the light source. For example, an outer surface of the optical device could be illuminated. For example, surfaces of the tripod of the optical device could be illuminated. As a result, a homogenization of the brightness can be achieved.
  • micro-projectors could be used, which several Have light sources with adjustable colors. Then it would be possible for the color of the light emitted by the micro-projectors to be matched to the color of the light used to illuminate the sample object by means of the different illumination geometries.
  • the spatial separation can be implemented by a detector aperture, the further light being emitted by light sources in the dark field of the detector aperture.
  • Other examples described above allow for spatial separation by the orientation and / or arrangement of the light sources.
  • light sources can be used, which are arranged on different surfaces of the lighting module.
  • An external light source could also be used to provide the further light, for example in the context of a projector. In some examples, however, it would be possible to dispense with such a spatial separation of light. This is in
  • FIG. 15 shows the spectrum of light 621 emitted by light sources 121 on a lighting module 1 1 1.
  • These light sources 121 on the lighting module 1 1 1 also send out light 622.
  • double-pixel light emitting diodes could be used. This means that there is no spatial separation between the light 621 and the light 622.
  • the frequency offset could correspond to a wavelength difference of not more than 5 nm, optionally not more than 2 nm, more optionally not more than 1 nm.
  • the light 622 may implement the continuous illumination and the light 621 may be used to implement different illumination geometries. Through a bandpass filter can then be prevented that the light 622 falls on the detector 1 14. At the same time, however, the light 622 can radiate into an environment of the optical device 100, so that a basic brightness value is created by means of the light 622, which serves to reduce the flicker.
  • the passband 652 of a corresponding bandpass filter is shown. This achieves the spectral separation.
  • FIG. 16 is a flowchart of an example method.
  • a plurality of first light sources are driven in order to illuminate a sample object by means of a plurality of illumination geometries.
  • an angle variable illumination of the sample object can be provided.
  • At least one second light source is driven to the
  • the continuous illumination To illuminate the sample object or an environment of the sample object by means of a continuous illumination. This enables the creation of a basic brightness value, so that the flickering associated with the switching between the different illumination geometries is perceived comparatively weakly.
  • the continuous illumination it would be possible for the continuous illumination to be implemented by emitting light by means of the at least one second light source at a constant light intensity during the entire execution of block 1001.
  • the sample object is illuminated by means of the multiplicity of first light sources and the environment of the sample object is illuminated by means of the at least one second light source (and not the sample object itself), a spatial separation of the corresponding light can take place. It would also be possible for there to be a spatial separation of the light in which both the plurality of first light sources, as well as the at least one second light source illuminate the sample object, wherein, however, the at least one second light source is arranged in the dark field of a detector aperture of the detection optics. It would also be possible to perform a spectral separation of the light, for example by a bandpass filter.
  • FIG. 16 An exemplary implementation of the method according to FIG.
  • FIG. 16 is shown in conjunction with FIGS. 4-8.
  • FIG. 17 is a flowchart of an example method.
  • a multiplicity of light sources for illuminating a sample object are activated by means of a multiplicity of illumination geometries.
  • at least one subset-or all-of the plurality of light sources is driven in such a way that it emits light with at least one light intensity of greater than at the different illumination geometries - for example, a single constant light intensity can be used or several different ones
  • the plurality of light sources can be arranged in the bright field of a detector aperture of a detection optical system of the optical device.
  • a first illumination geometry of the plurality of illumination geometries may correspond to the operation of at least one first light source of the subset of the plurality of light sources at a first light intensity.
  • the first illumination geometry may also correspond to the operation of at least one second light source of the subset of the plurality of light sources at a second light intensity.
  • Illumination geometries may be the operation of the at least one first
  • Light source at a third light intensity and the operation of the at least one second light source at a fourth light intensity correspond.
  • each of the first light intensity, the second light intensity, the third light intensity and the fourth light intensity can be greater than zero.
  • the first light intensity may be different from the second light intensity, and the third light intensity may be different from the fourth light intensity.
  • the first light intensity it would be possible for the first light intensity to be equal to the fourth
  • the first light intensity to be different from the third light intensity and for the second light intensity to be different from the fourth light intensity
  • FIG. 16 An exemplary implementation of the method according to FIG. For example, FIG. 16 is shown in conjunction with FIGS. 9-12.
  • a detector could also be actuated in order to acquire an image of the sample object by means of detection optics for each illumination geometry. Based on these images, different applications could then be implemented: for example, a
  • Result image can be determined by combining the images, the result image has a phase contrast; it could also be the defocusing of the
  • Sample object along the optical axis can be determined, for example, by comparing the positions at which the sample object in the
  • light sources could be up on a sample holder
  • directed light cone can be used. It could also be used drowned bright field as a backlight source (see FIG 9). Techniques of inverse implementation of illumination geometries may also be used (see FIG. 10). Also, light patterns by means of light sources, which are arranged externally of the optical device, are possible, for example by means of
  • the techniques described herein may allow for flicker to be reduced flexibly for different detector apertures.
  • such techniques which do not rely on the use of light sources in the dark field of the detector aperture, can be used flexibly for different detector apertures.
  • it may also be possible to flexibly adjust the intensity of the flicker depending on the tolerable light exposure of the sample object.
  • the basic brightness value can be adjusted by suitably selecting the luminous intensity and / or the number of light sources contributing to the continuous illumination; the ratio between basic brightness value and modulation due to
  • Switching between different illumination geometries can in turn be a measure of the intensity of the flicker.
  • Various herein described Techniques can also be flexible in incident illumination or
  • Transmitted light illumination can be used.
  • Lighting geometries on the one hand and for continuous lighting on the other hand are based on combining techniques based on a spectral separation of the light.

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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)
  • Computer Vision & Pattern Recognition (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

On observe que, lors de l'utilisation de différentes géométries d'éclairage (300-1, 300-2) en lien avec l'éclairage à angle variable, on peut percevoir un scintillement dû à la commutation entre les géométries d'éclairage. La présente invention a pour but de réduire ce scintillement. Dans différents exemples, cela est obtenu par la génération d'une valeur de luminosité de base.
PCT/EP2018/065945 2017-07-12 2018-06-15 Scintillement lors d'un éclairage à angle variable WO2019011581A1 (fr)

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