JP2023059170A - fluorescence detector - Google Patents

Abstract

To provide a fluorescence detector capable of shortening a warm-up time and an automatic analyzing device with the fluorescence detector.SOLUTION: There is used a fluorescence detector that comprises exciting light irradiation means which irradiates a sample with exciting light and fluorescence detecting means which detects fluorescence emitted from the sample, wherein the exciting light irradiation means is controlled by a light source control part to be turned on and off, the fluorescence detecting means acquires respective detection signals of exciting light emission and exciting light extinction, and a signal processing part takes the difference between the exciting light emission and exciting light extinction as a fluorescence detection signal.SELECTED DRAWING: Figure 1

Description

TECHNICAL FIELD The present invention relates to a fluorescence detector provided in an automatic analyzer or the like.

Some clinical examination apparatuses are equipped with a temperature control function and a fluorescence detector in order to accurately measure biochemical reactions. For example, in the nucleic acid amplification method (Patent Document 1), a reagent using a nucleic acid probe decorated with a fluorescent dye and a sample are subjected to a nucleic acid amplification reaction at a specific temperature. can measure nucleic acid amplification of pathogens in real time.

In this case, since the fluorescence from the reagent is weak, the light receiving circuit of the fluorescence detector has a large circuit gain and is easily affected by heat. In order to solve this problem, a technique has been disclosed in which the temperature of the fluorescence detector is adjusted together with the sample to make it less susceptible to heat (Patent Document 2). Since the fluorescence detector is located inside the device, it is less susceptible to sudden changes in environmental temperature. There is a problem that it is difficult to shorten the time from turning on the power until measurement is possible, and that it also leads to an increase in size and cost of the apparatus.

In recent years, portable POCT (Point of Care Testing) testing devices have become popular for simple and accurate measurements in clinical testing. For portable and simple measurement, the device is required to be smaller, and the warm-up time is also required to be shorter. there is

If the volume of the reaction reagent is reduced in order to reduce the size of the device and the cost of the reagent, the fluorescence that can be detected by the device becomes weaker and the effect of heat on the fluorescence detector becomes greater. In order to improve usability, an all-in-one cartridge that integrates the reaction reagents used in the test with other consumables can be used. will have an even greater impact.

By downsizing the device, even if the fluorescence detector does not have a temperature control function, the temperature control function of the reagent is arranged closer to the fluorescence detector. Since the heat from the temperature control function of this reagent is transmitted to the fluorescence detector, the output of the fluorescence detector drifts for a long time until the temperature around the detector reaches equilibrium, making it impossible to shorten the warm-up time. The problem still remained.

JP 2015-031682 A JP 2008-256530 A

SUMMARY OF THE INVENTION An object of the present invention is to solve the aforementioned problems, and to provide a fluorescence detector capable of shortening the warm-up time and an automatic analyzer equipped with the fluorescence detector.

In order to solve the above problems, the present inventors have reached the present invention as a result of extensive studies.
That is, the present invention
A fluorescence detector comprising excitation light irradiation means for irradiating a sample with excitation light and fluorescence detection means for detecting fluorescence emitted from the sample,
The excitation light irradiation means is controlled by the light source control unit to turn on and off,
The fluorescence detection means acquires each detection signal of excitation light ON and excitation light OFF,
The signal processing unit is a fluorescence detector that uses the difference between excitation light on and excitation light off as a fluorescence detection signal.

The detector signal when the excitation light is on contains both the fluorescence of the reagent and the thermal drift of the detector circuit, and the detector signal when the excitation light is off is only the thermal drift of the circuit. By blinking the excitation light, obtaining the detector signal when the light is on and when it is off, and taking the difference between the detector signals when the light is on and when the light is off, it is possible to remove the thermal drift of the detector circuit. As a result, the signal of the fluorescence detector can reach equilibrium in a short time, and the warm-up time can be shortened.

In the case of multiple excitations/fluorescence, for example, 3 excitations and 3 fluorescences, the 3 types of excitation light are turned on sequentially (for example, in the order of 450 nm → 500 nm → 590 nm), and then all lights are turned off. , and the difference can be taken.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to shorten the warm-up time in an automatic analyzer equipped with a fluorescence detector.

1 is a configuration diagram of a fluorescence detector of the present invention; FIG. FIG. 2 is a graph showing temporal changes in fluorescence detector signal of 450 nm excitation 495 nm fluorescence and incubator temperature in Example 1 and Comparative Example 1. FIG. FIG. 2 is a graph showing temporal changes in fluorescence detector signal of 500 nm excitation 545 nm fluorescence and incubator temperature in Example 1 and Comparative Example 1. FIG. FIG. 2 is a diagram showing temporal changes in the fluorescence detector signal (excitation at 590 nm and fluorescence at 645 nm) and incubator temperature in Example 1 and Comparative Example 1; FIG. 4 is an external view of a measurement unit provided with fluorescence detectors for 3 excitations and 3 fluorescences.

EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

In this example, using the fluorescence detector of the present invention shown in FIG. 1, it was verified whether the drift of the fluorescence detector immediately after power-on could be reduced. A transparent and colorless polypropylene container was used as the sample container 10 . As the reaction reagent 12, a solvent in which a diffusion probe modified with a fluorescent dye and a specimen are mixed is used, but since the fluorescence intensity changes with the reaction, the solvent was selected to correspond to the fluorescence before the reaction. As the excitation light irradiation unit 14 and the fluorescence detection unit 15, three sets of excitation light of 450 nm and fluorescence of 495 nm, excitation light of 500 nm and fluorescence of 545 nm, and excitation light of 590 nm and fluorescence of 645 nm were used (see FIG. 5). The control temperature of the temperature control block 16 was set at 46°C.

(Comparative example 1)
After the power was turned on, the temperature control block 16 reached the control temperature of 46° C. about 1 minute after the start of heating. In the fluorescence detection unit 15, the excitation light detection signal continued to fluctuate for about 30 minutes after the temperature control block 16 reached the control temperature. The results are shown in FIGS. 2, 3 and 4. FIG. As a result, the warm-up time from turning on the power to starting the measurement took more than 30 minutes.

(Example 1)
After the power was turned on, the temperature control block 16 reached the control temperature of 46° C. about 1 minute after the start of heating. The three excitation light irradiation units 14 were turned on in order (2.5 sec×3, turned off for 2.5 sec), and the corresponding fluorescence intensity of the fluorescence detection unit 15 was measured. Fluctuations in the detection signal obtained by the fluorescence detection unit 15 became constant in about 3 minutes after the temperature control block 16 reached the control temperature by subtracting the excitation light ON and excitation light OFF detection signals. The results are shown in FIGS. 2, 3 and 4. FIG. It takes about 5 minutes from turning on the power to starting measurement, which is a short warm-up time required for a POCT inspection device.

10: sample container 12: reaction reagent 14: excitation light irradiation unit 14a: excitation light source 14b: optical filter (excitation light side)
14c: Effective diameter of excitation light 15: Fluorescence detector 15a: Light receiving element 15b: Optical filter (fluorescence side)
15c: fluorescent effective diameter 16: temperature control block

Claims (4)

A fluorescence detector comprising excitation light irradiation means for irradiating a sample with excitation light and fluorescence detection means for detecting fluorescence emitted from the sample,
The excitation light irradiation means is controlled by the light source control unit to turn on and off,
The fluorescence detection means acquires each detection signal of excitation light ON and excitation light OFF,
The signal processor is a fluorescence detector that uses the difference between excitation light on and excitation light off as a fluorescence detection signal. 2. A fluorescence detector according to claim 1, comprising a plurality of excitation light irradiation means and fluorescence detection means. 3. The fluorescence detector according to claim 1, comprising three excitation light irradiation means and three fluorescence detection means. An automatic analyzer comprising the fluorescence detector according to any one of claims 1 to 3.

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