CN211785130U

CN211785130U - Biological heavy metal pollution detection device

Info

Publication number: CN211785130U
Application number: CN201922336753.0U
Authority: CN
Inventors: 于娜; 杨希国; 考志强; 杨英姿; 张艳艳; 韦石; 李丽
Original assignee: Liaoning General Fair Testing Co ltd
Current assignee: Liaoning General Fair Testing Co ltd
Priority date: 2019-12-24
Filing date: 2019-12-24
Publication date: 2020-10-27
Anticipated expiration: 2029-12-24

Abstract

The utility model relates to a biotype heavy metal pollution detection device, including detecting element, sample/thallus incubation unit, detection preparation unit and data processing and control module. The detection unit comprises a high-transmittance glass detection vessel, a light emitter and a fluorescence detector. The sample/thallus co-incubation unit comprises a constant temperature tank, a conical culture tube is arranged in the constant temperature tank, and the bottom end of the conical culture tube is connected with a motor; the constant temperature bath is equipped with the upper cover, and it corresponds each toper culture tube and is equipped with the bellying, and each bellying corresponds each toper culture tube and is equipped with three intubate to supply to connect the heavy suspension pipe of culture medium storage tank, buffer solution storage tank and detection preparation unit respectively. The detection preparation unit comprises a heavy suspension pipe and a constant volume pipe, wherein the bottom of the heavy suspension pipe is provided with an oscillating mechanism, and the constant volume pipe is connected with a buffer storage tank and a high-light-transmittance glass detection dish. And (3) diluting the bacterial liquid in a constant volume tube according to a specific multiple, and then sending the diluted bacterial liquid into a high-light-transmittance glass detection dish for fluorescence detection. The data processing and control module is used for processing the data sent by the fluorescence detector.

Description

Biological heavy metal pollution detection device

Technical Field

The utility model belongs to the technical field of heavy metal pollution detection device technique and specifically relates to a biological type heavy metal pollution detection device.

Background

With the development of the industry, a lot of heavy metal ions enter surface water and soil. Especially lead pollution, is toxic to all life. Lead poisoning of human body can cause mental retardation, and has liver and kidney toxicity. Therefore, it is important to detect lead ions in water sources or soil. At present, the mature detection technology for detecting the heavy metal is chemical detection and physical detection, the emerging detection technology is biological detection, and the three methods have advantages. Chemical detection methods typically use other reagents that are themselves contaminating. The physical detection generally needs expensive instruments, such as an inductively coupled plasma emission spectrometer and an inductively coupled plasma mass spectrometer, and has the advantages of high detection sensitivity and specificity, the defects of high equipment cost, high maintenance cost after damage and complex operation, and the most important is that a chemical method and a physical method can only detect the objective concentration of certain heavy metal ions and cannot directly reflect the utilization rate, toxicity and influence of polluting ions on biological organisms. The biological detection is that some microbial strains (such as escherichia coli) which have specific reaction on certain metal ions react with heavy metal ions, so that the microbial strains generate fluorescence change and can generate fluorescence with a specific color under excitation light (the heavy metal ions induce the fluorescent protein expression of certain color of the strains), and the concentration of the heavy metal ions and the toxicity or influence on organisms are detected by detecting the change condition of the fluorescence.

The existing device for biological detection method only comprises a light emitter, a fluorescence detector and a data processing module, the structure is simple, but detection personnel usually need to take a sample (water sample containing heavy metal ions) collected in the field back to a laboratory, and then co-incubate with a specific microbial thallus for 2-4h in a specific culture medium and a certain temperature environment, and in the process, the heavy metal ions induce the specific fluorescent protein of the thallus to express; and then carrying out centrifugal separation on the thalli, carrying out fluorescence detection on the separated thalli, and calculating the heavy metal pollution condition in the water sample. Because of being limited by the device, the current biological detection method needs a plurality of devices to be matched for use, and the detection can not be completed on the spot at the acquisition place.

SUMMERY OF THE UTILITY MODEL

Technical problem to be solved

In order to solve the above-mentioned problem of prior art, the utility model provides a biotype heavy metal pollution detection device not only has the fluorescence detection function, but also is equipped with the unit of incubating altogether of sample and microbial thalli, detects the preparation unit, makes the measurement personnel can gather the ground at the sample, only with the help of the detection of the heavy metal pollution condition in the complete sample of an equipment.

(II) technical scheme

In order to achieve the above object, the utility model discloses a main technical scheme include:

a biotype heavy metal pollution detection device, comprising:

the detection unit comprises a high-transmittance glass detection vessel, a light emitter and a fluorescence detector;

the sample/thallus co-incubation unit comprises a constant temperature tank, wherein one end of the constant temperature tank is provided with a water inlet, and the other end of the constant temperature tank is provided with a water outlet; a conical culture tube is arranged in the thermostatic bath, an opening is formed in the upper end of the conical culture tube, the bottom end of the conical culture tube is arranged in the middle of the bearing and is connected with a motor at the bottom of the thermostatic bath; the conical culture tubes are provided with an upper cover, the lower bottom surfaces of the upper covers are provided with convex parts corresponding to the conical culture tubes, and the convex parts correspondingly extend into the openings of the conical culture tubes; the upper cover is provided with at least three insertion tubes corresponding to the conical culture tubes, the insertion tubes penetrate through the convex parts, one ends of the insertion tubes are inserted into the conical culture tubes, and the other ends of the insertion tubes are positioned outside the upper cover and are respectively connected with a culture medium storage tank, a buffer solution storage tank and a heavy suspension tube of the detection preparation unit through a pump and a pipeline;

the detection preparation unit comprises a constant volume pipe and a heavy suspension pipe, the heavy suspension pipe is connected with the constant volume pipe through a pump and a pipeline, and an oscillating mechanism is arranged at the bottom of the heavy suspension pipe; the constant volume tube is connected with the buffer solution storage tank and the high-light-transmittance glass detection vessel through a pump and a pipeline, the buffer solution is extracted from the buffer solution storage tank to carry out constant volume dilution on the thalli, and the bacterium solution with the constant volume dilution is sent into the high-light-transmittance glass detection vessel for carrying out fluorescence detection;

and the data processing and control module is in communication connection with the light emitter, the fluorescence detector, the motor and the pump of the detection unit so as to control the starting, stopping and operating states of the light emitter, the fluorescence detector, the motor and the pump and receive and process data sent by the fluorescence detector.

According to the present invention, one of the three insertion tubes is a central insertion tube, which is connected to a heavy suspension tube or a waste liquid tank through a pump and a pipeline, and the bottom end of the central insertion tube is lower than the other insertion tubes and is opposite to the bottom center of the conical cultivation tube.

According to a preferred embodiment of the present invention, a temperature monitor and a heating rod are disposed in the thermostatic bath, and a heat insulation layer is disposed outside the thermostatic bath; the temperature monitor and the heating rod are in communication connection with the data processing and control module.

According to the utility model discloses a preferred embodiment, wherein, be equipped with round C type recess on the lateral wall all around of the bellying of upper cover, be equipped with the ball in the C type recess, the ball with the internal face contact of toper culture tube.

According to a preferred embodiment of the present invention, the thermostatic bath is provided with a perforated plate, the perforated plate is made of a heat insulating material, the perforated plate is provided with a plurality of through holes, and the conical cultivation tubes are installed in the through holes and detachably connected to the bearings at the bottom of the thermostatic bath; the opening of the conical culture tube is higher than the upper part of the pore plate.

According to the utility model discloses a preferred embodiment, wherein, the detecting element still includes a confession the socket that high light transmittance glass detected ware installation, the socket is equipped with a plurality of slot, two sides that the slot is relative are the window, can supply light emitter to high light transmittance glass to detect ware emission light, and by fluorescence detector detects at the another side.

According to a preferred embodiment of the present invention, the light emitter is controlled and switched by the data processing and control module to emit light with two or more wavelengths; at least one ultraviolet light; the detection unit further comprises an absorbance detector for detecting the optical density of the bacterial liquid in the high-transmittance glass detection dish.

According to a preferred embodiment of the present invention, the apparatus further comprises a housing, the interior of the housing is divided into two layers, the upper layer is provided with the sample/thallus co-incubation unit and the detection preparation unit, and the lower layer is provided with the detection unit; the shell is also provided with a touch display screen.

(III) advantageous effects

The utility model has the advantages that:

the biological heavy metal pollution detection device of the utility model comprises a detection unit, a sample/thallus co-incubation unit and a detection preparation unit, the sample/thallus co-incubation unit can provide specific thallus to be co-cultured with a tested sample for 3-4h at a proper temperature (35-37 ℃), then automatically centrifuging and standing in the conical culture tube, extracting supernatant, then resuspending and cleaning the thalli at the bottom of the conical culture tube with buffer solution, pumping the resuspended cleaning solution back to the resuspension tube, after the resuspension tube is uniformly oscillated, a certain amount of resuspension liquid is extracted to a quantitative dilution tube, the bacteria are diluted by a certain multiple by a constant volume by the buffer liquid extracted from the buffer liquid storage tank, the preparation work of detection is completed, and the bacteria can be sent into a high-light-transmittance glass detection dish for fluorescence detection or absorbance analysis. Therefore, with the help of the utility model discloses a biotype heavy metal pollution detection device, measurement personnel only need with the help of an equipment, can accomplish the detection of the heavy metal pollution condition in the sample on the spot in sample collection ground.

The amount of the bacteria liquid and the amount of the buffer liquid are extracted in the processes of solution fixing and dilution and are finished by controlling corresponding pumps through the data processing and control module, and errors caused by manual operation can be reduced.

In addition, the number of the conical culture tubes is multiple, and multiple parallel experiments of the same sample can be performed; the number of the high-light-transmittance glass detection dishes is also multiple, and the bacterial liquid from the same conical culture tube can be subjected to multiple parallel detection.

Drawings

FIG. 1 is a schematic structural view of the biological heavy metal contamination detection device of the present invention.

Detailed Description

For a better understanding of the present invention, reference will now be made in detail to the present invention, examples of which are illustrated in the accompanying drawings.

As shown in fig. 1, the biological heavy metal contamination detection apparatus 100 includes a detection unit 10, a sample/cell co-incubation unit 20, a detection preparation unit 30, and a data processing and control module 40.

The detection unit 10 includes a high transmittance glass detection dish 11, a light emitter 12, and a fluorescence detector 13. The high-light-transmittance glass detection dish 11 is a glass utensil with the capacity of 0.5-1ml, and the bacteria liquid to be detected is filled in the glass utensil. High light transmittance glass detects ware 11 and installs in a socket 111, and socket 111 is equipped with a plurality of slot, and two sides that the slot is relative are the window, and the window of one side provides light emitter 12 and detects the ware 11 transmission light to high light transmittance glass, and the window of opposite side then provides fluorescence detector 13 and carries out fluorescence intensity's detection. Taking an escherichia coli for detecting lead ions as an example, the light emitter 12 needs to emit light of 550nm, and the escherichia coli incubated with the lead ions for 3-4 hours has a large amount of red fluorescent protein expressed, emits red fluorescence under excitation light of 550nm, detects the fluorescence intensity by the fluorescence detector 13, and calculates the concentration (bioavailable concentration) of the lead ions in the sample by taking a pre-prepared standard curve as a reference. The absorbance is also a parameter for evaluating the cells, and the absorbance of the test sample solution may be measured at the same time. Thus, preferably, the light emitter 12 can be controlled by the data processing and control module 40 to switch to emit light of more than two wavelengths; at least one ultraviolet light is included, and the other light is light with the wavelength matched with the specific fluorescent protein of the thallus.

The sample/cell co-incubation unit 20 includes a constant temperature bath 21. The thermostatic bath 21 has a water inlet 211 at one end and a water outlet 212 at the other end. The water inlet 211 may be connected to tap water. A temperature monitor 213 and a heating rod 214 are arranged in the thermostatic bath 21, and an insulating layer is arranged outside the thermostatic bath. The temperature monitor 213 and the heating rod 214 are connected to the data processing and control module 40 for controlling the temperature of the water in the thermostatic bath 21 to 35-37 ℃ by programming. Three conical culture tubes 22 are arranged in the thermostatic bath 21, the upper end of each conical culture tube 22 is provided with an opening, and the bottom end of each conical culture tube 22 is arranged in the middle of a bearing 23 and is coupled with a motor 24 at the bottom of the thermostatic bath 21. The thermostatic bath 21 is provided with a pore plate 25, the pore plate 25 is made of heat insulating materials and is provided with 3 through holes, and a conical culture tube 22 is arranged in each through hole. The bottom end of the conical culture tube 22 is provided with a long mounting shaft which is detachably inserted in the middle of a bearing 23 at the bottom of the thermostatic bath 20 and is fixedly connected with a motor 24 in the rotating direction. The opening of the conical culture tube 22 is raised above the well plate 25. The volume of the conical culture tube 22 is 20-30ml, but the culture medium is preferably contained in 1/2-1/3 of the volume. The thermostatic bath 21 is provided with an upper cover 26, one end of the upper cover 26 is rotatably connected to one end of the thermostatic bath 21, the lower bottom surface of the upper cover 26 is provided with a convex part 261 corresponding to the opening of each conical cultivation tube 22, and each convex part 261 corresponds to the opening extending into the conical cultivation tube 22. The upper lid 26 is provided with three insertion tubes corresponding to each conical culture tube 22, the three insertion tubes penetrate through the protruding part 261, one end of each insertion tube is inserted into the conical culture tube 22, and the other end of each insertion tube is located outside the upper lid 26, so as to be respectively connected with the culture medium reservoir 27, the buffer solution reservoir 28 and the heavy suspension tube 31 of the detection preparation unit 30 through the pumps P1 and P2 and the pipelines. One of the three cannulas is a central cannula, which is connected with the resuspension tube 31 or the waste liquid tank 29 through a pump P1, a pipeline, an electromagnetic valve C1 (the other conical culture tubes 22 are C2/C3) and an electromagnetic valve E, wherein the tube opening end at the bottom of the central cannula is lower than the other two cannulas and is opposite to the central position at the bottom of the conical culture tube 22.

The motor 24 can drive the conical culture tube 22 to perform centrifugal rotation, so as to separate thalli at the bottom of the conical culture tube 22 by a separation method. In order to ensure the smooth and stable rotation of the conical cultivation tube 22, a circle of C-shaped grooves are formed on the peripheral side walls of the protrusions 261 of the upper cover 26, the balls 262 are arranged in the C-shaped grooves, and the balls 262 are in contact with the inner wall surface of the conical cultivation tube 22, so that the rotation process of the conical cultivation tube 22 is stabilized.

The detection preparation unit 30 includes a resuspension tube 31 and a constant volume tube 32. The resuspension tube 31 is connected to each conical culture tube 22 by an electromagnetic valve E, a pump P1, an electromagnetic valve C1(C2 or C3) and a central cannula, meanwhile, the resuspension tube 31 is also connected to the constant volume tube 32 by a pump P3 and a pipeline, the bottom of the resuspension tube 31 is provided with an oscillating mechanism 311, and the oscillating mechanism 311 can uniformly mix the thalli in the resuspension tube 31. The constant volume tube 32 is also connected with the buffer solution storage tank 28 through a pipeline, a pump P3 and an electromagnetic valve F, D, and the bottom of the constant volume tube 32 is connected with the high-transmittance glass detection dish 11 through a pump P4 and a pipeline.

As shown in FIG. 1, one of the cannulae was connected to the medium reservoir 27 via a line, solenoid valve A1 (A1, A2, A3 for different conical flasks 22), A0, pump P2, solenoid valve H, and the other cannula was connected to the buffer reservoir 28 via a line, solenoid valve B1 (B1, B2, B3 for different conical flasks 22), B0, pump P2, solenoid valve D. The central cannula is connected to waste solution tank 29 by a line, solenoid valve C1 (C1, C2, C3 for the different conical culture tubes 22), pumps P1, C0, or to resuspension tube 31 by a line, solenoid valve C1, pumps P1, C0. The heavy suspension pipe 31 is connected to the constant volume pipe 32 through a pipeline, a solenoid valve E and a pump P3. Meanwhile, the buffer storage tank 28 is connected to the constant volume tube 32 through a pipeline, an electromagnetic valve D, an electromagnetic valve F and a pump P3, so that a large amount of buffer is added to the constant volume tube 32, and the bacterial liquid is diluted by multiple times so as to perform fluorescence detection. The residual bacteria liquid in the resuspension tube 31 and the constant volume tube 32 enters the waste liquid tank 29 through a pump P3 and an electromagnetic valve C0. It should be noted that the arrangement of the pipeline and the electromagnetic valve is not necessary to be implemented in the manner of fig. 1, and any manner capable of realizing the functions can be implemented as long as the culture medium can be automatically added to the conical culture tube, the centrifuged supernatant can be automatically removed, the bacteria can be automatically washed and collected by the buffer solution, and the bacteria can be automatically oscillated, resuspended uniformly, quantified and diluted in fixed times.

And the data processing and control module 40 is in communication connection with the light emitter 12, the fluorescence detector 13 of the detection unit 10, the motor 24 at the bottom of the thermostatic bath 21, the pumps P1, P2, P3, P4 and the electromagnetic valves, so as to control the starting, stopping and operating states (including the positive and negative rotation of the motor and the positive and negative rotation of the pump) of the light emitter 12, the fluorescence detector 13, the motor 24, the pumps P1, P2, P3, P4 and the like, control the conduction conditions of the electromagnetic valves, and receive and process data sent by the fluorescence detector 13.

Preferably, the utility model discloses a biotype heavy metal pollution detection device 100, this shell is inside to be divided into two-layerly, and the upper strata is equipped with sample/thallus and incubates unit 20 and detection preparation unit 30 altogether, and the lower floor is equipped with detecting element 10, and data processing and control module, and the shell surface can set up the touch-control display screen. The upper lid 26 of the upper sample/cell co-incubation unit 20 can be manually opened by the inspector from the outside.

The utility model discloses a detection principle and process as follows:

after obtaining the test sample and performing the preliminary treatment, the upper lid 26 is opened, an equal amount of sample is injected into each of the three conical culture tubes 22, an equal amount of microbial cells prepared in advance is added to each of the three conical culture tubes 22, and the upper lid 26 is closed. Starting the culture medium adding program, the culture medium prepared in advance in the culture medium pot is fed into the three conical culture tubes 22 by the pump P2, the electromagnetic valves A0, A1, A2 and A3, and the total amount of the culture solution is 1/3 of the volume of the conical culture tubes 22. Starting a heating program of the constant temperature groove 21 to heat the water in the constant temperature groove 21 to 35-37 ℃, and constantly maintaining the temperature for 3-4 h. The temperature monitor 213 monitors the temperature of the water in the thermostatic bath 21 in real time and sends a monitoring signal to the data processing and control module 40, and the data processing and control module 40 determines whether to activate the heating rod 214. During the culture process, the motor 24 intermittently drives the conical culture tube 22 to rotate forward and backward to realize slight stirring. When the culture reaches 4, the motor 24 drives the conical culture tube 22 to rotate rapidly, so that the thalli are centrifuged and deposited at the center of the conical culture tube 22. When the conical culture tube 22 is completely stationary, the clear liquid is slowly drawn into the waste liquid tank 29 by the pump P1 and the center cannula (the solenoid valve C0 is turned on), and when the clear liquid is not drawn, the drawing is stopped. The buffer (buffer dedicated for detection) in the buffer storage tank 28 is fed into the conical culture tube 22 by the pump P2 through the electromagnetic valves B0, B1, B2 and B3, and the bacteria at the bottom of the conical culture tube 22 are washed by a large water pressure to be stirred, and during the washing process, the pump P1 is started at the same time, and the bacteria liquid is sucked out by the central insertion tube (the electromagnetic valves C1\ C2\ C3\ E are conducted) and fed into the heavy suspension tube 31. In the resuspension tube 31, after sufficient shaking, a uniform resuspension solution is formed. Then, a pump P3 is used for quantitatively drawing 50-100 microliters to the constant volume tube 32, and a pump P3 is used for adding the buffer solution in the buffer solution storage tank 28 to the constant volume tube 32 to dilute the 50-100 microliters of the heavy suspension in the constant volume tube 32 by 20-40 times (the cell concentration is overlarge without dilution, and the detection difficulty is large). Thus, the bacteria liquid meeting the detection requirement is prepared and can be sent into the high-transmittance glass detection dish 11 for detection. The light emitter 12 emits light having a wavelength matching the cell-specific fluorescent protein, and the fluorescence detector 13 detects the fluorescence intensity of the cells, and calculates the concentration of lead ions in the sample with reference to a previously prepared calibration curve.

Claims

1. A biotype heavy metal pollution detection device is characterized by comprising:

the sample/thallus co-incubation unit comprises a constant temperature tank, wherein one end of the constant temperature tank is provided with a water inlet, and the other end of the constant temperature tank is provided with a water outlet; a conical culture tube is arranged in the thermostatic bath, an opening is formed in the upper end of the conical culture tube, the bottom end of the conical culture tube is arranged in the middle of the bearing and is connected with a motor at the bottom of the thermostatic bath; the thermostatic bath is provided with an upper cover, the lower bottom surface of the upper cover is provided with a lug boss corresponding to each conical culture tube, and the lug boss correspondingly extends into the opening of each conical culture tube; the upper cover is provided with at least three insertion tubes corresponding to the conical culture tubes, the insertion tubes penetrate through the convex parts, one ends of the insertion tubes are inserted into the conical culture tubes, and the other ends of the insertion tubes are positioned outside the upper cover and are respectively connected with a culture medium storage tank, a buffer solution storage tank and a heavy suspension tube of the detection preparation unit through a pump and a pipeline;

2. The biological heavy metal pollution detection device according to claim 1, wherein one of the three insertion tubes is a central insertion tube, which is connected to the heavy suspension tube or the waste liquid tank through a pump and a pipeline, and a bottom tube opening end of the central insertion tube is lower than the other insertion tubes and faces a bottom center position of the conical culture tube.

3. The biotype heavy metal pollution detection device according to claim 1, wherein a temperature monitor and a heating rod are arranged in the constant temperature tank, and an insulating layer is arranged outside the constant temperature tank; the temperature monitor and the heating rod are in communication connection with the data processing and control module.

4. The biotype heavy metal pollution detection device of claim 1, wherein a circle of C-shaped groove is arranged on the peripheral side wall of the convex part of the upper cover, a ball is arranged in the C-shaped groove, and the ball is in contact with the inner wall surface of the conical culture tube.

5. The biotype heavy metal pollution detection device according to claim 1, wherein a perforated plate is provided on said thermostatic bath, said perforated plate is made of a heat insulating material, said perforated plate is provided with a plurality of through holes, said conical culture tubes are installed in said through holes and detachably connected to bearings at the bottom of said thermostatic bath; the opening of the conical culture tube is higher than the upper part of the pore plate.

6. The biological heavy metal pollution detection device according to claim 1, wherein the detection unit further comprises a socket for mounting the high transmittance glass detection vessel, the socket is provided with a plurality of slots, two opposite side surfaces of the slots are provided with windows for the light emitter to emit light to the high transmittance glass detection vessel, and the light is detected on the other side surface by the fluorescence detector.

7. The biological heavy metal pollution detection device according to claim 1, wherein the light emitter is controlled by the data processing and control module to emit light with two or more wavelengths in a switching manner; at least one ultraviolet light; the detection unit further comprises an absorbance detector for detecting the optical density of the bacterial liquid in the high-transmittance glass detection dish.

8. The biotype heavy metal pollution detection device according to claim 1, further comprising a housing, wherein the interior of the housing is divided into two layers, the upper layer is provided with the sample/thallus co-incubation unit and the detection preparation unit, and the lower layer is provided with the detection unit; the shell is also provided with a touch display screen.