CN110823893A - Temperature control microscopic system for epidemiological analysis and operation method thereof - Google Patents
Temperature control microscopic system for epidemiological analysis and operation method thereof Download PDFInfo
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
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- G02B21/0008—Microscopes having a simple construction, e.g. portable microscopes
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Abstract
The embodiment of the specification discloses a temperature control microscope system for epidemiology analysis and an operation method thereof, which at least comprise the following steps: inverting the microscope and the precision temperature control slide; the accurate temperature control slide glass includes: the temperature sensing device comprises a glass slide substrate, a temperature sensing element, a patterned heating layer, a protective layer and a transparent protective cover; in the experiment, the temperature is flexibly and accurately controlled based on the patterned heating layer to observe the reactions of microorganisms in the sample liquid at different temperatures, and the sample liquid is protected by the transparent protective cover, so that the sample liquid is prevented from being polluted, and the condition that the microorganisms in the sample liquid spread to infect working personnel in the experiment is avoided; due to the existence of the transparent protective cover, the microbes in the sample liquid can be observed at a short distance based on an inverted microscope, so that a better observation effect is achieved. Furthermore, temperature treatment and sample preparation do not need to be carried out repeatedly, accurate temperature control is carried out in the observation process, the experiment means is flexible and diversified, the experiment result is more accurate, and the pathogenic bacteria infection risk caused by repeated sample preparation is avoided.
Description
Technical Field
The specification relates to the field of medical instruments, in particular to a temperature control microscope system for epidemiological analysis and an operation method thereof.
Background
Epidemics refer to infectious diseases that can infect a large population. Can be used for treating infectious diseases such as influenza, meningitis, cholera, etc. In epidemiological analysis, viruses, bacteria and other pathogenic microorganisms are observed by a microscope.
Currently, in the field of pathological analysis, a microscope is generally used for detection experiments. For example, the propagation process of microorganisms such as viruses and bacteria is observed by an inverted microscope to conduct therapeutic studies.
However, the propagation culture of microorganisms requires a suitable environment, and at present, the experimental observation can be performed only by using a microscope after the external culture, and the experimental observation does not have a function of controlling the temperature of the culture medium, and thus the reactions of microorganisms such as viruses and bacteria at different temperatures cannot be observed. There is an experimental method for observing the propagation process of bacteria or viruses by using an inverted microscope in the art, but the method and the device thereof also cannot observe the reaction of pathogenic microorganisms at different temperatures, and because of lack of effective isolation and protection, there are many cases in which scientific researchers are infected during the experimental process.
Disclosure of Invention
The purpose of the embodiments of the present specification is to provide a temperature control microscope system for epidemiological analysis and an operation method thereof, so as to effectively improve the accuracy and safety of microorganism observation experiments and the flexibility of experimental means.
In order to solve the above technical problem, the embodiments of the present specification are implemented as follows:
a temperature controlled microscopy system for epidemiological analysis comprising at least: the system comprises an inverted microscope and a precise temperature control slide used in cooperation with the inverted microscope; wherein the content of the first and second substances,
the precision temperature control slide comprises: the device comprises a glass slide substrate, a temperature sensing element, a patterned heating layer, a protective layer and a transparent protective cover, wherein the temperature sensing element and the patterned heating layer are positioned on one surface of the glass slide substrate and arranged in a sample detection area;
the surface of the glass slide substrate is also provided with a groove surrounding the sample detection area, the lower opening edge of the transparent protective cover is arranged in the groove so as to cover at least the sample detection area in the groove, and disinfectant is dripped into the groove during sample detection;
the patterned heating layer penetrates through the groove, extends out of two first contacts at the periphery of the sample detection area and is positioned at one end of the surface, and simultaneously, the temperature sensing element also extends out of two second contacts at one end of the surface; the two first contacts are used for being connected with the output end of the temperature control unit, and the two second contacts are used for being connected with the input end of the temperature control unit so as to control the sample liquid dripped in the sample detection area to reach the preset temperature according to the setting of a user.
Optionally, the method further comprises: electrically connecting the pressing block;
the lower surface of the electric connection pressing block is provided with electric connection salient points which are respectively corresponding to the first contact and the second contact, and the side surface of the electric connection pressing block is provided with a microscope interface which is connected with the inverted microscope;
after the electric connection pressing block is correspondingly placed on the glass slide substrate, the electric connection salient points are respectively contacted with the first contact and the second contact; the patterned heating layer and the temperature sensing element are respectively connected with a temperature control unit integrated in the inverted microscope through the microscope interface.
Optionally, the material of the patterned heating layer is ITO and/or metal.
Optionally, the patterned heating layer has a thickness in the range of 20-200 nm.
Optionally, the thickness of the temperature sensing element is equal to the thickness of the patterned heating layer.
Optionally, the protective layer is made of an insulating material, and the thickness of the protective layer is in a range of 20-1000 nm.
Optionally, the electrical connection compact is filled with a weight material for maintaining balance.
Optionally, the microscope interface of the electrically connected pressing block is a USB interface.
According to the technical scheme, the temperature can be flexibly and accurately controlled based on the patterned heating layer arranged in the accurate temperature control glass slide so as to observe the reactions of microorganisms in the sample liquid at different temperatures, the sample liquid can be protected by the transparent protection cover, the sample liquid is prevented from being polluted, and meanwhile, the condition that the microorganisms in the sample liquid are diffused to infect working personnel during experiments is avoided; because the observation of the sample in the epidemiological analysis can be carried out for several days, the scientific research personnel are in risk due to the failure of the disinfectant, the arrangement of the liquid storage tank enables the regular replacement of the disinfectant to be possible, the disinfectant in the liquid storage tank is sucked out at intervals of a certain time, and new disinfectant is supplemented, and the new disinfectant in the liquid storage tank can enter the groove again to continuously provide a protective zone for isolating viruses and bacteria; moreover, due to the existence of the transparent protective cover, the microorganisms in the sample liquid can be observed at a short distance based on an inverted microscope, and a better observation effect is achieved. Furthermore, analysis on the whole does not need to repeatedly carry out temperature treatment and sample preparation, and accurate temperature control can be carried out in the observation process, so that the experimental means is more diversified, the experimental result is more accurate, and the pathogenic bacteria infection risk caused by repeatedly preparing samples is avoided.
Drawings
In order to more clearly illustrate the embodiments of the present specification or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below, it is obvious that the drawings in the following description are only some embodiments described in the present specification, and for those skilled in the art, other drawings can be obtained according to the drawings without any creative effort.
Fig. 1 is a schematic view of a temperature-controlled microscope system for epidemiological analysis provided in the examples of the present specification.
Fig. 2 is a schematic structural diagram of a precision temperature-controlled slide provided in an embodiment of the present disclosure.
Fig. 3 is a second schematic structural view of the precision temperature-controlled slide provided in the embodiment of the present disclosure.
Fig. 4 is one of schematic structural diagrams of an electrically connected compact provided in an embodiment of the present specification.
Fig. 5 is a second schematic structural diagram of an electrically connected compact provided in an embodiment of the present disclosure.
Fig. 6 is a schematic structural diagram of a temperature sensing element provided in an embodiment of the present disclosure.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present specification, the technical solutions in the embodiments of the present specification will be clearly and completely described below with reference to the drawings in the embodiments of the present specification, and it is obvious that the described embodiments are only a part of the embodiments of the present specification, and not all of the embodiments. All other embodiments obtained by a person skilled in the art based on the embodiments in the present specification without any inventive step should fall within the scope of protection of the present specification.
Referring to fig. 1, a schematic view of a temperature control microscope system for epidemiological analysis provided in the embodiments of the present disclosure is shown, the temperature control microscope system at least includes: an inverted microscope 11, and a precision temperature controlled slide 12 for use with the inverted microscope 11. It should be understood that a signal connection can be established between the inverted microscope 11 and the precision temperature controlled slide 12, and that the precision temperature controlled slide 12 generally needs to be placed on the stage 111 of the inverted microscope 11 for use.
Referring to fig. 2 and 3, the precision temperature controlled slide 12 further includes: a slide substrate 121, a temperature sensing element 122, a patterned heating layer 123, a protective layer 124, and a transparent protective cover 125.
Wherein:
the slide substrate 121 is generally made of a transparent substrate, such as polymethyl methacrylate (PMM), commonly known as plexiglass; polystyrene PS; polycarbonate PC, and the like.
The patterned heating layer 123 is disposed on any surface of the slide glass substrate 121 and is disposed in the sample detection area, and its specific pattern may be set according to the process capability and experimental requirements, for example, it may be set as a wiring structure shown in fig. 2, in which the gap between adjacent wires may be set to be the same as the width of the wires or larger than the width of the wires, so as to avoid the adjacent wires from contacting and causing short circuit. In the embodiment of the present disclosure, the material of the patterned heating layer 123 may be ITO and/or metal, in other words, all wires in the patterned heating layer 123 may be transparent conductive ITO material, or all wires may be metal material, or one segment may be ITO material, and the remaining segment is metal material.
The temperature sensing element 122 is also located on the same surface of the slide substrate 121 as the patterned heater layer 123, and the temperature sensing element 122 is disposed within the sample detection region. Specifically, the temperature sensing element 122 may be disposed at a pattern gap of the patterned heating layer 123, preferably in a positional relationship as shown in fig. 2. Thus, there is no interference between the temperature sensing element 122 and the patterned heater layer 123, and the two may not cross to lead out independent wires. Wherein the temperature sensing element 122 is used for detecting the temperature of the sample liquid; the temperature sensing element 122 may be a conventional thin film temperature sensor, for example, as shown with reference to fig. 6, the thin film temperature sensing element includes at least: the substrate 122-1, the positive thermocouple membrane 122-2, the negative thermocouple membrane 122-3, the thermocouple junction 122-4, the pad membrane 122-5, the insulating transition membrane 122-6 and the protective membrane 122-7 have the effects of fast response time, small volume and accurate detection. In addition, reference may be made to other existing thin film temperature sensing elements.
The protective layer 124 covers at least the patterned heating layer 123 and the temperature sensing element 122, thereby functioning as an isolation insulator.
Meanwhile, the upper surface of the slide glass substrate 121 (i.e., the surface provided with the patterned heating layer 123 and the temperature sensing element 122 in fig. 2) is further provided with a groove 126 surrounding the sample detection region, and a transparent protective cover 125 is provided above the protective layer 124 with its lower opening edge placed in the groove 126 to cover at least the sample detection region therein, that is, the patterned heating layer 123 and the temperature sensing element 122 in the transparent protective cover 125. Thus, when the precise temperature control slide 12 is used for experimental observation, the transparent protection cover 125 does not need to be opened, and the sample liquid dripped on the precise temperature control slide 12 is observed by using an inverted microscope, so that the transparent protection cover 125 can prevent the sample liquid from being polluted and protect the purity of the sample liquid; meanwhile, the infection of scientific research personnel or testing personnel caused by the diffusion of viruses or bacteria in the sample liquid can be prevented, and particularly, a better protection effect is achieved under the condition that pathogenic bacteria are epidemic disease in the sample liquid level. Preferably, a disinfecting liquid is dropped into the groove during sample detection, for example, a disinfecting liquid is dropped 84, so that the sample liquid is sufficiently isolated in the transparent protective cover 125 through the groove.
In a preferred embodiment, the upper surface of the slide substrate 121 is provided with a liquid storage groove 127, and the liquid guide groove 129 is communicated with the liquid storage groove 127 and the groove 126. A flange 128 is arranged in a semi-surrounding manner at the upper opening of the liquid storage groove 127 and the liquid guide groove 129 so as to allow the upper limit of the liquid level in the liquid storage groove 127 and/or the liquid guide groove 129 to be slightly higher than the upper surface of the slide substrate 121; it will be appreciated that the liquid level in the reservoir 127 and/or the gutter 129 may not be planar due to the effect of the surface tension of the liquid, in which case the liquid level is convex, at least a portion of the liquid level is slightly above the upper surface of the slide substrate 121.
It should be understood that, when the sample liquid is dropped, the sample liquid should be dropped to a gap between the patterned heating layer 123 and the temperature sensing element 122, for example, a central region of a pattern (as a region framed by a dotted line) formed by the patterned heating layer 123 and the temperature sensing element 122, and the patterned heating layer 123 and the temperature sensing element 122 are not disposed in the region, and there is no shielding, so that the strain in the sample liquid can be observed through the objective lens of the inverted microscope.
The inverted microscope is used because the objective of the ordinary microscope is at the top, and because of the presence of the transparent protective cover 125, the objective of the ordinary microscope cannot be close enough to the sample liquid, and thus cannot achieve a high magnification for accurate observation, while the objective of the inverted microscope is at the bottom, and it can be close to the sample liquid from the bottom. Meanwhile, it is considered that bacteria or viruses to be observed in the sample liquid generally accumulate at the bottom. Therefore, in general, the inverted microscope with the objective lens below can observe bacteria or viruses based on the closest distance, and a better observation effect can be achieved.
It should be understood that in the present specification embodiment, the sample detection region may be a middle region of the slide substrate 121, and specifically, may range substantially to the region surrounded by the groove 126 in fig. 3.
The patterned heater layer 123 protrudes through the recess 126 at one end of the surface at the periphery of the sample detection area, for example, as shown in fig. 3, and protrudes at the right end of the surface of the slide substrate 121 at first contacts ① and ②, and at the same time, the temperature sensing element 122 also protrudes at one end of the surface at two second contacts, for example, as shown in fig. 3, and protrudes at the same end of the slide substrate 121 at second contacts ② and ②, wherein the two first contacts ② and ② are used to connect to the output of a temperature control unit (not shown), and the two second contacts ③ and ④ are used to connect to the input of the temperature control unit to control the sample liquid dropped in the sample detection area to a preset temperature according to user settings.
It should be understood that, in the present specification, the temperature control unit is not shown, and may be separately provided and separately control the patterned heating layer 123, or may be integrated in the inverted microscope, for example, to control the patterned heating layer 123 to heat through a human-machine interface of the inverted microscope.
By the temperature control microscopic system for epidemiological analysis, temperature can be flexibly and accurately controlled based on the patterned heating layer arranged in the accurate temperature control glass slide to observe the reaction of microorganisms in the sample liquid at different temperatures, the sample liquid can be protected by the transparent protective cover, the sample liquid is prevented from being polluted, and meanwhile, the condition that the microorganisms in the sample liquid are diffused to infect working personnel during experiments is avoided; moreover, due to the existence of the transparent protective cover, the microorganisms in the sample liquid can be observed at a short distance based on an inverted microscope, and a better observation effect is achieved. Furthermore, analysis on the whole does not need to be repeated for temperature treatment and sample preparation, and accurate temperature control can be performed in the observation process, so that the experiment means is more diversified, and the experiment result is more accurate. Meanwhile, the pathogenic bacteria infection risk caused by repeated preparation of samples is avoided.
In the embodiment of the present specification, referring to fig. 4 and 5, the temperature control microscope system may further include an electrical connection compact 13, wherein the lower surface of the electrical connection compact 13 is provided with electrical connection bumps 131 corresponding to the two first contacts ①② and the two second contacts ③④, four electrical connection bumps 131 are shown, each electrical connection bump 131 may be electrically connected to the internal bus by a wire or a spring, wherein the electrical connection bump 131 may be a conductive pillar that is nested in the slot of the aperture and can freely extend and retract, the electrical connection bumps 131 protrude out of a portion of the surface of the electrical connection compact 13 before the electrical connection compact 13 is placed on the precise temperature control slide 12, and the electrical connection bumps 131 are in contact connection with the first contacts and the second contacts respectively and compress back to a portion of the slot of the aperture after the electrical connection compact 13 is placed on the precise temperature control slide 12.
Meanwhile, the side surface of the electrical connection compact 13 is provided with a microscope interface 132 connected with the inverted microscope 11; if the temperature control unit is integrated inside the inverted microscope 11, the patterned heating layer 123 and the temperature sensing element 122 may be respectively connected with the temperature control unit integrated inside the inverted microscope 11 through the microscope interface 132. Preferably, the microscope interface 132 of the electrical connection pressing block 13 may be a USB interface, and is connected to the inverted microscope or the temperature control unit through a USB interface data line. .
Alternatively, in the present specification embodiment, the thickness of the patterned heating layer 123 may range from 20 to 200 nm.
Accordingly, in order to ensure surface flatness of the precision temperature control slide 12 and to simplify the process, the thickness of the temperature sensing element 122 is equal to the thickness of the patterned heating layer 123. It should be understood that the thickness of the patterned heating layer 123 and the thickness of the protective layer 124 in fig. 2 are not true to scale, and the exaggerated proportional relationship is adopted for the convenience of illustration, and the actual film thickness is much thinner than that shown in the figure.
Alternatively, the material of the protection layer 124 is an insulating material, such as silicon oxide, preferably silicon dioxide SiO2(ii) a The thickness of the protective layer 124 may range from 20-1000 nm.
Optionally, the electrical connection compact 13 is filled with a weight material for maintaining balance. Referring to fig. 4, the upper portion of the electrical connection compact 13 is filled with a weight 133, the material of the weight 133 may be metal or other high density material.
The temperature control microscope system for epidemiological analysis referred to above is described in detail below based on the usage scenario, specifically taking the example of observing the response of plague bacillus to temperature.
After the plague bacillus is cultured in a culture dish for 48 hours, collecting part of the plague bacillus from a culture solution as a sample solution to be dripped onto a precise temperature control slide glass, and particularly dripping the sample solution into a sample detection area covered by a transparent protective cover; referring to fig. 1, after the cover slip-covered precision temperature control slide 12 is placed on the stage 111 of the inverted microscope 11, the initial state is observed through the objective lens 112, and finally the cover slip is emitted to the component 113 through the optical path formed by the plurality of optical elements 114, wherein the component 113 may be an eyepiece so as to facilitate the staff to observe the plague bacillus in the sample solution, or the component 113 may also be a camera for shooting the reaction image of the plague bacillus in the sample solution.
It should be understood that the step of covering the cover glass is an optional step, and the cover glass may not be covered, and the sample liquid is directly dripped to the precise temperature control glass slide and then protected by the transparent protective cover, so that the purpose of safe observation can be achieved by the liquid.
Heating the sample liquid through a patterned heating layer on the precise temperature control glass slide, increasing the temperature to 37 ℃, keeping the temperature for 12 hours, observing the propagation condition, and shooting an image; and then raising the temperature to 70 ℃ again, keeping the temperature for 1 hour, then lowering the temperature to 37 ℃, keeping the temperature for 12 hours, observing the propagation condition, and shooting images. Scientific data of the high temperature resistance of the plague bacillus can be obtained.
After the experiment is finished, the temperature can be raised to 130 ℃ and kept for a certain time (the time is selected according to the type of the microorganism in the experiment, and can be preferably 10 minutes), and the virus, the bacteria and the like in the sample liquid are completely inactivated. In this way, the precision temperature controlled slide can be reused with medical items or discarded as a disposable medical item.
The embodiment provides an operation method of a temperature control microscope system, which comprises the following steps:
firstly, dropwise adding a sample solution into a sample detection area on the accurate temperature control glass slide, wherein the sample solution contains viruses and/or bacteria;
secondly, arranging the transparent protective cover on the accurate temperature control glass slide, and arranging the lower edge of the transparent protective cover in the groove;
thirdly, dropwise adding enough disinfectant into the liquid storage tank to enable the groove to be filled with the disinfectant;
fourthly, placing the accurate temperature control glass slide on an inverted microscope for observation, and controlling a temperature control unit by a user to enable the sample liquid to be at different temperatures;
fifthly, sucking out the disinfectant in the liquid storage tank and injecting new disinfectant after 3-12 hours after the third step; the step is repeatedly executed every 3 to 12 hours;
and sixthly, controlling the temperature control unit to enable the sample liquid to be heated to 130 ℃ and keeping for 10 minutes.
Compared with the prior art, when the temperature control microscope system in the embodiment of the specification is used for experimental observation of microorganisms such as viruses and bacteria, temperature treatment and sample preparation do not need to be carried out repeatedly, accurate temperature control can be carried out in the observation process, the experimental means are flexible and diversified, and the experimental result is more accurate. Moreover, the risk that the experiment operating personnel are infected by pathogenic bacteria caused by repeated sample preparation is avoided, and meanwhile, the sample liquid is protected by the transparent protective cover, so that the sample liquid is prevented from being polluted and the experiment operating personnel are prevented from being infected by diffusion.
The embodiments in the present specification are described in a progressive manner, and the same and similar parts among the embodiments are referred to each other, and each embodiment focuses on the differences from the other embodiments. In particular, for the system embodiment, since it is substantially similar to the method embodiment, the description is simple, and for the relevant points, reference may be made to the partial description of the method embodiment.
The above description is only an example of the present application and is not intended to limit the present application. Various modifications and changes may occur to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.
Claims (10)
1. A temperature-controlled microscopy system for epidemiological analysis, comprising at least: the system comprises an inverted microscope and a precise temperature control slide used in cooperation with the inverted microscope; wherein the content of the first and second substances,
the precision temperature control slide comprises: the device comprises a glass slide substrate, a temperature sensing element, a patterned heating layer, a protective layer and a transparent protective cover, wherein the temperature sensing element and the patterned heating layer are positioned on one surface of the glass slide substrate and arranged in a sample detection area;
the surface of the glass slide substrate is also provided with a groove surrounding the sample detection area, the lower opening edge of the transparent protective cover is arranged in the groove so as to cover at least the sample detection area in the groove, and disinfectant is dripped into the groove during sample detection;
the patterned heating layer penetrates through the groove, extends out of two first contacts at the periphery of the sample detection area and is positioned at one end of the surface, and simultaneously, the temperature sensing element also extends out of two second contacts at one end of the surface; the two first contacts are used for being connected with the output end of the temperature control unit, and the two second contacts are used for being connected with the input end of the temperature control unit so as to control the sample liquid dripped in the sample detection area to reach the preset temperature according to the setting of a user.
2. The temperature controlled microscopy system for epidemiological analysis of claim 1, further comprising: electrically connecting the pressing block;
the lower surface of the electric connection pressing block is provided with electric connection salient points which are respectively corresponding to the first contact and the second contact, and the side surface of the electric connection pressing block is provided with a microscope interface which is connected with the inverted microscope;
after the electric connection pressing block is correspondingly placed on the glass slide substrate, the electric connection salient points are respectively contacted with the first contact and the second contact; the patterned heating layer and the temperature sensing element are respectively connected with a temperature control unit integrated in the inverted microscope through the microscope interface.
3. The temperature control microscope system for epidemiological analysis according to claim 2, wherein the upper surface of the slide substrate is provided with a liquid storage tank, and the liquid guide tank is communicated with the liquid storage tank and the groove; the upper openings of the reservoir and the fluid-conducting bath are provided with a flange in a semi-surrounding manner to allow at least a portion of the liquid level in the reservoir 127 and/or the fluid-conducting bath 129 to be slightly higher than the upper surface of the slide substrate 121.
4. The temperature controlled microscopy system for epidemiological analysis according to any one of claims 1-3, wherein the patterned heating layer is made of ITO and/or metal.
5. The temperature control microscopy system for epidemiological analysis of any one of claims 1-3, wherein the patterned heating layer has a thickness in the range of 20-200 nm.
6. The temperature controlled microscopy system for epidemiological analysis of claim 5, wherein the thickness of the temperature sensing element is equal to the thickness of the patterned heating layer.
7. The temperature controlled microscopy system for epidemiological analysis according to any one of claims 1 to 3, wherein the protective layer is made of an insulating material and has a thickness in the range of 20 to 1000 nm.
8. The temperature controlled microscopy system for epidemiological analysis of claim 2, wherein the electrical connection compact is filled with a weight material for maintaining balance.
9. A temperature controlled microscopy system for epidemiological analysis as defined in claim 2 or 8, wherein the microscope interface of the electrically connected ballast is a USB interface.
10. A method of operating the temperature-controlled microscopy system as defined in claim 3, comprising the steps of:
firstly, dropwise adding a sample solution into a sample detection area on the accurate temperature control glass slide, wherein the sample solution contains viruses and/or bacteria;
secondly, arranging the transparent protective cover on the accurate temperature control glass slide, and arranging the lower edge of the transparent protective cover in the groove;
thirdly, dropwise adding enough disinfectant into the liquid storage tank to enable the groove to be filled with the disinfectant;
fourthly, placing the accurate temperature control glass slide on an inverted microscope for observation, and controlling a temperature control unit by a user to enable the sample liquid to be at different temperatures;
fifthly, sucking out the disinfectant in the liquid storage tank and injecting new disinfectant after 3-12 hours after the third step; the step is repeatedly executed every 3 to 12 hours;
and sixthly, controlling the temperature control unit to enable the sample liquid to be heated to 130 ℃ and keeping for 10 minutes.
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