WO2014046138A1 - Sample analysis device, sample analysis method, sample analysis program, and particle track analysis device - Google Patents

Abstract

A concavity detection unit (514b) detects, as a recess recessed from an imaging surface, a portion (203d, 201d) at which the luminance value of an approaching image (203c) taken in a state in which the focal position of an objective lens (31) has moved to an approaching position is large and the luminance value of a separation image (201c) taken in a state in which the focal position has moved to a separation position is small. A convexity detection unit (513a) detects, as a protrusion projecting from the imaging surface, a portion (201b, 203b) at which the luminance value of the separation image (201a) is large and the luminance value of the approach image (203a) is small.

Description

Sample analyzer, sample analysis method, sample analysis program, and particle track analyzer

The present invention relates to a sample analysis device, a sample analysis method, a sample analysis program, and a particle track analysis device.

For example, Patent Documents 1 and 2 disclose techniques for detecting scratches and attached dust on a sample such as a film with a microscope, a scanner, a sensor, or the like.

JP 2003-232746 A Japanese Patent Laid-Open No. 11-215319

Although the techniques described in Patent Documents 1 and 2 can detect whether a sample has a concave portion (a scratch on the sample) or a convex portion (dust attached to the sample), the concave and convex portions of the sample are detected. Cannot be distinguished.

The present invention has been made in view of the above-described circumstances, and an object thereof is to detect a concave portion and a convex portion of a sample and discriminate between the concave portion and the convex portion.

In order to achieve the above object, a sample analyzer according to the first aspect of the present invention provides:
An emission part for emitting visible light;
A support part that is arranged on the optical axis of visible light emitted by the emission part and supports a sample through which visible light is transmitted;
An imaging unit that has an optical system and captures an image of the sample supported by the support unit;
The focal position of the optical system is set along the optical axis of visible light, the approach position closer to the emitting portion than the reference position of the imaging surface, which is the surface of the sample on the imaging portion side, and the emitting position than the reference position. A focal point moving part that is moved between a remote position away from the part,
The focus moving unit moves the focus position to the approach position, and the brightness value of the approach image that is the image of the sample captured by the imaging unit at the approach position is larger than the brightness value of the peripheral portion that is set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position by the focus moving unit moving the focal position to the remote position, is smaller than the luminance value of the peripheral portion. Concave detecting means for detecting a portion as a concave recessed from the imaging surface;
A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. Detection means;
It is characterized by providing.

In the above sample analyzer,
The approach / separation difference image obtained by subtracting the brightness value of each pixel corresponding to the separation image from the brightness value of each pixel of the approach image, and the brightness of each pixel corresponding to the approach image from the brightness value of each pixel of the separation image A difference acquisition means for acquiring a distance approaching difference image obtained by subtracting a value;
A closed region extracting means for extracting a closed region whose luminance value is different from the peripheral portion from the image;
Further comprising
The closed region extracting means moves the focus position to a measurement position moved by a preset measurement distance from the reference position along the optical axis, and the imaging unit images at the measurement position. A closed region in the measurement image as the image of the sample, a closed region in which the luminance value in the approaching / separating difference image is a positive value, and a closed region in which the luminance value in the separating / approaching difference image is a positive value. Extract the region and
The concave detecting means detects, as the concave portion, a closed region in the measurement image that is arranged at a position of the closed region where the brightness value in the approaching / separating difference image is a positive value,
The convex detecting means detects, as the convex portion, a closed region in the measurement image that is arranged at a position of the closed region where the luminance value in the separation approach difference image is a positive value. May be.

In the above sample analyzer,
When the closed region extraction unit extracts an inner boundary and an outer boundary surrounding the inner boundary as a boundary surrounding a portion having a luminance value different from other portions from the measurement image, The boundary of the region having the lowest average luminance value may be the boundary of the closed region in the measurement image.

In order to achieve the above object, a sample analysis method according to the second aspect of the present invention includes:
An imaging step of capturing an image of the sample by an imaging unit having an optical system when visible light is transmitted through the sample disposed on the optical axis;
The focal position of the optical system is closer to the light source than the reference position of the imaging surface, which is the surface on the imaging unit side of the sample, along the optical axis of visible light, and away from the light source than the reference position A focal point moving step for moving between the separated position and the focal point moving unit;
The focus moving unit moves the focus position to the approach position, and the brightness value of the approach image that is the image of the sample captured by the imaging unit at the approach position is larger than the brightness value of the peripheral portion that is set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position by the focus moving unit moving the focal position to the remote position, is smaller than the luminance value of the peripheral portion. A concave detecting step of detecting a portion as a concave recessed from the imaging surface;
A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. A detection process;
It is characterized by providing.

In order to achieve the above object, a sample analysis program according to the third aspect of the present invention provides:
Computer
An imaging unit that captures an image of the sample by an imaging unit having an optical system when visible light passes through the sample disposed on the optical axis;
The focal position of the optical system is closer to the light source than the reference position of the imaging surface, which is the surface on the imaging unit side of the sample, along the optical axis of visible light, and away from the light source than the reference position A focal point moving means for moving between the separated position and
The focus moving means moves the focus position to the approach position, and the brightness value of the approach image, which is the image of the sample imaged by the imaging unit at the approach position, is larger than the brightness value of the peripheral portion set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position with the focus moving means moving the focus position to the remote position, is smaller than the luminance value of the peripheral portion. Concave detecting means for detecting a portion as a concave recessed from the imaging surface;
A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. Detection means,
It is made to function as.

In order to achieve the above object, a particle track analysis apparatus according to the fourth aspect of the present invention provides:
An emission part for emitting visible light;
A support part that is disposed on the optical axis of visible light emitted by the emission part and supports a track detection solid that transmits visible light; and
An imaging unit that has an optical system and captures an image of a track detection solid as a sample supported by the support;
The focal position of the optical system is closer to the emitting part than the reference position of the imaging surface, which is the surface on the imaging part side of the solid for track detection, along the optical axis of visible light, and from the reference position And a focal point moving unit that moves between a remote position away from the emitting unit,
The brightness value of the peripheral portion in which the brightness value of the separated image, which is an image of the track detection solid image captured by the imaging unit at the separated position by the focus moving unit moving the focal position to the separated position, is set in advance. And the brightness value of the approach image, which is an image of the track detection solid image captured by the imaging unit at the approach position when the focus moving unit moves the focus position to the approach position, A foreign matter removing means for removing a portion smaller than the brightness value from the image of the track detection solid as a foreign matter attached to the imaging surface and protruding from the imaging surface;
A track of particles formed by recessing a portion where the luminance value of the approaching image is larger than the luminance value of the peripheral portion and the luminance value of the separated image is smaller than the luminance value of the peripheral portion from the imaging surface Track extraction means for extracting from the track detection solid image as,
It is characterized by providing.

According to the present invention, the concave and convex portions of the sample can be detected and the concave and convex portions can be distinguished.

It is a front view of the sample analyzer which concerns on this embodiment. It is a side view of a microscope part. It is a figure which shows the relationship between the visual field of a microscope, and the imaging range of an image sensor. It is a figure which shows the structural example of a Z-axis control part. It is a functional block diagram of a sample analyzer. (A) is a diagram showing a state in which the focal position of the objective lens is moved to the separation position, (B) is a diagram showing a state in which the focal position of the objective lens is moved to the measurement position, and (C) is a diagram showing the focal position of the objective lens. The figure which shows the state which moved to the approach position, (D) is a figure which shows a separated image, (E) is a figure which shows a measured image, (F) is a figure which shows an approach image, (G) shows an approached and separated difference image. FIG. 5H is a diagram showing a separation approach difference image. It is explanatory drawing of the target which has a multiple boundary. (A) is a figure which shows the state which extracted the target with high coincidence with the track | truck in the approaching / separating difference image, (B) is the state which extracted the target with high matching with the foreign material in the separating / separating difference image FIG. 8C is a diagram taken in a state where the focal position is moved to the measurement position. 6A is a diagram illustrating a state in which foreign matter is attached to the imaging surface in the state of FIG. 6A, FIG. 6B is a diagram illustrating a separated image captured in the state of FIG. 6A, and FIG. 6A is a diagram illustrating a state in which a track of particles is formed on the imaging surface in the state of FIG. 6A, FIG. 6D is a diagram illustrating a remote image captured in the state of FIG. 6C, and FIG. ) Is a diagram illustrating a state in which foreign matter is attached to the imaging surface, (F) is a diagram illustrating an approach image captured in the state of (E), and (G) is captured in the state of FIG. 6 (C). The figure which shows the state in which the track of a particle is formed on the surface, (H) is a figure which shows the approach image imaged in the state of (G). It is a flowchart of a sample analysis process. It is a flowchart of the target object extraction process of a measurement image. It is a flowchart of the target object extraction process of each difference image. It is a flowchart of a track extraction process. It is a figure which shows the measurement image by which the binarization process was carried out. It is a block diagram which shows an example of the hardware constitutions of a control computer.

Hereinafter, a sample analysis apparatus (particle track analysis apparatus) 100 according to an embodiment of the present invention will be described with reference to FIGS.

The sample analyzer 100 includes a microscope unit 110 and a control computer 150 as shown in FIG.

As shown in FIG. 2, the microscope unit 110 includes a moving unit 2 that movably supports the sample 1, a microscope 3 that magnifies the image of the sample 1, and an image sensor that captures an image of the sample magnified by the microscope 3. 4. The moving unit 2 and the microscope 3 are supported by an L-shaped gantry 7.

Sample 1 is a track detection solid. The track detection solid is an organic plastic plate. When the radiation passes through the track detection solid, the polymer bond is damaged in the portion through which the radiation has passed. When this damaged portion is etched with a predetermined solution, a minute recess called an etch pit is generated. This etch pit shows a track of radiation. The etch pit has a different shape (for example, a perfect circle or an ellipse), a diameter, and a depth depending on an incident amount and an incident direction of radiation. For this reason, by observing the number and shape of the etch pits generated in the sample 1 with a microscope, it is possible to acquire information such as the amount of incident radiation and the incident direction.

Dust may adhere to the surface of the sample 1. When an image of the sample 1 to which dust is attached is captured, it is necessary to discriminate between etch pits and dust in the captured image.

The moving part 2 is installed in the horizontal part of the L-shaped gantry 7. The moving unit 2 includes an XY stage 21, a sample stage (supporting unit) 22, a condenser lens 23, and an encoder 24.

The sample stage 22 supports the sample 1. The sample stage 22 is installed on the XY stage 21.
The XY stage 21 is driven by an X-axis motor 131 and a Y-axis motor 132 shown in FIG. 5, which will be described later, and moves horizontally in the left-right and front-back directions (X-axis direction and Y-axis direction). As the XY stage 21 moves, the sample stage 22 and the sample 1 supported on the sample stage 22 also move. The X-axis motor 131 and the Y-axis motor 132 are controlled by the control computer 150.
The condenser lens 23 is installed under the sample table 22.
The encoder 24 supplies the control computer 150 with pulses corresponding to the amounts of movement of the XY stage 21 in the X-axis direction and the Y-axis direction. The encoder 24 includes an X-axis encoder 24X and a Y-axis encoder 24Y. Position correction (shift correction) in the X-axis direction and the Y-axis direction with respect to an area image acquired by the image sensor 4 described later is performed based on outputs from the X-axis encoder 24X and the Y-axis encoder 24Y, respectively.

The microscope 3 includes an objective lens (optical system) 31, an illumination unit 32 that irradiates the sample 1, a control imaging unit 33, a lens barrel 34, an eyepiece 35 for visual observation, and a Z-axis control unit (focus). Moving part) 36.

The objective lens 31 includes a plurality of lenses having different magnifications (for example, a lens having a magnification of 10 times and a lens having a magnification of 20 times). The plurality of lenses can be switched by rotating the revolver 37.

The illumination unit 32 includes an epi-illumination unit 32a and a transmission illumination unit 32b.
The epi-illumination unit 32 a irradiates the sample 1 with the light emitted from the built-in light source being bent along the optical axis of the microscope 3. At this time, the reflected light from the sample 1 enters the objective lens 31.
The transmitted illumination unit 32 b is provided in the horizontal portion of the L-shaped mount 7 under the sample table 22. The transmitted illumination unit 32b irradiates the sample 1 with light emitted from a light source built in the transmitted illumination unit 32b or light emitted from an external light source introduced from the optical fiber 8 from below. At this time, the transmitted light from the sample 1 enters the objective lens 31. In the present embodiment, the light irradiated to the sample 1 from the transmission illumination unit 32b is collected by the condenser lens 23 and irradiated (emitted) to the sample 1 as parallel light (collected parallel light). For this reason, the transmitted light from the sample 1 is parallel light.
The transmitted illumination unit 32b and the condenser lens 23 constitute an output unit 41 as shown in FIG. 2 and FIG. 9 described later.

The lens barrel 34 supports an eyepiece 35 for visual observation and the image sensor 4. A side portion of the lens barrel 34 is attached to an upright portion of the L-shaped gantry 7 via a Z-axis control unit 36.

The Z-axis control unit 36 moves the position of the microscope 3 in the vertical direction (Z-axis direction, optical axis direction). The Z-axis control unit 36 includes a slide plate 36 a fixed to the gantry 7, a slide plate 36 b fixed to the lens barrel 34, and a Z-axis motor 361.

As shown in FIG. 4, one of the slide plates 36 a and 36 b includes a Z-axis motor 361, a pinion gear 363 fixed to the rotation shaft 362 of the Z-axis motor 361, and an operation configured to be coaxial with the rotation shaft 362. And a knob 364.
The other of the slide plates 36 a and 36 b includes a rack 365. A rack and pinion mechanism is configured by engaging a pinion gear 363 with the rack 365. The pinion gear 363 rotates by driving the Z-axis motor 361 and the operation knob 364 to drive the rack 365. When the rack 365 is driven, the slide plate 36b slides in the vertical direction with respect to the slide plate 36a fixed to the gantry 7, and the lens barrel 34 moves in the vertical direction (Z-axis direction).

The eyepiece 35 for visual observation makes it possible to visually observe the sample 1 by tilting the optical axis of the light incident from the objective lens 31 with a prism.

The image sensor 4 is housed in a case and is detachably attached to the tip of the lens barrel 34. For example, a C mount or an F mount is used as the mounting portion.
The image sensor 4 includes CCD (Charge Coupled Device) elements arranged on the XY plane.

As shown in FIG. 3, the image sensor 4 captures an area included in the area AR that is the imaging range of the image sensor 4 in the image of the sample 1 captured in the visual field VF of the microscope 3. The image sensor 4 sequentially images the sample 1 that is moved in the X-axis direction or the Y-axis direction by the moving unit 2 for each area AR that is an imaging range, and connects the image data (area image data) of each area AR. This is transmitted to the control computer 150 via the code. That is, the image sensor 4 scans the image of the sample 1 magnified by the microscope 3 by capturing the image while performing relative movement with respect to the sample 1, and supplies the captured area image data to the control computer 150.

The control computer 150 is, for example, a computer.
As illustrated in FIG. 1, the control computer 150 includes an arithmetic processing unit 51, a display unit 52, and an image recording device 53.
The arithmetic processing unit 51 sets an imaging region, movement control and position control of the moving unit 2 in the X-axis direction and the Y-axis direction, movement control of the microscope 3 in the Z-axis direction, imaging control of the image sensor 4, and image sensor 4 The area image data supplied from the image data is fetched, and the entire image of the imaging area is created based on the area image data.

FIG. 5 shows a functional block diagram centering on the arithmetic processing unit 51.
As shown in FIG. 5, the arithmetic processing unit 51 is connected to the moving unit 2, the Z-axis control unit 36, the imaging unit 123, the image RAM 124, the image recording device 53, the data communication unit 125, and the like.

The moving unit 2 includes an X-axis drive unit 121, a Y-axis drive unit 122, an X-axis encoder 24X, and a Y-axis encoder 24Y.
The X-axis drive unit 121 drives the X-axis motor 131 according to the control of the arithmetic processing unit 51 to move the XY stage 21 in the X-axis direction.
The Y-axis drive unit 122 drives the Y-axis motor 132 according to the control of the arithmetic processing unit 51, and moves the XY stage 21 in the Y-axis direction orthogonal to the X-axis direction.

The X-axis encoder 24X is composed of a linear encoder or the like, and outputs a pulse train corresponding to the moving direction and distance when the XY stage 21 moves in the X-axis direction. The arithmetic processing unit 51 determines the moving direction and moving distance of the XY stage 21 in the X-axis direction by determining the number of pulses, the phase, and the like.

The Y-axis encoder 24Y is composed of a linear encoder or the like, and outputs a pulse train corresponding to the moving direction and distance when the XY stage 21 moves in the Y-axis direction. The arithmetic processing unit 51 determines the moving direction and moving distance of the XY stage 21 in the Y-axis direction by determining the number of pulses, the phase, and the like.

The Z-axis control unit 36 includes the aforementioned Z-axis motor 361, a Z-axis drive unit 366, and a Z-axis encoder 367. The Z-axis drive unit 366 drives the Z-axis motor 361 according to the control of the arithmetic processing unit 51 to move the microscope 3 in the Z-axis direction (vertical direction). The Z-axis encoder 367 is composed of a linear encoder or the like, and outputs a pulse train corresponding to the moving direction and distance when the microscope 3 moves in the Z-axis direction. The arithmetic processing unit 51 determines the moving direction and moving distance of the microscope 3 by determining the number of pulses, the phase, and the like.

The Z-axis control unit 36 detects the contrast of the image acquired by the image sensor 4 and performs autofocus processing, whereby the imaging surface (sample surface, imaging target surface) that is the surface of the sample 1 on the side close to the objective lens 31. And the focal position of the objective lens 31 included in the imaging unit 123 can be moved to the imaging surface. In the present specification, the focal position of the objective lens 31 in a state where the focal position of the objective lens 31 coincides with the imaging surface is referred to as a reference position.
Further, the Z-axis control unit 36 sets the focal position of the objective lens 31 to a separation position that is separated from the reference position by a preset separation distance (on the side closer to the objective lens) as shown in FIG. FIG. 6B shows a measurement position that is separated from the reference position by a preset measurement distance, and FIG. 6C shows a measurement position that is below the reference position by a preset approach distance (far from the objective lens). Between the approach positions separated from each other on the basis of the output signal of the arithmetic processing unit 51. In the present embodiment, the focal position of the objective lens 31 in a state where the focal position of the objective lens 31 is in a portion recessed from the imaging surface of the sample 1 rather than the approach position (a focal position with a high sharpness, a just-focused position). Position) is set as the measurement position.
In the present embodiment, the separation position and the approach position are set based on the thickness of the sample 1, the assumed particle track, the size of dust attached to the sample 1, and the like.

The imaging unit 123 includes the objective lens 31, the image sensor 4, and the like. Based on the output signal of the arithmetic processing unit 51, the imaging unit 123 captures a plurality of images of the sample 1 whose focal positions of the objective lens 31 are different from each other when imaging each image, and supplies the images to the arithmetic processing unit 51. Specifically, the imaging unit 123 captures the separated image (Far image) 201 shown in FIG. 6D captured in a state where the focal position is aligned with the separated position, and the focused position is aligned with the measured position. The measured image (Msr: Measure image) 202 shown in FIG. 6 (E) and the approach image (Near image) 203 shown in FIG. 6 (F) captured in a state where the focus position is adjusted to the approach position. An image is taken and supplied to the arithmetic processing unit 51.

The image RAM 124 functions as a work area for the arithmetic processing unit 51.

The image recording device 53 is composed of a mass storage device or the like, and records captured images.
The data communication unit 125 transmits / receives various data to / from an external device.

The arithmetic processing unit 51 includes a difference acquisition unit 511, a target object extraction unit 512, a foreign matter removal unit 513, and a track extraction unit 514.

The difference acquisition unit 511 obtains a close-up difference image (Near-Far image, a concave image) using a luminance value of each pixel as a value obtained by subtracting the luminance value of each corresponding pixel of the separate image 201 from the luminance value of each pixel of the close-up image 203. A difference image for detection (see FIG. 6G) 204 is acquired. In addition, the difference acquisition unit 511 uses a value obtained by subtracting the luminance value of each corresponding pixel of the approach image 203 from the luminance value of each pixel of the remote image 201 as a luminance value of each pixel (Far-Near image). , A difference image for convex detection (see FIG. 6H) 205 is acquired. The difference acquisition unit 511 of the present embodiment performs smoothing processing on the approach image 203 and the separation image 201 to smooth the contour lines in the image before acquiring the approach / separation difference image 204 and the separation approach difference image 205. Yes.
It should be noted that the difference images 204 and 205 shown in FIGS. 6G and 6H are different from black only in a portion where the luminance value has a positive value (positive information) for easy understanding. Is displayed.

The target object extraction unit (closed region extraction unit) 512 obtains, from each image, a portion (Obj: Object, closed region) whose luminance value is different from other portions (a white portion or a black portion than other portions). Extract as The target object extraction unit 512 includes a target object in the measurement image 202 shown in FIG. 6E, a target object whose luminance value in the approaching / separating difference image 204 shown in FIG. A target whose luminance value in the separated approach difference image 205 shown in (H) has positive information is extracted.

When extracting the target in the measurement image 202, the target extraction unit 512 first performs a smoothing process on the measurement image 202, and in the measurement image 202, a boundary that surrounds a portion whose luminance value is different from other portions ( Clarify the edges. Next, the target object extraction unit 512 executes a boundary extraction process (edge extraction process) for extracting boundaries in the measurement image 202, and labels the connected boundaries one by one.

By the way, in an image in which the same target is photographed at a plurality of different focal positions, due to light refraction, there is only one boundary in imaging at one focal position, but multiple boundaries ( (Multiple circles) may occur. Alternatively, multiple boundaries may occur for the same target in imaging at all focal positions. The target object extraction unit 512 is unnecessary when an outer boundary (outer boundary) surrounding the inner boundary (inner boundary) exists, that is, when a target having multiple boundaries (multiple circles) is extracted. Execute boundary removal processing to remove the boundary. Specifically, among the boundaries, an average luminance value (average value of luminance values) inside the boundary is measured, and a boundary having the lowest average luminance value in the boundary is extracted as a target boundary in the measurement image 202. And remove the remaining boundaries.
For example, the target 202a in the measurement image 202 shown in FIG. 7 has an outer boundary 202b, an intermediate boundary 202c, and an inner boundary 202d, as shown by the broken line in FIG. Therefore, the target object extraction unit 512 first calculates the average luminance value of the region in the boundary 202b, the region in the boundary 202c, and the region in the boundary 202d. At this time, the average luminance value is the entire region inside the boundary. Specifically, when the average luminance value of the region within the boundary 202c is obtained, the average luminance value of the region within the region 202c is calculated together with the luminance value of the region within the boundary 202d. In the case of the target 202a, the average luminance value of the region in the intermediate boundary 202c is the lowest, so the boundary of the target 202a becomes the intermediate boundary 202c, and the remaining boundaries 202b and 202d are removed.
If the average luminance value of the area in the intermediate boundary 202c and the average luminance value of the area in the inner boundary 202d are the same value, the intermediate boundary outside the inner boundary 202d. 202c becomes the boundary of the target 202a, and the inner boundary 202d is removed.

In addition, when the target object extraction unit 512 extracts a target object whose luminance value in each of the difference images 204 and 205 has positive information, first, the light amount of each pixel in the measurement image 202 has a preset threshold value. Whether or not to exceed is determined, and based on the determination result, a binarization extraction process for binarizing the luminance value of each pixel into a white value and a black value is executed. Next, the target object extraction unit 512 labels a set of pixels each having a luminance value of a white value connected in units of pixels one by one. Note that the purpose of this embodiment is to remove foreign matters and extract the tracks of particles, so the target discharge unit 512 is a technique called elliptic fitting described in JP-A-2005-293299. Is used to exclude from the target a set of labeled pixels that is not considered a track or a foreign object.

Specifically, the target extraction unit 512 creates an approximate ellipse that approximates the outline of the pixel set based on the coordinates of each pixel that forms the boundary of the labeled pixel set. Next, the target extraction unit 512 measures the shortest distance to the outer periphery (boundary) of the approximate ellipse existing in the vicinity of the pixel coordinates indicating the boundary. For example, when the shortest distance is within a preset threshold value It is determined as a valid pixel. Next, when the number of pixels determined to be valid reaches a predetermined threshold or more, the target extraction unit 512 determines that the degree of coincidence with a track or a foreign object is high, and extracts the pixel set as a target. To do. In addition, when the target extraction unit 512 determines that the number of pixels determined to be valid does not reach a predetermined threshold and the degree of matching is low, the target extraction unit 512 excludes the pixel set from the target.

For example, when binarization processing is performed on each of the difference images 204a and 205a shown in FIGS. 8A and 8B, the part surrounded by the solid line in FIGS. 8A and 8B is , The degree of match exceeds the threshold value and is extracted as a target. On the other hand, the portion surrounded by the dotted lines in FIGS. 8A and 8B has a matching degree equal to or less than the threshold value and is excluded from the target.
Further, in the present embodiment, after acquiring the barycentric coordinates of the target whose luminance value has positive information in each extracted difference image 204, 205, the luminance value in each difference image 204, 205 is positive information. The extraction process of the target having

The foreign matter removing unit 513 removes foreign matter attached to the imaging surface from the measurement image 202. The foreign substance removal unit 513 includes a convex detection unit 513a.

Here, when the foreign matter 1a shown in FIGS. 9A and 9E is attached to the imaging surface, the foreign matter 1a is irradiated as shown by the solid lines in FIGS. 9A and 9E. The focused parallel light is refracted and the focal point is formed above the imaging surface. That is, the foreign material 1a functions like a convex lens.
For this reason, when the focal position of the objective lens 31 is moved to the separation position, as shown by the broken line in FIG. 9A, the focal position of the objective lens 31 and the focal position of the transmitted light in the portion corresponding to the foreign matter 1a are obtained. Nearly, the amount of light increases. For this reason, in the separated image 201a shown in FIG. 9B, the luminance value of the portion 201b corresponding to the foreign object 1a is larger than the luminance value of the peripheral portion (close to white).
When the focal position of the objective lens 31 is moved to the approach position, as shown by the broken line in FIG. 9E, the focal position of the objective lens 31 and the focal position of the transmitted light corresponding to the foreign matter 1a are far away. , Less light. Therefore, in the captured approach image 203a shown in FIG. 9F, the luminance value of the portion 203b corresponding to the foreign object 1a is smaller than the luminance value of the peripheral portion (close to black).

As a result, the foreign matter 1a in which the portions 201b and 203b in which the luminance value in the separated image 201a is larger than the luminance value in the peripheral portion and the luminance value in the approach image 203a is smaller than the luminance value in the peripheral portion are attached to the imaging surface. Can be considered and removed. Therefore, the foreign matter removing unit 513 detects the convex portion protruding from the imaging surface by the convex detecting portion 513a described later, and removes the detected convex portion as the foreign matter 1a.

The convex detection unit 513a projects from the imaging surface portions 201b and 203b in which the luminance value in the separated image 201 is larger than the luminance value in the peripheral portion and the luminance value in the approaching image 203 is smaller than the luminance value in the peripheral portion. It detects as a convex part.
For example, when the convex detection unit 513a detects a convex part from the separation approach difference images 205 and 205a shown in FIGS. 6H and 8B, the brightness value of the positive information is detected in the separation approach difference images 205 and 205a. All the obtained targets (removal Obj) are detected as convex portions.

The track extraction unit 514 extracts a track of particles formed to be recessed from the imaging surface. The track extraction unit 514 includes a concave detection unit 514a and a non-extraction target removal unit 514b.

Here, when the particle tracks 1b shown in FIGS. 9C and 9G are formed on the imaging surface, as shown by the solid lines in FIGS. 9C and 9G, The parallel light applied to the track 1b is refracted and scattered, and a focal point is formed below the imaging surface. That is, the particle track 1b functions like a concave lens.
For this reason, when the focal position of the objective lens 31 is moved to the separation position, as shown by the broken line in FIG. 9C, the focal position of the objective lens 31 and the focal position of the transmitted light in the part corresponding to the track 1b of the particles. Is far away and the amount of light is small. For this reason, in the separated image 201c shown in FIG. 9D, the luminance value of the portion 201d corresponding to the particle track 1b is smaller than the luminance value of the peripheral portion. When the focal position of the objective lens 31 is moved to the approach position, as shown by a broken line in FIG. 9G, the focal position of the objective lens 31 and the focal position of the transmitted light in the portion corresponding to the particle track 1b Is close and there is a lot of light. For this reason, in the approach image 203c shown in FIG. 9 (H), the luminance value of the portion 203d corresponding to the particle track 1b is larger than the luminance value of the peripheral portion. Therefore, when the particle track 1b is formed on the imaging surface, an image in which the brightness and darkness of the corresponding portion is reversed is obtained compared to the case where the foreign matter 1a is attached to the imaging surface (an image with black and white reversed is obtained). ).

As a result, the portions 203d and 201d in which the luminance value in the approach image 203c is larger than the luminance value in the peripheral portion and the luminance value in the separated image 201c is smaller than the luminance value in the peripheral portion It can be extracted as a track 1b. Therefore, the track extraction unit 514 detects a recess recessed from the imaging surface by a recess detection unit 514a described later, and extracts the detected recess as a track 1b of particles.
Further, in this embodiment, since the purpose is to extract the particle track 1b, the track extraction unit 514 is different from the particle track 1b in the measurement image 202 by a non-extraction target removal unit 514b described later. The target is discriminated based on the boundary length (edge length), area, etc., and removed from the extraction target.

The concave detection unit 514a dents portions 203d and 201d from the imaging surface whose luminance value in the approach image 203c is larger than the luminance value in the peripheral portion and whose luminance value in the separated image 201c is smaller than the luminance value in the peripheral portion. Detect as a concave.
For example, when the concave detection unit 514a detects a concave portion from the approach / separation difference images 204, 204a shown in FIGS. 6 (G) and 8 (A), the luminance value of the positive information is obtained in the approach / separation difference images 204, 204a. All the target objects (extracted Obj) are detected as concave portions.

The non-extraction target removing unit 514b removes a target in the measurement image 202 that is different from the particle track 1b as a non-extraction target.
The non-extraction target removal unit 514b removes the target in the measurement image 202 whose boundary length is shorter than the preset threshold value for boundary LL.
Further, the non-extraction target removal unit 514b has a target area in the measurement image 202 whose area is smaller than the preset area minimum value SL and greater than the preset area maximum value SH. Remove things and things.
In the present embodiment, the area of the target in the measurement image 202 is obtained by obtaining the area of the ellipse obtained by the ellipse fitting process for obtaining an ellipse that approximates the outline of the target using a predetermined number of pixels extracted from the boundary. Ask for.
Further, the non-extraction target removal unit 514b excludes the target in the measurement image 202 that has a value obtained by dividing the obtained minor axis of the ellipse by the major axis is smaller than the preset ellipse threshold OD.

Note that the track extraction unit 514 of the present embodiment prevents the extraction of the tracks of the particles to be extracted (prevents counting leakage), so that a track larger than a predetermined size included in the measurement image 202 is the difference image. 204 and 205 are unconditionally extracted without collation. Specifically, the track extraction unit 514 is larger than a preset area threshold value SE among targets in the measurement image 202 whose area is not less than the area minimum value SL and not more than the area maximum value SH. Is extracted as a track of particles without matching with a target having a luminance value of positive information in the approaching / separating difference image 204 (unconditionally extracted).

Further, the track extraction unit 514 of the present embodiment obtains the center-of-gravity coordinates of the target in the measurement image 202, and the center-of-gravity coordinates of the target having the luminance value of the positive information in the approaching / separating difference image 204 Is obtained, and this value is stored as the distance for the track.
Next, the track extraction unit 514 obtains the distance between the center of gravity coordinates of the target in the obtained measurement image 202 and the center of gravity coordinates of the target having the positive brightness value in the separation approach difference image 205. The value is stored as the distance for the foreign object.
The track extraction unit 514 has a shorter track distance than the preset distance threshold GD, and all the determined foreign object distances are greater than or equal to the distance threshold GD. Then, the target in the measurement image 202 is extracted as a track of particles. The distance threshold GD is set to a value that is larger than any of all the obtained distances for tracks and smaller than any of the obtained distances for all foreign objects.

For example, a case where a measurement image 1202a in FIG. 8C is a target will be described.
First, the track extraction unit 514 obtains the center-of-gravity coordinates COD 1202a of the target 1202a. Next, the track extraction unit 514 obtains, for example, the center-of-gravity coordinates COD1202b of 1202b as the center-of-gravity coordinates of the target having the luminance value of the positive information in FIG. The distance between them is obtained, and the obtained distance is set as a track distance GD1.
Next, the track extraction unit 514 obtains, for example, the centroid coordinates COD1202c of 1202c as the centroid coordinates of the target having the luminance value of the positive information in FIG. 8B, which is the separation approach difference image, and between the COD1202a and the COD1202c. And the obtained distance is defined as a foreign object distance GD2.

In this case, the center-of-gravity coordinates COD 1202a of the target 1202a in the measurement image and the center-of-gravity coordinates COD 1202b of the target 1202b in the approach / separation difference image match if there is no measurement error. For this reason, the track distance GD1 is very small and is not more than the distance threshold GD. Therefore, it is determined that there is an object closer than the distance threshold GD in the approach / separation difference image.

On the other hand, in FIG. 8B, which is an isolated approach difference image, the target corresponding to the target 1202a is excluded from the object, and the closest target is 1202c. In this case, the foreign object distance GD2 is larger than the track distance GD1 and larger than the distance threshold value GD, and therefore it is determined that there is no centroid coordinate GD or less in the distance approaching difference image.

The track extraction unit 514 has a target track 1202a in the measurement image that is shorter than the preset distance threshold GD among the calculated track distances, and all the determined foreign object distances are distances. It determines with it being more than the use threshold value GD, and the target 1202a is extracted as a track of particles.

For example, in the measurement image 202e shown in FIG. 8C, the portion indicated by a solid line is extracted as a track of particles, and the portion indicated by a dotted line is excluded as a foreign matter or the like. The excluded portion 202f indicated by the dotted line at the center of the measurement image 202e is detected as a concave portion or a convex portion, as shown in FIGS. The foreign object distance is excluded because it is smaller than the distance threshold GD.

Next, the operation of the sample analyzer 100 of the present embodiment will be described with reference to FIGS.
As shown in FIG. 10, the sample analysis process starts when the sample 1 is supported on the upper surface of the sample table 22 and the imaging region of the sample 1 is set.
First, the arithmetic processing unit 51 moves the XY stage 21 to an area on the sample 1 where imaging is first started via the moving unit 2 (step S101).
Next, the arithmetic processing unit 51 detects the imaging surface by autofocus processing, and moves the focal position of the objective lens 31 to the reference position via the Z-axis control unit 36 (step S102).
Next, the arithmetic processing unit 51 moves the focal position of the objective lens 31 to the separation position via the Z-axis control unit 36, and images the separation image 201 by the imaging unit 123 (step S103).
Next, the arithmetic processing unit 51 moves the focal position of the objective lens 31 to the measurement position via the Z-axis control unit 36, and images the measurement image 202 by the imaging unit 123 (step S104).
Next, the arithmetic processing unit 51 moves the focal position of the objective lens 31 to the approach position via the Z-axis control unit 36, and captures the approach image 203 by the image capturing unit 123 (step S105).

Next, the arithmetic processing unit 51 performs the target image extraction process of the measurement image shown in FIG. 11 (step S106), which will be described later, and the target object extraction process of each difference image shown in FIG. 12, which will be described later (step S107). The track extraction process (step S108) shown in FIG. 13 is executed.
Next, the arithmetic processing unit 51 determines whether or not all areas have been imaged (step S109), and when not imaging (step S109; No), to the next area via the moving unit 2. The XY stage 21 is moved (step S110), and the processes of steps S102 to S108 are repeated.
And the arithmetic processing part 51 complete | finishes a sample analysis process, when imaging all the areas (step S109; Yes).

In the target image extraction process of the measurement image shown in FIG. 11, first, the target object extraction unit 512 performs a smoothing process (step S201) and a boundary extraction process on the captured measurement image 202 (step S202). The boundaries connected in units of pixels are labeled one by one (step S203).
Next, the target object extraction unit 512 determines whether or not a plurality of labeled boundaries are multiplexed (multiple circles) (step S204), and when they are multiplexed (step S204; Yes). Then, the boundary other than the boundary having the lowest average luminance value in the region within the boundary is removed (step S205), and step S204 is repeated.
Then, the target object extraction unit 512 ends the target object extraction process of the measurement image when there are no multiple boundaries or when all the multiple boundaries are removed (No in step S204).

In the target extraction processing of each difference image shown in FIG. 12, first, the difference acquisition unit 511 performs smoothing processing on the separation image 201 and the approach image 203 (step S301), and then approaches the separation image 201 and the separation approach. The difference image 205 is acquired (step S302).
Next, the target object extraction unit 512 extracts and labels the target object having the brightness value of the positive information in the difference images 204 and 205 (step S303).
Next, the target object extraction unit 512 removes target objects whose degree of coincidence with a track or a foreign object is equal to or less than a threshold value from among the labeled target objects (step S304).
Then, the target object extraction unit 512 acquires the barycentric coordinates of the remaining labeled target objects (step S305), and ends the target object extraction process of each difference image.

In the track extraction process shown in FIG. 13, the track extraction unit 514 first selects one target in the measurement image 202 having a labeled boundary (step S401).
Next, the non-extraction target removal unit 514b determines whether or not the boundary length is shorter than the boundary threshold LL (step S402). If the boundary length is shorter (step S402; Yes), the selected target is selected. It removes as a foreign material (step S410).
In addition, when the boundary length is equal to or greater than the boundary threshold LL (step S402; No), the non-extraction target removal unit 514b obtains an ellipse that approximates the contour of the object by the ellipse fitting process (step S403).
Next, the non-extraction target removal unit 514b determines whether or not the area of the ellipse obtained in step S403 is smaller than the area minimum value SL (step S404). If the area is smaller (step S404; Yes), the selection is performed. The completed target is removed as a foreign object (step S410).

Further, when the area of the ellipse is equal to or larger than the area minimum value SL (step S404; No), the non-extraction target removing unit 514b determines whether the area of the ellipse is larger than the area maximum value SH (step S404). S405).
When the area of the ellipse is larger than the maximum area value SH (step S405; Yes), the non-extraction target removal unit 514b removes the selected target as a foreign object (step S410). If the area of the ellipse is less than or equal to the area maximum value SH (step S405; No), it is determined whether or not the value obtained by dividing the minor axis of the ellipse by the major axis is smaller than the ellipse threshold OD (step S406).

If the divided value is smaller than the ellipse threshold OD (step S406; Yes), the non-extraction target removal unit 514b removes the selected target as a foreign object (step S410).
When the divided value is equal to or less than the ellipse threshold OD (step S406; No), the track extraction unit 514 determines whether or not the area of the ellipse is larger than the area threshold SE (step S407).
When the area of the ellipse is larger than the area threshold value SE (step S407; Yes), the track extraction unit 514 extracts the selected target as a track of particles (step S411).
When the area of the ellipse is equal to or smaller than the area threshold value SE (step S407; No), the track extraction unit 514 detects positive information in the approach / separation difference image 204 that is labeled with the barycentric coordinates of the target in the measurement image 202. The distance for the track, which is the distance from the center of gravity coordinates of the target having the brightness value, is obtained, and the foreign matter removing unit 513 includes the distance approach difference image 205 labeled with the center of gravity coordinates of the target in the measurement image 202. A distance for a foreign object which is a distance from the center of gravity coordinates of the target having the luminance value of the positive information is obtained (step S408).

Next, the track extraction unit 514 determines whether there is a track distance shorter than the distance threshold GD and there is no foreign object distance shorter than the distance threshold GD (step S409).
When there is no track distance shorter than the distance threshold GD, or when there is a distance for foreign matter shorter than the distance threshold GD (step S409; No), the foreign matter removing unit 513 selects the selected target. It removes as a foreign material (step S410).
When there is a track distance shorter than the distance threshold GD and there is no foreign object distance shorter than the distance threshold GD (step S409; Yes), the track extraction unit 514 is selected. The target is extracted as a track of particles (step S411).
Next, the track extraction unit 514 determines whether or not all the targets in the measurement image 202 have been selected (step S412). If not selected (step S412; No), the process proceeds to step S401. Return The next object is selected, and if it has been selected (step S412; Yes), the track extraction process is terminated.

As described above, according to the sample analyzer 100 of the present embodiment, by moving the focal position of the objective lens 31 in the Z-axis direction, the convex portion protruding from the imaging surface (the portion having the function of a convex lens) and , A concave portion (a portion having a function of a concave lens) recessed from the imaging surface can be detected. As a result, the sample analysis apparatus 100 of the present embodiment can automatically detect and remove the foreign matter 1a attached to the imaging surface when analyzing the track detection solid, and the track 1b of the particles recessed from the imaging surface. Only can be extracted.

Further, in the sample analyzer 100 of the present embodiment, only parallel light (collected parallel light) is emitted from the condenser lens 23 to the sample 1 to analyze the sample. The structure which radiates | emits to the sample 1 is unnecessary. As a result, the sample analyzer 100 of the present embodiment can omit the configuration (the diffuser plate and the second light source unit) for causing the collected diffused light to be incident on the film, and can be manufactured to detect convex portions and concave portions. Cost can be reduced.
In the sample analyzer 100 of the present embodiment, the separation position and the approach position are adjusted so that the contrast (for example, the difference value of the luminance value) between the convex portion and the concave portion and the other portion is increased. It is possible to accurately detect convex portions (for example, foreign matter 1a) and concave portions (for example, track 1b of particles).

In particular, in the sample analyzer 100 of the present embodiment, a target object having a positive luminance value is extracted from each of the difference images 204 and 205 acquired from the separated image 201 and the approach image 203, The contrast between the concave portion (the target having the luminance value of the positive information) and the other portion (the black portion other than the target) is further increased. As a result, the sample analyzer 100 according to the present embodiment can detect the convex portion (for example, the foreign matter 1a) and the concave portion (for example, the track 1b of the particles) with higher accuracy.
Further, in the sample analyzer 100 of the present embodiment, the target of the so-called just focus measurement image 202 is extracted and placed in the position of the target having a positive luminance value in each of the difference images 204 and 205. These are detected as convex portions and concave portions, and the convex portions and concave portions can be detected with higher accuracy.

Further, in the sample analyzer 100 of the present embodiment, when the boundary of the target in the measurement image 202 is multiple (multiple circle), the boundary other than the boundary having the lowest average luminance value in the boundary region is detected. Remove. As a result, the sample analyzer 100 according to the present embodiment can prevent erroneous detection in which the same convex portion or concave portion in the measurement image 202 is redundantly detected as a plurality of convex portions or concave portions.

Furthermore, in the sample analyzer 100 of this embodiment, in order to extract the track 1b of the particle formed as a circular recess, the track among the targets having the luminance value of the positive information in the difference images 204 and 205. Or those having a low degree of coincidence with a foreign object are removed, or those different from the particle track 1b are removed as non-extraction targets using the thresholds LL, SE, SL, SH, OD, and GD. As a result, the sample analysis apparatus 100 of the present embodiment can accurately remove the foreign matter 1a and the like in the measurement image 202 and extract only the particle tracks 1b with high accuracy.

As shown in FIG. 15, the control computer 150 of the sample analyzer 100 includes a control unit 151, a main storage unit 152, an external storage unit 153, an operation unit 154, a display unit 52, an input / output unit 156, and a transmission / reception unit 157. Prepare.
The main storage unit 152, the external storage unit 153, the operation unit 154, the display unit 52, the input / output unit 156, and the transmission / reception unit 157 are all connected to the control unit 151 via the internal bus 160.

The control unit 151 includes a CPU (Central Processing Unit) and the like, and according to a control program 158 stored in the external storage unit 153, a difference acquisition unit 511, a target extraction unit 512, a foreign matter removal unit 513 of the sample analyzer 100, and Each process of the track extraction unit 514 is executed. The arithmetic processing unit 51 is configured by the control unit 151.

The main storage unit 152 includes a RAM (Random-Access Memory) and the like, loads the control program 158 stored in the external storage unit 153, and functions as a work area of the control unit 151. The image RAM 124 is configured by the main storage unit 152.

The external storage unit 153 includes a non-volatile memory such as a flash memory, a hard disk, a DVD-RAM (Digital Versatile Disc Random-Access Memory), a DVD-RW (Digital Versatile Disc Disc ReWritable), and controls the processing of the sample analyzer 100. 151 stores a program to be executed in advance. Further, the external storage unit 153 supplies the stored data to the control unit 151 in accordance with an instruction from the control unit 151, and stores the data supplied from the control unit 151. The image storage device 53 is configured by an external storage unit 153.

The operation unit 154 includes a keyboard, a pointing device such as a mouse, and an interface device that connects the keyboard and the pointing device to the internal bus 160.

The display unit 52 includes a CRT (Cathode Ray Tube) or an LCD (Liquid Crystal Display), and displays the images 201 to 205, the operation screen of the sample analyzer 100, and the like.

The input / output unit 156 includes a serial interface or a parallel interface. The input / output unit 156 is connected to the microscope unit 110.

The transmission / reception unit 157 includes a network termination device or a wireless communication device connected to the network, and a serial interface or a LAN (Local Area Network) interface connected thereto.

The control program 158 performs processing using the control unit 151, the main storage unit 152, the external storage unit 153, the operation unit 154, the display unit 52, the input / output unit 156, the transmission / reception unit 157, and the like as resources, and is shown in FIG. The processing of the difference acquisition unit 511, the target object extraction unit 512, the foreign matter removal unit 513, and the track extraction unit 514 of the sample analyzer 100 is executed.

In the present embodiment, the sample 1 is composed of the track detection solid and the track 1b of the particles is extracted. For example, the sample 1 is a film described in Patent Documents 1 and 2, and the film is a film. It is good also as a structure which detects a crack (concave part) and dust (convex part).

In the present embodiment, the image sensor 4 is used as the imaging unit 123. However, for example, a line sensor or the like may be used.

In this embodiment, an example in which a CCD sensor is used as the image sensor 4 has been described. However, for example, a CMOS (Complementary Metal-Oxide Semiconductor) sensor or the like may be used.

In this embodiment, one separation position, one measurement position, and one approach position are set. However, for example, two or more can be set. In this case, the most in-focus measurement images are selected, the one with the highest brightness value of the target is selected from among the plurality of remote images, or the plurality of approach images are selected. It is possible to select the target having the maximum luminance value of the positive information and having the maximum luminance value.

In this embodiment, the boundary extraction process (edge extraction process) for extracting the boundary in the measurement image 202 is executed and the boundaries connected in units of pixels are labeled one by one. For example, as shown in FIG. Alternatively, the boundary of the target may be labeled after the measurement image 202 is converted into the monochrome image 202g by binarization extraction processing.

In the present embodiment, in the track extraction process, even a target that is extracted as a target having a positive brightness value in the approaching / separating difference image 204 is extracted as a target in the measurement image 202. If not, the structure is not extracted as the particle track 1b, but is not limited thereto. For example, when the target is extracted as a target having a positive luminance value in the approaching / separating difference image 204, the corresponding portion in the measurement image 202 is re-image processed (neighbor re-image processing). And it is good also as a structure which discriminate | determines whether it extracts as the track 1b of particle | grains.

In the present embodiment, in the track extraction process, the distance between the target in the selected measurement image 202 and the target having the luminance values of all the labeled positive information in the difference images 204 and 205. It is determined whether or not there is a track distance smaller than the distance threshold GD and there is no foreign object distance smaller than the distance threshold GD, but the present invention is not limited to this. For example, it may be determined whether or not the track distance is smaller than the distance threshold GD while obtaining the order of the label numbers. In this case, depending on the label number of the target in each of the difference images 204 and 205 determined to be separated from the target in the measurement image 202 by a track distance smaller than the distance threshold GD, the calculation is performed more than in the embodiment. The load can be reduced.

Other than the above, the above hardware configuration, flowchart, threshold value, parameter, etc. are merely examples, and can be arbitrarily changed and modified.

In the present embodiment, the case where parallel light is used for illumination has been described. However, the light may not necessarily be parallel light, and the visible light that can be extracted using the approaching / separating difference image and the approaching / separating difference image. Everything is targeted.

When each function of the sample analyzer 100 is realized by sharing of an OS (operating system) and an application program, or by cooperation between the OS and an application program (sample analysis program), only the application program portion is externally provided. You may store in the memory | storage part 153, a recording medium, a memory | storage device, etc.

It is also possible to superimpose an application program (sample analysis program) on a carrier wave and distribute it via a communication network. For example, an application program may be posted on a bulletin board (BBS: Bulletin Board System) on a communication network, and the application program may be distributed via the network. Then, the application program may be installed and activated in a computer, and may be configured to execute the above-described processing by being executed in the same manner as other application programs under the control of the OS.

The present invention is capable of various embodiments and modifications without departing from the broad spirit and scope of the present invention. Further, the above-described embodiment is for explaining the present invention, and does not limit the scope of the present invention. That is, the scope of the present invention is shown not by the embodiments but by the claims. Various modifications within the scope of the claims and within the scope of the equivalent invention are considered to be within the scope of the present invention.

This application is based on Japanese Patent Application No. 2012-206493 filed on September 20, 2012. The specification, claims, and entire drawings of Japanese Patent Application No. 2012-206493 are incorporated herein by reference.

DESCRIPTION OF SYMBOLS 1 ... Sample 1a ... Foreign substance 1b ... Track of particle 22 ... Sample stand 36 ... Z-axis control part 41 ... Output part 100 ... Sample analysis apparatus 123 ... Imaging part 201, 201a, 201c ... Separation image 202, 202a, 202e, 202g ... Measurement images 202b to 202d ... Boundaries 203, 203a, 203c ... Approaching images 204, 204a ... Approaching / separating difference image 205, 205a ... Separating approaching difference image 511 ... Difference acquiring unit 512 ... Target object extracting unit 513 ... Foreign object removing unit 513a ... Convex Detection unit 514 ... Track extraction unit 514a ... Concave detection unit

Claims (6)

1. An emission part for emitting visible light;
   A support part that is arranged on the optical axis of visible light emitted by the emission part and supports a sample through which visible light is transmitted;
   An imaging unit that has an optical system and captures an image of the sample supported by the support unit;
   The focal position of the optical system is set along the optical axis of visible light, the approach position closer to the emitting portion than the reference position of the imaging surface, which is the surface of the sample on the imaging portion side, and the emitting position than the reference position. A focal point moving part that is moved between a remote position away from the part,
   The focus moving unit moves the focus position to the approach position, and the brightness value of the approach image that is the image of the sample captured by the imaging unit at the approach position is larger than the brightness value of the peripheral portion that is set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position by the focus moving unit moving the focal position to the remote position, is smaller than the luminance value of the peripheral portion. Concave detecting means for detecting a portion as a concave recessed from the imaging surface;
   A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. Detection means;
   A sample analyzing apparatus comprising:
2. The approach / separation difference image obtained by subtracting the brightness value of each pixel corresponding to the separation image from the brightness value of each pixel of the approach image, and the brightness of each pixel corresponding to the approach image from the brightness value of each pixel of the separation image A difference acquisition means for acquiring a distance approaching difference image obtained by subtracting a value;
   A closed region extracting means for extracting a closed region whose luminance value is different from the peripheral portion from the image;
   Further comprising
   The closed region extracting means moves the focus position to a measurement position moved by a preset measurement distance from the reference position along the optical axis, and the imaging unit images at the measurement position. A closed region in the measurement image as the image of the sample, a closed region in which the luminance value in the approaching / separating difference image is a positive value, and a closed region in which the luminance value in the separating / approaching difference image is a positive value. Extract the region and
   The concave detecting means detects, as the concave portion, a closed region in the measurement image that is arranged at a position of the closed region where the brightness value in the approaching / separating difference image is a positive value,
   The convex detection means detects, as the convex portion, a closed region in the measurement image that is arranged at a position of a closed region where the brightness value in the separation approach difference image is a positive value. The sample analysis apparatus according to claim 1, characterized in that:
3. When the closed region extraction unit extracts an inner boundary and an outer boundary surrounding the inner boundary as a boundary surrounding a portion having a luminance value different from other portions from the measurement image, The sample analysis apparatus according to claim 2, wherein a boundary of a region having the lowest average luminance value is defined as a boundary of a closed region in the measurement image.
4. An imaging step of capturing an image of the sample by an imaging unit having an optical system when visible light is transmitted through the sample disposed on the optical axis;
   The focal position of the optical system is closer to the light source than the reference position of the imaging surface, which is the surface on the imaging unit side of the sample, along the optical axis of visible light, and away from the light source than the reference position A focal point moving step for moving between the separated position and the focal point moving unit;
   The focus moving unit moves the focus position to the approach position, and the brightness value of the approach image that is the image of the sample captured by the imaging unit at the approach position is larger than the brightness value of the peripheral portion that is set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position by the focus moving unit moving the focal position to the remote position, is smaller than the luminance value of the peripheral portion. A concave detecting step of detecting a portion as a concave recessed from the imaging surface;
   A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. A detection process;
   A sample analysis method comprising:
5. Computer
   An imaging unit that captures an image of the sample by an imaging unit having an optical system when visible light passes through the sample disposed on the optical axis;
   The focal position of the optical system is closer to the light source than the reference position of the imaging surface, which is the surface on the imaging unit side of the sample, along the optical axis of visible light, and away from the light source than the reference position A focal point moving means for moving between the separated position and
   The focus moving means moves the focus position to the approach position, and the brightness value of the approach image, which is the image of the sample imaged by the imaging unit at the approach position, is larger than the brightness value of the peripheral portion set in advance. In addition, the brightness value of the remote image, which is the image of the sample imaged by the imaging unit at the remote position with the focus moving means moving the focus position to the remote position, is smaller than the luminance value of the peripheral portion. Concave detecting means for detecting a portion as a concave recessed from the imaging surface;
   A convex portion that detects a portion where the luminance value of the separated image is larger than the luminance value of the peripheral portion and the luminance value of the approaching image is smaller than the luminance value of the peripheral portion as a convex portion protruding from the imaging surface. Detection means,
   Sample analysis program characterized by functioning as
6. An emission part for emitting visible light;
   A support part that is disposed on the optical axis of visible light emitted by the emission part and supports a track detection solid that transmits visible light; and
   An imaging unit that has an optical system and captures an image of a track detection solid as a sample supported by the support;
   The focal position of the optical system is closer to the emitting part than the reference position of the imaging surface, which is the surface on the imaging part side of the solid for track detection, along the optical axis of visible light, and from the reference position And a focal point moving unit that moves between a remote position away from the emitting unit,
   The brightness value of the peripheral portion in which the brightness value of the separated image, which is an image of the track detection solid image captured by the imaging unit at the separated position by the focus moving unit moving the focal position to the separated position, is set in advance. And the brightness value of the approach image, which is an image of the track detection solid image captured by the imaging unit at the approach position when the focus moving unit moves the focus position to the approach position, A foreign matter removing means for removing a portion smaller than the brightness value from the image of the track detection solid as a foreign matter attached to the imaging surface and protruding from the imaging surface;
   A track of particles formed by recessing a portion where the luminance value of the approaching image is larger than the luminance value of the peripheral portion and the luminance value of the separated image is smaller than the luminance value of the peripheral portion from the imaging surface Track extraction means for extracting from the track detection solid image as,
   A particle track analysis apparatus comprising:

Images