Observation Holder, Observation Device, Observation Wafer, and Method for Manufacturing the Same

Abstract

The present invention relates to an observation holder. The aforementioned observation holder includes: a housing portion that has a rectangular shape in a plan view and is configured to accommodate an observation object; and an observation portion provided at the lower part of the housing portion. It is characterized in that: the aforementioned observation portion is configured to be able to observe the observation object accommodated in the aforementioned housing portion from below. The aforementioned housing portion includes: a housing portion main body provided with a plurality of holes for accommodating the observation object separately from each other; and an upper part of the housing portion provided at the upper part of the aforementioned housing portion main body. The upper part of the housing portion includes: a wall portion surrounding the space above the aforementioned housing portion main body, a storage portion for storing a specified liquid. An opening is provided in the upper part of the aforementioned space of the upper part of the housing portion. The aforementioned plurality of holes are arranged in the horizontal direction, and each hole penetrates the aforementioned housing portion main body in the vertical direction and communicates with the aforementioned opening. Two adjacent holes among the aforementioned plurality of holes are connected in the aforementioned observation portion.

Description

Technical Field

[0001] The present invention relates to a holder for observation (mainly a holder for observing cells). Background Art

[0002] In Patent Documents 1 and 2, a structure of a holder for observation that observably houses an object to be observed is described. The holder for observation is configured such that a cell as an object to be observed can be observed from below the holder for observation. Users such as physicians, medical technicians, and researchers can use such a holder for observation and use a prescribed observation instrument (e.g., a microscope, a video camera, etc.) to observe the object to be observed by means of an image. Examples of the object to be observed include tissues or body fluids themselves that have cells as constituent elements in animals including humans, such as blood or lymph fluid, ascites washing fluid, alveolar washing fluid, various white blood cells or other cells as their constituent components, cancer cells isolated from tissues or cultured cells that have been established, in humans or mice; furthermore, examples include isolated plant cells including microorganisms such as fungi. A holder for observation that can simultaneously observe multiple different functions possessed by cells as such objects to be observed by means of an image is used for, for example, large-scale screening tests for neonates or the diagnosis of various diseases (Patent Document 3).

[0003] Prior Art Documents

[0004] Patent Documents

[0005] Patent Document 1: Japanese Patent Application Laid-Open No. 2007-124904

[0006] Patent Document 2: Japanese Patent Application Laid-Open No. 2011-038923

[0007] Patent Document 3: Japanese Patent Application Laid-Open No. 2018-102295 Summary of the Invention

[0008] Problems to be Solved by the Invention

[0009] In the above-described holder for observation, in order to be able to simply and efficiently observe multiple different characteristics and functions possessed by cells as numerous objects to be observed at one time, and further, to be able to simultaneously perform these observations and measurements on numerous samples, for example, multiple patient samples, a further improvement in the structure is sought.

[0010] An exemplary object of the present invention is to provide a novel structure of a holder for observation that can simply and efficiently observe multiple different characteristics possessed by one object to be observed, and further, can perform these observations on numerous samples at one time.

[0011] Means for Solving the Problems

[0012] One aspect of the present invention relates to a holder for observation. The aforementioned holder for observation includes: a housing portion that has a rectangular shape in a top view and is configured to accommodate an object to be observed; and an observation portion provided at the lower part of the housing portion. It is characterized in that the aforementioned observation portion is configured to be able to observe the object to be observed accommodated in the aforementioned housing portion from below. The aforementioned housing portion includes: a housing portion body provided with a plurality of holes for accommodating the object to be observed separately from each other; and an upper portion of the housing portion provided at the upper part of the aforementioned housing portion body. The upper portion of the housing portion includes a wall portion surrounding the space above the aforementioned housing portion body, serving as a reservoir for storing a prescribed liquid. An opening is provided in the upper portion of the aforementioned space in the upper portion of the housing portion. The aforementioned plurality of holes are arranged in the horizontal direction, and each hole penetrates the aforementioned housing portion body in the vertical direction and communicates with the aforementioned opening. Two adjacent holes among the aforementioned plurality of holes are connected at the aforementioned observation portion.

[0013] Advantages of the Invention

[0014] According to the present invention, a plurality of different characteristics of one object to be observed can be observed simply and efficiently.

[0015] Other features and advantages of the present invention can be clearly understood from the following description with reference to the drawings. In addition, in the drawings, the same or similar components are given the same reference numerals. Description of the Drawings

[0016] The drawings are included in the specification and form a part of the specification, showing embodiments of the present invention, and are used together with the description to explain the principle of the present invention.

[0017] Figure 1 It is a schematic side view showing a configuration example of an observation device.

[0018] Figure 2 It is a perspective view of a part of the observation device.

[0019] Figure 3 It is a perspective view of the holder for observation.

[0020] Figure 4 It is an exploded view of the holder for observation.

[0021] Figure 5 It is a perspective view of a plate material that is a part of the observation portion.

[0022] Figure 6 It is a perspective view of the lower side of the holder for observation.

[0023] Figure 7A It is a sectional view of the holder for observation.

[0024] Figure 7B It is a sectional view of the holder for observation.

[0025] Figure 8A It is a diagram for explaining the manufacturing method of the observation unit.

[0026] Figure 8B It is a diagram for explaining the manufacturing method of the observation unit.

[0027] Figure 8C It is a diagram for explaining the manufacturing method of the observation unit.

[0028] Figure 8D It is a diagram for explaining the manufacturing method of the observation unit.

[0029] Figure 8E It is a diagram for explaining the manufacturing method of the observation unit.

[0030] Figure 8F It is a diagram for explaining the manufacturing method of the observation unit.

[0031] Figure 8G It is a diagram for explaining the manufacturing method of the observation unit.

[0032] Figure 8H It is a diagram for explaining the manufacturing method of the observation unit.

[0033] Figure 8I It is a diagram for explaining the manufacturing method of the observation unit. Detailed implementation manners

[0034] Hereinafter, the implementation manners will be described in detail with reference to the drawings. In addition, the following implementation manners do not limit the invention according to the claims. Further, all combinations of the features described in the implementation manners are not necessarily essential for the invention. Two or more of the features described in the implementation manners can be arbitrarily combined. In addition, the same or similar components are given the same reference numerals, and repeated descriptions are omitted.

[0035] (Regarding the overall configuration of the observation device)

[0036] Figure 1 It shows a schematic side view of the observation device 1 according to the implementation manner, Figure 2 It shows a perspective view of a part of the observation device 1. The observation device 1 includes a support table 11, a imaging unit 12, and a moving mechanism 13. In addition, the observation device 1 includes an observation preparation unit 14. Further, the observation holder 2, which will be described later and is not included in the observation device 1, is placed on the support table 11.

[0037] The support table 11 is a plate-like member with a specified thickness. The support table 11 is configured to support a plurality of observation holders 2, and the plurality of observation holders 2 are arranged in a horizontal direction on the support table 11. The horizontal direction referred to herein is any direction parallel to the upper surface of the support table 11 when the support table 11 is formed in a horizontal posture. Each observation holder 2 is configured to accommodate a specified observation object, and the details will be described in detail later. As an example of the observation object, for example, white blood cells in animals such as human blood are listed.

[0038] In addition, in Figure 2 one direction in the arrangement direction of the plurality of observation holders 2 is represented as the X direction, and in addition, the other direction (the direction orthogonal to the X direction) in the arrangement direction is represented as the Y direction. In addition, the above arrangement direction is taken as the horizontal direction, and the vertical direction or the up-and-down direction is represented as the Z direction.

[0039] In addition, in the present embodiment, six concave portions 111 are arranged at equal intervals in the X direction and two rows are arranged in the Y direction on the support table 11. In each concave portion 111, the observation holder 2 can be fitted and placed. In addition, at the bottom 111a of each concave portion 111, an opening 112 penetrating in the up-and-down direction is provided, and the placed observation holder 2 can be observed from below.

[0040] The bottom 111a of the concave portion 111 forms an opening 112, and a placement portion 111b is formed by extending toward the center side in a flange shape along its edge, whereby the observation holder 2 can be placed. The plate material (second plate material) 222 of the observation portion 22 of the observation holder 2 to be described later abuts against the placement portion 111b and is placed. In addition, the side wall of the concave portion 111 constitutes a peripheral wall (guide wall) 111c, and the peripheral wall 111c is used to restrict the horizontal movement of the observation holder 2 when the observation holder 2 is fitted. For the height of the peripheral wall 111c, for example, it is formed in a size higher than the lower part 210L of the main body of the accommodation portion 210 of the observation holder 2 to be described later and lower than the upper part of the main body of the accommodation portion 210.

[0041] The imaging unit 12 is configured to image the observation object accommodated in the observation holder 2, and is provided below the support table 11 in the present embodiment. For the imaging unit 12, any device that can image a dynamic image and / or a still image, such as a known camera equipped with a CCD / CMOS image sensor, can be used. In addition, in the following description, the imaging performed by the imaging unit 12 can also be expressed as observation.

[0042] The moving mechanism 13 is configured to relatively move the imaging unit 12 horizontally with respect to the support table 11, and the moving direction is substantially parallel to the X direction in the present embodiment. When the imaging unit 12 is moved by the moving mechanism 13, the imaging unit 12 moves facing each of the openings 112 arranged in a plurality in the X direction. For the moving mechanism 13, a known sliding mechanism can be used. For example: a guide rail 131 that slidably supports the imaging unit 12 in the X direction, a nut (not shown) included in the imaging unit 12, a ball screw that is arranged in parallel along the guide rail 131 and is rotatably supported by being screwed with the nut, an electric motor 132 that rotationally drives the ball screw, etc.

[0043] In the present embodiment, the plurality of observation holders 2 are as Figure 2 shown, and include a total of 12 recesses 111 arranged in two columns in the Y direction with 6 recesses 111 arranged in a row in the X direction as one column. The moving mechanism 13 can move the imaging unit 12 in the X direction and the Y direction. Alternatively, the moving mechanism 13 can also move the imaging unit 12 only in the X direction, and two sets of combinations of the moving mechanism 13 and the imaging unit 12 are provided in the Y direction. In addition, as the imaging unit 12, a vision camera, an objective lens, and other imaging devices are listed. When an objective lens is used as the imaging unit 12, the imaging unit 12 can also be formed as an index table, and a plurality of objective lenses with different magnifications are provided on the index table, and an objective lens with an arbitrary magnification can be selected by indexing rotation.

[0044] The observation preparation unit 14 is a unit for the user to perform preparation operations. The observation preparation unit 14 includes a preparation support table 141 and a moving mechanism (second moving mechanism) 142. The preparation support table 141 is configured to support the observation holder 2 in a state before accommodating the observation object (unaccommodated state). In the preparation support table 141, recesses 111 and openings 112 are provided in the same manner as in the support table 11. Although it will be described in detail later, the user uses a prescribed instrument or device in the observation holder 2 supported on the preparation support table 141, and injects and accommodates the observation object by means of, for example, a manual injector formed by a syringe, a dropper, etc., or an automatic injector formed by a vertical 6-axis robot, a SCARA robot, etc., thereby performing the preparation for observation.

[0045] In addition, as the user mentioned here, typical examples include operators such as physicians, inspection technicians, and researchers, and can also be industrial robots or manipulators that operate according to them or their instructions.

[0046] The moving mechanism 142 is configured to move the standby support table 141 in a specified direction (at least the Z direction). For example, when using the standby support table 141, the moving mechanism 142 can be actuated to adjust the height of the standby support table 141 and make preparations for observation. For the moving mechanism 142, a known sliding mechanism can be used. For example, a guide rail 1421 that slidably supports the standby support table 141, an electric motor 1422 that slides the standby support table 141 along the guide rail 1421, etc.

[0047] In such an observation device 1, the user injects the object to be observed into the observation holder 2 supported by the standby support table 141 to prepare the object to be observed. Details will be described later. After that, by transferring the observation holder 2 from the standby support table 141 to the support table 11, the setting of the observation holder 2 relative to the observation device 1 is completed. Then, the imaging unit 12 observes the object to be observed from below the observation holder 2. Here, the preparation of the object to be observed means forming a state where the imaging unit 12 can image the observation unit 22 (described later) of the observation holder 2 from below, confirm the captured image, and at the same time inject the object to be observed into a specified position (observation position) of the observation holder 2 so that the cells that are the objects to be observed are arranged at the observation position. The observation device 1 can also be referred to as an observation system, an evaluation device, an evaluation system, a diagnostic device, a diagnostic system, etc.

[0048] (Regarding the structure of the observation holder)

[0049] Figure 3 A perspective view showing the observation holder 2. Figure 4 An exploded view showing the observation holder 2. The observation holder 2 includes a housing portion 21 and an observation portion 22. In addition, in Figures 3 - 4 corresponds to the X direction, Y direction, and Z direction. Figure 2 is shown.

[0050] The housing portion 21 has a rectangular shape when viewed from above (the viewpoint in the Z direction. Top view.). Here, it has a rectangular shape and is configured to house the aforementioned object to be observed. The housing portion 21 includes a housing body 210 and a housing upper portion 211 in the present embodiment. The housing body 210 has a protrusion 2101 at the center of the short side direction ( Figure 3 is the Y direction in) when viewed from above. The protrusion 2101 extends in the long side direction ( Figure 3 is the X direction in) when viewed from above. As a result, stepped surfaces 2102 are formed on both sides of the protrusion 2101.

[0051] At the protruding portion 2101 of the housing portion main body 210, a plurality of holes H1 for housing an object to be observed are provided separately from each other. The plurality of holes H1 are arranged in the X direction and the Y direction, and each hole H1 is provided so as to penetrate the protruding portion 2101 in the Z direction. For each surface of the plurality of holes H1 formed in the housing portion main body 210 and in contact with the liquid, a hydrophilic processing treatment is performed. The upper surface 210U of the protruding portion 2101 is flattened, and the opening end of each hole H1 is located on the upper surface 210U. In addition, the region including each hole H1 and its periphery in the upper surface 210U is made into a flat region 210R (see Figure 3 ).

[0052] The upper housing portion 211 is provided on the upper portion of the housing portion main body 210, and has an opening 2112 for forming a space above the flat region 210R of the housing portion main body 210 when the upper housing portion 211 is provided on the housing portion main body 210. In other words, the space surrounded by the wall portion 2111a in the upper housing portion 211 is the opening 2112. The opening 2112 is formed to penetrate the upper housing portion 211, and the opening area is different between the upper portion and the lower portion of the upper housing portion 211. The lower opening 2112b is the portion into which the protruding portion 2101 is inserted, and is formed in a size substantially the same as the shape of the protruding portion 2101.

[0053] On the other hand, the upper opening 2112a is the portion for forming a space above the flat region 210R, and is formed in a size substantially the same as the shape of the flat region 210R. That is, the opening area of the upper opening 2112a is smaller than the opening area of the lower opening 2112b. A part of the upper surface 210U of the protruding portion 2101 is engaged with the stepped surface SS between the upper opening 2112a and the lower opening 2112b, and as a result, the flat region 210R faces the upper opening 2112a. At this time, a part of the lower portion 211L of the upper housing portion 211 is engaged with the stepped surface 2102 of the housing portion main body 210. In this state, the space surrounded by the upper opening 2112a and the flat region 210R functions as a storage portion for storing a predetermined liquid. In addition, the uppermost portion 2112c of the opening 2112 is formed to expand in a conical shape.

[0054] In this way, the cross-sectional area of the opening 2112 is different between the upper portion and the lower portion of the upper housing portion 211, and the storage portion is formed by the upper opening 2112a having a small cross-sectional area to reduce the storage volume. Thus, the amount of liquid stored in the storage portion can be reduced. In addition, for each surface of the upper housing portion 211 formed as the storage portion and in contact with the stored liquid, a hydrophilic processing treatment is performed in the same manner as the surface where the plurality of holes H1 are formed. As the liquid, a liquid medium corresponding to the object to be observed is used, but other solutions such as physiological saline may also be used.

[0055] The multiple holes H1 provided in the housing part main body 210 are not used independently of each other. Instead, two adjacent holes H1 are taken as a group and used as a U-shaped passage formed by connecting the lower ends of the holes H1 to each other in the observation part 22.

[0056] In the present embodiment, the housing part main body 210 is made of a non-light-transmitting material. In addition, the upper part 211 of the housing part is made of a light-transmitting material. In addition, the so-called non-light-transmitting means that the transmittance of visible light is relatively low. For example, the transmittance may be 50% or less, preferably 30% or less, and more preferably 10% or less. In addition, the so-called light-transmitting means that the transmittance of visible light is relatively high. For example, the transmittance may be 50% or more, preferably 70% or more, and more preferably 90% or more. As another embodiment, the upper part 211 of the housing part may also be made of a non-transmitting material.

[0057] In addition, the housing part main body 210 and the upper part 211 of the housing part may also be formed of a single member. In the case of being formed of a single member, it is preferably made of a non-transmitting material.

[0058] The observation part 22 is provided in the lower part of the housing part 21 and is configured to be able to observe an object to be observed housed in the housing part 21 from below by the imaging part 12. The observation part 22 includes a plate material (first plate material) 221 and a plate material (second plate material) 222. The plate material 221 covers the housing part main body 210 from below. In addition, the plate material 222 covers the plate material 221 from below. The plate material 222 covers the plate material 221 from below and is substantially made of a light-transmitting material. The plate material 221 is partially made of a light-transmitting material in the present embodiment, but may also be entirely made of a non-light-transmitting or reflective material. In addition, the surface may also use a material of a dark color system (such as black). The plate material 221 uses, for example, a silicon wafer (silicon wafer) microfabricated by anisotropic dry etching (inductively coupled plasma reactive ion etching). The plate materials 221 and 222 are rectangular members of the same size when viewed from above in the present embodiment. As long as at least one of the plate materials 221 and 222 is configured such that the two adjacent holes H1 communicate with each other. Next, while referring to Figure 5 , an explanation will be given here.

[0059] Figure 5A perspective view showing the plate material 221. In the present embodiment, in the plate material 221, a plurality of through holes H3 corresponding to the plurality of holes H1 are provided. On the lower surface of the plate material 221, a recess CC1 that connects two (for example, H3a and H3b) of the plurality of through holes H3 adjacent to each other is provided. The recess CC1 is provided such that its outer edge surrounds the two through holes H3. That is, the plurality of through holes H3 are independently provided at positions facing the plurality of holes H1 on the upper surface side of the plate material 221. In contrast, on the lower surface side of the plate material 221, the two adjacent through holes H3 communicate with each other by means of the recess CC1, and thus become partially non-independent. In the case of the present embodiment, the recess CC1 is provided in an oval shape when viewed from above. The recess CC1 may also be referred to as a terrace, a recess, etc.

[0060] The recess CC1 includes a flat portion PL between the two adjacent through holes H3. The flat portion PL includes a flat surface PL1. In addition, the outer edge of the recess CC1 is constituted by a side wall WP. The side wall WP includes peripheral wall portions WP1, WP2, and two WP3s. The peripheral wall portions WP1 and WP2 are wall portions corresponding to the circumferential portions in the oval, and the peripheral wall portion WP3 is a wall portion corresponding to the straight line portion in the oval. In addition, the peripheral wall portion WP1 is a portion facing one of the two through holes H3 (here, H3a), and the peripheral wall portion WP2 is a portion facing the other of the two through holes H3 (here, H3b).

[0061] Specifically, the peripheral wall portion WP1 includes: a cylindrical upper peripheral wall portion WP1U formed with the same size as the size of the through hole H3; and a semi-cylindrical lower peripheral wall portion WP1L formed continuously with the upper peripheral wall portion WP1U and formed with the same size. In addition, the peripheral wall portion WP2, like the peripheral wall portion WP1, includes: a cylindrical upper peripheral wall portion WP2U formed with the same size as the size of the through hole H3; and a semi-cylindrical lower peripheral wall portion WP2L formed continuously with the upper peripheral wall portion WP2U and formed with the same size.

[0062] The two peripheral wall portions WP3 are respectively provided to form a continuous surface connected to the peripheral wall portions WP1 and WP2. Specifically, the two peripheral wall portions WP3 are respectively provided as a continuous surface that connects the lower peripheral wall portion WP1L of the peripheral wall portion WP1 and the lower peripheral wall portion WP2L of the peripheral wall portion WP2 and has the same height as the lower peripheral wall portion WP1L and the lower peripheral wall portion WP2L, wherein the lower peripheral wall portion WP2L of the peripheral wall portion WP2 is provided facing the lower peripheral wall portion WP1L of the peripheral wall portion WP1.

[0063] In the present embodiment, in a plan view, each through hole H3 is formed to be substantially circular, and the upper peripheral wall portions WP1U and WP2U of the peripheral wall portions WP1 and WP2 are each formed to depict a circle having the same diameter as the diameter of the through hole H3. In addition, the lower peripheral wall portions WP1L and WP2L of the peripheral wall portions WP1 and WP2 are each formed to depict an arc of a semi-circle having the same diameter as the diameter of the through hole H3, and the peripheral wall portion WP3 is formed to depict a substantially straight line.

[0064] A flat plate-like material 222 is used to cover the lower surface of the material 221 formed as described above, that is, the recess CC1, thereby forming a passage formed by the space surrounded by the peripheral wall portions WP1, WP2, WP3, the flat portion PL, and the material 221 as a connection passage 220W (see Figure 6 ). Thus, the lower portions of two adjacent through holes H3 are connected to each other via the connection passage 220W, and the two through holes H3 are connected to form one passage. Also, on the upper surface side of the material 221, the respective through holes H3 corresponding to two adjacent holes H1 are made to communicate with each other, so that when an object to be observed accommodated in the accommodation portion 21 stays in the connection passage 220W, observation can be performed using the imaging unit 12. In addition, a hydrophilic processing is performed on each surface of the recess CC1 that forms the connection passage 220W connecting a plurality of adjacent holes H1 and that is in contact with the liquid moving in the connection passage 220W.

[0065] In addition, the shape of the side wall WP is not limited to the present embodiment, as long as it is formed to include all of the above two through holes H3 in a plan view. In addition, although the size (diameter) of the through hole H3, the size (diameter) of WP1, and the size (diameter) of WP1 are preferably such that the respective arcs are aligned (are of the same size), as long as there is no influence such as making it difficult to see during observation, they may have different sizes from each other.

[0066] Referring again to Figure 4 , in the main body lower portion 210L of the accommodation portion main body 210, a regulating portion 210a for regulating the position of the material 221 is provided. In the present embodiment, the regulating portion 210a is a recessed portion recessed in the bottom surface of the accommodation portion main body 210, and in a plan view, the shape of the recess follows the outer shape of the material 221. The depth of the recessed portion is set to be equal to or greater than the thickness of the material 221. By fitting the material 221 of the observation portion 22 into the recess serving as the regulating portion 210a, the positioning of the observation portion 22 with respect to the accommodation portion 21 can be appropriately performed. In practice, as long as the upper peripheral wall portions WP1U and WP2U of the peripheral wall portions WP1 and WP2 corresponding to the through hole H3 can be appropriately aligned. Alternatively or additionally, a prescribed mark such as printing or a notch may be provided as the regulating portion 210a.

[0067] Figure 6 A bottom perspective view of the observation holder 2 formed by assembling the aforementioned housing portion 21 and the observation portion 22. The plate material 222 of the observation portion 22 is made of a light-transmissive material. Therefore, when the object to be observed stays in the connection path 220W connecting the two through-holes H3 (for example, H3a and H3b), imaging can be performed via the observation portion 22 by the imaging portion 12 (see Figures 1 - 2 ).

[0068] Regarding the flat surface PL1 that forms a part of the connection path 220W, it is preferable to set the entire area between the two through-holes H3 (for example, H3a and H3b) as the flat surface. However, the flat surface PL1 included in the flat portion PL may also be provided only in a part of this area. In this case, there may also be steps partially in this area (at positions where the flat surface PL1 has a different height from other flat portions PL).

[0069] Figure 7A A sectional view of the observation holder 2 with a vertical plane passing through the plurality of holes H1 and along the X direction as the section. Figure 7B Indicates Figure 7A A sectional view taken along line VIIB-VIIB of Figure 7B In addition, a partial enlarged view including the observation portion 22 surrounded by a dashed line. The observation portion 22 is configured such that two adjacent holes H1 among the plurality of holes H1 provided in the housing body 210 are connected via the corresponding through-holes H3a and H3b and the connection path 220W, and a group of holes H1 forms one path.

[0070] As an example, in the case where the object to be observed is blood, the holes H1 and the through-holes H3 may be set to have a diameter of, for example, about 700 [μm] to 1300 [μm], preferably about 900 [μm] to 1100 [μm], and more preferably about 1000 [μm]. In addition, the distance between the two through-holes H3 may be set to, for example, about 350 [μm] to 650 [μm], preferably about 450 [μm] to 550 [μm], and more preferably about 500 [μm]. In this case, the recess CC1 may be determined according to the particle size present in the body fluid or the like used. For example, in the case of human blood, according to the size of the white blood cells present therein, it may be set to a depth of, for example, about 2 [μm] to 7 [μm], preferably about 3 [μm] to 5 [μm], and more preferably about 4 [μm]. In addition, in the case of cancer cells separated from tissues, etc., it may be set to a depth of, for example, about 12 [μm] to 5 [μm], preferably about 10 [μm] to 6 [μm], and more preferably about 8 [μm]. In addition, when the flat surface PL1 including the stepped portion is provided on the flat portion PL, the depth of the flat portion PL is set to 50 - 100 [μm], and the distance between the flat surface PL1 and the surface of the plate 222 forming the connection path 220W is less than the particle size present in the body fluid or the like used, and it may be set at a distance allowing the particles to move.

[0071] Here, in order to appropriately achieve the observation of the object to be observed by the imaging unit 12, the imaging unit 12 may be set to scan directly below the flat surface PL1 in the recess CC1 between the two through-holes H3 (for example, H3a and H3b) with the +Z direction as the center of the pointing range.

[0072] In addition, by Figures 7A - 7B It is also known that in the present embodiment, the housing body 210 and the housing upper portion 211 are configured such that their widths in one horizontal direction (here, the Y direction) are equal to each other. Thereby, miniaturization of the housing portion 21, that is, the observation holder 2 itself can be achieved, and a plurality of observation holders 2 can be densely arranged on the support table 11. In the present embodiment, particularly dense arrangement can be performed in one horizontal direction (here, the Y direction).

[0073] In addition, in the present embodiment, the housing portion main body 210 and the upper housing portion 211 are configured such that their widths in the other horizontal direction (here, the X direction) are different from each other. With such a configuration, it is possible to reduce the size of the observation portion 22 of the recess CC1 that is set as the imaging range of the imaging unit 12, the housing portion main body 210 provided with a plurality of holes H1, and the upper housing portion 211 provided with the opening 2112, and it is possible to reduce the manufacturing material of the observation holder 2. In addition, it is possible to prevent the user from failing to hold the observation holder 2 (such as the observation holder 2 falling as a result). In addition, although these widths may be different from each other in one of the X direction and the Y direction, as another embodiment, they may be equal to each other in both directions.

[0074] In the observation holder 2 of the present embodiment, as Figure 3 shown, an information supply portion 23 is provided on the upper housing portion 211. Specifically, the information supply portion 23 is provided on the upper surface 211U of the upper housing portion. The information supply portion 23 can supply information about the observation object. This information may also be expressed as observation object information. As an example, the name of the subject, the collection date of the observation object, etc. are listed. In addition, this information may also be code information managed in association with the name of the subject, the collection date of the observation object, etc. As the code information, number information obtained by combining a plurality of numbers, two-dimensional data information (for example, barcodes, QR codes), etc. can be used. The code information can be read by a code information reading device (not shown). The information supply portion 23 may also be configured to be able to print the observation object information, or may be configured to be able to paste a sticker on which the observation object information is written. By using the information supply portion 23, the user can manage the observation object relatively easily.

[0075] (Regarding the method of housing and observing the observation object)

[0076] The user can access any of the holes H1 through the opening H2 using a prescribed instrument such as a syringe or a dropper in the observation holder 2 supported by the preparation support table 141, and can house the observation object in each of the holes H1. More specifically, first, a prescribed liquid (liquid medium) is filled in the space SP1 and the holes H1, and then the observation object (such as blood) is rapidly injected into one of two adjacent holes (a set of holes) H1 connected to the observation portion 22. As a result, a part of it flows into the connection passage 220W and the recess CC1. When a sufficient amount of cells required for observation do not flow into the recess CC1, negative pressure is applied to the other hole H1 side, whereby the cells can be made to flow into the recess CC1. Here, from Figure 3 it is also known that the uppermost portion 2112c of the opening 2112 can be set to form an inclined surface, for example. Thus, the user can appropriately access any of the holes H1 from the opening 2112.

[0077] The observation holder 2 that has finished accommodating the object to be observed is transferred to the support table 11, and the object to be observed can be observed from below the observation holder 2 via the observation unit 22 using the imaging unit 12.

[0078] As described above, in the present embodiment, the main body 210 of the accommodating portion is made of a non-translucent material, and the upper portion 211 of the accommodating portion is made of a translucent material. Since the main body 210 of the accommodating portion is made of a non-translucent material, the contrast between the flat surface PL1 that is the observation target portion and the through hole H3 that is the non-observation target portion can be made more distinct, and it is easy to observe the object to be observed injected into the flat surface PL1 portion that is the observation target portion from below via the observation unit 22 using the imaging unit 12. In addition, since the upper portion 211 of the accommodating portion is made of a translucent material, when the object to be observed is accommodated in the hole H1, the user can observe the inside of the opening 2112 and each hole H1 from both above and side.

[0079] As another embodiment, instead of the user using a specified instrument such as a syringe or a dropper to access any hole H1 through the opening 2112 and perform the operation of accommodating the object to be observed in each hole H1, it is also possible to automatically accommodate the object to be observed by means of an automatic machine. In this case, the upper portion 211 of the accommodating portion may not be made of a transmissive material, and both the main body 210 of the accommodating portion and the upper portion 211 of the accommodating portion may be made of a non-translucent material. By making both the main body 210 of the accommodating portion and the upper portion 211 of the accommodating portion made of a non-translucent material, the position of each hole H1 can be observed from above using the detection mechanism (such as a camera) included in the automatic machine, and the operation of accommodating the object to be observed can be performed efficiently.

[0080] Refer again to Figure 2 , in the support table 11 that supports the observation holder 2, there are provided a recess 111 for fitting and placing the observation holder 2, and an opening 112 that penetrates the bottom of the recess 111 in the vertical direction. By supporting the lower surface of the peripheral edge of the plate material 222 by the placement portion 111b of the recess 111, the height up to the flat surface PL1 portion that becomes the observation target portion can be kept constant, and the observation from below the object to be observed using the imaging unit 12 can be appropriately achieved. The same applies to the preparation support table 141.

[0081] The observation holder 2 can be discarded after the observation of the object to be observed is completed, and can also be called a disposable holder or the like. That is, by forming the observation holder 2 to be disposable (it is recommended to be discarded after one use), it is possible to further prevent the mixing of objects to be observed different from the measurement sample, so-called contaminants. Therefore, the observation holder 2 can also be configured to unitize its entire body. For example, the housing part 21 and the observation part 22 (each element constituting them) can also be inseparably fixed, and in the housing part 21, the housing body 210 and the upper part 211 of the housing part can also be inseparably fixed.

[0082] In the present embodiment, the observation holder 2 as a disposable holder is respectively configured with a corresponding shape or outer shape to form a recess CC1 in the observation part 22, a plurality of holes H1 in the housing body 210, and an opening 2112 in the upper part 211 of the housing part, thereby achieving miniaturization and being effectively utilized flexibly.

[0083] (Regarding the manufacturing method of the observation part)

[0084] As described above, the above-mentioned observation part 22 including the plates 221 and 222 is provided at the lower part of the housing part 21. Thus, the housing part 21 and the observation part 22 are formed into a container. From this perspective, the observation part 22 can also be called the bottom of the housing part, the bottom of the container, etc., and the observation holder 2 can also be called an observation container or the like. For such an observation part 22, an observation wafer that can be installed as a component in the observation holder 2 can be prepared separately, and can be fabricated using known semiconductor manufacturing techniques.

[0085] Figures 8A - 8I It is a diagram for explaining each process of the manufacturing method of the observation part 22. The outline of this manufacturing method is to overlap the plate 222 on the lower surface of the plate 221 and then bond the plates 221 and 222 by anodic bonding. In addition, hereinafter, for the sake of easy explanation, the main processes in this manufacturing method will be emphasized in the description, but cleaning treatment, heat treatment, etc. can be appropriately performed as needed.

[0086] In Figure 8A the process, a semiconductor substrate SB1 that becomes the material of the plate 221 is prepared, and the semiconductor substrate SB1 is oxidized. For the semiconductor substrate SB1, a silicon substrate can be suitably used. In addition, the film thickness of the oxide film OX1 formed in this process is 0.12 μm to that extent.

[0087] In Figure 8B the process, in Figure 8AEtch a part of the upper surface of the structure obtained in the

[0088] In the Figure 8C process, oxidize the upper surface of the structure obtained in the Figure 8B process with a new oxide film OX1 covering the etched portion and the already existing oxide film OX1. Through this process, a semiconductor substrate SB1 with steps on its upper surface and an oxide film OX1 covering the upper surface can be obtained. Also, after the oxidation process, the upper surface of the oxide film OX1 can be planarized by performing a polishing process such as CMP (Chemical Mechanical Polishing).

[0089] In the Figure 8D process, form a metal film M1 on the upper surface of the structure obtained in the Figure 8C process. This process can be carried out by a known deposition method such as sputtering. Aluminum can be appropriately used as the material of the metal film M1. Additionally, the film thickness of the metal film M1 formed in this process is about 0.4 μm.

[0090] In the Figure 8E process, etch a part of the upper surface of the structure obtained in the Figure 8D process, and partially remove the metal film M1 and the oxide film OX1 to expose a part of the upper surface of the semiconductor substrate SB1, and then partially remove the exposed semiconductor substrate SB1.

[0091] In the Figure 8F process, perform a further etching process on the etched portion formed in the Figure 8E process, and deepen the depth of the etched portion formed in the Figure 8E process. As a result, the etched portion reaches near the oxide film OX1 on the lower surface side in the semiconductor substrate SB1.

[0092] In the Figure 8G process, perform a polishing process such as CMP on the lower surface side of the structure obtained in the Figure 8F process. As a result, the etched portion becomes a through hole H3 penetrating the semiconductor substrate SB1.

[0093] In the Figure 8H process, from the Figure 8GIn the structure obtained in the process, the metal film M1 and the oxide film OX1 are removed. Additionally, a surface oxidation treatment is incidentally performed to form an oxide film (not shown). Thus, the obtained structure is formed into the plate material 221.

[0094] In Figure 8I the process, the plate material 222 is overlapped on the plate material 221 obtained in Figure 8H the process, and they are joined by anodic bonding. For the material of the plate material 222, a material with a transparent and flatness-guaranteed property is used. Alternatively, depending on the material used, they can be joined by other appropriate joining methods such as solid-phase bonding. The observation unit 22 can be fabricated in the above sequence.

[0095] The observation wafer that becomes the above-mentioned observation unit 22 is actually obtained by collectively manufacturing multiple plate materials 221 at once. Since multiple observation wafers are manufactured at once, the above-mentioned processing for manufacturing multiple plate materials 221 is performed on a first mother plate (here, a silicon substrate) of a specified size that can handle multiple plate materials 221. Next, a second mother plate (here, a glass plate) of the same size as the first mother plate is overlapped and joined. After that, through a cutting process or the like that divides the joined first and second mother plates into individual wafers, multiple observation wafers can be manufactured at once.

[0096] In addition, when manufacturing different types of observation wafers having a flat surface PL1 including steps in the flat portion PL, there is a risk of generating shadows at the boundary between the flat surface PL1 and other surfaces of the flat portion PL. Therefore, it is preferable to perform right-angle machining so that the stepped wall surfaces formed as steps are perpendicular to each surface. This can be achieved by the same manufacturing method as the above-mentioned manufacturing method. However, in this case, in order to machine the boundary portion into a right angle, it is preferable to use a silicon substrate (silicon wafer) having a thickness capable of performing right-angle machining.

[0097] (Summary)

[0098] According to the present embodiment, the observation holder 2 includes a housing portion 21 and an observation portion 22. The housing portion 21 is configured to have a rectangular shape in plan view and can house an object to be observed (e.g., blood). The observation portion 22 is provided below the housing portion 21 and is configured to be able to observe the object to be observed housed in the housing portion 21 from below. The housing portion 21 includes a housing body 210 and a housing upper portion 211 provided on its upper part. In the housing body 210, a plurality of holes H1 for housing the object to be observed are provided. The housing upper portion 211 includes a wall portion 2111a surrounding the upper opening 2112 of the housing body 210. The wall portion 2111a functions as a storage portion or a blocking portion for storing a prescribed liquid (liquid medium) in the opening 2112. The plurality of holes H1 are arranged in the horizontal direction, and each hole H1 penetrates the housing body 210 in the vertical direction and communicates with the opening 2112. In addition, two adjacent ones of the plurality of holes H1 are connected at the observation portion 22.

[0099] According to such a configuration, the object to be observed located between two adjacent holes H1 can be observed from below via the observation portion 22. Thus, when a plurality of observation holders 2 are arranged on a prescribed support table 11, a large number of objects to be observed or a plurality of different characteristics of one object to be observed can be observed simply and efficiently at one time. In addition, by performing a hydrophilic processing on each surface in contact with the liquid, the storage of the liquid or the injection of the sample and the arrangement of the sample at the observation position can be easily and reliably performed. In addition, since any of the plurality of holes H1 can be accessed from the opening H2, users such as physicians, medical technicians, and researchers can relatively easily house the object to be observed in any of the holes H1.

[0100] In the above description, for ease of understanding, each element is represented by a name associated with its function, but each element is not limited to having the content described in the embodiment as the main function, and may also have the content as an auxiliary function.

Claims

1. An observation holder (2) comprising: a housing portion (21) having a rectangular shape in plan view and configured to house an object to be observed, and an observation portion (22) provided at a lower portion of the housing portion (21), characterized in that, the observation portion (22) is configured to observe the object to be observed housed in the housing portion (21) from below, the housing portion (21) includes: a housing portion body (210) provided with a plurality of holes (H1) for housing an object to be observed separately from each other; and a housing portion upper part (211) provided at an upper portion of the housing portion body (210), the housing portion upper part (211) includes a wall portion (2111a) surrounding an upper space of the housing portion body (210) and serving as a storage portion for storing a specified liquid, an opening (2112) is provided at an upper portion of the space in the housing portion upper part (211), the plurality of holes (H1) are arranged in a horizontal direction, each hole (H1) penetrates the housing portion body (210) in a vertical direction and communicates with the opening (2112) located above the housing portion body (210) via the space, two adjacent holes among the plurality of holes (H1) are connected at the observation portion (22), whereby between the two adjacent holes, the object to be observed can be observed from a lower side of the two adjacent holes via the observation portion (22), the observation portion (22) includes: a first plate member (221) covering the housing portion body (210) from below; and a second plate member (222) made of a light-transmissive material covering the first plate member (221) from below, a plurality of through holes (H3) corresponding to the plurality of holes (H1) are provided in the first plate member (221), a recess (CC1) connecting two adjacent through holes among the plurality of through holes (H3) is provided on a lower surface of the first plate member (221), and an outer edge of the recess (CC1) surrounds the two through holes.

2. The observation holder (2) according to claim 1, characterized in that, the wall portion (2111a) surrounds the plurality of holes (H1) in a rectangular shape along an arrangement direction of the plurality of holes (H1).

3. The observation holder (2) according to claim 1, characterized in that, the housing portion body (210) is made of a non-light-transmissive material, the housing portion upper part (211) is made of a light-transmissive material.

4. The observation holder (2) according to claim 1, characterized in that, both the housing portion body (210) and the housing portion upper part (211) are made of a non-light-transmissive material.

5. The observation holder (2) according to claim 1, characterized in that, at least one of the first plate member (221) and the second plate member (222) is configured such that the two adjacent holes communicate with each other.

6. The observation holder (2) according to claim 5, characterized in that, the recess (CC1) forms a flat surface between the two adjacent through holes (H3).

7. The observation holder (2) according to claim 5, wherein: the concave portion (CC1) is provided in an oval shape when viewed from above; the side wall (WP) forming the concave portion (CC1) includes: a first peripheral wall portion (WP1) that is provided along a part of the outer edge of one of the two adjacent through holes, corresponding to the circumferential portion of the oval; a second peripheral wall portion (WP2) that is provided along a part of the outer edge of the other of the two adjacent through holes, corresponding to the other circumferential portion of the oval; and a third peripheral wall portion (WP3) that connects the first peripheral wall portion (WP1) and the second peripheral wall portion (WP2) to form a continuous surface, corresponding to the straight portion of the oval.

8. The observation holder (2) according to claim 5, wherein: a positioning portion (210a) for positioning the first plate material (221) is provided on the lower surface of the housing portion main body (210).

9. The observation holder (2) according to claim 4, wherein: the housing portion main body (210) and the housing portion upper part (211) are configured such that the widths of the housing portion main body (210) and the housing portion upper part (211) in one horizontal direction are equal to each other.

10. The observation holder (2) according to claim 1, wherein: it further includes an information providing portion (23) that can provide information about the observation object.

11. An observation holder (2) includes: a housing portion (21) configured to house an observation object, and an observation portion (22) provided at the lower part of the housing portion (21), wherein: a plurality of holes (H1) for housing the observation object are provided in the housing portion (21) so as to extend in the vertical direction and be arranged side by side in the horizontal direction; the observation portion (22) includes: a first plate material (221) that covers the housing portion (21) from below; and a second plate material (222) having translucency that covers the first plate material (221) from below; a plurality of through holes (H3) corresponding to the plurality of holes (H1) are provided in the first plate material (221); a concave portion (CC1) that connects two adjacent through holes among the plurality of through holes (H3) is provided on the lower surface of the first plate material (221), and the outer edge of the concave portion (CC1) surrounds the two through holes; the housing portion (21) of the observation holder (2) has a rectangular shape when viewed from above.

12. An observation device (1), characterized in that, It includes: a support table (11) that supports a plurality of the observation holders (2) according to claim 1 in parallel and arranges the plurality of observation holders (2) in the horizontal direction; a photographing portion (12) for photographing the observation objects housed in the plurality of observation holders (2); and a moving mechanism (13) that relatively moves the photographing portion (12) relative to the support table (11) in the horizontal direction.

13. The observation device (1) according to claim 12, characterized in that: it further comprises an observation preparation unit (14) for accommodating an object to be observed in the aforementioned observation holder (2), the aforementioned observation preparation unit (14) includes: a preparation support table (141) for supporting the aforementioned observation holder (2) in a state before accommodating the object to be observed, and a second moving mechanism (142) capable of moving the aforementioned preparation support table (141) in a specified direction.

14. An observation wafer that can be mounted on an observation holder (2), the observation holder (2) having: a housing part (21) configured to be able to accommodate an object to be observed, and an observation part (22) provided at the lower part of the housing part (21), the aforementioned observation wafer being a component of the aforementioned observation part (22), characterized in that: a plurality of holes (H1) for accommodating an object to be observed are provided in the aforementioned housing part (21) so as to extend in the vertical direction and be arranged side by side in the horizontal direction, the aforementioned observation wafer includes: a first plate (221) covering the aforementioned housing part (21) from below; and a second plate (222) having light transmissivity and covering the aforementioned first plate (221) from below, a plurality of through holes (H3) corresponding to the aforementioned plurality of holes (H1) are provided in the aforementioned first plate (221), a recess (CC1) connecting two adjacent through holes among the aforementioned plurality of through holes (H3) is provided on the lower surface of the aforementioned first plate (221), and the outer edge of the recess (CC1) surrounds the two through holes, the aforementioned recess (CC1) is provided to be oval-shaped in a top view, the side wall (WP) forming the aforementioned recess (CC1) includes: a first peripheral wall part (WP1) provided along a part of the outer edge of one of the aforementioned adjacent two through holes, corresponding to the circumferential part in the aforementioned oval; a second peripheral wall part (WP2) provided along a part of the outer edge of the other of the aforementioned adjacent two through holes, corresponding to the other circumferential part in the aforementioned oval; and a third peripheral wall part (WP3) connecting the aforementioned first peripheral wall part (WP1) and the aforementioned second peripheral wall part (WP2) to form a continuous surface, corresponding to the straight part in the aforementioned oval.

15. A method for manufacturing an observation wafer, which is the method for manufacturing the observation wafer according to claim 14, characterized in that: it includes: a step of overlapping the aforementioned second plate (222) on the lower surface of the aforementioned first plate (221); and a step of bonding the aforementioned first plate (221) and the aforementioned second plate (222) by anodic bonding.

Images