JP5268382B2 - Analytical device, analytical apparatus and analytical method using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analyzing device for performing measurement using a very small amount of a sample liquid and having a stirring mechanism for being easily made small in size. <P>SOLUTION: Since a measuring cell (40a) is formed so as to be extended in a centrifugal force acting direction, the analyzing device is miniaturized. Further, since a capillary tube area (47a) is formed to the sidewall positioned in the rotary direction of the measuring cell (40a) so as to be extended in the inner peripheral direction from the outer peripheral direction, a sufficient stirring effect is obtained by accelerating rotation to return the sample liquid of the capillary tube area (47a) to the measuring cell (40a) after the sample liquid of the measuring cell (40a) is sucked to the capillary tube area (47a) by reducing or stopping rotation and measurement can be performed using a very small amount of the sample liquid. <P>COPYRIGHT: (C)2009,JPO&amp;INPIT

Description

  The present invention relates to an analytical device used for analyzing a liquid collected from a living organism, an analytical apparatus and an analytical method using the analytical device, and more specifically, a sample liquid in a measurement cell in the analytical device, The present invention relates to a reagent stirring technique.

  Conventionally, as a method for analyzing a liquid collected from a living organism or the like, a method for analyzing using a device for analysis in which a liquid channel is formed is known. The analytical device can control the fluid using a rotating device, and utilizes centrifugal force to dilute the sample liquid, measure the solution, separate the solid component, transfer and distribute the separated fluid, Since a solution and a reagent can be mixed, various biochemical analyzes can be performed.

  As shown in FIGS. 24 (a) and 24 (b), the analytical device described in Patent Document 1 that transfers a solution using centrifugal force allows a sample solution to flow into the inflow path 114 from an inlet 116 using an insertion instrument such as a pipette. The sample liquid is injected and transferred to the measurement cell 115 by the rotation of the analysis device, the sample liquid is sucked up by the capillary force acting on the flow path 117 by the rotation reduction or stop, and the rotation is accelerated again to measure the sample liquid. The sample liquid and the reagent can be stirred by returning to the cell 115.

  As an analysis method of the sample solution, for example, there is a method of analyzing the reaction solution obtained by reacting the sample solution and the reagent by an optical technique. Some have a plurality of flow paths so that a plurality of items can be analyzed using one type of sample solution, or the same item can be analyzed for a plurality of types of sample solution. Specifically, FIG. 25 shows an analysis device found in Patent Document 2.

  This is because the sample liquid 123 injected into the liquid receiver 118 of the analysis device is transferred to the reaction chamber 119B of the analysis container via the flow path 119 of the analysis container by centrifugal force and capillary action, and the reaction chamber 119B. , The reagent 122 and the sample solution 123 set in the reaction chamber 119B are reacted to optically access the mixed solution in the reaction chamber 119B to read the color reaction of the mixed solution.

This analytical device is bonded to a base 120 (see FIG. 26A) having various recesses forming the flow path 119, the reaction chamber 119B and the like on the upper surface, and an adhesive layer on the upper surface of the base 120. The reaction chamber 119B is loaded with the reagent 122 by dropping a necessary amount of a liquid reagent into the reaction chamber 119B before the cover 121 is bonded to the upper surface of the base 120, and allowing it to dry naturally. Alternatively, after lyophilization, the base 120 and the cover 121 are bonded together with an adhesive layer.
JP 2006-145451 A JP 2004-150804 A

  However, in Patent Document 1, since the measurement cell 115 is arranged at a right angle to the centrifugal direction, when the sample liquid in the measurement cell 115 is optically measured, there is no sample liquid for filling the measurement cell 115. A large amount is required and it is difficult to reduce the amount of the sample solution.

  Further, since the configuration of the stirring mechanism including the inflow path 114 for stirring the sample solution and the reagent, the measurement cell 115, and the flow path 117 is U-shaped, it is formed between the inflow path 114 and the flow path 117. Area is formed as a useless space, and there is a problem that it is not suitable for downsizing of the analysis device.

  Further, in Patent Document 2, the reagent 122 is carried in the recess of the reaction chamber 119B. In this configuration, as shown in FIG. 26A, the reagent 122 dropped into the reaction chamber 119B is immediately after that or dried. During the process, as shown in FIG. 26 (b), the reagent 122 may adhere to the wall surface of the concave portion of the reaction chamber 119B, and may be dried in a state where the reagent concentration on the wall surface of the concave portion of the reaction chamber 119B is high.

  When the sample solution 123 is transferred to the reaction chamber 119B in this state, the reagent biased to the wall surface of the recess of the reaction chamber 119B becomes very difficult to dissolve, and there is a possibility that a uniform color reaction does not occur. This can be a major factor in the variation of measurement results. By making the area of the reaction chamber 119B sufficiently wider than the dropping amount of the reagent 122, contact with the concave portion of the reaction chamber 119B can be suppressed. Increase in liquid volume occurs.

  Further, in this flow path configuration, the sample solution 123 completely fills the reaction chamber 119B, so that the fluidity of the sample solution 123 is low, and it takes time to dissolve the reagent. Furthermore, the dissolved reagent However, it takes a long time to spread uniformly over one surface of the reaction chamber 119B, and there is a problem that the measurement time is enlarged and the variation in the measurement result becomes large.

  The present invention solves the conventional problems, and provides an analysis device having an agitation mechanism that can be measured with a small amount of sample liquid and can be easily miniaturized, and an analysis apparatus and an analysis method using the same. With the goal.

  The analytical device according to claim 1 of the present invention has a microchannel structure for transporting a sample solution toward a measurement cell by centrifugal force, and reads for accessing a reaction solution of the sample solution and a reagent in the measurement cell. The measurement cell is formed by extending in the direction in which the centrifugal force acts, and is formed in the measurement cell so as to extend from the outer peripheral position of the measurement cell to the inner peripheral direction. A capillary area is formed, one end of the capillary area on the outer peripheral position side is connected to a side wall positioned in the rotation direction of the measurement cell, and the inner peripheral direction from one end of the capillary area on the outer peripheral position side At least a part of one end extending in the direction is formed apart from the side wall located in the rotation direction of the measurement cell.

The analytical device according to claim 2 of the present invention is characterized in that, in claim 1, the capillary area has a capacity capable of accommodating all the sample liquid held in the measurement cell.
According to a third aspect of the present invention, there is provided the analytical device according to the first aspect, wherein the reagent is carried in an area formed apart from a side wall located in the rotation direction in the capillary area. And

The analysis device according to claim 4 of the present invention is characterized in that, in claim 1, a plurality of the measurement cells are arranged in the circumferential direction.
The analysis device according to claim 5 of the present invention is characterized in that, in claim 4, the plurality of measurement cells are arranged on the same radius.

  In the analysis device according to claim 6 of the present invention, when two or more reagents are carried in the capillary area according to claim 1, a reagent having a higher viscosity is carried at the outer peripheral position than a reagent having a lower viscosity. It is characterized by that.

  The analysis apparatus according to claim 7 of the present invention is an analysis apparatus in which the analysis device according to claim 1 is set, the rotation drive means for rotating the analysis device around an axis, and the rotation drive Control means for transferring to the measurement cell of the analysis device by rotation of the means, and access and analysis of the reaction liquid of the reagent and the sample liquid transferred to the measurement cell of the analysis device by the rotation of the rotation drive means Analyzing means, and the control means transfers the sample liquid to the measurement cell of the analysis device by rotation of the rotation drive means, and then applies the vibration around the axis to the analysis device. The sample liquid in the cell is sucked up into the capillary area, and then the rotation of the analytical device is accelerated to remove the sample liquid sucked up into the capillary area. Characterized by being configured to stir back to the measuring cell.

  An analysis method according to an eighth aspect of the present invention is an analysis method using the analytical device according to the first aspect, wherein the sample solution is used for the analysis by a centrifugal force generated by rotating the analytical device. A first step of transferring the device to a measurement cell of the device; and the measurement device is vibrated by reciprocating left and right about the axis of rotation at a predetermined stop position and swinging the analysis device. The sample liquid transferred to the measurement cell capillary area is sucked into the measurement cell capillary area by capillary force, and then the rotation of the analytical device is accelerated to return the sample liquid sucked to the capillary area to the measurement cell and stir. A second step of rotating the analytical device into a reaction solution of the sample solution and the reagent in the measurement cell at a timing when the measurement cell is positioned at a reading position. And having a third step of reading and access.

  In the analysis method according to claim 9 of the present invention, in claim 8, in the second step, the step of sucking up the sample solution by the capillary force and the step of transferring the sample solution in the capillary area by the centrifugal force in the outer peripheral direction. Is repeatedly performed.

  According to this configuration, it is possible to realize a stirring mechanism that can be measured with a very small amount of sample solution and can be easily downsized.

Embodiments of the analyzing device of the present invention will be described with reference to FIGS.
(Embodiment 1)
1 to 20 show Embodiment 1 of the present invention.

  FIGS. 1A and 1B show a state in which the protective cap 2 of the analysis device 1 is closed and opened. FIG. 2 shows an exploded view with the lower side in FIG. 1 (a) facing upward, and FIG. 3 shows an assembly drawing thereof.

  As shown in FIG. 1 and FIG. 2, this analytical device 1 includes a base substrate 3 having a microchannel structure having a fine concavo-convex shape on one surface, a cover substrate 4 covering the surface of the base substrate 3, It is composed of four parts including a diluent container 5 holding a diluent and a protective cap 2 for preventing sample liquid scattering.

The base substrate 3 and the cover substrate 4 are joined with the diluent container 5 and the like set therein, and the protective cap 2 is attached to the joined substrate.
By covering the openings of several concave portions formed on the upper surface of the base substrate 3 with the cover substrate 4, a plurality of storage areas described later (same as measurement cells described later) and the microchannels connecting the storage areas A flow path having a structure is formed. Necessary ones of the storage areas are loaded with reagents necessary for various analyses. One side of the protective cap 2 is pivotally supported so that it can be opened and closed by engaging with the shafts 6 a and 6 b formed on the base substrate 3 and the cover substrate 4. When the sample liquid to be examined is blood, the gap between the flow paths of the microchannel structure on which the capillary force acts is set to 50 μm to 300 μm.

  The outline of the analysis process using this analytical device 1 is that the sample solution is spotted on the analytical device 1 in which a diluent is set in advance, and at least a part of the sample solution is diluted with the diluent, and then measurement is performed. It is what.

FIG. 4 shows the shape of the diluent container 5.
4A is a plan view, FIG. 4B is a cross-sectional view taken along line AA of FIG. 4A, FIG. 4C is a side view, FIG. 4D is a rear view, and FIG. These are front views seen from the opening 7. The opening 7 is sealed with an aluminum seal 9 as a seal member after filling the interior 5a of the diluent container 5 with the diluent 8 as shown in FIG. 6 (a). On the opposite side of the diluent container 5 from the opening 7, a latch portion 10 is formed. The dilution liquid container 5 is formed between the base substrate 3 and the cover substrate 4 and is set in the dilution liquid container accommodating portion 11 so that the liquid holding position shown in FIG. 6A and the liquid discharge shown in FIG. It is housed in a freely movable position.

FIG. 5 shows the shape of the protective cap 2.
5 (a) is a plan view, FIG. 5 (b) is a cross-sectional view taken along line BB of FIG. 5 (a), FIG. 5 (c) is a side view, FIG. 5 (d) is a rear view, and FIG. These are the front views seen from the opening 2a. As shown in FIG. 6 (a) in the closed state shown in FIG. 1 (a), a locking groove 12 with which the latch portion 10 of the diluent container 5 can be engaged is formed inside the protective cap 2. ing.

  FIG. 6A shows the analysis device 1 before use. In this state, the protective cap 2 is closed, and the latch portion 10 of the diluent container 5 is engaged with the locking groove 12 of the protective cap 2 so as to prevent the diluent container 5 from moving in the arrow J direction. Locked in position. In this state, it is supplied to the user.

  When the protective cap 2 is opened as shown in FIG. 1B against the engagement with the latch portion 10 in FIG. 6A when the sample liquid is spotted, the locking groove of the protective cap 2 As shown in FIG. 6 (b), the bottom portion 2b formed with 12 is elastically deformed, and the engagement between the locking groove 12 of the protective cap 2 and the latch portion 10 of the diluent container 5 is released.

  In this state, the sample solution is spotted on the exposed inlet 13 of the analytical device 1 and the protective cap 2 is closed. At this time, by closing the protective cap 2, the wall surface 12 a forming the locking groove 12 comes into contact with the surface 5 b on the side of the protective cap 2 of the latch portion 10 of the diluent container 5, so that the diluent container 5 is pushed in the direction of arrow J (direction approaching the liquid discharge position). An opening rib 14 as a protruding portion is formed in the diluent container housing portion 11 from the base substrate 3 side, and when the diluent container 5 is pushed in by the protective cap 2, the diluent container 5 is inclined obliquely. The aluminum seal 9 stretched on the sealing surface of the opening 7 collides with the opening rib 14 and is broken as shown in FIG.

  FIG. 7 shows a manufacturing process in which the analysis device 1 is set in the shipping state shown in FIG. First, before closing the protective cap 2, the groove 42 (see FIGS. 2 and 4 (d)) provided in the lower surface of the diluent container 5 and the hole 43 provided in the cover substrate 4 are aligned, At the liquid holding position, the protrusion 42a of the locking jig 44 provided separately from the base substrate 3 or the cover substrate 4 is engaged with the groove 42 of the diluent container 5 through the hole 43 to hold the diluent container 5 with the liquid. Set in a locked state. Then, the protective cap 2 is closed in a state where the pressing jig 46 is inserted from the notch 45 (see FIG. 1) formed on the upper surface of the protective cap 2 and the bottom surface of the protective cap 2 is pressed and elastically deformed. Can be set to the state shown in FIG. 6A by releasing the pressing jig 46.

  In this embodiment, the case where the groove 42 is provided on the lower surface of the diluent container 5 has been described as an example. However, the groove 42 is provided on the upper surface of the diluent container 5, and the base substrate corresponds to the groove 42. 3 can be configured such that a hole 43 is provided in the groove 3 and the protrusion 44 a of the locking jig 44 is engaged with the groove 42.

  Further, the locking groove 12 of the protective cap 2 directly engages the latch portion 10 of the diluent container 5 to lock the diluent container 5 in the liquid holding position. And the latch portion 10 of the diluent container 5 can be indirectly engaged to lock the diluent container 5 in the liquid holding position.

As shown in FIGS. 8 and 9, the analysis device 1 is set on the rotor 101 of the analyzer 100 with the cover substrate 4 facing downward, so that the component analysis of the sample solution can be performed.
A groove 102 is formed on the upper surface of the rotor 101. When the analysis device 1 is set on the rotor 101, the groove 102 is formed on the rotation support 15 and the protective cap 2 formed on the cover substrate 4 of the analysis device 1. The rotation support portion 16 engages with and accommodates the groove 102.

  When the analysis device 1 is set on the rotor 101 and the door 103 of the analyzer is closed before the rotor 101 is rotated, the set analysis device 1 is moved by the movable piece 104 provided on the door 103 side. The position on the rotational axis of the rotor 101 is pressed against the rotor 101 by the biasing force of the spring 105, and the analysis device 1 rotates integrally with the rotor 101 that is rotationally driven by the rotational driving means 106. Reference numeral 107 denotes an axial center during rotation of the rotor 101. The protective cap 2 is attached in order to prevent the sample liquid adhering to the vicinity of the inlet 13 from being scattered outside due to centrifugal force during analysis.

  As a material of the parts constituting the analysis device 1, a resin material having a low material cost and excellent mass productivity is desirable. Since the analysis apparatus 100 analyzes the sample liquid by an optical measurement method for measuring light transmitted through the analysis device 1, the material of the base substrate 3 and the cover substrate 4 may be PC, PMMA, AS, MS, or the like. A synthetic resin having high transparency is desirable.

  Further, as the material of the diluent container 5, since it is necessary to enclose the diluent 8 in the diluent container 5 for a long period of time, a crystalline synthetic resin having a low moisture permeability such as PP and PE is desirable. As a material for the protective cap 2, there is no particular problem as long as it is a material with good moldability, and an inexpensive resin such as PP or PE is desirable.

  The base substrate 3 and the cover substrate 4 are preferably joined by a method that hardly affects the reaction activity of the reagent carried in the storage area. Ultrasonic welding or laser welding is less likely to generate reactive gas or solvent during joining. Etc. are desirable.

  Further, a hydrophilic treatment for increasing the capillary force is applied to the portion where the solution is transferred by the capillary force due to the minute gap between the substrates 3 and 4 by joining the base substrate 3 and the cover substrate 4. Specifically, hydrophilic treatment using a hydrophilic polymer or a surfactant is performed. Here, hydrophilic means that the contact angle with water is less than 90 °, more preferably less than 40 °.

FIG. 10 shows the configuration of the analyzer 100.
The analysis apparatus 100 includes a rotation driving unit 106 for rotating the rotor 101, an optical measurement unit 108 as an analysis unit that accesses and analyzes the reactant in the analysis device 1, and the rotation speed and rotation of the rotor 101. Control means 109 for controlling the direction and measurement timing of the optical measurement unit 108, a calculation unit 110 for processing a signal obtained by the optical measurement unit 108 and calculating a measurement result, and a result obtained by the calculation unit 110 It is comprised with the display part 111 for displaying.

  The rotation driving means 106 not only rotates the analyzing device 1 around the rotation axis 107 at a predetermined rotation speed around the rotation axis 107 via the rotor 101 but also centers on the rotation axis 107 at a predetermined stop position. The analyzing device 1 can be swung by reciprocating left and right in a predetermined amplitude range and cycle.

  The optical measurement unit 108 detects the amount of transmitted light that has passed through the analysis device 1 out of the laser light source 112a that irradiates the measurement cell of the analysis device 1 with laser light and the laser light emitted from the laser light source 112a. The photo detector 113a, the laser light source 112b for irradiating the measurement unit different from the measurement cell of the analysis device 1, and the transmitted light that has passed through the analysis device 1 out of the laser light emitted from the laser light source 112b And a photodetector 113b for detecting the amount of light.

  The analysis device 1 is driven to rotate by the rotor 101, and the sample liquid taken into the inside from the injection port 13 is rotated around the rotation axis 107 located on the inner periphery of the injection port 13. The centrifugal force generated and the capillary force of the capillary channel provided in the analysis device 1 are used to transfer the solution inside the analysis device 1. The channel structure will be described in detail along with the analysis process.

FIG. 11 shows the vicinity of the inlet 13 of the analytical device 1.
FIG. 11A shows an enlarged view of the injection port 13 viewed from the outside of the analytical device 1, and FIG. 11B shows the microchannel structure viewed through the cover substrate 4 from the rotor 101 side. It is.

  The inlet 13 is a gap where a capillary force acts like the guiding part 17 via a guiding part 17 where a capillary force acts in a minute gap δ formed between the base substrate 3 and the cover substrate 4. A capillary cavity 19 having a volume capable of holding a required amount of sample liquid 18 is connected. The cross-sectional shape orthogonal to the flow direction of the guide portion 17 (the DD cross section in FIG. 11B) is not a rectangular shape with the back side vertical, but as shown in FIG. It forms in the inclined surface 20 which becomes narrow gradually toward. The guide portion 17, the capillary cavity 19, and the connection portion are formed with a bent portion 22 that forms a recess 21 in the base substrate 3 and changes the direction of the passage.

  A sample liquid receiving cavity 23 is formed at the tip of the capillary cavity 19 as viewed from the guiding portion 17 so that a gap between which the capillary force does not act is formed. A cavity 24 having one end connected to the sample solution receiving cavity 23 and the other end opened to the atmosphere is formed on a side of a part of the capillary cavity 19, the bent portion 22, and the guiding portion 17.

  With this configuration, when blood is spotted on the inlet 13 as the sample solution 18, the sample solution 18 is taken up to the capillary cavity 19 via the guide portion 17. FIG. 12 shows a state before the analysis device 1 after spotting is set on the rotor 101 and rotated. At this time, as explained in FIG. 6C, the aluminum seal 9 of the diluent container 5 collides with the opening rib 14 and is broken. Reference numerals 25a to 25g, 25h, 25i1, 25i2, and 25j to 25n denote air holes formed in the base substrate 3.

The analysis process will be described together with the configuration of the control means 109 that controls the operation of the rotation drive means 106.
− Step 1 −
The analytical device 1 in which the sample liquid to be inspected is spotted on the inlet 13 holds the sample liquid in the capillary cavity 19 as shown in FIG. 13A, and the aluminum seal 9 of the diluent container 5 is broken. In this state, the rotor 101 is set.

− Step 2 −
When the rotor 101 is rotationally driven in the clockwise direction (C2 direction) after the door 103 is closed, the held sample solution is broken at the position of the bent portion 22, and the sample solution in the guide portion 17 is discharged into the protective cap 2. Then, the sample liquid 18 in the capillary cavity 19 flows into the sample liquid receiving cavity 23 and is held as shown in FIG.

  The diluent 8 that has flowed out of the diluent container 5 flows into the holding cavity 27 via the discharge channel 26. When the dilution liquid 8 flowing into the holding cavity 27 exceeds a predetermined amount, the excess dilution liquid 8 flows into the mixing cavity 29 as shown in FIG. When the inflowing diluent 8 exceeds a predetermined amount, the excess diluent 8 flows into the overflow cavities 36a, 36b, 36c, and 36d via the connection channels 34a and 34b and the overflow channel 38. The diluent 8 that has flowed into the overflow cavities 36a, 36b, 36c is held so as not to flow out of the overflow cavities 36a, 36b, 36c by the capillary force of the backflow prevention flow paths 35a, 35b.

  Here, the diluent 8 is a solution having a prescribed absorbance in a specific wavelength region, and the absorbance of the diluent 8 is measured while the diluent 8 flowing into the mixing cavity 29 stays in the mixing cavity 29 ( Primary metering). Specifically, when the analyzing device 1 is rotationally driven in the clockwise direction (C2 direction) and the mixing cavity 29 containing the diluent 8 passes between the laser light source 112b and the photodetector 113b, the calculation unit 110 The detection value of the photodetector 113b is read.

  The connection channel 34a has a siphon structure having a bent portion in the inner circumferential direction from the outermost peripheral portion of the mixing cavity 29, and when the diluent 8 exceeding the bent portion of the connection channel 34a flows in, the siphon effect As a result, the diluent 8 in the mixing cavity 29 is discharged into the overflow cavities 36a, 36b, 36c. Further, by providing a connecting channel 34b for discharging a diluent exceeding a predetermined amount at an inner peripheral position of the connecting channel 34a, a sample solution receiving cavity is supplied from the mixing cavity 29 when an excessive diluent flows. Inflow to 23 is prevented.

  The diluent 8 staying in the mixing cavity 29 is discharged to the overflow cavities 36a, 36b, 36c as time passes, and the sample liquid receiving cavity 23 and the holding cavity 27 as shown in FIG. A predetermined amount of the sample solution 18 and the diluted solution 8 are held.

  The diluent container 5 has a bottom portion opposite to the opening 7 sealed with the aluminum seal 9 and is formed with an arcuate surface 32 as shown in FIGS. 4 (a) and 4 (b). At the liquid discharge position of the diluent container 5 in the state shown in FIG. 13B, as shown in FIG. 15, the center m of the arc surface 32 is offset by a distance d so as to be closer to the discharge flow path 26 side than the rotation axis 107. Therefore, the diluent 8 that has flowed toward the arc surface 32 is changed to a flow (in the direction of arrow n) from the outside toward the opening 7 along the arc surface 32, so that the diluent container 5 is efficiently discharged from the opening 7 to the diluent container housing 11.

− Step 3 −
Next, when the rotation of the rotor 101 is stopped, the sample liquid 18 is primed into the connection channel 30 having a siphon shape connecting the sample liquid receiving cavity 23 and the mixing cavity 29 as shown in FIG. Similarly, the diluting liquid 8 is also primed into the connection channel 41 having a siphon shape connecting the holding cavity 27 and the mixing cavity 29.

− Step 4 −
When the rotor 101 is driven to rotate counterclockwise (C1 direction), the sample liquid 18 in the sample liquid receiving cavity 23 and the diluted liquid 8 in the holding cavity 27 flow into the mixing cavity 29 as shown in FIG. In the mixing cavity 29, it is centrifuged into a diluted plasma component 18a and a blood cell component 18b. Reference numeral 18c represents a separation interface between the diluted plasma component 18a and the blood cell component 18b. Here, since the sample solution 18 and the diluent 8 once collide with the rib 31 and then flow into the mixing cavity 29, the plasma component in the sample solution 18 and the diluent 8 can be uniformly stirred.

  Then, the absorbance of the diluted plasma component 18a centrifuged in the mixing cavity 29 is measured (secondary photometry). Specifically, when the analysis device 1 is rotationally driven in the counterclockwise direction (C1 direction), the calculation unit is at a timing when the mixing cavity 29 containing the diluted plasma component 18a passes between the laser light source 112b and the photodetector 113b. 110 reads the detection value of the photodetector 113b.

  Here, in this embodiment, the blood that is the sample liquid 18 and the diluent 8 are directly mixed and then the diluted plasma component 18a is extracted and reacted with a reagent to analyze a specific component in the plasma component. However, since the ratio of the plasma component in the blood varies among individuals, the dilution ratio of the plasma component varies greatly when directly mixed. Therefore, when the diluted plasma component 18a is reacted with the reagent, the reaction concentration varies and affects the measurement accuracy. Therefore, in order to correct the dispersion of the dilution ratio when the sample solution 18 and the diluent 8 are mixed, the absorbance before and after mixing with the sample solution is measured using a diluent having a specified absorbance in a specific wavelength region. Since the dilution factor is calculated by measuring at the same location of the mixing cavity 29, the variation in the optical path length of the measurement unit can be eliminated, the measurement of the dilution factor with high accuracy can be performed, and the measurement result in the measurement cell On the other hand, variations in dilution ratio can be corrected, and the measurement accuracy is greatly improved. This correction method is also useful for correcting variation in the dilution factor due to variations in the liquid volume of the sample liquid 18 and the diluent 8.

− Step 5 −
Next, when the rotation of the rotor 101 is stopped, the diluted plasma component 18 a is sucked up into the capillary cavity 33 formed on the wall surface of the mixing cavity 29, and is passed through the capillary channel 37 communicating with the capillary cavity 33 as shown in FIG. As shown in b), it flows into the overflow channel 38, the metering channels 39a, 39b, 39c, 39d, 39e, 39f, and 39g, and the fixed amount is held in the metering channels 39a to 39g.

  FIG. 17A shows a perspective view of the capillary cavity 33 and its periphery. The EE cross section in Fig.17 (a) is shown in FIG.17 (b). The capillary cavity 33 and its periphery will be described in detail.

  The capillary cavity 33 is formed from the bottom 29 b of the mixing cavity 29 toward the inner peripheral side. In other words, the outermost peripheral position of the capillary cavity 33 is formed to extend in the outer peripheral direction from the separation interface 18c between the diluted plasma component 18a and the blood cell component 18b shown in FIG. Thus, by setting the outer peripheral side position of the capillary cavity 33 as described above, the outer peripheral end of the capillary cavity 33 is immersed in the diluted plasma component 18a and the blood cell component 18b separated in the mixing cavity 29. Since the plasma component 18a has a lower viscosity than the blood cell component 18b, the diluted plasma component 18a is preferentially sucked out by the capillary cavity 33, and the capillary channel 37, the overflow channel 38, and the metering channels 39a, 39b. , 39c, 39d, 39e, 39f, and 39g, the diluted plasma component 18a can be transferred toward the measurement cells 40a to 40f and 40g.

  In addition, since the blood cell component 18b is also sucked after the diluted plasma component 18a after the diluted plasma component 18a is sucked out, the path to the middle of the capillary cavity 33 and the capillary channel 37 is replaced with the blood cell component 18b. When the overflow channel 38 and the metering channels 39a to 39g are filled with the diluted plasma component 18a, the transfer of the liquid in the capillary channel 37 and the capillary cavity 33 is stopped. The blood cell component 18b is not mixed into the flow paths 39a to 39g.

Therefore, since the liquid feeding loss can be minimized as compared with the conventional configuration, the amount of the sample liquid necessary for the measurement can be reduced.
-Step 6-
Further, when the rotor 101 is rotationally driven in the counterclockwise direction (C1 direction), as shown in FIG. 18A, the diluted plasma component 18a held in the measurement flow paths 39a to 39g is released into the atmosphere. It breaks at the positions of the bent portions 49a, 49b, 49c, 49d, 49e, 49f, and 49g, which are connected to the cavity 48, and flows into the measurement cells 40a to 40f and 40g. Here, the same amount of diluted plasma component 18a flows into each of the measurement cells 40a to 40f.

  At this time, the diluted plasma component 18a in the overflow channel 38 flows into the overflow cavities 36c and 36a via the overflow cavity 36d and the backflow prevention passage 35b. At this time, the sample liquid in the mixing cavity 29 flows into the overflow cavities 36a and 36c via the siphon-shaped connection channel 34a and the overflow cavity 36b.

  The shape of the measurement cells 40a to 40f and 40g is a shape extended in the direction in which the centrifugal force acts. Specifically, the width in the circumferential direction of the analysis device 1 from the center of rotation of the analysis device 1 toward the outermost periphery. It is thin. Since the bottoms on the outer peripheral side of the plurality of measurement cells 40a to 40f and 40g are arranged on the same radius of the analyzing device 1, the laser light source 112a having the same wavelength is used to measure the plurality of measurement cells 40a to 40f and 40g. In addition, it is not necessary to arrange a plurality of photodetectors 113a corresponding to them and different radial distances, so that the cost of the apparatus can be reduced and measurement can be performed using a plurality of different wavelengths in the same measurement cell. Measurement sensitivity can be improved by selecting an optimum wavelength according to the concentration.

  Further, inside each of the measurement cells 40a, 40b, 40d to 40f, as shown in FIGS. 18A, 18B, and 19, the capillary area 47a extends from the outer peripheral position of the measurement cell in the inner peripheral direction. 47b, 47d, 47e, 47f are formed as follows. One end S1 of the capillary areas 47a to 47f on the outer peripheral side is connected to a side wall located in the rotation direction of the measurement cells 40a to 40f. Furthermore, one end S2 on the inner peripheral side of the capillary areas 47a to 47f is formed away from the side wall located in the rotation direction of the measurement cells 40a to 40f. The FF cross section in FIG. 19 is shown to Fig.20 (a). A GG cross section in FIG. 19 is shown in FIG. The one end S1 of the capillary areas 47b to 47f is also formed in the same manner as the capillary area 47a.

  The capillary area 47c provided in the measurement cell 40c has one end S2 on the inner peripheral side divided into two, and any one end S2 is formed away from the side wall located in the rotation direction of the measurement cells 40a to 40f. Has been. A KK cross section in FIG. 19 is shown in FIG.

  Note that the capillary area as seen in the measurement cells 40a to 40f is not formed in the measurement cell 40g. The gaps between the capillary areas 47a to 47f and 47g and the cover substrate 4 are 200 μm to 300 μm. Further, the surface of the cover substrate 4 facing the capillary areas 47a to 47f and 47g is subjected to a hydrophilic treatment.

  The capacity of the capillary area 47a that can be sucked is formed to be smaller than the capacity that can accommodate all the diluted plasma component 18a as the sample liquid held in the measurement cell 40a. Similarly, the capillary areas 47b and 47d to 47f are formed to have a capacity smaller than the capacity capable of accommodating all the sample liquids held in the respective measurement cells 40b and 40d to 40f. For the capillary areas 47c1 and 47c2 of the measurement cell 40c, the sum of the capacity that can be sucked up by the capillary area 47c1 and the capacity that can be sucked up by the capillary area 47c2 is a capacity that can accommodate all the sample liquid held in the measurement cell 40c. Is formed.

  Further, as shown in FIG. 19, the capillary areas 47a, 47b, 47c1, 47c2, 47d, 47e, and 47f carry a reagent T1 that reacts with the sample solution. No reagent is provided in the measurement cell 40g. The reagent T1 carried in the capillary areas 47a, 47b, 47c1, 47c2, 47d to 47f differs depending on the specific component to be analyzed, and the easily soluble reagent is carried on the capillary areas 47a, 47b, 47d to 47f. The area 47c is loaded with a reagent that is hardly soluble.

-Step 7-
Next, the analysis device 1 is vibrated by swinging the analysis device 1 by reciprocating left and right at a predetermined amplitude range and a predetermined period around the rotation axis 107 at a predetermined stop position. The sample solution or the mixed solution of the reagent and the sample solution transferred to each measurement cell 40a to 40f is sucked up into the capillary areas 47a to 47f by the capillary force as shown in FIG. Dissolution is initiated, and the reaction between the specific components contained in the diluted plasma component 18a and the reagent is initiated.

  When the mixed solution is sucked into the capillary areas 47a to 47f, the portion of the one end S1 connected to the side wall located in the rotation direction of the measurement cells 40a to 40f is simply reduced or stopped. Even when the mixed solution is sucked up by capillary force, the mixed solution cannot be sucked up to the one end S2 that is separated from the side wall located in the rotation direction of the measurement cells 40a to 40f. By oscillating and vibrating the analyzing device 1, the mixed solution could be reliably sucked up to the one end S2 of the capillary areas 47a to 47f by the capillary force.

  In addition, since the reagent T1 is carried on one end S2 on the inner circumferential side formed away from the side wall located in the rotation direction of the capillary areas 47a to 47f, the areas of the measurement cells 40a to 40f are increased. Even without taking a sufficiently large amount of the reagent to be dropped, there is no deviation of the reagent concentration in the flow channel as seen in the conventional FIGS. 26 (a) and 26 (b), and the reagent T1 is easily dissolved and uniform. A color reaction can be expected, and as a result, even with a small amount of sample solution, there is little variation in measurement results, and an improvement in analysis accuracy can be expected.

− Step 8 −
As shown in FIG. 18B, from the state in which the sample solution or the mixed solution of the reagent and the sample solution is sucked up into the capillary areas 47a to 47f, the rotation of the analysis device 1 is accelerated to make the analysis device 1 When rotated counterclockwise (C1 direction) or clockwise (C2 direction), as shown in FIG. 18 (a), the liquid held in the capillary areas 47a to 47f is measured by the centrifugal force to measure cells 40a to 40f. The reagent T1 and the diluted plasma component 18a are agitated by being transferred to the outer peripheral side.

  Here, since the agitation of the reagent and the diluted plasma component 18a is promoted by repeating the operations of the step 7 and the step 8, the agitation can be surely performed in a shorter time than the agitation only by diffusion. It becomes.

− Step 9−
When the analysis device 1 is rotationally driven counterclockwise (C1 direction) or clockwise (C2 direction), each of the measurement cells 40a to 40f and 40g passes through between the laser light source 112a and the photodetector 113a, and the arithmetic unit 110 reads the detection value of the photo detector 113a and corrects it with the result of the primary photometry and the secondary photometry to calculate the density of the specific component.

The measurement result in the measurement cell 40g is used as reference data for the measurement cells 40a to 40f in the calculation process in the calculation unit 110.
As described above, since the diluent container 5 can be opened by opening and closing the protective cap 2 when the user collects the sample solution, and the diluent can be transferred into the analysis device 1, the analyzer can be simplified. The cost can be reduced, and the operability for the user can be improved.

  Further, the diluent container 5 sealed with the aluminum seal 9 as the sealing member is used, and the diluent container 5 is opened by breaking the aluminum seal 9 with the opening rib 14 as the protruding portion. The analysis accuracy can be improved without the dilution liquid evaporating and decreasing.

  6A, the latch portion 10 of the diluent container 5 is engaged with the locking groove 12 of the closed protective cap 2, and the diluent container 5 is engaged. Is held at the liquid holding position so that it does not move in the direction of arrow J, so that the diluent container 5 is configured to be movable in the diluent container housing portion 11 by opening and closing the protective cap 2. During the period until the user opens the protective cap 2 and uses it, the position of the diluent container 5 in the diluent container housing 11 is locked to the liquid holding position, so that the user can transport the container before use. There is no case where the diluent container 5 is accidentally opened and the diluent is spilled.

  In addition, the width (circumferential dimension) of each of the measurement cells 40a to 40f and 40g formed so as to extend in the centrifugal direction (radial direction) of the analytical device 1 is set to a minimum dimension that can be detected by the optical measurement unit 108. The liquid level height of the liquid held in the measurement cells 40a to 40f and 40g during rotation is defined as a radial position where the optical measurement unit 108 can detect the liquid level height, that is, the liquid level height that satisfies the laser irradiation area Therefore, since it is possible to measure with the minimum amount of liquid, it is possible to measure with a smaller amount of sample liquid than in the case of Patent Document 1.

  As described above, the measurement cells 40a to 40f are formed to extend in the direction in which the centrifugal force acts, and extend from the outer peripheral position of the measurement cells 40a to 40f in the inner peripheral direction on at least one side wall of the rotation direction. Since the capillary areas 47a to 47f are formed and the steps 7 to 9 are executed, the capillary tube areas 47a to 47f are configured to include the inflow path 114, the measurement cell 115, and the flow path 117 for stirring the sample solution and the reagent as seen in Patent Document 1. Even if the U-shaped stirring mechanism is not provided, a sufficient stirring effect can be obtained, and the analysis device can be downsized.

  In addition, when the reagent T1 is supported near the bottom of the measurement cells 40a to 40f, if the stirring and dissolution becomes insufficient, the remaining reagent T1 causes uneven concentration of the reaction solution. Although it will have an influence, improvement in reliability can be expected by adopting a configuration in which the reagent T1 is not carried near the bottom of the measurement cells 40a to 40f as described above.

(Embodiment 2)
FIG. 21 shows a second embodiment of the present invention.
In the first embodiment, the reagent T1 is supported on the capillary areas 47a to 47f. However, as shown in FIG. 21, a plurality of reagents T1 and T2 can be supported on the capillary areas 47a to 47f.

  In such a case, the capillaries areas 47a, 47b, 47d to 47f have a capacity smaller than the capacity that can accommodate all the sample liquids held in the measurement cells 40a, 40b, 40d to 40f. It is preferable to carry a difficult-to-dissolve reagent at the outer peripheral position than a readily soluble reagent.

  Specifically, in the case where the reagent T1 is a pigment that has a viscosity of 1.10 mPa · s and is relatively soluble, and the reagent T2 is a protein that has a viscosity of 3.02 mPa · s and is less soluble than the reagent T1, the capillary area 47a , 47b, 47d to 47f, even if the capacity of the sample cells held in the respective measurement cells 40a, 40b, 40d to 40f cannot be accommodated, the outer peripheral position of the reagent T1 that is difficult to dissolve the reagent T2 that is difficult to dissolve. It was confirmed by experiments that the reagents T1 and T2 were sufficiently dissolved by the stirring and mixing operation. When the reagent T1 is a protein having a viscosity of 3.02 mPa · s and the reagent T2 is a dye having a viscosity of 1.10 mPa · s, the reagent having a low viscosity flows to the outer peripheral side and the stirring and mixing operation is performed. Since the amount of liquid sucked into the capillary area is reduced, only the supernatant sample liquid that does not contain a low-viscosity reagent is sucked up and the stirring effect is small, so the protein of reagent T1 Was undissolved.

  In the specific example of the second embodiment, the capacities of the capillary areas 47a, 47b, 47d to 47f for supporting a plurality of reagents contain all the sample liquids held in the respective measurement cells 40a, 40b, 40d to 40f. The case where the capacity is not possible has been described, but when a plurality of reagents are arranged in a single capillary area, the configuration in which a highly viscous reagent is supported at the outer peripheral position has a capacity of the capillary areas 47a, 47b, 47d to 47f. It is also effective in the case of a capacity that can accommodate all the sample liquids held in the measurement cells 40a, 40b, 40d to 40f.

(Embodiment 3)
22 and 23 show a third embodiment of the present invention.
In the first embodiment, all of the one ends S2 of the capillary areas 47a to 47f provided in the measurement cells 40a to 40f are from the side walls located in the rotation direction of the measurement cells 40a to 40f as shown in FIG. In the third embodiment, as shown in FIG. 22, at least a part of the one end S2 of the capillary areas 47a to 47f provided in the measurement cells 40a to 40f is measured. It is formed apart from the side wall located in the rotation direction of the cells 40a to 40f. Specifically, the one end S1 of the outermost periphery of the capillary area 47a of the measurement cell 40a is connected to the side wall located in the rotation direction of the measurement cell 40a as shown in FIG. The one end S2 of the capillary area 47a extended in the inner peripheral direction from the one end S1 on the outer peripheral position side of the area 47a is the rotation direction of the measuring cell 40a as shown in FIG. The other area of the one end S2 of the capillary area 47a is formed away from the side wall located in the rotation direction of the measurement cells 40a to 40f as shown in FIG. 23 (b). Yes. The reagent T1 is carried in an area formed away from the side wall located in the rotation direction of the measurement cell 40a at the one end S2 of the capillary area 47a.

  Thus, by connecting a part of the one end S2 of the capillary area 47a to the side wall located in the rotation direction of the measuring cell 40a, the capillary force that the capillary area 47a sucks up the mixed solution is compared with the case of the first embodiment. It can be expected to be sufficiently mixed and stirred. The capillary areas 40b to 40f are the same, and FIGS. 23 (d) and 23 (e) show the KK cross section and the LL cross section of the end S2 in the capillary areas 47c1 and 47c2 of the measurement cell 40c.

  In FIGS. 22 and 23, the case where each capillary area 47a to 47f carries a single reagent T1 has been described. In the third embodiment, the reagent is placed in the capillary areas 47a to 47f as in the second embodiment. The same can be done when T1 and T2 are supported.

  The present invention is useful as a stirring means for an analytical device used for component analysis of a liquid collected from a living organism.

FIG. 3 is an external perspective view of the analytical device according to the embodiment of the present invention with the protective cap closed and opened. Exploded perspective view of the analysis device of the same embodiment A perspective view of the analytical device with the protective cap closed as seen from the back Explanatory drawing of the diluent container of the embodiment Explanatory drawing of the protective cap of the embodiment Sectional drawing of the state in which the protective cap is closed before using the analytical device of the same embodiment, when spotting the sample liquid, and after spotting Cross-sectional view of the process to set to the shipping state Perspective view just before setting the analysis device to the analyzer Sectional view of the analysis device set in the analyzer Configuration diagram of the analyzer of the same embodiment Expansion explanatory drawing of the principal part of the analysis device of the embodiment Sectional view before setting the analysis device to the analyzer and starting rotation Sectional view after setting the analytical device in the analyzer and rotating and then centrifuging Sectional view when the solid component of the sample solution after centrifugation is quantitatively collected and diluted Cross section of the diluent container at the timing when the diluent is released from the rotational axis of the analytical device and the diluent container Cross section of process 4 and process 5 Capillary cavity 33 and its surrounding enlarged perspective view and EE cross section Sectional drawing of process 6 and process 7 FIG. 12 is an enlarged plan view of the measurement cells 40a to 40f. FF sectional view in FIG. 18, GG sectional view, and KK sectional view Enlarged plan view of measurement cells 40a to 40f in Embodiment 2 of the present invention. Enlarged plan view of measurement cells 40a to 40f in Embodiment 3 of the present invention. Sectional drawing of the principal part of the same embodiment A plan view and a cross-sectional view of the analytical device described in Patent Document 1. Plan view of analysis device described in Patent Document 2 Sectional drawing immediately after dripping the reagent 122 to the reaction chamber 119B of patent document 2, and after drying

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Analytical device 2 Protective cap 2a Opening 2b Bottom part 3 Base substrate 4 Cover substrate 5 Diluent container 5a Inside 5b Surface of latch part 10 6a, 6b Shaft 7 Opening 8 Diluent 9 Aluminum seal (seal member)
DESCRIPTION OF SYMBOLS 10 Latch part 11 Diluent container accommodating part 12 Groove for latching 12a Wall surface 13 Inlet 14 Opening rib (protrusion part)
DESCRIPTION OF SYMBOLS 15,16 Rotation support part 17 Guidance part 18 Sample liquid 18a Diluted plasma component 18b Blood cell component 19 Capillary cavity 20 Inclined surface 21 Recessed part 22 Bending part 23 Sample liquid receiving cavity 24 Cavity 25a-25h, 25i1, 25i2, 25j-25n Air hole 26 discharge flow path 27 holding cavity 28 overflow flow path 29 mixing cavity 30 connection flow path 31 rib 32 arc surface 33 capillary cavity 34a connection flow path 34b connection flow path 35a reverse flow prevention path 35b reverse flow prevention path 36a, 36b, 36c, 36d Overflow cavity 37 Capillary channel 38 Overflow channel 39a, 39b, 39c, 39d, 39e, 39f, 39g Metering channel 40a, 40b, 40c, 40d, 40e, 40f, 40g Measurement cell 41 Connection channel 42 Groove 43 Hole 44 Locking jig 44 Projection 45 Notch 46 Pressing jig 47a, 47b, 47c1, 47c2, 47d, 47e, 47f Capillary area 48 Atmospheric release cavity 49a, 49b, 49c, 49d, 49e, 49f, 49g Bending part 100 Analyzing apparatus 101 Rotor 102 Groove 103 Door 104 Movable piece 105 Spring 106 Rotation drive means 107 Rotation axis 108 Optical measurement unit (analysis means)
DESCRIPTION OF SYMBOLS 109 Control means 110 Calculation part 111 Display part 112a, 112b Laser light source 113a, 113b Photodetector T1, T2 Reagent S1 One end by the side of the outer periphery of capillary area 47a-47f S2 One end by the side of the inner periphery of capillary area 47a-47f

Claims (9)

An analytical device having a microchannel structure for transferring a sample solution toward a measurement cell by centrifugal force and used for reading to access a reaction solution of the sample solution and a reagent in the measurement cell,
The measurement cell is formed by extending in the direction in which the centrifugal force works,
A capillary area is formed inside the measurement cell so as to extend in the inner circumferential direction from the outer peripheral position of the measurement cell;
One end of the capillary area on the outer peripheral position side is connected to a side wall located in the rotation direction of the measurement cell, and at least a part of one end extending in the inner peripheral direction from one end on the outer peripheral position side of the capillary area The analysis device is formed so as to be separated from the side wall located in the rotation direction of the measurement cell. The analytical device according to claim 1, wherein the capillary area has a capacity capable of accommodating all the sample liquid held in the measurement cell. The analytical device according to claim 1, wherein the reagent is carried in an area formed apart from a side wall located in the rotation direction in the capillary area. The analysis device according to claim 1, wherein a plurality of the measurement cells are arranged in the circumferential direction. The analysis device according to claim 4, wherein the plurality of measurement cells are arranged on the same radius. The analytical device according to claim 1, wherein when two or more reagents are carried in the capillary area, a reagent having a high viscosity is carried at the outer peripheral position than a reagent having a low viscosity. An analysis apparatus in which the analysis device according to claim 1 is set,
Rotational drive means for rotating the analytical device about an axis;
Control means for transferring to the measurement cell of the analytical device by rotation of the rotation drive means;
An analysis means for accessing and analyzing a reaction solution of the reagent and the sample liquid transferred to the measurement cell of the analytical device by the rotation of the rotation driving means; and the control means,
After the sample liquid is transferred to the measurement cell of the analytical device by rotation of the rotation driving means, the sample liquid of the measurement cell is sucked up to the capillary area while applying vibration around the axis to the analytical device. Then, the analysis apparatus is configured to accelerate the rotation of the analysis device and return the sample liquid sucked into the capillary area to the measurement cell and stir. An analysis method using the analysis device according to claim 1,
A first step of transferring the sample liquid to the measurement cell of the analytical device by centrifugal force generated by rotating the analytical device;
The sample liquid transferred to the measurement cell by vibrating the analysis device by swinging the analysis device by reciprocating left and right around the rotation axis at a predetermined stop position is used. A second step of accelerating the rotation of the analytical device and sucking the sample liquid sucked into the capillary area back to the measurement cell after being sucked into the area by capillary force; and
And a third step of accessing and reading the reaction solution of the sample liquid and the reagent in the measurement cell at a timing when the measurement cell is positioned at the reading position by rotating the analysis device. 9. The analysis method according to claim 8, wherein in the second step, the step of sucking up the sample solution by the capillary force and the step of transferring the sample solution in the capillary area in the outer peripheral direction by the centrifugal force are repeated.

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