阅读说明：本技术 一种用于免疫荧光法的振动装置 (Vibration device for immunofluorescence method ) 是由 闫亚平 程静美 封雪 李科 于 2021-09-23 设计创作，主要内容包括：本发明公开了一种用于免疫荧光法的振动装置,属于医疗检测装置领域,该装置采用腔体、振动单元、降温装置和隔音棉,采用隔音棉降低实验过程中的噪音,降温装置降低因振动而产生的热量,避免振动产生的热量对样本造成影响；通电后振动单元形成一种振动源,通过振动带动腔体内孵育盒中的载玻片的振动,进而促进载玻片上固定的抗原/抗体表位的暴露,加速与孵育样本中的抗体/抗原结合,缩短孵育时间,提高实验效率。(The invention discloses a vibration device for an immunofluorescence method, which belongs to the field of medical detection devices, and adopts a cavity, a vibration unit, a cooling device and soundproof cotton, wherein the soundproof cotton is adopted to reduce noise in the experimental process, the cooling device reduces heat generated by vibration, and the heat generated by vibration is prevented from influencing a sample; the vibrating unit forms a vibration source after electrification, drives the vibration of the glass slide in the incubation box in the cavity through vibration, further promotes the exposure of the fixed antigen/antibody epitope on the glass slide, accelerates the combination with the antibody/antigen in the incubation sample, shortens the incubation time, and improves the experiment efficiency.)

1. A vibration device for immunofluorescence, comprising: the cavity is arranged on the vibration unit; the cavity comprises a vibration cavity bottom plate (2) and a vibration cavity (1) arranged above the vibration cavity bottom plate and used for containing the incubation box; the vibration unit comprises a motor bottom plate (5) and a plurality of motors (4); the motors (4) are densely fixed on the motor bottom plate (5).

2. The vibration device for immunofluorescence according to claim 1, wherein, the motor bottom plate (5) is further provided with a cooling device (3).

3. The vibrating device for immunofluorescence according to claim 2, wherein, the cooling device (3) is a fan or a semiconductor cooling plate.

4. A vibration device for immunofluorescence according to claim 3, wherein, the cooling device (3) is arranged above or below the motor base plate (5) according to different types.

5. The vibration device for immunofluorescence according to claim 1, wherein there is further provided soundproof cotton (6) under the motor base plate (5).

6. The vibration device for immunofluorescence according to claim 1, wherein, the vibration chamber bottom plate (2) is made of rubber.

7. A vibration device for immunofluorescence according to claim 1, wherein, the circuit connection of several motors (4) adopts parallel connection.

8. A vibration device for immunofluorescence according to claim 1, wherein, the motor (4) is a flat rotor motor or a linear motor.

9. The vibration device for immunofluorescence according to claim 8, wherein, when the motor (4) is a linear motor, the vibration frequency is 100-299 HZ.

10. The vibration device for immunofluorescence according to claim 8, wherein, when the motor (4) is a flat rotor motor, the rated rotation speed is 8000 ± 2500rpm to 13000 ± 2500 rpm.

Technical Field

The invention belongs to the field of medical detection devices, and relates to a vibrating device for an immunofluorescence method.

Background

The immunofluorescence method is a technology for carrying out antigen positioning by marking an antibody with a fluorescent substance, and is characterized by strong specificity, obvious comparison of signal intensity of positive samples and negative samples, capability of accurately judging tissue or intracellular fluorescence distribution through microscope observation and becoming a standard technology for autoantibody diagnosis. Although the immunofluorescence method is widely applied to the detection process of the autoantibody, the incubation time of the primary antibody and the secondary antibody is 30 min-1 h or even longer in the existing immunofluorescence method detection, the whole detection process consumes long time, and the detection efficiency is low.

At present, the application of microwave technology to immunofluorescence is reported in documents, which can effectively shorten the time of the whole process, but the method is not widely applied, and the operation process is troublesome to some extent; there is a fear that the heat generated from the microwave during use affects the sample. It is reported in the literature that vibration enhances the immunofluorescence signal, and the shaking table in the prior art can generate shaking to help mix the sample, but the following problems can occur: the sample incubated on the climbing sheet flows out of the hole, so that no sample incubation and no result detection are caused; the liquid sample incubated on the creeper has a similar vortex phenomenon due to the shaking of the shaking table, so that the incubation of the sample is not uniform, and detection signals are inconsistent; a small amount of the sample incubated on the slide can flow out of the hole due to shaking of the shaking table, thereby causing cross contamination.

Disclosure of Invention

In order to overcome the disadvantages of the prior art, the present invention provides 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.

In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:

the invention discloses a vibration device for immunofluorescence, which comprises: the cavity is arranged on the vibration unit;

the cavity comprises a vibration cavity and a vibration cavity bottom plate; the vibration cavity is arranged on the vibration cavity bottom plate, and the incubation box is placed in the vibration cavity; the cavity comprises a vibration cavity bottom plate and a vibration cavity arranged above the vibration cavity bottom plate and used for containing the incubation box; the vibration unit comprises a motor bottom plate and a plurality of motors; the motors are densely fixed on the motor bottom plate.

Preferably, a cooling device is further arranged on the motor bottom plate.

Further preferably, the cooling device is a fan or a semiconductor cooling plate.

It is further preferred that the cooling devices are distributed above or below the motor floor depending on their shape.

Preferably, soundproof cotton is further disposed under the motor base plate.

Preferably, the bottom plate of the vibration cavity is made of rubber.

Preferably, the circuit connection of the plurality of motors is in parallel.

Preferably, the motor is a flat rotor motor or a linear motor.

More preferably, when the motor is a linear motor, the vibration frequency is 100 to 299 HZ.

Further preferably, when the motor is a flat rotor motor, the rated rotating speed is 8000 +/-2500 rpm-13000 +/-2500 rpm.

Compared with the prior art, the invention has the following beneficial effects:

the invention discloses a vibration device for an immunofluorescence method; the vibration unit drives the cavity to vibrate to promote the combination of the antigen and the antibody on the immunofluorescence plate in the cavity, and the vibration unit comprises a motor bottom plate and a plurality of motors; the motors are densely fixed on the motor bottom plate and generate irregular vibration after being electrified, the cavity comprises a vibration cavity and a vibration cavity bottom plate, the vibration cavity is arranged on the vibration cavity bottom plate, the irregular vibration of the motors drives the vibration cavity bottom plate and the vibration cavity to vibrate, and therefore the situation that in the vibration process, samples incubated on the immunofluorescence sheet with overlarge amplitude flow out of holes to cause no sample incubation of the climbing sheet and no result in detection is avoided; the phenomenon that the liquid sample incubated on the immunofluorescence plate is similar to 'vortex' due to vibration is avoided, so that the sample incubation is not uniform, and detection signals are not uniform; or a small amount of the sample incubated on the immunofluorescent sheet may flow out of the well due to vibration, causing cross contamination. The device is used for carrying out experiments, the experiment time of the immunofluorescence method can be shortened to 20-30 min from 1-2 h or even longer, and the experiment efficiency is effectively improved.

Furthermore, the inside heat sink that still is provided with of vibration unit, the heat that the heat sink can reduce to produce because of the motor vibration, avoids the heat that the vibration produced to cause the influence to the sample.

Furthermore, the fan or the semiconductor refrigerating sheet is used as a cooling device to reduce the heat generated by the vibration of the motor, so that the motor which generates the vibration in the vibration source can maintain the temperature of 18-25 ℃ in the working range, and the effect of detecting the antibody in the sample to be detected by the antigen fixed on the climbing sheet is not influenced.

Further, the vibration unit is arranged on the soundproof cotton, and the soundproof cotton can reduce noise in the experimental process.

Further, the vibration chamber bottom plate adopts the rubber material to make, and at the vibration in-process, the slide glass of placing in the vibration intracavity does not take place the position and removes, consequently, the sample of hatching on the cell climbing piece of pasting on the slide glass also can not flow out from the hole, the effectual combination of waiting to examine sample and cell climbing piece of having guaranteed.

Further, a plurality of motors are connected in parallel, so that the damage of the motors accounting for lower proportion of the total number of the motors 1/100, 2/100, 3/100, 4/100, 5/100 and the like does not influence the vibration effect of the device.

Furthermore, the motor on the motor bottom plate is a flat rotor motor or a linear motor, the two motors are densely distributed on the motor bottom plate, and irregular vibration is generated after electrification, so that the vibration effect required by the device is favorably generated.

Further, the vibration frequency of linear motor be 100 ~ 299HZ, linear motor produces the vibration according to certain frequency after the circular telegram, is favorable to forming a vibration source, and the vibration that this vibration source produced can make the liquid of incubating on the cell slide fully mix and matrix (cell/tissue) on the slide maintain good stationary state on the one hand, and on the other hand, this vibration source can not lead to incubating the liquid of on the slide and flow out from this sample hole and cause the slide not have the sample to incubate or cause other sample hole pollutions.

Furthermore, the rated rotation speed of the flat rotor motor is 8000 +/-2500 rpm-13000 +/-2500 rpm, and the motor rotates in any direction after being electrified, so that a disordered vibration source is favorably formed, on one hand, the vibration generated by the vibration source can enable liquid incubated on the cell climbing sheet to be fully mixed and a matrix (cells/tissues) on the climbing sheet to maintain a good fixed state, and on the other hand, the vibration source can not cause the liquid incubated on the climbing sheet to flow out of the sample hole to cause no sample incubation of the climbing sheet or cause the pollution of other sample holes.

Drawings

FIG. 1 is a schematic structural view of the present invention;

FIG. 2 is a schematic view of a flat motor base, a motor and a heat dissipation device according to the present invention;

FIG. 3 is a schematic diagram showing the effect of vibration incubation in an oscillator applied to immunofluorescence assay with the flat rotor motor of the present invention on anti-Hu antibody signal on CBA and anti-AQP 4 antibody signal on TBA; wherein (a) and (b) are graphs of experimental results of example 1, (c) and (d) are graphs of experimental results of comparative example 1, and (e) and (f) are graphs of experimental results of comparative example 2;

FIG. 4 is a schematic representation of the effect of inventive internal shaking incubation on anti-Tr antibody signal on CBA and anti-AQP 4 antibody signal on TBA assay, consisting of inventive linear motors; wherein (a) and (b) are graphs of experimental results of example 2, (c) and (d) are graphs of experimental results of comparative example 3, and (e) and (f) are graphs of experimental results of comparative example 4.

Wherein: 1-a vibration cavity; 2-vibrating chamber floor; 3-semiconductor refrigerating sheet; 4-a motor; 5-motor base plate; 6-soundproof cotton.

Detailed Description

In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

The invention is described in further detail below with reference to the accompanying drawings:

the invention aims to provide a vibrating device for an immunofluorescence method, which can form a vibration source after being electrified, and further promotes the exposure of fixed antigen/antibody epitopes on a glass slide by driving the vibration of the glass slide in an incubation box through vibration, thereby accelerating the combination with the antibody/antigen in an incubation sample, shortening the incubation time and improving the experimental efficiency.

Referring to fig. 1 and 2, the device comprises: a vibration chamber 1; a vibration cavity bottom plate 2; a cooling device 3; a motor 4; a motor base plate 5; 6, soundproof cotton; the vibration cavity 1 is arranged on a vibration cavity bottom plate 2, the motors 4 are connected in parallel and fixed on a motor bottom plate 5, the vibration cavity bottom plate 2 is arranged above the motors 4, the motor bottom plate 5 is arranged on soundproof cotton 6, and the cooling device 3 is arranged above the motor bottom plate 5;

the vibration cavity 1 can be used for placing an incubation box; the bottom plate 2 of the vibration cavity is a rubber pad; the motor 4 connected in parallel is fixed on the motor bottom plate 5, and the vibration of the motor 4 promotes the combination of the antigen and the antibody on the immunofluorescence sheet in the incubation box of the vibration cavity 1; the cooling device 3 can reduce the heat generated by the vibration of the motor, so that the motor generating the vibration in the vibration source 2 can maintain the temperature in the working range. The soundproof cotton 6 can reduce the noise in the experimental process; further, the vibration unit includes a motor base plate 5 and a number of motors 4, and generates a vibration source by vibration of the flat motor or the linear motor; the certain number can be connected according to actual requirements, and the number range is more than or equal to 6;

furthermore, the rotation direction of the motor 4 is any direction; the rated rotating speed of the motor 4 is 8000 plus or minus 2500rpm, 9000 plus or minus 2500rpm, 10000 plus or minus 3000rpm, 11000 plus or minus 2500rpm, 12000 plus or minus 2500rpm, 13000 plus or minus 2500 rpm; the vibration frequency of the motor 4 is 100 to 200HZ, 200 to 299 HZ.

Further, the motor 4 may be a rectangular parallelepiped linear motor, and may also be a flat rotor motor; generating a vibration source by vibration of a motor;

the cooling device 3 comprises the existing small cooling equipment, including a fan, a semiconductor refrigeration piece and the like, and can be distributed above or below the motor bottom plate according to the shape of the cooling device;

by adopting the device, 60 motors 4 rotating in any direction are welded on the motor bottom plate 5, the diameter of each motor 4 is 10mm, the thickness of each motor 4 is 2.7mm, the rotating speed is 11000 +/-2500 rpm, the using voltage range is 2.0-4.0V DC, and the motors 4 are connected in parallel; the cooling device 3 adopts a semiconductor refrigeration piece;

example 1 applying the above-described vibration apparatus for immunofluorescence assay to a cell-based immunofluorescence assay, the cell slide being a cell slide overexpressing the Hu antigen, the primary antibody being a positive sample against the Hu antigen, specifically includes the following steps:

(1) washing the slide with PBS;

(2) an anti-vibration incubation: incubating the positive sample for 15 min;

(3) vibration washing: wash 3 times with PBST;

(4) and (3) performing anti-vibration incubation: incubating for 15min for secondary antibody;

(5) vibration washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

The vibration device for the immunofluorescence assay is applied to the immunofluorescence assay based on tissues, the tissue slide is a mouse hypothalamus, and a primary antibody is a positive sample of an anti-AQP 4 antigen, and the vibration device specifically comprises the following steps:

(1) an anti-vibration incubation: incubating the positive sample for 15 min;

(2) washing: wash 3 times with PBST;

(3) and (3) performing anti-vibration incubation: incubating for 15min for secondary antibody;

(4) washing: wash 3 times with PBST;

(5) and observing the result under a microscope.

Comparative example 1, in contrast to the above device, incubation times with primary and secondary antibodies were different and the experiment was carried out without vibration throughout the experiment:

(1) washing the slide with PBS (tissue slide does not require this step, starting directly from step (2));

(2) primary antibody incubation: incubating the positive sample for a first-antibody time of 60 min;

(3) washing: wash 3 times with PBST;

(4) and (3) secondary antibody incubation: incubating for 40min for the second antibody;

(5) washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

Comparative example 2, in contrast to the above device, no vibration was applied during the whole experiment:

(1) washing the slide with PBS (tissue slide does not require this step, starting directly from step (2));

(2) primary antibody incubation: incubating the positive sample for 15 min;

(3) washing: wash 3 times with PBST;

(4) and (3) secondary antibody incubation: incubating for 15min for secondary antibody;

(5) washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

Comparative results referring to fig. 3, the effect of the vibration device for immunofluorescence consisting of a flat motor on the positive signal in the CBA method and TBA method is shown schematically. In FIG. 3, (a) is CBA method Hu⁺(b) is TBA method AQP4⁺The positive signals were greatly influenced by the CBA method and TBA method using a vibrating device for immunofluorescence, and the positive signals of example 1 were compared with those of (c)(d) The incubation times with the primary and secondary antibodies are shown to be different and the positive signals are more evident in comparative examples 1 and (e) and (f) where the experiment was carried out without vibration throughout the experiment and comparative example 2 where there was no vibration throughout the experiment.

Embodiment 2, adopt the vibrating device for immunofluorescence of fig. 1, the motor is a linear motor, wherein 60 motors 4 with 205Hz amplitude are distributed on the motor bottom plate 5, the length, width and height of the motor 4 are respectively 12mm, 4mm and 3mm, the rated input voltage is 2.0 ± 0.05Vrms AC, the connection mode of each motor 4 is parallel, the cooling device 3 adopts semiconductor refrigeration piece; the vibration device for the immunofluorescence method is applied to the cell-based immunofluorescence method, the cell slide is a cell slide over-expressing Tr antigen, and the primary antibody is a positive sample resisting the Tr antigen, and the method specifically comprises the following steps:

(1) washing the slide with PBS;

(2) an anti-vibration incubation: incubating the positive sample for 15 min;

(3) vibration washing: wash 3 times with PBST;

(4) and (3) performing anti-vibration incubation: incubating for 15min for secondary antibody;

(5) vibration washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

The vibration device for the immunofluorescence assay is applied to the immunofluorescence assay based on tissues, the tissue slide is a mouse hypothalamus, and a primary antibody is a positive sample of an anti-AQP 4 antigen, and the vibration device specifically comprises the following steps:

(1) an anti-vibration incubation: incubating the positive sample for 15 min;

(2) washing: wash 3 times with PBST;

(3) and (3) performing anti-vibration incubation: incubating for 15min for secondary antibody;

(4) washing: wash 3 times with PBST;

(5) and observing the result under a microscope.

Comparative example 3, in comparison to the above device, incubation times with primary and secondary antibodies were different and no vibration was applied to the comparative experiment throughout the experiment:

(1) washing the slide with PBS (tissue slide does not require this step, starting directly from step (2));

(2) an anti-vibration incubation: incubating the positive sample for a first-antibody time of 60 min;

(3) vibration washing: wash 3 times with PBST;

(4) and (3) performing anti-vibration incubation: incubating for 40min for the second antibody;

(5) vibration washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

Comparative example 4, in comparison to the above apparatus, a vibration-free comparative experiment was carried out throughout the experiment:

(1) washing the slide with PBS (tissue slide does not require this step, starting directly from step (2));

(2) primary antibody incubation: incubating the positive sample for 15 min;

(3) washing: wash 3 times with PBST;

(4) and (3) secondary antibody incubation: incubating for 15min for secondary antibody;

(5) washing: wash 3 times with PBST;

(6) and observing the result under a microscope.

Comparative results referring to fig. 4, the effect of the oscillator composed of linear motor applied to immunofluorescence assay on positive signals in CBA and TBA assay is shown schematically. As shown in the figure, (a) in FIG. 4 is CBA method Tr⁺(b) is TBA method AQP4⁺Example 2 the effect of the oscillator applied to the immunofluorescence method on the positive signals in the CBA method and TBA method was much larger than the difference in incubation time with the primary antibody and the secondary antibody shown in (c) and (d), and comparative example 3 where the experiment was carried out without vibration throughout the experiment, and comparative example 4 where the experiment was carried out without vibration throughout the experiment shown in (e) and (f).

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.