WO2022247944A1 - 一种计数组件及计数装置及应用 - Google Patents

一种计数组件及计数装置及应用 Download PDF

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Publication number
WO2022247944A1
WO2022247944A1 PCT/CN2022/095775 CN2022095775W WO2022247944A1 WO 2022247944 A1 WO2022247944 A1 WO 2022247944A1 CN 2022095775 W CN2022095775 W CN 2022095775W WO 2022247944 A1 WO2022247944 A1 WO 2022247944A1
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Prior art keywords
counting
sample
main body
counting assembly
assembly according
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PCT/CN2022/095775
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English (en)
French (fr)
Inventor
陈凯
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上海睿钰生物科技有限公司
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Publication of WO2022247944A1 publication Critical patent/WO2022247944A1/zh
Priority to US18/518,934 priority Critical patent/US20240084239A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles

Definitions

  • This specification relates to the technical field of biomedical instruments, in particular to a counting component, counting device and application.
  • the counting plate is a common consumable for the identification and counting of microparticles and cells, and is widely used in the fields of life sciences, biomedicine, and medical testing. At present, most of the counting panels used in cell counters at home and abroad are realized by referring to the counting principle of the hemocytometer. Taking the classic hemocytometer as an example, only part of the limited volume (random sampling) in the counting pool is imaged and counted, and the full volume imaging count cannot be realized. Inhomogeneous distribution of cells in the counting area will generate inherent distribution errors, and when the cell concentration is low, this distribution error will be significantly increased, greatly reducing the counting accuracy. Therefore, there is an urgent need for a counting plate that can realize imaging of the entire sample volume, improve counting accuracy, and expand the countable range.
  • One of the embodiments of this specification provides a counting assembly, including a main body, on which a concave sample groove is provided, the bottom end of the sample groove is a plane, and the cross-sectional area of the sample groove from the bottom end to the top is Gradually increase.
  • the sample groove is in the shape of a rounded truncated, oblong truncated or chamfered truncated edge.
  • the angle between the inner wall of the sample tank and the horizontal plane is 45-60°.
  • the body is a thin-walled structure.
  • the thin-walled structure has a uniform wall thickness.
  • the main body is a thin-walled structure with an open bottom.
  • the body is a hollow, thin-walled closed structure, or a solid structure.
  • the hollow thin-walled closed structure generally refers to the hollow between the outer wall of the main body and the side wall of the sample tank.
  • the solid structure generally refers to the solid filling between the outer wall of the main body and the side wall of the sample tank.
  • a cover plate is also included, the upper surface of the main body is attached to the lower surface of the cover plate, and a sample injection hole and an exhaust hole are opened on the cover plate.
  • the projections of the sample injection hole and the exhaust hole on the horizontal plane do not overlap with the projection of the bottom surface of the sample tank on the horizontal plane.
  • the area of the lower surface of the cover plate is larger than the area of the upper surface of the main body.
  • the main body can be completely covered when the cover plate is placed on the main body.
  • the shape of the lower surface of the cover plate may be similar to that of the upper surface of the main body.
  • both the lower surface of the cover plate and the upper surface of the main body may be circular, square, rectangular or the like.
  • the shape of the lower surface of the cover plate may be different from the shape of the upper surface of the main body.
  • the lower surface of the cover plate may be square, while the upper surface of the main body may be circular.
  • the shape of the exhaust hole and/or the sample injection hole is circular, oval, rectangular, rhombus or irregular.
  • a first protrusion is provided on the upward side of the edge of the sample injection hole and/or the exhaust hole.
  • the first protrusion faces the opposite side of the main body, and is used to prevent sample overflow caused by adding a slightly excessive amount of sample, so as to improve the error tolerance rate of the operation.
  • a second protrusion is provided on the downward side of the edge of the sample injection hole and/or the exhaust hole, and the outer side of the second protrusion is in contact with the inner wall of the sample groove.
  • the bottom surface of the sample well is smaller than the imaging target surface of the camera, so as to ensure that a single imaging field of view can be completely covered.
  • the imaging target surface of the camera Those skilled in the art know the area range of the imaging target surface of the camera.
  • the diameter of the bottom surface of the sample well is 1.5-2.0 mm.
  • One of the embodiments of the present specification provides a counting device, including the counting component described in any embodiment of the present application.
  • the counting component includes a cover plate, and the counting device further includes a carrier plate; the carrier plate is provided with at least one hole matching the counting component, and the diameter of the hole is within the diameter of the main body. between the outer diameter of the cover plate and the diameter of the cover plate, so that the main body can be inserted into the hole and the cover plate can be stuck outside the hole.
  • 24, 48 or 96 holes are opened on the carrier plate, and the counting component is fixedly installed in each of the holes.
  • the carrier board is integrally formed with the counting component, or is detachably connected.
  • the detachable connection may rely on the friction force between the outer wall of the main body and the inner wall of the hole, so that the main body is snapped into the hole.
  • the hole can also be set in a corresponding shape.
  • the edge of the support plate is provided with a support edge, and the height of the support edge extending toward the bottom side of the counting assembly is greater than the height of the counting assembly.
  • the counting component can be the counting component as described in any embodiment of the present application; the counting device can be The counting device as described in any embodiment of the present application.
  • the imaging and counting of cells, bacteria, fungi or other particles in the full volume of the sample in the sample tank for example, 5 ⁇ L, 20 ⁇ L, 100 ⁇ L, 200 ⁇ L, 300 ⁇ L, 400 ⁇ L, 500 ⁇ L), especially suitable for low concentrations (down to 0 Count detection of cells, bacteria, fungi or other particles in samples/ml).
  • Fig. 1 is a schematic structural diagram of a counting component according to some embodiments of the present specification
  • Fig. 2 is a schematic top view of a counting assembly according to some embodiments of the present specification
  • Fig. 3 is a schematic structural view of a carrier plate of a counting device according to some embodiments of the present specification
  • Fig. 4 is a schematic structural diagram of a counting device according to some embodiments of the present specification.
  • Fig. 5 is a schematic structural diagram of a counting device according to other embodiments of the present specification.
  • Fig. 6 is a schematic cross-sectional structural view of a counting device according to other embodiments of the present specification.
  • the embodiment of the present application relates to a counting component.
  • the counting component can enrich the cells, bacteria, fungi or other particles in the sample in the sample tank at the bottom, and realize the full volume of the sample in the sample tank (for example, 5 ⁇ L, 20 ⁇ L, 100 ⁇ L, 200 ⁇ L, 300 ⁇ L, 400 ⁇ L, 500 ⁇ L) imaging and counting of cells, bacteria, fungi or other particles, especially suitable for the counting and detection of cells, bacteria, fungi or other particles in low-concentration samples.
  • Fig. 1 is a schematic structural view of a counting assembly according to some embodiments of this specification
  • Fig. 2 is a schematic top view of a counting assembly according to some embodiments of this specification.
  • the counting component involved in the embodiment of the present application will be described in detail below with reference to FIG. 1 and FIG. 2 . It should be noted that the following examples are only used to explain the present application, and do not constitute a limitation to the present application.
  • the counting assembly 100 includes a main body 110 and a cover plate 120, the main body 110 is provided with a concave sample groove (as shown by the dotted line in the main body 110 in Figure 1), and the bottom end of the sample groove is a plane , the cross-sectional area of the sample tank gradually increases from the bottom to the top.
  • the objects to be counted in the sample can be concentrated on the bottom surface of the sample tank due to the centrifugal force.
  • the objects to be counted in the sample can also be concentrated on the bottom surface of the sample groove by means of standing still or magnetic attraction.
  • the counting can be observed and counted through a microscope or counted and analyzed after taking pictures with an automatic counting instrument. Wherein, when the imaging field of view of the camera of the automatic counter can completely cover the plane at the bottom of the main body 110, the imaging counting analysis can be realized through one imaging.
  • the imaging counting analysis can be performed by splicing the pictures after taking multiple shots.
  • the bottom surface of the sample well is circular, and the diameter of the circle is 1.5-2.0 mm, which is smaller than the actual imaging target surface of the camera, so as to ensure that the single imaging field of view can be completely covered.
  • those skilled in the art can select a centrifuge according to the size of the main body 110, or design the size of the main body 110 according to the specifications of the existing centrifuge.
  • the sample well is in the shape of a rounded frustum.
  • the sample groove is in the shape of an inverted ellipse.
  • the sample well is in the shape of a chamfered truncated edge.
  • the sample groove is rounded truncated (for example, the inner diameter of the top is 9mm, the depth is 3.7mm, and the inner diameter of the bottom is 2mm), and the sample is placed inside the rounded truncated table, After centrifugation, the object to be detected can be enriched in the bottom plane of the rounded table in the sample without accumulation on the side wall.
  • the sample groove By setting the sample groove into a regular shape (such as a rounded truncated shape), the sample groove can be easily processed and manufactured.
  • the sample wells can be of other shapes.
  • the sample groove can be in any shape that satisfies "the bottom end is flat, and the cross-sectional area gradually increases from the bottom end to the top end".
  • the angle between the inner wall of the sample tank and the horizontal plane is 45-60° (such as 45°, 50°, 60°, etc.), and the inwardly inclined sidewall is conducive to the sinking of the detected object to enrich on the bottom of the sample well.
  • the angle between the inner wall of the sample tank and the horizontal plane should not be too small. If the angle is too small, it will make it difficult for the object to be detected to be completely concentrated at the bottom of the sample tank. In some embodiments, the angle between the inner wall of the sample chamber and the horizontal plane should not be too large (eg close to 90°), otherwise shadows will be generated during imaging and counting.
  • the main body 110 may be of a thin-walled structure, so as to facilitate manufacturing (eg, not easily deformed during the manufacturing process) and save manufacturing cost.
  • the thin-walled body 110 has a uniform wall thickness.
  • the main body 110 may be a thin-walled structure with an open bottom.
  • the main body 110 may be a hollow thin-walled closed structure, and the outer wall of the main body 110 and the side wall of the sample chamber are hollow.
  • the main body 110 may also be a solid structure, that is, the space between the outer wall of the main body 110 and the side wall of the sample chamber is filled with solid.
  • the counting assembly 100 further includes a cover plate 120, the upper surface of the main body 110 is attached to the lower surface of the cover plate 120, and the cover plate 120 covers the main body 110 during use, and the gap between the main body 110 and the cover plate 120 is
  • the fixed connection can be realized by bonding, clamping or integral molding, or the two can only be in contact without being fixedly connected.
  • the cover plate 120 is respectively provided with an exhaust hole 122 for exhaust and a sample injection hole 121 for adding a sample.
  • the exhaust hole 122 and the sample injection hole 121 can be the same shape or different shapes. 122 or the shape of the sample injection hole 121 can be circular, oval, rectangular, rhombus, or irregular.
  • the area of the lower surface of the cover plate 120 may be greater than the area of the upper surface of the main body 110 , so that the cover plate 120 can completely cover the main body 110 when placed on the main body 110 .
  • the shape of the lower surface of the cover plate 120 may be similar to the shape of the upper surface of the main body 110 .
  • both the lower surface of the cover plate 120 and the upper surface of the main body 110 may be circular, square, or rectangular.
  • the shape of the lower surface of the cover plate 120 may be different from the shape of the upper surface of the main body 110 .
  • the lower surface of the cover plate 120 may be square, while the upper surface of the main body 110 may be circular. In the embodiment shown in FIGS.
  • the outer contours of the cover plate 120 and the main body 110 are both cylindrical.
  • the diameter of the cover plate 120 may be larger than that of the main body 110 .
  • the outer diameter of the cylindrical body 110 is 6mm, 7mm, 8mm, 9mm or 10mm, and the diameter of the circular cover 120 is 11.5mm or 12mm.
  • the projection of the sample injection hole 121 and the exhaust hole 122 on the horizontal plane does not overlap with the projection of the bottom surface of the sample tank on the horizontal plane, thereby avoiding the need for the sample injection hole 121 or the exhaust hole during microscope observation or imaging photography.
  • the profile structure of 122 affects the count.
  • the elliptical sample injection hole 121 for example, the inner diameter of the major axis is 3mm, and the inner diameter of the minor axis is 1.5mm
  • the rectangular exhaust hole 122 for example, long 3mm, wide 1mm
  • the sample injection hole 121 and the exhaust hole 122 can also be set at other near edge positions on the cover plate 120, and the bottom end plane can also be set directly below the center position of the cover plate 120, so that adding The projection of the sample hole 121 and the exhaust hole 122 on the horizontal plane does not overlap with the projection of the bottom surface of the sample tank on the horizontal plane.
  • a first protrusion is provided on the upward side of the edge of the sample injection hole 121 and the exhaust hole 122. As shown in FIG. The sample overflow caused by a slight excess increases the error tolerance rate of the operation. In some other embodiments, only the first protrusion is provided on the upward side of the edge of the sample injection hole 121 or the first protrusion is provided on the upward side of the exhaust hole 122, which can also prevent the sample from overflowing to a certain extent. effect.
  • a second protrusion is provided on the downward side of the edge of the sample injection hole 121 and the exhaust hole 122 , and the outer side of the second protrusion is in contact with the inner wall of the sample groove.
  • the second protrusion can be used for contact between the main body 110 and the cover plate 120 without a fixed connection. Carry out the function of limit and stop, so that the cover plate 120 does not shift after the cover plate 120 is covered, and also ensure that the sample injection hole 121 and the exhaust hole 122 are communicated with the inside of the sample tank, thereby ensuring the effect of sample addition and exhaust .
  • the positioning of the main body 110 and the cover plate 120 during fixed installation can be facilitated by providing the second protrusion.
  • only the edge of the sampling hole 121 facing downward is provided with a second protrusion or only the side of the exhaust hole 122 facing downward is provided with a second protrusion, and the outer side of the second protrusion is in contact with the side of the sample groove.
  • the contact of the inner wall can also play a certain position-limiting effect on the cover plate 120 .
  • the embodiment of the present application relates to a counting device.
  • the counting device includes the counting assembly 100 of any of the above-mentioned technical schemes, through which the cells, bacteria, fungi or other particles in the sample are enriched at the bottom, and by directly observing or imaging the bottom plane, it is possible to monitor the sample tank. Imaging counting of cells, bacteria, fungi or other particles in the sample volume, especially suitable for the counting and detection of cells, bacteria, fungi or other particles in low-concentration samples.
  • Fig. 3 is a schematic structural view of a carrier plate of a counting device according to some embodiments of the present specification.
  • the counting device can also include a carrier plate 200; the carrier plate 200 is provided with at least one hole 210 matching the counting assembly 100, so that the main body 110 can be inserted into the hole 210 and the cover plate 120 Can be stuck out of hole 210 .
  • the diameter of the hole 210 is 11 mm
  • the outer diameter of the cylindrical body 110 is 10.5 mm
  • the diameter of the circular cover 120 is 12 mm.
  • the diameter of the hole 210 is 10.5 mm
  • the outer diameter of the cylindrical body 110 is 9.83 mm
  • the diameter of the cover plate 120 is 11.5 mm.
  • Fig. 4 is a schematic structural diagram of a counting device according to some embodiments of the present specification.
  • a small number of holes 210 (for example, 2 holes, 3 holes, 4 holes, 5 holes) can be opened on the carrier board 200, and the main body 110 can be movably inserted into the holes 210 and connected with the carrier board. 200 detachable connections.
  • the detachable connection can be made by setting the diameter of the hole 210 between the outer diameter of the main body 110 and the diameter of the cover plate 120, wherein the main body 110 is fixedly connected with the cover plate 120, and the diameter of the cover plate 120 is larger than the outer diameter of the main body 110 (For example, the diameter of the hole 210 is 10.5 mm, the maximum outer diameter of the main body 110 is 9.83 mm, and the diameter of the cover plate 120 is 11.5 mm).
  • the detachable connection may rely on the friction force between the outer wall of the main body 110 and the inner wall of the hole 210 , so that the main body 110 snaps into the hole 210 .
  • the hole 210 when the outer contour of the main body 110 is a different shape (such as a cuboid), the hole 210 can also be set in a corresponding shape.
  • the single counting assembly 100 can be separated from the carrier plate 200, and the size of the single counting assembly 100 can be matched with the existing centrifuge device, so it can be placed in the centrifuge device separately for centrifugation, so that the detected substance Sink to the bottom. Then put it into the hole 210 of the carrier plate 200, and use a microscope or an automatic counter for counting analysis.
  • Fig. 5 is a schematic structural view of a counting device according to other embodiments of the present specification
  • Fig. 6 is a schematic cross-sectional structural view of a counting device according to other embodiments of the present specification.
  • a plurality of holes 210 can be opened on the carrier plate 200, and a counting assembly 100 is fixedly installed in each hole 210. , forming a high-throughput counting device.
  • the fixed connection method can be bonding. Bonding can be done with UV glue or glue injection. For example, the edge of the lower surface of the cover plate 120 of the counting assembly 100 and the carrier plate 200 can be glued together.
  • the carrier plate 200 and the counting assembly 100 can be integrally formed to form an integrated counting device with the same dimensions and specifications as the existing 24-well plate, 48-well plate or 96-well plate.
  • the counting device can be matched with the centrifuge device, so it can be put into the centrifuge device as a whole for centrifugation, so that the substance to be detected sinks to the bottom. Then count and analyze using an automatic counter.
  • a support edge 220 may be provided on the edge of the carrier plate 200 , and the height of the support edge 220 extending toward the bottom side of the counting assembly 100 is greater than the height of the counting assembly.
  • the supporting edge 220 can be arranged on any two opposite sides of the carrier plate 200, and can also be arranged on the four sides of the carrier plate 200. When the carrier plate 200 is placed, the bottom of the supporting edge 220 can be directly in contact with the desktop, table, etc., so as to avoid the bottom of the counting assembly 100. Direct contact with desktops, countertops, etc. will be scratched, which will affect the counting analysis.
  • the material of the counting device may be a material with high transparency and chemical corrosion resistance.
  • a material with high transparency and chemical corrosion resistance for example, polypropylene, polystyrene, and the like.
  • the possible beneficial effects of the embodiments of the present application include but are not limited to: (1)
  • the counting device can be washable and reusable, or it can be disposable, and it is easy to operate when used as a single use, and does not need to be installed and cleaned; (2) )
  • the imaging and counting of the full volume of particles or cells in the sample tank can be realized by at least one imaging, the detection is fast and efficient, and the counting error caused by the distribution of particles or cells and/or the limited sampling problem is solved and eliminated; (3)
  • the single detection sample volume can reach 500 ⁇ L, and the lower limit range of sample concentration detection is greatly expanded, the lowest can reach 0/ml, which is especially suitable for the counting detection of low concentration samples.
  • this application can adjust the size of counting components and counting devices according to application requirements, so that it can be used not only for microparticle counting, but also for accurate counting and fluorescence analysis of various cells, as well as blood cell counting in medical clinical analysis. , urine formed component analysis and other fields. It should be noted that different embodiments may have different beneficial effects, and in different embodiments, the possible beneficial effects may be any one or a combination of the above, or any other possible beneficial effects.
  • numbers describing the quantity of components and attributes are used. It should be understood that such numbers used in the description of the embodiments use the modifiers "about”, “approximately” or “substantially” in some examples. grooming. Unless otherwise stated, “about”, “approximately” or “substantially” indicates that the stated figure allows for a variation of ⁇ 20%. Accordingly, in some embodiments, the numerical parameters used in the specification and claims are approximations that can vary depending upon the desired characteristics of individual embodiments. In some embodiments, numerical parameters shall take into account the specified significant digits and adopt the general digit reservation method. Although the numerical ranges and parameters used in some embodiments of this specification to confirm the breadth of the range are approximations, in specific embodiments, such numerical values are set as precisely as practicable.

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Abstract

一种计数组件（100）及计数装置及应用。计数组件（100）包括主体（110），主体（110）上设有内凹的样品槽，样品槽的底端为平面，样品槽由底端至顶端的横截面面积逐渐增大。使用计数组件（100）能通过最少一次成像即可实现样品槽内样品全体积颗粒或细胞等的成像计数，检测快速高效，解决并消除了由于颗粒或细胞等的分布问题和/或采样有限问题而产生的计数误差。

Description

一种计数组件及计数装置及应用
交叉引用
本申请要求2021年5月28日提交的申请号为202121186301.X的中国申请优先权，其全部内容通过引用的方式被包含于此。
技术领域
本说明书涉及生物医学仪器技术领域，特别涉及一种计数组件及计数装置及应用。
背景技术
计数板是一种用于微颗粒和细胞识别、计数的常用耗材，广泛应用于生命科学、生物医药和医学检验等领域。目前国内外细胞计数仪所用计数板，大多是参照血球计数板的计数原理实现的。以经典的血球计数板为例，只对计数池内部分有限体积(随机取样)的成像计数，无法实现全体积成像计数。细胞在计数区域分布不均匀会产生固有的分布误差，当细胞浓度较低时，还会显著增大此分布误差，极大地降低计数精确度。因此亟需一种可实现全样品体积成像的计数板，提高计数精确度，拓展可计数范围。
发明内容
本说明书实施例之一提供一种计数组件，包括主体，所述主体上设有内凹的样品槽，所述样品槽的底端为平面，所述样品槽由底端至顶端的横截面面积逐渐增大。
在一些实施例中，所述样品槽为倒圆台形、倒椭圆台形或倒棱台形。
在一些实施例中，所述样品槽的内壁与水平面的夹角为45-60°。
在一些实施例中，所述主体为薄壁结构。
在一些实施例中，所述薄壁结构的壁厚均一。
在一些实施例中，所述主体为下部敞开的薄壁结构。
在一些实施例中，所述主体为空心的薄壁封闭结构，或者实心结构。其中，空心的薄壁封闭结构一般是指主体的外壁与样品槽的侧壁之间空心。实心结构一般是指主体的外壁与样品槽的侧壁之间为实心填充。
在一些实施例中，还包括盖板，所述主体的上表面与所述盖板的下表面相贴合，所述盖板上开设有加样孔和排气孔。
在一些实施例中，所述加样孔和所述排气孔在水平面上的投影与样品槽的底面在水平面上的投影不重叠。
在一些实施例中，所述盖板的下表面的面积大于所述主体的上表面的面积。以使得所述盖板盖于所述主体上方时能够完全覆盖主体。所述盖板的下表面的形状可以和所述主体的 上表面的形状相似。例如，所述盖板的下表面的和所述主体的上表面可以均为圆形、正方形、矩形等。在一些实施例中，所述盖板的下表面的形状可以和所述主体的上表面的形状不同。例如，所述盖板的下表面可以为正方形，而主体的上表面可以为圆形。
在一些实施例中，所述排气孔和/或所述加样孔的形状为圆形、椭圆形、矩形、菱形或不规则形状。
在一些实施例中，所述加样孔和/或所述排气孔的边缘朝上一侧设置有第一凸起。所述第一凸起朝向所述主体的相对一侧，用于防止在加样稍微过量造成的样品溢出，提高操作的容错率。
在一些实施例中，所述加样孔和/或所述排气孔的边缘朝下一侧设置有第二凸起，所述第二凸起外侧与样品槽的内壁接触。
在一些实施例中，所述样品槽的底面小于相机的成像靶面，保证单次成像视野可完全覆盖。本领域技术人员均知晓相机的成像靶面的面积范围。
在一些实施例中，所述样品槽的底面直径为1.5-2.0mm。
本说明书实施例之一提供一种计数装置，包括如本申请任一实施例所述的计数组件。
在一些实施例中，所述计数组件包括盖板，所述计数装置还包括载板；所述载板上开设有至少一个与所述计数组件匹配的孔洞，所述孔洞的直径在所述主体的外径与所述盖板的直径之间，使得所述主体能够插入所述孔洞中且所述盖板能够被卡在所述孔洞外。
在一些实施例中，所述载板上开设有24、48或96个孔洞，每个所述孔洞中固定安装有所述计数组件。
在一些实施例中，所述载板与所述计数组件一体成型，或者可拆分连接。所述可拆分连接可以为依靠所述主体外壁与所述孔洞的内壁的摩擦力，使得所述主体卡入所述孔洞中。在一些实施例中，当所述主体的外轮廓为不同形状(如长方体)时，所述孔洞也可以设置为对应形状。
在一些实施例中，所述载板边缘设置有支撑边，所述支撑边向计数组件的底部一侧延伸的高度大于所述计数组件的高度。
本说明书实施例之一提供一种计数组件和/或计数装置在微粒计数分析领域中的应用，所述计数组件可为如本申请任一实施例所述的计数组件；所述计数装置可为如本申请任一实施例所述的计数装置。通过该应用，可实现对样品槽内样品全体积(例如，5μL、20μL、100μL、200μL、300μL、400μL、500μL)细胞、细菌、真菌或其他微粒成像计数，尤其适合低浓度(最低可达0个/ml)样品中细胞、细菌、真菌或其他微粒的计数检测。
附图说明
本说明书将以示例性实施例的方式进一步说明，这些示例性实施例将通过附图进行详细描述。这些实施例并非限制性的，在这些实施例中，相同的编号表示相同的结构，其中：
图1是根据本说明书一些实施例所示的计数组件的结构示意图；
图2是根据本说明书一些实施例所示的计数组件的俯视结构示意图；
图3是根据本说明书一些实施例所示的计数装置的载板的结构示意图；
图4是根据本说明书一些实施例所示的计数装置的结构示意图；
图5是根据本说明书另一些实施例所示的计数装置的结构示意图；
图6是根据本说明书另一些实施例所示的计数装置的剖面结构示意图；
附图标记说明：100-计数组件，110-主体，120-盖板，121-加样孔，122-排气孔，200-载板，210-孔洞，220-支撑边。
具体实施方式
为了更清楚地说明本说明书实施例的技术方案，下面将对实施例描述中所需要使用的附图作简单的介绍。显而易见地，下面描述中的附图仅仅是本说明书的一些示例或实施例，对于本领域的普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图将本说明书应用于其它类似情景。除非从语言环境中显而易见或另做说明，图中相同标号代表相同结构或操作。如本说明书和权利要求书中所示，除非上下文明确提示例外情形，“一”、“一个”、“一种”和/或“该”等词并非特指单数，也可包括复数。
本申请实施例涉及一种计数组件。该计数组件能够将样品槽内样品中的细胞、细菌、真菌或其他微粒富集在底部，通过对底部平面直接观察或成像，实现对样品槽内样品全体积(例如，5μL、20μL、100μL、200μL、300μL、400μL、500μL)细胞、细菌、真菌或其他微粒成像计数，尤其适合低浓度样品中细胞、细菌、真菌或其他微粒的计数检测。
图1是根据本说明书一些实施例所示的计数组件的结构示意图；图2是根据本说明书一些实施例所示的计数组件的俯视结构示意图。以下将结合图1、图2对本申请实施例所涉及的计数组件进行详细说明。值得注意的是，以下实施例仅仅用以解释本申请，并不构成对本申请的限定。
如图1和图2所示，计数组件100包括主体110和盖板120，主体110上设有内凹的样品槽(如图1中主体110内的虚线所示)，样品槽底端为平面，样品槽由底端至顶端的横截面面积逐渐增大。通过形成顶端横截面大于底端横截面且侧面均匀平缓过渡的结构，样品槽内能够置入较大体积的液体。通过将带有样品的主体离心，能够使得样品中待计数的对象(如细胞、细菌、真菌或其他微粒等)由于离心力的作用被富集于样品槽的底面上。在一些 实施例中，也可以通过静置、磁吸等方式使得样品中的待计数对象富集到样品槽的底面上。待计数对象富集完成后，可以通过显微镜进行观察计数或通过自动计数仪拍摄图片后计数分析。其中，自动计数仪的相机成像视野能够完全覆盖主体110底端的平面时，通过一次成像即可实现成像计数分析。自动计数仪的相机成像视野不能够完全覆盖主体110底端平面时，可通过多次拍摄再将图片拼接后进行成像计数分析。在一些实施例中，样品槽底面为圆形，该圆形的直径为1.5-2.0mm，小于相机的实际成像靶面，保证单次成像视野可完全覆盖。在一些实施例中，本领域技术人员可以根据主体110的尺寸选择离心机，也可根据现有离心机的规格设计主体110的尺寸。
在一些实施例中，样品槽为倒圆台形。在另一些实施例中，样品槽为倒椭圆台形。在又一些实施例中，样品槽为倒棱台形。在一些实施例中，如图1和图2所示，样品槽为倒圆台形(例如，顶端内径为9mm，深度为3.7mm，底端内径为2mm)，样品置入后位于倒圆台内部，离心后样品中实现待检测对象在倒圆台底端平面富集，而不会在侧壁堆积。通过将样品槽设置为规则形状(如倒圆台形)，能够使得样品槽便于加工制造。在一些实施例中，样品槽可以为其他形状。例如，样品槽可以为满足“底端为平面，由底端至顶端的横截面面积逐渐增大”的任意形状。
在一些实施例中，样品槽的内壁与水平面的夹角为45-60°(如45°、50°、60°等)，向内倾斜的侧壁有利于被检测对象沉底，以富集在样品槽底面上。在一些实施例中，样品槽的内壁与水平面的夹角不宜过小，若夹角过小则会造成待检测对象难以完全富集在样品槽底部。在一些实施例中，样品槽的内壁与水平面的夹角不宜过大(如接近90°)，若夹角过大则会造成成像计数的时候产生阴影。
在一些实施例中，主体110可以为薄壁结构，以便于生产制造(如生产过程中不易变形)且能够节约制造成本。在一些实施例中，薄壁结构的主体110的壁厚均一。在一些实施例中，主体110可以为下部敞开的薄壁结构。在一些实施例中，主体110可以为空心的薄壁封闭结构，主体110的外壁与样品槽的侧壁之间空心。在另一些实施例中，主体110也可以为实心结构，即主体110的外壁与样品槽的侧壁之间为实心填充。
在一些实施例中，计数组件100还包括盖板120，主体110的上表面与盖板120的下表面相贴合，使用时盖板120覆盖在主体110上，主体110与盖板120之间可以通过粘接、卡接或一体成型的方式实现固定连接，也可以仅仅两者相接触而不固定连接。盖板120上分别设置有用于排气的排气孔122和用于加样的加样孔121，排气孔122和加样孔121可以为相同的形状也可以为不同的形状，排气孔122或加样孔121的形状可以为圆形、椭圆形、矩形、菱形或不规则形状等。
在一些实施例中，盖板120下表面的面积可以大于主体110上表面的面积，以使得盖板120盖于主体110上方时能够完全覆盖主体110。在一些实施例中，盖板120下表面的形状可以和主体110上表面的形状相似。例如，盖板120下表面的和主体110上表面可以均为圆形、正方形、矩形等。在一些实施例中，盖板120下表面的形状可以和主体110上表面的形状不同。例如，盖板120下表面可以为正方形，而主体110上表面可以为圆形。在图1-2所示的实施例中，盖板120和主体110外轮廓均为圆柱体。盖板120的直径可以大于主体110的直径。例如，柱形主体110的外径为6mm、7mm、8mm、9mm或10mm，圆形盖板120的直径为11.5mm或12mm。
在一些实施例中，加样孔121和排气孔122在水平面上的投影与样品槽的底面在水平面上的投影不重叠，从而能够避免显微镜观察或成像摄影时加样孔121或排气孔122的轮廓结构对计数造成影响。如图2所示，椭圆形加样孔121(例如，长轴内直径为3mm，短轴内直径为1.5mm)和矩形排气孔122(例如，长3mm，宽1mm)沿盖板120的径向对称设置在盖板120的近边缘处，样品槽底端平面设置在盖板120中心位置的正下方。在另一些实施例中，加样孔121和排气孔122也可以设置在盖板120上的其他近边缘位置，底端平面也可以设置在靠近盖板120中心位置的正下方处，使得加样孔121和排气孔122在水平面的投影与样品槽底面在水平面的投影不重叠。
在一些实施例中，加样孔121和排气孔122边缘朝上一侧设置有第一凸起，如图1所示，该凸起朝向主体110的相对一侧，用于防止在加样稍微过量造成的样品溢出，提高操作的容错率。在另一些实施例中，仅加样孔121边缘朝上一侧设置有第一凸起或排气孔122边缘朝上一侧设置有第一凸起，也可以起到一定程度的防样品溢出作用。
在一些实施例中，加样孔121和排气孔122边缘朝下一侧设置有第二凸起，第二凸起外侧与样品槽的内壁接触。在一些实施例中，第二凸起可以用于主体110和盖板120之间相接触而无固定连接的情况，在盖上盖板120时，该第二凸起能够起到对盖板120进行限位和卡位的作用，使得盖上盖板120后盖板120不发生偏移，也保证加样孔121和排气孔122与样品槽内部联通，从而保证加样和排气的效果。在一些实施例中，通过设置第二凸起可以便于主体110和盖板120在固定安装时定位。在另一些实施例中，仅加样孔121边缘朝下一侧设置有第二凸起或仅排气孔122边缘朝下一侧设置有第二凸起，第二凸起外侧与样品槽的内壁接触，也可以对盖板120起到一定的限位作用。
本申请实施例涉及一种计数装置。该计数装置包括上述任一技术方案的计数组件100，通过计数组件100将样品中的细胞、细菌、真菌或其他微粒等富集在底部，通过对底部平面直接观察或成像，实现对样品槽内样品全体积细胞、细菌、真菌或其他微粒等的成像计数， 尤其适合低浓度样品中细胞、细菌、真菌或其他微粒等的计数检测。
图3是根据本说明书一些实施例所示的计数装置的载板的结构示意图。如图3所示，在一些实施例中，计数装置还可以包括载板200；载板200上开设有至少一个与计数组件100匹配的孔洞210，使得主体110能够插入孔洞210中且盖板120能够被卡在孔洞210外。例如，孔洞210的直径为11mm，柱形主体110的外径为10.5mm，圆形盖板120的直径为12mm。又例如，孔洞210的直径为10.5mm，柱形主体110的外径为9.83mm，盖板120的直径为11.5mm。
图4是根据本说明书一些实施例所示的计数装置的结构示意图。如图4所示，在一些实施例中，载板200上可以开设有少量孔洞210(例如，2孔、3孔、4孔、5孔)，主体110能够活动插入孔洞210中并与载板200可拆分连接。在一些实施例中，可拆分连接可以为通过设置孔洞210的直径在主体110外径和盖板120直径之间，其中主体110与盖板120固定连接，盖板120直径大于主体110外径(例如，孔洞210直径10.5mm，主体110最大外径9.83mm，盖板120直径11.5mm)，将主体110插入孔洞210中，通过盖板120架设在载板200上。在一些实施例中，可拆分连接可以为依靠主体110外壁与孔洞210内壁的摩擦力，使得主体110卡入孔洞210中。在一些实施例中，当主体110外轮廓为不同形状(如长方体)时，孔洞210也可以设置为对应形状。
本实施例中，单个的计数组件100能够同载板200分离，单个的计数组件100的尺寸能够与现有的离心装置匹配，因此能够将其单独置入离心装置中进行离心，使得待检测物沉底。然后将其置入载板200的孔洞210中，利用显微镜或自动计数仪进行计数分析。
图5是根据本说明书另一些实施例所示的计数装置的结构示意图；图6是根据本说明书另一些实施例所示的计数装置的剖面结构示意图。如图5和图6所示，在一些实施例中，载板200上可以开设有多个孔洞210(例如，24孔、48孔、96孔)，每个孔洞210中固定安装有计数组件100，形成高通量的计数装置。固定连接方式可以为粘接。粘接可以采用UV胶粘合或注胶液粘合。例如，可以将计数组件100的盖板120的下表面边缘与载板200通过胶液粘合。在一些实施例中，载板200与计数组件100可以一体成型，以形成与现有的24孔板、48孔板或96孔板相同尺寸规格的一体化计数装置。在一些实施例中，该计数装置能够与离心装置相匹配，因此能够将其整体置入离心装置中进行离心，使得待检测物沉底。然后利用自动计数仪进行计数分析。
在一些实施例中，载板200边缘可以设置有支撑边220，支撑边220向计数组件100的底部一侧延伸的高度大于计数组件的高度。支撑边220可以设置在载板200任意相对的两边，也可以设置在载板200的四边，放置载板200时，支撑边220底部可以直接与桌面、台面等 接触，以避免计数组件100的底部直接接触桌面、台面等而被磨花，影响计数分析。
在一些实施例中，计数装置(如主体、盖板和/或载板)的材料可以为透明度高且具有抗化学腐蚀能力的材料。例如，聚丙烯、聚苯乙烯等。
本申请实施例可能带来的有益效果包括但不限于：(1)计数装置可以为可清洗重复使用，也可以为一次性使用，用作一次性使用时操作简便，无需安装和清洗；(2)通过最少一次成像即可实现样品槽内样品全体积颗粒或细胞等的成像计数，检测快速高效，解决并消除了由于颗粒或细胞等的分布问题和/或采样有限问题而产生的计数误差；(3)单次检测样品量可达500μL，并且极大地拓展了样品浓度检测的下限范围，最低可达0个/ml，尤为适合低浓度样品的计数检测。需要说明的是，本申请可根据应用需求调整计数组件及计数装置的尺寸，使其既可应用于微颗粒计数，也可用于各类细胞的精确计数和荧光分析，以及医疗临床分析中血细胞计数、尿液有形成分分析等领域。需要说明的是，不同实施例可能产生的有益效果不同，在不同的实施例里，可能产生的有益效果可以是以上任意一种或几种的组合，也可以是其他任何可能获得的有益效果。
显然，对于本领域技术人员来说，上述详细披露仅仅作为示例，而并不构成对本说明书的限定。虽然此处并没有明确说明，本领域技术人员可能会对本说明书进行各种修改、改进和修正。该类修改、改进和修正在本说明书中被建议，所以该类修改、改进、修正仍属于本说明书示范实施例的精神和范围。
同时，本说明书使用了特定词语来描述本说明书的实施例。如“一个实施例”、“一实施例”、和/或“一些实施例”意指与本说明书至少一个实施例相关的某一特征、结构或特点。因此，应强调并注意的是，本说明书中在不同位置两次或多次提及的“一实施例”或“一个实施例”或“一个替代性实施例”并不一定是指同一实施例。此外，本说明书的一个或多个实施例中的某些特征、结构或特点可以进行适当的组合。
此外，应当注意的是，为了简化本说明书披露的表述，从而帮助对一个或多个发明实施例的理解，前文对本说明书实施例的描述中，有时会将多种特征归并至一个实施例、附图或对其的描述中。但是，这种披露方法并不意味着本说明书对象所需要的特征比权利要求中提及的特征多。实际上，实施例的特征要少于上述披露的单个实施例的全部特征。
一些实施例中使用了描述成分、属性数量的数字，应当理解的是，此类用于实施例描述的数字，在一些示例中使用了修饰词“大约”、“近似”或“大体上”来修饰。除非另外说明，“大约”、“近似”或“大体上”表明所述数字允许有±20％的变化。相应地，在一些实施例中，说明书和权利要求中使用的数值参数均为近似值，该近似值根据个别实施例所需特点可以发生改变。在一些实施例中，数值参数应考虑规定的有效数位并采用一般位数保留的 方法。尽管本说明书一些实施例中用于确认其范围广度的数值域和参数为近似值，在具体实施例中，此类数值的设定在可行范围内尽可能精确。
最后，应当理解的是，本说明书中所述实施例仅用以说明本说明书实施例的原则。其他的变形也可能属于本说明书的范围。因此，作为示例而非限制，本说明书实施例的替代配置可视为与本说明书的教导一致。相应地，本说明书的实施例不仅限于本说明书明确介绍和描述的实施例。

Claims (23)

  1. 一种计数组件，其特征在于，所述计数组件包括主体，
    所述主体上设有内凹的样品槽，所述样品槽的底端为平面，所述样品槽由底端至顶端的横截面面积逐渐增大。
  2. 根据权利要求1所述的计数组件，其特征在于，所述样品槽为倒圆台形、倒椭圆台形或倒棱台形。
  3. 根据权利要求1或2所述的计数组件，其特征在于，所述样品槽的内壁与水平面的夹角小于90°。
  4. 根据权利要求3所述的计数组件，其特征在于，所述样品槽的内壁与水平面的夹角为45-60°。
  5. 根据权利要求1～4任一项所述的计数组件，其特征在于，所述主体为薄壁结构。
  6. 根据权利要求5所述的计数组件，其特征在于，所述薄壁结构的壁厚均一。
  7. 根据权利要求5所述的计数组件，其特征在于，所述主体为下部敞开的薄壁结构。
  8. 根据权利要求5所述的计数组件，其特征在于，所述主体为空心的薄壁封闭结构，或者实心结构。
  9. 根据权利要求1～8任一项所述的计数组件，其特征在于，所述计数组件还包括盖板，所述主体的上表面与所述盖板的下表面相贴合，所述盖板上开设有加样孔和排气孔。
  10. 根据权利要求9所述的计数组件，其特征在于，所述加样孔和所述排气孔在水平面上的投影与所述样品槽的底面在水平面上的投影不重叠。
  11. 根据权利要求9所述的计数组件，其特征在于，所述盖板的下表面的面积大于所述主体的上表面的面积。
  12. 根据权利要求9所述的计数组件，其特征在于，所述排气孔和/或所述加样孔的形状为圆形、椭圆形、矩形、菱形或不规则形状。
  13. 根据权利要求9所述的计数组件，其特征在于，所述加样孔和/或所述排气孔的边缘朝上一侧设置有第一凸起。
  14. 根据权利要求9所述的计数组件，其特征在于，所述加样孔和/或所述排气孔的边缘朝下一侧设置有第二凸起，所述第二凸起外侧与所述样品槽的内壁接触。
  15. 根据权利要求1～14任一项所述的计数组件，其特征在于，所述样品槽的底面小于相机的成像靶面。
  16. 根据权利要求15所述的计数组件，其特征在于，所述样品槽的底面直径为1.5-2.0mm。
  17. 一种计数装置，其特征在于，包括权利要求1～16中任一项所述的计数组件。
  18. 根据权利要求17所述的计数装置，其特征在于，所述计数组件包括盖板，所述计数装置还包括载板；所述载板上开设有至少一个与所述计数组件匹配的孔洞，所述孔洞的直径在所述主体的外径与所述盖板的直径之间。
  19. 根据权利要求18所述的计数装置，其特征在于，所述载板上开设有24、48或96个孔洞，每个所述孔洞中固定安装有所述计数组件。
  20. 根据权利要求19所述的计数装置，其特征在于，所述载板与所述计数组件一体成型，或者可拆分连接。
  21. 根据权利要求19所述的计数装置，其特征在于，所述载板边缘设置有支撑边，所述支撑边向计数组件的底部一侧延伸的高度大于所述计数组件的高度。
  22. 一种计数组件和/或计数装置在微粒计数分析领域中的应用，其特征在于，所述计数组件为如权利要求1～16中任一项所述的计数组件；所述计数装置为如权利要求17～21任一项所述的计数装置。
  23. 根据权利要求22所述的计数组件和/或计数装置在微粒计数分析领域中的应用，其特征在于，所述微粒为细胞、细菌或真菌。
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JP2004020535A (ja) * 2002-06-20 2004-01-22 Olympus Corp 粒子凝集判定用容器
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