阅读说明：本技术 荧光探针环境参数响应特性的自动检测装置及检测方法 (Automatic detection device and detection method for response characteristics of environmental parameters of fluorescent probe ) 是由 石吉勇 李文亭 邹小波 黄晓玮 李志华 胡雪桃 郭志明 张文 于 2019-08-26 设计创作，主要内容包括：本发明属于分析测试技术领域,具体涉及一种荧光探针环境参数响应特性的自动检测装置及检测方法；本发明所述装置包括荧光光源模块,用于产生光源；荧光信号检测模块,用于检测荧光探针待测液对应的荧光信号；环境参数感知模块,用于感应光线强度、荧光探针待测液温度、荧光探针待测液pH值；环境参数调控模块,用于调控荧光探针待测液环境参数；中央控制器,分别与荧光光源模块、荧光信号检测模块、环境参数感知模块、环境参数调控模块和计算机连接；本发明所述的检测装置可通过环境参数调节模块自动、定向的调节、维持检测体系对应的环境参数,高效、自动获取不同检测环境参数对应的荧光探针响应信号,实现荧光探针环境参数响应特性的高效检测。(The invention belongs to the technical field of analysis and test, and particularly relates to an automatic detection device and a detection method for environmental parameter response characteristics of a fluorescent probe; the device comprises a fluorescent light source module, a light source module and a control module, wherein the fluorescent light source module is used for generating a light source; the fluorescent signal detection module is used for detecting a fluorescent signal corresponding to the to-be-detected liquid of the fluorescent probe; the environment parameter sensing module is used for sensing the light intensity, the temperature of the liquid to be detected by the fluorescent probe and the pH value of the liquid to be detected by the fluorescent probe; the environment parameter regulating and controlling module is used for regulating and controlling the environment parameters of the liquid to be detected of the fluorescent probe; the central controller is respectively connected with the fluorescent light source module, the fluorescent signal detection module, the environmental parameter sensing module, the environmental parameter regulating and controlling module and the computer; the detection device can automatically and directionally adjust and maintain the environmental parameters corresponding to the detection system through the environmental parameter adjusting module, efficiently and automatically acquire the fluorescent probe response signals corresponding to different detection environmental parameters, and realize the efficient detection of the environmental parameter response characteristics of the fluorescent probe.)

1. The automatic detection device of the environmental parameter response characteristic of the fluorescent probe is characterized by comprising:

the fluorescent signal detection module is used for detecting a fluorescent signal of the liquid to be detected of the fluorescent probe;

the environmental parameter sensing module is used for sensing target environmental parameters;

the environment parameter regulating and controlling module is used for regulating and controlling the environment parameters of the liquid to be detected of the fluorescent probe; and

and the central controller is used for controlling the fluorescence signal detection module, the environmental parameter sensing module and the environmental parameter regulation and control module according to instructions.

2. The apparatus for automatic detection of the environmental parameter response characteristic of a fluorescent probe according to claim 1, further comprising a fluorescent light source module for generating a light source;

the device also comprises a computer, and the computer is used for inputting control parameters, storing data and displaying detection results by a user.

3. The apparatus of claim 2, wherein the central controller is communicatively connected to the fluorescent light source module, the fluorescent signal detection module, the environmental parameter sensing module, the environmental parameter control module, and the computer (34), respectively.

4. The apparatus for automatically detecting the environmental parameter response characteristics of a fluorescent probe according to claim 2, wherein the fluorescent light source module comprises a fluorescent light source (1), a light source condenser (2) and a spectroscope (3); light source light (5) emitted by the fluorescent light source (1) passes through the light source condenser (2) and then passes through the spectroscope (3) to be divided into two paths of light, wherein one path of light enters the sample cell (8), and the other path of light enters the environmental parameter sensing module.

5. The apparatus for automatically detecting the environmental parameter response characteristic of a fluorescent probe according to claim 1, wherein the fluorescent signal detection module comprises a fluorescent light condenser (10), a detector probe (11), a detector controller (13); the detector controller (13) is respectively connected with the detector probe (11) and the central controller (4).

6. The apparatus for automatically detecting the environmental parameter response characteristic of a fluorescent probe as claimed in claim 1, wherein the environmental parameter sensing module comprises a light intensity sensor (15), a temperature sensor (16) and a pH sensor (17); light intensity inductor (15), temperature sensor (16), pH value sensor (17) all link to each other with central controller (4), light intensity inductor is used for perception light source light intensity, and temperature sensor is used for the temperature of perception fluorescence probe liquid of waiting to be measured, and pH value sensor is used for the temperature pH value of perception fluorescence probe liquid of waiting to be measured.

7. The apparatus according to claim 1, wherein the environment parameter adjusting and controlling module comprises a light source intensity adjusting sub-module, a pH value adjusting sub-module, a fluorescent probe concentration adjusting sub-module and a temperature adjusting sub-module;

the light source intensity adjusting submodule is used for adjusting and controlling the light intensity of the fluorescent light source (1);

the pH value adjusting submodule is used for adjusting the pH value of the to-be-detected liquid (9) of the fluorescent probe;

the fluorescent probe concentration adjusting submodule is used for adjusting the volume of the fluorescent probe to-be-detected liquid (9) and the probe concentration; and the temperature adjusting submodule is used for adjusting the temperature of the liquid (9) to be detected of the fluorescent probe.

8. The apparatus for automated detection of the environmental parameter response characteristics of fluorescent probes according to claim 1, wherein said environmental parameter regulation module comprises a stirring submodule comprising a magnetic stirrer (29) connected to a central controller (4).

9. The apparatus of claim 2, wherein the fluorescent light source module, the fluorescent signal detection module, the environmental parameter sensing module, and the environmental parameter control module are disposed inside a device housing capable of isolating environmental light.

10. A method for automatically detecting the response characteristic of the environmental parameter of a fluorescent probe is characterized by comprising the following steps:

the automatic detection device for the environmental parameter response characteristics of the fluorescent probe according to any one of claims 1 to 9 is adopted to complete the following steps: the environment parameter sensing module senses the current environment parameter value in real time, the central controller (4) automatically controls the environment parameter regulating and controlling module to regulate the current environment parameter value to the starting point of a response parameter of a preset environment parameter, and the fluorescence signal detecting module detects a fluorescence signal corresponding to the to-be-detected liquid of the fluorescence probe; according to the set adjustment interval and the adjustment times, the central controller (4) automatically controls the environmental parameter adjustment module to sequentially adjust the current environmental parameter value to the preset environmental parameter value, and utilizes the fluorescence signal detection module to sequentially detect the fluorescence signal corresponding to the liquid to be detected of the fluorescence probe under the condition of the preset environmental parameter, namely, the fluorescence signal is used as the response characteristic of the fluorescence probe to the environmental parameter.

11. The method for automatically testing the environmental parameter response characteristics of a fluorescent probe according to claim 10, wherein the process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value comprises: when the environmental parameter is the light source intensity, the current light intensity is compared with the preset light source intensity, if the current light intensity is inconsistent with the preset light source intensity, the central controller (4) controls the light source intensity adjusting submodule to regulate and control the current light intensity, and the light intensity of the fluorescent light source is kept constant until the current light intensity is equal to the preset light source intensity.

12. The method for automatically testing the environmental parameter response characteristics of a fluorescent probe according to claim 10, wherein the process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value comprises: when the environmental parameter is the pH value, comparing the current pH value of the to-be-detected liquid of the fluorescent probe with a preset pH value, if the current pH value is inconsistent with the preset pH value, controlling a pH regulating liquid constant delivery pump controller (22) in a pH value regulating submodule by a central controller (4) to drive a pH regulating liquid constant delivery pump (21) to pump a pH regulating solution (20) into a sample pool (8) through a pH regulating liquid delivery pipe (23), simultaneously driving a magnetic stirrer (30) to uniformly mix the solution in the sample pool (8) by the central controller (4) through a magnetic stirrer (29) of a stirring submodule, comparing the current pH value of the to-be-detected liquid of the fluorescent probe sensed by a pH value sensor (17) in real time with the preset pH value, and stopping the pumping of the pH regulating solution until the current pH value is consistent with the preset pH value.

13. The method for automatically testing the environmental parameter response characteristics of a fluorescent probe according to claim 10, wherein the process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value comprises: when the environmental parameter is the probe concentration, calculating the volume of the probe adjusting solution to be added when the fluorescence probe to be detected liquid reaches the preset probe concentration according to the current volume of the fluorescence probe to be detected liquid, the current probe concentration of the fluorescence probe to be detected liquid and the probe concentration of the probe adjusting solution (25), driving the probe adjusting solution constant delivery pump (26) to pump the probe adjusting solution (25) into the sample pool (8) by the central controller (4) through the probe adjusting solution constant delivery pump controller (27) in the fluorescence probe concentration adjusting submodule, and simultaneously driving the magnetic stirrer (30) to uniformly mix the solution in the sample pool (8) by the central controller (4) through the magnetic stirrer (29) of the stirring submodule.

14. The method for automatically testing the environmental parameter response characteristics of a fluorescent probe according to claim 10, wherein the process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value comprises: when the environmental parameter is temperature, the central controller (4) compares the current temperature of the liquid to be detected of the fluorescent probe with the preset temperature, if the current temperature is not consistent with the preset temperature, the central controller (4) controls the heating sheet (31) to start working through the temperature regulation submodule temperature controller (33), and when the current temperature is consistent with the preset temperature, the current temperature of the liquid to be detected of the fluorescent probe is kept constant; if the current temperature is consistent with the set temperature, the heating sheet (31) and the cooling sheet (32) do not work.

Technical Field

The invention belongs to the technical field of analysis and test, and particularly relates to an automatic detection device and a detection method for environmental parameter response characteristics of a fluorescent probe.

Background

The fluorescent probe has the characteristics of strong tissue penetrability, high efficiency, sensitivity, real-time and convenient detection and the like, and is widely used in the fields of food detection, environmental monitoring, biological imaging and the like. The high sensitivity of the fluorescent probe also means that the fluorescent probe is extremely easily influenced by environmental parameters (temperature, pH value and the like) of a detection system, so that the detection of the environmental parameter response characteristics of the fluorescent probe is very important for the reliability and stability of the detection result of the fluorescent probe.

At present, the fluorescence probe signal is mainly detected by a commercial fluorescence spectrophotometer, and the equipment can accurately and quickly detect the fluorescence signal generated by the fluorescence probe. However, due to the limitations of production cost, difficulty in operation and the like, commercial fluorescent signal detection equipment does not have the functions of sensing environmental parameters of a detection system in real time and automatically regulating and controlling the environmental parameters. The detection of the environmental parameter response characteristics of the fluorescent probe requires obtaining fluorescent signals of the environmental parameters at different levels, and when commercial fluorescent signal detection equipment detects the environmental parameter response characteristics of the fluorescent probe, the environmental parameters of the probe solution can only be adjusted to an expected level before the probe solution is placed in the fluorescent signal detection equipment. The corresponding disadvantages are as follows: (1) the environmental parameter level adjustment of the probe solution is carried out manually, and the efficiency is low; after the probe solution with the environmental parameters adjusted to the specific level is placed into commercial detection equipment, the corresponding environmental parameters are easy to drift, and the existing equipment cannot sense the drift condition of the environmental parameters and cannot make dynamic compensation.

Disclosure of Invention

The invention provides an automatic detection device and a detection method for the environmental parameter response characteristic of a fluorescent probe, aiming at accurately and automatically regulating and controlling the environmental parameter of a fluorescent probe solution and further efficiently detecting the environmental parameter response characteristic of the fluorescent probe.

The automatic detection device for the environmental parameter response characteristics of the fluorescent probe comprises a fluorescent signal detection module, a fluorescence signal detection module and a fluorescence signal detection module, wherein the fluorescent signal detection module is used for detecting a fluorescent signal of a liquid to be detected of the fluorescent probe; the environmental parameter sensing module is used for sensing target environmental parameters;

the environment parameter regulating and controlling module is used for regulating and controlling the environment parameters of the liquid to be detected of the fluorescent probe; and the central controller is used for controlling the fluorescence signal detection module, the environmental parameter sensing module and the environmental parameter regulation and control module according to instructions.

The device also comprises a fluorescent light source module for generating a light source; the device also comprises a computer, and the computer is used for inputting control parameters, storing data and displaying detection results by a user.

The central controller is respectively in communication connection with the fluorescent light source module, the fluorescent signal detection module, the environmental parameter sensing module, the environmental parameter regulating and controlling module and the computer.

The fluorescence light source module consists of a fluorescence light source, a light source condenser and a spectroscope; light of a light source emitted by the fluorescent light source passes through the light source condenser and then the beam splitter and then is split into two paths of light, wherein one path of light enters the sample cell, and the other path of light enters the environmental parameter sensing module; the light source condenser lens is changed into parallel light rays, and the parallel light rays are divided into light intensity sensing light rays and fluorescence exciting light rays after passing through the light splitter lens.

The fluorescence signal detection module comprises a fluorescence light condenser, a detector probe and a detector controller; the detector controller is respectively connected with the detector probe and the central controller. The fluorescence excitation light from the fluorescence light source module irradiates into the liquid to be detected of the fluorescence probe in the sample cell to generate emission fluorescence light, the emission fluorescence light enters the detector probe after passing through the fluorescence light condenser, and the detected fluorescence signal enters the detector controller through the detector data line.

The environment parameter sensing module comprises a light intensity sensor, a temperature sensor and a pH value sensor; wherein the light intensity sensor, the temperature sensor and the pH value sensor are all connected with the central controller; the light intensity sensor is used for sensing the light intensity of the light source, the temperature sensor is used for sensing the temperature of the liquid to be detected of the fluorescent probe, and the pH value sensor is used for sensing the temperature and the pH value of the liquid to be detected of the fluorescent probe.

The environment parameter regulation and control module comprises a light source intensity regulation submodule, a pH value regulation submodule, a fluorescent probe concentration regulation submodule, a stirring submodule and a temperature regulation submodule.

The light source intensity adjusting submodule comprises a light source intensity adjuster connected with the central controller, and the light source intensity adjuster is used for adjusting the current light intensity of the fluorescent light source to the parameter starting point of the light source intensity.

The pH value adjusting sub-module comprises a pH adjusting liquid pool, a pH adjusting solution, a pH adjusting liquid constant delivery pump controller and a pH adjusting liquid delivery pipe. Wherein the pH regulating liquid quantitative pump is positioned in the pH regulating liquid conveying pipe and is connected with the central controller through a pH regulating liquid quantitative pump controller, and the pH regulating liquid conveying pipe is communicated with the sample pool.

The fluorescent probe concentration regulator sub-module comprises a probe regulating solution pool, a probe regulating solution quantitative pump controller and a probe regulating solution conveying pipe; wherein the probe adjusting solution quantitative pump is positioned inside the probe adjusting solution conveying pipe and is connected with the central controller through a probe adjusting solution quantitative pump controller, and the probe adjusting solution conveying pipe is communicated with the sample cell.

The stirring submodule comprises a magnetic stirrer and a magnetic stirrer, the sample cell is arranged on the magnetic stirrer, the magnetic stirrer is connected with the central controller, and the magnetic stirrer is positioned in the sample cell.

The temperature regulation submodule comprises a heating plate, a cooling plate and a temperature controller; wherein the heating pieces are symmetrically arranged on two sides of the sample cell, the cooling pieces are symmetrically arranged on two sides of the sample cell, and the heating pieces and the cooling pieces are connected with the central controller through the temperature controller.

The fluorescent light source module, the fluorescent signal detection module, the environmental parameter sensing module and the environmental parameter regulation and control module are all arranged in the device shell capable of isolating environmental light.

The invention also provides an automatic detection method of the response characteristic of the environmental parameter of the fluorescent probe, which comprises the following steps:

the automatic detection device for the environmental parameter response characteristics of the fluorescent probe is adopted to complete the following steps:

the environment parameter sensing module senses the current environment parameter value in real time, the central controller automatically controls the environment parameter regulating and controlling module to regulate the current environment parameter value to the starting point of a response parameter of a preset environment parameter, and the fluorescence signal detecting module detects a fluorescence signal corresponding to the to-be-detected liquid of the fluorescence probe;

according to the set adjustment interval and the adjustment times, the central controller automatically controls the environmental parameter adjustment module to sequentially adjust the current environmental parameter value to the preset environmental parameter value, and the fluorescence signal detection module is used for sequentially detecting the fluorescence signal corresponding to the liquid to be detected of the fluorescence probe under the condition of the preset environmental parameter, namely the fluorescence signal is used as the response characteristic of the fluorescence probe to the environmental parameter.

The process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value is as follows: when the environmental parameter is the light source intensity, the current light intensity is compared with the preset light source intensity, if the current light intensity is inconsistent with the preset light source intensity, the central controller controls the light source intensity adjusting submodule to regulate and control the current light intensity, and the light intensity of the fluorescent light source is kept constant until the current light intensity is equal to the preset light source intensity.

The process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value is as follows: and when the environmental parameter is the pH value, comparing the current pH value of the fluorescent probe to-be-detected liquid with a preset pH value, if the current pH value is inconsistent with the preset pH value, controlling a pH regulating liquid constant pump controller in a pH value regulating submodule by a central controller to drive a pH regulating liquid constant pump to pump the pH regulating solution into a sample pool through a pH regulating liquid conveying pipe, simultaneously driving a magnetic stirrer to uniformly mix the solution in the sample pool through a magnetic stirrer of a stirring submodule by the central controller, comparing the current pH value of the fluorescent probe to-be-detected liquid sensed by a pH value sensor in real time with the preset pH value, and stopping pumping the pH regulating solution until the current pH value is consistent with the preset pH value.

The process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value is as follows: when the environmental parameter is the probe concentration, calculating the volume of the probe adjusting solution to be added when the fluorescent probe to be detected liquid reaches the preset probe concentration according to the current volume of the fluorescent probe to be detected liquid, the current probe concentration of the fluorescent probe to be detected liquid and the probe concentration of the probe adjusting solution, driving the probe adjusting solution constant delivery pump to pump the probe adjusting solution into the sample pool by the central controller through the probe adjusting solution constant delivery pump controller in the fluorescent probe concentration adjusting submodule, and simultaneously driving the magnetic stirrer to uniformly mix the solution in the sample pool by the central controller through the magnetic stirrer of the stirring submodule.

The process of sequentially adjusting the current environmental parameter value to the preset environmental parameter value is as follows: when the environmental parameter is temperature, the central controller compares the current temperature of the liquid to be detected of the fluorescent probe with a preset temperature, if the current temperature is inconsistent with the preset temperature, the central controller controls the heating sheet to start working through the temperature regulation submodule temperature controller, and when the current temperature is consistent with the preset temperature, the current temperature of the liquid to be detected of the fluorescent probe is kept constant; and if the current temperature is consistent with the set temperature, the heating sheet and the cooling sheet do not work.

The invention has the beneficial effects that:

the detection device can sense the environmental parameters corresponding to the detection system in real time by using the environmental parameter sensing module, and automatically and directionally adjust and maintain the environmental parameters corresponding to the detection system by using the environmental parameter adjusting module, so that the response signals of the fluorescent probes corresponding to different detection environmental parameters are efficiently and automatically obtained, and the efficient detection of the response characteristics of the environmental parameters of the fluorescent probes is realized. Has the advantages that:

(1) the invention can realize automatic and directional adjustment of the probe solution to different environmental parameter levels, and the device has high automation degree;

(2) the invention can monitor the change condition of the parameter level of the link in real time in the process of measuring the probe fluorescent signal, and make dynamic compensation, so as to ensure that the actual environmental parameter of the probe solution is highly consistent with the set parameter value;

(3) the invention comprises light source intensity, temperature and pH concentration regulation and control modules, each module can work independently and cooperatively, and can comprehensively represent the environmental parameter response characteristics of the fluorescent probe;

drawings

FIG. 1 is a schematic structural diagram of an automatic detection apparatus for environmental parameter response characteristics of a fluorescent probe according to a preferred embodiment of the present invention;

in the figure: 1-fluorescence light source, 2-light source condenser, 3-spectroscope, 4-central controller, 5-light source light, 6-light intensity induction light, 7-fluorescence excitation light, 8-sample cell, 9-fluorescence probe solution to be tested, 10-fluorescence light condenser, 11-detector probe, 12-detector data line, 13-detector controller, 14-emission fluorescence light, 15-light intensity sensor, 16-temperature sensor, 17-pH value sensor, 18-light source regulator, 19-pH regulating solution cell, 20-pH regulating solution, 21-pH regulating solution quantitative pump, 22-pH regulating solution quantitative pump controller, 23-pH regulating solution delivery pipe, 24-probe regulating solution cell, 24-fluorescence probe solution quantitative pump, and the like, 25-probe adjusting solution, 26-probe adjusting solution quantitative pump, 27-probe adjusting solution quantitative pump controller, 28-probe adjusting solution conveying pipe, 29-magnetic stirrer, 30-magnetic stirrer, 31-heating sheet, 32-cooling sheet, 33-temperature controller and 34-computer.

Detailed Description

In order to make those skilled in the art better understand the technical solution of the present invention, the following description is made in conjunction with the specific embodiments and the accompanying drawings so that those skilled in the art can better understand the technical solution of the present invention.