JPH0712821A - Sample mixing apparatus and fluorescence detection apparatus using the same - Google Patents

Abstract

(57) [Summary] [Purpose] To obtain a sample mixing device or a fluorescence detection device capable of continuous measurement while keeping the temperature of the sample constant and stirring uniformly. [Structure] An inlet 11c is provided at the top of the reaction vessel 11 and a discharge nozzle 11b is provided at the bottom, and the inside of the reaction vessel 11 is automatically washed by injecting a cleaning liquid from the inlet 11c and discharging a waste liquid from the discharge nozzle 11b. To do. Adjustment mechanism 1
The relative distance between the stirring member 17 and the magnetic drive device 7 for driving the stirring member 17 is adjusted by 2 to surely rotate the stirring member 17 at a predetermined rotation speed. A Peltier element 32 is provided to keep the temperature of the reaction vessel 11 constant.

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 such as a fluorescence detecting device used for measuring the characteristics of fluorescent substances in the fields of chemistry, medicine, analytical chemistry and the like.

[0002]

2. Description of the Related Art In the fields of chemistry, medicine, analytical chemistry, and the like, development of fluorescence detection devices and the like has been advanced in order to measure characteristics of fluorescent substances and the like. When measuring multiple samples using a fluorescence detector, etc., as generally called batch processing,
After the measurement of the sample No. 1 is completed, the reaction container, which is the sample measurement unit of the fluorescence detection device, is removed from the holder, and after cleaning, the reaction container is attached again to the holder to prepare for the next sample analysis. 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 container, and a driving mechanism for driving the stirring member reacts. It is provided outside the container. Further, since the fluorescence intensity varies depending on the temperature change in the summer or winter, constant temperature water or the like is circulated from the constant temperature water tank to the reaction container to keep the temperature constant. In addition, since the amount of sample to be measured at one time is as small as several ml and the reaction container is also small accordingly, the stirring device that can be mounted inside the reaction container must be small, which is complicated. It is not possible to use one with a different structure. Therefore, the stirring is composed of a stirring member (magnetic star rubber) provided independently inside the reaction vessel and a magnetic drive device (magnetic stirrer) provided at the bottom of the reaction vessel to rotate the stirring member by magnetic force. The device is being used. The configuration of the conventional sample mixing apparatus as described above is well known or obvious, and therefore the illustration is omitted.

[0003]

When performing measurement using a conventional sample mixing apparatus, the reaction container must be removed from the holder and washed for each measurement of the sample, and continuous measurement of a plurality of samples is required. It was virtually impossible to do.
Also, since the reaction vessel itself is small, it is not possible to use a magnetic drive device with a strong magnetic force (large size), and the distance between the stirring member inside the reaction vessel and the magnetic drive device is less than the predetermined distance due to installation errors or the like. If the length is too long, the strength of the magnetic field acting on the stirring member becomes weak, and there is a possibility that the stirring member may fail to rotate. Further, in order to keep the temperature constant, since the constant temperature water is circulated from the constant temperature water tank to the hollow reaction container holder, the size of the apparatus becomes large, the structure becomes complicated, and maintenance is required.

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

The present invention has been made in order to solve the above problems, and enables continuous measurement of a plurality of samples, does not cause defective stirring, and is easy to maintain and small in size. An object is to provide an apparatus and a fluorescence detection apparatus using the apparatus.

[0006]

The sample mixing apparatus of the present invention comprises a reaction vessel having a rectangular cross section having an inlet and an air vent hole in the upper portion and a discharge nozzle in the lower portion, and the inside of the reaction vessel. And a stirring member that is provided near the bottom of the reaction container and that is rotated by a magnetic force from the outside of the reaction container, and a magnetic field that is provided outside the reaction container and near the stirring member to rotate the stirring member. And a magnetic drive device, and an adjusting mechanism for adjusting the relative distance between the stirring member and the magnetic drive device.

Further, the fluorescence detection device according to the present invention has a reaction container having a rectangular cross section having an injection port and an air vent hole in the upper part and a discharge nozzle in the lower part, and inside and inside the reaction container. A stirring member that is provided in the vicinity of the bottom portion and that is rotated by a magnetic force from the outside of the reaction container, and a magnetic drive that is provided outside the reaction container and in the vicinity of the stirring member and that generates a magnetic field for rotating the stirring member. Device,
An adjustment mechanism for adjusting the relative distance between the stirring member and the magnetic drive device, and a fluorescence detection element provided in the vicinity of the reaction container for detecting the fluorescence of the sample in the reaction container are provided. .

In the above structure, it is preferable to further provide a Peltier element for temperature control near the reaction vessel. Further, it is preferable to use a holder made of aluminum whose surface is black alumite treated in order to hold the reaction container, the magnetic drive device, the adjusting mechanism and the Peltier element.

[0009]

In the sample mixing apparatus of the present invention and the fluorescence detection apparatus using the same configured as described above, an injection port is provided at the top of the reaction container 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 multiple samples can be continuously measured. It is possible. Further, by adjusting the relative distance between the stirring member and the magnetic drive device for driving the stirring member to a regular distance by the adjusting mechanism, the stirring member rotates reliably and at a predetermined rotation speed. Further, since the Peltier device is provided and the temperature of the reaction container is kept constant, the sample can be measured without being affected by the ambient temperature conditions. Furthermore, since the holder is made of aluminum having high thermal conductivity, the time required for temperature adjustment is shortened. Furthermore, since the surface of the holder is black-anodized, 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 a preferred first embodiment thereof. Figure 1 (a)
Is a front view showing the configuration of a sample mixing device or a fluorescence detection device,
(B) is the side view. FIG. 2 is a block diagram showing a schematic configuration of a sample mixing apparatus or a fluorescence detection apparatus configured to continuously measure a plurality of samples. FIG. 3 is a perspective view showing a detailed configuration of the reaction container in FIGS. 1 and 2.

In FIG. 1, a sample mixing apparatus or fluorescence detection apparatus 5 of the present invention comprises a holder 10 made of aluminum, a reaction container 11 fixed to the holder 10, and an adjusting mechanism (adjusting screw) provided in the holder 10. Reaction container 1 through 12
1, a magnetic drive device (magnetic stirrer) 7 provided outside and underneath 1, a stirring member (magnetic star rubber) 17 provided inside the reaction container 11 and near the bottom, and near the reaction container 11. It is composed of a fluorescence detection element (or another detection element) 13 and the like provided in the. 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, for example, a square cell (having a rectangular cross section) of 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. Figure 1
As shown in (b), the magnetic drive device 7 is fixed so that it can be adjusted up and down by an adjusting mechanism (adjusting screw) 12 provided on the bottom of the holder 10. As a result, the relative distance to the stirring member 17 provided inside the reaction container 11 can be made as close as possible to 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 a high thermal conductivity is used as the material of the holder 10 and is used as a fluorescence detection device.

As shown in FIG. 2, for example, two inlets 11c provided in the upper portion of the reaction vessel 11 are connected to syringe-type pumps 1 and 2, respectively, and a discharge nozzle 11b is used for storing waste liquid via a discharge pump 3. It is connected to the container 4. The pump 1 is used, for example, to control the injection amount of the antibody solution which is a reagent, and the pump 2 is used to control the injection amount of the sample liquid. Further, a 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 a preferred second embodiment 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 showing a schematic configuration of a sample mixing device or a fluorescence detection device configured to continuously measure a plurality of samples.
Note that members given the same numbers as those in the first embodiment are substantially the same, so description thereof will be omitted.

In FIGS. 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.
It is m × 23 mm, and is controlled with the power supply current kept constant. The radiation fins 33 are made of aluminum in order to improve the temperature control efficiency.

<First Experimental Example> Using the sample mixing apparatus or the fluorescence detection apparatus according to the first and second examples, the stirring member 17 is rotated, the waste liquid is discharged from the reaction vessel 11, and the second example is used. The temperature control of the liquid such as the sample in the reaction container 11 in Example 1 was measured.

First, regarding the rotation of the stirring member 17, the number of rotations of the stirring member 17 is 8 when the liquid is present inside the reaction vessel 11 and when it is not present.
It was set to 00 rpm, and the magnetic drive device 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 well in any case.

Next, regarding the discharge status of the waste liquid, the inlet 1
1c is connected to the syringe pump 1 or 2 and the reaction container 11
1.5 ml of distilled water was injected into the inside of the container, and then the discharge pump 3 was driven to discharge the distilled water into the waste liquid storage container 4. When this experiment was repeated 3 times, there was about 50 μl of residual liquid in the reaction vessel 11. This value is about 4% of the injected amount and is within the tolerance. That is, it was confirmed that the residual liquid of this level is sufficient for automatic cleaning, and continuous sample measurement is possible.

Further, 1.5 ml of distilled water, which was previously kept at 25 ° C., was injected into the reaction vessel 11 and the reaction vessel 1 30 minutes after the start of driving the Peltier element 32.
When the liquid temperature of 1 was measured, it was 25 ° C. ± 0.5 ° C.
That is, it was confirmed that the temperature of the liquid inside the reaction container 11 can be kept constant by using the Peltier device 32.

<Second Experimental Example> The measurement of methamphetamine (MA) using an anti-methamphetamine monoclonal antibody was carried out by the fluorescence enhancement method using the sample mixing apparatus or the fluorescence detection apparatus according to the present invention. The result is shown in FIG. FP-52 manufactured by JASCO Corporation as the fluorescence detection 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 ^{operate at} 1 × 10 ⁻⁷ M (mo
l / l) 1.5 ml of the antibody solution was injected into the reaction vessel 11, the stirring member 17 was rotated to stir the antibody solution (reagent), and then the fluorescence was measured (excitation wavelength: 280 nm, measurement wavelength: 340 nm). ). Next, with the stirring member 17 rotated, the pump 2 for the sample solution was 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 antibody liquid pump 1 is driven to drive 2.5 ml of the antibody liquid into the reaction container 11.
The waste liquid (methamphetamine) remaining inside the reaction vessel 11 was washed. Then, the following samples were measured according to the above procedure.

[0020]

As described above, according to the sample mixing apparatus of the present invention and the fluorescence detection apparatus using the same, the injection port is provided at the top of the reaction container and the discharge nozzle is provided at the bottom, and the cleaning liquid is injected from the injection port. 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.
It is possible to continuously measure a plurality of samples. Moreover, 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 reliably rotated at a predetermined rotation speed. Further, since the Peltier element is provided, the temperature of the reaction container can be kept constant, and the sample can be measured without being affected by the ambient temperature conditions. Further, since the holder is made of aluminum having high thermal conductivity, the time required for temperature adjustment can be shortened. Further, since the surface of the holder is black-anodized, it is possible to reduce light scattering during fluorescence detection and improve measurement accuracy.

[Brief description of drawings]

1A is a front view showing the configuration of a first embodiment of a sample mixing apparatus or a fluorescence detection apparatus of the present invention, and FIG. 1B is a side view thereof.

FIG. 2 is a block diagram showing a schematic configuration of a sample mixing apparatus or a fluorescence detection apparatus according to a first example configured to continuously measure a plurality of samples.

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

4A is a front view showing the configuration of a second embodiment of the sample mixing apparatus or the fluorescence detection apparatus of the present invention, and FIG. 4B is a side view thereof.

FIG. 5 is a block diagram showing a schematic configuration of a sample mixing apparatus or a fluorescence detection apparatus according to a second example 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]

 1 Pump for Antibody Solution 2 Pump for Sample Solution 3 Pump for Discharge 4 Waste Liquid Storage Container 5 Sample Mixer or Fluorescence Detection Device 6 Control Device 7 Magnetic Drive Device 10 Holder 11 Reaction Vessel 11b Discharge Nozzle 11c Injection Port 12 Adjustment Mechanism (Adjustment Screw ) 13 Fluorescence detecting element 17 Stirring member 32 Peltier element 33 Radiating fin

Claims (6)

[Claims] 1. An injection port and an air vent hole are provided in an upper portion,
A reaction container having a rectangular cross section having a discharge nozzle at the bottom, a stirring member provided inside the reaction container and in the vicinity of the bottom thereof, and rotated by a magnetic force from the outside of the reaction container, and the outside of the reaction container And a magnetic drive device that is provided in the vicinity of the stirring member and that generates a magnetic field for rotating the stirring member, and an adjusting mechanism that adjusts a relative distance between the stirring member and the magnetic driving device. Sample mixing device. 2. The sample mixing apparatus according to claim 1, wherein a Peltier element for temperature control is provided near the reaction vessel. 3. The sample mixing apparatus according to claim 2, wherein a holder made of aluminum whose surface is black-anodized is used to hold the reaction container, the magnetic drive unit, the adjusting mechanism, and the Peltier device. 4. An injection port and an air vent hole are provided at the top,
A reaction container having a rectangular cross section having a discharge nozzle at the bottom, a stirring member provided inside the reaction container and in the vicinity of the bottom thereof, and rotated by a magnetic force from the outside of the reaction container, and the outside of the reaction container A magnetic drive device that is provided in the vicinity of the stirring member and that generates a magnetic field for rotating the stirring member; an adjusting mechanism that adjusts the relative distance between the stirring member and the magnetic driving device; A fluorescence detection device, which is provided in the vicinity of a reaction container and comprises a fluorescence detection element for detecting the fluorescence of the sample in the reaction container. 5. The fluorescence detection device according to claim 4, wherein a Peltier element for temperature control is provided near the reaction container. 6. The fluorescence detection device according to claim 5, wherein a holder made of aluminum whose surface is black-anodized is used to hold the reaction container, the magnetic drive device, the adjusting mechanism, and the Peltier element.