A filter membrane staining method and device for cell and bacteria detection

Abstract

The present invention discloses a filter membrane staining device for cell and bacteria detection, belonging to the field of cell and bacteria detection. The device comprises a workbench, a filter membrane cup holder disposed at the upper end of the workbench, a rubber seat connected to a negative pressure suction tube disposed on the filter membrane cup holder, a support disposed above the filter membrane cup holder, a liquid injection device disposed on the support, the support and the filter membrane cup holder moving relative to each other, the liquid injection device comprising an injection tube connected to a staining reagent kit, the injection tube connected to a heating device, a liquid adding tube disposed at the lower end of the heating device; and a pressing sampling device disposed at the upper end of the workbench for pressing the filter membrane cup into the rubber seat. The present invention aims to improve the efficiency and quality of staining.

Description

Technical Field

[0001] The present invention relates to the field of cell and bacteria detection, and in particular to a filter membrane staining method and equipment for cell and bacteria detection. Background Art

[0002] Smearing, staining, and preparing slides of cells or bacteria is a crucial step in medical testing and biological research. Existing methods for smearing, staining, and preparing slides range from manual smearing, staining, and preparing slides to fully automated procedures using instruments such as the Auto-CytePREP. Alternatively, manual smearing is performed with subsequent staining and preparation performed by instruments. These methods are often inefficient, complex, and require expensive equipment.

[0003] For this purpose, patent publication number CN102042924B discloses a method for staining and preparing cells or bacteria, which improves the existing technology to increase efficiency and reduce usage fees.

[0004] However, in the above technology, dyes such as carbolic acid fuchsin drip out from the liquid adding head and enter the detection container such as the sandwich cup through the liquid adding hole. In order to ensure the effect of acid-fast staining, the sandwich cup needs to be heated to 55-65 degrees Celsius. Because the sandwich cup is made of plastic parts with poor thermal conductivity, the external heating method has defects such as long heating time and uneven heating, which leads to uneven staining. Acid-fast bacteria and the like are poorly colored in the center of the filter membrane of the sandwich cup. Summary of the Invention

[0005] The object of the present invention is to address the above problems and provide a filter membrane staining method and equipment for cell and bacteria detection, which can improve the efficiency and quality of staining.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is: a filter membrane staining device for cell and bacteria detection, which includes a workbench, a filter membrane cup fixing frame is provided at the upper end of the workbench, a rubber seat connected to the negative pressure suction tube is provided on the filter membrane cup fixing frame, a support is provided above the filter membrane cup fixing frame, a liquid injection device is provided on the support, the support and the filter membrane cup fixing frame move relative to each other, the liquid injection device includes an injection tube connected to the staining reagent kit, the injection tube is connected to the heating device, and a liquid adding tube is provided at the lower end of the heating device, so that the dye added to the filter membrane cup is in a heated state to improve the staining efficiency and quality; at the same time, in order to avoid the occurrence of filter membrane damage and the like when negative pressure is applied, a pressing sampling device for pressing the filter membrane cup into the rubber seat is provided at the upper end of the workbench.

[0007] Furthermore, a structure of the heating device includes a heating chamber, which is a hollow body structure. A heating part is provided in the heating chamber, and the heating part can be an electric heating tube, etc.

[0008] Furthermore, a heat insulating cover is provided on the outside of the heating chamber to prevent heat loss and also to avoid affecting other instruments and equipment.

[0009] Furthermore, another structure of the heating device includes a spiral tube connecting the injection tube and the liquid adding tube, and the spiral tube is used to extend the movement distance of the dye and increase the amount of dye remaining in the heating device, ensuring that the dye is heated to the required temperature and ensuring that the heated dye reaches the required amount. The spiral tube is placed in the heating chamber, and a heating part is provided in the heating chamber. The heating part can adopt an electric heating tube, etc.

[0010] Furthermore, the heating chamber is filled with a heat-conducting medium to reduce uneven heating.

[0011] Furthermore, when the filter membrane cup is pushed downwards towards the rubber seat, if the filter membrane cup is tilted, the filter membrane may be damaged under negative pressure. For this reason, the pressing sampling device includes pressing parts respectively arranged on both sides of the rubber seat, and the pressing parts rotate with the rotating shaft to the top of the filter membrane cup and push the filter membrane cup downwards.

[0012] Furthermore, since the filter membrane cup and the rubber seat are in a tight fit after assembly, when the dyeing is completed and the inspector needs to remove the filter membrane cup, it will be difficult to separate the filter membrane cup from the rubber seat. For this reason, the rotating shaft is arranged horizontally, and a rotating roller is provided on the rotating shaft for driving the filter membrane cup to move up and down.

[0013] Furthermore, in order to facilitate the inspection personnel to remove the filter membrane cup and adapt to the outer diameter tolerance of the filter membrane cup, the rotating roller is coaxially arranged with the rotating axis, and an elastic area in contact with the outer wall of the filter membrane cup is provided on the outer side wall of the rotating roller.

[0014] Furthermore, in order to replace the rotating roller or adapt to filter membrane cups of different specifications, connecting parts are provided at both ends of the rotating roller, and the connecting parts are detachably connected to the rotating shaft.

[0015] Furthermore, in order to drive the rotating shafts to rotate synchronously and reduce the number of driving mechanisms, mutually meshing transmission gears are provided on the rotating shafts on both sides of the rubber seat, and one side of the rotating shaft is connected to the output shaft of the motor.

[0016] A method for staining a filter membrane using a filter membrane staining device for cell and bacteria detection comprises the following steps:

[0017] 1) Place the membrane cup in the plastic seat, ensuring that the membrane below the membrane cup fits in contact with the supporting mesh. Then, with the help of a delivery pump, the dye enters the heating device from the injection tube.

[0018] 2) The dye is heated to the required temperature in the heating device. When the liquid adding tube moves above the membrane cup, the valve on the liquid adding tube opens and the dye drips into the membrane cup. The amount of dye dripped is controlled by time, etc.

[0019] 3) After the dye is added dropwise and the dyeing reaction occurs, the suction system is activated, and the negative pressure pump connected to the negative pressure suction tube operates to extract the liquid in the filter membrane cup after the dyeing reaction. At this time, the cells, bacteria, etc. to be detected are retained on the filter membrane; according to the number of dyeing times required, the number of injection tubes and liquid adding tubes is arranged, and the relative movement of the filter membrane cup fixing frame and the support is supplemented to complete the steps of dye addition, extraction, secondary dye addition, extraction, etc. according to the established dyeing steps until the dyeing is completed;

[0020] 4) After staining is completed, remove the filter membrane cup from the gel holder.

[0021] The beneficial effects of the present invention are as follows: the dyeing solution and other reagents are directly heated, and the heated reagents are dripped onto the filter membrane. Since the filter membrane has a strong liquid adsorption capacity, after the reagents come into contact with the filter membrane, the dyeing solution and other reagents with a certain temperature can quickly come into contact with the specimen, thereby improving the efficiency of operations such as acid-fast staining; at the same time, since the filter membrane is easily stained when it is in the acid-fast staining solution environment for a long time, thereby interfering with the observation results, the method of directly heating the dyeing solution and other reagents can reduce the reaction time, thereby reducing the risk of coloration of the filter membrane, etc.

[0022] 1. The inspector places the membrane cup in the rubber seat, and then rotates the pressing plate toward the side of the membrane cup until the lower end of the pressing plate contacts the upper end of the membrane cup, thereby pushing the membrane cup downward until the filter membrane at the lower end of the membrane cup contacts the supporting mesh plate to ensure that the membrane will not be damaged during the dye extraction process.

[0023] 2. After dyeing is completed, the rotating shaft rotates under the drive of the motor, causing the rotating roller to push the filter membrane cup upward to facilitate the operator to remove the filter membrane cup. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram of the top view of the dyeing equipment.

[0025] Figure 2 It is a front view structural diagram of the dyeing equipment.

[0026] Figure 3 This is a schematic diagram of the heating device assembly of Example 2 (front view).

[0027] Figure 4 This is a schematic diagram of the heating device assembly of Example 3 (front view).

[0028] Figure 5It is a schematic diagram of the filter membrane cup fixing frame structure in a top view.

[0029] Figure 6 It is a structural diagram of the pressing part when it is working.

[0030] Figure 7 This is a schematic diagram of the structure when the pressing part is opened.

[0031] Figure 8 It is a schematic diagram of the assembly structure of the pressing part and the rotating shaft.

[0032] The text labels in the figure indicate: 1. workbench; 101. filter membrane cup; 2. filter membrane cup fixing frame; 3. rubber seat; 4. support; 5. liquid injection pipe; 6. liquid adding pipe; 7. heating chamber; 8. heating part; 9. heat insulation cover; 10. spiral tube; 11. heating chamber; 111. heating part; 12. pressing part; 13. rotating shaft; 14. rotating roller; 15. elastic area; 16. connecting part; 17. transmission gear; 18. motor. DETAILED DESCRIPTION

[0033] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is described in detail below with reference to the accompanying drawings. The description in this part is only exemplary and explanatory and should not have any limiting effect on the scope of protection of the present invention.

[0034] Example 1, as Figure 1-2 As shown, the specific structure of the present invention is a filter membrane staining device for cell and bacteria detection, which includes a workbench 1, a filter membrane cup fixing frame 2 is provided on the upper end of the workbench 1, a rubber seat 3 connected to the negative pressure suction tube is provided on the filter membrane cup fixing frame 2, the rubber seat 3 and the filter membrane cup 101 can adopt the rubber seat and filter membrane cup disclosed in the patent with publication number CN208420488U, the filter membrane at the lower end of the filter membrane cup 101 can adopt the microporous filter membrane disclosed in the patent with publication number CN106226896B, a support 4 is provided above the filter membrane cup fixing frame 2, a liquid injection device is provided on the support 4, the support 4 and the filter membrane cup fixing frame 2 move relative to each other, such as the support 4 is fixed, and the filter membrane cup fixing frame 2 moves laterally; or the filter membrane cup fixing frame 2 is fixed, and the support 4 moves laterally. In this embodiment, the support 4 is moved laterally, and the support 4 can be driven by a motor or a hydraulic drive, such as the movement method of the movable base described in the patent with publication number CN205898535U. At the same time, the method of connecting the injection tube to the staining reagent kit and the structure of the suction system can adopt the patent or select other existing technologies. They are not described in detail in this embodiment. The injection device includes an injection tube 5 connected to the staining reagent kit, and the injection tube 5 is connected to the heating device. The lower end of the heating device is provided with a liquid adding tube 6; the upper end of the workbench 1 is provided with a pressing sampling device for pressing the filter membrane cup 101 into the rubber seat 3.

[0035] Specific working process: place the filter membrane cup 101 vertically in the rubber seat 3, open the upper end of the filter membrane cup 101, and fit the filter membrane under the filter membrane cup 101 with the supporting mesh plate in the rubber seat 3. Then, under the action of the conveying pump, the dye enters the heating device from the injection tube 5.

[0036] The dye is heated to the required temperature in the heating device, and then the support 4 moves so that the liquid adding tube 6 moves to the top of the filter membrane cup 101. At this time, the valve on the liquid adding tube opens, and the heated dye drips into the filter membrane cup 101. The amount of dye added is controlled by time, etc.

[0037] After the dye is added and the dyeing reaction is completed, the suction system is started and the negative pressure pump connected to the negative pressure suction tube works to extract the liquid in the filter membrane cup 101. At this time, the cells, bacteria, etc. to be detected are left on the filter membrane; through multiple dyeing and negative pressure extraction, the dyeing is completed.

[0038] After dyeing is completed, the filter membrane cup 101 is taken out from the gel holder.

[0039] Example 2, as Figure 3 As shown, other structures and working methods of this embodiment can refer to Example 1. In this embodiment, the heating device includes a heating chamber 7, which is a sealed hollow body structure. Its upper and lower ends are respectively connected to the injection pipe 5 and the liquid adding pipe 6. The inner bottom wall of the heating chamber 7 is set as an inclined surface lowered toward the liquid adding pipe 6. The liquid adding pipe 6 is provided with a valve. A heating part 8 is provided in the heating chamber 7. The heating part 8 can adopt an electric heating pipe, a steam pipe or other device that can generate heat. This embodiment takes an electric heating pipe as an example. The outer side of the heating chamber 7 is provided with a heat insulation cover 9, which can be made of ceramic or other materials that are not easy to conduct heat.

[0040] During specific operation, the dye and the like enter the heating chamber 7 from the injection tube 5 and are heated to 55-65 degrees Celsius by the heating part 8. When the dye needs to be added, the valve on the liquid addition tube 6 is opened, and the heated dye flows out from the lower end of the liquid addition tube 6.

[0041] Example 3, as Figure 4 As shown, other structures and working methods of this embodiment can refer to Example 1. In this embodiment, the heating device includes a spiral tube 10 connecting the injection tube 5 and the liquid adding tube 6, and the spiral tube 10 is placed in the heating chamber 11. The heating chamber 11 is provided with a heating part 111. The heating part 111 can adopt an electric heating tube, a steam tube or other device that can generate heat. This embodiment takes the electric heating tube as an example. The heating chamber 11 is filled with a heat-conducting medium, and the heat-conducting medium can be water, oil, etc.

[0042] During specific operation, the dye and the like enter the spiral tube 10 from the liquid injection tube 5. At this time, the heating part 111 generates heat and works, so that the dye in the spiral tube 10 is heated to the required temperature. When the dye needs to be added, the valve on the liquid addition tube 6 is opened, and the heated dye flows out from the lower end of the liquid addition tube 6.

[0043] Example 4, as Figure 1 、 2 5, other structures and working methods of this embodiment can refer to Examples 1-3. The pressing sampling device includes rotating shafts 13 disposed on both sides of the rubber seat 3. The rotating shafts 13 are arranged along the moving direction of the support 4. Both ends of the rotating shafts 13 are connected to the bearing seat at the upper end of the workbench 1. The rotating shafts 13 on both sides of the rubber seat 3 are provided with mutually meshing transmission gears 17. One side of the rotating shaft 13 passes through the bearing seat and is connected to the output shaft of the motor 18. The motor 18 is arranged on the mounting seat.

[0044] A rotating roller 14 is provided on the rotating shaft 13, and the outer side wall of the rotating roller 14 contacts the side wall of the filter membrane cup 101. The surface of the rotating roller 14 is rough to increase the friction between the rotating roller 14 and the filter membrane cup 101. The filter membrane cup 101 is placed vertically on the rubber seat 3, and the rotating roller 14 is coaxially arranged with the rotating shaft 13. A pressing portion 12 extending along its tangent is provided on the outer side wall of the rotating roller 14. The pressing portion 12 rotates with the rotating shaft 13 to the top of the filter membrane cup 101 and pushes the filter membrane cup 101 downward.

[0045] During the specific operation: the inspector inserts the lower end of the filter membrane cup 101 into the adhesive seat 3. To increase the speed, the inspector does not need to install each filter membrane cup 101 in place. It only needs to be in a vertical state and not fall over. Then, the rotating shafts 13 on both sides of the filter membrane cup 101 rotate in opposite directions on the upper surface of one side of the filter membrane cup 101 under the drive of the motor 18, so that the pressing part 12 rotates toward the side of the filter membrane cup 101 until the pressing part 12 pushes the filter membrane on the filter membrane cup 101 to fit the support mesh plate in the adhesive seat 3, and then the motor 18 stops working. The motor 18 can be a servo motor, and the rotation angle of the rotating shaft 13 can be controlled in advance by parameters such as time, or a detection unit such as a pressure sensor or a contact switch can be set on the pressing part 12. When the detection unit contacts the filter membrane cup 101, the detection unit sends data to the control system that controls the operation of the motor 18. After the control system analyzes the data, it controls the motor 18 to stop.

[0046] After dyeing is completed, the output shaft of the motor 18 rotates in the opposite direction, so that the rotating shafts 13 on both sides of the filter membrane cup 101 rotate in opposite directions on the upper surface of one side of the filter membrane cup 101, so that the pressing part 12 rotates toward the far side of the filter membrane cup 101. At this time, the rotating roller 14 drives the filter membrane cup 101 to move upward to reduce the depth of the filter membrane cup 101 inserted into the glue seat 3, thereby making it easier for the inspector to remove the filter membrane cup 101.

[0047] Example 5, as Figure 6-8 As shown, other structures and working methods of this embodiment can refer to Example 4. The rotating roller 14 is coaxially arranged with the rotating shaft 13, but in this embodiment, the rotating roller 14 does not contact the filter membrane cup 101. An elastic area 15 in contact with the outer wall of the filter membrane cup 101 is provided on the outer wall of the rotating roller 14. The position of the elastic area 15 is approximately set opposite to the pressing part 12. The elastic area 15 can be made of rubber, silicone or other materials.

[0048] During specific operation: the pressing portion 12 rotates with the rotation shaft 13, pushing the filter membrane on the membrane cup 101 until it fits against the supporting mesh plate in the adhesive holder 3. When dyeing is completed, the rotation shaft 13 rotates until the elastic area 15 contacts the outer wall of the membrane cup 101, thereby driving the membrane cup 101 to rise. When the rotation shaft 13 completes its rotation, the pressing portion 12 is located obliquely below the membrane cup 101, and the elastic area 15 is separated from the membrane cup 101. Compared with Example 4, this embodiment can remove the membrane cup 101 with less effort, and is also convenient for placing a new membrane cup 101.

[0049] Example 6, as Figure 5 As shown, other structures and working methods of this embodiment can refer to Example 4 or 5, but in this embodiment, the rotating roller 14 is provided with connecting parts 16 coaxial therewith at both ends, and the rotating shaft 13 between adjacent connecting parts 16 is provided on the bearing of the bracket, and the bracket is provided at the upper end of the filter membrane cup fixing frame 2. Flanges are provided at the connection between the connecting part 16 and the rotating shaft 13, and the two adjacent flanges are connected by bolts.

[0050] During specific operation: when the elastic area 15 or the rotating roller 14 is worn to the point where it cannot push the filter membrane cup 101 to rise, or the outer diameter of the filter membrane cup 101 changes, the bolts connecting the rotating shaft 13 and the connecting part 16 are removed, and the required rotating roller 14 is replaced. After the replacement is completed, the bolts are re-tightened.

[0051] It should be noted that, in this article, the terms "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also includes other elements not explicitly listed, or also includes elements that are inherent to such process, method, article or apparatus.

[0052] This article uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only used to help understand the method of the present invention and its core ideas. The above is only a preferred implementation method of the present invention. It should be pointed out that due to the limitations of textual expression, there are objectively infinite specific structures. For ordinary technicians in this technical field, without departing from the principles of the present invention, they can make several improvements, modifications or changes, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the scope of protection of the present invention.

Claims

1. A filter membrane staining device for cell and bacteria detection, comprising a workbench (1), a filter membrane cup fixing frame (2) being provided at the upper end of the workbench (1), a rubber seat (3) connected to a negative pressure suction tube being provided on the filter membrane cup fixing frame (2), a support (4) being provided above the filter membrane cup fixing frame (2), a liquid injection device being provided on the support (4), and characterized in that: The support (4) and the filter membrane cup fixing frame (2) move relative to each other, and the liquid injection device includes a liquid injection tube (5) connected to the staining reagent kit, the liquid injection tube (5) is connected to the heating device, and the lower end of the heating device is provided with a liquid adding tube (6); the upper end of the workbench (1) is provided with a pressing sampling device for pressing the filter membrane cup (101) into the glue seat (3); the pressing sampling device includes a rotating shaft (13) disposed on both sides of the glue seat (3), the rotating shaft (13) is arranged horizontally, and a rotating roller (14) for driving the filter membrane cup (101) to move up and down is provided on the rotating shaft (13), the outer wall of the rotating roller (14) contacts the side wall of the filter membrane cup (101), and the outer wall of the rotating roller (14) is provided with a pressing portion (12) extending along its tangent, and the pressing portion (12) rotates with the rotating shaft (13) to the top of the filter membrane cup (101) and pushes the filter membrane cup (101) downward.

2. A filter membrane staining device for cell and bacteria detection according to claim 1, characterized in that: The heating device comprises a heating cavity (7), and a heating portion (8) is provided in the heating cavity (7).

3. A filter membrane staining device for cell and bacteria detection according to claim 2, characterized in that: The outer side of the heating chamber (7) is provided with a heat insulation cover (9).

4. The filter membrane staining device for cell and bacteria detection according to claim 1, characterized in that: The heating device comprises a spiral tube (10) connecting a liquid injection tube (5) and a liquid adding tube (6); the spiral tube (10) is placed in a heating chamber (11); and a heating portion (111) is provided in the heating chamber (11).

5. A filter membrane staining device for cell and bacteria detection according to claim 4, characterized in that: The heating chamber (11) is filled with a heat-conducting medium.

6. The filter membrane staining device for cell and bacteria detection according to claim 1, characterized in that: The rotating roller (14) is coaxially arranged with the rotating shaft (13), and an elastic area (15) in contact with the outer wall of the filter membrane cup (101) is provided on the outer side wall of the rotating roller (14).

7. The filter membrane staining device for cell and bacteria detection according to claim 1, characterized in that: Connecting portions (16) are provided at both ends of the rotating roller (14), and the connecting portions (16) are detachably connected to the rotating shaft (13).

8. The filter membrane staining device for cell and bacteria detection according to claim 1, characterized in that: Transmission gears (17) meshing with each other are provided on the rotating shafts (13) on both sides of the rubber seat (3), and one rotating shaft (13) is connected to the output shaft of the motor (18).

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