WO2009128294A1 - Test object receiver and test apparatus equipped with said test object receiver - Google Patents

Abstract

A measurement unit (25g2) and an induction unit (25g1) for the liquid of the test object are disposed in a holding part that is disposed in the test object flow path of a test object receiver. A test unit (25h) is also disposed on the downstream side of the measurement unit (25g2) in the test object flow path. A plurality of projections (27) having an elliptic columnar shape of a plan-view ellipse (the transverse cross section has an elliptical shape) are erected from the bottom surface in the measurement unit (25g2) and test unit (25h). Said projections (27) protrude such that the major axial direction of the ellipse thereof is oriented in the flow path direction of measurement unit (25g2). Consequently, during the step of washing the measurement unit (25g2) and test unit (25h), twin vortices will not be formed at the rear end in the flow path direction of the projections (27) so that said units can be thoroughly washed and the S/N value can be improved when sensing the test object.

Description

Inspection target receptacle and inspection device provided with the inspection target receptacle

The present invention relates to a microfluidic component that quantifies the volume of a desired liquid or that is separated after quantification, for example, a test subject receptor for performing chemical, medical, or biological chemical analysis and the test. The present invention relates to an inspection apparatus including a target receiver.

Research is being conducted on methods for conducting chemical reactions such as higher-speed and higher-efficiency analysis and synthesis in a system in which various functions such as pumps, valves, and reactors are miniaturized and integrated on a substrate (chip).

As such a system, for example, a pump-less pump that rotates a test object receiver and uses a centrifugal force generated by the rotation to move and react the liquid in the flow path formed in the test object receiver. A micro system inspection apparatus has been proposed (see Patent Document 1). Since this device uses centrifugal force, a pump for driving the liquid is unnecessary, and a simple system can be obtained. In addition, due to the characteristics of centrifugal force, the device can be used for liquids at the same distance from the center of rotation. Have the advantage that the same force can be applied.

As another system, an example in which a metering capillary array is arranged in a test subject receptacle is known (see Patent Document 2).
JP 2006-208183 A Japanese Patent No. 3537813

However, in the invention described in Patent Document 1, a plurality of cylindrical protrusions are arranged in the inspection portion provided in the flow path to perform inspection by a biological or chemical reaction. When cleaning this protrusion with cleaning liquid, if the flow rate of the cleaning liquid in the flow path becomes high, a twin vortex is generated behind the protrusion, resulting in a closed flow field. There was a problem that the flow of the cleaning liquid from the part was not hit and cleaning could not be performed sufficiently. In this case, the back side of the columnar protrusion cannot be cleaned, and the S / N value at the time of detection is low, and there is a problem that the inspection performance is deteriorated.

The present invention has been made in order to solve the above-described problems, and improves the cleaning performance of the protrusions erected in the flow path of the liquid inspection object, thereby improving the S / N value when detecting the inspection object. It is an object of the present invention to provide an inspection object receiver that can be used and an inspection device including the inspection object receptor.

According to the present disclosure, there is provided an inspection object receiver used for an application for inspecting a liquid inspection object, and a flow path that moves the inspection object through a predetermined path, and a plurality of protrusions at least in the flow path A protrusion forming portion disposed in part, wherein a diameter of the protrusion in a direction orthogonal to the extending direction of the flow path is shorter than a diameter of the protrusion in the extending direction of the flow path. A test recipient is provided.

Moreover, the cross section of the protrusion may be elliptical, and the major axis thereof may be parallel to the extending direction of the flow path.

Further, the cross section of the protrusion may have a spindle shape in which the downstream end in the extending direction of the flow path is narrower than the upstream end.

In addition, the inspection target receptacle of the present disclosure rotates about a predetermined center portion, and the extending direction of the flow path is the direction of centrifugal force when the inspection target receptacle is rotated. There may be.

Further, the projection forming unit may include at least a measuring unit that arranges a plurality of projections between the projections at an interval in which the inspection target spreads by capillary action and measures a predetermined amount of the inspection target. good.

Further, the protrusion forming part may be composed of at least an inspection part that arranges a plurality of protrusions in the flow path and inspects by a biological or chemical reaction.

Further, at least one reagent introduction part for introducing a reagent may be provided between the measurement part and the inspection part in the flow path.

Further, according to the present disclosure, the inspection object receiver, a rotating unit that rotates the inspection object receiver so that the inspection object flows along the flow path by centrifugal force, and an operation of the rotation unit are performed. An inspection apparatus including a control unit for controlling is provided.

3 is a side view illustrating a configuration of a rotating part of the inspection apparatus 1. FIG. It is a top view showing the structure of the main-body part 37 in the test object receiver 3. FIG. It is a top view showing the structure of the lid | cover 39 in the test object receiver 3. FIG. FIG. 3 is a cross-sectional view taken along line II in FIG. FIG. 3 is a cross-sectional view taken along the line II-II in FIG. 5 is an enlarged view of an inspection pattern configuration unit (flow path) 25. FIG. 4 is a partially enlarged view of an inspection pattern configuration unit 25. FIG. It is the elements on larger scale of the measurement part 25g2 and the guidance | induction part 25g1. It is an expansion perspective view of the part of protrusion 26 of guidance part 25g1. It is a longitudinal cross-sectional view of the to-be-inspected receptacle 3 in a state where a lid 39 is placed. It is a longitudinal cross-sectional view of the to-be-inspected receptacle 3 in a state where a lid 39 is placed. It is a schematic diagram which shows a three-dimensional cylindrical cross section. It is a schematic diagram which shows the flow which passes a three-dimensional cylinder row | line | column. It is the graph which showed the flow-velocity distribution between wall z = 0 and 2 in the place away from the column of cylinders. It is the graph which showed the flow-velocity distribution in the various positions inside a cylinder row. It is a schematic diagram which shows the shape of a calculation area | region. It is a stream diagram which showed the flow which hits a cylinder row | line | column in the case of high flow velocity. It is a graph which shows the velocity distribution of inflow and outflow. It is a contour map of the flow direction component of velocity. It is a contour map of a component perpendicular to the velocity flow. It is a graph which shows the flow-velocity distribution in the cross section with the cylinder of the 2nd row. It is a graph which shows the flow-velocity distribution in the cross section between the cylinder of the 1st row and the 2nd row. It is a streamline diagram of the flow which hits a column of cylinders when the flow velocity is fast. It is a graph of the velocity distribution in an inflow place. It is a graph of the velocity distribution in an outflow place. It is a figure of the experiment around one cylinder (dye which flowed out from the cylinder surface, Re = 28.4 (the fluid inside the dye does not come out of it)). It is a figure which shows the streamline of the cylinder row | line front of FIG. It is a contour map of the flow direction component of a flow velocity vector. It is a contour map of a component perpendicular to the flow of the flow velocity vector. It is the graph which showed flow velocity distribution in a section with a cylinder of the 2nd row. It is the graph which showed the flow-velocity distribution in the cross section with the cylinder of the 1st row and the 2nd row. FIG. 6 is a streamline diagram of calculation results when an elliptical column is used in place of a cylinder in order to eliminate a twin vortex region behind the cylinder during cleaning. FIG. 6 is a streamline diagram passing through the first column to the third column of an elliptic cylinder column. FIG. 11 is a plan view of a modification of the protrusion 27. It is the image which showed the detection result in a cylinder, an elliptical column, and a tear-drop column.

The best mode for carrying out the present invention will be described with reference to the drawings. First, the configuration of the inspection apparatus 1 in the present embodiment will be described with reference to FIG. As shown in FIG. 1, the inspection apparatus 1 includes a disk-shaped inspection object receptacle 3 made of polystyrene resin and a rotating unit 5. As shown in FIG. 1, the rotating unit 5 includes a chuck unit 15, a rotating shaft 17 attached to the center of the lower surface of the chuck unit 15, a rotating motor 19 that rotationally drives the rotating shaft 17, and a rotating shaft 17. And a cover portion 23 attached to the bearing 21. An opening (not shown) is formed on the upper surface of the chuck portion 15. The opening communicates with a suction pump (not shown), and when the suction pump suctions, the lower surface of the inspection target receptacle 3 and the upper surface 15b of the chuck portion 15 are in close contact with each other so as not to be relatively movable. That is, the chuck portion 15 and the inspection object receiver 3 rotate together.

When the rotation motor 19 is driven in a state where the inspection object receiver 3 is fixed to the chuck portion 15 so that the center of the inspection object receiver 3 is coaxial with the rotation shaft 17, the inspection object reception body 3 is moved to the rotation shaft 17 (that is, The test object is rotated about the center of rotation (receiver 3). The rotating unit 5 is connected to a control unit (not shown) having a CPU, a ROM, a RAM, and the like. The operation of the rotating unit 5 is controlled by a program stored in a ROM (storage medium) of the control unit.

Next, the configuration of the inspection object receptacle 3 will be described with reference to FIGS. The inspection object receptacle 3 includes a main body portion 37 shown in FIG. 2 and a lid 39 shown in FIG. The main body portion 37 is formed by forming eight sets of inspection pattern constituent portions (flow paths) 25 on one surface of a disk-shaped member. As shown in FIG. 4, these inspection pattern constituting portions 25 and the like are grooves having a substantially rectangular cross section with an upper opening. FIG. 4 is a cross-sectional view taken along the line II in FIG.

As shown in FIGS. 2 and 6, the inspection pattern constituting unit 25 branches from the downstream portion 25 c that is a groove having a constant width and extends in the radial direction of the main body portion 37, and the peripheral portion of the main body portion 37. An upstream portion 25b that is a groove having a constant width extending in a direction, an introduction portion 25a that is a saddle-shaped recess, and a downstream portion 25c that is connected to the upstream portion 25b and whose longitudinal direction extends in the radial direction of the main body portion 37 And connected to the downstream portion 25c on the outer peripheral side of the main body portion 37 rather than the position where the second introduction portion 25d which is a substantially tulip-shaped recess and the upstream portion 25b branches, connected on the inner peripheral side of the main body portion 37. The third introduction part (reagent introduction part) 25e, which is a substantially eggplant-shaped concave part, and the liquid reservoir part 25f, which is an arc-shaped concave part extending in the circumferential direction, is connected to the downstream part 25c at the outermost periphery.

Here, the upstream portion 25b extends in the circumferential direction of the main body portion 37 (lateral direction in FIGS. 2 and 6), and the downstream portion 25c extends in the radial direction of the main body portion 37 (vertical direction in FIGS. 2 and 6). Yes. Therefore, in the portion where the upstream portion 25b and the downstream portion 25c are connected, the angle formed by the upstream portion 25b and the downstream portion 25c is a right angle. Further, the liquid reservoir portion 25 f reaches the outermost periphery of the main body portion 37. Further, in the second introduction part 25d and the third introduction part 25e, the end on the side connected to the downstream part 25c is the outer peripheral side in the main body part 37, and the opposite end is the inner peripheral side in the main body part 37. It is provided as follows.

As shown in FIG. 6, the entire upstream portion 25b, the portion of the introducing portion 25a adjacent to the upstream portion 25b, and the portion of the downstream portion 25c adjacent to the upstream portion 25b (the hatched portion in FIG. 6). The holding part 25g is formed. As shown in FIG. 7, the holding part 25g is divided into a measuring part 25g2 in the downstream part 25c and a guiding part 25g1 which is the other part.

As shown in FIGS. 5, 7, and 8, the guide portion 25 g 1 of the holding portion 25 g is a portion in which a large number of protrusions 26 having a cylindrical shape with a diameter of 30 μm are erected from the bottom surface of the inspection pattern constituting portion 25. . Further, as shown in FIG. 8, the measuring unit 25 g 2 of the holding unit 25 g has a large number of protrusions 27 having an elliptical columnar shape (elliptical cross section) in plan view from the bottom surface of the inspection pattern constituting unit 25. Part. The protrusion 27 is an elliptical columnar protrusion whose major axis direction is directed to the flow path direction (vertical direction in FIG. 8) of the measuring portion 25g2. That is, the diameter of the protrusion 27 in the direction orthogonal to the extending direction of the inspection pattern constituting part (flow path) 25 is shorter than the diameter of the protrusion 27 in the extending direction of the flow path. Therefore, it is difficult for the liquid flowing through the measuring portion 25g2 to form a twin vortex on the downstream side of the projection 27 in the flow path direction.

Further, as shown in FIG. 8, the protrusions 26 and 27 in the holding portion 25g are regularly arranged in a staggered pattern, and the interval between the protrusions is an interval at which the liquid test object spreads by capillary action. 8 and 9, the axial direction of the individual protrusions 26 and 27 in the holding portion 25g is a direction perpendicular to the bottom surface of the upstream portion 25b, and the height of the protrusion is a groove in the upstream portion 25b. The same as the depth of. Therefore, the upper surface of the protrusion is on the same plane as the portion other than the inspection pattern constituting portion 25 in the main body portion 37. The holding portion 25g is subjected to a hydrophilic treatment by plasma treatment throughout.

As shown in FIG. 6, the holding portion 25 g protrudes into the introduction portion 25 a, and the introduction portion 25 a has a region on the inner peripheral side (upper side in FIG. 6) and an outer peripheral side (upper side in FIG. 6). It is divided into the lower area in FIG. However, these two regions communicate with each other through a gap (cutout portion) 25k between the holding portion 25g and the opposite side surface of the introduction portion 25a.

Further, as shown in FIGS. 6 and 8, the portion of the downstream portion 25c from the downstream side of the portion connected to the third introduction portion 25e to the liquid reservoir portion 25f (hatched in FIG. 6). The portion) is provided with an inspection unit 25h. The inspection unit 25h has a large number of protrusions 27 having an elliptical column shape having an ellipsoidal shape in plan view (an elliptical cross section) from the bottom surface of the inspection pattern constituting unit 25, like the measuring unit 25g2 of the holding unit 25g. Part. The protrusion 27 is a protrusion in which the major axis direction of the ellipse is directed in the flow path direction (vertical direction in FIG. 8) of the measuring portion 25g2. That is, the diameter of the protrusion 27 in the direction orthogonal to the extending direction of the inspection pattern constituting part (flow path) 25 is shorter than the diameter of the protrusion 27 in the extending direction of the flow path. Therefore, it is difficult for a twin vortex to be formed on the downstream side of the projection 27 in the flow path direction by the liquid flowing through the inspection portion 25h. However, the interval between the protrusions constituting the inspection portion 25h becomes narrower toward the outside of the main body portion 37 (not shown).

Between the inspection part 25h and the holding part 25g, as shown in FIGS. 6 to 8, an intermediate part 25n, which is a region where no protrusion is formed, is formed. Moreover, as shown in FIG. 6, in the introduction part 25a, the pillar part 25i is provided in the part of about 1/3 from the outer peripheral side. The pillar portion 25i is formed by arranging a large number of protrusions on the bottom surface of the inspection pattern constituting portion 25, similarly to the holding portion 25g. The pillar portion 25i divides the introduction portion 25a into two regions. That is, the pillar portion 25i is a portion communicating with the upstream portion 25b on the inner peripheral side from the pillar portion 25i, and is located on the outer peripheral side from the pillar portion 25i, and is separated from the upstream portion 25b by the pillar portion 25i. It is divided into 25a2 which is a part.

Further, as shown in FIG. 6, in the liquid reservoir portion 25f, a pillar portion 25j is provided so as to surround a portion connected to the downstream portion 25c in an arc shape with a certain distance. The pillar portion 25j is formed by arranging a large number of protrusions on the bottom surface of the inspection pattern constituting portion 25, similarly to the holding portion 25g. The pillar portion 25j divides the liquid reservoir portion 25f into two regions. That is, the pillar portion 25j is on the inner circumferential side from the pillar portion 25j and is a portion communicating with the downstream portion 25c, 25f1, and on the outer circumferential side from the pillar portion 25j, and is separated from the downstream portion 25c by the pillar portion 25j. 25f2, which is a part of the area.

Further, in the third introduction part 25e, a pillar part 25L is provided in the vicinity of the outlet connected to the downstream part 25c, and is separated from the downstream part 25c. The pillar portion 25L is formed by arranging a large number of protrusions on the bottom surface of the inspection pattern constituting portion 25, like the holding portion 25g.

As for the second introduction part 25d, as shown in FIG. 6, a pillar part 25m is provided on the downstream side, and is separated from the downstream part 25c. Similarly to the pillar portion 25L, the pillar portion 25m also has a large number of protrusions arranged on the bottom surface of the inspection pattern constituting portion 25.

Next, the lid 39 that constitutes the inspection object receptacle 3 together with the main body portion 37 will be described with reference to FIGS. 3, 10, and 11. As shown in FIG. 3, the lid 39 is a disk-shaped member having the same diameter as the main body portion 37, and is formed of a transparent synthetic resin plate. The lid 39 is attached to the surface of the main body portion 37 on which the inspection pattern constituting portion 25 is formed so that the centers thereof coincide with each other. By doing so, the upper part of the inspection pattern constituting part 25 of the main body part 37 is closed by the lid 39 as shown in FIGS. However, since the liquid reservoir 25f reaches the outermost periphery of the main body 37 as described above, even when the lid 39 is attached, the outer peripheral side of the liquid reservoirs 25f to 35f is outside the inspection object receptacle 3. Communicate. In the lid 39, a hole may be formed in a portion corresponding to the upper part of the liquid reservoir 25f.

Further, in the portion where the protrusion is provided, such as the holding portion 25g, the inspection portion 25h, and the pillar portions 25i and 25j, the upper surface of the protrusion comes into contact with the lid 39 as shown in FIG. Further, as shown in FIG. 3, the lid 39 is provided with a large number of minute holes 41. These holes 41 are positions (for example, the introduction part 25a, the second introduction part 25d, and the third introduction part) where it is necessary to introduce the inspection object and the reagent in the inspection pattern constituting part 25 when the lid 39 is attached to the main body part 37. 25e). As a result, the inspection object and the reagent can be supplied to the inspection pattern constituting unit 25 with the lid 39 attached to the main body portion 37.

1. Flow Analysis Next, the effect of changing the shape of the protrusion from the conventional cylindrical shape having a circular cross section (projection 26) to the elliptical column shape (projection 27) having an elliptical cross section as in the present disclosure. In order to confirm the confirmation, the result of the flow analysis by the computer will be described. A calculation grid calculation code for numerical calculation according to a prototype shape plan for carrying out this flow analysis was created, and several computer simulations were performed. Based on the simulation results, we conducted flow observation and flow field analysis in the flow path, compared flow fields under different conditions, examined prototype shapes, and based on that knowledge, The optimum design of the shape and form of the flow path) 25, the inspection section 25h, etc. was studied. We developed a numerical calculation program for observing the flow in the flow channel, especially numerical simulations of the flow field for different flow velocities on the protrusions (cylindrical rows) in the reaction region.

A fluid calculation under a complicated shape in which the flow velocity is relatively slow and a large number of columnar objects exist in the flow field, such as the receiving object 3 to be inspected, requires a lot of calculation time, and it is efficient to calculate the whole. Not right. For this reason, only the measurement part which has many cylinders in a flow path was taken out, and the following calculation was performed. As a calculation code, a standard calculation code for a two-dimensional or three-dimensional uncompressed unsteady Navier-Stokes equation for describing an incompressible fluid flow is used. The current calculation is to calculate the state after the rotation reaches a constant speed during both reaction and cleaning. Considering the shape and flow velocity, the theoretical flow dynamics can be obtained when the obtained flow field becomes steady. It can be expected from the discussion, and the calculation result is a steady flow field. The calculation code used is a non-stationary code corresponding to this system, which is because future development is taken into consideration. It is assumed that a constant pressure corresponding to the centrifugal force is applied at the inflow portion and the flow velocity portion, and the flow field imposes a unidirectional flow as a condition.

である。ここで、u は流速ベクトル、p は圧力、t は時間、ρは密度、μは粘性係数を表す。 また、 本計算の非圧縮流体シミュレーションにはよく用いられる、ＭＡＣ法（Ｍａｒｋｅｒ　Ａｎｄ　Ｃｅｌｌ法）を用いる。各計算量は、適当な長さ、時間、質量単位で規格化（無次元化）されているとする。このとき、慣性の大きさと粘性の大きさを表すレイノルズ数Re と呼ばれる無次元パラメータが定義される。ＭＡＣ法では、圧力を次式

で計算し、式（２．１）の流体の非圧縮性を保証している。ここで、ｕは速度、ｐは圧力、Δｔは時間刻み幅、添え字のｎは時間ステップである。また、Ｎａｖｉｅｒ－Ｓｔｏｋｅｓ方程式（２．２）の時間積分には、オイラーの陰解法を用いた。

ここで、空間微分の離散化には、式（２．４）の慣性項に３次精度の風上差分（Ｋａｗａｍｕｒａ－Ｋｕｗａｈａｒａ　スキーム）を用い、その他の項に２次の中心差分を用いた。ｘ方向の３次精度上流差分法は次のようになる。

ここで、ｆは物理量、ｉはｘ方向の格子点番号を示す。また、この式は、次式のようにまとめられる（α＝１）。

ここでα＝３である。
また、圧力のポアソン方程式の解法とＮａｖｉｅｒ－Ｓｔｏｋｅｓ方程式の陰解法には、擬似ＳＯＲ法を用いた。 2. Calculation code The governing equation of ordinary fluid is the incompressible Navier-Stokes equation

It is. Here, u is a flow velocity vector, p is pressure, t is time, ρ is density, and μ is a viscosity coefficient. The MAC method (Marker And Cell method), which is often used for the incompressible fluid simulation of this calculation, is used. It is assumed that each calculation amount is normalized (non-dimensionalized) by an appropriate length, time, and mass unit. At this time, a dimensionless parameter called Reynolds number Re representing the magnitude of inertia and the magnitude of viscosity is defined. In the MAC method, the pressure is

And the incompressibility of the fluid of formula (2.1) is guaranteed. Here, u is the speed, p is the pressure, Δt is the time step size, and the subscript n is the time step. Euler's implicit method was used for time integration of the Navier-Stokes equation (2.2).

Here, for discretization of the spatial differentiation, a third-order precision upwind difference (Kawamura-Kuwahara scheme) was used for the inertial term of Equation (2.4), and a second-order central difference was used for the other terms. The third-order precision upstream difference method in the x direction is as follows.

Here, f is a physical quantity, and i is a lattice point number in the x direction. Further, this equation can be summarized as the following equation (α = 1).

Here, α = 3.
The pseudo SOR method was used for solving the Poisson equation of pressure and implicitly solving the Navier-Stokes equation.

3. Calculation grid If a boundary-matching grid is used in this calculation, a grid must be generated each time the arrangement of the cylinders is changed, which is not efficient. The target of this study is the flow of low Reynolds number. In these flows, the change in the shape of the columnar object does not affect the calculation result of the drag so much. The cylinder is approximated by a polygon.

As an example, FIG. 12 shows a polygonal approximation of a cylindrical column of a 60-diameter grid. FIG. 12 is a schematic diagram showing the flow past the three-dimensional column. In FIG. 12, a circle to be approximated is indicated by an outer solid line, a circle approximated by a polygon actually used for calculation is indicated by a wavy line, and an inscribed circle of the polygon is indicated by an inner solid line. As shown in FIG. 12, in this calculation, the point inside the circle to be approximated is set inside the object. Then, the polygon used for the actual calculation in FIG. 12 is between the circles having a diameter of 60 and a diameter of 58. That is, if the diameter of the cylinder to be approximated is D, the approximated polygon is D and D × (number of grid points used to represent the diameter−2) / (number of grid points used to represent the diameter). Between.

These are compared with the analytical solution (Faxen 1946) of the drag coefficient of a two-dimensional flow that passes through a single cylinder placed between two parallel plates to determine the effective radius of the circle and investigate the degree of error. did. In this calculation, the processing method of the boundary condition of the velocity of the cylinder part is prepared as an array with 1 for the fluid part (outside the boundary) and 0 for the cylinder part including the boundary (inside the boundary). After calculating without distinguishing between the cylinder part and the cylinder part, the arrangement is multiplied and the velocity of the cylinder part is set to zero.

4). Calculation results and discussion 4.1 Investigation of the effect of the top and bottom walls To see the effect of the wall on the bottom surface of the cylinder, consider the four rows of three-dimensional cylinders in the center of the measurement section as shown in Fig. The calculation was performed assuming that FIG. 13 is a schematic diagram showing the flow past the three-dimensional cylindrical column. As a result, it can be said that the influence of the upper and lower walls is small as follows. The calculation area of FIG. 13 is divided into 900 × 66 × 20 calculation grids in the flow direction, the horizontal direction, and the vertical direction (x, y, z directions), and the calculation is performed using the difference method. When the rotation speed of the apparatus is constant, the flow in the flow path becomes steady at the current speed. In the calculation, a steady flow field is obtained by executing the calculation with a constant inflow velocity. In the horizontal direction, since many cylinders are arranged, periodicity is imposed. Also, using the knowledge of fluid mechanics that the fine shape of the object does not have a large effect on the flow in slow flow, the cylinder is expressed as a set of rectangular parallelepipeds with a square of 1/60 diameter in the horizontal plane. Yes. In the center plane of FIG. 13, a streamline (or a flow trajectory) of the fluid flowing on the plane is drawn. The color on each line indicates the flow velocity, and is faster when passing through the gap, and when passing through the area before and after the cylinder, the gap in the cross-sectional direction becomes larger, so the flow velocity is slower. The flow is from left to right. However, since the flow velocity is slow and the influence of the viscosity of the fluid is strong, the flow is almost symmetrical before and after the column, and it is difficult to guess the flow direction even when looking at the streamline in FIG. Yes. Looking at the flow between the cylinders, it can be seen that the flow is a flow field along the cylinders, and that the flow velocity is symmetric both before and after the cylinders.

Investigate the velocity distribution in the z direction to show the influence of the upper and lower walls of these flow fields. It is well known that when there is no cylinder, the flow is sandwiched between two flat plates, and unless the flow velocity becomes very high, a parabolic flow velocity distribution called a two-dimensional Poiseuille flow is obtained. 14 and 15 show flow velocity distributions along various vertical straight lines. FIG. 14 is a graph showing the flow velocity distribution between the walls z = 0, 2 at a location away from the cylinder row, and FIG. 15 is a graph showing the flow velocity distribution at various positions inside the column row. In each graph, the vertical axis (Z-axis) indicates the vertical distance along the axis of the cylinder, the lower wall is z = 0, and the upper wall is z = 2. The horizontal axis represents the flow velocity in the x direction. The flow velocity distribution of FIG. 14 that is sufficiently separated from the cylinder is a two-dimensional Poiseuille flow velocity distribution in which z = 0, 2 is a wall and the maximum flow velocity is obtained when z = 1. On the other hand, FIG. 15 shows flow velocity distributions along straight lines in the z direction at different positions of the x and y coordinates in the column. The average velocity along the straight line varies depending on the position, but unlike the outside of the column, the velocity increases rapidly from the flow velocity 0 at the wall z = 0, 2, so that 0.1 <z <1.9. It can be seen from the flow velocity distribution at any position that the velocity is almost constant in the range.

Note that FIG. 15 shows the flow velocity distribution between the walls z = 0, 2 on several straight lines in the column, but the marks on each line indicate that the x coordinate in the flow direction has the same value. The difference in the distribution of the lines of the marks indicates the difference in the horizontal direction, that is, the y coordinate. From this, it can be concluded that the horizontal arrangement is important for the distribution of the cylinders in the reaction region, and that changes in the vertical direction need not be considered unless the cylinders are tilted. The flow rate calculated here is slow. The Reynolds number, which is the ratio of the viscosity to the flow rate effect based on the diameter of the cylinder, is about Re = 0.01. However, from the analogy of other flow fields, it is expected that a substantially proportional result can be obtained even if the flow rate is increased and Re = 10. For this reason, the flow corresponding to the cylinder of this system is assumed to be a two-dimensional flow, and calculation is performed considering only the central part of the cylinder.

4.2 Evaluation of flow by change of flow velocity 4.1. Because the influence of the upper and lower walls is less than the three-dimensional simulation of, the majority of the z-direction around a large number of cylinders in a nearly uniform flow occupied by a two-dimensional flow with little change in the z-direction The state of the flow field was investigated by two-dimensional flow numerical analysis. At this time, the simulation was performed for two cases of flow rate (A) during reaction and (B) during washing. (B) is about 10 times the pressure difference with respect to (A). The flow field and the force applied to the cylinder were compared and examined. The calculation area is shown in FIG. FIG. 16 is a schematic diagram showing the shape of the calculation area. Consider a column with 11 rows in the flow direction and 18 rows in the direction across the flow path. Since the influence of the end of the column in the flow direction is considered to be not so strong, the number of column is less than the actual experimental device. In addition, since the flow path is narrow and the influence of the rotating Coriolis force is considered to be negligible, the flow is considered to be symmetric at the center of the flow path, so the area from the wall y = 0 to the central plane of the flow path is calculated. . However, the number of cylindrical columns in the flow direction is reduced to about 1/3 of the actual number in order to improve the calculation efficiency. Therefore, the centrifugal force due to the rotation for inducing the flow may be small, and a pressure difference due to the centrifugal force generated at about 1 / 2-1 / 3 of the actual number of rotations is given.

In the example shown in FIG. 16, the influence of the horizontal wall seems to be limited to the vicinity of the wall inside the column, but it affects the flow field of the entire flow path outside the column. This change is theoretically known in a slow flow (more strictly speaking, a low Reynolds number flow having a strong influence of viscosity) compared to the flow path width. For this reason, a calculation region of about 5 times the flow path width is provided before and after the column, and the outer flow is a Poiseuille flow. The calculation grid is 902 × 594, and 104 cylinders exist in the calculation area.

(A) In the case of slow flow rate at the time of reaction FIG. 17 shows the calculation results according to the flow rate at the time of reaction. FIG. 17 is a streamline diagram showing a flow corresponding to a column of cylinders when the flow velocity is high. The size of the cylinder and how it is arranged are matched to those of the current prototype system. When the pressure difference between the two ends is converted according to the shape of the test object receiver 3 (immuno disk), the corresponding rotation speed is about 400 rpm, which is considered to correspond to the actual rotation speed of about 1000 rpm. In FIG. 17, as shown in FIG. 13, streamlines from appropriate upstream positions are shown colored according to the magnitude of the flow velocity at each point. The flow flows from left to right, the lower surface is a wall, the upper surface is the center plane of the flow path, and the flow field of the entire actual flow path is twice as large as the figure folded back on the upper surface. For this reason, the flow near the upper surface is nearly green and the flow near the lower surface is blue, and the flow velocity is close to zero. As shown in FIG. 18, the flow that flows in and out is a Poiseuille flow, similar to the numerical simulation of I, and has a velocity that is proportional to the square of the distance from the wall that is the maximum velocity on the upper surface. . The Reynolds number is about Re = 4. FIG. 18 is a graph showing the velocity distribution of inflow and outflow. On the other hand, as shown in FIG. 17, it can be seen that near the cylinder row, the flow velocity is almost constant except near the wall. Comparing the streamlines flowing into and out of the cylinder row, the streamlines are not completely symmetrical, and the streamline slope is slightly changed, but basically the flow is almost the same.

Looking at the streamlines near the cylinders, they are almost the same except for the left, right, and lower cylinders in the column, and the streamlines flow along the cylinder so as to wrap the cylinder up and down. I understand that. For this reason, the cylinders are substantially symmetrical on the left, right, front and back sides. In a cross section without a cylinder, streamlines are slanted, but the flow velocity is almost constant. In addition, although the flow velocity is zero on the cylinder, the flow path becomes narrow in the cross section with the cylinder, so that the flow becomes faster, and the flow velocity is about twice as high as the flow speed in the cross section without the cylinder. In order to show the velocity field more clearly, a contour map of the velocity component in the flow direction is shown in FIG. 19, and a contour map of the velocity component in the direction perpendicular to the flow is shown in FIG. FIG. 19 is a contour map of the velocity direction component, and FIG. 20 is a contour diagram of the component perpendicular to the velocity flow.

In FIG. 19, the flow is only the flow component from the left to the right in the entire region, and contour lines with a high speed are shown in red and contour lines with a low speed are shown in blue. (The color bar starts with a negative value in order to color the contour line of velocity 0.) As with the streamline diagram, except for the cylinders in the boundary area, blue contour lines around each cylinder, It can be seen that the red contour lines are arranged in corresponding positions in the same shape. In FIG. 20, the upward component is represented by red contour lines, and the downward component is represented by blue contour lines. Since the flow is along the cylinder in the inner cylinder, if you take one cylinder, the red area is the top and the blue area is the bottom on the front of the cylinder, the blue area is the upper and the lower area is the red area behind the cylinder This pattern appears repeatedly, and the shape of the blue and red areas is almost the same.

In order to examine in detail the state of the flow in a cross section perpendicular to the flow, the flow velocity distribution of the cross section between the cross section passing through the center of the cylinder and the cylinder is shown in FIGS. FIG. 21 is a graph showing the flow velocity distribution in a cross section with the second row of cylinders, and FIG. 22 is a graph showing the flow velocity distribution in the cross section between the first row and the second row of cylinders. In each of FIGS. 21 and 22, the vertical axis represents the distance from the wall, and the horizontal axis represents the magnitude of the flow direction component of the velocity. FIG. 21 shows a flow velocity distribution in a cross section passing through the center of the second column from the inflow side. The flow velocity is 0 in the portion where the cylinder exists. In the second row, there is a semicircular cylinder on the wall. It can be seen that there is a nearly constant flow except for the cylinder near the wall. FIG. 22 shows the flow velocity distribution in the cross section between the first and second columns. At this time, the cross section has a non-zero flow velocity, but the maximum velocity component is about 1.6, and in FIG. 22, it is about ½ of the maximum velocity in FIG.

(B) In the case of a fast flow rate at the time of washing FIG. 23 shows the calculation results in accordance with the flow velocity at the time of washing. FIG. 23 is a streamline diagram of a flow hitting a column of cylinders when the flow velocity is high. In the example shown in FIG. 23, the size of the cylinder and the state of arrangement are the same as (A) according to the current prototype system. The pressure difference between the inflow and the outflow is 10 times, and the corresponding rotation speed is about 1200 rpm, which is considered to correspond to the rotation speed of about 3000 rpm in reality. The average flow velocity at the time of inflow is about 25 times that of (A). In FIG. 23, streamlines from appropriate upstream positions are shown colored by the magnitude of the flow velocity at each point. As in (A), the flow flows from left to right, the lower surface is a wall, the upper surface is the center plane of the flow path, and the flow field of the entire actual flow path is twice that of the figure folded at the upper surface. It is a size. For this reason, the flow near the upper surface is nearly green and the flow near the lower surface is blue, and the flow velocity is close to zero. As shown in FIGS. 24 and 25, the flow flowing in and out is a Poiseuille flow, but the flow does not return to the Poiseuille flow in the outflow portion as shown in FIG. This is because in FIG. 23, the left stream line entering the cylinder row is almost the same as (A), but the right stream line exiting from the circular stream row forms a wake region that is largely separated from the cylinder behind the cylinder. It is because it is doing. FIG. 24 is a graph of the velocity distribution at the inflow location, and FIG. 25 is a graph of the velocity distribution at the outflow location.

Referring to FIG. 23, when the streamline in the vicinity of the cylinders inside the cylinder row is seen, it can be seen that it is greatly different from the case of (A), except for the left end, the right end, and the lower side cylinder of the column row. It is almost the same shape. The streamline flows along the cylinder on the front surface of the cylinder, but is far away from the cylinder at the rear of the cylinder, and an area where the streamline does not enter is formed at the rear of the cylinder. Based on the diameter of one cylinder, the Reynolds number Re, which is a dimensionless parameter for the magnitude of the average flow velocity with respect to the viscosity of the fluid, is about 28. In the case of one cylinder placed in a uniform flow, in a region of 10 <Re <50 including this flow velocity, a pair of twin vortex regions are observed behind the cylinder, and Re = 28 as shown in FIG. Then, the size of the region is about the cylinder diameter. FIG. 26 is a diagram of an experiment around one cylinder (dye flowing out from the surface of the cylinder, Re = 28.4 (the fluid inside the dye surrounded by the dye does not go outside)). At the current speed due to the arrangement of the cylinder rows, the region of the twin vortex is suppressed and is approximately the radius of the cylinder. The flow state between the cylinders can be increased or decreased slightly. It seems that it does not change. FIG. 27 is a diagram showing streamlines in front of the column of columns in FIG. In the figure shown in FIG. 27, the vortex region is small because of the rear cylinder, but a twin vortex region can be seen. Further, the last streamline of the downstream cylinder is largely separated from the cylinder, and a twin vortex similar to that in FIG. 26 exists.

Since it is difficult to plot the flow lines inside the twin vortex, FIG. 28 shows a contour map of the flow direction component of the flow velocity vector, and FIG. 29 shows a contour map of the perpendicular component to the flow of the flow velocity vector. In FIG. 28, the flow component in the left-to-right direction is positive and indicated by red contour lines, the right-to-left flow component is negative, and values near zero and negative values are indicated by blue contour lines. Unlike FIG. 19 in (A), blue contour lines around each cylinder are slightly on the front and side surfaces of the cylinder, but occupy a large area behind the cylinder. In other words, it is thought that a flow toward the cylinder is created behind the cylinder, and twin vortices are formed. In addition, the red contour line has a shape that wraps around an isosceles triangle with the right side as the apex, and it can be seen that the flow wraps around the twin vortex behind the cylinder.

In the example shown in FIG. 29, the upward flow component is represented by red contour lines, and the downward flow component is represented by blue contour lines. Since the flow is along the cylinder in the inner cylinder, when one cylinder is taken, the red area is on the top and the blue area is on the bottom in front of the cylinder. However, as compared with FIG. 20, this pattern that spreads a lot between the cylinders is pressed against the front surface of the cylinder, and an area where no contour lines enter is formed behind the cylinder.

(A) and (B) Consideration of the flow field The streamlines inside the twin vortex formed behind the cylinder in the case of (B) as shown in FIGS. 26 to 29 are locally closed curves. Therefore, in the portion in contact with the blue region behind the cylinder in FIG. 26, the flow does not hit from the inflow portion. For example, in the case of cleaning the column of columns, the flow becomes faster when the rotational speed of the system is increased. Although cleaning of the front and side surfaces is performed efficiently, cleaning the back of the cylinder is expected to cause a state that cannot be performed in many cleaning times.

On the other hand, in the slow flow field like the state of (A), since the flow flows along the cylinder, it can be considered that the reagent or the like adheres to the entire cylinder and the reaction occurs in the entire cylinder. When the system is rotated from a stationary state during cleaning, an unsteady flow field generated before the rotation becomes constant, or the same state as (A) occurs when the rotation is slow. Can flow in. Although it is conceivable to change the rotation speed during cleaning, twin vortices are a phenomenon seen in a wide Reynolds number range (inflow speed range), and it is considered difficult to cope with such a change in rotation speed.

Next, in order to investigate in detail the state of the flow in the cross section perpendicular to the flow, the flow velocity distribution of the cross section between the cross section passing through the center of the cylinder and the cylinder is shown in FIGS. FIG. 30 is a graph showing the flow velocity distribution in a cross section with the second row of cylinders, and FIG. 31 is a graph showing the flow velocity distribution in a cross section with the first and second rows of cylinders. In both the examples shown in FIGS. 30 and 31, the vertical axis represents the distance from the wall, and the horizontal axis represents the magnitude of the velocity direction component. In the example shown in FIG. 30, the flow velocity distribution of the cross section which passes along the cylinder center of the 2nd row from an inflow side is shown. The flow velocity is 0 in the portion where the cylinder exists. In the second row, there is a semicircular cylinder on the wall. The flow has a substantially constant velocity except for a cylinder close to the wall, but the influence of the wall is stronger and the flow velocity is slightly faster away from the wall than in the case of (A). In addition, due to the effect of twin vortices, the velocity peak region between each cylinder is wide, and two peaks are visible. In the example shown in FIG. 31, the flow velocity distribution in the cross section between the first row and the second row of cylinders is shown. This place is outside the twin vortex, and all the velocities in the flow direction are positive. Compared with FIG. 21 in (A), the speed fluctuation remains large, and the maximum speed is about 80% of the value in FIG. 29, which is about 1/2 of the maximum speed on the side surface of the cylinder as in (A). It is not. It can also be seen that the flow velocity near the wall is particularly slow.

(C) In the case of an elliptical cylinder with a fast flow rate at the time of cleaning Next, a case where an elliptical cylinder array was used like the protrusion 27 of the present embodiment was simulated. FIG. 32 is a streamline diagram of calculation results when an elliptical column is used instead of a cylinder in order to eliminate the twin vortex region behind the cylinder during cleaning. When the elliptic cylinder row shown in FIG. 32 is used, the flow calculation conditions are the same as in (B), the center of the cylinder corresponding to the center of the elliptic cylinder is located, and the minor axis is the cylinder of (B). The diameter and the major axis are 1.7 times the diameter of (B). The flow is asymmetrical in the longitudinal direction, but the wake twin vortex region is small except for the last column of cylinders.

Next, FIG. 33 shows a streamline diagram passing through the first column to the third column of the elliptical column. In the example shown in FIG. 33, it can be seen that, unlike the example shown in FIG. 27, the flow flows along the cylinder and does not make a twin vortex behind the elliptical column. However, it was observed that a twin vortex was formed in the last cylinder of the channel. However, in an actual disk, the elliptical column is about three times the calculation, and the flow is always in one direction due to centrifugal force. Therefore, the influence of the last elliptical column does not contribute to the reaction of other cylinders. It is not considered.

As described above, in the analysis by the computer simulation, the flow field analysis was performed between the projections provided in the flow path of the inspection target receptacle 3 having a conventional cylindrical shape and the elliptical column shape. At this time, the flow field was analyzed in consideration of the rotational motion of the prototype 3 to be inspected. The three-dimensional calculation shows that the flow deviation in the axial direction of the protrusion is small. Also, in the calculation of the flow field when the horizontal section is taken out, the rotational speed is different between the reaction time and the washing time, so there is a greater difference in flow velocity than the centrifugal force difference, which greatly changes the flow around the protrusion. It was shown that. With a conventional cylindrical protrusion, the flow flows without stagnation at the time of reaction, but at a high rotational speed during cleaning, a vortex region is formed behind the pillar, and the flow stagnates. In actual operation, since the rotation speed is continuously increased, the flow may enter the rear of the cylinder, but it is considered that the rear of the protrusion is difficult to clean. For this reason, calculation was performed when the protrusion was an elliptic cylinder like the protrusion 27 of the present disclosure, and it was shown that the stagnation of the flow can be suppressed except for the rearmost protrusion. For the correct evaluation of the last flow, it is necessary to consider the shape of the test object receiver 3, but the flow is slow because the flow is unidirectional and the fluid area behind the last pillar is widened. Therefore, it is not considered important.

Next, as an example of a method of using the test target receptor 3 having the above-described configuration, a method for accurately determining the degree of anemia by quantifying the transferrin concentration in human blood by ELISA will be described.

(I) Immobilization of antibody to test target receptor 3 For each test pattern constituting unit 25, a test solution 25h is diluted with sodium carbonate buffer solution of transferrin antibody derived from Goat (0.05M NaHCO3, pH 9.6, 10 μg). / Ml, hereinafter referred to as a primary antibody solution).

Specifically, the test object receiver 3 is attached to the test apparatus 1 as shown in FIG. 1, and 15 μL of the primary antibody solution is injected into each of the third introduction parts 25e of each test pattern constituting part 25. The injection into the third introduction part 25e is performed through a hole 41 (see FIG. 3) formed in the lid 39. The injected solution remains inside the third introduction part 25e by the pillar part 25L.

Thereafter, the test object receiver 3 is rotated counterclockwise when viewed from the direction of FIG. 2 at a rotational speed of 100 to 3000 rpm (rotational speed R2). Then, by the centrifugal force, the primary antibody solution flows out from the third introduction part 25e, enters the downstream part 25c, flows through the inspection part 25h, and reaches the liquid reservoir part 25f. Since the outer peripheral side of the liquid reservoir 25f communicates with the outside as described above, the primary antibody solution is discharged to the outside. Thereafter, the rotation of the inspection object receptacle 3 is stopped.

(Ii) Blocking For each inspection pattern constituting unit 25, 15 μL of a blocking solution (50 mM Tris, 0.14M NaCl 1% BSA, pH 8.0) is allowed to flow through the inspection unit 25h.

Specifically, first, the primary antibody solution flowed in the step (i) is removed from the inspection unit 25h by rotating the test target receptor 3 at a rotation speed of 150 to 15000 rpm (rotation speed R1). Next, 15 μL of the blocking solution is injected into each of the third introduction parts 25e, and the test object receiver 3 is rotated counterclockwise at a rotation speed of 100 to 3000 rpm (rotation speed R2). Then, like the primary antibody solution in the step (i), the blocking liquid enters the downstream part 25c from the third introduction part 25e, flows through the inspection part 25h, and is discharged to the outside from the liquid reservoir part 25f. .

</ RTI> Through the steps so far, the primary antibody is fixed to the inspection unit 25h. Thereafter, the following cleaning process is performed. In the cleaning process, first, each test pattern constituting unit 25 and a cleaning liquid (50 mM Tris, 0.14 M NaCl, 0.05% Tween 20, pH 8.0, hereinafter referred to as a cleaning liquid) are injected. The place where the cleaning liquid is injected is the third introduction part 25e, and the injection is performed through the corresponding holes 41 (FIG. 3).

Next, the inspection object receiver 3 is rotated at a rotational speed of 100 to 3000 rpm (rotation speed R2) to fill the inspection unit 25h with the cleaning liquid, and then the inspection object receiver 3 is rotated to 150 to 15000 rpm (rotation speed R1). To remove the cleaning liquid from the inspection section 25h. The removed cleaning liquid is discharged to the outside through the liquid reservoir 25f.

Note that the protrusions 27 erected on the measuring unit 25g2 and the inspection unit 25h are elliptical columns erected so that the major axis direction thereof is parallel to the flow direction of the cleaning liquid. Therefore, even in the cleaning step, twin vortices are not generated on the rear side of the projection 27 in the flow path direction, and the rear side of the projection 27 can be sufficiently cleaned. Therefore, the S / N value at the time of inspection can be kept good.

(Iii) Capture by antigen-antibody reaction to be examined Here, the test object is transferrin. 1 μL each of a solution (hereinafter referred to as an antigen solution) prepared by transferring Tris buffered saline (50 mM Tris, 0.14 M NaCl, 1% BSA, 0.05 Tween 20, pH 8.0) to a concentration of 125 ng / ml Flow to inspection pattern configuration unit 25.

Specifically, for each of the test pattern constituting units 25, 1 μL of the antigen solution is injected into the introducing unit 25a. At this time, the antigen solution is injected through a hole 41 (see FIG. 3) provided in the lid 39 so as to correspond to the introduction portion 25a. The hole 41 is provided in the introduction portion 25a on the inner peripheral side with respect to the holding portion 25g that has entered the introduction portion 25a. Therefore, the antigen solution is supplied to the inner peripheral side of the introduction part 25a with respect to the holding part 25g.

Supplied antigen solution spreads inside the holding part 25g by capillary action. At this time, the amount of the antigen solution that spreads in the holding part 25g is equal to the volume of the gaps between the many protrusions constituting the holding part 25g. Next, when the test object receiver 3 is rotated at a rotational speed of 100 to 3000 rpm (rotation speed R2), the antigen solution held in the holding part 25g enters the downstream part 25c and advances through the inspection part 25h. To the outer edge. Thereafter, the rotation of the inspection object receptacle 3 is stopped.

At this time, in the inspection pattern constituting unit 25, only the one held in the weighing unit 25g2 out of the holding unit 25g flows to the inspection unit 25h, and the one held in the guiding unit 25g1 remains as it is. This is because the upstream portion 25b where the guide portion 25g1 is located is along the circumferential direction of the main body portion 37, so that the force to flow the inspection target along the inspection pattern constituting portion even if the inspection target receiver 3 is rotated. This is because does not work. The same applies to the inspection pattern configuration unit 25. In addition, among the antigen solutions injected into the introduction part 25a, those that are not held in the holding part 25g but remain in the introduction part 25a are included in the holding part of the introduction part 25a when the test object receiver 3 is rotated. Since it is carried to the outer peripheral side rather than 25g and is captured by the pillar part 25i, it does not flow into the inspection part 25h. Therefore, the antigen solution flowing into the test unit 25h is only a fixed amount held in the holding unit 25g.

Through this process, the antibody immobilized on the test section 25h captures transferrin. Thereafter, the inspection section 25h is cleaned by the following process using the same cleaning liquid as in (ii) above. In the inspection pattern constituting unit 25, the place where the cleaning liquid is injected is the second introduction unit 25d. The cleaning liquid is injected through the corresponding holes 41 (see FIG. 3).

(Iv) Binding of labeled antibody For each of the test pattern constituting units 25, the user puts the HRP-labeled Goat-derived transferrin antibody in Tris-buffered saline (50 mM Tris, 0.14 M NaCl). 10 μL each of a solution (hereinafter, secondary antibody solution) prepared to a concentration of 10 ng / ml with 1% BSA, 0.05 Tween 20, pH 8.0).

Specifically, the user injects 10 μL of the secondary antibody solution into each of the third introduction parts 25e of each test pattern constituting part 25, so that the test object receptor 3 is rotated at a rotation speed (rotation of 100 to 3000 rpm). Rotate counterclockwise by the number R2). Then, due to the centrifugal force, the secondary antibody solution enters the downstream portion 25c from the third introduction portion 25e, flows through the inspection portion 25h, and reaches the outer peripheral end.

In this step, Goat-derived transferrin antibody binds to the transferrin captured in (iii) above. Thereafter, the inspection section 25h is cleaned by the same cleaning process as in (iii).

(V) Quantification of inspection target The user sends ABTS phosphate-citrate solution (0.05 M sodium phosphate, 0.05 M) as a substrate solution to each inspection pattern constituent unit 25 for each inspection pattern 25 h. Citric acid) and hydrogen peroxide solution (hereinafter referred to as a color developing solution) are allowed to flow to develop the transferrin to be tested.

Specifically, the user injects the coloring solution into each of the third introduction parts 25e of each inspection pattern constituting part 25, and the inspection object receiver 3 is rotated at a rotational speed of 100 to 3000 rpm (rotational speed R2). Rotate counterclockwise. Then, due to centrifugal force, the coloring solution enters the downstream portion 25c from the third introduction portion 25e, flows through the inspection portion 25h, and reaches the outer peripheral end. Thereafter, the test object receiver 3 is applied to a fluorescence analyzer, the image is captured by a scanner, and the computer on which the darkness analysis software is activated digitizes the degree of color development.

Next, the effect produced by the test subject receptacle of the present embodiment will be described. In the inspection object receiver 3 of the present embodiment, when a liquid inspection object introduced into the inspection pattern constituting unit 25 is brought into contact with the holding unit 25g, a predetermined amount of the inspection object is absorbed into the holding unit 25g by capillary action. The Thereafter, if the centrifugal force generated by rotating the test object receptacle 3 is applied to the test object absorbed by the holding part 25g, the test object absorbed by the holding part 25g can be taken out.

At this time, the amount of the test object once absorbed by the holding portion 25g is a certain amount because it is the volume of the entire holding portion 25g holding portion minus the volume of the protruding portion (that is, the volume of the gap between the protrusions). It becomes. Therefore, if the above operation is performed using the inspection object receiver 3 of the present embodiment, a predetermined amount of inspection object can be measured.

Further, according to the inspection object receptacle 3 of the present embodiment, a minute amount of the order of nL to μL can be accurately measured. Furthermore, since the test object receiver 3 of the present embodiment can be manufactured in one step by using a resin injection molding method, it is not necessary to separately manufacture a microvalve and perform high-precision assembly. Therefore, the inspection object receiver 3 is easy to manufacture, has high productivity, and has a low manufacturing cost.

The inspection object receptacle 3 according to the present embodiment can inspect the inspection object measured as described above by a biological or chemical reaction in the inspection unit 25 h provided in a part of the inspection pattern configuration unit 25. .

Further, in the test object receptacle 3 of the present embodiment, the protrusions 27 that are erected on the measuring part 25g2 and the inspection part 25h are oval erected so that the major axis direction thereof is parallel to the flow direction of the cleaning liquid. It is a pillar. Accordingly, even in the cleaning process, twin vortices are not generated on the rear side in the flow path direction of the protrusion 27, and the rear side of the protrusion 27 can be sufficiently cleaned. Therefore, the S / N value at the time of inspection can be kept good.

In addition, it is good also considering the wall surface of the area | region corresponding to the pillar part 25i in the introduction part 25a as hydrophobic instead of providing the pillar part 25i. Since the hydrophobic region makes it difficult to pass through the inspection object, the same effect as described above can be obtained. In order to make it hydrophobic, for example, a treatment such as coating with a fluorine-containing compound may be performed. Alternatively, the wall surface may be formed of a material having a hydrophobic surface (for example, a fluororesin).

Thereby, the inspection object can be introduced into the inner peripheral side of the introduction part 25a with respect to the holding part 25g, and the inspection object can be absorbed by the holding part 25g. Then, the inspection object remaining without being absorbed by the holding portion 25g can be moved to the outer peripheral region of the introduction portion through the notch portion. If a centrifugal force is applied to the inspection object that has entered the outer peripheral area so as to move away from the holding part 25g, an excessive inspection object will not flow into the holding part 25g.

The inspection object receptacle 3 of the present embodiment has an inspection part 25h in which a plurality of protrusions 27 are arranged in each inspection pattern constituent part 25. As a result, the inspection target obtained by measuring a predetermined amount using the holding unit 25g can be moved to the inspection unit 25h, and an inspection based on a biological or chemical reaction can be performed. Since the inspection unit 25h includes a plurality of protrusions 27, the surface area is large and the detection sensitivity can be increased. Further, since the protrusion 27 is an elliptical column protrusion standing in a direction parallel to the flow path direction of the inspection portion 25h (flow direction of the inspection object), the flow path of the protrusion 27 is used in the cleaning process. The protrusion 27 can be sufficiently cleaned without generating a twin vortex on the rear side in the direction (flow direction to be inspected). Therefore, the S / N ratio of detection sensitivity can be increased.

Further, when a centrifugal force is applied to the inspection object held in the weighing unit 25g2 that is a part of the holding unit 25g so as to go to the inspection unit 25h, the object is held in the guide unit 25g1 that is a part of the holding unit 25g. A centrifugal force in a direction away from the inspection unit 25h is applied to the inspection target.

In this way, only the inspection object held in the weighing unit 25g2, which is a part of the holding unit 25g, is moved to the inspection unit 25h with good reproducibility and is held in the guide unit 25g1 which is a part of the holding unit 25g. It is possible to prevent the inspected object from flowing into the inspection unit 25h. That is, only the inspection object held in the weighing unit 25g2 which is a part of the holding unit 25g can be moved to the inspection unit 25h.

In this case, since it is possible to prevent the inspection object held in the guiding part 25g1 from flowing to the inspection part 25h, even if the upstream part 25b also serves as the introduction part 25a, the inspection part is unintentionally transferred from the introduction part 25a to the inspection part 25h. There is no such thing as flowing. Therefore, it is possible to save space by making the upstream portion 25b also serve as the introduction portion 25a.

In addition, since the inspection object receptacle 3 of the present embodiment has an intermediate part that does not have a protrusion between the holding part 25g and the inspection part 25h, the inspection object held by the holding part 25g is used. It does not flow to the inspection unit 25h against the intention of the person. If the inspection object receiver 3 is rotated and a sufficient centrifugal force is applied, the inspection object held by the holding part 25g can be passed through the intermediate part 25n to the inspection part 25h. Moreover, since the holding | maintenance part 25g is hydrophilically processed, it can absorb a test object smoothly.

In the test target receptacle 3 of the present embodiment, the pillar part 25L is provided in the vicinity of the outlet connected to the downstream part 25c in the third introduction part 25e, so that the reagent supplied to the third introduction part 25e is supplied. There is no such thing as flowing into the downstream portion 25c against the user's intention. Moreover, since the pillar part 25m is provided in the 2nd introduction part 25d near the exit connected with the downstream part 25c, the reagent supplied to the 2nd introduction part 25d is a downstream part against a user's intention. There is no such thing as flowing into 25c.

It should be noted that the present invention is not limited to the embodiment described above, and it goes without saying that the present invention can be implemented in various modes without departing from the present invention. For example, as shown in FIG. 34, the cross section of the protrusion 27 may be formed in a spindle shape in which the downstream end in the extending direction of the flow path is narrower than the upstream end.

Further, in the inspection pattern constituting part 25, a foreign substance removing part formed by arranging a plurality of different protrusions may be formed between the introducing part 25a and the holding part 25g. By doing so, even if the inspection object introduced into the introducing portion 25a contains foreign matter, the foreign matter can be removed by the foreign matter removing portion, so that the foreign matter does not enter the holding portion 25g.

Further, the inspection object receiver 3 can be used as a sorting device. That is, when the liquid object supplied to the introduction part 25a is brought into contact with the holding part 25g, a predetermined amount of the object is absorbed into the holding part 25g by capillary action. Thereafter, if a centrifugal force is applied to the object absorbed by the holding part 25g, the object absorbed by the holding part 25g can be taken out.

At this time, the amount of the object once absorbed by the holding portion 25g and then taken out is a constant amount because the volume of the protruding portion is subtracted from the entire volume of the holding portion 25g. Therefore, a predetermined amount of an object can be measured by performing the above operation using this sorting apparatus. Further, by using this sorter, a minute amount of the order of nL to μL can be accurately measured.

When used as a sorting device, the inspection unit 25h, the second introduction unit 25d, the third introduction unit 25e, and the like may be omitted. Further, the protrusion 26 is not limited to a column, and may be a cylinder. The protrusion 27 is not limited to an elliptic cylinder, and may be an elliptic cylinder.

An example of the effect of the inspection target receiver of this embodiment will be described below. FIG. 35 shows an image of transferrin detected using the same clinical test sample (human plasma) by forming pillars of elliptical cylinders, elliptical pillars, and teardrop-shaped pillars in each receptor for each of eight flow paths. Show. In this case, the fluorescent reagent AmplexRed (trademark) was used in place of the coloring reagent ABTS described in “(v) Quantification of test target”. Analysis software that digitizes the degree of fluorescence was activated on a computer, and the reproducibility CV value of each measurement data of the cylinder, the elliptical column, and the teardrop column was calculated from the image shown in FIG. CV (Coefficient of variation coefficient of variation) is the percentage of the average value of the standard deviation (SD), and is expressed as (standard deviation / average value) x 100 (%). As a result, the CV values were teardrop column: 5.4%, elliptical column: 5.3%, cylinder: 9.1%, and the pillar shape was measured by the elliptical column and teardrop column rather than the column. The effect of the test subject receiver of the present disclosure was recognized.

As described above, according to the test subject receptacle of the present embodiment, the diameter of the protrusion in the direction orthogonal to the extending direction of the flow path is shorter than the diameter of the protrusion in the extending direction of the flow path. Therefore, a twin vortex does not occur behind the protrusion, and the flow of the cleaning liquid from the inflow portion hits the back side of the cylinder with respect to the flow, so that sufficient cleaning can be performed.

In addition, since the cross section of the protrusion is elliptical and the major axis is parallel to the extending direction of the flow path, no twin vortex is generated behind the protrusion, and the flow is cylindrical. On the back side, the flow of the cleaning liquid from the inflow portion hits, and the cleaning can be performed sufficiently.

In addition, the cross section of the protrusion has a spindle shape in which the end on the downstream side in the extending direction of the flow path is narrower than the end on the upstream side, so twin vortices are generated behind the protrusion. However, the flow of the cleaning liquid from the inflow portion hits the back side of the cylinder with respect to the flow, and the cleaning can be sufficiently performed.

Further, the inspection object receiver rotates about a predetermined central portion, and the extending direction of the flow path is a direction of centrifugal force when the inspection object reception body is rotated. The twin vortex does not occur behind the protrusion even when the rotational speed of the test object receiver increases and the centrifugal force increases, and the flow of the cleaning liquid from the inflow part hits the back side of the cylinder against the flow, Can be cleaned sufficiently.

In addition, the protrusion forming unit includes at least a plurality of protrusions arranged at intervals between the protrusions so that the inspection object spreads by capillary action, and measures a predetermined amount of the inspection object. A twin vortex does not occur behind the protruding portion erected on the measuring portion, and the flow of the cleaning liquid from the inflow portion hits the back side of the cylinder with respect to the flow, so that sufficient cleaning can be performed.

In addition, the protrusion forming part is composed of at least a test part that arranges a plurality of protrusions in the flow path and performs an inspection based on a biological or chemical reaction. The twins are arranged behind the protrusions standing on the measuring part. No vortex is generated, and the flow of the cleaning liquid from the inflow portion hits the back side of the cylinder with respect to the flow, so that the cleaning can be performed sufficiently.

Moreover, since at least one reagent introduction part for introducing a reagent is provided between the measuring part and the inspection part in the flow path, the reagent can be introduced from the reagent introduction part to the inspection part. .

Moreover, the above-described effects can be achieved in the inspection apparatus using the above-described inspection object receiver.

Claims (8)

A test object receiver used for inspecting a liquid test object,
A flow path for moving the inspection object through a fixed path;
A plurality of protrusions, and a protrusion forming portion arranged in at least a part of the flow path,
The inspection object receptacle, wherein a diameter of the protrusion in a direction orthogonal to the extending direction of the flow path is shorter than a diameter of the protrusion in the extending direction of the flow path. 2. The test object receptacle according to claim 1, wherein a cross section of the protrusion is elliptical, and a major axis thereof is parallel to an extending direction of the flow path. The inspection object according to claim 1, wherein a cross section of the protrusion has a spindle shape in which an end on the downstream side in the extending direction of the flow path is narrower than an end on the upstream side. Recipient. The inspection object receiver rotates about a predetermined center part, and
The test object receptacle according to any one of claims 1 to 3, wherein the extending direction of the flow path is a direction of centrifugal force when the test subject receptacle is rotated. The protrusion forming unit includes at least a plurality of protrusions arranged at intervals between the protrusions so that the inspection object spreads by capillary action and measures a predetermined amount of the inspection object. 5. The inspection target receptacle according to claim 1. The inspection according to any one of claims 1 to 5, wherein the protrusion forming portion includes at least an inspection portion that performs inspection based on a biological or chemical reaction by arranging a plurality of protrusions in the flow path. Target recipient. The test object receptacle according to claim 6, wherein at least one reagent introduction part for introducing a reagent is provided between the measurement part and the inspection part in the flow path. The test object receiver according to any one of claims 1 to 7,
A rotating unit that rotates the test object receptacle so that the test object flows along the flow path by centrifugal force;
An inspection apparatus comprising: a control unit that controls the operation of the rotating unit.

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