WO2016157458A1 - Appareil de mesure, système de mesure, procédé de traitement de chaîne de signal, et programme - Google Patents

Appareil de mesure, système de mesure, procédé de traitement de chaîne de signal, et programme Download PDF

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Publication number
WO2016157458A1
WO2016157458A1 PCT/JP2015/060280 JP2015060280W WO2016157458A1 WO 2016157458 A1 WO2016157458 A1 WO 2016157458A1 JP 2015060280 W JP2015060280 W JP 2015060280W WO 2016157458 A1 WO2016157458 A1 WO 2016157458A1
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fluorescence
light
signal
signal sequence
pixel
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PCT/JP2015/060280
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English (en)
Japanese (ja)
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浜島　宗樹
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株式会社ニコン
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Priority to PCT/JP2015/060280 priority Critical patent/WO2016157458A1/fr
Publication of WO2016157458A1 publication Critical patent/WO2016157458A1/fr

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    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence

Definitions

  • the present invention relates to a measuring apparatus, a measuring system, a signal sequence processing method, and a program.
  • Patent Document 1 discloses a microscope apparatus including a dichroic mirror and an in-focus signal generator for providing an autofocus function (hereinafter referred to as “AF function”).
  • AF function an autofocus function
  • the two fluorescent lights having different wavelengths have different amounts of chromatic aberration, so that focusing is required every time an image is acquired.
  • the time required for focusing using the AF function is increased, which greatly affects the throughput of image acquisition in the imaging apparatus.
  • a light source device for irradiating an irradiated body with the first excitation light and the second excitation light, and a microlens array including a plurality of microlenses arranged two-dimensionally A sensor in which light receiving elements are two-dimensionally arranged, and the first fluorescence emitted when the first excitation light is irradiated onto the irradiated body and the second excitation light that is irradiated with the first excited light.
  • the first result is obtained from the sensor that receives the second fluorescence emitted when the light is irradiated through the microlens array and the signal sequence of the first pixel signal corresponding to the focal point of the first fluorescence.
  • a control unit that generates and generates a second result from the signal sequence of the second pixel signal corresponding to the focal point of the second fluorescence.
  • the measurement device described above a reaction device that reacts the irradiated object supported by a support member with a sample, and a transport device that transports the support member to the measurement device.
  • a measurement system is provided.
  • the first excitation light and the second excitation light are applied to the irradiated object by the step of disposing the support member on which the plurality of irradiated objects are disposed on the stage and the light source device.
  • a program for causing an information processing apparatus including at least a calculation unit and a storage unit to process a signal sequence of light received through a microlens array, the signal sequence of the light Is a set of pixel signals at a plurality of different focal positions, and the pixel signal acquired during the time zone in which the first fluorescence will be emitted and the time in which the second fluorescence will be emitted.
  • the figure explaining the extraction process of a pixel signal in case a microlens array is arrange
  • positioned at the imaging surface (Z 0) of an objective lens.
  • the figure explaining the extraction process of a pixel signal in case the image plane of an objective lens exists in the position (Z h1) shifted
  • the figure explaining the amount of chromatic aberration of fluorescence The figure which shows an example of the component of a control apparatus. An example of the flowchart at the time of measuring a to-be-irradiated body with the measuring apparatus which concerns on 1st Embodiment. An example of the detailed content of the flowchart of FIG. The figure explaining the procedure which determines a reference focus position. The figure explaining the procedure which determines a reference focus position. An example which acquires the signal sequence of the pixel signal corresponding to the focal point of the 1st fluorescence. An example which acquires the signal sequence of the pixel signal corresponding to the focal point of the 2nd fluorescence.
  • the other example of the flowchart at the time of measuring a to-be-irradiated body with a measuring apparatus The other example of the flowchart at the time of measuring a to-be-irradiated body with a measuring apparatus.
  • the figure explaining the procedure which determines a reference focus position. An example of acquiring a signal sequence of pixel signals corresponding to the focal point of the first fluorescence and a signal sequence of pixel signals corresponding to the focal point of the second fluorescence. It is another example of a structure of the measuring apparatus which concerns on 2nd Embodiment.
  • FIG. 1 An example of the flowchart at the time of measuring a to-be-irradiated body with the measuring apparatus which concerns on 3rd Embodiment.
  • the figure explaining the procedure which determines a reference focus position.
  • the figure explaining the relationship between the inclination of a to-be-irradiated body, and a focus position.
  • Another example of three points for determining the reference focal position The figure which shows the structure of the measurement system which concerns on 5th Embodiment.
  • an XYZ orthogonal coordinate system is set, and the positional relationship of each member will be described with reference to this XYZ orthogonal coordinate system.
  • the predetermined direction in the horizontal plane is the X-axis direction
  • the direction orthogonal to the X-axis direction in the horizontal plane is the Y-axis direction
  • the direction orthogonal to each of the X-axis direction and the Y-axis direction is the Z-axis direction.
  • the rotation (inclination) directions around the X axis, Y axis, and Z axis are the ⁇ X, ⁇ Y, and ⁇ Z directions, respectively.
  • FIG. 1 is a schematic configuration diagram illustrating an example of a measurement apparatus.
  • the measuring device is an optical microscope.
  • the measuring device may be another optical device or an imaging device.
  • the measurement device 10 includes a measurement device main body 20 that observes an object to be irradiated (measurement target) 1, a control device 30 that controls the operation of the measurement device main body 20, and a display device 40 that is connected to the control device 30.
  • the control device 30 includes a computer system.
  • the computer system includes at least a processor, a memory, and a storage device.
  • the memory is a volatile memory
  • the storage device is a nonvolatile storage such as a hard disk.
  • a computer system may include input devices such as a keyboard and a pointing device (eg, a mouse).
  • the display device 40 includes a flat panel display such as a liquid crystal display.
  • the measuring device main body 20 includes a light source device 2, an optical system 3, a stage 4, and a sensor 5.
  • the measuring apparatus main body 20 includes a body (not shown). Each of the light source device 2, the optical system 3, the stage 4, and the sensor 5 is supported by the body.
  • the light source device 2 can emit light having a plurality of different wavelengths.
  • the light source device 2 can emit excitation light to generate fluorescence from the irradiated object 1.
  • the light source device 2 can emit a plurality of excitation light beams having different wavelengths for generating fluorescence from the irradiated object 1 and reference light for obtaining reflected light from the irradiated object 1.
  • the light source unit 2 the light of wavelength ⁇ 1 as a first excitation light, light of the wavelength ⁇ 2 of the second excitation light, and the light of the wavelength lambda R of the reference light can be emitted.
  • the light source unit 2 the light of wavelength ⁇ 1 as a first excitation light, light of the wavelength ⁇ 2 of the second excitation light, and, in the wavelength lambda R of the reference light light, from the control unit 30 It is possible to selectively switch injection based on the signal.
  • the light source device 2 includes a light of wavelength ⁇ 1 as a first excitation light, are both capable of emitting a light of the wavelength .lambda.2, the wavelength lambda R of the reference light and the light of the second excitation light .
  • the wavelength ⁇ 1 and the wavelength ⁇ 2 and the wavelength lambda R is a different wavelength from each other.
  • the light source device 2 may emit third excitation light corresponding to the third fluorescence. Furthermore, the light source device 2 may emit the fourth excitation light corresponding to the fourth fluorescence.
  • a support member 60 in which a plurality of irradiated objects 1 are arranged is arranged on the stage 4.
  • the support member 60 is a plate.
  • the support member 60 is a plate-like member.
  • the irradiated object 1 is a biochip.
  • the biochip is sometimes called a microarray, a microarray chip, a biomolecule array, a biosensor, or the like.
  • FIG. 2 is an example of the support member 60 and the irradiated object 1.
  • the support member 60 is a glass plate. A plurality of biochips are fixed in a matrix on the glass plate. As an example, the biochip is bonded to the glass plate with an adhesive.
  • the biochip has a plurality of spots.
  • the biochip has a plurality of spots arranged in a matrix.
  • a biomolecule probe
  • different biomolecules are fixed to a plurality of spots on the biochip.
  • Each spot is set with an address so that the spot can be identified.
  • the address information is stored in the storage device of the control device 30, for example.
  • the irradiated object 1 emits fluorescence when irradiated with excitation light.
  • the irradiated object 1 generates fluorescence by irradiating the fluorescent dye bonded to the biomolecule of the irradiated object 1 with excitation light.
  • the irradiated object 1 generates the first fluorescence (wavelength: ⁇ 1 ′) when irradiated with the first excitation light (wavelength: ⁇ 1).
  • the irradiated object 1 generates second fluorescence (wavelength: ⁇ 2 ′) when irradiated with second excitation light (wavelength: ⁇ 2).
  • the wavelength ⁇ R of the reference light is not a wavelength for generating fluorescence from the irradiated object 1.
  • the wavelength ⁇ R of the reference light is not a wavelength that excites the fluorescent dye that is bound to the biomolecule of the irradiated object 1.
  • the wavelength lambda R of the reference light different than either of ⁇ 2' wavelengths ⁇ 1' and second fluorescence wavelength of the first fluorescent.
  • the wavelength ⁇ R of the reference light is preferably a wavelength between ⁇ 1 ′ of the first fluorescence wavelength and ⁇ 2 ′ of the second fluorescence wavelength. It is preferable that the wavelength ⁇ R of the reference light is close to both the first fluorescence and the second fluorescence, and does not overlap so as not to affect the first fluorescence and the second fluorescence.
  • Stage 4 supports support member 60.
  • the stage 4 is movable while supporting the support member 60.
  • the stage 4 is movable in each of the X-axis direction, the Y-axis direction, and the Z-axis direction while supporting the support member 60.
  • the support member 60 is supported by the stage 4 so that the surface of the irradiated object 1 (the surface on which the biomolecule is fixed) faces the first objective lens 16 of the optical system 3.
  • the stage 4 and the control device 30 are connected by a control line 51.
  • the control device 30 can move the stage 4 that supports the support member 60 in each of the X-axis direction, the Y-axis direction, and the Z-axis direction.
  • the measurement apparatus 10 replaces the next support member 60 and sequentially performs measurement.
  • the stage 4 may be configured by a stage that can rotate in the ⁇ X, ⁇ Y, and ⁇ Z directions while supporting the support member 60.
  • the optical system 3 includes an irradiation optical system 6, an imaging optical system 7, and a microlens array (MLA) 8.
  • the irradiation optical system 6 includes components for irradiating the irradiated body 1 with light emitted from the light source device 2.
  • the irradiation optical system 6 includes a first lens 11, a brightness stop (AS) 12, a field stop (FS) 13, a second lens 14, a filter block 15, and a first objective lens. 16.
  • the light emitted from the light source device 2 passes through the first lens 11, the brightness stop 12, the field stop 13, and the second lens 14 and enters the filter block 15.
  • FIG. 3 is a schematic diagram showing an example of a filter block.
  • the filter block 15 includes a first filter (first wavelength selection unit) 17 on which light from the light source device 2 enters, a dichroic mirror 18 on which light through the first filter 17 enters, and light from the dichroic mirror 18 An incident second filter (second wavelength selection unit) 19 is provided.
  • the filter block 15 is a fluorescent filter block in which an excitation filter, a dichroic mirror, and an absorption filter are integrally formed.
  • the fluorescent filter block may be called a fluorescent cube, a fluorescent mirror unit, or a fluorescent filter set.
  • the irradiation optical system 6 may include a second filter block (not shown) different from the filter block 15.
  • the second filter block is a fluorescent filter block in which an excitation filter, a dichroic mirror, and an absorption filter are integrally formed.
  • the second filter block is used when the light source device 2 emits the third excitation light (light in the third wavelength band) and the fourth excitation light (light in the fourth wavelength band). it can.
  • the dichroic mirror of the second filter block has a spectral sensitivity characteristics similar to the filter block 15 to at least the reference light lambda R.
  • a dichroic mirror of the second filter block has a predetermined transmittance (e.g.
  • the filter block 15 and the second filter block are switched by a switching unit such as a turret. Using this switching unit, one of the filter block 15 and the second filter block is disposed at a position where the light emitted from the light source device 2 enters (an optical path between the light source device 2 and the first objective lens 16). be able to.
  • the first filter 17 is a wavelength selection optical element.
  • the first filter 17 cuts a part of the wavelength region of the light from the light source device 2 and extracts the first excitation light, the second excitation light, and the reference light. It is an element.
  • the first filter 17, the wavelength band including the wavelength .lambda.1 has a wavelength band including a wavelength .lambda.2, and optical properties and the wavelength band of 100 percent transmittance of 75% including a wavelength lambda R.
  • Light in a predetermined wavelength region (first excitation light, second excitation light, and reference light) that has passed through the first filter 17 is incident on a dichroic mirror 18 that is an optical element.
  • the dichroic mirror 18 is a separation optical element that separates excitation light and fluorescence.
  • the dichroic mirror 18 is a mirror that reflects the excitation light selected by the first filter 17 and transmits the fluorescence emitted from the irradiated object 1.
  • the dichroic mirror 18 is disposed, for example, inclined by 45 degrees with respect to the optical axis.
  • the dichroic mirror 18 reflects excitation light, transmits fluorescence, partially reflects reference light, and partially transmits reflected light.
  • the dichroic mirror 18 reflects the first excitation light and the second excitation light, transmits the first fluorescence and the second fluorescence, partially reflects the reference light, and partially transmits the reflected light.
  • the dichroic mirror 18 reflects light in a wavelength band including the wavelength ⁇ 1 of the first excitation light and light in a wavelength band including the wavelength ⁇ 2 of the second excitation light, and changes the wavelength ⁇ 1 ′ of the first fluorescence.
  • the first excitation light, the second excitation light, and the reference light are reflected by the dichroic mirror 18 and guided to the first objective lens 16.
  • the first objective lens 16 is an infinite objective lens and can face the surface of the irradiated object 1 supported by the stage 4. In the present embodiment, the first objective lens 16 is disposed on the + Z side (upward) of the irradiated object 1. The first excitation light, the second excitation light, and the reference light are guided to the irradiation object 1 through the first objective lens 16. The irradiated object 1 is illuminated by the first excitation light, the second excitation light, and the reference light.
  • the second filter 19 is a wavelength selection optical element.
  • the second filter 19 selectively transmits the first fluorescence, the second fluorescence, and the reflected light.
  • the second filter 19 is an absorption filter.
  • the absorption filter may be called an emission filter or a barrier filter.
  • the imaging optical system 7 forms an image of the light beam (first fluorescence, second fluorescence, and reflected light) from the irradiated object 1 in the vicinity of its focal plane (imaging plane).
  • the imaging optical system 7 includes a second objective lens 21.
  • the microlens array 8 and the sensor 5 are arranged in that order near the focal plane of the second objective lens 21.
  • the microlens array 8 includes a plurality of microlenses arranged two-dimensionally. As an example, the microlens array 8 is disposed on the imaging plane of the second objective lens 21. Note that the microlens array 8 may be disposed on the pupil plane of the second objective lens 21. The number of lenses in the vertical direction and the number of lenses in the horizontal direction (arrangement density) of the microlens array 8 are appropriately set according to the resolution required for the image acquired by the measurement apparatus 10.
  • the sensor 5 is a light receiving unit in which light receiving elements (photodiodes) are arranged two-dimensionally.
  • the sensor 5 is an image sensor.
  • the image pickup device includes a charge coupled device (CCD) and a complementary metal oxide semiconductor (CMOS).
  • CMOS complementary metal oxide semiconductor
  • the sensor 5 may be called a photo sensor array, an image sensor, or an area sensor.
  • the sensor 5 receives light from the irradiated object 1 via the microlens array 8.
  • the sensor 5 outputs a signal corresponding to the received light amount to the control device 30.
  • FIG. 4 is a schematic diagram showing the arrangement of the microlens array 8 and the sensor 5 in which the light receiving elements are two-dimensionally arranged.
  • the sensor 5 includes a pixel array that receives light that has passed through each microlens 81 (that is, an arrangement pattern corresponding to the microlens 81).
  • a signal obtained from the light receiving element of the sensor 5 that receives light that has passed through the microlens 81 is referred to as a “pixel signal”.
  • the control device 30 performs predetermined signal processing on the set of pixel signals acquired by the sensor 5. The contents of the signal processing will be described later.
  • the control device 30 displays the signal processing result using the display device 40.
  • the display device 40 can display the image information of the irradiated object 1 acquired by the sensor 5.
  • a set of pixel signals at a plurality of different focal positions can be obtained without performing focusing by the AF function for each fluorescence.
  • the control device 30 can perform predetermined signal processing on a set of pixel signals acquired by the sensor 5 to generate a plurality of images having different focal positions.
  • FIG. 5 is an example in which a plurality of images having different focal positions are acquired using the microlens array 8 and the sensor 5. The vertical axis in FIG.
  • the control device 30 can extract a signal sequence of pixel signals having a common (that is, the same) focal length, and generate an image at the focal length.
  • FIG. 5 shows a plurality of images generated from pixel signals at the respective focal lengths Z 1 to Z 6 .
  • FIGS. 6 and 7 are diagrams for explaining pixel signal extraction processing in a configuration using the microlens array 8 and the sensor 5 in which the light receiving elements are two-dimensionally arranged.
  • each light ray incident on five pixels (a, b, c, d, e) arranged in a straight line in the sensor 5 (a principal ray passing through the center of the corresponding microlens). Only) is shown. Further, each element in each drawing is given a suffix (1, 2, 3,%) For indicating coordinates in a plane perpendicular to the optical axis.
  • L (4) (a4 + b4 + c4 + d4 + e4) It becomes.
  • Y-axis direction in addition to the X-axis direction.
  • pixel signals may be extracted in the same manner in the Y-axis direction.
  • the image signal of the coordinate X′4 in the center area can be obtained from the following equation (2).
  • FIG. 8 is a diagram for explaining the amount of chromatic aberration of each fluorescence.
  • the wavelength of the first fluorescence (first color light) is ⁇ 1 ′
  • the wavelength of the second fluorescence (second color light) is ⁇ 2 ′.
  • the amount of chromatic aberration of the first fluorescence is different from the amount of chromatic aberration of the second fluorescence.
  • the focused position of the image of the reflected light of the wavelength lambda R and Z 0. In this case, the focal point of the first fluorescence is shifted from Z 0 by ⁇ Z 1 .
  • the focal point of the second fluorescence is shifted from Z 0 by ⁇ Z 2 .
  • the following processing can be executed after acquiring a set of pixel signals at a plurality of different focal positions using the microlens array 8 and the sensor 5.
  • the control device 30 extracts the pixel signal from the light receiving element on which the light at the focal position Z 0 + ⁇ Z 1 is incident, from a set of pixel signals at a plurality of different focal positions.
  • the signal sequence of the pixel signals extracted here substantially corresponds to a signal sequence that represents a fluorescence image at the focal point of the first fluorescence.
  • the control device 30 extracts a pixel signal from the light receiving element on which the light at the focal position Z 0 + ⁇ Z 2 is incident, from a set of pixel signals at a plurality of different focal positions.
  • the signal sequence of the pixel signals extracted here substantially corresponds to a signal sequence that represents a fluorescence image at the focal point of the second fluorescence.
  • the imaging position of the reflected light (focus point) Z 0 as a reference explain the advantages of obtaining the focus and the focal point of the second fluorescence of the first fluorescent.
  • the fluorescent image may be dark or low in contrast depending on the target specimen. In such a situation, it may be difficult to obtain the signal sequence of the first fluorescent focused pixel signal and the signal sequence of the second fluorescent focused pixel signal. Therefore, it is preferable to obtain Z 0 from an image of reflected light from which sufficient contrast is obtained, and use Z 0 as a reference.
  • FIG. 9 is a diagram for explaining the components of the control device 30 that realizes the above-described processing.
  • the control device 30 includes a signal sequence extraction unit 31, a signal sequence processing unit 32, a stage control unit 33, and a chromatic aberration information storage unit 34.
  • the processing of the signal sequence extraction unit 31, the signal sequence processing unit 32, and the stage control unit 33 can be realized by a program code of software that realizes these functions.
  • the processor of the control device 30 executes processing described below in accordance with an instruction of a predetermined program stored in the memory. Note that some processes of the signal sequence extraction unit 31, the signal sequence processing unit 32, and the stage control unit 33 may be realized by hardware using electronic components such as an integrated circuit.
  • the chromatic aberration information storage unit 34 may be realized by a storage device of the control device 30.
  • the signal string extraction unit 31 receives pixel signals from each element of the sensor 5 that receives light that has passed through the microlens array 8.
  • Signal sequence extraction unit 31 from a set of pixel signals of the reflected light at a plurality of different focus, determine the reference focus position Z 0 of the reflected light.
  • Z 0 as the focal point position of the image of the reflected light
  • reference focus position Z 0 here, for example, contrast the focal position where the maximum, not exactly coincide with the focus position In some cases.
  • the reference focal position obtained here is regarded as the in-focus position. If not exactly match the focused position, as an example, the position closest to the maximum value of the contrast and Z 0.
  • the focal position where the contrast is maximized is almost the focal position.
  • the signal sequence extraction unit 31 extracts a signal sequence of pixel signals corresponding to the focal point of the first fluorescence from a set of first pixel signals at a plurality of different focal points.
  • the first set of pixel signals is a set of pixel signals acquired during a time period in which the first fluorescence will be emitted.
  • the expression “first set of pixel signals” means the same contents.
  • the signal sequence extraction unit 31 extracts the signal sequence of the pixel signal corresponding to the focal point of the first fluorescence using the chromatic aberration amount ⁇ Z 1 of the first fluorescence from the reference focal position Z 0 .
  • the signal sequence of the first fluorescence pixel signal extracted here substantially corresponds to a signal sequence representing a fluorescence image at the focal point of the first fluorescence.
  • the signal sequence extraction unit 31 extracts a signal sequence of pixel signals corresponding to the focal point of the second fluorescence from a set of second pixel signals at a plurality of different focal points.
  • the second set of pixel signals is a set of pixel signals acquired during a time period in which the second fluorescence will be emitted.
  • the expression “second set of pixel signals” means the same content.
  • the signal string extracting unit 31 using the chromatic aberration amount [Delta] Z 2 of the second fluorescence from the reference focus position Z 0, and extracts the signal sequence of the pixel signals corresponding to the focal point of the second fluorescence.
  • the signal sequence of the second fluorescence pixel signal extracted here substantially corresponds to a signal sequence representing a fluorescence image at the focal point of the second fluorescence.
  • the signal sequence processing unit 32 performs the first signal processing on the signal sequence of the pixel signal corresponding to the focal point of the first fluorescence, and the signal sequence of the pixel signal corresponding to the focal point of the second fluorescence.
  • the second signal processing is executed.
  • the first signal processing is processing for generating a first image from a signal sequence of pixel signals corresponding to the focal point of the first fluorescence.
  • the second signal processing is processing for generating a second image from a signal sequence of pixel signals corresponding to the focal point of the second fluorescence.
  • the control device 30 may output the first image and the second image to the display device 40.
  • the first signal processing is a process of outputting information representing the signal value of the first fluorescence pixel signal and position information of the irradiated object 1 corresponding to the first fluorescence pixel signal.
  • the information indicating the signal value of the pixel signal is a luminance value.
  • the position information of the irradiated object 1 is a spot address.
  • the signal sequence processing unit 32 extracts a pixel signal having a luminance value exceeding a predetermined threshold value. When the luminance value exceeds a predetermined threshold value, it can be considered that the first fluorescence is generated.
  • the signal sequence processing unit 32 associates the luminance value with the address of the spot corresponding to the luminance value in the irradiated object 1.
  • the signal sequence processing unit 32 may output the associated result to the display device 40. Thereby, it can be determined in which position the irradiated body 1 emits fluorescence.
  • the second signal processing may be processing for outputting information representing the signal value of the second fluorescence pixel signal and position information of the irradiated object 1 corresponding to the second fluorescence pixel signal. .
  • the stage control unit 33 determines whether the signal value of any pixel signal in the set of reflected pixel signals is greater than a predetermined threshold value.
  • the signal value used for determination is a luminance value.
  • the threshold value may be preset based on the depth of focus or the noise level.
  • the stage control unit 33 may calculate contrast from a set of pixel signals of reflected light and determine whether the contrast value is greater than a predetermined threshold value. Note that the above determination may be performed using the pixel signals of the first pixel signal set and the second pixel signal set.
  • One of the advantages of the measurement apparatus 10 described here is that when performing measurement on the same irradiated object 1, when the pixel signal is acquired by irradiating the first excitation light, the second excitation light is obtained. It is not necessary to drive the optical system of the measuring apparatus 10 (for example, a lens for focusing (focusing lens)) for focusing between the time when the pixel signal is acquired by irradiating the lens.
  • One of the advantages of the measuring apparatus 10 is that when performing measurement on the same irradiated object 1, a pixel signal is obtained by irradiating the first excitation light, and a pixel by irradiating the second excitation light. It is not necessary to drive the stage 4 for focusing between the time when the signal is acquired.
  • the height of the stage 4 is adjusted once (in this case, it is not always necessary to focus), and when the pixel signal is acquired by irradiating the first excitation light There is no need to drive the lens or stage 4 of the optical system for focus adjustment between the time when the pixel signal is acquired by irradiating the second excitation light.
  • the stage control unit 33 extracts an arbitrary pixel signal from a set of pixel signals of reflected light, first fluorescence, and second fluorescence, and a predetermined threshold value is included in these pixel signals. It may be determined whether there is a signal having a larger luminance value.
  • the stage control unit 33 may change or adjust the height direction (Z direction) of the stage 4 when there is no signal having a luminance value larger than a predetermined threshold value.
  • the stage control unit 33 extracts a predetermined number of pixel signals from the set of reflected pixel signals, and determines whether or not a luminance value larger than a predetermined threshold exists in the pixel signals. You may judge. By preferentially handling a set of pixel signals of reflected light that is highly likely to obtain sufficient contrast, it is possible to determine whether the focal position is greatly deviated.
  • the chromatic aberration information storage unit 34 includes information on the amount of chromatic aberration of each fluorescence (hereinafter referred to as “chromatic aberration information”). Accordingly, it is possible to acquire a signal sequence at the focal point position of each fluorescence in accordance with the amount of chromatic aberration of each fluorescence.
  • the chromatic aberration information includes at least a chromatic aberration amount ⁇ Z 1 of the first fluorescence from the focal position of the reflected light and a chromatic aberration amount ⁇ Z 2 of the second fluorescence from the focal position of the reflected light.
  • the chromatic aberration information is information in which information on the wavelength of the excitation light is associated with information on the amount of chromatic aberration of fluorescence generated from the excitation light.
  • the control device 30 controls the light source device 2 to emit arbitrary excitation light, the control device 30 can acquire information on the amount of chromatic aberration of fluorescence corresponding to the excitation light.
  • the chromatic aberration information may include information indicating the relationship of the chromatic aberration amount between the first fluorescence and the second fluorescence. Using this relationship, the control device 30 may obtain the relationship between the in-focus position of the first fluorescence and the in-focus position of the second fluorescence. Thus, if one of the in-focus position of the first fluorescence and the in-focus position of the second fluorescence is known without obtaining the reference point position of the reflected light, the other in-focus position can be obtained.
  • the chromatic aberration information in the chromatic aberration information storage unit 34 can be set in advance based on the design of the optical system (lens or the like) 3.
  • the chromatic aberration amount of each fluorescence may be measured at the time of the first measurement of the measuring apparatus 10 and the measurement result may be stored in the chromatic aberration information storage unit 34 as chromatic aberration information.
  • FIG. 10 is an example of a flowchart when measuring the irradiated object 1 on the support member 60 by the measuring apparatus 10.
  • the support member 60 is prepared (1001).
  • a support member 60 for example, a plate
  • a set of pixel signals at multiple focal points is acquired (1002).
  • signal processing is performed on the set of pixel signals at multiple focal points (1003).
  • FIG. 11 is an example of detailed contents of the flowchart of FIG. In the following, the processing in which the control device 30 and each component of the control device 30 are the subject may be described with the processor as the subject.
  • the irradiated object 1 (for example, one irradiated object among a plurality of irradiated objects 1 supported by the support member 60) is the first excitation light, the second excitation light, and the reference light. It is irradiated with three lights.
  • the light source device 2 emits first excitation light, reference light, and second excitation light in the order of first excitation light, reference light, and second excitation light.
  • the period in which the light source device 2 emits the first excitation light, the period in which the light source device 2 emits the reference light, and the period in which the light source device 2 emits the second excitation light do not overlap. .
  • the irradiated object 1 is irradiated in the order of the first excitation light, the reference light, and the second excitation light.
  • the period in which the light source device 2 emits the reference light is between the period in which the light source device 2 emits the first excitation light and the period in which the light source device 2 emits the second excitation light.
  • the reason why the reference light is irradiated in this manner between the first excitation light and the second excitation light is as follows. It is conceivable that the fluorescence information acquisition time (the charge accumulation time of the sensor 5 with respect to the fluorescence emitted from the irradiated object 1) is longer than the information acquisition time of the reflected light (the charge accumulation time of the sensor 5 with respect to the reflected light).
  • the second fluorescence information is obtained after obtaining the reflected light information (a set of pixel signals).
  • the time to get is longer.
  • Z direction deviation causes a deviation of the reference focus position Z 0 as a reference.
  • the irradiation order of the reference beam is preferably close to each of the first excitation light and second excitation light.
  • the emission order of the first excitation light, the second excitation light, and the reference light is not limited to this, and may be any order.
  • Step 1002 in FIG. 10 includes steps 1101 to 1108.
  • Step 1003 in FIG. 10 includes steps 1109 to 1112.
  • the support member 60 is prepared (1001). As an example, the support member 60 is disposed on the stage 4.
  • the light source device 2 irradiates the irradiated body 1 with the first excitation light based on the signal from the control device 30 (1101).
  • the signal sequence extraction unit 31 acquires a set of first pixel signals at a plurality of different focal points from the sensor 5 (1102).
  • a set of pixel signals during a time period in which the first fluorescence is emitted from the irradiated body 1 by irradiating the irradiated body 1 with the first excitation light is acquired.
  • the set of first pixel signals acquired here may or may not include the first fluorescence information.
  • each light receiving element of the sensor 5 accumulates charge between when the charge accumulation start instruction is given and when the charge accumulation end instruction is given (that is, the sensor according to the charge accumulation start instruction 5 each of the light receiving elements 5 starts charge accumulation, and each light receiving element of the sensor 5 finishes charge accumulation in response to a charge accumulation end instruction), and the sensor 5 that has performed the accumulation operation (after the accumulation operation)
  • the light source device 2 irradiates the irradiated object 1 with reference light based on a signal from the control device 30 (1103).
  • the irradiated object 1 irradiated with the reference light is the same as the irradiated object 1 irradiated with the first excitation light in Step 1101.
  • the signal string extraction unit 31 of the control device 30 acquires a set of pixel signals of reflected light at a plurality of different focal points from the sensor 5 (1104).
  • a set of pixel signals of reflected light is stored in each light receiving element of the sensor 5 from when the charge accumulation start instruction is issued until the charge accumulation end instruction is issued (that is, the sensor is activated according to the charge accumulation start instruction).
  • each of the light receiving elements 5 starts charge accumulation, and each light receiving element of the sensor 5 finishes charge accumulation in response to a charge accumulation end instruction), and the sensor 5 that has performed the accumulation operation (after the accumulation operation)
  • the imaging operation for obtaining the first set of pixel signals and the imaging operation for obtaining the set of reflected light pixel signals are performed individually. The period during which the imaging operation for obtaining the set of reflected pixel signals does not overlap with the period during which the imaging operation for obtaining the first set of pixel signals is performed.
  • the light source device 2 irradiates the irradiated body 1 with the second excitation light based on the signal from the control device 30 (1105).
  • the signal string extraction unit 31 acquires a set of second pixel signals at a plurality of different focal points from the sensor 5 (1106).
  • a set of pixel signals during a time period in which the second fluorescence is emitted after irradiation of the second excitation light to the irradiated object 1 is acquired.
  • the set of second pixel signals acquired here may or may not include the second fluorescence information.
  • each light receiving element of the sensor 5 accumulates charge between when the charge accumulation start instruction is given and when the charge accumulation end instruction is given (that is, the sensor according to the charge accumulation start instruction).
  • each of the light receiving elements 5 starts charge accumulation, and each light receiving element of the sensor 5 finishes charge accumulation in response to a charge accumulation end instruction), and the sensor 5 that has performed the accumulation operation (after the accumulation operation)
  • This is a set of pixel signals obtained by performing an imaging operation including a reading operation of reading (outputting) charges from each light receiving element once.
  • An imaging operation for obtaining a first set of pixel signals, an imaging operation for obtaining a set of reflected pixel signals, and an imaging operation for obtaining a second pixel signal are performed separately.
  • the period during which the imaging operation for obtaining the second set of pixel signals is performed is the period during which the imaging operation for obtaining the first set of pixel signals is performed and the imaging operation for obtaining the set of reflected pixel signals. Does not overlap with any period during which
  • the stage control unit 33 extracts an arbitrary pixel signal from the set of reflected pixel signals, and determines whether any of the pixel signals has a signal value larger than a predetermined threshold value ( 1107).
  • the signal value is a luminance value.
  • the threshold value may be preset based on the depth of focus or the noise level.
  • the stage control unit 33 may calculate contrast from a set of pixel signals of reflected light and determine whether the contrast value is greater than a predetermined threshold value.
  • the stage control unit 33 extracts an arbitrary pixel signal from a set of pixel signals of the first fluorescence or the second fluorescence instead of the reflected light, and a predetermined threshold value is included in these pixel signals.
  • the luminance value of the first fluorescence or the second fluorescence can be compared with a predetermined threshold value.
  • the process proceeds to step 1109. If the condition is not satisfied, it is determined that the focal position is significantly shifted, and the process proceeds to step 1108.
  • the stage control unit 33 drives the stage 4 (1108).
  • the stage control unit 33 changes or adjusts the height (Z direction) of the stage 4.
  • the stage control unit 33 changes or adjusts the height of the stage 4 by a predetermined height.
  • the stage control unit 33 changes or adjusts the amount of chromatic aberration [Delta] Z 1 or chromatic aberration [Delta] Z height of only 2 minutes Stage 4 of the second fluorescence of the first fluorescent. After changing or adjusting the height of the stage 4, the process returns to Step 1101.
  • Signal string extraction unit 31 determines a set of pixel signals of the reflected light having a set of a plurality of different focus each pixel signal, a reference focus position Z 0 (1109).
  • Figure 12 is an example of a method for determining the reference focus position Z 0.
  • the signal sequence extraction unit 31 generates an image from a signal sequence of pixel signals at a common focal position, and holds a plurality of images at a plurality of focal positions.
  • the signal sequence extraction unit 31 performs the following processing using information of a plurality of images. Note that it is not always necessary to treat the image as an image, and the processing described below may be performed as a set of pixel signals.
  • the signal sequence extraction unit 31 divides an image of each focal position Z ⁇ 3 to Z 3 into a plurality of regions. As an example, the signal sequence extraction unit 31 calculates the contrast of a region corresponding to the same position in the images at the respective focal positions Z ⁇ 3 to Z 3 . As an example, the signal sequence extraction unit calculates the contrast of the central region of the images at the focal positions Z ⁇ 3 to Z 3 (shaded portion in FIG. 12). The area for calculating the contrast may be another area or all areas on the image.
  • FIG. 13 is an example of the result of calculating the contrast from the set of pixel signals of reflected light, and shows the contrast of the central region of the image at each focal position Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 determines a position where the contrast is maximum from the focal positions Z ⁇ 3 to Z 3 as the reference focal position Z 0 .
  • the maximum value of the approximate curve in FIG. 13 is located between two Z positions, the closer one may be determined as the reference focal position Z 0 .
  • the signal string extraction unit 31 acquires information on the first fluorescence chromatic aberration amount ⁇ Z 1 from the chromatic aberration information storage unit 34.
  • the signal sequence extraction unit 31 extracts a signal sequence of a pixel signal corresponding to the focal point of the first fluorescence using the chromatic aberration amount ⁇ Z 1 of the first fluorescence from the reference focal position Z 0 (1110).
  • FIG. 14 is an example of acquiring a signal sequence of pixel signals corresponding to the focal point of the first fluorescence from a set of first pixel signals at a plurality of different focal points Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 generates an image from a signal sequence of pixel signals at a common focal position, and holds a plurality of images at a plurality of focal positions.
  • the signal sequence extraction unit 31 extracts an image at the in-focus position of the first fluorescence from the images at the plurality of focus positions Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 uses the first fluorescence chromatic aberration amount ⁇ Z 1 from the reference focal position Z 0 to extract an image of the focal position Z 2 corresponding to the focal point of the first fluorescence. It is not necessarily treated as an image, signal sequence extraction unit 31, from the set of pixel signals may extract a signal sequence corresponding to the focus position Z 2.
  • the signal sequence extraction unit 31 acquires information on the second fluorescence chromatic aberration amount ⁇ Z 2 from the chromatic aberration information storage unit 34.
  • Signal string extracting unit 31 using the chromatic aberration amount [Delta] Z 2 of the second fluorescence from the reference focus position Z 0, and extracts the signal sequence of the pixel signals corresponding to the focal point of the second fluorescent (1111).
  • FIG. 15 is an example of acquiring a signal sequence of pixel signals corresponding to the focal point of the second fluorescence from a set of second pixel signals at a plurality of different focal points Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 generates an image from a signal sequence of pixel signals at a common focal position, and holds a plurality of images at a plurality of focal positions.
  • the signal sequence extraction unit 31 extracts an image at the focal point position of the second fluorescence from the images of the plurality of focal positions Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 uses the second fluorescence chromatic aberration amount ⁇ Z 2 from the reference focal position Z 0 to extract an image at the focal position Z ⁇ 2 corresponding to the focal point of the first fluorescence. .
  • the signal sequence extraction unit 31 may extract the signal sequence corresponding to the focal position Z- 2 from the set of pixel signals.
  • the reference focal position Z 0 + the chromatic aberration amount ⁇ Z 1 or ⁇ Z 2 may not coincide with a plurality of different focal points Z ⁇ 3 to Z 3 .
  • Z 0 + ⁇ Z 1 was Z 2.4 .
  • Z 2.4 is close found the following Z 2 as compared to Z 3, may extract a signal string corresponding to the closer Z 2.
  • the signal sequence extraction unit 31 may generate a signal value corresponding to Z 2.4 by multiplying each of the signal value of the Z 2 pixel signal and the signal value of the Z 3 pixel signal by a weighting factor. .
  • a weighted average may be used. Weighting factor applied to the signal value of the pixel signals of the signal value and Z 3 of the pixel signals of Z 2 may be set to the same, it is set differently according to the distance from the reference focus position Z 0 Good.
  • the focal point position of the reflected light is between the focal point position of the first fluorescence and the second focal point position.
  • the relationship between the fluorescence and the second fluorescence is not limited to this example.
  • the focal point position of the reflected light may be on the positive direction side (+ Z side) as compared with the first fluorescence.
  • the focal position of the reflected light may be on the negative direction side (+ Z side) as compared with the second fluorescence.
  • the chromatic aberration information in the chromatic aberration information storage unit 34 only needs to include information for specifying the relationship between the reflected light, the first fluorescence, and the second fluorescence.
  • the signal sequence processing unit 32 performs first signal processing on the signal sequence of the pixel signal corresponding to the extracted first fluorescence focal point, and extracts the extracted second fluorescence focal point.
  • Second signal processing is executed for the signal sequence of pixel signals corresponding to (1112).
  • the signal sequence processing unit 32 generates a first result from the signal sequence of the pixel signal corresponding to the extracted focal point of the first fluorescence, and the pixel sequence of the pixel signal corresponding to the extracted focal point of the second fluorescence.
  • a second result is generated from the signal sequence.
  • the first result is an image (see FIG. 14) corresponding to the focal point of the first fluorescence.
  • the second result is an image (see FIG. 15) corresponding to the focal point of the second fluorescence.
  • the signal sequence processing unit 32 uses the extracted image corresponding to the focal point of the first fluorescence (see FIG. 14). The image may be output to the display device 40 as it is, and the extracted image (see FIG. 15) corresponding to the focal point of the second fluorescence may be output to the display device 40 as it is.
  • the signal sequence processing unit 32 performs the first processing from the extracted signal sequence of the pixel signals corresponding to the focal point of the first fluorescence.
  • An image may be generated, and a second image may be generated from a signal sequence of pixel signals corresponding to the extracted focal point of the second fluorescence.
  • the first result includes information indicating the signal value of the first fluorescence pixel signal with respect to the signal sequence of the pixel signal corresponding to the focal point of the extracted first fluorescence, and the first result. And position information of the irradiated object 1 corresponding to the fluorescent pixel signal.
  • the second result is that the information indicating the signal value of the second fluorescence pixel signal and the pixel signal of the second fluorescence are related to the signal sequence of the pixel signal corresponding to the focal point of the extracted second fluorescence. Corresponding position information of the irradiated object 1.
  • the information representing the signal value is a luminance value.
  • the position information of the irradiated object 1 is a spot address. It can be said that the 1st result and the 2nd result have shown the result of having made the to-be-irradiated body 1 and the sample react. It can be said that the assayed result includes the first result and the second result.
  • the first result and the second result are stored in the storage device.
  • the storage device may be a storage device inside the measurement device 10 (for example, inside the control device 30) or a storage device (external device, external device) outside the measurement device 10. Examples of the storage device outside the measuring apparatus 10 include a server, a printer, and a portable information terminal.
  • the first result and the second result generated by the signal sequence processing unit 32 are output to the external device by the control device 30.
  • Communication between the measuring apparatus 10 and the external device may be wired communication or wireless communication.
  • the first result and the second result output by the control device 30 are stored in the external device.
  • the external device is a printer
  • the first result and the second result may be printed on paper (for example, automatically) and output.
  • the external device is a portable information terminal
  • the first result and the second result may be displayed on the display unit of the portable information terminal (for example, automatically).
  • the reference light irradiation period may be set before and after the first excitation light irradiation period, and the reference light irradiation period may be set before and after the second excitation light irradiation period.
  • steps 1103 and 1104 may be added before step 1101, and steps 1103 and 1104 may be added after step 1106.
  • a set of signals (first set) at multiple focal points of the reflected light is acquired before the irradiation of the first excitation light, and then after the irradiation of the first excitation light and the second Before the excitation light irradiation, a set of signals (second set) at the multifocal point of the reflected light is obtained, and after the second excitation light irradiation, the pixel signals of the reflected light at the plurality of focus positions are obtained.
  • a set (third set) is acquired.
  • the signal sequence extraction unit 31 obtains the reference focal position Z 0A obtained from the first set, the reference focal position Z 0B obtained from the second set, and the reference focal position Z 0C obtained from the third set. May be compared. As an example, the signal sequence extraction unit 31 may determine whether the reference focal positions Z 0A to Z 0C are all the same value. When the reference focal positions Z 0A to Z 0C are different, the signal sequence extraction unit 31 may output which of the reference focal positions Z 0A to Z 0C is different. The user can grasp at which point the position shift in the Z direction has occurred.
  • ⁇ Z 1 ⁇ Z 2 .
  • FIG. 16 and FIG. 17 are used to explain another flowchart for measuring the irradiated object 1.
  • Step 1002 in FIGS. 16 and 17 is composed of steps 1101 to 1108 in FIG.
  • Step 1003 in FIGS. 16 and 17 is composed of steps 1109 to 1112 in FIG.
  • a support member 60 for example, a plate
  • a set of signals (first set) of the object 1 to be measured is acquired (1002).
  • movement to the next irradiated body 1 (1004) and acquisition of a set of signals of the next irradiated body 1 (second set) (1002) is performed.
  • the operation of moving the measurement object to the next irradiated object 1 is performed by driving the stage 4.
  • the movement to the next irradiated object 1 is performed after the signal processing (1003).
  • FIG. 11 the movement to the next irradiated object 1 is performed after the signal processing (1003).
  • the signal processing (1003) and the movement to the next irradiated object 1 (1004) are performed.
  • acquisition (1002) of the next set of signals (second set) of the irradiated object 1 are executed in parallel. Thereby, the throughput of image acquisition is improved, and more information on the irradiated object 1 can be acquired in a short time.
  • a support member 60 for example, a plate
  • a set of signals of a certain irradiated body 1 is acquired (1002), and then moved to the next irradiated body 1 (1004).
  • the operation of moving the measurement object to the next irradiated object 1 is performed by driving the stage 4.
  • the combination of step 1002 and step 1004 is repeated up to the last irradiated object 1 on the support member 60.
  • signal processing is collectively performed on the set of signals of all the irradiated objects 1 (1003). In this flow, every time a set of signals is acquired (1002), signal processing (1003) is not performed, and information of all the irradiated objects 1 is acquired in advance.
  • step 1003 is advantageous when step 1003 is performed by another control device.
  • Information on all the irradiated objects 1 acquired in advance is transferred to another control device via a recording medium or a network.
  • Another control device may collectively perform signal processing (1003) on a set of signals of all the irradiated objects 1.
  • the signal sequence corresponding to the focal point of the first fluorescence corresponding to the first excitation light and the second excitation light can be handled without focusing using the AF function.
  • a signal sequence corresponding to the focal point of the second fluorescence can be obtained.
  • two fluorescences having different wavelengths have different amounts of chromatic aberration, so that a focusing process using an AF function is required every time an image is acquired.
  • an operation such as driving the lens of the optical system or changing the height of the stage is required each time an image of each fluorescence is acquired by the AF function. Therefore, the time required for driving the lens and changing the height of the stage has had a great influence on the throughput of image acquisition.
  • the AF function may cause an error.
  • the AF function is used between when the pixel signal is acquired by irradiating the first excitation light and when the pixel signal is acquired by irradiating the second excitation light. It does not have to be focused.
  • an optical system lens for focusing between when the pixel signal is acquired by irradiating the first excitation light and when the pixel signal is acquired by irradiating the second excitation light. Processing such as driving the focusing lens or changing the height of the stage is not necessary. In the example of FIG.
  • steps 1101 to 1106 an operation such as driving a lens (focusing lens) of the optical system or changing the height of the stage is not necessary.
  • focusing by the AF function is unnecessary during steps 1101 to 1106, and a signal sequence corresponding to the focal point of the first fluorescence from the set of acquired pixel signals at a plurality of focal positions, and the first A signal sequence corresponding to the focal point of the two fluorescences can be extracted. Therefore, the throughput of image acquisition is greatly improved, and more images of the irradiated object 1 can be acquired in a short time.
  • FIG. 18 is an example of a flowchart for measuring the irradiated object 1 by the measuring apparatus 10.
  • the light source device 2 emits reference light, first excitation light, and second excitation light simultaneously.
  • the support member 60 is placed on the stage 4 (1801).
  • the light source device 2 emits both the reference light, the first excitation light, and the second excitation light based on the signal from the control device 30 (1802).
  • the reference light irradiation period, the first excitation light irradiation period, and the second excitation light irradiation period may overlap.
  • the reference light irradiation period, the first excitation light irradiation period, and the second excitation light irradiation period may at least partially overlap each other.
  • irradiation with reference light, irradiation with first excitation light, and irradiation with second excitation light are started simultaneously.
  • the signal string extraction unit 31 of the control device 30 acquires a set of pixel signals at a plurality of different focal points from the sensor 5 (1803).
  • the set of pixel signals at a plurality of different focal points here includes a set of reflected light pixel signals, a set of first pixel signals, and a set of second pixel signals.
  • the set of pixel signals at a plurality of different focal points is one. That is, by performing one imaging operation on the same subject 1, a set of reflected pixel signals, a set of first pixel signals, and a set of second pixel signals are obtained.
  • the reference light, the first excitation light, and the second excitation light are irradiated at different times, three sets of pixel signals at a plurality of different focal points are acquired.
  • three imaging operations are performed on the same irradiated object 1 in order to obtain a set of pixel signals of reflected light, a set of first pixel signals, and a set of second pixel signals. Do. That is, for the same subject 1, one imaging operation is performed to obtain a set of reflected light pixel signals, one imaging operation is performed to obtain a first pixel signal set, and second In order to obtain a set of pixel signals, one imaging operation is performed.
  • the stage control unit 33 extracts an arbitrary pixel signal from a set of pixel signals at a plurality of different focal points, and determines whether any of the pixel signals has a signal value larger than a predetermined threshold value. (1804). As an example, the signal value is a luminance value. If the condition is satisfied, the process proceeds to step 1806. If the condition is not satisfied, the process proceeds to step 1805. In this case, the stage control unit 33 drives the stage 4. As an example, the stage control unit 33 changes or adjusts the height (Z direction) of the stage 4 (1805). The content of step 1805 is the same as that of 1108 in FIG.
  • the signal sequence extraction unit 31 determines the reference focal position Z 0 from a set of pixel signals at a plurality of different focal points (1806). As an example, the signal sequence extraction unit 31 generates an image from a signal sequence of pixel signals at a common focal position, and holds a plurality of images at a plurality of focal positions. The signal sequence extraction unit 31 performs the following processing using information of a plurality of images. Note that it is not always necessary to treat the image as an image, and the processing described below may be performed as a set of pixel signals.
  • the signal sequence extraction unit 31 divides the image at each focal position into a plurality of regions. As an example, the signal sequence extraction unit 31 calculates the contrast of the region corresponding to the same position in the image at each focal position. As an example, the signal sequence extraction unit 31 calculates the contrast of the central region of the image at each focal position.
  • the area for calculating the contrast may be another area or all areas on the image.
  • Figure 19 is an example of a method for determining the reference focus position Z 0.
  • FIG. 19 shows the relationship between the focal position and contrast. If the reflected light, the first fluorescence, and the second fluorescence are not substantially affected by light of other wavelengths at the respective in-focus positions, a contrast graph is obtained as shown by the approximate curve in FIG. Has three peaks.
  • the signal sequence extraction unit 31 extracts three focal positions corresponding to three peaks from a graph representing the relationship between the focal position and the contrast.
  • a known method can be used for peak extraction.
  • the peak may be extracted using information of the first derivative or second derivative of the approximate curve representing the relationship between the focal position and the contrast.
  • the chromatic aberration information stored in the chromatic aberration information storage unit 34 includes the chromatic aberration amount ⁇ Z 1 of the first fluorescence from the focal position of the reflected light and the chromatic aberration of the second fluorescence from the focal position of the reflected light. and a quantity [Delta] Z 2 at least.
  • the signal sequence extraction unit 31 refers to the chromatic aberration information so that (i) the in-focus position of the first fluorescence is on the positive direction side (+ Z side) with respect to the in-focus position of the reflected light.
  • the signal string extraction unit 31 determines that the central peak among the three peaks in FIG. 19 is the focused position of the reflected light (that is, the reference focal position Z 0 ) by referring to the chromatic aberration information.
  • the signal string extraction unit 31 acquires information on the first fluorescence chromatic aberration amount ⁇ Z 1 from the chromatic aberration information storage unit 34.
  • the signal sequence extraction unit 31 extracts the signal sequence of the pixel signal corresponding to the focal point of the first fluorescence by using the chromatic aberration amount ⁇ Z 1 of the first fluorescence from the reference focal position Z 0 (1807).
  • the signal sequence extraction unit 31 acquires information on the second fluorescence chromatic aberration amount ⁇ Z 2 from the chromatic aberration information storage unit 34.
  • Signal string extracting unit 31 using the chromatic aberration amount [Delta] Z 2 of the second fluorescence from the reference focus position Z 0, and extracts the signal sequence of the pixel signals corresponding to the focal point of the second fluorescent (1808).
  • the reference focal position Z 0 + chromatic aberration amount ⁇ Z 1 or ⁇ Z 2 may not coincide with a plurality of different focal points Z ⁇ 5 to Z 5 .
  • Z 0 + ⁇ Z 1 was Z 4.4 .
  • Z 4.4 is close found the following Z 4 as compared to Z 5, may extract a signal string corresponding to the closer Z 4.
  • the signal sequence extraction unit 31, by multiplying a weighting factor to each of the signal value of the signal value of the pixel signal and the pixel signal of the Z 5 of Z4, may generate a signal value corresponding to Z 4.4.
  • a weighted average may be used.
  • Weighting factor applied to the signal value of the pixel signal of the pixel signal the signal values and Z 5 of Z 4 may be set to the same, it is set differently according to the distance from the reference focus position Z 0 Good.
  • the signal sequence processing unit 32 performs first signal processing on the signal sequence of the extracted first fluorescence pixel signal, and performs the extraction on the signal sequence of the extracted second fluorescence pixel signal.
  • the second signal processing is then executed (1809). Note that the content of step 1809 is the same as that of step 1112 in FIG.
  • the focal point of the first fluorescence corresponding to the first excitation light even when the light source device 2 emits both the reference light, the first excitation light, and the second excitation light, the focal point of the first fluorescence corresponding to the first excitation light. And a signal sequence corresponding to the focal point of the second fluorescence corresponding to the second excitation light can be obtained. For example, it may take several seconds to irradiate the excitation light and obtain sufficient information on the corresponding fluorescence.
  • the reference light, the first excitation light, and the second excitation light are emitted together, the first excitation light and the second excitation light are sequentially irradiated to obtain the first fluorescence information and the first excitation light.
  • the time for acquiring the first fluorescence information and the second fluorescence information can be shortened compared to the case of acquiring the second fluorescence information.
  • FIG. 21 is another example of the configuration of the measuring apparatus according to the second embodiment.
  • the measurement apparatus 10 may further include a separation unit 22 that separates the reflected light, the first fluorescence, and the second fluorescence between the irradiation target 1 and the second objective lens 21.
  • the separation unit may include a separation optical element (for example, a dichroic mirror) that separates reflected light and fluorescence.
  • the separation unit may include a bandpass filter (wavelength selection unit) that separates the first fluorescence and the second fluorescence.
  • the measuring apparatus 10 includes a third objective lens 23, a second microlens array 24, and a second sensor 25.
  • the second microlens array 24 and the second sensor 25 are arranged in this order in the vicinity of the focal plane of the third objective lens 23.
  • the configuration of the second microlens array 24 is the same as that of the microlens array 8.
  • the configuration of the second sensor 25 is the same as that of the sensor 5.
  • the separation unit 22 is configured to guide the reflected light and the first fluorescence to the sensor 5 side and guide the reflected light and the second fluorescence to the second sensor 25.
  • the signal string extraction unit 31 can calculate the reference focal position Z 0 from the set of pixel signals of the multifocal reflected light acquired by the sensors 5 and 25, respectively. For some reason, there may be a difference between the optical path length to the sensor 5 side and the optical path length to the second sensor 25 side. Even if the deviation in the optical path amounts occur, obtained by the use of the reference focus position Z 0 obtained from the information of the received reflected light in each sensor 5,25, without the influence of the deviation of the optical path length, the information of each fluorescent can do.
  • ⁇ Z 1 ⁇ Z 2 .
  • the measurement apparatus 10 may include a second separation unit (for example, a dichroic mirror) for separating the third fluorescence and the fourth fluorescence. Good.
  • a second separation unit for example, a dichroic mirror
  • the signal sequence at the focal point of the first fluorescence, and A signal string at the focal point of the second fluorescence can be acquired.
  • FIG. 22 is an example of a flowchart for measuring the irradiated object 1 by the measuring apparatus 10.
  • the irradiated object 1 (for example, one irradiated object among a plurality of irradiated objects 1 supported by the support member 60) is the first excitation light, the second excitation light, and the reference light. It is irradiated with three lights.
  • the light source device 2 emits the first excitation light, the reference light, and the second excitation light in the order of the first excitation light, the reference light, and the second excitation light.
  • FIG. 22 the irradiated object 1 (for example, one irradiated object among a plurality of irradiated objects 1 supported by the support member 60) is the first excitation light, the second excitation light, and the reference light. It is irradiated with three lights.
  • the light source device 2 emits the first excitation light, the reference light, and the second excitation light in the order of the first excitation light, the reference light
  • the period in which the light source device 2 emits the first excitation light, the period in which the light source device 2 emits the reference light, and the period in which the light source device 2 emits the second excitation light do not overlap. . Therefore, the irradiated object 1 is irradiated in the order of the first excitation light, the reference light, and the second excitation light.
  • the period in which the light source device 2 emits the reference light is between the period in which the light source device 2 emits the first excitation light and the period in which the light source device 2 emits the second excitation light. Note that the emission order of the first excitation light, the second excitation light, and the reference light is not limited to this, and may be any order.
  • Steps 2201 to 2209 in FIG. 22 include the same contents as 1001 and 1101 to 1108 in FIG.
  • the signal sequence extraction unit 31 determines a reference focal position from a set of pixel signals of reflected light at a plurality of different focal points (2210). As an example, the signal sequence extraction unit 31 generates an image from a signal sequence of pixel signals at a common focal position, and holds a plurality of images at a plurality of focal positions. The signal sequence extraction unit 31 performs the following processing using information of a plurality of images. Note that it is not always necessary to treat the image as an image, and the following processing may be performed with a set of pixel signals.
  • the signal sequence extraction unit 31 divides the image at each focal position into a plurality of regions.
  • FIG. 23 shows an example in which the images at the respective focal positions Z ⁇ 3 to Z 3 are divided into a plurality of regions.
  • the signal sequence extraction unit 31 may obtain the contrast at each focal position for all of the plurality of areas and determine the reference focal position for each area.
  • the signal string extraction unit 31 calculates the contrast of the region corresponding to the same position in the images at the respective focal positions Z ⁇ 3 to Z 3 .
  • the signal sequence extraction unit 31 determines a position where the contrast is maximum from among the focus positions Z ⁇ 3 to Z 3 as the reference focus position.
  • the signal sequence extraction unit 31 calculates the contrast for each region, and determines the reference focal position for each region.
  • FIG. 24 is an example of the inclination of the irradiated object 1.
  • the in-focus position is different between a region near the right end of the irradiated body 1 (a region including the first vertex 1a) and a region near the left end of the irradiated body 1 (a region including the third vertex 1c). . Therefore, the signal sequence extraction unit 31 determines the reference focal position for each region.
  • the signal sequence extraction unit 31 may obtain the contrast at each focal position with respect to at least three regions among a plurality of regions. This is because the inclination of the plane can be specified if at least three points are determined. As an example, the signal sequence extraction unit 31 may determine, for each region, the focal position where the contrast is maximum as the reference focal position.
  • the stage control unit 33 determines whether the signal values of the pixel signals of all the calculated reference focal positions (three reference focal positions) are larger than a predetermined threshold (2211).
  • the signal value is a luminance value.
  • the stage control unit 33 determines whether all the luminance values of the three reference focal positions exceed a predetermined threshold value.
  • the support member 60 may be regarded as being inclined at a certain level or more. If the inclination of the support member 60 is greater than or equal to a certain level, the reference focal position for each region differs greatly, and as a result, the signal sequence corresponding to the first fluorescent focal point and the second fluorescent focal point correspond to each other.
  • the process proceeds to step 2209.
  • the stage control unit 33 may drive the stage 4 and adjust the tilt of the stage 4 ( 2209). If the above condition is satisfied, the process proceeds to step 2212.
  • the stage control unit 33 may display an error on the display device 40 when the signal value of the pixel signal at the reference focal position is equal to or less than a predetermined threshold value.
  • the stage controller 33 may record the content of the error as data in the storage device when the signal value of the pixel signal at the reference focal position is equal to or less than a predetermined threshold value.
  • the stage control unit 33 calculates the contrast of the region corresponding to the calculated three reference focal positions, and determines whether all of the contrast values are larger than a predetermined threshold value. May be. If even one of the contrasts does not exceed the threshold value, the support member 60 may be regarded as being inclined more than a certain level.
  • the signal sequence extraction unit 31 determines the reference focal position with respect to three areas out of a plurality of areas, and determines the inclination of the irradiated object 1 from the reference focal position for each area. Good. Since various dimensions (vertical dimension, horizontal dimension, distance between spots, etc.) of the irradiation object 1 are known in advance, the signal string extraction unit 31 includes information on three reference focal positions and various dimensions of the irradiation object 1. May be used to calculate the inclination of the irradiated object 1. As an example, the stage control unit 33 may determine whether the calculated inclination is larger than a predetermined threshold value. As an example, the stage control unit 33 may change or adjust the height (Z direction) of the stage 4 when the calculated inclination is larger than a predetermined threshold value.
  • the signal sequence extraction unit 31 determines the reference focal position in all regions other than the three regions. As an example, if the three reference focal positions are the same, the signal sequence extraction unit 31 may determine all the areas other than the three areas as the same reference focal position. In this case, the subsequent steps 2112 to 2114 are the same as steps 1110 to 1112 in FIG. Therefore, the description is omitted.
  • the signal sequence extraction unit 31 may determine each peripheral region as the same reference focus position. For example, if the reference focal position in the area including the first vertex 1a of the irradiated object 1 is Z 0 , the signal sequence extraction unit 31 sets the reference focal positions of several areas around the area as Z 0 . You may judge. Referring focal position in the region including the second apex 1b of the irradiation object 1 is to Z 1, the signal sequence extraction unit 31 determines a reference focus position of some regions of the periphery of the area with Z 1 May be. Referring focal position in the region including the third apex 1c of the irradiation object 1 is to Z 2, the signal sequence extraction unit 31 determines a reference focus position of some regions of the periphery of the region and Z 2 May be.
  • the signal sequence extraction unit 31 determines reference focal positions in all regions other than the three regions according to the calculated inclination of the irradiated object 1.
  • the signal sequence extraction unit 31 obtains a change amount in the Z direction between adjacent regions according to the calculated inclination, and determines a reference focal position based on the change amount.
  • the value in the Z direction in each area calculated on the basis of the amount of change is less than or equal to the intermediate value of Z 0 and Z 1
  • a reference focus position determined to Z 0, the Z 0 and Z 1 larger than the intermediate value is a reference focus position may be determined to Z 1.
  • FIG. 25 is a diagram for explaining the processing in steps 2212 and 2213 when the reference focal positions are different in a plurality of regions of the irradiated object 1.
  • FIG. 25 shows a set of reflected light pixel signals, a set of first pixel signals, and a set of second pixel signals.
  • the signal string extraction unit 31 acquires information on the first fluorescence chromatic aberration amount ⁇ Z 1 from the chromatic aberration information storage unit 34.
  • the signal sequence extraction unit 31 uses the first fluorescence chromatic aberration amount ⁇ Z 1 from the reference focal position for each region of the irradiated object 1, and the signal sequence of the pixel signal corresponding to the focal point of the first fluorescence. Is extracted (2212).
  • a set of each pixel signal is acquired at a plurality of different focal points Z ⁇ 4 to Z 4 .
  • the reference focal position of a plurality of areas including the first vertex 1a of the irradiated object 1 is Z 0
  • the second vertex 1b of the irradiated object 1 is Referring focal position of a plurality of regions (central region of the irradiated object 1) containing it is determined that Z 1, a plurality of regions (left end in the region of the irradiation object 1 including a third apex 1c of the irradiation object 1 ) reference focal position in it is assumed to have been determined as Z 2.
  • the shaded portion 2501 in FIG. 25 indicates the reference focal position for each region.
  • the signal sequence extraction unit 31 For a plurality of areas including a first apex 1a of the irradiation object 1, the signal sequence extraction unit 31, the first fluorescence from the reference focus position Z 0 with chromatic aberration amount [Delta] Z 1, of the first pixel signal from the set, to acquire a signal sequence of the pixel signals of the focus position Z 2 (hatched portion of Z 2 of 2502) (arrow 2511).
  • the signal sequence extraction unit 31 For a plurality of regions including a second vertex 1b of the irradiation object 1, the signal sequence extraction unit 31, the first fluorescence from the reference focus position Z 1 using chromatic aberration amount [Delta] Z 1, of the first pixel signal from the set, to acquire a signal sequence of the pixel signals of the focus position Z 3 (shaded portion of Z 3 of 2502) (arrow 2512).
  • the signal sequence extraction unit 31 For a plurality of areas including a third apex 1c of the irradiation object 1, the signal sequence extraction unit 31, the first fluorescence from the reference focus position Z 2 using chromatic aberration amount [Delta] Z 1, of the first pixel signal from the set, to acquire a signal sequence of the pixel signals of the focus position Z 4 (shaded portion of Z 4 of 2502) (arrow 2513).
  • the combination of the signal sequences extracted as described above corresponds to a signal sequence representing a fluorescence image at the focal point of the first fluorescence.
  • the signal sequence extraction unit 31 acquires information on the second fluorescence chromatic aberration amount ⁇ Z 2 from the chromatic aberration information storage unit 34.
  • the signal string extraction unit 31 uses the second fluorescence chromatic aberration amount ⁇ Z 2 from the reference focal position for each region of the irradiated object 1, and the signal string of the pixel signal corresponding to the focal point of the second fluorescence. Is extracted (2213).
  • the signal sequence extraction unit 31 For a plurality of areas including a first apex 1a of the irradiation object 1, the signal sequence extraction unit 31, the second fluorescence from the reference focus position Z 0 with chromatic aberration amount [Delta] Z 2, of the second pixel signal From the set, the signal sequence of the pixel signal at the focal position Z- 2 (the shaded portion of Z- 2 in 2503) is acquired (arrow 2521).
  • the signal sequence extraction unit 31 For a plurality of regions including a second vertex 1b of the irradiation object 1, the signal sequence extraction unit 31, the first fluorescence from the reference focus position Z 1 using chromatic aberration amount [Delta] Z 2, of the second pixel signal From the set, a signal sequence of the pixel signal at the focal position Z ⁇ 1 (the shaded portion of Z ⁇ 1 in 2503) is acquired (arrow 2522).
  • the signal sequence extraction unit 31 For a plurality of areas including a third apex 1c of the irradiation object 1, the signal sequence extraction unit 31, the second fluorescence from the reference focus position Z 2 using chromatic aberration amount [Delta] Z 2, of the second pixel signal From the set, the signal sequence of the pixel signal at the focal position Z 0 (the shaded portion of Z 0 in 2503) is acquired (arrow 2523).
  • the combination of the signal sequences extracted as described above corresponds to a signal sequence representing a fluorescence image at the focal point of the second fluorescence.
  • the signal sequence processing unit 32 performs first signal processing on the signal sequence of the pixel signal corresponding to the extracted first fluorescence focal point, and extracts the extracted second fluorescence focal point. Second signal processing is executed for the signal sequence of pixel signals corresponding to (2214). Note that the content of step 2214 is the same as that of step 1112 in FIG.
  • FIG. 26 is another example of three points for calculating the reference focal position.
  • the three points for calculating the reference focal position may be defined by isosceles triangles connecting the two vertices of the irradiated object 1 and the midpoints of the sides of the irradiated object 1. If these three points are determined, the inclination of the plane can be specified.
  • the reference focus position of the two vertices of the irradiation object 1 is determined to Z 1
  • reference focus position at the midpoint of the irradiated body 1 side is determined to Z 0.
  • this embodiment may be executed in any of the flowcharts of FIG. 10, FIG. 16, and FIG.
  • the signal sequence at the focal point of the first fluorescence and the signal sequence at the focal point of the second fluorescence can be acquired while determining the inclination of the irradiated object 1. . It is also possible to automatically detect that the irradiated object 1 is inclined more than a certain level and adjust the inclination of the irradiated object 1. Conventionally, the inclination of the irradiated object 1 cannot be determined. Conventionally, since the fluorescence image is acquired in accordance with a specific focal position (that is, one focal position) using the AF function, when the irradiated body is inclined, all of the irradiated body is There was a possibility that information at the focal point could not be obtained in the area.
  • the tilt of the irradiated object 1 it is possible to determine the tilt of the irradiated object 1 and determine the focal point position for each region of the irradiated object 1. Even when the reference focal position is different in each region of the irradiated object 1, a signal sequence of pixel signals corresponding to the focal point can be acquired.
  • this embodiment is applicable also to the structure which radiates
  • the signal string extraction unit 31 obtains a change in contrast with respect to at least three regions of the irradiated object 1, What is necessary is just to determine a peak.
  • the signal sequence extraction unit 31 may determine one of the three peaks as the reference focal position using the chromatic aberration information.
  • the light source device 2 emits the first excitation light and the second excitation light without emitting the reference light.
  • the light source device 2 emits the first excitation light and the second excitation light in the order of the first excitation light and the second excitation light.
  • the emission order of the first excitation light and the second excitation light is not limited to this, and may be the order of the second excitation light and the first excitation light.
  • the signal string extraction unit 31 acquires from the sensor 5 a set of first pixel signals at a plurality of different focal points and a set of second pixel signals at a plurality of different focal points. Next, as an example, the signal string extraction unit 31 determines the focal point position of the first fluorescence from a set of first pixel signals at a plurality of different focal points. As an example, the signal sequence extraction unit 31 determines the focal position of the pixel signal having the highest luminance value as the focal point position of the first fluorescence from the set of first pixel signals at a plurality of different focal points. If a pixel signal having a sufficient luminance value is obtained, the focal position where the luminance value is maximum can be regarded as the focal point position of the first fluorescence. The signal sequence extraction unit 31 acquires a signal sequence of pixel signals corresponding to the in-focus position of the first fluorescence determined here.
  • the chromatic aberration information storage unit 34 includes information indicating the relationship of the chromatic aberration amount between the first fluorescence and the second fluorescence. Using this relationship, it is possible to obtain the in-focus position of the second fluorescence from the in-focus position of the first fluorescence.
  • the signal sequence extraction unit 31 uses the amount of chromatic aberration of the second fluorescence from the focus position of the first fluorescence to extract the signal sequence of the pixel signal corresponding to the focus position of the second fluorescence.
  • the signal sequence of the pixel signals extracted here substantially corresponds to a signal sequence that represents a fluorescence image at the focal point of the second fluorescence.
  • the signal string extraction unit 31 may determine that a pixel signal having a sufficient luminance value has not been obtained when the highest luminance value is smaller than a predetermined threshold value. Depending on the specimen, fluorescence may not be emitted from the irradiated object 1. Information that a pixel signal having a sufficient luminance value cannot be obtained can also be effective information for the user.
  • the signal sequence corresponding to the focal point of the first fluorescence and the focal point of the second fluorescence are not performed without performing focusing by the AF function.
  • a corresponding signal sequence can be acquired.
  • the signal sequence extraction unit 31 determines the focal point position of the second fluorescence from the set of pixel signals of the second fluorescence, and the first fluorescence from the focal point position of the second fluorescence.
  • the signal sequence of the first fluorescence pixel signal may be extracted using the amount of chromatic aberration.
  • the measurement apparatus 10 may provide a separation unit that separates the first fluorescence and the second fluorescence between the irradiated object 1 and the microlens array 8.
  • the measuring apparatus 10 may include a third objective lens, a second microlens array, and a second sensor.
  • the second microlens array and the second sensor are arranged in this order near the focal plane of the third objective lens.
  • the configuration of the second microlens array is the same as that of the microlens array 8.
  • the configuration of the second sensor is the same as that of the sensor 5.
  • the separation unit is configured to guide the first fluorescence to the sensor 5 side and guide the second fluorescence to the second sensor.
  • the signal sequence extraction unit 31 may obtain a contrast from a set of pixel signals at a plurality of different focal positions and determine two contrast peaks.
  • FIG. 27 is a diagram illustrating a measurement system (screening apparatus) including the measurement apparatus 10 according to the above-described embodiment.
  • the measurement system is a system that automatically performs the measurement method of the irradiated object 1 described above.
  • the measurement system 100 includes a pretreatment device (reaction device, bioassay device) 101, a transport device (plate loader) 102, and a measurement device 103.
  • the pretreatment apparatus 101 is a bioassay apparatus that prepares an object 1 to be measured.
  • the pretreatment apparatus 101 injects a specimen (target) containing a labeled target into the irradiated object 1 (biomolecule arranged in the spot), and is specific to the biomolecule and the target. It is an apparatus for carrying out the reaction.
  • the pretreatment device 101 includes a stage device that supports a support member 60 (for example, a plate), a dispensing device that includes a dispensing nozzle that injects a sample into each spot, and an irradiation target after the sample is injected.
  • a cleaning device for cleaning the body for cleaning the body.
  • the pretreatment device 101 may include a drying device that dries the irradiated object after cleaning.
  • the pre-processing apparatus 101 may be configured to process the support members 60 one by one or may be configured to process a plurality of sheets simultaneously.
  • the transport device 102 is a transport mechanism that transports the support member 60 from the pretreatment device 101 to the measurement device 103.
  • a known transfer robot device can be used as the transfer device 102.
  • the transport apparatus 102 unloads the support member 60 from the stage apparatus of the pretreatment apparatus 101 and loads it into the measurement apparatus 103.
  • the transport apparatus 102 may be provided with a mechanism for temporarily waiting for the support member 60 after cleaning.
  • the measuring apparatus 103 includes the measuring apparatus 10 according to the above-described embodiment.
  • the measuring device 103 measures the irradiated object 1 of the support member 60 disposed on the stage 4 by the transport device 102.
  • the measurement process by the measurement apparatus 103 is as described above.
  • the conveyance device 102 carries out the support member 60 whose measurement has been completed from the stage 4 and conveys it to a predetermined position.
  • the pretreatment (reaction process, bioassay) for the irradiated object 1 and the measurement process for the irradiated object 1 after the pretreatment (reaction process, bioassay) are continuously performed.
  • Molecular arrays can be screened.
  • the control device 30 may include a switching processing unit for switching the processes of the first to fourth embodiments described above.
  • the switching processing unit can select one of the processes in the first to fourth embodiments.
  • the processing of the control device 30 described above can also be realized by software program codes that realize these functions.
  • a storage medium in which the program code is recorded is provided to the system or apparatus, and the computer (or CPU or MPU) of the system or apparatus reads the program code stored in the storage medium.
  • the program code itself read from the storage medium realizes the functions of the above-described embodiments, and the program code itself and the storage medium storing the program code constitute the present invention.
  • a storage medium for supplying such program code for example, a flexible disk, CD-ROM, DVD-ROM, hard disk, optical disk, magneto-optical disk, CD-R, magnetic tape, nonvolatile memory card, ROM Etc. are used.
  • control lines and information lines are those that are considered necessary for the explanation, and not all control lines and information lines on the product are necessarily shown. All the components may be connected to each other.
  • stage control unit 34 ... chromatic aberration information storage unit, 40 DESCRIPTION OF SYMBOLS ... Display apparatus, 51 ... Control line, 60 ... Support member, 81 ... Micro lens, 100 ... Measurement system, 101 ... Pre-processing apparatus, 102 ... Conveyance apparatus, 103 ... Measurement apparatus

Abstract

L'objectif de la présente invention est de fournir un appareil de mesure, un système de mesure, un procédé de traitement de chaîne de signal, et un programme qui sont aptes à améliorer un débit. L'appareil de mesure comprend un dispositif source de lumière pour émettre une première lumière d'excitation et une seconde lumière d'excitation vers un objet à exposer, un réseau de microlentilles comprenant une pluralité de microlentilles agencées de manière bidimensionnelle, un capteur qui a des éléments de réception de lumière agencés de façon bidimensionnelle, et qui reçoit, par l'intermédiaire du réseau de microlentilles, une première lumière fluorescente générée lorsque la première lumière d'excitation est émise sur l'objet à exposer, et une seconde lumière fluorescente générée lorsque la seconde lumière d'excitation est émise sur l'objet à exposer, et une unité de commande qui génère un premier résultat à partir de la chaîne de signal d'un premier signal de pixel correspondant au point focal de la première lumière fluorescente, et qui génère un second résultat à partir de la chaîne de signal d'un second signal de pixel correspondant au point focal de la seconde lumière fluorescente.
PCT/JP2015/060280 2015-03-31 2015-03-31 Appareil de mesure, système de mesure, procédé de traitement de chaîne de signal, et programme WO2016157458A1 (fr)

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