JP3153676B2 - Sample mixing device and fluorescence detection device using the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sample mixing device in a fluorescence detector or the like used for measuring characteristics of a fluorescent substance or the like in the fields of chemistry, medicine or analytical chemistry.

[0002]

2. Description of the Related Art In the fields of chemistry, medicine, analytical chemistry and the like, the development of fluorescence detectors and the like for measuring characteristics of fluorescent substances and the like has been progressing. When measuring a plurality of samples using a fluorescence detector or the like, as generally called batch processing,
After the measurement of one sample is completed, the reaction container, which is a sample measurement unit of the fluorescence detection device, is removed from the holder, and after cleaning, the reaction container is re-attached to the holder and prepared for the analysis of the next sample. Also,
For the purpose of accelerating the reaction between the sample and the reagent, a stirring member for uniformly mixing the sample and the reagent solution is provided inside the reaction vessel, and a driving mechanism for driving the stirring member is used for the reaction. It is provided outside the container. Further, since the fluorescence intensity varies due to a temperature change in summer or winter, constant temperature water or the like is circulated from a constant temperature water tank to the reaction vessel to keep the temperature constant. In addition, since the amount of a sample to be measured at one time is as small as several millimeters at most, and the reaction container is also small, the stirrer that can be mounted inside the reaction container must be small. It is not possible to use one with a simple structure. Therefore, a stirring member (magnetic stirrer) provided independently inside the reaction vessel and a magnetic drive device (magnetic stirrer) provided at the lower part of the reaction vessel and rotating the stirring member by magnetic force are used. The device is used. The configuration of the conventional sample mixing apparatus as described above is well known or obvious, and therefore is not shown.

[0003]

When a measurement is performed using a conventional sample mixing apparatus, the reaction vessel must be removed from the holder for each measurement of a sample, and washing must be performed. Was virtually impossible to do.
In addition, since the reaction vessel itself is small, a strong (large) magnetic drive device having a strong magnetic force cannot be used, and the distance between the stirring member inside the reaction vessel and the magnetic drive device is longer than a predetermined distance due to an installation error or the like. When the length of the stirrer becomes longer, the strength of the magnetic field acting on the stirrer becomes weaker, and there is a possibility that the rotation of the stirrer may be defective. Furthermore, when the temperature is kept constant, since the constant temperature water is circulated from the constant temperature water tank to the hollow reaction vessel holder, the apparatus becomes large, the structure becomes complicated, and maintenance is required.

Further, since the fluorescence detection device is used for detecting the main component of a stimulant such as methamphetamine in the fields of medical care, police, customs, etc., it is required that the device be small and easy to handle. strong.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and enables continuous measurement of a plurality of samples, no agitation failure, easy maintenance, and small sample mixing. It is an object of the present invention to provide an apparatus and a fluorescence detection apparatus using the same.

[0006]

According to the present invention, there is provided a sample measuring apparatus comprising: a reaction vessel having a rectangular cross section having an inlet and an air vent hole at an upper part and a discharge nozzle at a lower part; And a stirring member provided near the bottom thereof and rotated by a magnetic force from the outside of the reaction vessel, and a magnetic field for rotating the stirring member provided outside the reaction vessel and near the stirring member. The reaction vessel , comprising: a magnetic drive device for adjusting the relative distance between the stirring member and the magnetic drive device.
A Peltier element for temperature control provided in the vicinity of said anti
Container, magnetic drive, adjustment mechanism and Peltier element
Aluminum with black anodized surface for holding
It is characterized by having a holder made of nickel .

In addition, a fluorescence detection device according to the present invention has a reaction vessel having a rectangular cross section having an inlet and an air vent hole at an upper part, and a discharge nozzle at a lower part, and the inside and the inside of the reaction vessel. A stirring member provided near the bottom and rotated by a magnetic force from outside the reaction vessel; and a magnetic drive provided outside the reaction vessel and near the stirring member to generate a magnetic field for rotating the stirring member. Equipment and
Comprising an adjusting mechanism for adjusting the relative distance between the stirring member and the magnetic drive device and a fluorescence detection device for the provided in the vicinity of the reaction vessel for detecting the fluorescence of a sample in the reaction vessel For temperature control in the vicinity of the reaction vessel
Of the reaction vessel, a magnetic drive device,
Black surface to hold adjustment mechanism and Peltier element
With anodized aluminum holder
It is characterized by having .

[0008]

[0009]

In the sample mixing apparatus and the fluorescence detection apparatus using the same according to the present invention, an injection port is provided at the top of the reaction vessel and a discharge nozzle is provided at the bottom, and the cleaning liquid is injected from the injection port. By rotating the stirring member to wash the inside of the reaction vessel and discharging the waste liquid from the discharge nozzle, the inside of the reaction vessel is automatically washed, so that a plurality of samples are continuously measured. It is possible. Further, by adjusting the relative distance between the stirring member and the magnetic driving device for driving the stirring member to a regular distance by the adjusting mechanism, the stirring member is surely rotated at a predetermined rotation speed. Further, since the Peltier element is provided to keep the temperature of the reaction vessel constant, the sample can be measured without being affected by the surrounding temperature conditions. Furthermore, since the holder is made of aluminum having high thermal conductivity, the time required for temperature adjustment is reduced. Further, since the surface of the holder is subjected to black alumite treatment, light scattering during fluorescence detection is reduced.

[0010]

【Example】

<First Embodiment> A sample mixing apparatus according to the present invention and a fluorescence detection apparatus using the same will be described with reference to FIGS. 1, 2 and 3 showing preferred first embodiments. (A) in FIG.
Is a front view showing the configuration of the sample mixing device or the fluorescence detection device,
(B) is a side view thereof. FIG. 2 is a block diagram illustrating a schematic configuration of a sample mixing device or a fluorescence detection device configured to continuously measure a plurality of samples. FIG. 3 is a perspective view showing a detailed configuration of the reaction vessel in FIG. 1 and FIG.

In FIG. 1, a sample mixing apparatus or a fluorescence detecting apparatus 5 of the present invention comprises an aluminum holder 10, a reaction vessel 11 fixed to the holder 10, and an adjusting mechanism (adjusting screw) provided on the holder 10. Reaction vessel 1 through 12
A magnetic driving device (magnetic stirrer) 7 provided outside and at the bottom of the reactor 1; a stirring member (magnetic stir bar) 17 provided inside the reaction vessel 11 and near the bottom; , A fluorescence detection element (or another detection element) 13 or the like. As shown in FIG. 3, an inlet 11c and an air vent hole 11d are provided on the upper surface 11a of the reaction vessel 11, and a discharge nozzle 11b is provided at the bottom. Reaction vessel 11
Is a rectangular cell (having a rectangular cross section) of, for example, 10 mm × 10 mm × 45 mm, and the magnetic drive device 7 is, for example, a cylindrical shape having a diameter of 12 mm × a length of 22 mm. FIG.
As shown in (b), the magnetic drive device 7 is fixed so that it can be adjusted up and down by an adjustment mechanism (adjustment screw) 12 provided at the bottom of the holder 10. Thereby, the relative distance to the stirring member 17 provided inside the reaction vessel 11 can be reduced to a maximum of 3 mm. In addition,
Considering the case of performing temperature control to keep the temperature constant,
The surface of the holder 10 is colored black by alumite treatment in consideration of the case where aluminum having good thermal conductivity is used as the material of the holder 10 and that the holder 10 is used as a fluorescence detector.

As shown in FIG. 2, for example, two inlets 11c provided at the upper part of the reaction vessel 11 are respectively connected to syringe type pumps 1 and 2, and a discharge nozzle 11b is connected to a waste liquid storage via a discharge pump 3. It is connected to the container 4. The pump 1 is used, for example, for controlling the injection amount of an antibody solution as a reagent, and the pump 2 is used for controlling the injection amount of a sample solution. The control device 6 is connected to the magnetic drive device 7.

<Second Embodiment> Next, a sample mixing apparatus according to the present invention and a fluorescence detection apparatus using the same will be described with reference to FIGS. 4 and 5 showing preferred second embodiments thereof. .
4A is a front view showing the configuration of the sample mixing device or the fluorescence detection device, and FIG. 4B is a side view thereof. FIG. 5 is a block diagram illustrating a schematic configuration of a sample mixing device or a fluorescence detection device configured to continuously measure a plurality of samples.
Note that members denoted by the same reference numerals as those in the first embodiment are substantially the same, and a description thereof will be omitted.

4 and 5, the holder 10 is provided with a Peltier element 32 and a radiation fin 33.
The Peltier element 32 which is a temperature control device has a size of 23 m.
m × 23 mm, and is controlled with the power supply current kept constant. The radiating fins 33 were made of aluminum in order to increase the temperature control efficiency.

<First Experimental Example> Using the sample mixing apparatus or the fluorescence detecting apparatus according to the first and second embodiments, the rotation of the stirring member 17, the discharge of waste liquid from the reaction vessel 11, and the second Measurement was performed on the temperature control of the liquid such as the sample in the reaction vessel 11 in the example of (1).

First, regarding the rotation of the stirring member 17, the rotation speed of the stirring member 17 is set to 8 when the liquid exists inside the reaction vessel 11 and when the liquid does not exist inside the reaction container 11.
It was set to 00 rpm, and the magnetic drive 7 was controlled by turning on / off the control device 6, and it was confirmed whether the stirring member 17 rotated well (smoothly) (20 times or more). As a result, it was confirmed that the stirring member 17 rotated favorably in each case.

Next, regarding the discharge state of the waste liquid,
1c is connected to the syringe pump 1 or 2,
Then, 1.5 ml of distilled water was injected into the container, and then the discharge pump 3 was driven to discharge the distilled water to the waste liquid storage container 4. When this experiment was repeated three times, about 50 μl of the residual liquid was found in the reaction vessel 11. This value is about 4% of the injection volume and is within tolerance. That is, it was confirmed that the residual liquid of this degree was sufficient for automatic cleaning, and it was possible to continuously perform sample measurement.

Further, 1.5 ml of distilled water kept at 25 ° C. in advance was poured into the inside of the reaction vessel 11, and 30 minutes after the start of driving of the Peltier device 32,
The liquid temperature of No. 1 was 25 ° C. ± 0.5 ° C.
That is, it was confirmed that the temperature of the liquid inside the reaction vessel 11 can be kept constant by using the Peltier device 32.

<Second Experimental Example> The methamphetamine (MA) using an anti-methamphetamine monoclonal antibody was measured using the sample mixing apparatus or the fluorescence detection apparatus according to the present invention by a fluorescence enhancement method. FIG. 6 shows the result. FP-52 manufactured by JASCO Corporation as the fluorescence detecting element 13
0 was used. First, after the temperature of the reaction vessel 11 was set to 25 ° C., the antibody solution pump 1 was driven to drive the reaction solution to 1 × 10 ⁻⁷ M (mo).
1 / l) Inject 1.5 ml of the antibody solution into the reaction vessel 11, rotate the stirring member 17 to stir the antibody solution (reagent), and then measure the fluorescence (excitation wavelength: 280 nm, measurement wavelength: 340 nm) ). Next, while the stirring member 17 is being rotated, the sample liquid pump 2 is driven to inject 80 μl of 1 × 10 ⁻⁶ M methamphetamine into the reaction vessel 11,
The fluorescence was measured again (excitation wavelength: 280 nm, measurement wavelength: 340 nm). As a result, it was confirmed that the fluorescence intensity increased by 10% as shown in FIG. The time required for the above measurement was about 60 seconds. Next, the discharge pump 3 is operated to discharge the liquid to be measured into the waste liquid storage container 4.
And the waste liquid (methamphetamine) remaining inside the reaction vessel 11 was washed. Thereafter, the next sample was measured according to the above procedure.

[0020]

As described above, according to the sample mixing apparatus of the present invention and the fluorescence detecting apparatus using the same, the inlet is provided at the top of the reaction vessel and the discharge nozzle is provided at the bottom, and the cleaning liquid is injected from the inlet. Then, by rotating the stirring member to wash the inside of the reaction vessel and discharging the waste liquid from the discharge nozzle, the inside of the reaction vessel is automatically washed, so that
It is possible to measure a plurality of samples continuously. Further, the relative distance between the stirring member and the magnetic drive device for driving the stirring member can be adjusted by the adjusting mechanism, and the stirring member can be surely rotated at a predetermined rotation speed. Further, since the Peltier element is provided, the temperature of the reaction vessel can be kept constant, and the measurement of the sample can be performed without being affected by the surrounding temperature conditions. Further, since the holder is made of aluminum having high thermal conductivity, the time required for temperature adjustment can be shortened. Furthermore, since the surface of the holder is subjected to black alumite treatment, light scattering during fluorescence detection can be reduced, and measurement accuracy can be improved.

[Brief description of the drawings]

FIG. 1 (a) is a front view showing the configuration of a first embodiment of a sample mixing device or a fluorescence detection device of the present invention, and FIG. 1 (b) is a side view thereof.

FIG. 2 is a block diagram illustrating a schematic configuration of a sample mixing device or a fluorescence detection device according to a first embodiment configured to continuously measure a plurality of samples.

FIG. 3 is a perspective view showing a detailed configuration of a reaction vessel in FIGS. 1 and 2;

FIG. 4 (a) is a front view showing the configuration of a second embodiment of the sample mixing device or the fluorescence detection device of the present invention, and FIG. 4 (b) is a side view thereof.

FIG. 5 is a block diagram illustrating a schematic configuration of a sample mixing device or a fluorescence detection device according to a second embodiment configured to continuously measure a plurality of samples.

FIG. 6 is a diagram showing the results of measuring the fluorescence intensity of methamphetamine using the fluorescence detection device according to each example of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Pump for antibody liquid 2 Pump for sample liquid 3 Pump for discharge 4 Waste liquid storage container 5 Sample mixing device or fluorescence detection device 6 Control device 7 Magnetic drive device 10 Holder 11 Reaction container 11b Discharge nozzle 11c Inlet 12 Adjustment mechanism (adjustment screw 13) Fluorescence detection element 17 Stirring member 32 Peltier element 33 Heat radiation fin

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-58-62564 (JP, A) Japanese Utility Model Application Hei 5-3985 (JP, U) (58) Fields investigated (Int. Cl. ⁷ , DB name) G01N 35/00-35/10 B01F 13/08 G01N 21/03 G01N 21/64

Claims (2)

(57) [Claims] 1. An injection port and an air vent hole are provided at an upper part,
A reaction vessel having a rectangular cross section having a discharge nozzle at a lower portion, a stirring member provided inside the reaction vessel and near the bottom thereof, and rotated by a magnetic force from outside the reaction vessel; A magnetic drive device that is provided near the stirring member and generates a magnetic field for rotating the stirring member, and an adjusting mechanism for adjusting a relative distance between the stirring member and the magnetic drive device. And a Peltier element for temperature control in the vicinity of the reaction vessel.
Provided, the reaction vessel, a magnetic drive, an adjustment mechanism and Peltier element
Black anodized surface to hold offspring
A sample measuring device comprising an aluminum holder . 2. It has an inlet and an air vent hole at the top,
A reaction vessel having a rectangular cross section having a discharge nozzle at a lower portion, a stirring member provided inside the reaction vessel and near the bottom thereof, and rotated by a magnetic force from outside the reaction vessel; And a magnetic drive device that is provided near the stirring member and generates a magnetic field for rotating the stirring member; an adjustment mechanism for adjusting a relative distance between the stirring member and the magnetic drive device; provided in the vicinity of the reaction vessel and a fluorescence detection device for detecting the fluorescence of a sample in the reaction vessel, the Peltier device for temperature control in the vicinity of the reaction vessel
Provided, the reaction vessel, a magnetic drive, an adjustment mechanism and Peltier element
Black anodized surface to hold offspring
A fluorescence detection device comprising an aluminum holder .