Bacterial counting device and water treatment system

Abstract

The utility model provides a kind of bacterial counting device and water treatment system, it is related to the technical field of water quality microbiological detection device, sample pretreatment module includes filter assembly, and the water inlet end of filter assembly receives inlet water sample through pipeline, and the outlet end of filter assembly is connected with dyeing bin through pipeline, and pipeline intercommunication has drain pipe between filter assembly and dyeing bin, for the redundant water sample of outer discharge;CCD imaging module includes the imaging platform of pipeline intercommunication with the output end of dyeing bin, and automatic focusing assembly and fluorescence microscope are arranged on the upper side of the central axis of imaging platform, for showing the bacterial colony of fluorescent dye dyeing, and fluorescence microscope is electrically connected with CCD camera;CCD camera is electrically connected with image processing module, for outputting bacterial concentration and statistical distribution diagram, the technical problem of increasing culture time consumption and high detection cost of artificial operation in prior art, reaches fast detection, cost low and high accuracy technical effect.

Description

Technical Field

[0001] This utility model relates to the technical field of water quality microbial detection devices, and in particular to a bacterial counting device and a water treatment system. Background Technology

[0002] In industrial water treatment, such as reverse osmosis systems in thermal power plants, monitoring bacterial contamination in the water is crucial. During long-term water treatment in thermal power plants, when reservoir water is used as the source, the total bacterial count in the water is often abnormally high in summer. If the pretreatment system fails to effectively remove microorganisms, their entry into the reverse osmosis components can cause a rapid increase in the pressure differential, affecting system operating efficiency. Therefore, a highly efficient bacterial counting device is needed to assess the sterilization effect.

[0003] Currently, commonly used bacterial counting devices and methods are mainly divided into: plate culture method, which involves diluting water samples and spreading them on solid culture medium, incubating them in an incubator, and then manually counting the number of colonies. The device usually includes a culture dish, culture medium storage equipment, and a colony counter; flow cytometry, which uses a flow cytometer to perform high-speed analysis of single cells through fluorescent labeling and a fluid system. The device includes a laser light source, a flow cytometry channel, and a fluorescence detection module; and ATP (adenosine triphosphate) bioluminescence method, which uses an ATP detector to detect ATP content through a luciferase reaction to indirectly reflect the number of bacteria. The device is mostly a portable luminescence detection instrument.

[0004] However, while the plate culture method in existing bacterial counting devices has a simple structure, it relies on manual operation and requires an independent culture space, taking 24 to 48 hours, which is time-consuming and cannot achieve automated rapid detection. Although flow cytometers and ATP detectors can perform efficient analysis, their internal fluid channels and optical detection systems are complex, resulting in high hardware costs and the need for professional maintenance, leading to high detection costs. Utility Model Content

[0005] The purpose of this invention is to provide a bacterial counting device and a water treatment system to alleviate the technical problems of increased cultivation time and high detection costs caused by manual operation in the prior art.

[0006] In a first aspect, the present invention provides a bacterial counting device, comprising: a sample pretreatment module, a CCD imaging module, and an image processing module;

[0007] The sample pretreatment module includes a filtration assembly. The inlet end of the filtration assembly receives the incoming water sample through a pipeline, and the outlet end of the filtration assembly is connected to a staining chamber through a pipeline. A drain pipe is connected between the filtration assembly and the staining chamber to discharge excess water sample.

[0008] The CCD imaging module includes an imaging platform connected to the output pipe of the staining chamber. An autofocus component and a fluorescence microscope are arranged above the central axis of the imaging platform to display bacterial colonies stained with fluorescent dye. The fluorescence microscope is electrically connected to a CCD camera. The CCD camera is electrically connected to an image processing module to output bacterial concentration and statistical distribution maps.

[0009] Furthermore, the staining chamber includes an integrated DAPI storage chamber, a FITC storage chamber, and a PBS washing chamber;

[0010] The DAPI and FITC storage bins use black light-blocking materials to store DAPI fluorescent dyes and FITC fluorescent dyes, respectively.

[0011] The PBS washing chamber stores phosphate-buffered saline.

[0012] Furthermore, the DAPI storage module and the FITC storage module can be connected in series or in parallel.

[0013] Furthermore, the filter membrane pore size of the filter assembly is 0.20μm to 0.25μm.

[0014] Furthermore, the fluorescence microscope includes a violet light source in the 365nm–370nm wavelength range and a blue light source in the 488nm–500nm wavelength range.

[0015] Furthermore, the CCD camera is a back-illuminated CCD camera, and the resolution of the CCD camera is no less than 500 pixels.

[0016] Furthermore, the image processing module includes a background subtraction module, a threshold segmentation module, and a dual-channel signal overlap analysis module.

[0017] Secondly, the water treatment system provided by this utility model includes the above-mentioned bacterial counting device, and a sedimentation tank 4, an ultrafiltration device 5, an ultraviolet sterilization device 6, a reverse osmosis device 7, and a secondary desalination device 8 connected in sequence.

[0018] The bacterial counting device and the ultraviolet sterilization device 6 are connected in parallel, and water samples are taken from the inlet and outlet of the ultraviolet sterilization device 6.

[0019] Beneficial effects:

[0020] The bacterial counting device provided by this utility model includes: a sample pretreatment module including a filter assembly, the inlet end of which receives influent water sample through a pipeline, the outlet end of which is connected to a staining chamber through a pipeline, and a drain pipe connecting the filter assembly and the staining chamber for discharging excess water sample; a CCD imaging module including an imaging platform connected to the output end of the staining chamber through a pipeline, an autofocus assembly and a fluorescence microscope arranged above the central axis of the imaging platform for displaying bacterial colonies stained with fluorescent dye, and the fluorescence microscope being electrically connected to a CCD camera; and the CCD camera being electrically connected to an image processing module for outputting bacterial concentration and statistical distribution maps.

[0021] This invention integrates filtration, staining (DAPI / FITC dual dye stepwise injection), and washing functions through a sample pretreatment module to achieve automated concentration and labeling of bacteria; a CCD imaging module uses a dual LED light source of ultraviolet and blue light to excite fluorescence signals, combined with a high-resolution CCD camera to capture bacterial images; and an image processing module uses background correction, dual-channel fusion, and machine learning algorithms to accurately distinguish target bacteria from impurities and output bacterial concentration.

[0022] This invention solves the problems of time-consuming traditional culture methods and high cost of flow cytometry, and has the advantages of rapid detection, low cost and high accuracy. It is suitable for rapid on-site microbial monitoring of drinking water, food processing and medical wastewater. Attached Figure Description

[0023] To more clearly illustrate the specific embodiments of this utility model or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0024] Figure 1 This is a schematic diagram of the structure of the bacterial counting device provided in an embodiment of the present invention;

[0025] Figure 2 A schematic diagram showing the location of the bacterial counting device provided in an embodiment of this utility model in a thermal power plant water treatment system.

[0026] Icons: 1-Sample pretreatment module; 101-Filter assembly; 102-Staining chamber; 103-Drainage pipe; 104-DAPI storage chamber; 105-FITC storage chamber; 106-PBS washing chamber; 2-CCD imaging module; 201-Imaging platform; 202-Autofocus assembly; 203-Fluorescence microscope; 204-CCD camera; 3-Image processing module; 4-Sedimentation tank; 5-Ultrafiltration device; 6-Ultraviolet sterilization device; 7-Reverse osmosis device; 8-Secondary desalination device. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, and not all embodiments. The components of the embodiments of this utility model described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0028] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0029] It should be noted that similar labels and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.

[0030] In the description of this utility model, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship commonly used when the product of this utility model is in use. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model. In addition, the terms "first," "second," and "third," etc., are only used to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0031] Furthermore, terms such as "horizontal," "vertical," and "sag" do not imply that components must be absolutely horizontal or suspended, but rather that they can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal relative to "vertical," and does not mean that the structure must be completely horizontal, but can be slightly tilted.

[0032] In the description of this utility model, it should also be noted that, unless otherwise explicitly specified and limited, the terms "set," "install," "connect," and "link" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0033] The following detailed description, in conjunction with the accompanying drawings, outlines some embodiments of the present invention. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0034] like Figure 1 As shown, the bacterial counting device provided by this utility model includes: a sample pretreatment module 1, a CCD imaging module 2, and an image processing module 3;

[0035] The sample pretreatment module 1 includes a filter assembly 101. The inlet end of the filter assembly 101 receives the incoming water sample through a pipeline, and the outlet end of the filter assembly 101 is connected to the staining chamber 102 through a pipeline. A drain pipe 103 is connected between the filter assembly 101 and the staining chamber 102 for discharging excess water sample.

[0036] The CCD imaging module 2 includes an imaging platform 201 connected to the output pipe of the staining chamber 102. An autofocus component 202 and a fluorescence microscope 203 are arranged above the central axis of the imaging platform 201 for displaying bacterial colonies stained with fluorescent dye. The fluorescence microscope 203 is electrically connected to a CCD camera 204. The CCD camera 204 is electrically connected to the image processing module 3 for outputting bacterial concentration and statistical distribution maps.

[0037] Specifically, the inlet of the filter component 101 of the sample pretreatment module 1 is connected to an external water source through a pipeline, and its outlet is connected to the inlet of the staining chamber 102 through a pipeline. A drain pipe 103 is connected to the pipeline between the filter component 101 and the staining chamber 102 to drain excess water sample after filtration. The imaging platform 201 of the CCD imaging module 2 is connected to the output of the staining chamber 102 through a pipeline to receive the pretreated sample. Above the central axis of the imaging platform 201, an autofocus component 202 and a fluorescence microscope 203 are arranged sequentially from top to bottom. The eyepiece of the fluorescence microscope 203 is electrically connected to the CCD camera 204, and the CCD camera 204 is electrically connected to the image processing module 3 through a data cable.

[0038] The filter assembly 101 traps bacteria and concentrates the sample through a 0.22μm filter membrane, while the drain pipe 103 promptly discharges excess water sample. This process can concentrate low-concentration water samples by 10 to 20 times, solving the problem of insufficient detection sensitivity caused by the low number of bacteria in the water sample. The connection between the staining chamber 102 and the filter assembly 101 enables automated mixing and staining of the concentrated sample with DAPI (4',6-diamidinyl-2-phenylindole DNA dye) / FIT (fluorescein isothiocyanate DNA dye). Combined with 25℃ temperature control and 5 to 10-minute staining time control, this ensures that bacterial DNA and surface antigens are effectively labeled.

[0039] The imaging platform 201 allows pre-processed samples to be directly transported to the imaging area. The autofocus component 202 above the central axis is coaxially arranged with the fluorescence microscope 203. The fluorescence microscope 203 is equipped with three ultraviolet light sources and a blue light source, which excite the fluorescence signals of DAPI and FITC, respectively. The high-sensitivity CCD camera 204 captures dual-channel fluorescence images in real time and transmits them to the image processing module 3. Through background subtraction, Otsu threshold segmentation, and dual-channel signal overlap analysis algorithms, bacteria and impurities are distinguished, and the bacterial concentration and statistical distribution map are finally output. This achieves the technical effects of rapid detection, low cost, and low false detection rate.

[0040] In an embodiment of this invention, the staining chamber 102 includes an integrated DAPI storage chamber 104, a FITC storage chamber 105, and a PBS washing chamber 106.

[0041] DAPI storage compartment 104 and FITC storage compartment 105 use black light-blocking material to store DAPI fluorescent dye and FITC fluorescent dye, respectively.

[0042] PBS washing chamber 106 stores phosphate-buffered saline.

[0043] DAPI storage module 104 and FITC storage module 105 can be connected in series or in parallel.

[0044] The filter membrane of filter assembly 101 has a pore size of 0.20 μm to 0.25 μm.

[0045] Specifically, the staining chamber 102 integrates a DAPI storage chamber 104, a FITC storage chamber 105, and a PBS washing chamber 106. The DAPI and FITC storage chambers 104 and 105 are made of black light-shielding material to protect the fluorescent dye from light. They are connected to the staining chamber 102 in series or parallel via tubing, allowing for synchronous or stepwise dye injection according to staining requirements. The PBS washing chamber 106, also made of light-shielding material, stores PBS for cleaning impurities after staining. The filter assembly 101 contains a filter membrane with a pore size of 0.20 μm to 0.25 μm, preferably 0.22 μm.

[0046] The integration of dyeing chamber 102 realizes the integration of dye storage, automatic injection and cleaning functions. The series and parallel connection of DAPI storage chamber 104 and FITC storage chamber 105 supports step-by-step labeling of dual dyes (DAPI labeling first, then FITC labeling). Combined with temperature and time control, it can label live bacteria and eliminate interference from dead bacteria. The black light-blocking material prevents the dye from decomposing due to light, and the -20℃ storage temperature maintains the dye activity and ensures dyeing efficiency.

[0047] The filter membrane of filter component 101 effectively traps bacteria, solving the problem of insufficient detection sensitivity caused by the low number of bacteria in water samples.

[0048] In an embodiment of this invention, the fluorescence microscope 203 includes a violet light source in the 365nm-370nm wavelength range and a blue light source in the 488nm-500nm wavelength range.

[0049] CCD camera 204 is a back-illuminated CCD camera with a resolution of no less than 500 pixels.

[0050] Image processing module 3 includes a background subtraction module, a threshold segmentation module, and a dual-channel signal overlap analysis module.

[0051] Specifically, the fluorescence microscope 203 has a built-in violet light source in the 365nm-370nm band, preferably a blue light source in the 365nm and 488nm-500nm bands, preferably 488nm, which are used to excite DAPI and FITC fluorescent dyes, respectively; the objective lens of the fluorescence microscope 203 is arranged coaxially with the central axis of the imaging platform 201 to ensure that the fluorescence signal is incident perpendicularly.

[0052] The CCD camera 204 is back-illuminated with a resolution of no less than 5 megapixels. It is coupled to the eyepiece of the fluorescence microscope 203 via an optical interface to capture multi-channel fluorescence images in real time and transmit them to the image processing module 3.

[0053] The 365nm violet light source and the 488nm blue light source have the same wavelength matching for the excitation peaks of DAPI and FITC, which can maximize the fluorescence signal intensity and reduce background noise. The back-illuminated CCD camera 204 has a quantum efficiency of >80%, and with a 10× objective lens (NA=0.3), it can achieve a resolution of 1μm / pixel, clearly distinguishing the morphology of bacilli and cocci.

[0054] Image processing module 3 has a specific software algorithm that uses morphological filtering to eliminate non-specific fluorescent spots, uses threshold segmentation (Otsu algorithm) and morphological analysis to distinguish individual bacteria from impurities, and analyzes DAPI (blue) and FITC (green) signals to eliminate interference from dead bacteria or non-bacterial particles, and directly outputs bacterial concentration (CFU / mL) and statistical distribution map.

[0055] like Figure 2 As shown, this utility model also provides a water treatment system applied in the water treatment process of a thermal power plant, including the above-mentioned bacterial counting device and a sedimentation tank 4, an ultrafiltration device 5, an ultraviolet sterilization device 6, a reverse osmosis device 7, and a secondary desalination device 8 connected in sequence; the bacterial counting device and the ultraviolet sterilization device 6 are arranged in parallel, and water samples are taken from the inlet and outlet of the ultraviolet sterilization device 6.

[0056] Specifically, sedimentation tank 4 removes large particulate impurities from the water, ultrafiltration device 5 removes small particles, and ultraviolet sterilization device 6 kills bacteria in the water with ultraviolet light, preventing the subsequent reverse osmosis device 7 from becoming clogged due to bacterial growth. To quickly detect the sterilization effect in the water, a rapid bacterial counting device is connected in parallel with ultraviolet sterilization device 6.

[0057] Based on the above embodiments, the specific working process of the bacterial counting device provided by this utility model is as follows:

[0058] Water samples enter the microfiltration assembly 101, where the built-in filter membrane filters out target bacterial colonies, concentrating the water sample at a concentration ratio of 10–20 times. The concentrated water sample then enters the staining chamber 102. First, the micropump in the DAPI storage chamber 104 activates, controlling the flow rate at 1–5 mL / min and the staining time at 8–15 min, labeling all bacterial DNA. Second, the micropump in the FITC storage chamber 105 activates, controlling the flow rate at 1–5 mL / min and the staining time at 10–20 min, specifically binding to the target bacterial colony surface antigen. The two stages are switched via a valve to prevent cross-contamination. During this process, after the first and second stages, a high-frequency micropump in the PBS washing chamber 106 activates, rinsing with PBS solution to remove free dye three times per rinse at a flow rate of 0.5 mL.

[0059] After water sample pretreatment, it is fed into imaging platform 201 for automatic slide pressing. Fluorescence microscope 203 has ultraviolet and blue light sources, displaying bacterial colonies stained with two different fluorescent dyes. Autofocus component 202 adjusts the focus in real time based on image sharpness algorithm to select a suitable counting interface. High-sensitivity CCD camera 204 can clearly distinguish bacterial morphology (such as bacilli and cocci).

[0060] Image processing module 3 prioritizes processing recorded images, employing dark-field image subtraction and non-uniform illumination correction. It performs pixel-level fusion of DAPI (blue) and FITC (green) channel images, retaining only double-positive signal regions. Based on the Otsu algorithm, it dynamically sets the grayscale threshold and combines morphological opening operations (3×3 kernel) to separate adherent bacteria. Finally, it trains a convolutional neural network model to distinguish between real bacteria and fluorescent impurities. Based on the field of view area, sample volume, and number of spots, it automatically converts the data to CFU / mL or Cells / mL.

[0061] Based on the above embodiments, the detection efficiency of this utility model and existing counting devices and methods is shown in Table 1.

[0062]

[0063] Table 1

[0064] Cost comparison is shown in Table 2.

[0065]

[0066] Table 2

[0067] Performance parameters are shown in Table 3.

[0068]

[0069] Table 3

[0070] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and are not intended to limit it. Although the utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this utility model.

Claims

1. A bacterial counting device, characterized in that, include: Sample preprocessing module (1), CCD imaging module (2) and image processing module (3); The sample pretreatment module (1) includes a filter assembly (101). The inlet end of the filter assembly (101) receives the incoming water sample through a pipeline. The outlet end of the filter assembly (101) is connected to a staining chamber (102) through a pipeline. A drain pipe (103) is connected between the filter assembly (101) and the staining chamber (102) for draining excess water sample. The CCD imaging module (2) includes an imaging platform (201) connected to the output end of the staining chamber (102). An autofocus assembly (202) and a fluorescence microscope (203) are arranged above the central axis of the imaging platform (201) for displaying bacterial colonies stained with fluorescent dye. The fluorescence microscope (203) is electrically connected to a CCD camera (204). The CCD camera (204) is electrically connected to the image processing module (3) for outputting bacterial concentration and statistical distribution maps.

2. The bacterial counting device according to claim 1, characterized in that, The staining chamber (102) includes an integrated DAPI storage chamber (104), a FITC storage chamber (105), and a PBS washing chamber (106). The DAPI storage compartment (104) and the FITC storage compartment (105) use black light-blocking material to store DAPI fluorescent dye and FITC fluorescent dye respectively; The PBS washing chamber (106) stores phosphate-buffered saline.

3. The bacterial counting device according to claim 2, characterized in that, The DAPI storage bin (104) and the FITC storage bin (105) are connected in series or in parallel.

4. The bacterial counting device according to claim 1, characterized in that, The filter membrane of the filter assembly (101) has a pore size of 0.20 μm to 0.25 μm.

5. The bacterial counting device according to claim 1, characterized in that, The fluorescence microscope (203) includes a violet light source in the 365nm-370nm band and a blue light source in the 488nm-500nm band.

6. The bacterial counting device according to claim 1, characterized in that, The CCD camera (204) is a back-illuminated CCD camera, and the resolution of the CCD camera (204) is not less than 500 pixels.

7. The bacterial counting device according to claim 1, characterized in that, The image processing module (3) includes a background subtraction module, a threshold segmentation module, and a dual-channel signal overlap analysis module.

8. A water treatment system, characterized in that, include: The bacterial counting device according to any one of claims 1 to 7 and the sedimentation tank (4), ultrafiltration device (5), ultraviolet sterilization device (6), reverse osmosis device (7), and secondary desalination device (8) connected in sequence. The bacterial counting device is connected in parallel with the ultraviolet sterilization device (6) to extract water samples from the inlet and outlet of the ultraviolet sterilization device (6).

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