WO2015046518A1 - Sliced specimen evaluation device, sliced specimen preparation device including said sliced specimen evaluation device, and sliced specimen evaluation method - Google Patents

Sliced specimen evaluation device, sliced specimen preparation device including said sliced specimen evaluation device, and sliced specimen evaluation method Download PDF

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Publication number
WO2015046518A1
WO2015046518A1 PCT/JP2014/075914 JP2014075914W WO2015046518A1 WO 2015046518 A1 WO2015046518 A1 WO 2015046518A1 JP 2014075914 W JP2014075914 W JP 2014075914W WO 2015046518 A1 WO2015046518 A1 WO 2015046518A1
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WIPO (PCT)
Prior art keywords
sample
thin
section
unit
surface state
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PCT/JP2014/075914
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French (fr)
Japanese (ja)
Inventor
祐子 南條
健吾 渡辺
弘明 飯田
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倉敷紡績株式会社
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Priority to JP2015539430A priority Critical patent/JP6483022B2/en
Publication of WO2015046518A1 publication Critical patent/WO2015046518A1/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
    • G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G01N1/312—Apparatus therefor for samples mounted on planar substrates

Definitions

  • the present invention relates to a thin-section sample evaluation device for evaluating a thin-section sample used for physicochemical sample analysis or microscopic observation of a biological sample, a thin-section sample preparation device including the thin-section sample evaluation device, and a thin section
  • the present invention relates to a section sample evaluation method.
  • a microtome is widely known as an apparatus for producing a thin-section sample used for physicochemical sample analysis or microscopic observation of a biological sample.
  • the microtome is an apparatus for producing a thin slice sample by slicing a surface layer portion of a sample block in which a biological sample or the like is embedded (embedded) in an embedding agent such as paraffin by a cutter.
  • a thin slice sample prepared by a microtome is collected in a tank filled with a liquid for extension such as water or hot water using a brush or paper, and then stretched with a wrinkle or the like, and then an adhesive solution (for example, water ) To attach to the slide glass.
  • a liquid for extension such as water or hot water using a brush or paper
  • an adhesive solution for example, water
  • the said thin slice sample is directly arrange
  • the thin slice sample attached to the slide glass is tightly fixed to the slide glass as the adhesive solution evaporates, and is used as a thin slice sample for tissue observation (see, for example, Patent Documents 1-5).
  • the biological sample contained in the sample block may contain bone and calcified hard tissue.
  • the cutter When the surface of the sample block including such a hard tissue is sliced, the cutter may be damaged, and defects such as blade scratches may occur in the thin slice sample. Once the cutter is damaged, blade scratches are generated in all subsequent thin slice samples. Therefore, it is necessary to check whether there are any defects such as blade scratches on the surface of the thin slice sample.
  • An object of the present invention is to detect and evaluate a surface state of a thin slice sample obtained by slicing, a thin slice sample evaluation device, a thin slice sample preparation device including the thin slice sample evaluation device, and a thin slice sample preparation Is to provide a method.
  • the thin-section sample evaluation apparatus is a thin-section sample that evaluates the surface state of a thin-section sample placed on a slide glass by slicing a sample block in which the sample is embedded in an embedding agent
  • An evaluation device An irradiation unit for irradiating the surface of the thin-section sample placed on the slide glass with blue light
  • An imaging unit that receives diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit, and obtains image data of light of two or more different colors from the received light
  • a surface state detection unit that detects a surface state of the thin-section sample based on the image data about the light of two or more colors obtained by the imaging unit
  • a control unit that controls at least the imaging unit and the surface state detection unit, and evaluates the surface state of the thin-section sample detected by the surface state detection unit; Is provided.
  • the surface of the thin-section sample placed on the slide glass is irradiated with blue light, and diffusely reflected light from the surface of the thin-section sample is received to obtain two different colors.
  • the surface state of the thin slice sample is detected.
  • the surface condition of the detected thin slice sample is evaluated for the presence or absence of defects such as blade scratches, holes, fading, and placement failure.
  • FIG. 1 is a schematic diagram showing a configuration of a thin-section sample evaluation device according to Embodiment 1.
  • FIG. It is a top view of an example of the image of the thin section sample mounted on the slide glass.
  • It is a flowchart of the thin slice sample evaluation method which concerns on Embodiment 1 of this invention.
  • the thin-section sample evaluation apparatus slices a sample block in which a sample is embedded in an embedding agent with a cutter, and determines the surface state of the thin-section sample placed on a slide glass.
  • a thin section sample evaluation device for evaluation An irradiation unit for irradiating the surface of the thin-section sample placed on the slide glass with blue light;
  • An imaging unit that receives diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit, and obtains image data of light of two or more different colors from the received light;
  • a surface state detection unit that detects a surface state of the thin-section sample based on the image data about the light of two or more colors obtained by the imaging unit;
  • a control unit that controls at least the imaging unit and the surface state detection unit, and evaluates the surface state of the thin-section sample detected by the surface state detection unit; Is provided.
  • the thin-section sample evaluation apparatus is the thin-section sample evaluation apparatus according to the first aspect, wherein the irradiation unit is configured so that moisture exists between the thin-section sample and the slide glass.
  • the surface may be irradiated with blue light.
  • the thin-section sample evaluation apparatus is the first or second aspect, wherein the imaging unit is disposed vertically above the thin-section sample,
  • the said irradiation part may be arrange
  • the thin-section sample evaluation apparatus is the above-described third aspect, wherein the irradiation unit may be linear illumination.
  • the thin-section sample evaluation apparatus may be configured such that the irradiation unit is an annular irradiation unit arranged so as to surround the imaging unit.
  • the thin-section sample preparation apparatus is a thin-section sample preparation apparatus that prepares a thin-section sample by slicing a sample block in which a sample is embedded in an embedding agent with a cutter, A sample block transport unit for transporting the sample block; A cutter unit that slices the surface of the sample block with a cutter to obtain a thin slice sample; A placement section for placing the thin slice sample on a slide glass;
  • the thin-section sample evaluation device according to any one of the first to fifth aspects; At least a control unit that controls the sample block transport unit, the placement unit, and the thin-section sample evaluation device, Is provided.
  • a thin-section sample preparation device is the above-described sixth aspect, wherein the thin-section sample evaluation device detects the defect in the surface state of the thin-section sample when the thin section sample evaluation apparatus detects the cutter of the cutter section. It may be exchanged.
  • the thin-section sample preparation device stops the preparation of the thin-section sample in the sixth aspect when the thin-section sample evaluation device detects a defect in the surface state of the thin-section sample. May be.
  • the sample block in which the sample is embedded in the embedding agent is sliced with a cutter, and the surface state of the thin slice sample placed on the slide glass is measured.
  • a thin section sample evaluation method to be evaluated An irradiation step of irradiating the surface of the thin slice sample placed on the slide glass with blue light;
  • a surface state detection step of detecting a surface state of the thin-section sample based on the image data for the light of two or more colors obtained by the imaging unit;
  • An evaluation step for evaluating the surface condition of the detected thin slice sample including.
  • FIG. 1 is a schematic diagram showing an overall configuration of a thin-section sample preparation apparatus 100 according to Embodiment 1 of the present invention.
  • the thin-section sample preparation apparatus 100 mounts a thin section sample on a slide glass, a transport section 20 that transports the sample block 1, a cutter section 30 that slices the surface of the sample block 1 to obtain a thin section sample, and the like. It is only necessary to include at least the placement unit 60 to be placed, the thin-section sample evaluation device 10, and the control unit 90.
  • the control unit 90 controls the sample block transport unit 20, the cutter unit 30, the placement unit 60, and the thin-section sample evaluation device 10.
  • the control unit 90 may also serve as the control unit of the thin-section sample evaluation apparatus 10.
  • the thin-section sample preparation device 100 may further include a sample block storage unit 40, a position adjustment unit 50, an extension unit 70, and a slide glass conveyance unit 80. Below, each structural member which comprises the thin slice sample preparation apparatus 100 is demonstrated.
  • the sample block 1 is obtained by embedding (embedding) a biological sample 11 or the like in an embedding agent 12 such as paraffin.
  • the sample block 1 is used on a sample table (not shown).
  • a sample table not shown.
  • the sample block 1 When the sample block 1 is cut with the surface being dried, it is difficult to obtain a good section such as a sample having a fluffy cross section.
  • the sample block 1 has a problem that expansion and contraction are likely to occur due to a subtle change in temperature, and unevenness is likely to occur in the thickness of the thin slice sample during the thinning process.
  • a cooler and a humidifier are provided so as to maintain a constant temperature (for example, 10 ° C. to 25 ° C.) and a constant humidity.
  • the temperature and humidity may be set as appropriate according to the type of sample 11 and embedding agent 12 used as the sample block 1.
  • sample block storage unit The sample block storage unit 40 stores a plurality of sample blocks 1 in a state where they can be taken out.
  • the surface layer portion of the sample block is sliced by moving the cutter 31 and the sample block 1 relative to each other to obtain a thin slice sample 32.
  • the sample block 1 may be moved along the transport direction by the transport unit 20, and the cutter 31 may be moved in a direction perpendicular to the transport direction.
  • the cutter 31 instead of fixing it, the portion of the blade in contact with the sample block 1 can be shifted, and the paraffin waste generated during thin cutting can be easily removed.
  • the cutter unit 30 performs rough cutting for chamfering the sample block 1 and main cutting for producing a thin slice sample.
  • rough cutting the surface layer portion of the sample block 1 is cut to a predetermined thickness and surfaced.
  • main cutting the surface of the sample block 1 is sliced with a cutter 31 to produce a thin slice sample 32.
  • the prepared thin section sample 32 is attached on the carrier tape 34 supplied from the supply reel 33. In this case, the thin slice sample 32 can be satisfactorily adhered onto the carrier tape 34 by performing humidification.
  • the cutter unit 30 may also be provided with a height adjusting mechanism.
  • the carrier tape 34 is wound around the take-up reel 39 from the supply tape 33 through the guide rollers 35, 36, 37 and 38.
  • sample block transport unit 20 only needs to transport at least the sample block 1 to the cutter unit 30.
  • the placement unit 60 only needs to place at least the thin slice sample 32 on the upper surface of the slide glass 63.
  • the placement unit 60 includes a pair of guide rollers 61 disposed on the upstream side of the travel path of the carrier tape 34 and a pair of guide rollers 62 disposed on the downstream side of the travel path. It has.
  • a part of the carrier tape 21 to which the thin slice sample 32 is attached is sandwiched between the pair of guide rollers 61 and the pair of guide rollers 62, and in this state, the pair of guide rollers 62 or The carrier tape 21 is bent downward by moving the pair of guide rollers 61 downward.
  • the thin slice sample 32 is brought into contact with the upper surface of the slide glass 63 supplied with the adhesive liquid (including moisture) 64 on the upper surface, and the thin slice sample 32 is transferred to the upper surface of the slide glass 63.
  • the slide glass 63 on which the thin slice sample 32 is placed is called a slide glass with a thin slice sample.
  • the above-described transfer method is the most preferable.
  • a thin slice sample floated on the liquid in the liquid tank is placed on the slide glass. You may make it crawl up.
  • the extension unit 70 includes a heating plate (not shown).
  • the slide glass 63 with a thin-section sample placed on the heating plate (not shown) by the slide glass transport unit 80 is attached to the heating plate by the heating plate.
  • the first heating (for example, about 40 ° C. to 60 ° C., several seconds to several tens of seconds) is performed to extend the eyelids of the thin slice sample 32 and to increase the adhesive force of the thin slice sample 32 to the slide glass 63 .
  • the second heating for example, about 40 ° C. for several hours is further performed to completely evaporate the moisture on the slide glass 63 with the thin slice sample, and the thin slice sample 32 is fixedly adhered to the slide glass 63.
  • the thin-section sample preparation apparatus 100 includes the thin-section sample evaluation apparatus 10, the cutter section 30, the sample block transport section, and the placement section 60 as essential constituent members.
  • a sample block storage unit 40, a position adjustment unit 50, an extension unit 70, and a slide glass conveyance unit 80 may be provided.
  • FIG. 2 is a schematic diagram showing the configuration of the thin-section sample evaluation apparatus 10 according to Embodiment 1 of the present invention.
  • the thin-section sample evaluation device 10 includes irradiation units 3a and 3b, an imaging unit 4, a surface state detection unit 6, and a control unit 8.
  • the irradiation units 3a and 3b irradiate the surface of the thin slice sample 32 placed on the slide glass 63 with blue light.
  • the imaging unit 4 receives diffusely reflected light from the surface of the thin-section sample 32 of the light irradiated from the irradiation units 3a and 3b, and obtains image data for two or more different colors of light.
  • the surface state detection unit 6 detects the surface state of the thin slice sample 32 placed on the slide glass based on the image data regarding the light of two or more colors obtained by the imaging unit 4.
  • the control unit 8 controls the irradiation units 3 a and 3 b, the imaging unit 4, and the surface state detection unit 6.
  • blade scratches and holes Evaluate the presence or absence of defects such as faintness and poor placement.
  • the thin slice sample 32 In the evaluation of the thin slice sample 32, as shown in FIG. 1, when the thin slice sample 32 is placed on the slide glass 63, moisture 64 is supplied to the upper surface of the slide glass 63 in advance.
  • the thin section sample 32 may be placed on the surface.
  • the thin slice sample 32 By supplying moisture between the thin slice sample 32 and the slide glass 63 in this way, the thin slice sample 32 is brought into a wet state, so that the pattern of the sample (living tissue) 11 itself can be made difficult to see, and the blade scratches can be seen. , Holes, faintness, poor placement, etc. can be easily observed.
  • the effect of moisture existing between the thin slice sample 32 and the slide glass 63 will be described below.
  • the thin slice sample 32 includes a sample (living tissue) 11 and an embedding agent 12 such as paraffin.
  • scratches, holes, blurring, poor placement, etc. appearing on the surface of the thin slice sample 32 are similar to the pattern of the sample (biological tissue) 11 itself.
  • the surface state of the thin-section sample 32 is detected based on the image data of two or more different colors of light obtained by the imaging unit 4, and the surface of the obtained thin-section sample 32 is obtained. Regarding the state, it is possible to evaluate the presence or absence of defects such as blade scratches, holes, fading, and poor placement.
  • FIG. 3 is a plan view of an example of an image of the thin slice sample 32 placed on the slide glass 63.
  • This angle ⁇ can be detected, for example, by performing straight line extraction such as Hough transformation of image processing. It will be examined whether a series of scratches inclined at this angle ⁇ are blade scratches. The blade scratch is attached to the surface of the thin slice sample 32 when the sample is sliced with a damaged cutter.
  • the transport direction of the sample block 1 is from the left to the right of the page.
  • the blade scratches are inclined at an angle ⁇ with respect to the moving direction of the cutter.
  • This angle ⁇ corresponds to the apex angle ⁇ in the case of a right triangle having the moving speed of the cutter 31 and the transport speed of the sample block 1 as two components orthogonal to each other, and the transport speed component of the sample block 1 is the base.
  • the angle ⁇ of the series of scratches with respect to the moving direction of the cutter 31 is compared with the angle ⁇ with respect to the direction of movement of the cutter 31 in the case of blade scratches. This scratch is considered to be a blade scratch.
  • the cutter of the cutter unit 30 is replaced. Also good. Alternatively, the production of the thin slice sample 32 may be stopped.
  • the irradiation unit 3 (3a, 3b) can be used as long as it irradiates the surface of the sample block 1 with blue light.
  • Blue LED illumination may be used, or a blue color filter may be combined with white LED illumination or a white fluorescent lamp.
  • the peak wavelength of blue light is preferably in the range of 450 nm to 470 nm.
  • linear illumination may be used as the irradiation unit 3.
  • surface illumination may be used.
  • annular illumination unit in addition to the ring illumination, a plurality of dome illuminations or linear illuminations may be arranged in an annular shape to constitute the annular illumination unit.
  • the above description is an example and does not limit the range that can be used as the irradiation unit 3.
  • the imaging unit 4 can be used as long as it can obtain image data of light of two or more colors having different wavelengths.
  • a camera capable of acquiring a color image may be used.
  • an area camera or a line camera may be used.
  • the imaging unit 4 may be arranged in any way, but in the arrangement with the illuminating unit 3, the imaging unit 4 may be arranged so as to receive diffusely reflected light from the surface of the thin slice sample.
  • the imaging unit 4 may be arranged vertically above the thin slice sample. In this case, diffuse reflected light from the surface of the thin slice sample can be received unless the illumination unit 3 is arranged coaxially with the imaging unit 4.
  • the above description is an example and does not limit the range that can be used as the imaging unit 4.
  • the surface state detection unit 6 can be used as long as it has a function of detecting the surface state of the sliced piece sample based on the obtained image data of two or more colors of light.
  • color image data obtained from the imaging unit 4 may be processed to detect the surface state.
  • Image data processing methods include, for example, graying, shading correction, edge extraction, binarization, Hough conversion, and the like. Note that the image processing method is not limited to the above example.
  • the control unit 8 has a function of controlling at least the imaging unit 4 and the surface state detection unit 6 and evaluating the surface state based on the surface state obtained by the surface state detection unit 6. If you can use it.
  • the irradiation unit 3 may be further controlled by the control unit 8.
  • a personal computer can be used, for example.
  • a personal computer for example, a CPU, a ROM, a RAM, an HDD (storage unit), an input unit such as a touch panel, and a necessary member such as a display unit may be provided.
  • the surface state detection unit 6 and the control unit 8 are divided into two for convenience in order to distinguish them functionally, but may function as a physical configuration as a unit.
  • a program of a control unit operating on the CPU may control the surface state detection unit by calling the image processing program.
  • the image processing may be performed by a CPU built in the imaging unit 4. In that case, the imaging unit 4 and the surface state detection unit 6 are integrated as a physical configuration.
  • FIG. 4 is a flowchart of a method for evaluating a sliced piece sample in the method for preparing a sliced piece sample according to the first embodiment.
  • the thin slice sample evaluation method in this thin slice sample preparation method will be described below with reference to FIG. (A)
  • the thin slice sample 32 placed on the slide glass 63 is irradiated with blue light, diffusely reflected light from the surface of the thin slice sample 32 is received, and light of two or more different colors among the received light Image data is obtained (S01).
  • the image data obtained here may be color image data.
  • the reason for irradiating blue light is to improve the accuracy of detecting defects such as blade scratches and holes.
  • the diffusely reflected light is received because the surface of the thin-section sample 32 is irregularly reflected, and the surface of the slide glass 63 is dark without being irregularly reflected, so that the image data from the thin-section sample 32 is obtained. It is because it is easy to obtain.
  • the brightness of the slide glass 63 is higher than that of the thin slice sample 32 and the thin slice sample 32 becomes relatively dark, making it difficult to obtain image data suitable for evaluation. Become.
  • the surface state of the thin slice sample is detected (S02). For example, straight line flaws may be detected by using a green component in the obtained color image data to perform straight line extraction using Hough transform processing. Alternatively, binarization processing may be performed with a predetermined threshold value using a blue component of the obtained color image data, and a black pixel portion may be detected as a hole or a fading and a placement failure.
  • C The surface state of the obtained thin slice sample 32 is evaluated for defects such as blade scratches, holes, blurring, and placement failure (S03).
  • the angle of the linear flaw may be determined as a blade flaw by comparing it with the pulling angle of the cutter, or it may be determined as a defect if there is a hole having a certain area or more, a blur, or a placement failure.
  • the surface state of the thin-section sample 32 is measured based on the image data of two or more colors obtained by irradiating blue light. It can detect and evaluate defects such as misplacement. Therefore, by accurately evaluating the surface state of the thin slice sample 32 in the preparation of the thin slice sample, it is possible to detect the occurrence of blade flaws and suppress loss.
  • the said thin slice sample evaluation method can also be made into the computer program for implement
  • the computer program may be stored in a computer-readable recording medium.
  • the computer-readable recording medium may be, for example, an optical disk such as a CD-R, DVD, or BD, a semiconductor memory such as a USB memory or an SD memory, a magnetic disk, a magneto-optical disk, or a magnetic tape. .
  • FIG. 5 is a flowchart showing an outline of the thin-section sample preparation method according to Embodiment 1 of the present invention.
  • An outline of a method for producing the slide glass 63 with the thin-section sample 32 will be described with reference to FIG. (1) Thin cutting process The surface layer part of the sample block 1 is sliced with the cutter 31 of the cutter part 30, and the thin slice sample 32 is obtained (S11). (2) Placement process The thin slice sample 32 is placed on the slide glass 63 (S12). (3) Evaluation process The surface state of the thin-section sample 32 placed on the slide glass 63 is evaluated (S13). Specifically, it is determined whether or not the surface condition is good.
  • the surface state of the thin slice sample 32 is evaluated for the presence or absence of defects such as blade scratches, holes, fading, and poor placement. If there is a problem (NO), the process proceeds to step S14. If there is no problem, the process proceeds to step S15. (4) Replacing the cutter If the surface condition is not good, that is, if a defect such as a scratch on the blade, a hole, a blur, or a placement failure is found, the cutter 31 is replaced (S14). (5) Counting the number of slices It is determined whether the specified number of slices has been reached (S15). When the specified number of slices has been reached, the preparation of the sliced sample is finished (END). On the other hand, if the specified number of slicing has not been reached, the process returns to the first slicing process (S11). As described above, the thin slice sample 32 can be produced.
  • FIG. 6 is a flowchart showing details of a method for producing the slide glass 63 with the thin-section sample 32 according to Embodiment 1 of the present invention. Details of a method for producing the slide glass 63 with the thin-section sample 32 will be described below with reference to FIG.
  • B) The position adjustment unit 50 adjusts the inclination and height of the surface of the sample block 1 (S22).
  • the slide glass conveyance unit 80 conveys one slide glass 63 for attaching the thin slice sample 32 to the placement unit 60, After supplying the adhesive liquid 23 (including moisture) to the upper surface, the thin slice sample 32 and the slide glass 63 attached on the carrier tape 34 are brought into a facing state (S26).
  • the mounting unit 60 a part of the carrier tape 34 to which the thin slice sample 32 is attached is bent downward, and the thin slice sample 32 is pressed against the adhesive liquid 64 on the upper surface of the slide glass 63, thereby thinly The section sample 32 is transferred from a part of the carrier tape 34 to the upper surface of the slide glass 63 (S27).
  • the extension section 70 performs the first heating (for example, about 40 ° C. to 60 ° C., several seconds to several tens of seconds) on the conveyed slide glass 63 with the thin slice sample, and the thin slice sample 32. And the sticking force of the thin slice sample 32 to the slide glass 63 is strengthened. Thereafter, the second heating (for example, about 40 ° C. for several hours) is further performed to completely evaporate the moisture and to fix the thin slice sample 32 to the slide glass 22 (S30). Thereby, the production of the slide glass 63 with the thin slice sample 32 is completed. Thereafter, the sample block 1 is transported by the sample block transport unit 20 in the same manner as described above, and the thin slice operation is automatically and continuously repeated any number of times, thereby preparing an arbitrary number of thin slice samples 32. it can.
  • the first heating for example, about 40 ° C. to 60 ° C., several seconds to several tens of seconds
  • the thin-section sample is irradiated with blue light, and diffusely reflected light from the surface of the thin-section sample is received and received.
  • Image data is obtained for two or more different colors of light. Based on the obtained image data of two or more colors of light, the surface state of the thin slice sample can be detected and evaluated. Therefore, the present invention is useful for a thin-section sample preparation apparatus for preparing a thin-section sample used for physicochemical sample analysis or microscopic observation of biological samples.

Abstract

A sliced specimen evaluation device for using a cutter to slice a specimen block in which a specimen is embedded in an embedding medium, place the sliced specimen on an adhesive liquid supplied above slide glass, and evaluate the surface state of the sliced specimen placed on the adhesive liquid, said sliced specimen evaluation device being provided with an irradiation unit for irradiating blue light onto the surface of the sliced specimen placed on the adhesive liquid supplied above the slide glass, an imaging unit for receiving light that has been irradiated by the irradiation unit and diffusely reflected by the surface of the sliced specimen and obtaining image data for light of at least two different colors from among the received light, a surface state detection unit for detecting the surface state of the sliced specimen on the basis of the image data for the light of at least two colors obtained by the imaging unit, and a control unit for controlling at least the imaging unit and the surface state detection unit and evaluating the detected surface state of the sliced specimen.

Description

薄切片試料評価装置、該薄切片試料評価装置を含む薄切片試料作製装置、及び、薄切片試料評価方法Thin section sample evaluation apparatus, thin section sample preparation apparatus including the thin section sample evaluation apparatus, and thin section sample evaluation method
　本発明は、理化学試料分析や生体試料等の顕微鏡観察等に利用される薄切片試料を評価するための薄切片試料評価装置、該薄切片試料評価装置を含む薄切片試料作製装置、及び、薄切片試料評価方法に関する。 The present invention relates to a thin-section sample evaluation device for evaluating a thin-section sample used for physicochemical sample analysis or microscopic observation of a biological sample, a thin-section sample preparation device including the thin-section sample evaluation device, and a thin section The present invention relates to a section sample evaluation method.
　従来、理化学試料分析や生体試料等の顕微鏡観察等に利用される薄切片試料を作製するための装置としてミクロトームが広く知られている。ミクロトームは、生体試料等をパラフィン等の包埋剤の中に埋め込んだ（包埋した）試料ブロックの表層部分をカッターによって薄切りすることにより、薄切片試料を作製する装置である。 Conventionally, a microtome is widely known as an apparatus for producing a thin-section sample used for physicochemical sample analysis or microscopic observation of a biological sample. The microtome is an apparatus for producing a thin slice sample by slicing a surface layer portion of a sample block in which a biological sample or the like is embedded (embedded) in an embedding agent such as paraffin by a cutter.
　ミクロトームにより作製された薄切片試料は、例えば、筆や紙等を用いて、水あるいは湯等の伸展用の液体を充填した槽に回収されて皺等を伸展されたのち、接着液（例えば水）を用いてスライドガラスに貼付けられる。あるいは、上記薄切片試料は、接着剤を塗布したスライドガラスに直接配置され、スライドガラスが加温されることで皺等の伸展が行われてスライドガラスに貼付けられる。スライドガラスに貼り付いた薄切片試料は、接着液の蒸発に伴いスライドガラスに密着固定され、組織観察用の薄切片試料として利用される（例えば、特許文献１－５参照。）。 A thin slice sample prepared by a microtome is collected in a tank filled with a liquid for extension such as water or hot water using a brush or paper, and then stretched with a wrinkle or the like, and then an adhesive solution (for example, water ) To attach to the slide glass. Or the said thin slice sample is directly arrange | positioned on the slide glass which apply | coated the adhesive agent, and extension | extension of wrinkles etc. is performed and affixed on a slide glass by heating a slide glass. The thin slice sample attached to the slide glass is tightly fixed to the slide glass as the adhesive solution evaporates, and is used as a thin slice sample for tissue observation (see, for example, Patent Documents 1-5).
特開２０１２－２２９９９３号公報JP 2012-229993A 特開２０１２－２２９９９４号公報JP 2012-229994 A 特開２０１２－２２９９９５号公報JP 2012-229995 A 特開２０１２－２２９９９６号公報JP 2012-229996 A 特開２０１２－２２９９９７号公報JP 2012-229997 A
　試料ブロックに含まれる生体試料の中には骨および石灰化した硬組織等が含まれることがある。このような硬組織を含む試料ブロックの表面を薄切りした場合、カッターが損傷して薄切片試料に刃キズ等の不具合が生じる場合がある。いったんカッターが損傷すると、その後の全ての薄切片試料に刃キズを生じるため、薄切片試料の表面に刃キズ等の不具合が存在しないか確認することが必要である。 The biological sample contained in the sample block may contain bone and calcified hard tissue. When the surface of the sample block including such a hard tissue is sliced, the cutter may be damaged, and defects such as blade scratches may occur in the thin slice sample. Once the cutter is damaged, blade scratches are generated in all subsequent thin slice samples. Therefore, it is necessary to check whether there are any defects such as blade scratches on the surface of the thin slice sample.
　本発明の目的は、薄切りして得られた薄切片試料の表面状態を検出、評価する薄切片試料評価装置及び該薄切片試料評価装置を備えた薄切片試料作製装置、並びに、薄切片試料作製方法を提供することである。 An object of the present invention is to detect and evaluate a surface state of a thin slice sample obtained by slicing, a thin slice sample evaluation device, a thin slice sample preparation device including the thin slice sample evaluation device, and a thin slice sample preparation Is to provide a method.
　本発明に係る薄切片試料評価装置は、試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りし、スライドガラスの上に載置した薄切片試料の表面状態を評価する薄切片試料評価装置であって、
　前記スライドガラスの上に載置した前記薄切片試料の表面に青色光を照射する照射部と、
　前記照射部から照射した光の前記薄切片試料の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る撮像部と、
　前記撮像部で得られた前記２色以上の光についての前記画像データに基づいて、前記薄切片試料の表面状態を検出する表面状態検出部と、
　少なくとも、前記撮像部と、前記表面状態検出部と、を制御し、前記表面状態検出部で検出した前記薄切片試料の表面状態について評価する制御部と、
を備える。 The thin-section sample evaluation apparatus according to the present invention is a thin-section sample that evaluates the surface state of a thin-section sample placed on a slide glass by slicing a sample block in which the sample is embedded in an embedding agent An evaluation device,
An irradiation unit for irradiating the surface of the thin-section sample placed on the slide glass with blue light;
An imaging unit that receives diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit, and obtains image data of light of two or more different colors from the received light;
A surface state detection unit that detects a surface state of the thin-section sample based on the image data about the light of two or more colors obtained by the imaging unit;
A control unit that controls at least the imaging unit and the surface state detection unit, and evaluates the surface state of the thin-section sample detected by the surface state detection unit;
Is provided.
　本発明に係る薄切片試料評価装置によれば、スライドガラスの上に載置された薄切片試料の表面に青色光を照射し、薄切片試料の表面による拡散反射光を受光して異なる２色以上の光について得られた画像データに基づいて、薄切片試料の表面状態を検出する。検出した薄切片試料の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合の有無を評価する。これによって、薄切片試料の表面に刃キズ等の不具合が発生していた場合に、これを検出でき、その後の多数の薄切片のロスを抑制できると共に、薄切片試料の品質低下を抑制できる。 According to the thin-section sample evaluation apparatus according to the present invention, the surface of the thin-section sample placed on the slide glass is irradiated with blue light, and diffusely reflected light from the surface of the thin-section sample is received to obtain two different colors. Based on the image data obtained for the above light, the surface state of the thin slice sample is detected. The surface condition of the detected thin slice sample is evaluated for the presence or absence of defects such as blade scratches, holes, fading, and placement failure. As a result, when a defect such as a blade flaw has occurred on the surface of the thin slice sample, this can be detected, the loss of many subsequent thin slices can be suppressed, and the quality deterioration of the thin slice sample can be suppressed.
本発明の実施の形態１に係る薄切片試料作製装置の構成を示す概略図である。It is the schematic which shows the structure of the thin section sample preparation apparatus which concerns on Embodiment 1 of this invention. 実施の形態１に係る薄切片試料評価装置の構成を示す概略図である。1 is a schematic diagram showing a configuration of a thin-section sample evaluation device according to Embodiment 1. FIG. スライドガラスの上に載置した薄切片試料の画像の一例の平面図である。It is a top view of an example of the image of the thin section sample mounted on the slide glass. 本発明の実施の形態１に係る薄切片試料評価方法のフローチャートである。It is a flowchart of the thin slice sample evaluation method which concerns on Embodiment 1 of this invention. 本発明の実施の形態１に係る薄切片試料作製方法のフローチャートである。It is a flowchart of the thin slice sample preparation method which concerns on Embodiment 1 of this invention. 本発明の実施の形態１に係る薄切片試料付きスライドガラスの作製方法の詳細を示すフローチャートである。It is a flowchart which shows the detail of the manufacturing method of the slide glass with a thin section sample which concerns on Embodiment 1 of this invention.
　本発明の第１の態様に係る薄切片試料評価装置は、試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りし、スライドガラスの上に載置した薄切片試料の表面状態を評価する薄切片試料評価装置であって、
　前記スライドガラスの上に載置した前記薄切片試料の表面に青色光を照射する照射部と、
　前記照射部から照射した光の前記薄切片試料の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る撮像部と、
　前記撮像部で得られた前記２色以上の光についての前記画像データに基づいて、前記薄切片試料の表面状態を検出する表面状態検出部と、
　少なくとも、前記撮像部と、前記表面状態検出部と、を制御し、前記表面状態検出部で検出した前記薄切片試料の表面状態について評価する制御部と、
を備える。 The thin-section sample evaluation apparatus according to the first aspect of the present invention slices a sample block in which a sample is embedded in an embedding agent with a cutter, and determines the surface state of the thin-section sample placed on a slide glass. A thin section sample evaluation device for evaluation,
An irradiation unit for irradiating the surface of the thin-section sample placed on the slide glass with blue light;
An imaging unit that receives diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit, and obtains image data of light of two or more different colors from the received light;
A surface state detection unit that detects a surface state of the thin-section sample based on the image data about the light of two or more colors obtained by the imaging unit;
A control unit that controls at least the imaging unit and the surface state detection unit, and evaluates the surface state of the thin-section sample detected by the surface state detection unit;
Is provided.
　第２の態様に係る薄切片試料評価装置は、上記第１の態様において、前記照射部は、前記薄切片試料と、前記スライドガラスとの間に水分が存在する状態で、前記薄切片試料の表面に青色光を照射してもよい。 The thin-section sample evaluation apparatus according to a second aspect is the thin-section sample evaluation apparatus according to the first aspect, wherein the irradiation unit is configured so that moisture exists between the thin-section sample and the slide glass. The surface may be irradiated with blue light.
　第３の態様に係る薄切片試料評価装置は、上記第１又は第２の態様において、前記撮像部は、前記薄切片試料の鉛直上方に配置され、
　前記照射部は、前記撮像部を挟んで左右に配置されていてもよい。 The thin-section sample evaluation apparatus according to a third aspect is the first or second aspect, wherein the imaging unit is disposed vertically above the thin-section sample,
The said irradiation part may be arrange | positioned on either side on both sides of the said imaging part.
　第４の態様に係る薄切片試料評価装置は、上記第３の態様において、前記照射部は、直線照明であってもよい。 The thin-section sample evaluation apparatus according to the fourth aspect is the above-described third aspect, wherein the irradiation unit may be linear illumination.
　第５の態様に係る薄切片試料評価装置は、上記第３の態様において、前記照射部は、前記撮像部を囲むように配置された環状照射部であってもよい。 In the third aspect, the thin-section sample evaluation apparatus according to the fifth aspect may be configured such that the irradiation unit is an annular irradiation unit arranged so as to surround the imaging unit.
　本発明の第６の態様に係る薄切片試料作製装置は、試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りして薄切片試料を作製する薄切片試料作製装置であって、
　前記試料ブロックを搬送する試料ブロック搬送部と、
　前記試料ブロックの表面をカッターによって薄切りして薄切片試料を得るカッター部と、
　前記薄切片試料を、スライドガラスの上に載置する載置部と、
　上記第１から第５のいずれかの態様の前記薄切片試料評価装置と、
　少なくとも、前記試料ブロック搬送部と、前記載置部と、前記薄切片試料評価装置と、を制御する制御部と、
を備える。 The thin-section sample preparation apparatus according to the sixth aspect of the present invention is a thin-section sample preparation apparatus that prepares a thin-section sample by slicing a sample block in which a sample is embedded in an embedding agent with a cutter,
A sample block transport unit for transporting the sample block;
A cutter unit that slices the surface of the sample block with a cutter to obtain a thin slice sample;
A placement section for placing the thin slice sample on a slide glass;
The thin-section sample evaluation device according to any one of the first to fifth aspects;
At least a control unit that controls the sample block transport unit, the placement unit, and the thin-section sample evaluation device,
Is provided.
　第７の態様に係る薄切片試料作製装置は、上記第６の態様において、前記薄切片試料評価装置によって、前記薄切片試料の表面状態に不具合を検出した場合に、前記カッター部の前記カッターを交換してもよい。 A thin-section sample preparation device according to a seventh aspect is the above-described sixth aspect, wherein the thin-section sample evaluation device detects the defect in the surface state of the thin-section sample when the thin section sample evaluation apparatus detects the cutter of the cutter section. It may be exchanged.
　第８の態様に係る薄切片試料作製装置は、上記第６の態様において、前記薄切片試料評価装置によって、前記薄切片試料の表面状態に不具合を検出した場合に、薄切片試料の作製を停止してもよい。 The thin-section sample preparation device according to an eighth aspect stops the preparation of the thin-section sample in the sixth aspect when the thin-section sample evaluation device detects a defect in the surface state of the thin-section sample. May be.
　本発明の第９の態様に係る薄切片試料評価方法は、試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りし、スライドガラスの上に載置した薄切片試料の表面状態を評価する薄切片試料評価方法であって、
　前記スライドガラスの上に載置した前記薄切片試料の表面に青色光を照射する照射ステップと、
　前記照射部から照射した光の前記薄切片試料の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る撮像ステップと、
　前記撮像部で得られた前記２色以上の光についての前記画像データに基づいて、前記薄切片試料の表面状態を検出する表面状態検出ステップと、
　前記検出した前記薄切片試料の表面状態について評価する評価ステップと、
を含む。 In the thin slice sample evaluation method according to the ninth aspect of the present invention, the sample block in which the sample is embedded in the embedding agent is sliced with a cutter, and the surface state of the thin slice sample placed on the slide glass is measured. A thin section sample evaluation method to be evaluated,
An irradiation step of irradiating the surface of the thin slice sample placed on the slide glass with blue light;
An imaging step of receiving diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit and obtaining image data of light of two or more different colors among the received light;
A surface state detection step of detecting a surface state of the thin-section sample based on the image data for the light of two or more colors obtained by the imaging unit;
An evaluation step for evaluating the surface condition of the detected thin slice sample;
including.
　以下、本発明の実施の形態に係る薄切片試料評価装置、薄切片試料作製装置及び薄切片試料評価方法について、添付図面を参照しながら説明する。なお、図面において実質的に同一の部材については同一の符号を付している。 Hereinafter, a thin-section sample evaluation device, a thin-section sample preparation device, and a thin-section sample evaluation method according to embodiments of the present invention will be described with reference to the accompanying drawings. In the drawings, substantially the same members are denoted by the same reference numerals.
（実施の形態１）
＜薄切片試料作製装置＞
　図１は、本発明の実施の形態１に係る薄切片試料作製装置１００の全体的な構成を示す概略図である。この薄切片試料作製装置１００は、試料ブロック１を搬送する搬送部２０と、試料ブロック１の表面を薄切りして薄切片試料を得るカッター部３０と、薄切片試料を、スライドガラスの上に載置する載置部６０と、薄切片試料評価装置１０と、制御部９０と、を少なくとも備えていればよい。制御部９０は、試料ブロック搬送部２０と、カッター部３０と、載置部６０と、薄切片試料評価装置１０と、を制御する。なお、制御部９０は、薄切片試料評価装置１０の制御部を兼ねてもよい。 (Embodiment 1)
<Thin section sample preparation device>
FIG. 1 is a schematic diagram showing an overall configuration of a thin-section sample preparation apparatus 100 according to Embodiment 1 of the present invention. The thin-section sample preparation apparatus 100 mounts a thin section sample on a slide glass, a transport section 20 that transports the sample block 1, a cutter section 30 that slices the surface of the sample block 1 to obtain a thin section sample, and the like. It is only necessary to include at least the placement unit 60 to be placed, the thin-section sample evaluation device 10, and the control unit 90. The control unit 90 controls the sample block transport unit 20, the cutter unit 30, the placement unit 60, and the thin-section sample evaluation device 10. The control unit 90 may also serve as the control unit of the thin-section sample evaluation apparatus 10.
　この薄切片試料作製装置１００は、さらに、試料ブロック保管部４０と、位置調整部５０と、伸展部７０と、スライドガラス搬送部８０と、を備えていてもよい。
　以下に薄切片試料作製装置１００を構成する各構成部材について説明する。 The thin-section sample preparation device 100 may further include a sample block storage unit 40, a position adjustment unit 50, an extension unit 70, and a slide glass conveyance unit 80.
Below, each structural member which comprises the thin slice sample preparation apparatus 100 is demonstrated.
＜試料ブロック＞
　試料ブロック１は、例えば、生体試料１１等をパラフィン等の包埋剤１２の中に包埋（埋め込み）したものである。この試料ブロック１は、試料台（図示せず）の上に載せて使用される。この試料ブロック１は、表面が乾燥した状態で切断されると、試料断面が毛羽立つ等、良好な切片が得にくくなる。また、試料ブロック１は、温度の微妙な変化によって膨張及び収縮が発生しやすく、薄切り処理時に薄切片試料の厚さにムラが生じやすいという問題点を有する。このため、一定温度（例えば１０℃～２５℃）及び一定湿度を維持するように、図示しない冷却機及び加湿器を備えている。なお、上記温度及び湿度の設定は、試料ブロック１として使用する試料１１及び包埋剤１２の種類等に応じて適宜設定すればよい。 <Sample block>
The sample block 1 is obtained by embedding (embedding) a biological sample 11 or the like in an embedding agent 12 such as paraffin. The sample block 1 is used on a sample table (not shown). When the sample block 1 is cut with the surface being dried, it is difficult to obtain a good section such as a sample having a fluffy cross section. In addition, the sample block 1 has a problem that expansion and contraction are likely to occur due to a subtle change in temperature, and unevenness is likely to occur in the thickness of the thin slice sample during the thinning process. For this reason, a cooler and a humidifier (not shown) are provided so as to maintain a constant temperature (for example, 10 ° C. to 25 ° C.) and a constant humidity. The temperature and humidity may be set as appropriate according to the type of sample 11 and embedding agent 12 used as the sample block 1.
＜試料ブロック保管部＞
　試料ブロック保管部４０では、複数の試料ブロック１を取り出し可能な状態で保管する。
＜位置調整部＞
　位置調整部５０では、試料ブロック１の表面の傾斜及び高さ調整を行う。 <Sample block storage unit>
The sample block storage unit 40 stores a plurality of sample blocks 1 in a state where they can be taken out.
<Position adjustment unit>
The position adjustment unit 50 adjusts the inclination and height of the surface of the sample block 1.
＜カッター部＞
　カッター部３０では、カッター３１と試料ブロック１とを相対移動させることによって試料ブロックの表層部分を薄切りして薄切片試料３２を得る。この場合に、例えば、試料ブロック１は、搬送部２０によって搬送方向に沿って移動させ、カッター３１は、搬送方向と垂直方向に移動させてもよい。カッター３１を固定するのではなく移動させることによって、試料ブロック１と接する刃の部分をずらすことができ、薄切りの際に発生するパラフィンくずがとれやすくなる。 <Cutter part>
In the cutter unit 30, the surface layer portion of the sample block is sliced by moving the cutter 31 and the sample block 1 relative to each other to obtain a thin slice sample 32. In this case, for example, the sample block 1 may be moved along the transport direction by the transport unit 20, and the cutter 31 may be moved in a direction perpendicular to the transport direction. By moving the cutter 31 instead of fixing it, the portion of the blade in contact with the sample block 1 can be shifted, and the paraffin waste generated during thin cutting can be easily removed.
　また、カッター部３０では、試料ブロック１の面出しのための粗削りと、薄切片試料を作製する本削りと、を行う。
　粗削りの場合には、所定厚さまで試料ブロック１の表層部分を削って面出しを行う。
　一方、本削りの場合には、試料ブロック１の表面をカッター３１によって薄切りして薄切片試料３２を作製する。作製した薄切片試料３２は、供給リール３３から供給されるキャリアテープ３４の上に付着される。この場合に、加湿を行うことによって薄切片試料３２をキャリアテープ３４上に良好に付着することができる。なお、カッター部３０にも高さ調整機構を設けても良い。 The cutter unit 30 performs rough cutting for chamfering the sample block 1 and main cutting for producing a thin slice sample.
In the case of rough cutting, the surface layer portion of the sample block 1 is cut to a predetermined thickness and surfaced.
On the other hand, in the case of main cutting, the surface of the sample block 1 is sliced with a cutter 31 to produce a thin slice sample 32. The prepared thin section sample 32 is attached on the carrier tape 34 supplied from the supply reel 33. In this case, the thin slice sample 32 can be satisfactorily adhered onto the carrier tape 34 by performing humidification. The cutter unit 30 may also be provided with a height adjusting mechanism.
＜キャリアテープ＞
　なお、キャリアテープ３４は、供給テープ３３から、ガイドローラ３５、３６、３７、３８を経て、巻取リール３９に巻き取られる。 <Carrier tape>
The carrier tape 34 is wound around the take-up reel 39 from the supply tape 33 through the guide rollers 35, 36, 37 and 38.
＜試料ブロック搬送部＞
　試料ブロック搬送部２０は、少なくとも試料ブロック１をカッター部３０に搬送するものであればよい。 <Sample block transport section>
The sample block transport unit 20 only needs to transport at least the sample block 1 to the cutter unit 30.
＜載置部＞
　載置部６０は、少なくとも薄切片試料３２をスライドガラス６３の上面に載置するものであればよい。例えば、転写方式を用いた場合の載置部６０は、キャリアテープ３４の走行経路の上流側に配置された一対のガイドローラ６１と、走行経路の下流側に配置された一対のガイドローラ６２とを備えている。載置部６０では、例えば、一対のガイドローラ６１の間と一対のガイドローラ６２の間で薄切片試料３２が貼り付いているキャリアテープ２１の一部分を挟み、その状態で一対のガイドローラ６２又は一対のガイドローラ６１を下方に移動させることで、キャリアテープ２１を下方に撓ませる。これによって、上面に接着液（水分を含む）６４を供給されたスライドガラス６３の上面に薄切片試料３２を接触させて、スライドガラス６３の上面に薄切片試料３２を転写する。なお、薄切片試料３２を載置されたスライドガラス６３を、薄切片試料付きスライドガラスという。なお、薄切片試料３２をスライドガラス６３の上面に載置する方法としては前記転写による方法が最も好ましいが、その他の例としては、液槽内の液体に浮かべた薄切片試料を、スライドガラスで掬い上げるようにしてもよい。 <Placement part>
The placement unit 60 only needs to place at least the thin slice sample 32 on the upper surface of the slide glass 63. For example, when the transfer method is used, the placement unit 60 includes a pair of guide rollers 61 disposed on the upstream side of the travel path of the carrier tape 34 and a pair of guide rollers 62 disposed on the downstream side of the travel path. It has. In the mounting portion 60, for example, a part of the carrier tape 21 to which the thin slice sample 32 is attached is sandwiched between the pair of guide rollers 61 and the pair of guide rollers 62, and in this state, the pair of guide rollers 62 or The carrier tape 21 is bent downward by moving the pair of guide rollers 61 downward. Thus, the thin slice sample 32 is brought into contact with the upper surface of the slide glass 63 supplied with the adhesive liquid (including moisture) 64 on the upper surface, and the thin slice sample 32 is transferred to the upper surface of the slide glass 63. The slide glass 63 on which the thin slice sample 32 is placed is called a slide glass with a thin slice sample. As a method for placing the thin slice sample 32 on the upper surface of the slide glass 63, the above-described transfer method is the most preferable. However, as another example, a thin slice sample floated on the liquid in the liquid tank is placed on the slide glass. You may make it crawl up.
＜伸展部＞
　伸展部７０は、加温板（図示せず）を備え、スライドガラス搬送部８０により加温板（図示せず）上に載置された薄切片試料付きスライドガラス６３に、上記加温板により、第１の加温（例えば４０℃～６０℃程度、数秒～数十秒）を行って薄切片試料３２の皺の伸展を行うとともに薄切片試料３２のスライドガラス６３への貼付力を強くする。その後、さらに第２の加温（例えば４０℃程度、数時間）を行って薄切片試料付きスライドガラス６３上の水分を完全に蒸発させて、薄切片試料３２をスライドガラス６３に密着固定する。 <Extension Department>
The extension unit 70 includes a heating plate (not shown). The slide glass 63 with a thin-section sample placed on the heating plate (not shown) by the slide glass transport unit 80 is attached to the heating plate by the heating plate. The first heating (for example, about 40 ° C. to 60 ° C., several seconds to several tens of seconds) is performed to extend the eyelids of the thin slice sample 32 and to increase the adhesive force of the thin slice sample 32 to the slide glass 63 . Thereafter, the second heating (for example, about 40 ° C. for several hours) is further performed to completely evaporate the moisture on the slide glass 63 with the thin slice sample, and the thin slice sample 32 is fixedly adhered to the slide glass 63.
　なお、上述のように、この薄切片試料作製装置１００では、上記薄切片試料評価装置１０、カッター部３０、試料ブロック搬送部、載置部６０を必須の構成部材として含むが、これらの構成部材以外に、試料ブロック保管部４０と、位置調整部５０と、伸展部７０と、スライドガラス搬送部８０と、を備えていてもよい。 As described above, the thin-section sample preparation apparatus 100 includes the thin-section sample evaluation apparatus 10, the cutter section 30, the sample block transport section, and the placement section 60 as essential constituent members. In addition, a sample block storage unit 40, a position adjustment unit 50, an extension unit 70, and a slide glass conveyance unit 80 may be provided.
＜薄切片試料評価装置＞
　図２は、本発明の実施の形態１に係る薄切片試料評価装置１０の構成を示す概略図である。この薄切片試料評価装置１０は、照射部３ａ、３ｂと、撮像部４と、表面状態検出部６と、制御部８と、を備える。照射部３ａ、３ｂは、スライドガラス６３の上に載置した薄切片試料３２の表面に青色光を照射する。撮像部４は、照射部３ａ、３ｂから照射した光の薄切片試料３２の表面による拡散反射光を受光して、異なる２色以上の光についての画像データを得る。表面状態検出部６は、撮像部４で得られた２色以上の光についての画像データに基づいて、スライドガラスの上に載置した薄切片試料３２の表面状態を検出する。制御部８は、照射部３ａ、３ｂと、撮像部４と、表面状態検出部６と、を制御し、表面状態検出部６で得られた薄切片試料３２の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合の有無を評価する。 <Thin section sample evaluation device>
FIG. 2 is a schematic diagram showing the configuration of the thin-section sample evaluation apparatus 10 according to Embodiment 1 of the present invention. The thin-section sample evaluation device 10 includes irradiation units 3a and 3b, an imaging unit 4, a surface state detection unit 6, and a control unit 8. The irradiation units 3a and 3b irradiate the surface of the thin slice sample 32 placed on the slide glass 63 with blue light. The imaging unit 4 receives diffusely reflected light from the surface of the thin-section sample 32 of the light irradiated from the irradiation units 3a and 3b, and obtains image data for two or more different colors of light. The surface state detection unit 6 detects the surface state of the thin slice sample 32 placed on the slide glass based on the image data regarding the light of two or more colors obtained by the imaging unit 4. The control unit 8 controls the irradiation units 3 a and 3 b, the imaging unit 4, and the surface state detection unit 6. About the surface state of the thin slice sample 32 obtained by the surface state detection unit 6, blade scratches and holes Evaluate the presence or absence of defects such as faintness and poor placement.
　なお、薄切片試料３２の評価にあたって、図１に示すように、薄切片試料３２をスライドガラス６３の上に載置する際にあらかじめスライドガラス６３の上面に水分６４を供給しておき、その上に薄切片試料３２を載置してもよい。 In the evaluation of the thin slice sample 32, as shown in FIG. 1, when the thin slice sample 32 is placed on the slide glass 63, moisture 64 is supplied to the upper surface of the slide glass 63 in advance. The thin section sample 32 may be placed on the surface.
　このように薄切片試料３２とスライドガラス６３の間に水分を供給することによって、薄切片試料３２を濡れた状態とすることで、試料（生体組織）１１自体の模様を見えにくくでき、刃キズ、穴、かすれ、載置不良等の観察がしやすくなる。
　上記薄切片試料３２とスライドガラス６３の間に存在する水分の効果について以下に説明する。まず、薄切片試料３２には、試料（生体組織）１１とパラフィン等の包埋剤１２が含まれている。一方、薄切片試料３２の表面に現れる刃キズ、穴、かすれ、載置不良等は、試料（生体組織）１１自体の模様と類似する。そのため、試料（生体組織）１１を乾燥した状態においた場合、試料（生体組織）１１自体の模様が表面に浮き出てしまい、表面の刃キズ、穴、かすれ、載置不良等がまぎれてしまい観察できなくなるおそれがある。これに対して、薄切片試料３２とスライドガラス６３の間に水分が存在していると、試料（生体組織）１１とパラフィンとの輝度差がつきにくくなる。このように試料（生体組織）１１とパラフィンとの輝度差があまりないことは、試料（生体組織）１１自体の観察を行う場合には不利となるが、ここでは表面状態を検出、評価することが目的であるので上記輝度差がないことは特に問題とならない。その一方、薄切片試料３２とスライドガラス６３の間に水分が存在することによって、試料（生体組織）１１自体の模様を見えにくくできるため、表面の刃キズ、穴、かすれ、載置不良等の観察がしやすくなる。 By supplying moisture between the thin slice sample 32 and the slide glass 63 in this way, the thin slice sample 32 is brought into a wet state, so that the pattern of the sample (living tissue) 11 itself can be made difficult to see, and the blade scratches can be seen. , Holes, faintness, poor placement, etc. can be easily observed.
The effect of moisture existing between the thin slice sample 32 and the slide glass 63 will be described below. First, the thin slice sample 32 includes a sample (living tissue) 11 and an embedding agent 12 such as paraffin. On the other hand, scratches, holes, blurring, poor placement, etc. appearing on the surface of the thin slice sample 32 are similar to the pattern of the sample (biological tissue) 11 itself. Therefore, when the sample (biological tissue) 11 is in a dry state, the pattern of the sample (biological tissue) 11 itself is raised on the surface, and the surface blade scratches, holes, blurring, mounting defects, etc. are covered and observed. There is a risk that it will not be possible. On the other hand, if moisture exists between the thin-section sample 32 and the slide glass 63, a difference in luminance between the sample (living tissue) 11 and paraffin is less likely to occur. Thus, the fact that there is not much difference in luminance between the sample (living tissue) 11 and paraffin is disadvantageous when observing the sample (living tissue) 11 itself, but here the surface state is detected and evaluated. Therefore, the absence of the luminance difference is not particularly problematic. On the other hand, the presence of moisture between the thin-section sample 32 and the slide glass 63 makes it difficult to see the pattern of the sample (living tissue) 11 itself, so that surface blade scratches, holes, blurring, poor placement, etc. Easy to observe.
　この薄切片試料評価装置１０では、撮像部４で得られた異なる２色以上の光についての画像データに基づいて、薄切片試料３２の表面状態を検出し、得られた薄切片試料３２の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合の有無を評価することができる。 In the thin-section sample evaluation device 10, the surface state of the thin-section sample 32 is detected based on the image data of two or more different colors of light obtained by the imaging unit 4, and the surface of the obtained thin-section sample 32 is obtained. Regarding the state, it is possible to evaluate the presence or absence of defects such as blade scratches, holes, fading, and poor placement.
　図３は、スライドガラス６３の上に載置した薄切片試料３２の画像の一例の平面図である。この画像では、図３で矢印で示すカッター３１の移動方向（紙面の上から下の方向）に対して角度βで傾斜して生じている一連のキズがあることがわかる。この角度βは、例えば、画像処理のハフ（Ｈｏｕｇｈ）変換等の直線抽出を行うことによって検出できる。この角度βで傾斜している一連のキズが刃キズであるかについて検討する。刃キズとは、損傷したカッターによって試料を薄切りする際に、薄切片試料３２の表面に付けられるものである。なお、図３の場合、試料ブロック１の搬送方向は、紙面の左から右の方向である。上記のようにカッター部３０において、試料ブロック１の搬送方向と、カッター３１の移動方向とが互いに直交している場合、刃キズはカッターの移動方向について角度αで傾斜して生じる。この角度αは、カッター３１の移動速度と試料ブロック１の搬送速度とを互いに直交する２つの成分とする直角三角形において、試料ブロック１の搬送速度の成分を底辺とした場合の頂角αに対応する。そこで、上記一連のキズのカッター３１の移動方向に対する角度βについて、刃キズの場合のカッター３１の移動方向に対する角度αと対比し、角度βが角度αと実質的に一致する場合には上記一連のキズは刃キズであると考えられる。 FIG. 3 is a plan view of an example of an image of the thin slice sample 32 placed on the slide glass 63. In this image, it can be seen that there are a series of flaws that occur at an angle β with respect to the moving direction of the cutter 31 indicated by the arrow in FIG. 3 (from the top to the bottom of the paper). This angle β can be detected, for example, by performing straight line extraction such as Hough transformation of image processing. It will be examined whether a series of scratches inclined at this angle β are blade scratches. The blade scratch is attached to the surface of the thin slice sample 32 when the sample is sliced with a damaged cutter. In the case of FIG. 3, the transport direction of the sample block 1 is from the left to the right of the page. As described above, in the cutter unit 30, when the transport direction of the sample block 1 and the moving direction of the cutter 31 are orthogonal to each other, the blade scratches are inclined at an angle α with respect to the moving direction of the cutter. This angle α corresponds to the apex angle α in the case of a right triangle having the moving speed of the cutter 31 and the transport speed of the sample block 1 as two components orthogonal to each other, and the transport speed component of the sample block 1 is the base. To do. Therefore, the angle β of the series of scratches with respect to the moving direction of the cutter 31 is compared with the angle α with respect to the direction of movement of the cutter 31 in the case of blade scratches. This scratch is considered to be a blade scratch.
　この薄切片試料作製装置１００では、薄切片試料３２の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合が存在すると評価された場合には、カッター部３０のカッターを交換してもよい。あるいは、薄切片試料３２の作製を停止してもよい。 In the thin-section sample preparation device 100, if it is evaluated that the surface state of the thin-section sample 32 has a defect such as a flaw, a hole, a blur, or a placement failure, the cutter of the cutter unit 30 is replaced. Also good. Alternatively, the production of the thin slice sample 32 may be stopped.
　この薄切片試料評価装置１０を構成する各構成部材について説明する。 Each component constituting the thin slice sample evaluation apparatus 10 will be described.
　＜照射部＞
　照射部３（３ａ、３ｂ）は、試料ブロック１の表面に青色光を照射するものであれば使用できる。青色ＬＥＤ照明を用いてもよいし、白色ＬＥＤ照明や白色の蛍光ランプに青色のカラーフィルターを組み合わせてもよい。なお、青色光のピーク波長は４５０ｎｍ－４７０ｎｍの範囲にあるのが好ましい。さらに、照射部３としては、直線照明を用いてもよい。あるいは、面照明を用いてもよい。なお、照射部３として直線照明を用いる場合には、左右に直線照明３ａ、３ｂを配置してもよい。さらに、撮像部４を囲むように環状照明部（図示せず）としてもよい。環状照明部としては、リング状照明の他、ドーム照明または、直線照明を複数環状に並べて環状照明部を構成してもよい。
　なお、上記記載は一例であって照射部３として利用可能な範囲を限定するものではない。 <Irradiation part>
The irradiation unit 3 (3a, 3b) can be used as long as it irradiates the surface of the sample block 1 with blue light. Blue LED illumination may be used, or a blue color filter may be combined with white LED illumination or a white fluorescent lamp. Note that the peak wavelength of blue light is preferably in the range of 450 nm to 470 nm. Further, as the irradiation unit 3, linear illumination may be used. Alternatively, surface illumination may be used. In addition, when using linear illumination as the irradiation part 3, you may arrange | position the linear illumination 3a, 3b on either side. Furthermore, it is good also as an annular illumination part (not shown) so that the imaging part 4 may be enclosed. As the annular illumination unit, in addition to the ring illumination, a plurality of dome illuminations or linear illuminations may be arranged in an annular shape to constitute the annular illumination unit.
The above description is an example and does not limit the range that can be used as the irradiation unit 3.
　＜撮像部＞
　撮像部４は、異なる波長の２色以上の光についての画像データを得ることができるものであれば使用できる。例えば、カラー画像が取得できるカメラであってもよい。また、撮像部４として、エリアカメラを用いても良く、あるいはラインカメラを用いてもよい。
　また、撮像部４は、どのように配置してもよいが、照明部３との配置において、薄切片試料の表面からの拡散反射光を受光できるように配置すればよい。例えば、撮像部４は、薄切片試料の鉛直上方に配置してもよい。この場合、照明部３を撮像部４と同軸に配置しない限り薄切片試料の表面からの拡散反射光を受光することができる。なお、上記記載は一例であって撮像部４として利用可能な範囲を限定するものではない。 <Imaging unit>
The imaging unit 4 can be used as long as it can obtain image data of light of two or more colors having different wavelengths. For example, a camera capable of acquiring a color image may be used. Further, as the imaging unit 4, an area camera or a line camera may be used.
In addition, the imaging unit 4 may be arranged in any way, but in the arrangement with the illuminating unit 3, the imaging unit 4 may be arranged so as to receive diffusely reflected light from the surface of the thin slice sample. For example, the imaging unit 4 may be arranged vertically above the thin slice sample. In this case, diffuse reflected light from the surface of the thin slice sample can be received unless the illumination unit 3 is arranged coaxially with the imaging unit 4. The above description is an example and does not limit the range that can be used as the imaging unit 4.
　＜表面状態検出部及び制御部＞
　表面状態検出部６は、得られた２色以上の光についての画像データに基づいて、薄切片試料の表面状態を検出する機能を有するものであれば使用できる。例えば、撮像部４から得たカラー画像データを処理し、表面状態を検出してもよい。画像データの処理方法としては、例えば、グレー化、シェーディング補正、エッジ抽出、２値化、ハフ（Ｈｏｕｇｈ）変換、等がある。なお、画像処理の方法は、上記の例に限定されない。また、制御部８は、少なくとも、撮像部４と、表面状態検出部６と、を制御し、表面状態検出部６で得られた表面状態に基づいて、表面状態を評価する機能を有するものであれば使用できる。なお、制御部８によって、さらに照射部３を制御してもよい。また、表面状態検出部６及び制御部８としては、例えば、パーソナルコンピュータを使用できる。パーソナルコンピュータとしては、例えば、ＣＰＵ、ＲＯＭ、ＲＡＭ、ＨＤＤ（記憶部）、タッチパネル等の入力部、表示部等の必要な部材を備えていればよい。
　なお、表面状態検出部６及び制御部８は、機能的に区別するため便宜的に２つに分けているが、物理的な構成として一体として機能するものであってもよい。例えば、ＣＰＵ上で動作する制御部のプログラムが、上記画像処理のプログラムを呼び出すことで表面状態検出部を制御してもよい。また、撮像部４に内蔵されたＣＰＵで上記画像処理を行ってもよい。その場合、撮像部４と表面状態検出部６は物理的な構成として一体となる。 <Surface condition detection unit and control unit>
The surface state detection unit 6 can be used as long as it has a function of detecting the surface state of the sliced piece sample based on the obtained image data of two or more colors of light. For example, color image data obtained from the imaging unit 4 may be processed to detect the surface state. Image data processing methods include, for example, graying, shading correction, edge extraction, binarization, Hough conversion, and the like. Note that the image processing method is not limited to the above example. The control unit 8 has a function of controlling at least the imaging unit 4 and the surface state detection unit 6 and evaluating the surface state based on the surface state obtained by the surface state detection unit 6. If you can use it. The irradiation unit 3 may be further controlled by the control unit 8. Moreover, as the surface state detection part 6 and the control part 8, a personal computer can be used, for example. As a personal computer, for example, a CPU, a ROM, a RAM, an HDD (storage unit), an input unit such as a touch panel, and a necessary member such as a display unit may be provided.
Note that the surface state detection unit 6 and the control unit 8 are divided into two for convenience in order to distinguish them functionally, but may function as a physical configuration as a unit. For example, a program of a control unit operating on the CPU may control the surface state detection unit by calling the image processing program. Further, the image processing may be performed by a CPU built in the imaging unit 4. In that case, the imaging unit 4 and the surface state detection unit 6 are integrated as a physical configuration.
＜薄切片試料の表面状態の評価方法＞
　図４は、実施の形態１に係る薄切片試料作製方法における、薄切片試料の評価方法のフローチャートである。この薄切片試料作製方法における、薄切片試料評価方法について、図４を用いて以下に説明する。
（ａ）スライドガラス６３の上に載置した薄切片試料３２に青色光を照射し、薄切片試料３２の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る（Ｓ０１）。ここで得られる画像データは、カラー画像データであってもよい。また、青色光を照射するのは、刃キズや穴といった不具合の検出精度を高めるためである。青色光を照射し、その反射光の緑色成分で刃キズおよびシワを、青色成分で穴、かすれ、載置不良を判別することで、それぞれの不具合を高精度に検出できる。なお、拡散反射光を受光するのは、この場合に薄切片試料３２の表面が乱反射するため明るく、一方、スライドガラス６３の表面は乱反射せず暗くなるため、薄切片試料３２からの画像データを得やすいためである。これに対して、正反射光を受光した場合には、薄切片試料３２よりもスライドガラス６３の輝度が高くなって薄切片試料３２が相対的に暗くなり、評価に適した画像データを得にくくなる。
（ｂ）得られた２色以上の光についての画像データに基づいて、薄切片試料の表面状態を検出する（Ｓ０２）。例えば、得られたカラー画像データのうち緑色成分を使って、ハフ変換処理を用いた直線抽出を行い直線状のキズを検出してもよい。また、得られたカラー画像データのうちの青色成分を使って所定の閾値で２値化処理を行い、黒画素の部分を穴、かすれ、載置不良として検出してもよい。
（ｃ）得られた薄切片試料３２の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合を評価する（Ｓ０３）。例えば、直線状のキズの角度をカッターの引き角と比較して刃キズと判定してもよいし、一定以上の面積の穴、かすれ、載置不良があれば不具合と判定してもよい。 <Method for evaluating surface condition of thin slice sample>
FIG. 4 is a flowchart of a method for evaluating a sliced piece sample in the method for preparing a sliced piece sample according to the first embodiment. The thin slice sample evaluation method in this thin slice sample preparation method will be described below with reference to FIG.
(A) The thin slice sample 32 placed on the slide glass 63 is irradiated with blue light, diffusely reflected light from the surface of the thin slice sample 32 is received, and light of two or more different colors among the received light Image data is obtained (S01). The image data obtained here may be color image data. Moreover, the reason for irradiating blue light is to improve the accuracy of detecting defects such as blade scratches and holes. By irradiating blue light and discriminating blade scratches and wrinkles with the green component of the reflected light, and holes, fading and mounting defects with the blue component, it is possible to detect each defect with high accuracy. In this case, the diffusely reflected light is received because the surface of the thin-section sample 32 is irregularly reflected, and the surface of the slide glass 63 is dark without being irregularly reflected, so that the image data from the thin-section sample 32 is obtained. It is because it is easy to obtain. On the other hand, when regular reflection light is received, the brightness of the slide glass 63 is higher than that of the thin slice sample 32 and the thin slice sample 32 becomes relatively dark, making it difficult to obtain image data suitable for evaluation. Become.
(B) Based on the obtained image data of two or more colors of light, the surface state of the thin slice sample is detected (S02). For example, straight line flaws may be detected by using a green component in the obtained color image data to perform straight line extraction using Hough transform processing. Alternatively, binarization processing may be performed with a predetermined threshold value using a blue component of the obtained color image data, and a black pixel portion may be detected as a hole or a fading and a placement failure.
(C) The surface state of the obtained thin slice sample 32 is evaluated for defects such as blade scratches, holes, blurring, and placement failure (S03). For example, the angle of the linear flaw may be determined as a blade flaw by comparing it with the pulling angle of the cutter, or it may be determined as a defect if there is a hole having a certain area or more, a blur, or a placement failure.
　この薄切片試料評価方法では、青色光を照射し、得られた２色以上の光についての画像データに基づいて、薄切片試料３２の表面状態、具体的には刃キズ、穴、かすれ、載置不良等の不具合を検出、評価できる。そこで、薄切片試料作製にあたって薄切片試料３２の表面状態の評価を的確に行うことによって、刃キズ等の発生を検出し、ロスを抑えることができる。 In this thin-section sample evaluation method, the surface state of the thin-section sample 32, specifically, scratches, holes, blurs, and the like, is measured based on the image data of two or more colors obtained by irradiating blue light. It can detect and evaluate defects such as misplacement. Therefore, by accurately evaluating the surface state of the thin slice sample 32 in the preparation of the thin slice sample, it is possible to detect the occurrence of blade flaws and suppress loss.
　なお、上記薄切片試料評価方法は、コンピュータ上で実行することによって、コンピュータによる薄切片試料評価方法を実現するためのコンピュータプログラムとすることもできる。この場合、上記コンピュータプログラムは、コンピュータ読み取り可能な記録媒体に格納してもよい。また、コンピュータ読み取り可能な記録媒体としては、例えば、ＣＤ－Ｒ、ＤＶＤ、ＢＤ等の光ディスク、ＵＳＢメモリ、ＳＤメモリ等の半導体メモリ、磁気ディスク、光磁気ディスク等、磁気テープ等であってもよい。 In addition, the said thin slice sample evaluation method can also be made into the computer program for implement | achieving the thin slice sample evaluation method by a computer by performing on a computer. In this case, the computer program may be stored in a computer-readable recording medium. The computer-readable recording medium may be, for example, an optical disk such as a CD-R, DVD, or BD, a semiconductor memory such as a USB memory or an SD memory, a magnetic disk, a magneto-optical disk, or a magnetic tape. .
＜薄切片試料作製方法＞
　次に、図５は、本発明の実施の形態１に係る薄切片試料作製方法の概略を示すフローチャートである。図５を用いて、薄切片試料３２付きスライドガラス６３の作製方法の概略を説明する。
（１）薄切処理
　試料ブロック１の表層部分をカッター部３０のカッター３１によって薄切りして薄切片試料３２を得る（Ｓ１１）。
（２）載置処理
　薄切片試料３２をスライドガラス６３の上に載置する（Ｓ１２）。
（３）評価処理
　スライドガラス６３の上に載置した薄切片試料３２の表面状態を評価する（Ｓ１３）。具体的には、表面状態が良好か否か判断する。この場合、薄切片試料３２の表面状態について、刃キズ、穴、かすれ、載置不良等の不具合の有無を評価する。
　不具合があった場合（ＮＯ）には、ステップＳ１４に移行する。不具合がなかった場合には、そのままステップＳ１５に移行する。
（４）カッターの交換
　表面状態がよくない場合、つまり、刃キズ、穴、かすれ、載置不良等の不具合が見つかった場合にはカッター３１の交換を行う（Ｓ１４）。
（５）薄切回数のカウント
　規定の薄切回数に達したか判断する（Ｓ１５）。規定の薄切回数に達した場合には薄切片試料の作製を終了する（エンド）。一方、規定の薄切回数に達していない場合には、最初の薄切処理（Ｓ１１）に戻る。
　以上によって、薄切片試料３２の作製を行うことができる。 <Thin section sample preparation method>
Next, FIG. 5 is a flowchart showing an outline of the thin-section sample preparation method according to Embodiment 1 of the present invention. An outline of a method for producing the slide glass 63 with the thin-section sample 32 will be described with reference to FIG.
(1) Thin cutting process The surface layer part of the sample block 1 is sliced with the cutter 31 of the cutter part 30, and the thin slice sample 32 is obtained (S11).
(2) Placement process The thin slice sample 32 is placed on the slide glass 63 (S12).
(3) Evaluation process The surface state of the thin-section sample 32 placed on the slide glass 63 is evaluated (S13). Specifically, it is determined whether or not the surface condition is good. In this case, the surface state of the thin slice sample 32 is evaluated for the presence or absence of defects such as blade scratches, holes, fading, and poor placement.
If there is a problem (NO), the process proceeds to step S14. If there is no problem, the process proceeds to step S15.
(4) Replacing the cutter If the surface condition is not good, that is, if a defect such as a scratch on the blade, a hole, a blur, or a placement failure is found, the cutter 31 is replaced (S14).
(5) Counting the number of slices It is determined whether the specified number of slices has been reached (S15). When the specified number of slices has been reached, the preparation of the sliced sample is finished (END). On the other hand, if the specified number of slicing has not been reached, the process returns to the first slicing process (S11).
As described above, the thin slice sample 32 can be produced.
＜薄切片試料３２付きスライドガラス６３の作製方法＞
　図６は、本発明の実施の形態１に係る薄切片試料３２付きスライドガラス６３の作製方法の詳細を示すフローチャートである。図６を用いて、薄切片試料３２付きスライドガラス６３の作製方法の詳細について、以下に説明する。
（ａ）まず、試料保管部４０から一つの試料ブロック１を試料ブロック搬送部２０により取り出し、位置調整部５０に搬送する（Ｓ２１）。
（ｂ）位置調整部５０において、試料ブロック１の表面の傾斜及び高さ調整を行う（Ｓ２２）。
（ｃ）次いで、試料ブロック１を、試料ブロック搬送部２０によって、位置調整部５０からカッター部３０に搬送する（Ｓ２３）。
（ｄ）カッター部３０では、試料ブロック搬送部２０とカッター３１との相対移動によって、試料ブロック１の表層部分をカッター３１によって薄切する（Ｓ２４）。これによって、一枚の薄切片試料３２が作製され、薄切片試料３２がキャリアテープ３４に貼り付けられる。
（ｅ）キャリアテープ３４に貼り付けられた薄切片試料３２を、キャリアテープ３４の移動と共に、載置部６０に移動させる（Ｓ２５）。
（ｆ）一方、試料ブロック１の移動及び薄切片試料３２の移動の間に、スライドガラス搬送部８０によって、薄切片試料３２を貼り付ける１つのスライドガラス６３を載置部６０に搬送して、上面に接着液２３（水分を含む）を供給した後、キャリアテープ３４上に貼り付けられた薄切片試料３２とスライドガラス６３とを対向状態とする（Ｓ２６）。
（ｇ）載置部６０では、薄切片試料３２が貼り付けられたキャリアテープ３４の一部分を下方に撓ませて、薄切片試料３２をスライドガラス６３の上面の接着液６４に押し当てて、薄切片試料３２をキャリアテープ３４の一部分からスライドガラス６３の上面に転写する（Ｓ２７）。
（ｈ）薄切片試料評価装置１０によって、スライドガラス６３の上に載置された薄切片試料３２の表面状態を検出、評価する（Ｓ２８）。この薄切片試料評価方法については、上述の通りであるため説明を省略する。
（ｉ）薄切片試料３２が載置された薄切片試料付きスライドガラス６３をスライドガラス搬送部８０により、載置部６０から伸展部７０へ搬送する（Ｓ２９）。 <Method for Producing Slide Glass 63 with Thin Section Sample 32>
FIG. 6 is a flowchart showing details of a method for producing the slide glass 63 with the thin-section sample 32 according to Embodiment 1 of the present invention. Details of a method for producing the slide glass 63 with the thin-section sample 32 will be described below with reference to FIG.
(A) First, one sample block 1 is taken out from the sample storage unit 40 by the sample block transport unit 20 and transported to the position adjustment unit 50 (S21).
(B) The position adjustment unit 50 adjusts the inclination and height of the surface of the sample block 1 (S22).
(C) Next, the sample block 1 is conveyed from the position adjusting unit 50 to the cutter unit 30 by the sample block conveying unit 20 (S23).
(D) In the cutter unit 30, the surface layer portion of the sample block 1 is sliced by the cutter 31 by the relative movement of the sample block transport unit 20 and the cutter 31 (S24). As a result, one thin slice sample 32 is produced, and the thin slice sample 32 is attached to the carrier tape 34.
(E) The thin slice sample 32 attached to the carrier tape 34 is moved to the placement unit 60 along with the movement of the carrier tape 34 (S25).
(F) On the other hand, during the movement of the sample block 1 and the movement of the thin slice sample 32, the slide glass conveyance unit 80 conveys one slide glass 63 for attaching the thin slice sample 32 to the placement unit 60, After supplying the adhesive liquid 23 (including moisture) to the upper surface, the thin slice sample 32 and the slide glass 63 attached on the carrier tape 34 are brought into a facing state (S26).
(G) In the mounting unit 60, a part of the carrier tape 34 to which the thin slice sample 32 is attached is bent downward, and the thin slice sample 32 is pressed against the adhesive liquid 64 on the upper surface of the slide glass 63, thereby thinly The section sample 32 is transferred from a part of the carrier tape 34 to the upper surface of the slide glass 63 (S27).
(H) The surface state of the thin slice sample 32 placed on the slide glass 63 is detected and evaluated by the thin slice sample evaluation device 10 (S28). Since this thin slice sample evaluation method is as described above, the description thereof is omitted.
(I) The slide glass 63 with the thin section sample on which the thin section sample 32 is placed is transported from the placement section 60 to the extension section 70 by the slide glass transport section 80 (S29).
（ｋ）伸展部７０では、搬送された薄切片試料付きスライドガラス６３に対して、第１の加温（例えば４０℃～６０℃程度、数秒～数十秒）を行って、薄切片試料３２の皺の伸展を行うとともに薄切片試料３２のスライドガラス６３への貼付力を強化する。その後、さらに第２の加温（例えば４０℃程度、数時間）を行って、水分を完全に蒸発させて薄切片試料３２をスライドガラス２２に密着固定させる（Ｓ３０）。
　これにより、一枚の薄切片試料３２付きスライドガラス６３の作製が完了する。
　以下、上記と同様にして試料ブロック搬送部２０により試料ブロック１を搬送して、上記薄切り動作を、任意の回数、自動的且つ連続的に繰り返すことによって、任意の枚数の薄切片試料３２を作製できる。 (K) The extension section 70 performs the first heating (for example, about 40 ° C. to 60 ° C., several seconds to several tens of seconds) on the conveyed slide glass 63 with the thin slice sample, and the thin slice sample 32. And the sticking force of the thin slice sample 32 to the slide glass 63 is strengthened. Thereafter, the second heating (for example, about 40 ° C. for several hours) is further performed to completely evaporate the moisture and to fix the thin slice sample 32 to the slide glass 22 (S30).
Thereby, the production of the slide glass 63 with the thin slice sample 32 is completed.
Thereafter, the sample block 1 is transported by the sample block transport unit 20 in the same manner as described above, and the thin slice operation is automatically and continuously repeated any number of times, thereby preparing an arbitrary number of thin slice samples 32. it can.
　なお、本発明は、上記実施の形態に限定されるものではなく、その他種々の態様で実施できる。 Note that the present invention is not limited to the above embodiment, and can be implemented in various other modes.
　本発明に係る薄切片試料評価装置及び該評価装置を含む薄切片試料作製装置によれば、青色光を薄切片試料に照射し、薄切片試料の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る。得られた２色以上の光についての画像データに基づいて、薄切片試料の表面状態を検出、評価できる。そこで、理化学試料分析や生体試料等の顕微鏡観察などに利用される薄切片試料を作製する薄切片試料作製装置に有用である。 According to the thin-section sample evaluation apparatus and the thin-section sample preparation apparatus including the evaluation apparatus according to the present invention, the thin-section sample is irradiated with blue light, and diffusely reflected light from the surface of the thin-section sample is received and received. Image data is obtained for two or more different colors of light. Based on the obtained image data of two or more colors of light, the surface state of the thin slice sample can be detected and evaluated. Therefore, the present invention is useful for a thin-section sample preparation apparatus for preparing a thin-section sample used for physicochemical sample analysis or microscopic observation of biological samples.
１　試料ブロック
３ａ、３ｂ　照射部
４　撮像部
６　表面状態検出部
８　制御部
１０　薄切片試料評価装置
１１　試料
１２　包埋剤
２０　試料ブロック搬送部
３０　カッター部
３１　カッター
３２　薄切片試料
３３　供給リール
３４　キャリアテープ
３５、３６、３７、３８　ガイドローラ
３９　巻取リール
４０　試料ブロック保管部
５０　位置調整部
６０　載置部
６１、６２　ガイドローラ
６３　スライドガラス
６４　接着液
７０　伸展部
８０　スライドガラス搬送部
９０　制御部
１００　薄切片試料作製装置 DESCRIPTION OF SYMBOLS 1 Sample block 3a, 3b Irradiation part 4 Imaging part 6 Surface state detection part 8 Control part 10 Thin section sample evaluation apparatus 11 Sample 12 Embedding agent 20 Sample block conveyance part 30 Cutter part 31 Cutter 32 Thin section sample 33 Supply reel 34 Carrier Tape 35, 36, 37, 38 Guide roller 39 Take-up reel 40 Sample block storage unit 50 Position adjustment unit 60 Placement unit 61, 62 Guide roller 63 Slide glass 64 Adhesive liquid 70 Extension unit 80 Slide glass conveyance unit 90 Control unit 100 Thin section sample preparation equipment

Claims (9)

  1. 　試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りし、スライドガラスの上に載置した薄切片試料の表面状態を評価する薄切片試料評価装置であって、
    　前記スライドガラスの上に載置した前記薄切片試料の表面に青色光を照射する照射部と、
    　前記照射部から照射した光の前記薄切片試料の表面による拡散反射光を受光して異なる２色以上の光についての画像データを得る撮像部と、
    　前記撮像部で得られた前記２色以上の光についての前記画像データに基づいて、前記薄切片試料の表面状態を検出する表面状態検出部と、
    　少なくとも、前記撮像部と、前記表面状態検出部と、を制御し、前記表面状態検出部で検出した前記薄切片試料の表面状態について評価する制御部と、
    を備えた、薄切片試料評価装置。     A thin slice sample evaluation device for thinly slicing a sample block in which a sample is embedded in an embedding agent with a cutter and evaluating the surface state of the thin slice sample placed on a slide glass,
    An irradiation unit for irradiating the surface of the thin-section sample placed on the slide glass with blue light;
    An imaging unit that receives diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit and obtains image data of two or more different colors;
    A surface state detection unit that detects a surface state of the thin-section sample based on the image data about the light of two or more colors obtained by the imaging unit;
    A control unit that controls at least the imaging unit and the surface state detection unit, and evaluates the surface state of the thin-section sample detected by the surface state detection unit;
    A thin-section sample evaluation apparatus.
  2. 　前記照射部は、前記薄切片試料と、前記スライドガラスとの間に水分が存在する状態で、前記薄切片試料の表面に青色光を照射する、請求項１に記載の薄切片試料評価装置。 The thin-section sample evaluation apparatus according to claim 1, wherein the irradiation unit irradiates the surface of the thin-section sample with blue light in a state where moisture exists between the thin-section sample and the slide glass.
  3. 　前記撮像部は、前記薄切片試料の鉛直上方に配置され、
    　前記照射部は、前記撮像部を挟んで左右に配置される、請求項１に記載の薄切片試料評価装置。     The imaging unit is disposed vertically above the thin slice sample,
    The thin-section sample evaluation apparatus according to claim 1, wherein the irradiation unit is arranged on the left and right sides of the imaging unit.
  4. 　前記照射部は、直線照明である、請求項３に記載の薄切片試料評価装置。 The thin-section sample evaluation apparatus according to claim 3, wherein the irradiation unit is linear illumination.
  5. 　前記照射部は、前記撮像部を囲むように配置された環状照射部である、請求項３に記載の薄切片試料評価装置。 The thin-section sample evaluation device according to claim 3, wherein the irradiation unit is an annular irradiation unit arranged so as to surround the imaging unit.
  6. 　試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りして薄切片試料を作製する薄切片試料作製装置であって、
    　前記試料ブロックを搬送する試料ブロック搬送部と、
    　前記試料ブロックの表面をカッターによって薄切りして薄切片試料を得るカッター部と、
    　前記薄切片試料を、スライドガラスの上に載置する載置部と、
    　請求項１から５のいずれか一項に記載の前記薄切片試料評価装置と、
    　少なくとも、前記試料ブロック搬送部と、前記載置部と、前記薄切片試料評価装置と、を制御する制御部と、
    を備えた、薄切片試料作製装置。     A thin-section sample preparation device for preparing a thin-section sample by slicing a sample block in which a sample is embedded in an embedding agent with a cutter,
    A sample block transport unit for transporting the sample block;
    A cutter unit that slices the surface of the sample block with a cutter to obtain a thin slice sample;
    A placement section for placing the thin slice sample on a slide glass;
    The thin-section sample evaluation device according to any one of claims 1 to 5,
    At least a control unit that controls the sample block transport unit, the placement unit, and the thin-section sample evaluation device,
    A thin-section sample preparation apparatus comprising:
  7. 　前記薄切片試料評価装置によって、前記薄切片の表面状態に不具合を検出した場合に、前記カッター部の前記カッターを交換する、請求項６に記載の薄切片試料作製装置。 The thin-section sample preparation device according to claim 6, wherein when the defect is detected in the surface state of the thin section by the thin-section sample evaluation device, the cutter of the cutter unit is replaced.
  8. 　前記薄切片試料評価装置によって、前記薄切片の表面状態に不具合を検出した場合に、薄切片試料の作製を停止する、請求項６に記載の薄切片試料作製装置。 The thin-section sample preparation device according to claim 6, wherein when the thin-section sample evaluation apparatus detects a defect in the surface state of the thin section, the preparation of the thin-section sample is stopped.
  9. 　試料が包埋剤中に包埋された試料ブロックをカッターにより薄切りし、スライドガラスの上に載置した薄切片試料の表面状態を評価する薄切片試料評価方法であって、
    　前記スライドガラスの上に載置した前記薄切片試料の表面に青色光を照射する照射ステップと、
    　前記照射部から照射した光の前記薄切片試料の表面による拡散反射光を受光して、受光した光のうち異なる２色以上の光についての画像データを得る撮像ステップと、
    　前記撮像部で得られた前記２色以上の光についての前記画像データに基づいて、前記薄切片試料の表面状態を検出する表面状態検出ステップと、
    　前記検出した前記薄切片試料の表面状態について評価する評価ステップと、
    を含む、薄切片試料評価方法。     A thin slice sample evaluation method for evaluating a surface state of a thin slice sample placed on a slide glass by slicing a sample block in which a sample is embedded in an embedding agent,
    An irradiation step of irradiating the surface of the thin slice sample placed on the slide glass with blue light;
    An imaging step of receiving diffusely reflected light from the surface of the thin-section sample of the light irradiated from the irradiation unit and obtaining image data of light of two or more different colors among the received light;
    A surface state detection step of detecting a surface state of the thin-section sample based on the image data for the light of two or more colors obtained by the imaging unit;
    An evaluation step for evaluating the surface condition of the detected thin slice sample;
    A method for evaluating thin slice samples.
PCT/JP2014/075914 2013-09-30 2014-09-29 Sliced specimen evaluation device, sliced specimen preparation device including said sliced specimen evaluation device, and sliced specimen evaluation method WO2015046518A1 (en)

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