JP2007040845A - Reaction vessel and analytical apparatus using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a reaction vessel capable of making a liquid sample minute, and to provide an analytical apparatus capable of being made compact. <P>SOLUTION: The analyzing apparatus and the reaction vessel which has a sidewall and a bottom wall, to which the liquid sample including a specimen and a reagent is dispensed, and which holds a reaction liquid resulting from the reaction of the liquid sample, are provided. In the reaction vessel 15, the horizontal cross-sectional area of the upper section of the side wall is made larger than that of the lower section of the sidewall in order to be made stackable, and a portion or the whole of the bottom wall is made of a metal thin film 15b. The analyzing apparatus comprises a constant-temperature bath 9 for immersing the metal thin film of the reaction vessel in a transparent constant-temperature liquid Lt and for keeping the liquid sample at a prescribed temperature; a light source 6a for irradiating the metal thin film with light through the constant-temperature bath, at an incidence angle not smaller than the critical angle; a light-receiving element 6d for receiving light reflected by the metal thin film; and a control section which calculates a refractive index of the reaction liquid, by using a resonance angle of surface plasmon resonance caused by the light impinging on the metal thin film, and obtains the material concentration of the reaction liquid, based on the relation between the refractive index and absorbance of the reaction liquid at the measured resonance angle and the relation between the absorbance and the material concentration of the reaction liquid. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a reaction vessel and an analyzer using the reaction vessel.

  Conventionally, an analyzer for analyzing a biological sample (specimen) such as blood dispenses and reacts a specimen and a reagent in a reaction container, and the absorbance of the reaction liquid is measured photoelectrically. Analyzing substance concentration. At this time, since the analyzer is excellent in terms of simplification of the measurement mechanism, various types of reaction vessels have been proposed by adopting a method in which photometry is performed while the reaction solution is kept in the reaction vessel (for example, patent documents). 1).

Japanese Patent Publication No. 1-2455

  By the way, in order to measure the light absorbency of a reaction liquid, the conventional reaction container needs to permeate | transmit a light beam with a diameter of 2-3 mm to a reaction liquid. For this reason, the conventional reaction vessel requires at least several tens of μL of the reaction solution, and it is difficult to reduce the size of the sample due to the minute amount of the sample for reducing the physical burden on the patient, and it is also difficult to reduce the size of the analyzer. was there. In addition, since the analyzer uses a large number of reaction containers in view of handling a large number of samples at a time, it requires a wide space for accommodating the large number of reaction containers and is difficult to reduce in size.

  The present invention has been made in view of the above, and an object of the present invention is to provide a reaction vessel capable of reducing the amount of a liquid sample and an analyzer capable of being miniaturized.

  In order to solve the above-described problems and achieve the object, the reaction container according to claim 1 has a side wall and a bottom wall, dispenses a liquid sample containing a specimen and a reagent, and the liquid sample reacts. A reaction vessel for holding the reaction liquid, wherein the horizontal cross-sectional area of the upper side wall is set larger than the horizontal cross-sectional area of the lower side wall and can be stacked, and a part of the bottom wall or The whole is a metal thin film.

  The reaction container according to claim 2 is characterized in that, in the above-mentioned invention, the side wall is provided with a convex portion forming a gap with a side wall of another stacked container.

  In order to solve the above-described problems and achieve the object, an analyzer according to claim 3 is an analyzer that analyzes a reaction liquid by reacting a liquid sample containing a specimen and a reagent held in a container. And immersing the metal thin film of the reaction vessel in a transparent thermostat, and holding the liquid sample at a predetermined temperature, and passing the light through the thermostat at an incident angle greater than a critical angle to the metal thin film. The refractive index of the reaction solution is calculated from the resonance angle of the surface plasmon resonance of the light source irradiated, the light receiving element that receives the light reflected by the metal thin film, and the light irradiated to the metal thin film, and measured in advance. A control unit for determining the substance concentration of the reaction solution from the relationship between the refractive index and the absorbance of the reaction solution at the resonance angle and the relationship between the absorbance and the substance concentration of the reaction solution. To do.

  According to a fourth aspect of the present invention, there is provided the analyzer according to the above invention, further comprising a container supply device that stores a plurality of the reaction containers stacked and supplies them one by one.

  The analyzer according to claim 5 is characterized in that, in the above-mentioned invention, further comprises a discarding device for discarding the reaction container whose measurement has been completed.

  Since the reaction container according to the present invention uses a metal thin film for a part or all of the bottom wall, it can be analyzed even with a small amount of liquid sample by utilizing surface plasmon resonance. Further, the analyzer of the present invention using this reaction vessel includes a constant temperature bath that holds the liquid sample at a predetermined temperature by immersing the thin metal film of the reaction vessel in a transparent constant temperature solution, and the metal thin film via the constant temperature bath. A light source that emits light at an incident angle greater than a critical angle, a light receiving element that receives light reflected by the metal thin film, and a surface plasmon resonance resonance angle of the light irradiated on the metal thin film. Control for calculating the refractive index and determining the substance concentration of the reaction liquid from the relation between the refractive index of the reaction liquid and the absorbance at the resonance angle measured in advance and the relation between the absorbance and the substance concentration of the reaction liquid. And equipped with. For this reason, the analyzer of the present invention can analyze even if the liquid sample containing the sample is in a very small amount. Further, the reaction vessel to be miniaturized can be stacked by setting the horizontal cross-sectional area of the upper side wall to be larger than the horizontal cross-sectional area of the lower side wall. For this reason, since the analyzer of the present invention can be accommodated in a compact manner without taking up space even with a plurality of reaction vessels, there is an effect that downsizing becomes possible.

  Hereinafter, embodiments of a reaction container of the present invention and an analyzer using the reaction container will be described in detail with reference to the drawings. FIG. 1 is a schematic configuration diagram of an automatic analyzer showing an embodiment of the analyzer of the present invention. FIG. 2 is an enlarged view showing a cross section of the photometric unit, the thermostat, and the turntable in the vicinity of the photometric unit of the automatic analyzer shown in FIG. FIG. 3 is a perspective view of the reaction vessel of the present invention used in the automatic analyzer of FIG.

  As shown in FIG. 1, the automatic analyzer 1 includes a sample dispensing unit 3, a reagent dispensing unit 4, a stirring unit 5, a photometric unit 6, a discarding device 7, a container feeding device 8, and the periphery around and below the turntable 2. A thermostatic chamber 9 is disposed and includes a control unit 11 and a storage unit 12.

  The turntable 2 is rotated around the rotating shaft 2b by the driving means 2a, and reaction vessels 15 are arranged at equal intervals along the circumferential direction on the outer edge side. As shown in FIG. 2, the turntable 2 has an opening 2 c in a portion where the reaction vessel 15 is disposed. Here, the turntable 2 rotates by one reaction container 15 every rotation. That is, in the turntable 2 shown in FIG. 1, 16 reaction vessels 15 are arranged in the circumferential direction, but when rotated once, instead of 360 °, 360 ° + one reaction vessel 15 (= 22. 5 °) rotate a lot.

  The sample dispensing unit 3 is a part for dispensing a biological sample (specimen) such as serum into the reaction container 15, and a probe is provided at the tip of the dispensing arm 3a.

  The reagent dispensing unit 4 is a part for dispensing the reagent into the reaction container 15, and a probe provided at the tip of the dispensing arm 4 a dispenses the reagent from the predetermined reagent container to the reaction container 15 sequentially. Here, although the reagent dispensing part 4 is one place in the automatic analyzer 1 shown in FIG. 1, it may be provided at two places or more so that two or more kinds of reagents can be used.

  The agitating unit 5 is a part for agitating a liquid sample containing a specimen and a reagent dispensed in the reaction container 15. For example, the agitating part 5 is held in the reaction container 15 by a sound wave generating element arranged at the lower outer edge of the turntable 2. The liquid sample is stirred in a non-contact manner by sound waves.

  As shown in FIG. 2, the photometry unit 6 includes a light source 6a, a lens 6b, a drive unit 6c, and a light receiving element 6d. The light source 6a irradiates the metal thin film 15b of the reaction vessel 15 with p-polarized light so that the incident angle is greater than the critical angle. The lens 6b collects the light emitted from the light source 6a and irradiates the lower surface of the metal thin film 15b. The drive unit 6c is an actuator that continuously changes the incident angle of light incident on the metal thin film 15b by changing the direction of the light source 6a in an angle range equal to or greater than the critical angle, and sets and changes the incident angle with high accuracy. A possible angle stage or the like is used. The light receiving element 6d receives the light reflected by the metal thin film 15b and outputs the light to the control unit 11 as an optical signal related to absorbance. Here, if the incident angle can be measured, the incident angle may be obtained from the reflection angle of the reflected light received by the light receiving element 6d.

  The discarding device 7 discards the reaction container 15 that has been analyzed and is conveyed by the rotation of the turntable 2.

  As shown in FIGS. 4 and 5, the container supply device 8 includes a storage unit 8 a, arms 8 b and 8 c and a drive unit 8 d that individually drives the arms 8 b and 8 c of the reaction container 15. The arms 8b and 8c are notched in a semicircular shape at the distal end portion according to the side shape of the main body 15a of the reaction vessel 15. The container supply device 8 supports a plurality of reaction vessels 15 accommodated in the accommodating portion 8a by an arm 8b, and moves the arms 8b and 8c in the left-right direction in the drawing by a drive portion 8d, thereby moving the reaction vessel 15 to the turntable 2. One by one is supplied to the openings 2c.

  The thermostatic chamber 9 is formed into a ring shape from an optically transparent material so that the light emitted from the light source 6a of the photometry unit 6 is irradiated onto the metal thin film 15b and the reflected light can be received by the light receiving element 6d. , Is disposed below the opening 2c of the turntable 2. As shown in FIG. 2, the thermostatic chamber 9 is transparent and holds a thermostatic liquid Lt such as cedar oil having a refractive index higher than that of the reaction liquid Lr of sample and reagent and air, and a temperature control device 10. Thus, the constant temperature liquid Lt is maintained at a predetermined temperature, for example, 37 ° C. Accordingly, the light emitted from the light source 6a is refracted when entering the constant temperature liquid Lt, and is applied to the metal thin film 15b of the reaction vessel 15 so as to have an incident angle greater than the critical angle. Further, the liquid sample containing the specimen and the reagent dispensed into the reaction container 15 is also maintained at 37 ° C. For this reason, the automatic analyzer 1 can accurately obtain the resonance angle of the reaction liquid Lr, and thus the substance concentration of the reaction liquid Lr, by the reaction vessel 15.

  The control unit 11 is connected to the sample dispensing unit 3, the reagent dispensing unit 4, the stirring unit 5, the photometric unit 6, the discarding device 7, and the container supply device 8. For example, a microcomputer or the like is used. The control unit 11 controls the operation of each unit of the automatic analyzer 1. At this time, the control unit 11 measures the incident angle θ of the light incident on the metal thin film 15b when the reflectance of the light on the metal thin film 15b is drastically reduced based on the drive signal output to the drive unit 6c. This incident angle θ is the resonance angle θsp when the surface plasmon resonance phenomenon occurs. Further, the control unit 11 calculates the refractive index of the reaction liquid Lr from the measured resonance angle θsp and a known value, and obtains the substance concentration of the reaction liquid Lr from a calibration curve or the like stored in the storage unit 12. The storage unit 12 stores a calibration curve, a calibration table, and the like prepared in advance based on the refractive index and absorbance of the reaction liquid Lr of the sample and reagent dispensed in advance in the reaction container 15.

  As shown in FIG. 3, the reaction vessel 15 has a main body 15a made of an inverted frustoconical member with the top of a cone cut as a side wall, and the bottom surface of the main body 15a is a metal with a thickness of several tens of nm made of gold or silver. The bottom wall is covered with the thin film 15b. The reaction container 15 is a disposable container, and the main body 15a is molded from a synthetic resin in order to be manufactured at a low cost. The reaction container 15 is provided with protrusions 15c at appropriate locations on the outer surface of the main body 15a. The protrusions 15c form a gap between the main bodies 15a of the other reaction vessels 15 so that the main bodies 15a do not come into close contact with each other when stacked. Therefore, the reaction vessel 15 may be a ridge extending in the circumferential direction instead of the protrusion 15c. When the reaction vessel 15 is arranged in the opening 2c of the turntable 2, the metal thin film 15b serving as the bottom wall is immersed in the constant temperature liquid Lt of the constant temperature bath 9, and the transparent constant temperature liquid Lt and the metal thin film 15b are measured by the photometric unit. The surface plasmon resonance sensor is configured in cooperation with the light source 6a and the light receiving element 6d.

  In the automatic analyzer 1 configured as described above, the sample dispensing unit 3 sequentially dispenses the sample into the reaction container 15 conveyed by the rotation of the turntable 2. The reaction container 15 into which the sample has been dispensed is conveyed to the vicinity of the reagent dispensing unit 4 by the rotation of the turntable 2, and the reagent is dispensed from the probe of the reagent dispensing unit 4. Then, in the reaction container 15 into which the reagent has been dispensed, the reagent and the sample are stirred and reacted in the stirring unit 5, and the reaction solution is measured by the photometric unit 6 to determine the incident angle θ and the refractive index of the light transmitting unit. Based on the analysis, the substance concentration in the reaction solution is analyzed. Then, the automatic analyzer 1 further conveys the reaction vessel 15 by the rotation of the turntable 2, discards the reaction vessel 15 that has been analyzed in the discarding device 7, and then supplies a new reaction vessel 15 by the vessel supply device 8. Then, a new sample is analyzed again.

  At this time, as shown in FIG. 6, the container supply device 8 is stopped until the reaction container 15 shown in FIG. 4 is conveyed to the sample dispensing unit 3 side by the rotation of the turntable 2. Then, when the opening 2c in which the reaction vessel 15 is not arranged is moved by the rotation of the turntable 2, the vessel supply device 8 moves the lower arm 8c to the right as shown in FIG. Thereby, the container supply apparatus 8 drops the lowermost reaction container 15 and arranges it in the opening 2c, and supports the second and higher reaction containers 15 by the upper arm 8b.

  Next, as shown in FIG. 8, the container supply device 8 moves the lower arm 8c to the left to return it to its original position. At this time, the reaction container 15 arranged in the opening 2 c is conveyed to the sample dispensing unit 3 side by the rotation of the turntable 2. Next, the container supply device 8 moves the upper arm 8b to the right as shown in FIG. Thereby, the container supply device 8 moves the plurality of reaction vessels 15 supported by the upper arm 8b to the lower arm 8c.

  Thereafter, the container supply device 8 moves the upper arm 8b to the left and returns it to the original position, and supports the plurality of reaction containers 15 by the upper and lower arms 8b and 8c as shown in FIG. . Subsequently, by repeating the same operation, the container supply device 8 supplies the reaction containers 15 accommodated in the accommodating portion 8a one by one to the opening 2c of the turntable 2.

  At this time, the reaction vessel 15 has a bottom wall by covering the bottom surface of the main body 15a with the metal thin film 15b. Further, the automatic analyzer 1 continuously immerses the incident angle θ at an angle equal to or greater than the critical angle by immersing the metal thin film 15b of the reaction vessel 15 in the constant temperature liquid Lt of the constant temperature bath 9 and driving the light source 6a by the driving unit 6c. While changing, the p-polarized light is irradiated onto the metal thin film 15b from below. For this reason, when the reaction vessel 15 is disposed on the turntable 2 of the automatic analyzer 1, the light is irradiated from the light source 6 a to the metal thin film 15 b at an incident angle θ greater than the critical angle when passing through the photometric unit 6. Then, an evanescent wave is generated in the reaction solution near the metal thin film 15b, and a surface plasmon wave is excited on the lower surface of the metal thin film 15b by the light tunnel effect.

At this time, if the wave numbers of the evanescent wave and the surface plasmon wave are equal, a surface plasmon resonance phenomenon occurs, and the energy of the irradiation light is used as the excitation energy of the surface plasmon wave. For this reason, when the surface plasmon resonance phenomenon occurs, as shown in FIG. 11, the reflectance of light in the metal thin film 15b is remarkably reduced at a specific incident angle where resonance occurs, that is, the resonance angle θsp. At this time, the resonance angle θsp is given by the following equation, where ns is the refractive index of the reaction liquid Lr, nt is the refractive index of the constant temperature liquid Lt, and ε is the dielectric constant of the metal thin film 15b.
θsp = sin ⁻¹ [1 / nt {(ε × ns ² ) / (ε + ns ² )} ^1/2 ] (1)

  Accordingly, when the resonance angle θsp is measured, since the dielectric constant ε of the metal thin film 15b and the refractive index np of the constant temperature liquid Lt are known values, the refractive index ns of the reaction liquid Lr can be calculated from the equation (1). . At this time, the refractive index ns of the reaction liquid Lr can be defined by the dielectric constant of the medium from Maxwell's equation, and the correlation between the progress of the biochemical reaction by the biological material and the dielectric constant of the medium, and thus the substance concentration of the reaction liquid Lr. There is. Therefore, the relationship between the refractive index ns of the reaction solution Lr and the absorbance of the reaction solution Lr at the resonance angle θsp and the relationship between the absorbance and the substance concentration of the reaction solution Lr are measured in advance, and a calibration curve or a calibration table is obtained for each measurement target. The data is stored in the storage unit 12. In this way, the automatic analyzer 1 can control the substance of the reaction liquid Lr in the control unit 11 based on the refractive index ns of the reaction liquid Lr at the resonance angle θsp, even for a very small amount of reaction liquid of several μL or less. The concentration can be determined. However, the automatic analyzer 1 stores the calibration curve of the absorbance of the reaction solution Lr and the resonance angle θsp in advance in the storage unit 12 for each measurement target, and the control unit 11 uses the reaction solution Lr based on the resonance angle θsp. The absorbance may be determined.

  Thus, when calculating | requiring the substance concentration of a reaction liquid using surface plasmon resonance, since the reaction liquid Lr should just be a slight quantity of several microliters or less, the main body 15a and hence the reaction container 15 can be made small, Since the reaction containers 15 can be stacked, the automatic analyzer 1 can be downsized. In addition, since the reaction vessel 15 has a reaction solution of several μL or less, the amount of reagent used may be as small as several μL or less, so the running cost of the automatic analyzer 1 can be reduced.

  The reaction vessel 15 can measure the resonance angle θsp of the reaction liquid Lr while immersing the metal thin film 15b serving as the bottom wall in the constant temperature liquid Lt and rotating the turntable 2. Therefore, when the reaction vessel 15 is used, the automatic analyzer 1 can continuously measure the resonance angle θsp without stopping the turntable 2, so that the sample can be processed at a high speed, and therefore the same as in the prior art. Many specimens can be processed in time.

  Here, like the reaction vessel 17 shown in FIG. 12, the reaction vessel of the present invention has a part of the bottom wall 17b connected to the inverted frustoconical side wall 17a, that is, an opening 17c in the center, A part of the bottom wall 17b may be used as the metal foil film 17d by covering the entire lower surface of the bottom wall 17b with the metal foil film 17d. If it does in this way, the reaction container 17 can protect the metal foil film | membrane 17d with the bottom wall 17b. The side wall 17a is provided with a protrusion 17e at an appropriate place.

  Further, the reaction container of the present invention does not necessarily have a wall inverted frustum shape on the side, but has an inverted polygonal frustum shape, for example, a side wall 18a having an inverted quadrangular frustum shape like the reaction container 18 shown in FIG. The bottom surface of the side wall 18a may be covered with a metal thin film 18b made of gold or silver and having a thickness of several tens of nanometers to form a bottom wall. At this time, the side wall 18a is provided with a protrusion 18c at an appropriate location. Further, like the reaction vessel 17, the reaction vessel 18 may have a part of the bottom wall made of a metal thin film.

It is a schematic block diagram of the automatic analyzer which shows embodiment of the analyzer of this invention. FIG. 2 is an enlarged view showing a cross section of a photometric unit, a thermostat, and a turntable near the photometric unit of the automatic analyzer of FIG. 1. It is a perspective view of the reaction container of this invention used with the automatic analyzer of FIG. It is the front view which showed the container supply apparatus with the turntable which has arrange | positioned the reaction container. It is the top view which showed the container supply apparatus with the reaction container. It is a figure which shows the change by the action | operation of the container supply apparatus shown in FIG. 4, and a turntable, and is a front view which shows the 1st state after changing from the position shown in FIG. It is a figure which shows the change by the action | operation of the container supply apparatus shown in FIG. 4, and a turntable, and is a front view which shows the 2nd state after changing from the position shown in FIG. It is a figure which shows the change by the action | operation of the container supply apparatus shown in FIG. 4, and a turntable, and is a front view which shows the 3rd state after changing from the position shown in FIG. It is a figure which shows the change by the action | operation of the container supply apparatus shown in FIG. 4, and a turntable, and is a front view which shows the 4th state after changing from the position shown in FIG. It is a figure which shows the change by the action | operation of the container supply apparatus shown in FIG. 4, and a turntable, and is a front view which shows the 5th state after changing from the position shown in FIG. It is a characteristic view of the reflectance with respect to the incident angle showing an example in which the reflectance of light is significantly reduced at the resonance angle. It is sectional drawing which shows the 1st modification of reaction container. It is a perspective view which shows the 2nd modification of reaction container.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Automatic analyzer 2 Turntable 2c Opening 3 Sample dispensing part 4 Reagent dispensing part 5 Stirring part 6 Photometry part 6a Light source 6b Lens 6c Drive part 6d Light receiving element 7 Discarding device 8 Container supply apparatus 9 Constant temperature bath 10 Temperature control apparatus 11 Control unit 12 Storage unit 15 Reaction vessel 15a Body 15b Metal thin film 15c Protrusion 17 Reaction vessel 17a Side wall 17b Bottom wall 17c Opening 17d Metal foil film 17e Protrusion 18 Reaction vessel 18a Side wall 18b Metal thin film 18c Protrusion

Claims (5)

A reaction container having a side wall and a bottom wall, in which a liquid sample containing a specimen and a reagent is dispensed, and holds a reaction liquid reacted with the liquid sample,
The horizontal cross-sectional area of the upper part of the side wall is set larger than the horizontal cross-sectional area of the lower part of the side wall to enable stacking, and a part or all of the bottom wall is a metal thin film. container.   The reaction container according to claim 1, wherein the side wall is provided with a convex portion on the outer surface to form a gap with a side wall of another stacked container. An analyzer for reacting a liquid sample containing a specimen and a reagent held in a container and analyzing the reaction solution,
A constant temperature bath for immersing the metal thin film of the reaction container according to claim 1 or 2 in a transparent constant temperature liquid and holding the liquid sample at a predetermined temperature;
A light source that irradiates the metal thin film with an incident angle greater than a critical angle through the thermostat; and
A light receiving element for receiving light reflected by the metal thin film;
The refractive index of the reaction solution is calculated from the resonance angle of surface plasmon resonance of light irradiated on the metal thin film, and the relationship between the refractive index and the absorbance of the reaction solution at the resonance angle measured in advance and the absorbance. And a control unit for obtaining the substance concentration of the reaction liquid from the relationship between the substance concentration of the reaction liquid,
An analyzer characterized by comprising:   The analyzer according to claim 3, further comprising a container supply device that stores a plurality of the reaction containers stacked and supplies them one by one.   The analyzer according to claim 3 or 4, further comprising a discarding device for discarding the reaction container after measurement.

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