A method for cleaning a slide sample staining reaction

Abstract

The application discloses a cleaning method for a slide sample dyeing reaction, and belongs to the technical field of improved methods and devices for dyeing tissues or cells on a slide, and comprises the following steps: a plurality of parallel beam-shaped cleaning reagent jet bodies are generated by using a nozzle to perform inclined spraying on the surface of a spraying area of the slide; the height of the nozzle, the distance between the nozzle and the slide, and the spraying angle and speed of the cleaning reagent jet body are adjusted; and a buffer distance is passed through, so that the fluid is collected and distributed to form a waterfall-shaped fluid, the sample is cleaned by flowing through the surface of the sample area, and the residual reagent in the previous step is removed. The application has the advantages of high cleaning efficiency, low cleaning reagent consumption, and low sample loss rate; by adjusting the pressure or the nozzle parameters, different cleaning intensity requirements can be quickly adapted; the structure is simple, the occupied space is small, various installation occasions can be adapted, and the application is suitable for popularization and use.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of improved methods and apparatus for staining tissue or cells on a slide, and more particularly to a method for washing a slide sample staining reaction. BACKGROUND

[0002] Immunohistochemistry is a technique for staining tissue or cells on a slide, through which the state of the tissue and cells in the sample can be observed under a microscope to diagnose a disease.

[0003] Generally, immunohistochemistry staining requires multiple reagents to be reacted on the sample step by step to achieve the staining of the sample. To ensure the smooth progress of each reagent, a washing step is required between each reagent to remove the excess reagent after the current reaction, so as to ensure that the next reagent can normally react with the detected tissue, thereby ensuring the quality of the staining.

[0004] The current manual staining process usually uses an intermediate reagent (such as a buffer) to rinse the reagent, and uses the method of pouring the buffer to remove the excess reagent of the current step, which can ensure clean washing, but consumes a large amount of reagent, and the flow rate of the pouring process cannot be controlled, which often directly splashes into the reaction area, which may damage or wash away the tissue or cells to be tested.

[0005] Another method for automatically implementing immunohistochemistry staining process by using an instrument is to remove the sample reagent (including the intermediate reagent used in the washing step) by using laminar flow replacement, specifically by using the method of adding liquid on one side and pumping liquid on the other side, so that the fluid flows by itself relying on capillary force or gravity along the slope. The flow rate is slow and needs to be repeated several times, which consumes less reagent, but the washing of the reagent is not thorough enough, and the residual reagent will lead to a decrease in the quality of the staining result. The reason may be that the flow rate of the fluid is too slow to fully exchange the liquid in a short time.

[0006] Therefore, it can be considered that the existing slide washing method is unbalanced between the fluid flow rate (i.e. the damage or loss rate of the sample) and the washing efficiency. Either the washing efficiency is high and the sample loss rate is large, such as centrifugal washing or manual pouring washing, or the sample loss rate is low, but the washing efficiency is low and the washing effect is general, such as the existing laminar flow replacement technology. The present application arises at the historic moment, and attempts to propose a new washing process with high washing efficiency, easy to control and without impact or damage to the biological sample of the slide. SUMMARY

[0007] The purpose of the present application is to solve the problems existing in the prior art, and to propose a method for washing a slide sample staining reaction.

[0008] The application adopts the following technical solutions:

[0009] A cleaning method for a glass slide sample dyeing reaction, comprising the following steps: generating a plurality of parallel beam-shaped cleaning reagent jet bodies by using a nozzle, adjusting the height of the nozzle, the distance between the nozzle and the glass slide, and the jet angle and jet speed of the cleaning reagent jet body, so that the cleaning reagent jet body is obliquely sprayed on the surface area outside the sample area of the glass slide, scattering and sputtering are generated, and after a buffer distance, the distributed fluid flows through the surface of the sample area to clean the sample to remove the residual reagents in the sample.

[0010] By using the above technical solution, the fluid forms an accelerated waterfall-like fluid on the glass surface through the close-packed hole spraying method, which is faster than the laminar flow generated by relying entirely on the self-flow of the fluid, so as to improve the cleaning efficiency of the existing laminar flow replacement cleaning technology.

[0011] The laminar flow replacement in the prior art, such as patent ZL03817566.5, can be understood as relying entirely on the self-flow of the fluid, or the existing cell laminar flow washing machine, which causes the laminar flow of the solution above the cells by means of liquid suction on one side and liquid suction on the other side in a short distance, so as to realize the cleaning method of solvent dilution and replacement. It is also difficult to apply to the cleaning of a wider glass slide area. The above two technologies are equivalent to cleaning the sample area under the laminar flow with a slow flow rate u0, which will not damage the sample area due to the slow flow rate. However, the slow flow rate causes slow solvent dissolution and exchange speed, so multiple repeated operations are required, and some stubborn residual reagents are often ineffective, and centrifugal cleaning is also required.

[0012] Preferably, the oblique spraying specifically refers to an included angle between the cleaning reagent jet body and the upper surface of the glass slide in the range of 10-60°, and the included angle range is further preferably 32±5°.

[0013] By using the above technical solution, the jet angle of the jet body, i.e. the included angle between the jet body and the surface of the glass slide 19, the smaller the jet angle, the smaller the sputtering amount and the sputtering distance, but the greater the impact force, so it is necessary to balance between the impact force and the cleaning efficiency. On the one hand, the relative stability of the waterfall-like fluid flowing along the slope is ensured, and on the other hand, it is also necessary to ensure that there is enough impact force to fully exchange the liquid on the sample surface.

[0014] Among them, the distributed fluid is in a flat flow state with an average flow rate u1, and the variation range of the fluid thickness and flow rate along the width of the glass slide is not more than 20%, and the average flow rate u1 is more than 20 times of the flow rate u0 of the static fluid on the 5° slope surface of the glass slide.

[0015] Further preferably, u1 is 40-400 times of u0. It is detected that the fluid belongs to a fluid mechanics transition flow or a low turbulence flow, and the turbulence Re is in the range of 2000-6000. The transition flow refers to the turbulence Re in the range of 2000-4000. It is detected that u0 is about 1.5 mm / s, that is, the static fluid of the slide glass with a length of 60 mm needs to flow from the top of the slope to the bottom at a speed of about 40 mm / s. According to the width of the slide glass of 107 mm, the fluid flowing through the sample is about 80-300 mm / s. Through the sample loss test, a faster speed such as about 200 mm / s can only cause sample loss, and the same is also a desirable range.

[0016] By adopting the technical scheme, the flow rate of the fluid is limited, the sample is uniformly washed by the accelerated waterfall fluid, and the cleaning efficiency is improved without impacting or damaging the sample.

[0017] In order to realize the above waterfall fluid state, the structure design of the nozzle is also proposed. The nozzle comprises a nozzle body and a mounting portion. The nozzle body is provided with a jet hole and a liquid inlet interface pipe. The mounting portion is provided with at least one strip-shaped hole for mounting the nozzle. A plurality of equidistantly and densely arranged jet holes form a dense hole structure. The side of the nozzle away from the jet hole is fixedly connected with the mounting portion. The two symmetrical side surfaces of the nozzle are respectively provided with one processing hole. Any processing hole is sealed by a plug.

[0018] By adopting the technical scheme, the nozzle with the dense hole structure is formed, a plurality of parallel beam-shaped cleaning reagent jet flows can be generated, and the jet angle of the cleaning reagent jet flow and the impact force on the surface of the slide glass can be controlled by adjusting the position of the nozzle and the slide glass, the opening position of the jet hole and the driving force of the liquid outlet. That is, by adjusting, testing and observing the structure and position of the nozzle, the fluid can more easily reach the flat waterfall state before reaching the sample area.

[0019] Further, the specific structure of the nozzle is that the nozzle body is a polygonal column cavity structure horizontally arranged. The top surface and the bottom surface of the nozzle body are parallel. The front surface of the nozzle body and the bottom surface form a chamfered surface. The chamfered surface is provided with at least one row of jet holes. The straight line where any row of jet holes is located is parallel to the horizontal plane, and the jet direction of the fluid in the jet hole is perpendicular to the chamfered surface.

[0020] The contact part between the liquid inlet pipe interface and the top surface of the nozzle body is subjected to full welding treatment, and the liquid inlet pipe interface is connected with the internal cavity of the nozzle body. The processing hole is used for drilling, reaming and chip removal of the internal cavity of the nozzle body. After processing, the plug is sleeved and sealed. There are two common sealing methods.

[0021] One is that the processing hole is used for drilling, reaming and chip removal of the nozzle body, the processing hole mouth is provided with a blocking ring, the plug is a rubber plug and is provided with a groove for filling the blocking ring to form a sealing structure;

[0022] Two is that the processing hole mouth is provided with a threaded wall, the plug is a hard metal, and the sealing treatment is performed by winding waterproof tape and screwing in; the mounting portion is welded on the upper part of the back of the nozzle body, and the top surface of the mounting portion is flush with the top surface of the nozzle body, and two strip-shaped holes are formed in the top surface of the mounting portion.

[0023] By adopting the above technical scheme, some structures of the nozzle are limited, and the following characteristics are obtained:

[0024] 1) The spraying direction can be directly controlled by chamfer surface processing, the top surface is controlled to be horizontal during installation, and various nozzle angles are processed according to requirements, and various nozzles are used to perform comprehensive tests to obtain the most suitable nozzle and washing process parameters;

[0025] 2) Single-row spraying and multi-row spraying screening: multi-row spraying aims to increase the fluid volume and impact force of the waterfall flow by multi-stage spraying, dispersion and sputtering back, but under the same liquid inlet volume and liquid inlet pressure, the water outlet pressure of each spraying hole is smaller than that of the single-row spraying mode, so it is necessary to prevent dripping.

[0026] 3) The mounting portion is fixedly connected to the upper part of the back of the nozzle body, and the top surface of the mounting portion is flush with the top surface of the nozzle body, and an integral molding processing structure can also be used, two strip-shaped holes are formed in the top surface of the mounting portion, and any one strip-shaped hole can be fixedly installed on the support by two or more screws, and the support can be a fixed support or a movable support.

[0027] Preferably, the inner diameter of the spraying hole is 0.1-1mm.

[0028] By adopting the above technical scheme, the diameter of the spraying hole is limited to a preferable range obtained through numerous spraying tests, and the diameter is too small to easily atomize, and the diameter is too large to easily produce a large sputtering range, thereby affecting the sample area.

[0029] The mounting structure of the nozzle has the following two modes:

[0030] One is fixed installation: any one strip-shaped hole can be installed on the fixed support by two or more screws, the fixed support is adjusted in angle, height and distance before being fixedly installed, the nozzle and the glass slide are fixedly arranged, and a fixed cleaning structure is formed;

[0031] Second, the movable installation: any one of the strip-shaped hole can be installed by two or more screws on the moving support, and the moving support is provided with a linear walking mechanism based on the fixed support, so that the nozzle can perform the pipeline cleaning mode on the linearly arranged glass slides.

[0032] By adopting the above technical scheme, when cleaning, the fixed cleaning can be selected for less samples, and the pipeline cleaning can be performed for more samples by arranging the glass slides in a linear manner and making the moving path of the moving support parallel to the arrangement path of the glass slides.

[0033] As an upgrading and strengthening cleaning solution, the liquid inlet interface of the nozzle is provided with a high-speed valve capable of being opened and closed at high speed.

[0034] By adopting the above technical scheme, the combination of the dense hole structure of the nozzle and the intermittent spraying mode can further enhance the cleaning effect.

[0035] To realize intermittent spraying, the application also provides a spraying control system, which comprises a high-speed valve for controlling the opening and closing of the nozzle, a reagent storage bottle connected to the liquid inlet pipe interface, a gas cylinder for pressurizing the reagent in the reagent storage bottle, a gas source power for controlling the pressure of the gas cylinder, and a control system for controlling the above elements.

[0036] By adopting the above technical scheme, the control system controls the opening and closing of the gas source power and the high-speed valve, and the reagent storage bottle provides liquid level feedback to the control system to remind the user to add liquid in time; the gas cylinder provides pressure feedback to the control system to remind the user to add gas in time, so as to control the spraying pressure of the cleaning reagent; the cleaning reagent with pressure forms a jet through the nozzle structure, and the high-speed valve realizes the intermittent generation of the jet from the nozzle.

[0037] In summary, the application has the following at least one beneficial technical effect:

[0038] 1. The application generates multiple parallel beam-shaped cleaning reagent jets through the nozzle, sprays the surface of the sample area of the glass slide obliquely, adjusts the height of the nozzle, the distance between the nozzle and the glass slide, and the spraying angle and speed of the cleaning reagent jet, and passes through a certain buffer distance to make the fluid gather and distribute to form a flat waterfall-shaped fluid state; the waterfall-shaped fluid flows through the surface of the sample area to clean the sample, and compared with the existing laminar flow replacement cleaning method, the flow rate is greatly improved, and under the condition of no sample loss, the dilution and exchange effect can be fully achieved, which has great advantages compared with the existing manual liquid pouring and other spraying methods.

[0039] 2. On the basis of the above, the application also proposes a combination of intermittent injection and close-packed hole injection. By controlling the frequency of the waterfall flow gap and the contact time, the sample area can be washed multiple times alternately, further reducing the continuous impact force on the sample area, while the cleaning efficiency is higher, the cleaning reagent consumption is less, and better staining effect can be achieved.

[0040] 3. In summary, the application has high cleaning efficiency and low cleaning reagent consumption. By adjusting the pressure or nozzle parameters, it can quickly adapt to different cleaning intensity requirements. The structure is simple, occupies less space, can adapt to various installation occasions, and is suitable for popularization and use. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 is a spraying mode and position schematic diagram of a cleaning method for slide sample dyeing reaction proposed by the application;

[0042] Figure 2 is a structure schematic diagram of a nozzle used in a cleaning method for slide sample dyeing reaction proposed by the application;

[0043] Figure 3 is a frame structure schematic diagram of a spraying control system used in a cleaning method for slide sample dyeing reaction proposed by the application;

[0044] Figure 4 is a perspective view of a spraying system provided by embodiment 5 of the application;

[0045] Figure 5 is a top view of a spraying system provided by embodiment 5 of the application;

[0046] Figure 6 is a side view of a spraying system provided by embodiment 5 of the application;

[0047] Figure 7 is a perspective view of a spraying adjustment mechanism in a spraying system provided by embodiment 5 of the application.

[0048] Explanation of reference signs: 1, nozzle; 101, nozzle body; 102, mounting portion; 103, strip-shaped hole; 104, long screw rod; 105, injection hole; 106, liquid inlet pipe; 107, plug; 2, mounting platform; 3, sliding support; 31, sliding groove; 32, inverted T-shaped leg; 33, screw rod; 34, driving motor; 35, bearing seat; 4, cushion block; 41, gasket; 5, first telescopic rod; 51, first hinged frame; 6, experimental platform; 7, cleaning reagent tank; 71, water outlet pipe; 72, electromagnetic valve; 73, air inlet pipe; 8, pressurizing pump; 9, support table; 10, sample table; 11, second telescopic rod; 111, second hinged frame; 12, sample tank; 13, drainage tank; 14, water receiving tank; 15, fixing pin; 16, bandage; 17, pull ring; 18, batten; 19, glass slide; 20, controller. DETAILED DESCRIPTION

[0049] The following will be described in detail in combination with the accompanying drawings. Figures 1-7 The present application is further described in detail.

[0050] Example 1

[0051] A cleaning method for glass slide sample dyeing reaction, comprising the following steps: generating a plurality of parallel beam-shaped cleaning reagent jet bodies by using a nozzle (1), adjusting the height of the nozzle (1), the distance between the nozzle (1) and the glass slide (19), and the injection angle and injection speed of the cleaning reagent jet body, so that the cleaning reagent jet body is obliquely injected to the surface area outside the sample area of the glass slide (19), scattering and sputtering are generated, and after a certain buffer distance, the distributed fluid flows through the surface of the sample area to clean the sample to remove the residual reagents in the sample.

[0052] It should be noted that:

[0053] I. Placement of the glass slide: it can be placed horizontally, 5-10° inclined, or even vertically, but the sample should be observed whether it slides off, and the fluid state should be observed, such as the fluid rolling speed is too fast, and extremely uneven in the width direction of the glass slide 19, forming a linear bundle-shaped fluid, which has a large impact force, similar to a turbulent flow shape, which is easy to take away the tissue or cells in the sample area, and the surface near the bundle-shaped fluid cannot be completely cleaned, and the transition flow state of the present application cannot be achieved, so the inclination of the glass slide surface needs to be reduced or even placed horizontally. Therefore, the reasonable placement of the glass slide 19 is the basis for the implementation of the present application.

[0054] II. The position of the spray area is set: the surface of the slide 19 is in turn a hand-holding area, a spray area and a sample area. The spray angle, i.e. the angle between the spray fluid and the slide 19, is controlled so that the hand-holding area is basically not splashed and remains dry, so that during the hand-holding operation, foreign matter on the hand, especially solvent, dust and dye, will not penetrate into the spray area and the sample area through capillary action of liquid wetting, avoiding the possibility of secondary pollution.

[0055] III. Multiple parallel beam-shaped cleaning reagent jet fluids: multiple jet fluids are arranged in an equidistant and dense arrangement, and the arrangement forms a spray surface parallel to the bottom edge of the slide 19, so that the spray intensity and spray angle of each jet fluid are basically the same along the width direction of the slide 19.

[0056] This step is achieved by the structure design of the spray head 1, which is as follows:

[0057] Referring to Figure 1 and 2 , the spray head 1 includes a spray head body 101, further includes a mounting portion 102 for mounting the spray head body 101, a plug 107 for plugging the processing hole of the spray head body 101, a liquid inlet interface pipe 106 and a plurality of spray holes 105 arranged in an equidistant and dense arrangement, and the plurality of spray holes 105 form a dense hole structure.

[0058] Further, the specific structure of the spray head 1 is as follows: the spray head body 101 is a polygonal column cavity structure laid horizontally, the top surface and the bottom surface of the spray head body 101 are parallel, and the front surface of the spray head body 101 specifically refers to one side surface facing the slide 19 forming a chamfered surface with the bottom surface, and the chamfered surface is provided with a row of spray holes 105; the row of spray holes 105 is parallel to the horizontal plane, and the spray direction of the spray hole 105 is perpendicular to the chamfered surface, which is convenient for directly controlling the spray direction through the processing of the chamfered surface; when mounting, the top surface is controlled to be horizontal as much as possible, and various spray head 1s with different spray angle chamfered surface angles are processed according to the needs, and various spray head 1s are used for comprehensive test to obtain the most suitable spray head 1 and spray washing process parameters, which mainly refer to the spray angle; the liquid inlet interface pipe 106 is arranged on the top surface of the spray head body 101 and communicates with the internal cavity of the spray head body 101; the two plugs 107 are symmetrically arranged at the processing holes of the two end surfaces of the spray head body 101 and are sealed by sealing glue + sealing ring; the mounting portion 102 is fixedly welded on the upper part of the back surface of the spray head body 101, and the top surface of the mounting portion 102 is flush with the top surface of the spray head body 101; alternatively, an integrated processing structure can be adopted, and two strip-shaped holes 103 are formed on the top surface of the mounting portion 102, and any one strip-shaped hole 103 can be fixedly mounted on the fixed support or the movable support by two or more screws.

[0059] V. Spray parameter adjustment, including the following limitations:

[0060] 1) height and distance of the spray hole: the difference between the height of any spray hole 105 to the contact point of the spray hole 105 to the surface of the slide 19 and the horizontal distance, the height of the spray hole is 20-40 cm, and the distance of the spray hole is 20-40 cm;

[0061] 2) diameter of the spray hole: the inner diameter of the spray hole 105 is 0.1-1 mm, too small is easy to atomize, too large is easy to produce a larger splash range, and then affect the sample area.

[0062] 3) distance between spray holes: the distance between the spray holes 105 is set to 2-4 mm, since the width of the slide 19 is 20-30 mm, considering that the jet body will have a tendency to spread, the width is about 2-2.5 mm, so it is necessary to gather each jet body together, and also to avoid a relatively thick beam-like turbulent body;

[0063] 4) spray speed: with a spray flow meter, the flow rate of each spray hole 105 is limited to 0.05-0.1 m 3 / h, the range is 1±0.2 m, and the gas pressure for driving the liquid to spray is 0.2-0.4 Mpa;

[0064] 5) spray angle: that is, inclined spraying, the spray angle of the jet body is specifically 10-60°. Further limited to 32±5°, that is, the angle between the jet body and the surface of the slide 19, the smaller the spray angle, the smaller the splash amount and the splash distance, but the greater the impact force, so it is necessary to balance between the impact force and the cleaning efficiency. On the one hand, it is necessary to ensure the relative stability of the waterfall flow when flowing along the slope, and on the other hand, it is also necessary to ensure that there is enough impact force to fully exchange the liquid on the sample surface.

[0065] 6) a certain buffer distance: the standard length of the slide 19 is 60-90 cm, the distance between the sample area and the handheld area is 10-20 cm, the buffer distance needs to be controlled to be 10-20 cm, and the splash distance of the jet body after contacting the slide 19 should not exceed 10 cm, preferably controlled within 7 cm, and after splashing, the jet body converges with the dispersed flow, and each dispersed flow converges with each other, which is directly observed to form a relatively uniform layered curtain-like flow state in the width direction. There is a buffer distance between the contact point of the jet body and the sample in the sample area. The jet body impacts and disperses on the slide 19 and produces splash, the adjacent dispersed flow and splash flow are fused, and gradually uniformly distributed through a certain buffer distance of the slope flow to form the final waterfall flow. The advantage of the waterfall flow is that the surface flows through a relatively thin thickness to save water, the flow is relatively gentle and will not cause impact to the sample, and the flow velocity presents a gradient distribution along the thickness direction, the lower the flow velocity is closer to the slide 19, thereby avoiding a relatively large impact force on the sample, and the relatively high flow velocity of the surface layer of the flow can fully play a dilution and exchange role, which has a great advantage compared with the existing manual liquid pouring and other spraying methods.

[0066] Example 2

[0067] This embodiment proposes a processing method of the nozzle 1:

[0068] Processing idea: the top surface, front surface, chamfered surface, bottom surface and back surface successively enclose a five-edged columnar cavity structure, each of the two end surfaces of the five-edged columnar cavity structure starts a processing hole, and after processing is completed, the plug 107 is used to plug the processing hole to ensure sealing.

[0069] Processing process:

[0070] 1) During processing, a square column material is cut to form a chamfered surface by cutting and polishing, and the angle of the chamfered surface is designed in advance;

[0071] 2) Then holes are drilled and expanded from both ends of the square column material to form a cavity, and threads are formed by tapping the hole wall, and a counterbore structure is formed in the processing hole for placing the cap portion of the plug 107;

[0072] 3) Drill the spray hole 105 on the chamfered surface, and process one or more rows of spray holes 105 according to requirements;

[0073] 4) Drill a hole in the center of the top surface and fixedly install the liquid inlet interface pipe 106 by welding or gluing;

[0074] 5) Fixedly weld the installation part 102 on the back surface.

[0075] Example 3

[0076] This embodiment designs a multi-row spray hole structure, and a plurality of parallel spray holes 105 are arranged on the chamfered surface of the nozzle body 101. The multi-row spray is intended to increase the fluid volume and impact force of the waterfall flow in stages through multi-stage spraying, dispersion and splash back, but under the same liquid inlet volume and liquid inlet pressure, the water outlet pressure of each spray hole 105 is smaller than that of the single-row spray mode, so it is necessary to prevent dripping.

[0077] Example 4

[0078] On the basis of Example 1, referring to Figure 3 , intermittent spraying of the special nozzle is realized by a spraying control system, and the high-speed valve is controlled by the spraying control system to switch the high-speed switch, the spraying control system includes the high-speed valve for controlling the switch of the special nozzle, the reagent storage bottle connected to the liquid inlet interface pipe 106, the gas cylinder for pressurizing the reagent in the reagent storage bottle, the gas source power for controlling the pressure of the gas cylinder, and the control system for controlling the above elements;

[0079] The control system controls the opening and closing of the gas source power (gas pump) and high-speed valve, and the reagent storage bottle provides liquid level feedback to the control system to remind timely liquid addition; the gas storage bottle provides pressure feedback to the control system to remind timely gas addition to control the jetting pressure of the cleaning reagent; the cleaning reagent with pressure forms a jet through the nozzle 1, and through the high-speed switching of the high-speed valve, the cleaning reagent can intermittently generate a jet from the nozzle 1.

[0080] The cleaning waterfall flow is in an intermittent flow mode, and the amount of fluid attached to the sample surface is the least during the intermittent flow, and the sample surface is washed again through a new round of waterfall flow during the continuous flow, so that each round of intermittent flow-continuous flow process is a reagent dilution exchange process. According to the dilution principle, under the condition of the same amount of dilution solvent, the more the dilution exchange times, the more the reaction reagents that can be carried, and the less the sample surface residue, so that the best cleaning effect is achieved under the condition of the minimum water consumption. However, the contact time of the waterfall flow with the sample during each continuous flow process must also be ensured, and the single washing effect of each continuous flow process must also be ensured. Therefore, the waterfall flow gap frequency and the contact time are two important factors for controlling the cleaning efficiency.

[0081] Comparative Example 1

[0082] Before cleaning, the glass slide was placed at an inclination of 5°, and the surface of the glass slide was sequentially a hand-holding area, a pouring area (corresponding to the spraying area of Example 1), and a sample area. The glass slide was placed on a beaker, and water was poured into the pouring area to perform water cleaning. The fluid flowing through the sample area was in a chaotic turbulent flow, and no continuous and thin waterfall-shaped fluid appeared.

[0083] Comparative Example 2

[0084] Before cleaning, the glass slide was placed at an inclination of 5°, and the surface of the glass slide was sequentially a hand-holding area, a middle area (corresponding to the spraying area of Example 1), and a sample area. A narrow water tank was arranged near the top of the hand-holding area, and a water outlet tank was arranged near the bottom of the sample area. Water was continuously added to the narrow water tank to gradually raise the water level and overflow to the hand-holding area. The entire surface of the glass slide was a relatively low-speed waterfall flow, and the test flow rate u0 was 1.52 mm / s.

[0085] The results of the comparative test are as follows:

[0086] Examples 1 and 4 and Comparative Examples 1 and 2 all use paraffin sections dyed by immunohistochemistry, which are subjected to the following drying and dewaxing treatment: (1) placing the paraffin tissue section on the sample area of the glass slide and placing it in a 65°C constant temperature box for about 1 h; (2) dewaxing: xylene I + 10 min→xylene II + 10 min→gradient alcohol (from high to low); then, using phosphate buffered saline (PB) as a cleaning reagent, the glass slide is cleaned.

[0087] The spray angle and position of the spray head need to be set and adjusted before cleaning in Example 1 and Example 4. The slide slope is set to 10°, the spray hole diameter is 0.5 mm, the spray hole spacing is 3 mm, the spray angle is 30°, and the spray head structure with the included angle between the chamfer surface and the bottom surface of the spray head body 1 is 130°.

[0088] The distance from each spray hole to the slide surface is the same, and the spray hole height and distance are adjusted, that is, the difference between the contact point of any spray hole to the slide surface and the height of the spray hole and the horizontal distance difference. The spray hole height is set to 20 cm, the horizontal distance is set to 30 cm, the inlet gas pressure is 0.4 Mpa, the flow rate is 0.036 m 3 / h, the range is 1 m, the inlet gas pressure is gradually reduced by 0.01 Mpa, the fluid in the sample area of the slide is gradually changed from a turbulent strip to a flat waterfall fluid state, and the average speed u1 of the waterfall fluid flowing through the sample area is randomly selected as 154 mm / s, which is 0.3 times u0, and is located in the range of 40-400 times, which still belongs to the transition fluid state. At this time, the inlet gas pressure is 0.102 Mpa.

[0089] Table 1: Effect of intermittent spraying on cleaning efficiency of the present application

[0090] In order to test the effect of different spraying frequencies on the waterfall fluid state, Example 4 is divided into 5 groups for testing, and the final result test is as shown in Table 1:

[0091]

[0092] In the cleaning process of Comparative Example 2, the sample cleanliness of the sample area cannot reach 99%, and the highest is only 95.1%, which affects the subsequent dyeing success, especially in multi-step reactions, and the intermediate reagents are all residual, which causes the risk of dyeing failure. As can be seen, after the combination of intermittent spraying and close-packed hole spraying, the impact on the sample is intermittent, so the loss rate is greatly reduced, and the dilution of the sample is also multiple washing, so the water consumption is greatly reduced.

[0093] Example 5

[0094] In order to realize the pipeline batch cleaning of the spray head 1, the embodiment provides a spraying system, which comprises a spraying adjusting mechanism for mounting the spray head 1, a liquid inlet control mechanism for liquid inlet of the spray head 1, and a sample placing mechanism for placing the slide 19; the spraying adjusting mechanism comprises a mounting platform 2 and a sliding support 3, and a cushion block 4 is arranged between the mounting part 102 and the mounting platform 2; the mounting part 102, the cushion block 4 and the mounting platform 2 are respectively provided with at least one strip-shaped hole 103; the strip-shaped hole 103 of any one of the mounting part 102 and the strip-shaped hole 103 of the cushion block 4 are penetrated by at least two long screws 104; the strip-shaped hole 103 of any one of the cushion block 4 and the strip-shaped hole 103 of the mounting platform 2 are penetrated by at least two long screws 104; the bottom surface of the mounting platform 2 is hingedly connected with at least three first telescopic rods 5, the bottom end of the first telescopic rod 5 is connected to the top surface of the sliding support 3, and an experimental platform 6 is arranged below the sliding support 3; the experimental platform 6 is provided with a sliding groove 31 matched with the sliding support 3.

[0095] The liquid inlet control mechanism comprises a cleaning reagent tank 7, an electromagnetic valve 72 and a pressurizing pump 8; the cleaning reagent tank 7 is arranged on the upper surface of the experimental platform 6; the bottom of the side wall of the cleaning reagent tank 7 is connected with a water outlet pipe 71, one end of the water outlet pipe 71 away from the cleaning reagent tank 7 is sealingly connected with a liquid inlet interface pipe 106, and the water outlet pipe 71 is further provided with the electromagnetic valve 72; the top surface of the cleaning reagent tank 7 is communicated with an air inlet pipe 73, and one end of the air inlet pipe 73 away from the cleaning reagent tank 7 is communicated with the pressurizing pump 8; wherein the controller 20 is a control system marked in the block diagram, the pressurizing pump 8 is a gas source power and is communicated with a gas storage bottle, the cleaning reagent tank 7 replaces the reagent storage bottle in the figure, so as to facilitate batch cleaning, and the electromagnetic valve 72 is a high-speed valve in the figure. Figure 3

[0096] The sample placing mechanism comprises a support table 9 and a sample table 10; the support table 9 is arranged on the upper surface of the experimental platform 6; the bottom surface of the sample table 10 is hingedly connected with a second telescopic rod 11, one end of the second telescopic rod 11 away from the sample table 10 is connected to the surface of the support table 9, and the upper surface of the support table 9 is further provided with a water receiving groove 14, and the length direction of the water receiving groove is parallel to the length direction of the screw rod 33; the upper surface of the sample table 10 is provided with a sample groove 12, the inner side wall of the sample groove 12 is provided with a batten 18, and every four battens 18 form a working position for placing the slide 19; the side surface of the sample table 10 close to the spray head 1 and the side surface of the sample table 10 away from the spray head 1 are symmetrical inclined surfaces, and the two inclined surfaces are respectively provided with a drainage groove 13 at equal distances; one end of the drainage groove 13 is communicated with the sample groove 12, and the other end of the drainage groove 13 is provided with the water receiving groove 14 directly below the outlet of the drainage groove 13, so as to drain the accumulated water in the sample groove 12 and collect the water by the water receiving groove 14.

[0097] ​When installing, first, the spacer 4 and the nozzle 1 are positioned together by a long screw rod 104, and then the spacer 4 is positioned in the strip-shaped hole 103 of the mounting platform 2 by another long screw rod 104. According to the horizontal distance requirement of the nozzle 1 and the slide glass 19, the position of the nozzle 1 is adjusted, the two long screw rods 104 are fastened, and the number of long screw rods 104 is appropriately increased according to the position of the nozzle 1, and the long screw rods 104 are adjusted to pass through several strip-shaped holes 103. When the nozzle 1, the spacer 4, and the mounting platform 2 are completely fastened together, the installation is completed.

[0098] To further tighten the three, refer to Figure 7 A non-slip washer 41 is arranged above the mounting portion 102, between the mounting portion 102 and the spacer 4, and between the spacer 4 and the mounting platform 2.

[0099] To prevent the slide glass 19 from moving during washing, refer to Figures 4-6 Two fixed pins 15 are symmetrically arranged on the two non-inclined side walls of the sample table 10, and a bandage 16 is fixedly connected to the two fixed pins 15 located on the two sides of the sample table 10. One pull ring 17 is arranged at each end of the bandage 16, the pull ring 17 is sleeved on the fixed pin 15 and is blocked by the cap of the fixed pin 15, and the bandage 16 is an elastic rope. The bandage 16 covers the edge part of the surface of the slide glass 19. The slide glass 19 is fixed by the batten 18 and the bandage 16. During cleaning, the slide glass 19 can be arranged only on the edge part of the surface of the slide glass 19 near the nozzle 1, so as to avoid blocking the waste liquid flowing out of the cleaned sample.

[0100] Fan scanning cleaning mode of the slide glass 19:

[0101] Before cleaning, the height and the spray angle of the nozzle 1 can be adjusted by the first telescopic rod 5, the height and the inclination angle of the slide glass 19 are adjusted by controlling the second telescopic rods 11, so as to adjust the height difference between the nozzle 1 and the slide glass 19, and control the basic included angle between the jet flow of the nozzle 1 and the surface of the slide glass 19.

[0102] The nozzle 1 is moved to correspond to the position of the slide glass 19 by driving the motor 34, one row of first telescopic rods 5 near the slide glass 19 is controlled to be stationary, and one row of first telescopic rods 5 far from the slide glass 19 is controlled to reciprocate, so as to periodically rotate the nozzle 1 and form a fan-shaped cleaning surface, thereby scanning and cleaning the surface of the slide glass 19.

[0103] Flow line cleaning mode of the slide glass 19:

[0104] Refer to Figures 1-3A linear translation driving mechanism is arranged on the sliding support 3 to drive the nozzle 1 to translate, so that batch samples can be cleaned in a pipeline. The bottom surface of the sliding support 3 is provided with a reverse T-shaped supporting leg 32 which is in sliding cooperation with the sliding groove 31. The side surface of the sliding support 3 is provided with a screw hole and is threadedly connected with a lead screw 33. One end of the lead screw 33 is connected with a driving motor 34. The other end of the lead screw 33 is connected with a bearing seat 35. The lead screw 33 is provided with smooth sections at both ends. One end of the smooth section is connected with the driving motor 34 through a shaft coupling. The other end of the smooth section is connected with the bearing hole of the bearing seat 35. The nozzle 1 can translate along the axial direction and the sliding direction through the sliding of the sliding support 3, so that multiple samples can be cleaned in a pipeline. The cleaning positions of the glass slides 19 are the same, so that the cleaning strength and the cleaning angle of the samples are basically the same. This effect cannot be achieved by other existing technologies.

[0105] The above are preferred embodiments of the present application, and are not intended to limit the protection scope of the present application. Any equivalent changes made on the basis of the structure, shape and principle of the present application shall be covered within the protection scope of the present application.

Claims

1. A method for cleaning of a slide sample staining reaction, characterized by: The method comprises the following steps: The multiple parallel beam-shaped cleaning reagent jet bodies are generated by the nozzle (1), the height of the nozzle (1), the distance between the nozzle (1) and the glass slide (19), and the jet angle and jet speed of the cleaning reagent jet body are adjusted, the surface area outside the sample area of the glass slide (19) is sprayed obliquely by the cleaning reagent jet body, scattering and sputtering are generated, and after a buffer distance, the distributed fluid is collected to form a flat cascading fluid state, the cascading fluid flows through the surface of the sample area to clean the sample by flowing, and the residual reagent in the sample is removed. The collected and distributed fluid is in a flat flow state with an average flow rate u1, and the variation range of the fluid thickness and flow rate along the width of the glass slide (19) is not more than 20%, the average flow rate u1 is compared with the flow rate u0 of the static fluid on the 5° slope surface of the glass slide (19), and u1 is more than 20 times of u0.

2. The method for cleaning a slide sample staining reaction according to claim 1, wherein: The oblique spraying is specifically that the included angle between the cleaning reagent jet body and the upper surface of the glass slide (19) is 10-60°.

3. A method for cleaning a slide sample staining reaction according to claim 2, wherein: The oblique spraying is specifically that the included angle between the cleaning reagent jet body and the upper surface of the glass slide (19) is 32±5°.

4. The method for cleaning a slide sample staining reaction according to claim 1, wherein: The nozzle (1) comprises a nozzle body (101) and a mounting portion (102), the nozzle body (101) is provided with a jet hole (105) and a liquid inlet interface pipe (106), and the mounting portion (102) is provided with at least one strip-shaped hole (103) for mounting the nozzle (1).

5. The method for cleaning a slide sample staining reaction according to claim 4, wherein: The side of the nozzle body (101) away from the mounting portion (102) is provided with at least one row of equidistantly distributed jet holes (105).

6. The method for cleaning a slide sample staining reaction according to claim 4, wherein, The inner diameter of the jet hole (105) is 0.1-1mm.

7. The method for cleaning a slide sample staining reaction according to claim 4, wherein, The liquid inlet interface pipe (106) is connected with a high-speed valve capable of being opened and closed at high speed.

8. The method for cleaning a slide sample staining reaction according to claim 7, wherein, The high-speed valve is electrically connected with a spraying control system, the spraying control system further comprises a reagent storage bottle connected with the liquid inlet interface pipe (106), a gas cylinder for pressurizing the reagent in the reagent storage bottle, a gas source power for controlling the pressure of the gas cylinder, and a control system for controlling the above elements; the control system controls the switching of the gas source power and the high-speed valve, and the reagent storage bottle provides liquid level feedback to the control system to remind timely liquid addition; the gas cylinder provides pressure feedback to the control system to remind timely gas addition, so as to control the jetting pressure of the cleaning reagent; the cleaning reagent with pressure forms a jet through the nozzle (1), and the high-speed switching valve realizes that the cleaning reagent can intermittently form a jet from the nozzle (1).

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