Centrifugal immunohistochemical staining method

Abstract

The centrifugal immunohistochemical staining method comprises the following steps that a slide is placed in a slide bin located on a centrifugal staining disc, a supporting part is arranged in the slide bin so that a gap can be formed between the bottom side of the slide and the bottom of the bin, and a section to be stained is attached to the bottom side of the slide; rotating the centrifugal dyeing disc, starting a dripping dyeing assembly, dripping reaction liquid towards one side of the slide by utilizing a dripping dyeing head at the tail end of the dripping dyeing assembly, and enabling the reaction liquid to flow to the section area; after dripping dyeing is completed, the centrifugal dyeing disc rotates at a high speed for a certain time, and the slide is directly spin-dried under the centrifugal effect. According to the invention, after the section is subjected to drop dyeing by the reaction liquid, the high-speed centrifugal principle is directly utilized, the centrifugal dyeing disc drives the surrounding slide to do centrifugal motion, so that the slide can be spin-dried, the steps of washing and drying by wind are not needed, and the method is essentially different from the existing method, so that the whole dyeing process steps are reduced, functional parts such as washing and blowing are omitted, and the cost is reduced. The dyeing machine is simpler and more compact in structure, and the dyeing efficiency is remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of tissue staining, and in particular to a centrifugal immunohistochemical staining method. Background Art

[0002] Tissue sections made of human tissue cells have no color themselves and cannot be identified under a microscope. In order to distinguish different cell structures, the sections must be stained. There are many methods for section staining, such as H1 staining, immunohistochemical staining, special staining, etc. General paraffin sections use the hematoxylin-eosin staining method. Hematoxylin can make the cell nucleus appear purple-blue, and eosin is an acid dye that mainly makes the components in the cytoplasm and extracellular matrix appear red. Different cell tissues and different proteins have different shades of color, which allows us to distinguish different types of tissue cells under a microscope. Most diseases can be correctly diagnosed by H1-stained sections. Sometimes H1 staining is difficult to confirm, and other staining methods need to be used to assist in diagnosis, such as immunohistochemical staining and special staining methods.

[0003] For example, the invention patent with authorization announcement number CN108663255B discloses a fully automatic immunohistochemical staining machine, whose basic working principle is: the sample needle can absorb the reagent and drip a small volume of reagent or staining liquid onto the tissue above the glass slide; the spray air-drying mechanism includes a spray needle and a blower fan. The spray needle is connected to multiple large-capacity reagent bottles, and through a pump or valve, according to preset rules, a large volume of reagent is sprayed onto the tissue on the glass slide; the blower fan is used to blow dry the reagent above the glass slide. When the mechanical arm assembly is working, when the spray needle of the spray air-drying mechanism sprays a large volume of reagent, the Y-axis linear module moves synchronously to ensure that the spray liquid can cover most of the glass slide; when the fan of the spray air-drying mechanism is turned on, the Y-axis linear module also moves synchronously to ensure that the blowing can sweep the entire glass slide to achieve the purpose of drying. The staining machine involved in this patent uses a movable spray air-drying mechanism for drying after staining. Summary of the invention

[0004] The purpose of the present invention is to provide a centrifugal immunohistochemical staining method, which uses a high-speed centrifugal method to process the stained glass, replacing the existing method of drying by air.

[0005] To achieve the above object, the present invention adopts the following technical solution:

[0006] A centrifugal immunohistochemical staining method comprises the following steps:

[0007] S1: placing a glass slide in a glass slide bin located on a centrifugal staining tray, wherein a support portion is provided in the glass slide bin to ensure a gap between the bottom side of the glass slide and the bin bottom, and a section to be stained is attached to the bottom side of the glass slide;

[0008] S2: The centrifugal staining disk rotates, the dripping dye assembly is started, and the reaction liquid is dripped toward one side of the glass slide using the dripping dye head at the end of the dripping dye assembly, and the reaction liquid flows toward the slice area;

[0009] S3: After the drop staining is completed, the centrifugal staining disk rotates at high speed for a certain period of time, and the glass slide is directly dried under the centrifugal effect.

[0010] Furthermore, the slide chambers are provided with a plurality of slide chambers distributed along the circumference of the centrifugal staining disk.

[0011] Furthermore, when placing the glass slide, one end of the glass slide is firstly abutted against the fixed limiting portion of the glass slide bin, and then the other end is abutted against the elastic limiting portion of the glass slide bin.

[0012] Furthermore, the elastic limiting portion includes a slider and a spring, one side of the slider is connected to the spring, and the other side opposite to the slider forms a limiting surface for the other end of the glass slide to abut against.

[0013] Furthermore, in step S1, the height of the support portion is set to 0.1-0.6 mm.

[0014] Furthermore, the centrifugal dyeing disk is located in the cylinder, the upper part of the cylinder is provided with the drip dyeing head, the drip dyeing assembly includes a liquid supply pipeline, a liquid pump and a reaction liquid bottle, and the liquid pump pumps the corresponding reaction liquid into the drip dyeing head.

[0015] Furthermore, the cylinder is connected to a hot air blowing pipe to blow hot air into the cylinder to keep the cylinder at a certain temperature.

[0016] Beneficial effects of the present invention:

[0017] After the slices are dyed with the reaction liquid, the invention does not rinse or blow dry the slides with wind, but directly uses the high-speed centrifugal principle, and the centrifugal staining disk drives the surrounding slides to do centrifugal movement to spin dry the slides. This is essentially different from the existing method, thereby reducing the steps of the entire dyeing process, simplifying the functional components such as rinsing and blowing, making the dyeing machine structure simpler and more compact, and improving the dyeing efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is an appearance diagram of a staining machine for implementing the centrifugal immunohistochemical staining method of the present invention;

[0019] Figure 2 yes Figure 1 The internal structure diagram of the dyeing machine shown;

[0020] Figure 3 This is a diagram of the structure of a centrifugal staining disk;

[0021] Figure 4 yes Figure 3A partial view of the figure (one cover is omitted).

[0022] The names corresponding to the marks in the figure are:

[0023] 1. Shell; 11. Touch screen; 12. Top cover; 21. Reaction liquid bottle; 22. Liquid pump; 23. Electric valve; 24. Dyeing head; 3. Cylinder; 41. Hot air blower; 42. Hot air blowpipe; 43. Temperature sensor; 5. Centrifugal staining plate; 51. Slide; 6. Slide bin; 61. Fixed limit part; 62. Elastic limit part; 621. Cover plate; 622. Sliding block; 623. Spring; 63. Support part; 7. Slide. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0025] Example:

[0026] To facilitate understanding of the dyeing method of the present invention, a dyeing machine capable of implementing the method is first introduced.

[0027] like Figure 1-Figure 4 As shown, the dyeing machine includes a shell 1, and a slope is provided on the front side of the shell 1. A touch screen 11 is provided on the slope, which can be connected to a control system. The operation of the dyeing machine can be controlled and some parameters, such as working time, internal temperature, etc., can be displayed by corresponding buttons on the touch screen 11.

[0028] Figure 2 and 3 As shown, a cylinder 3 is provided in the housing, a centrifugal staining disk 5 is provided in the cylinder 3, and a plurality of slide bins 6 are distributed along the circumference of the centrifugal staining disk 5. In this embodiment, 8 slide bins 6 are provided, and the number can be increased or decreased according to the situation in other embodiments. Figure 3 As shown, in order to facilitate understanding of the structural design, seven slides 7 are installed, and another slide chamber 6 is empty. A driving mechanism is provided at the bottom of the cylinder 3, including a motor, for driving the centrifugal staining disk 5 to rotate. This turntable structure is very mature and will not be described in detail. A top cover 12 is provided on the upper side of the housing 1, and one side of the top cover 12 is hinged. A handle is also provided on the top cover 12. The operator can hold the handle to flip the top cover 12 up and open it to perform related operations. The top cover 12 is closed during the staining process.

[0029] A dye dripping assembly is provided in the shell, and the dye dripping assembly includes a liquid supply pipeline, a reaction liquid bottle 21 , and a liquid pump. The liquid pump pumps the corresponding reaction liquid into the liquid spraying head 24 . Figure 2 In the structure shown, the dripping assembly includes six groups, each of which includes a reaction liquid bottle 21, a liquid pump 22, a liquid supply pipeline, and a dripping head 24. The dripping head 24 is arranged on the upper part of the cylinder 3, and can drip the reaction liquid toward the glass slide area in the cylinder 3, dripping 1 to 2 drops each time. Figure 2 The drip dye head shown in the figure is only a schematic diagram. The pipe of the drip dye head can be lengthened and positioned closer to the slide chamber, making it easier to perform the drip dye operation.

[0030] Each reaction liquid bottle 21 can be filled with a reaction liquid as needed for different immunohistochemical staining. For different tissue samples and detection purposes, one or more reaction liquids can be selected in combination with corresponding staining processes. The outlet of each pump is connected with an electric valve 23, such as a pipe clamp valve, for switching and regulating flow. The liquid pump adopts a micro pump to control the amount of reaction liquid each time. The present invention focuses on the staining process, so other mature reaction liquid injection mechanisms can also be used in other embodiments.

[0031] The glass slide bin 6 has a roughly rectangular trough body, which can accommodate a glass slide. A support portion 63 is provided on the bottom side of the trough body, and the bottom side of the placed glass slide is supported by the support portion 63, so that there is a gap between the bottom side of the glass slide and the bottom of the bin. The support portion 63 is provided with two groups, one group is a circular protrusion, and the other group is a strip-shaped protrusion along the length direction of the glass slide bin 6. The thickness of the support portion 63 is 0.1 to 0.6 mm, preferably 0.4 or 0.5 mm. The design of the support portion can meet the support of a certain height, but because the side of the glass slide with the sample is placed facing, the position of the support portion cannot affect the sample area of ​​the glass slide.

[0032] The outer end of the slide chamber 6 is provided with a fixed stopper 61, and the inner end (the end on the centrifugal staining disk 5) is provided with an elastic stopper 62. When placing the slide, the side with the sample faces upward, and one end of the slide 7 is firstly abutted against the fixed stopper 61 of the slide chamber 6, and then the other end is abutted against the elastic stopper 62 of the slide chamber 6, and the slide 7 is clamped by the elastic stopper 62 to prevent it from falling off during centrifugal movement.

[0033] Figure 3 and 4 As shown, the elastic limiting portion 62 includes a slider 622 and a spring 623. One side of the slider 622 is connected to the spring 623, and the other side opposite to the slider forms a limiting surface for the other end of the glass slide 7 to abut against. The slider 622 is L-shaped, with one end extending vertically into the glass slide bin 6, and the other end being horizontally arranged and connected to one end of the spring 623. A slide groove 51 is provided on the centrifugal staining tray 5 for the horizontal end of the slider 622 to extend into, and the spring 623 is located in the slide groove 51. When the slider 622 is subjected to force, it can move in the length direction of the glass slide bin 6. A cover plate 621 is also provided on the upper side of the slide groove 51, and the cover plate 621 is used to cover the spring 623 area. The cover plate 621 is fixed to the upper side of the centrifugal staining tray 5 by screws, and the cover plate is numbered with Arabic numerals.

[0034] Figure 2As shown, the dyeing machine is also equipped with a drying module, which can be used as an option. The drying module includes a hot air blower 41 and a hot air blowpipe 42 connected thereto. One end of the hot air blowpipe 42 is arranged on the cylinder 3, and can blow hot air toward the cylinder 3. During the dyeing process, the cylinder 3 is in a certain suitable temperature range, which is conducive to improving the dyeing effect. The temperature sensor 43 installed on the cylinder 3 is used to measure the temperature inside the cylinder 3 and feed it back to the control system to adjust the hot air blower 41.

[0035] The present invention provides a centrifugal immunohistochemical staining method, which can be implemented using the above-mentioned staining machine, comprising the following steps:

[0036] S1: Place a glass slide in a glass slide bin located on a centrifugal staining tray. A support portion is provided in the glass slide bin to ensure a gap between the bottom side of the glass slide and the bin bottom. The bottom side of the glass slide is attached with a section to be stained.

[0037] The side of the slide coated with the sample is facing downward. In the present embodiment, the sample is selected as a frozen section, and the sectioning and fixing on the slide are all processed according to the prior art.

[0038] When placing the glass slide, one end of the glass slide is firstly abutted against the fixed limiting portion of the glass slide bin, and then the other end is abutted against the elastic limiting portion of the glass slide bin.

[0039] S2: The centrifugal staining disk rotates, the dripping component is started, and the reaction liquid is dripped toward one side of the glass slide using the dripping head at the end of the dripping component, and the reaction liquid flows toward the slice area.

[0040] This side refers to the gap between the glass slide and the side wall of the trough of the slide bin. When dripping from top to bottom, due to the small interval of a few tenths of a millimeter between the glass slide and the bottom of the trough of the slide bin, a siphon effect can occur on the lower side of the glass slide, and the reaction liquid can quickly disperse along the surface of the glass slide to achieve the purpose of staining the slice.

[0041] The reaction solution may also be called a staining solution or a reagent, which is actually a certain type of antibody that reacts with a specific antigen in the slice. The antibody may be labeled in a manner known in the art.

[0042] The number of dripping heads can be set according to needs, such as 8 distributed along the circumference, which just corresponds to the glass slide bins one by one, and can drip dye at the same time. It is also possible to set 4 dripping heads. After dripping dye on four glass slides, it is necessary to rotate the centrifugal staining disk at a certain angle to drip dye on another glass slide again. The centrifugal staining disk can be driven by a servo motor to facilitate the control of the rotation angle and speed.

[0043] During the dyeing process, the hot air blower can be selectively turned on to keep the dyeing environment at a constant temperature.

[0044] S3: After the dripping is completed, the centrifugal staining disk rotates at high speed for a certain period of time, and the glass slide is directly dried under the centrifugal effect. Since there is a gap between the lower side of the glass slide and the bottom of the glass slide bin, and it is connected to the external space, the reaction liquid can be quickly thrown out during high-speed centrifugal movement.

[0045] When finished, push the slide toward the center of the centrifugal staining dish by hand to remove the slide.

[0046] In this embodiment, the glass slide is not rinsed or blown dry by wind, but directly uses the principle of high-speed centrifugation. The design is ingenious and original, which is obviously different from the existing dyeing process. The dyed glass slide is directly dried by centrifugation, which can reduce the steps of the entire dyeing process and simplify the functional components such as rinsing and blowing. The dyeing machine structure is simpler and more compact, and the dyeing efficiency is improved.

[0047] The present invention is not limited to the above-mentioned optimal implementation mode. Anyone can derive other various forms of products under the inspiration of the present invention. However, no matter what changes are made in the shape or structure, all technical solutions that are the same or similar to those of the present application fall within the protection scope of the present invention.

Claims

1. A centrifugal immunohistochemical staining method, characterized in that: The following steps are included: S1: placing a glass slide in a glass slide bin located on a centrifugal staining tray, wherein a support portion is provided in the glass slide bin to ensure a gap between the bottom side of the glass slide and the bin bottom, and a section to be stained is attached to the bottom side of the glass slide; S2: The centrifugal staining disk rotates, the dripping dye assembly is started, and the reaction liquid is dripped toward one side of the glass slide using the dripping dye head at the end of the dripping dye assembly, and the reaction liquid flows toward the slice area; S3: After the drop staining is completed, the centrifugal staining disk rotates at high speed for a certain period of time, and the glass slide is directly dried under the centrifugal effect.

2. The centrifugal immunohistochemical staining method according to claim 1, characterized in that: The slide chambers are provided with a plurality of slide chambers distributed along the circumference of the centrifugal staining disk.

3. The centrifugal immunohistochemical staining method according to claim 1, characterized in that: When placing the glass slide, one end of the glass slide is firstly abutted against the fixed limiting portion of the glass slide bin, and then the other end is abutted against the elastic limiting portion of the glass slide bin.

4. A dry dyeing method according to claim 3, characterized in that: The elastic limiting part comprises a slider and a spring. One side of the slider is connected to the spring, and the other side opposite thereto forms a limiting surface for the other end of the glass slide to abut against.

5. The centrifugal immunohistochemical staining method according to claim 1, characterized in that: In step S1, the height of the support portion is set to 0.1-0.6 mm.

6. The centrifugal immunohistochemical staining method according to any one of claims 1 to 5, characterized in that: The centrifugal dyeing disc is located in the cylinder, the upper part of the cylinder is provided with the drip dyeing head, the drip dyeing assembly comprises a liquid supply pipeline, a liquid pump and a reaction liquid bottle, and the liquid pump pumps the corresponding reaction liquid into the drip dyeing head.

7. The centrifugal immunohistochemical staining method according to claim 6, characterized in that: The cylinder is also connected with a hot air blowing pipe to blow hot air into the cylinder to keep the cylinder at a certain temperature.

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