96-hole quantitative plate for detecting concentration of microorganisms in water

Abstract

The utility model discloses a 96-hole quantitative disc for detecting the concentration of microorganisms in water, the 96-hole quantitative disc comprises a composite layer, a transparent plate and a culture bin, the composite layer is of a multi-layer composite structure and comprises a metal barrier layer connected with the transparent plate, the metal barrier layer is used for blocking oxygen, water vapor and light, and the culture bin is used for culturing microorganisms. The cultivation bins are arranged on the surface, away from the composite layer, of the transparent plate and comprise the square cultivation bins and the long-strip-shaped cultivation bins which are arranged according to the cypress pine distribution law, and the total number of the square cultivation bins and the long-strip-shaped cultivation bins is 96. A filling port used for injecting a water sample and reagent mixed solution into the cultivation bin is formed between the composite layer and the transparent plate, and the filling port can be sealed by a sealing machine. According to the 96-hole quantitative plate for detecting the concentration of the microorganisms in the water, detection on the concentration of the microorganisms in the water can be simplified, the operation process is simple, operation is convenient, and the requirement for the professional degree of detection personnel can be effectively lowered.

Description

Technical Field

[0001] This utility model relates to the field of detection fixture technology, and in particular to a 96-well quantitative disc for detecting the concentration of microorganisms in water. Background Technology

[0002] Microorganisms in water, including coliform bacteria, Escherichia coli, and enteropathogenic bacteria, are important factors affecting water quality safety.

[0003] Traditional methods for detecting microbial concentrations in water mainly include culture methods and biochemical identification methods. Culture methods involve culturing bacteria on a specific culture medium and observing their growth to determine the presence of target bacteria. While this method is relatively accurate, it has a long testing cycle, typically requiring several days or even a week to obtain results.

[0004] Biochemical identification methods determine the species of bacteria by analyzing their biochemical characteristics, but the process is complex and requires specialized technicians and equipment.

[0005] Therefore, this application proposes a 96-well quantitative disc for detecting the concentration of microorganisms in water, which solves the problems of complex operation, high detection requirements and excessive manual operation in existing biochemical identification methods. Utility Model Content

[0006] This application provides a 96-well quantitative disc for detecting the concentration of microorganisms in water, comprising a composite layer, a transparent plate, and incubation chambers. The composite layer is a multi-layered composite structure and includes a metal barrier layer connecting the transparent plate. The metal barrier layer is used to block oxygen, water vapor, and light. The incubation chambers are disposed on the surface of the transparent plate away from the composite layer and include square incubation chambers and elongated incubation chambers arranged according to the Burson-Plains distribution law. The total number of square incubation chambers and elongated incubation chambers is 96. An injection port for injecting a mixture of water sample and reagent solution into the incubation chambers is provided between the composite layer and the transparent plate. The injection port can be sealed by a sealing machine.

[0007] In one possible implementation, the composite layer further includes a matte film layer and a moisture barrier layer, wherein the matte film layer is located on the outermost layer away from the transparent plate, and the matte film layer is connected to the metal barrier layer through the moisture barrier layer.

[0008] In one possible implementation, the moisture-proof layer is moisture-proof paper.

[0009] In one possible implementation, the metal barrier layer is tin foil.

[0010] In one possible implementation, the surface of the metal barrier layer is provided with hot melt adhesive for bonding the transparent plate, and the layers in the composite layer are bonded together by pressure bonding to form a flat composite sheet.

[0011] In one possible implementation, the square culture chamber and the elongated culture chamber are located on opposite sides, and the transparent plate has an anti-bending groove near the outer ring and at the connection between the square culture chamber and the elongated culture chamber, the anti-bending groove extending along the length of the elongated culture chamber.

[0012] In one possible implementation, the elongated culture chamber has a support groove at its center, the support groove being located on the side away from the transparent plate.

[0013] In one possible implementation, the longitudinal section of the support groove is trapezoidal, wherein the inner diameter of the bottom of the support groove is smaller than the inner diameter of the opening, and the longitudinal direction refers to the direction perpendicular to the transparent plate.

[0014] In one possible implementation, the length of the support groove is 1 / 7 to 1 / 5 of the total length of the elongated culture chamber.

[0015] Beneficial effects: Compared with the prior art, the 96-well quantitative disc for detecting the concentration of microorganisms in water provided in this application arranges the incubation chambers on a transparent plate according to Bosson's law of distribution. At the same time, an injection port is provided between the transparent plate and the composite layer. When detecting microorganisms in water, the water sample and reagent mixture can be directly added into the incubation chamber through the injection port. Then, the injection port is sealed with a sealing machine, and the microorganisms in the water can be directly observed and read after the incubation is completed. The operation process is simple and convenient, which can effectively reduce the professional requirements of the testing personnel and reduce manual operation.

[0016] These and other objects, features and advantages of this utility model will be fully realized through the following detailed description. Attached Figure Description

[0017] Figure 1 An exploded view of the 96-well quantitative disk used in this application for detecting the concentration of microorganisms in water is shown.

[0018] Figure 2 The exploded side view of the 96-well quantitative disk used in this application for detecting the concentration of microorganisms in water is shown.

[0019] Figure 3 A top view schematic diagram of the 96-well quantitative disk used in this application for detecting the concentration of microorganisms in water is shown.

[0020] Figure 4A graph showing the coliform count in water samples measured using the quantitative plate of this application is presented.

[0021] Figure 5 A graph showing the coliform count in water samples obtained using the same water sample via multi-tube fermentation is presented. Detailed Implementation

[0022] The following description is intended to disclose the present invention so that those skilled in the art can implement it. The preferred embodiments described below are merely examples, and other obvious variations will occur to those skilled in the art. The basic principles of the present invention defined in the following description can be applied to other embodiments, modifications, improvements, equivalents, and other technical solutions that do not depart from the spirit and scope of the present invention.

[0023] Those skilled in the art should understand that, in the disclosure of this specification, the terms "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. 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, the above terms should not be construed as limitations on this utility model.

[0024] It is understood that the term "a" should be understood as "at least one" or "one or more", that is, in one embodiment, the number of an element can be one, while in another embodiment, the number of the element can be multiple, and the term "a" should not be understood as a limitation on the number.

[0025] refer to Figures 1 to 5This application provides a 96-well quantitative plate for detecting the concentration of microorganisms in water, comprising a composite layer 10, a transparent plate 20, and incubation chambers 30. The composite layer 10 is a multi-layered composite structure and includes a metal barrier layer 11 connecting the transparent plate 20. The metal barrier layer 11 blocks oxygen, water vapor, and light. Preferably, the metal barrier layer 11 is tin foil, which serves as a thermally conductive electrolytic capacitor material and a flexible metal film. It not only blocks oxygen, water vapor, and light, but also provides good sealing properties due to the liquid sample, preventing leakage and ensuring the light-proof and heat-preserving requirements for microorganisms during incubation. The incubation chambers 30 are located on the surface of the transparent plate 20 away from the composite layer 10 and include square incubation chambers 31 and elongated incubation chambers 32 arranged according to Poisson's law. The total number of square incubation chambers 31 and elongated incubation chambers 32 is 96, representing the 96 wells of the quantitative plate. Furthermore, a filling port 401 for injecting a mixture of water sample and reagent solution into the culture chamber is provided between the composite layer 10 and the transparent plate 20. The filling port 401 can be sealed by a sealing machine. The mixture of water sample and reagent solution is injected into the square culture chamber 31 and the elongated culture chamber 32 through the filling port 401. Generally, the filling port 401 is located at one end along the length of the quantitative plate, for example, near the end of the elongated culture chamber 32.

[0026] The specific testing procedure is as follows:

[0027] First, prepare the instruments, equipment and reagents, including a programmable quantitative sealing machine, incubator, ultraviolet analyzer (optional, for Escherichia coli confirmation), quantitative detection tray (different sizes of detection trays can be prepared as needed), 100ml quantitative detection bottle, pipettes and tips, enzyme substrate reagents.

[0028] Then, select appropriate sampling points according to relevant standards and specifications to collect water samples. For different types of water sources (such as rivers, lakes, well water, tap water, etc.), select appropriate sampling methods and sampling frequencies according to their characteristics. The collected water samples should be sent to the laboratory for testing as soon as possible. If testing cannot be carried out in time, the water samples should be stored in a low-temperature (generally around 4°C) and light-protected environment, and the testing should be completed within the specified time.

[0029] Subsequently, water samples are treated by filtration to remove impurities, and water samples expected to contain high concentrations of coliform bacteria can be appropriately diluted.

[0030] Add the sample: Accurately pipette 100ml of water sample (if it is a diluted water sample, indicate the dilution factor) and slowly add it to a sterile 100ml quantitative detection bottle; use tweezers to pick up an appropriate amount of enzyme substrate reagent and carefully add it to the detection bottle containing the water sample. After tightening the cap, gently invert the detection bottle several times to ensure the enzyme substrate reagent and water sample are thoroughly mixed. Avoid vigorous shaking to prevent excessive air bubbles.

[0031] For dispensing and sealing, slowly pour the thoroughly mixed water sample and reagent solution into the 96-well quantitative pan through the filling port 401, preventing liquid splashing and air bubbles. Ensure that each well is filled and the liquid volume meets the requirements. Place the 96-well quantitative pan filled with the mixture in the corresponding position on the programmable quantitative sealing machine, start the sealing machine, and perform the sealing operation according to the operating instructions to ensure a tight seal and no leakage.

[0032] For incubation, place the sealed 96-well quantitative tray smoothly into an incubator preheated to 35-37℃. Close the incubator door and begin incubation. The incubation time is generally 24 hours. During incubation, avoid frequently opening the incubator door to prevent affecting the internal temperature stability. Simultaneously, carefully observe the incubator's operating status to ensure the temperature remains within the set range.

[0033] Result interpretation: After the culture is complete, carefully remove the quantitative detection plate and place it in a brightly lit place with a single-colored background for observation.

[0034] If the liquid inside the well is colorless, it is considered negative, meaning that no coliform bacteria were detected in that well.

[0035] If the liquid inside the cavity is yellow, it indicates that total coliforms or fecal coliforms have been detected.

[0036] To further confirm the presence of Escherichia coli, the quantitative detection plate can be observed under a UV analyzer. If fluorescence is produced simultaneously in the yellow holes, the result is considered positive for Escherichia coli.

[0037] Based on the number of positive wells, referring to the corresponding Most Probable Number (MPN) table and considering the dilution factor of the water sample, the coliform count in the water sample (expressed as MPN / 100ml) is calculated. Figure 4 The data in the table shown.

[0038] Experiments were conducted using a multi-tube fermentation method with the same water sample, such as... Figure 5 The data in the table shown.

[0039] Experimental comparison was conducted using paired variables of the logarithm of the technical solution in this application and the logarithm of the test tube method:

[0040] Based on the normal distribution principle, with a sample size of 20, 20 valid samples were prepared and divided into two groups. Paired t-tests were performed on the two methods to analyze whether there was a significant statistical difference between them. The data of the two methods were considered equivalent. The results are shown in the table below:

[0041] average value Number Ten tubes logarithm 1.821 20 This technique logarithm 1.828 20

[0042] Therefore, based on the experimental results, the results obtained by detecting the concentration of microorganisms in water using the technical solution of this application are basically consistent with those obtained by the traditional ten-tube method, indicating that the detection accuracy is basically the same. However, the operation process of the technical solution of this application has been greatly simplified, making the operation more convenient, while greatly reducing the number of employees involved and effectively reducing the professional requirements for testing personnel.

[0043] In one embodiment, the composite layer 10 further includes a matte film layer 13 and a moisture-proof layer 12, wherein the matte film layer 13 is located on the outermost layer away from the transparent plate 20, and can provide a matte frosted surface effect, making it easy to handle, while also reducing reflection and providing a soft visual effect. In addition, the matte film layer 13 is connected to the metal barrier layer 11 through the moisture-proof layer 12, wherein the moisture-proof layer 12 is preferably implemented as moisture-proof paper, which is easy to obtain and convenient to use.

[0044] In one embodiment, the surface of the metal barrier layer 11 is coated with hot melt adhesive for bonding the transparent plate 20, and the layers in the composite layer 10 are bonded together by pressing to form a flat composite sheet. Since the hot melt adhesive is solid at room temperature, it is heated to a certain temperature using a heating device to reach its melting point and transform into a liquid state. At this point, the liquid hot melt adhesive has good fluidity and wetting properties, allowing it to quickly fill the gaps between the adhered objects. As the temperature decreases, the hot melt adhesive gradually cools and solidifies, forming strong intermolecular forces that firmly bond the adhered objects together. This bonding method is convenient, fast, and possesses a certain degree of strength and durability.

[0045] In one embodiment, the square culture chamber 31 and the elongated culture chamber 32 are located on opposite sides. Meanwhile, the transparent plate 20 has an anti-bending groove 201 near the outer ring and at the connection between the square culture chamber 31 and the elongated culture chamber 32. The anti-bending groove 201 extends along the length direction of the elongated culture chamber 32, which can improve the structural strength and rigidity of the transparent plate 20 in this direction, i.e., the length direction of the transparent plate 20, and facilitate microbial culture experiments.

[0046] In one embodiment, the elongated culture chamber 32 is provided with a support groove 301 at the middle position, and the support groove 301 is located on the side away from the transparent plate 20. In this way, the support groove 301 makes the culture chamber less prone to deformation and makes the culture space of the culture chamber more stable.

[0047] Considering the large span of the elongated cultivation chamber 32, which poses a potential risk of structural instability, a more preferred design is that the longitudinal section of the support groove 301 is trapezoidal, wherein the inner diameter of the bottom of the support groove 301 is smaller than the inner diameter of the opening, and the longitudinal direction refers to the direction perpendicular to the transparent plate 20, thereby improving the structural stability of the elongated cultivation chamber 32.

[0048] In one embodiment, the length of the support groove 301 is 1 / 7 to 1 / 5 of the total length of the elongated culture chamber 32, preferably 1 / 6, which enables the elongated culture chamber 32 to be set up in a limited space while maintaining the structural stability of the elongated culture chamber 32.

[0049] Those skilled in the art should understand that the embodiments of the present invention described above and shown in the accompanying drawings are merely examples and do not limit the present invention. The advantages of the present invention have been fully and effectively realized. The functions and structural principles of the present invention have been shown and explained in the embodiments, and any modifications or variations may be made to the implementation of the present invention without departing from the stated principles.

Claims

1. A 96-well quantitative disc for detecting the concentration of microorganisms in water, characterized in that, The device includes a composite layer, a transparent plate, and incubation chambers. The composite layer is a multi-layered composite structure and includes a metal barrier layer connecting the transparent plate. The metal barrier layer is used to block oxygen, water vapor, and light. The incubation chambers are located on the surface of the transparent plate away from the composite layer and include square incubation chambers and elongated incubation chambers arranged according to Boehringer's law. The total number of square incubation chambers and elongated incubation chambers is 96. An injection port for injecting a mixture of water sample and reagent solution into the incubation chambers is provided between the composite layer and the transparent plate. The injection port can be sealed by a sealing machine.

2. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 1, characterized in that, The composite layer also includes a matte film layer and a moisture-proof layer, wherein the matte film layer is located on the outermost layer away from the transparent plate, and the matte film layer is connected to the metal barrier layer through the moisture-proof layer.

3. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 2, characterized in that, The moisture-proof layer is moisture-proof paper.

4. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 2, characterized in that, The metal barrier layer is tin foil.

5. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 4, characterized in that, The surface of the metal barrier layer is provided with hot melt adhesive for bonding the transparent plate, and the layers in the composite layer are bonded together by pressing to form a flat composite sheet.

6. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 1, characterized in that, The square culture chamber and the elongated culture chamber are located on opposite sides. The transparent plate has an anti-bending groove near the outer ring and at the connection between the square culture chamber and the elongated culture chamber. The anti-bending groove extends along the length of the elongated culture chamber.

7. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 1 or 6, characterized in that, The elongated culture chamber has a support groove in the middle, and the support groove is located on the side away from the transparent plate.

8. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 7, characterized in that, The longitudinal section of the support groove is trapezoidal, wherein the inner diameter of the bottom of the support groove is smaller than the inner diameter of the opening, and the longitudinal direction refers to the direction perpendicular to the transparent plate.

9. The 96-well quantitative disc for detecting microbial concentration in water as described in claim 7, characterized in that, The length of the support groove is 1 / 7 to 1 / 5 of the total length of the elongated cultivation chamber.

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