A vibration device for immunofluorescence

Abstract

The application discloses a kind of vibration device for immunofluorescence method, belong to medical detection device field, the device adopts cavity, vibration unit, cooling device and sound insulation cotton, sound insulation cotton is used to reduce noise in experimental process, cooling device reduces the heat generated due to vibration, avoid the heat generated by vibration to cause the influence to sample;After energization, vibration unit forms a kind of vibration source, the vibration of slide in incubation box in cavity is driven by vibration, and then promote the exposure of fixed antigen / antibody epitope on slide, accelerate and antibody / antigen in incubation sample are combined, shorten incubation time, improve experimental efficiency.

Description

Technical Field

[0001] This invention belongs to the field of medical testing devices and relates to a vibration device for immunofluorescence assay. Background Technology

[0002] Immunofluorescence is a technique for antigen localization using fluorescently labeled antibodies. This method is characterized by high specificity, a clear contrast in signal intensity between positive and negative samples, and precise microscopic observation of intracellular fluorescence distribution. It has become the standard technique for autoantibody diagnosis. Although immunofluorescence is widely used in autoantibody detection, current immunofluorescence assays require incubation times of 30 minutes to 1 hour or even longer for both primary and secondary antibodies, resulting in a time-consuming process and low detection efficiency.

[0003] Currently, existing literature reports the application of microwave technology to immunofluorescence, which can effectively shorten the entire process time. However, this method has not been widely adopted, possibly due to the relatively cumbersome operation and concerns about the heat generated by microwaves affecting the samples. Some literature reports that vibration enhances the immunofluorescence signal. While existing shakers generate vibration, which helps mix the samples, the following problems may occur: samples incubated on the slide may flow out of the wells, resulting in no sample incubation and no detection results; liquid samples incubated on the slide may exhibit a "vortex" phenomenon due to the shaker's movement, leading to uneven sample incubation and inconsistent detection signals; and a small amount of sample incubated on the slide may flow out of the wells due to the shaker's movement, causing cross-contamination. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the present invention aims to provide a vibration device for immunofluorescence assay, so as to solve the problems of long detection time, low working efficiency and easy cross-contamination in the prior art.

[0005] To achieve the above objectives, the present invention employs the following technical solution:

[0006] The present invention discloses a vibration device for immunofluorescence assay, comprising: a cavity and a vibration unit, wherein the cavity is disposed on the vibration unit;

[0007] The cavity includes a vibration chamber and a vibration chamber base plate; the vibration chamber is disposed on the vibration chamber base plate, and an incubation box is placed inside the vibration chamber; the cavity includes a vibration chamber base plate and a vibration chamber disposed above it for holding the incubation box; the vibration unit includes a motor base plate and several motors; the several motors are densely fixed on the motor base plate.

[0008] Preferably, a cooling device is also provided on the motor base plate.

[0009] More preferably, the cooling device is a fan or a semiconductor cooling chip.

[0010] More preferably, the cooling device is distributed above or below the motor base plate according to its shape.

[0011] Preferably, sound insulation cotton is also provided below the motor base plate.

[0012] Preferably, the bottom plate of the vibration chamber is made of rubber.

[0013] Preferably, the circuits of the plurality of motors are connected in parallel.

[0014] Preferably, the motor is a flat rotor motor or a linear motor.

[0015] More preferably, when the motor is a linear motor, the vibration frequency is 100–299 Hz.

[0016] More preferably, when the motor is a flat rotor motor, the rated speed is 8000±2500rpm~13000±2500rpm.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] This invention discloses a vibration device for immunofluorescence assays. A vibration unit drives a cavity to vibrate, promoting the binding of antigens and antibodies on the immunofluorescence slide within the cavity. The vibration unit includes a motor base plate and several motors. These motors are densely arranged and fixed on the motor base plate, generating irregular vibrations upon energization. The cavity includes a vibration chamber and a vibration chamber base plate. The vibration chamber is positioned on the vibration chamber base plate. The irregular vibrations of the motors drive the vibration chamber base plate and the vibration chamber to vibrate, preventing excessive amplitude vibrations that could cause the sample incubated on the immunofluorescence slide to flow out of the wells, resulting in no sample incubation and no detection results. It also prevents the liquid sample incubated on the immunofluorescence slide from developing a "vortex" phenomenon due to vibration, leading to uneven sample incubation and inconsistent detection signals; or prevents a small amount of sample incubated on the immunofluorescence slide from flowing out of the wells due to vibration, causing cross-contamination. Using this device, the experimental time for immunofluorescence assays can be shortened from 1-2 hours or even longer to 20-30 minutes, effectively improving experimental efficiency.

[0019] Furthermore, a cooling device is installed inside the vibration unit to reduce the heat generated by the motor vibration and prevent the heat generated by the vibration from affecting the sample.

[0020] Furthermore, fans or semiconductor cooling devices are used as cooling devices to reduce the heat generated by motor vibration, so that the motor that generates vibration in the vibration source maintains the temperature within the working range of 18℃~25℃, while not affecting the effect of antigen detection on the sample to be tested by the antigen fixed on the climbing plate.

[0021] Furthermore, the vibration unit is placed on sound-absorbing cotton, which can reduce noise during the experiment.

[0022] Furthermore, the bottom plate of the vibration chamber is made of rubber. During vibration, the glass slide placed inside the vibration chamber does not move. Therefore, the sample incubated on the cell spreader attached to the glass slide will not flow out of the well, effectively ensuring the binding of the sample to be tested with the cell spreader.

[0023] Furthermore, several motors are connected in parallel, so that if a motor representing a relatively low proportion (1 / 100, 2 / 100, 3 / 100, 4 / 100, 5 / 100, etc.) fails, the vibration effect of the device will not be affected.

[0024] Furthermore, the motors on the motor base plate are either flat rotor motors or linear motors. These two types of motors are densely distributed on the motor base plate, and when energized, they generate irregular vibrations, which is beneficial to producing the vibration effect required by this device.

[0025] Furthermore, the linear motor has a vibration frequency of 100-299 Hz. When powered on, the linear motor vibrates at a certain frequency, which helps to form a vibration source. On the one hand, the vibration generated by this vibration source will make the liquid incubated on the cell slide fully mixed and the matrix (cells / tissues) on the slide maintain a good fixed state. On the other hand, this vibration source will not cause the liquid incubated on the slide to flow out of the sample well, resulting in no sample incubation on the slide or contamination of other sample wells.

[0026] Furthermore, the rated speed of the flat rotor motor is 8000±2500rpm~13000±2500rpm. After being powered on, the motor rotates in any direction, which is conducive to forming a disordered vibration source. On the one hand, the vibration generated by this vibration source will make the liquid incubated on the cell slide fully mixed and the matrix (cells / tissues) on the slide maintain a good fixed state. On the other hand, this vibration source will not cause the liquid incubated on the slide to flow out of the sample well, resulting in no sample incubation on the slide or contamination of other sample wells. Attached Figure Description

[0027] Figure 1 This is a schematic diagram of the structure of the present invention;

[0028] Figure 2 A schematic diagram of the invention's flat motor base plate, motor, and heat dissipation device;

[0029] Figure 3 The diagram shows the effect of vibration incubation in an oscillator constructed from the flat rotor motor of the invention on the anti-Hu antibody signal in the CBA method and the anti-AQP4 antibody signal in the TBA method; where (a) and (b) are experimental results of Example 1, (c) and (d) are experimental results of Comparative Example 1, and (e) and (f) are experimental results of Comparative Example 2.

[0030] Figure 4 The diagram shows the effect of the internal vibration incubation of the linear motor of the invention on the anti-Tr antibody signal in the CBA method and the anti-AQP4 antibody signal in the TBA method; where (a) and (b) are the experimental results of Example 2, (c) and (d) are the experimental results of Comparative Example 3, and (e) and (f) are the experimental results of Comparative Example 4.

[0031] Wherein: 1-Vibration chamber; 2-Vibration chamber base plate; 3-Semiconductor cooling chip; 4-Motor; 5-Motor base plate; 6-Sound insulation cotton. Detailed Implementation

[0032] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0033] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0034] The present invention will now be described in further detail with reference to the accompanying drawings:

[0035] The purpose of this invention is to provide a vibration device for immunofluorescence assay. When the device is powered on, it forms a vibration source, which drives the vibration of the glass slide in the incubation chamber, thereby promoting the exposure of the antigen / antibody epitopes fixed on the glass slide, accelerating the binding with antibodies / antigens in the incubation sample, shortening the incubation time, and improving experimental efficiency.

[0036] See Figure 1 , Figure 2 The device includes: a vibration chamber 1; a vibration chamber base plate 2; a cooling device 3; a motor 4; a motor base plate 5; and sound insulation cotton 6. The vibration chamber 1 is set on the vibration chamber base plate 2, the motors 4 are connected in parallel and fixed on the motor base plate 5, the vibration chamber base plate 2 is set above the motors 4, the motor base plate 5 is set on the sound insulation cotton 6, and the cooling device 3 is set above the motor base plate 5.

[0037] The vibration chamber 1 can hold an incubation box; the bottom plate 2 of the vibration chamber is a rubber pad; motors 4 connected in parallel are fixed on the motor base plate 5, and the vibration of the motors 4 promotes the binding of antigens and antibodies on the immunofluorescence slides in the incubation box of the vibration chamber 1; the cooling device 3 can reduce the heat generated by the motor vibration, so that the motors vibrating in the vibration source 2 can maintain the temperature within the working range. The sound insulation cotton 6 can reduce the noise during the experiment; furthermore, the vibration unit includes a motor base plate 5 and a certain number of motors 4, and the vibration source is generated by the vibration of flat motors or linear motors; the certain number can be connected according to actual needs, and the number range is ≥ 6;

[0038] Furthermore, the rotation direction of the motor 4 is arbitrary; the rated speed of the motor 4 is 8000±2500rpm, 9000±2500rpm, 10000±3000rpm, 11000±2500rpm, 12000±2500rpm, and 13000±2500rpm; the vibration frequency of the motor 4 is 100~200HZ and 200~299HZ.

[0039] Furthermore, the motor 4 can be a cuboid linear motor or a flat rotor motor; the vibration of the motor generates a vibration source.

[0040] The cooling device 3 includes existing small cooling devices, such as fans and semiconductor cooling chips, which can be distributed above or below the motor base plate depending on the shape of the cooling device.

[0041] The above-mentioned device has 60 motors 4 that rotate in any direction welded on the motor base plate 5. The motors 4 have a diameter of 10 mm, a thickness of 2.7 mm, a speed of 11000±2500 rpm, and a voltage range of 2.0~4.0V DC. The motors 4 are connected in parallel. The cooling device 3 uses a semiconductor refrigeration chip.

[0042] Example 1 applies the vibration device described above for immunofluorescence to cell-based immunofluorescence, where the cell slide is a cell slide overexpressing Hu antigen, and the primary antibody is a positive sample against Hu antigen. The specific steps include:

[0043] (1) Wash the slides with PBS;

[0044] (2) Primary antibody vibration incubation: Incubate positive samples with primary antibody for 15 min;

[0045] (3) Vibration washing: Wash 3 times with PBST;

[0046] (4) Secondary antibody vibration incubation: Secondary antibody incubation time is 15 min;

[0047] (5) Vibration washing: Wash 3 times with PBST;

[0048] (6) Observation results under a microscope.

[0049] The vibration device described above for immunofluorescence is applied to a tissue-based immunofluorescence assay, where the tissue slide is mouse hypothalamus and the primary antibody is anti-AQP4 antigen. The specific steps include:

[0050] (1) Primary antibody vibration incubation: Incubate positive samples with primary antibody for 15 min;

[0051] (2) Washing: Wash 3 times with PBST;

[0052] (3) Secondary antibody vibration incubation: Secondary antibody incubation time is 15 min;

[0053] (4) Washing: Wash 3 times with PBST;

[0054] (5) Observation results under a microscope.

[0055] Comparative Example 1, compared to the above apparatus, uses different incubation times for the primary and secondary antibodies, and the entire experiment is conducted without vibration:

[0056] (1) Wash the slide with PBS (this step is not required for tissue slides; proceed directly from step (2));

[0057] (2) Primary antibody incubation: Incubate positive samples with primary antibody for 60 min;

[0058] (3) Washing: Wash 3 times with PBST;

[0059] (4) Secondary antibody incubation: Secondary antibody incubation time is 40 min;

[0060] (5) Washing: Wash 3 times with PBST;

[0061] (6) Observation results under a microscope.

[0062] Comparative Example 2, compared to the above apparatus, uses a vibration-free experimental procedure throughout the entire process:

[0063] (1) Wash the slide with PBS (this step is not required for tissue slides; proceed directly from step (2));

[0064] (2) Primary antibody incubation: Incubate positive samples with primary antibody for 15 min;

[0065] (3) Washing: Wash 3 times with PBST;

[0066] (4) Secondary antibody incubation: Secondary antibody incubation time is 15 min;

[0067] (5) Washing: Wash 3 times with PBST;

[0068] (6) Observation results under a microscope.

[0069] See comparison results Figure 3 A schematic diagram illustrating the effect of a vibrating device consisting of a flat motor, used in immunofluorescence assays, on positive signals in the CBA and TBA methods. Figure 3 In the middle, (a) is the CBA law Hu + (b) is the TBA method AQP4 + The vibration device used in immunofluorescence assays has a significant impact on the positive signal in both CBA and TBA methods. Compared with the positive signal in Example 1, the positive signal in Comparative Example 1 and Comparative Example 2, which was conducted without vibration throughout the entire experimental process, showed a more obvious positive signal. (c) and (d) showed different incubation times for the primary and secondary antibodies.

[0070] Example 2, using Figure 1 The vibration device used in immunofluorescence assay employs a linear motor. Sixty motors 4, each with an amplitude of 205 Hz, are distributed on the motor base plate 5. The length, width, and height of each motor 4 are 12 mm, 4 mm, and 3 mm, respectively. The rated input voltage is 2.0 ± 0.05 Vrms AC. The motors 4 are connected in parallel. The cooling device 3 uses a semiconductor thermoelectric cooler. This vibration device for immunofluorescence assay is applied to cell-based immunofluorescence assays. The cell slide is an overexpression of Tr antigen, and the primary antibody is a positive sample against the Tr antigen. The specific steps include:

[0071] (1) Wash the slides with PBS;

[0072] (2) Primary antibody vibration incubation: Incubate positive samples with primary antibody for 15 min;

[0073] (3) Vibration washing: Wash 3 times with PBST;

[0074] (4) Secondary antibody vibration incubation: Secondary antibody incubation time is 15 min;

[0075] (5) Vibration washing: Wash 3 times with PBST;

[0076] (6) Observation results under a microscope.

[0077] The vibration device described above for immunofluorescence is applied to a tissue-based immunofluorescence assay, where the tissue slide is mouse hypothalamus and the primary antibody is anti-AQP4 antigen. The specific steps include:

[0078] (1) Primary antibody vibration incubation: Incubate positive samples with primary antibody for 15 min;

[0079] (2) Washing: Wash 3 times with PBST;

[0080] (3) Secondary antibody vibration incubation: Secondary antibody incubation time is 15 min;

[0081] (4) Washing: Wash 3 times with PBST;

[0082] (5) Observation results under a microscope.

[0083] Comparative Example 3, compared to the above apparatus, uses different incubation times for the primary and secondary antibodies, and is a comparative experiment conducted without vibration throughout the entire experimental process:

[0084] (1) Wash the slide with PBS (this step is not required for tissue slides; proceed directly from step (2));

[0085] (2) Primary antibody vibration incubation: Incubate positive samples with primary antibody for 60 min;

[0086] (3) Vibration washing: Wash 3 times with PBST;

[0087] (4) Secondary antibody vibration incubation: Secondary antibody incubation time is 40 min;

[0088] (5) Vibration washing: Wash 3 times with PBST;

[0089] (6) Observation results under a microscope.

[0090] Comparative Example 4: In contrast to the above apparatus, a control experiment was conducted with no vibration throughout the entire experimental process.

[0091] (1) Wash the slide with PBS (this step is not required for tissue slides; proceed directly from step (2));

[0092] (2) Primary antibody incubation: Incubate positive samples with primary antibody for 15 min;

[0093] (3) Washing: Wash 3 times with PBST;

[0094] (4) Secondary antibody incubation: Secondary antibody incubation time is 15 min;

[0095] (5) Washing: Wash 3 times with PBST;

[0096] (6) Observation results under a microscope.

[0097] See comparison results Figure 4 This is a schematic diagram illustrating the effect of a linear motor-based oscillator on the positive signal in CBA and TBA methods for immunofluorescence. (See figure.) Figure 4 (a) is the CBA method Tr + (b) is the TBA method AQP4 + Example 2 shows that the oscillator used in immunofluorescence assays has a much greater impact on the positive signal in CBA and TBA methods than Comparative Example 3, which uses different incubation times for primary and secondary antibodies and is conducted without vibration throughout the entire experimental process, and Comparative Example 4, which is conducted without vibration throughout the entire experimental process, as shown in (c) and (d).

[0098] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A vibration device for immunofluorescence, characterized in that, The application relates to a vibration incubator, which comprises the following parts: a cavity and a vibration unit, wherein the cavity is arranged on the vibration unit; the cavity comprises a vibration cavity bottom plate (2) and a vibration cavity (1) for containing an incubation box arranged above the vibration cavity bottom plate (2); the vibration unit comprises a motor bottom plate (5) and a plurality of motors (4); the plurality of motors (4) are fixed on the motor bottom plate (5) in a dense manner; the number of the motors (4) is greater than or equal to 6, and the circuit connection of the plurality of motors (4) adopts parallel connection; the motor (4) is a flat rotor motor or a linear motor, and the motor (4) generates irregular vibration after being electrified; when the motor (4) is a linear motor, the vibration frequency is 100-299 HZ; when the motor (4) is a flat rotor motor, the rated rotating speed is 8000+ / -2500 rpm-13000+ / -2500 rpm; the vibration cavity bottom plate (2) is made of rubber.

2. A vibration device for immunofluorescence according to claim 1, characterized in that, A cooling device (3) is further arranged on the motor bottom plate (5).

3. A vibration device for immunofluorescence according to claim 2, characterized in that, The cooling device (3) is a fan or a semiconductor refrigerating sheet.

4. A vibration device for immunofluorescence according to claim 3, characterized in that, The cooling device (3) is arranged above or below the motor bottom plate (5) according to different types.

5. A vibration device for immunofluorescence according to claim 1, characterized in that, Soundproof cotton (6) is further arranged below the motor bottom plate (5).

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