CN210690354U

CN210690354U - Immunofluorescence analysis device

Info

Publication number: CN210690354U
Application number: CN201921062876.3U
Authority: CN
Inventors: 尹芝南; 付强强; 吴扬哲
Original assignee: Jinan University
Current assignee: Jinan University; University of Jinan
Priority date: 2019-07-08
Filing date: 2019-07-08
Publication date: 2020-06-05
Anticipated expiration: 2029-07-08

Abstract

The utility model provides an immunofluorescence analysis device, immunofluorescence analysis device includes the black filter membrane, immunofluorescence analysis device's application method is: intercepting an object to be detected dyed by a fluorescence labeling antibody by adopting a black filter membrane, filtering the fluorescence labeling antibody which is not combined with the object to be detected, and analyzing the intercepted matters on the surface of the black filter membrane by adopting a fluorescence detection device to realize reading and counting of fluorescence points; the immunofluorescence analysis device is simple in structure, low in production cost, convenient to use and good in detection effect; the immunofluorescence analysis device of the utility model can be widely used, is not only suitable for laboratories of large hospitals and major, but also can be widely used in basic medical institutions.

Description

Immunofluorescence analysis device

Technical Field

The utility model relates to an immunofluorescence detects the field, especially relates to an immunofluorescence assay device.

Background

Cell (including human body cell, fungus, bacteria, etc.) counting has important application in clinical diagnosis, food safety, environmental pollution monitoring, etc.

The immunofluorescence method is widely applied to cell counting, and the main principle is that specific protein markers possibly exist on the surface of each cell, and fluorescence imaging of specific cells can be realized through antibody staining marked by fluorescent dyes corresponding to the markers, so that cell counting can be realized through counting the number of fluorescent points.

The existing immunofluorescence counting method mainly comprises the following steps:

step 1, pretreating a sample to be detected;

step 2, mixing a sample to be detected with the fluorescence labeling antibody to obtain a mixed system; the sample to be detected contains an object to be detected (such as an object to be detected and the like) capable of being combined with the fluorescent labeled antibody;

step 3, removing the fluorescence labeled antibody which is not combined with the object to be detected in the mixed system to obtain a sample to be analyzed;

and 4, counting the objects to be detected in the sample to be analyzed by utilizing the characteristic that the fluorescence labeled antibody emits fluorescence when being irradiated by light with a specific wavelength.

The above immunofluorescence counting method has the following problems:

(1) some substances (such as red blood cells in blood) which have obvious interference on fluorescent signals exist in a sample to be detected, so that the interference of the interfering substances on a detection result needs to be reduced through multi-step operation in the pretreatment process of the sample to be detected, and the operation process is complicated.

(2) After staining the cells with fluorochrome-labeled antibodies, centrifugation is required to remove the antibodies that are not bound to the cells, thereby eliminating background interference. This process requires multiple washing operations, requires the use of a centrifuge and a washing buffer, and is complicated and costly.

(3) The counting of step 4 is usually implemented by using a flow cytometer or other devices, which are expensive, require professional personnel to operate, require regular maintenance, and have high operation cost.

Since the existing immunofluorescence counting method has the above disadvantages, the immunofluorescence counting method is mainly applied to large hospitals or professional laboratories at present, and the immunofluorescence counting method cannot be widely applied to basic medical institutions (such as community health hospitals, clinics, families and the like) because the medical conditions of the basic medical institutions are poor, professionals with operation capability are lacked, and high equipment cost cannot be borne.

SUMMERY OF THE UTILITY MODEL

An object of the utility model is to provide an immunofluorescence analysis device, simple structure, low in production cost, convenient to use, detection effect is good.

In order to achieve the above object, the present invention provides an immunofluorescence analysis apparatus, comprising a black filter membrane.

In the first embodiment of the present invention, the immunofluorescence analysis apparatus further comprises a housing and a first water-absorbing material, the housing is provided with a first sample adding hole and a mounting groove which are sequentially communicated, the mounting groove is provided with a black filter membrane and a first water-absorbing material which are arranged in a stacked manner, wherein the black filter membrane is close to the first sample adding hole.

Optionally, the color of the shell is black, and the first water absorbing material is water absorbing paper.

In a second embodiment of the present invention, the immunofluorescence analysis apparatus further includes an upper cover and a lower casing which are mutually connected, the upper cover is provided with a second sample adding hole therein, the lower casing has an upper cover mating end and an air exhaust end which are arranged relatively, the black filter membrane is fixed between the upper cover and the upper cover mating end of the lower casing, a hollow structure is provided in the lower casing, and the hollow structure runs through the upper cover mating end and the air exhaust end.

Optionally, a protruding part is arranged at the matching end of the upper cover, the upper cover is connected with the protruding part in a matching manner, and the black filter membrane is fixed between the upper cover and the protruding part; the hollow structure comprises a first through hole which is arranged at the upper cover matching end of the lower shell and penetrates through the protruding part and a first groove which is arranged at the air exhaust end of the lower shell, and the bottom of the first groove is communicated with the first through hole.

Optionally, the upper cover is connected with the protruding portion in a clamping manner, and the color of the upper cover is black.

In a third embodiment of the present invention, the immunofluorescence analysis apparatus further comprises an upper housing, a fixed plug, a spring plate, and a base;

the upper shell is provided with a sample adding end and a base matching end which are arranged oppositely, the sample adding end is provided with a second sample adding hole, the base matching end is provided with a second groove, the second sample adding hole is communicated with the bottom of the second groove, and the fixed plug is fixed in the second groove;

the black filter membrane is arranged on the end face of one end, close to the second sampling hole, of the fixed plug;

a second through hole is formed in the fixing plug, and penetrates through one end, close to the second sampling hole, of the fixing plug and one end, close to the base, of the fixing plug;

the base comprises a bottom plate and a side plate connected with the bottom plate, an opening is formed in the bottom plate, the elastic sheet is borne on the bottom plate, and the upper shell is arranged on the elastic sheet and connected with the base; and a gap is arranged between one end of the fixed plug close to the base and the elastic sheet.

Optionally, the color of the upper shell is black, the fixing plug is fixed in the second groove in a clamping and bonding mode, and the distance between one end, close to the base, of the fixing plug and the elastic sheet is 2-5 mm.

Optionally, a second water-absorbing material is further fixed in the second groove, the second water-absorbing material is tightly attached to one side, close to the base, of the fixing plug, and a gap is formed between the second water-absorbing material and the elastic sheet.

Optionally, a third through hole corresponding to the second through hole is formed in the second water absorbing material.

The utility model has the advantages that:

the utility model provides an immunofluorescence analysis device, immunofluorescence analysis device includes the black filter membrane, immunofluorescence analysis device's application method is: intercepting an object to be detected dyed by a fluorescence labeling antibody by adopting a black filter membrane, filtering the fluorescence labeling antibody which is not combined with the object to be detected, and analyzing the intercepted matters on the surface of the black filter membrane by adopting a fluorescence detection device to realize reading and counting of fluorescence points; the utility model can obviously weaken the reflection effect of the filter membrane by setting the color of the filter membrane to be black, and avoid the situations of inaccurate reading and counting of fluorescent spots caused by the reflection of the filter membrane; the immunofluorescence analysis device is simple in structure, low in production cost, convenient to use and good in detection effect; the immunofluorescence analysis device of the utility model can be widely used, is not only suitable for laboratories of large hospitals and major, but also can be widely used in basic medical institutions.

In order to make the aforementioned and other objects, features and advantages of the present invention more comprehensible, preferred embodiments accompanied with figures are described in detail below.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention, and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1A is a schematic exploded view of a first embodiment of an immunofluorescence analysis apparatus according to the present invention;

FIG. 1B is a schematic cross-sectional view of a first embodiment of an immunofluorescence analysis apparatus according to the present invention;

FIG. 2A is a schematic exploded view of a second embodiment of the immunofluorescence analysis apparatus of the present invention;

FIG. 2B is a schematic cross-sectional view of a second embodiment of the immunofluorescence analysis apparatus of the present invention;

FIG. 3A is a schematic exploded view of a third embodiment of an immunofluorescence analysis apparatus according to the present invention;

FIG. 3B is a schematic cross-sectional view of a third embodiment of the immunofluorescence analysis apparatus of the present invention;

FIG. 4 is a schematic diagram of the immunofluorescence analysis method of the present invention.

Description of the main element symbols:

10. a housing; 11. a first well; 12. mounting grooves; 20. a black filter membrane; 31. a first water absorbent material; 41. an upper cover; 412. a second well; 20. a black filter membrane; 42. a lower housing; 421. a boss portion; 51. a first through hole; 61. a first groove; 71. an upper housing; 713. a third well; 714. a second groove; 715. a fixed plug; 716. a second through hole; 717. a cavity; 20. a black filter membrane; 72. a base; 721. a base plate; 723. an opening; 722. a side plate; 32. a second water absorbent material; 323. a third through hole; 80. an elastic sheet.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are exemplary only for the purpose of explaining the present invention, and should not be construed as limiting the present invention.

It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. In contrast, when an element is referred to as being "directly on" another element, there are no intervening elements present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," and "fixed" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically limited otherwise.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the templates herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Referring to fig. 1A to 3B, the present invention provides an immunofluorescence analysis apparatus, which includes a black filter 20.

The utility model discloses a black filter membrane 20 filters the sample to be analyzed that contains the object to be detected (such as cell and/or microorganism) that is dyed by the fluorescence labeling antibody, later adopts fluorescence detection device to detect the retention on black filter membrane 20 surface, and this fluorescence analysis method is simple and easy to operate, the result is accurate, the utility model discloses immunofluorescence analysis device's simple structure, low in production cost; additionally, the utility model discloses a set the filter membrane colour for black, can show the reflection of light effect that weakens the filter membrane, avoid because the filter membrane reflection of light leads to the fluorescence point to read and count the emergence of the inaccurate circumstances such as.

Specifically, the pore size of the filter pores in the black filter 20 is larger than the size of the fluorescently labeled antibody and smaller than the size of the object to be detected (e.g., cell and/or microorganism) stained with the fluorescently labeled antibody.

Referring to fig. 4, based on the above immunofluorescence analysis apparatus, the present invention provides an immunofluorescence analysis method, including:

providing a sample to be analyzed, which is a mixed system comprising a sample to be detected (such as blood, a bacterial sample, a secretion and the like) and a fluorescent labeled antibody, an immunofluorescence analysis device and the fluorescence detection device, wherein the sample to be detected contains an object to be detected (such as a cell and/or a microorganism);

adding a sample to be analyzed to the surface of the black filter membrane 20 for filtration, wherein liquid in the sample to be analyzed, a fluorescence labeled antibody which is not combined with an object to be detected and other tiny impurities can be filtered from the filter holes of the black filter membrane 20, and the object to be detected dyed by the fluorescence labeled antibody cannot pass through the filter holes due to large volume, so that the object to be detected can be trapped on the surface of the black filter membrane 20;

and reading and counting the retentate on the surface of the black filter membrane 20 by using a fluorescence detection device (such as a fluorescence microscope), and converting the number of the objects to be detected in the sample to be analyzed according to the amount of the sample added on the surface of the black filter membrane 20 and the total volume of the sample to be analyzed.

It is understood that the size of the object to be detected is usually in the micrometer (μm) level, and the size of the fluorescent labeled antibody is usually in the nanometer (nm) level, so that the fluorescent labeled antibody and other minute impurities which are not bound to the object to be detected can be smoothly filtered from the black filter 20 and the object to be detected stained by the fluorescent labeled antibody can be effectively trapped on the surface of the black filter 20 by properly designing the pore size of the filter pores on the black filter 20.

In some embodiments, the diameter of the object to be detected (cell or microorganism) is in the range of 2-12 μm, and the diameter of the fluorescent labeled antibody is in the range of 3-10nm, in which case, the pore size of the filter pores in the black filter 20 can be designed to be in the range of 1-10 μm, and the specific pore size is determined according to the actual size of the object to be detected, and the pore size can effectively ensure that the fluorescent labeled antibody which is not bound to the object to be detected is filtered from the black filter 20, and simultaneously ensure that the object to be detected which is stained by the fluorescent labeled antibody is effectively retained on the surface of the black filter 20.

Specifically, the material of the black filter 20 includes a substrate and a black pigment dispersed in the substrate.

Optionally, the substrate comprises one or more of polycarbonate, cellulose acetate, plant fiber, and the like.

Alternatively, the black pigment may be an inorganic black pigment (e.g., carbon black) or an organic black pigment.

Fig. 1A and fig. 1B are respectively an exploded schematic structural view and a sectional schematic structural view of a first embodiment of an immunofluorescence analysis apparatus of the present invention, as shown in fig. 1A and fig. 1B, in the first embodiment, the immunofluorescence analysis apparatus further includes a housing 10 and a first water-absorbing material 31, a first sample adding hole 11 and a mounting groove 12 which are sequentially communicated are provided on the housing 10, a black filter membrane 20 and a first water-absorbing material 31 which are stacked are provided in the mounting groove 12, wherein the black filter membrane 20 is close to the first sample adding hole 11.

Optionally, as shown in fig. 1B, the black filter membrane 20 is tightly attached to the bottom of the installation groove 12.

Optionally, the black filter membrane 20 and the first water absorbing material 31 are clamped in the mounting groove 12, the shape and size of the mounting groove 12 are adapted to the shape and size of the black filter membrane 20 and the first water absorbing material 31, for example, when the mounting groove 12 is cylindrical, the black filter membrane 20 and the first water absorbing material 31 are preferably circular.

In the first embodiment, the working principle of the immunofluorescence analysis apparatus is as follows: the sample to be analyzed is added to the black filter 20 through the first sample adding hole 11, the liquid in the sample to be analyzed and the antibody and other micro-impurities which are not combined with the object to be detected permeate through the black filter 20 and then are absorbed by the first water absorbing material 31, the object to be detected in the sample to be analyzed, which is stained by the fluorescent labeled antibody, is trapped on the surface of the black filter 20, and the trapped matter on the surface of the black filter 20 in the immunofluorescence analysis apparatus is analyzed by using a matched fluorescence detection apparatus (such as a fluorescence microscope).

Specifically, during fluorescence detection, the field of view of the fluorescence microscope must be larger than the cross-sectional area of the narrowest part of the first sample application hole 11 (e.g., the bottom of the first sample application hole 11), so that the object to be detected in the area of the black filter 20 corresponding to the narrowest part of the first sample application hole 11 can be detected by the fluorescence microscope.

The immunofluorescence analysis apparatus of the first embodiment may be named a diafiltration type immunofluorescence analysis apparatus based on the operation principle of the immunofluorescence analysis apparatus of the first embodiment.

In the first embodiment:

optionally, the first well 11 is tapered, for example conical.

Alternatively, the first well 11 is cylindrical, for example, cylindrical.

Preferably, the housing 10 is black in color to further eliminate the effect of light reflection on fluorescence detection.

Optionally, the first water absorbing material 31 is water absorbing paper, and the number of layers of the water absorbing paper is set according to needs.

Based on the immunofluorescence assay device of the first embodiment, the utility model discloses still provide an immunofluorescence assay method, include:

providing a sample to be analyzed, an immunofluorescence analysis device and a fluorescence detection device, wherein the sample to be analyzed is a mixed system comprising the sample to be analyzed and a fluorescence labeled antibody, the sample to be analyzed contains an object to be detected, and the immunofluorescence analysis device is the immunofluorescence analysis device of the first embodiment.

The sample to be analyzed is added from the first well 11 to the black filter 20 for diafiltration.

After the diafiltration, the liquid in the sample to be analyzed, the antibody not bound to the object to be detected, and other micro-impurities permeate through the black filter membrane 20, and then are absorbed by the first water-absorbing material 31, the object to be detected, which is stained by the fluorescence-labeled antibody in the sample to be analyzed, is retained on the surface of the black filter membrane 20, and the retention on the surface of the black filter membrane 20 in the immunofluorescence analysis apparatus is analyzed by using a fluorescence detection apparatus.

Fig. 2A and 2B are respectively an exploded schematic structural view and a sectional schematic structural view of a second embodiment of the immunofluorescence analysis apparatus of the present invention, as shown in fig. 2A and 2B, in the second embodiment, the immunofluorescence analysis apparatus further includes an upper cover 41 and a lower housing 42 that are connected in a mutually-matched manner, a second sample adding hole 412 is provided in the upper cover 41, the lower housing 42 has an upper cover 41 mating end and an air exhaust end that are arranged relatively, the black filter membrane 20 is fixed between the upper cover 41 and the upper cover 41 mating end of the lower housing 42, a hollow structure is provided in the lower housing 42, and the hollow structure runs through the upper cover 41 mating end and the air exhaust end.

As shown in fig. 2A and 2B, optionally, a protruding portion 421 is disposed at the mating end of the upper cover 41, the upper cover 41 is connected to the protruding portion 421 in a mating manner, and the black filter membrane 20 is fixed between the upper cover 41 and the protruding portion 421; the hollow structure comprises a first through hole 51 which is arranged at the mating end of the upper cover 41 of the lower shell 42 and penetrates through the protruding part 421, and a first groove 61 which is arranged at the air extraction end of the lower shell 42, wherein the bottom of the first groove 61 is communicated with the first through hole 51.

Alternatively, as shown in fig. 2B, the upper cover 41 is connected to the protrusion 421 in a snap-fit manner.

In the second embodiment, the working principle of the immunofluorescence analysis apparatus is as follows: connecting the immunofluorescence analysis device with an air suction device (such as a syringe), then extending one end of the immunofluorescence analysis device, which is provided with an upper cover 41, into a sample to be analyzed, enabling the air suction device to perform air suction action (such as drawing out a syringe core rod), enabling the sample to be analyzed to enter a black filter membrane 20 through a second sample adding hole 412 of the upper cover 41, enabling liquid in the sample to be analyzed, antibodies and other micro impurities which are not combined with the object to be analyzed to permeate from the black filter membrane 20 and be sucked into a hollow structure of the lower shell 42, enabling the object to be analyzed, which is stained by the fluorescence labeling antibodies, in the sample to be analyzed to be intercepted on the surface of the black filter membrane 20, and analyzing the intercepted objects on the surface of the black filter membrane 20 in the immunofluorescence analysis device by using a matched fluorescence detection device (such as a fluorescence microscope).

Specifically, in the fluorescence detection, the field of view of the fluorescence microscope must be larger than the cross-sectional area of the narrowest position (e.g., the bottom) of the second sample application hole 412, so that the object to be detected on the surface of the corresponding black filter 20 right below the second sample application hole 412 can be detected by the fluorescence microscope.

Based on the operation principle of the immunofluorescence analysis apparatus of the second embodiment, the immunofluorescence analysis apparatus of the second embodiment may be named as a suction filtration type immunofluorescence analysis apparatus.

In the second embodiment:

preferably, the color of the upper cover 41 is black, so as to further eliminate the influence of light reflection on fluorescence detection.

Specifically, the material of the lower housing 42 and the upper cover 41 may be plastic, metal, or ceramic.

Specifically, the second sampling hole 412 may have a tapered shape, such as a conical shape.

Based on the immunofluorescence assay device of the second embodiment, the utility model discloses still provide an immunofluorescence assay method, include:

providing a sample to be analyzed, an immunofluorescence analysis device, an air extraction device and a fluorescence detection device, wherein the sample to be analyzed is a mixed system comprising the sample to be analyzed and a fluorescence labeled antibody, the sample to be analyzed contains an object to be detected, and the immunofluorescence analysis device is the immunofluorescence analysis device of the second embodiment.

Connecting the immunofluorescence analysis device with an air extraction device, then extending one end of the immunofluorescence analysis device, which is provided with an upper cover 41, into a sample to be analyzed, and then enabling the air extraction device to perform air extraction action, so that the sample to be analyzed enters the black filter membrane 20 through a second sample adding hole 412 in the upper cover 41 for filtration;

after suction filtration, the liquid in the sample to be analyzed, the antibody not bound to the object to be detected and other micro impurities permeate through the black filter membrane 20 and are sucked into the cavity of the immunofluorescence analysis device, the object to be detected, which is dyed by the fluorescence labeling antibody in the sample to be analyzed, is intercepted on the surface of the black filter membrane 20, and the intercepted object on the surface of the black filter membrane 20 in the immunofluorescence analysis device is analyzed by adopting a fluorescence detection device.

It will be appreciated that in the above-described immunofluorescence assay method, since the immunofluorescence assay device is used upside down at the time of sample injection, it is preferable to reverse the immunofluorescence assay device quickly after air suction in order to prevent the sucked liquid from flowing back to the black filter 20 again, thereby ensuring the accuracy of the detection result.

Fig. 3A and 3B are schematic diagrams of an exploded structure and a schematic diagram of a cross-sectional structure of a third embodiment of an immunofluorescence analysis apparatus according to the present invention, as shown in fig. 3A and 3B, respectively, in the third embodiment, the immunofluorescence analysis apparatus further includes an upper housing 71, a fixing plug 715, a spring plate 80, and a base 72;

the upper shell 71 has a sample adding end and a base 72 mating end which are oppositely arranged, the sample adding end is provided with a second sample adding hole 412, the base 72 mating end is provided with a second groove 714, the second sample adding hole 412 is communicated with the bottom of the second groove 714, and the fixing plug 715 is fixed in the second groove 714;

the black filter membrane 20 is arranged on the end face of the fixed plug 715 close to one end of the second sample adding hole 412;

a second through hole 716 is formed in the fixing plug 715, and the second through hole 716 penetrates through one end of the fixing plug 715 close to the second sampling hole 412 and one end close to the base 72;

the base 72 comprises a bottom plate 721 and a side plate 722 connected with the bottom plate 721, an opening 723 is arranged on the bottom plate 721, the spring plate 80 is borne on the bottom plate 721, and the upper shell 71 is arranged on the spring plate 80 and connected with the base 72; a gap is formed between one end of the fixing plug 715 close to the base 72 and the elastic sheet 80.

Alternatively, as shown in fig. 3B, the black filter 20 is disposed closely to the bottom of the second groove 714.

Alternatively, as shown in fig. 3B, the upper housing 71 and the base 72 are connected by a snap connection.

The opening 723 is formed in the bottom plate 721 to facilitate an operator to press the elastic sheet 80.

In the third embodiment, the working principle of the immunofluorescence analysis apparatus is as follows:

in the immunofluorescence analysis apparatus according to the third embodiment, the upper housing 71, the fixing plug 715 and the elastic sheet 80 enclose a cavity 717, when an operator presses the elastic sheet 80 from a side of the elastic sheet 80 away from the upper housing 71, the volume of the cavity 717 decreases, air in the cavity 717 flows due to compression, and the air passes through the second through hole 716 of the fixing plug 715, the black filter 20 and the second sample adding hole 412 to be discharged outwards in sequence, so that a negative pressure is generated in the cavity 717, and a sample to be analyzed is added to the surface of the black filter 20 through the third sample adding hole 713. The depression is removed and the liquid in the sample to be analyzed can be driven from the black filter 20 into the cavity 717 by the atmospheric pressure since the cavity 717 has a negative pressure with respect to the environment. The object to be detected, which is stained with the fluorescent-labeled antibody at this time, is trapped on the surface of the black filter 20, and the liquid in the sample to be analyzed, as well as the antibody and other minute impurities that are not bound to the object to be detected, enter the cavity 717. The retentate on the surface of the black filter 20 of the immunofluorescence assay device is analyzed using a suitable fluorescence detection device (e.g., a fluorescence microscope).

Specifically, in the fluorescence detection, the field of view of the fluorescence microscope must be larger than the cross-sectional area of the narrowest position (e.g., the bottom) of the third sample application hole 713, so that the object to be detected on the surface of the corresponding black filter 20 right below the third sample application hole 713 can be detected by the fluorescence microscope.

The immunofluorescence analysis apparatus of the third embodiment may be named an air-filter type immunofluorescence analysis apparatus based on the operation principle of the immunofluorescence analysis apparatus of the third embodiment.

In the third embodiment:

preferably, the upper shell 71 is black in color to further eliminate the influence of light reflection on the fluorescence detection process.

Preferably, a second water absorbing material 32 is further fixed in the second groove 714, the second water absorbing material 32 is tightly attached to one side of the fixing plug 715 close to the base 72, a gap is provided between the second water absorbing material 32 and the elastic sheet 80, and the second water absorbing material 32 is capable of absorbing liquid flowing into the cavity 717 of the immunofluorescence analysis apparatus from the black filter membrane 20, so as to avoid backflow of the liquid to the black filter membrane 20 when the elastic sheet 80 is not pressed.

Preferably, the second water absorbent material 32 is provided with a third through hole 323 corresponding to the second through hole 716, so that the liquid flowing out of the black filter 20 can flow into the cavity 717 through the third through hole 323, and the air can flow between the cavity 717 and the second through hole 716.

Optionally, the second absorbent material 32 is absorbent paper.

Optionally, the second water absorbing material 32 is clamped in the second groove 714.

Optionally, the fixing plug 715 is fixed in the second groove 714 by a snap and adhesive method.

Optionally, the distance between one end of the fixing plug 715 close to the base 72 and the elastic sheet 80 is 2-5 mm.

Specifically, the material of the upper housing 71 and the base 72 may be plastic, metal, or ceramic.

Specifically, the third sampling hole 713 may have a tapered shape, for example, a conical shape.

Specifically, the material of the fixing plug 715 may be plastic, metal, or ceramic.

Based on the immunofluorescence assay device of the third embodiment, the utility model discloses still provide an immunofluorescence assay method, include:

providing a sample to be analyzed, an immunofluorescence analysis device and a fluorescence detection device, wherein the sample to be analyzed is a mixed system comprising the sample to be analyzed and a fluorescence labeled antibody, the sample to be analyzed contains an object to be detected, and the immunofluorescence analysis device is the immunofluorescence analysis device of the third embodiment.

The elastic sheet 80 is pressed from the side of the elastic sheet 80 away from the upper shell 71, so that a negative pressure is generated in the cavity 717, and the sample to be analyzed is added to the surface of the black filter 20 through the third sample adding hole 713 of the upper shell 71.

The pressure is removed and the ambient atmospheric pressure drives the liquid in the sample to be analyzed from the black filter 20 into the cavity 717. The object to be detected, which is stained with the fluorescent-labeled antibody at this time, is trapped on the surface of the black filter 20, and the liquid in the sample to be analyzed, as well as the antibody and other minute impurities that are not bound to the object to be detected, enter the cavity 717.

The retentate on the surface of the black filter 20 in the immunofluorescence assay device is analyzed using a suitable fluorescence detection device (e.g., a fluorescence microscope).

The immunofluorescence assay method of the present invention is described in detail below as a specific example.

Example 1

Referring to fig. 1A and 1B, embodiment 1 provides an immunofluorescence analysis method for detecting CD4 positive cells using the immunofluorescence analysis apparatus of the first embodiment (i.e., a diafiltration immunofluorescence analysis apparatus), including the following steps:

(1) mu.L of erythrocyte lysate and 1. mu.L of fluorescent dye-labeled anti-human CD4 monoclonal antibody were added to 10. mu.L of whole blood (treated with an anticoagulant), and incubated at room temperature for 15min to obtain a sample to be analyzed.

(2) And (3) adding 10 mu L of the liquid to a black filter membrane 20 in the immunofluorescence analysis device, and standing for 3-5 min.

(3) And placing the immunofluorescence analysis device in a matched fluorescence microscope, turning on a switch of the microscope, wirelessly connecting the immunofluorescence analysis device with a computer or a mobile phone, and taking a picture of the final result.

Example 2

Referring to fig. 1A and 1B, embodiment 2 provides an immunofluorescence analysis method for detecting candida albicans using the immunofluorescence analysis apparatus of the first embodiment (i.e., a diafiltration type immunofluorescence analysis apparatus), including the following steps:

(1) mu.L of PBS buffer solution and 1 mu.L of anti-Candida albicans monoclonal antibody marked by fluorescent dye are added into 10 mu.L of Candida albicans sample, and the sample to be analyzed is obtained after incubation for 15min at room temperature.

(2) And (3) adding 10 mu L of the sample to be analyzed to a black filter membrane 20 in the immunofluorescence analysis device, and standing for 3-5 min.

(3) And placing the immunofluorescence analysis device in a matched fluorescence microscope, turning on a switch of the fluorescence microscope, wirelessly connecting the fluorescence microscope with a computer or a mobile phone, and finally taking a picture of the result.

Example 3

Referring to fig. 2A and 2B, embodiment 3 provides an immunofluorescence analysis method for detecting CD4+ T lymphocytes and CD8+ T lymphocytes using the immunofluorescence analysis apparatus (i.e., the suction filtration type immunofluorescence analysis apparatus) of the second embodiment, including the following steps:

(1) mu.L of erythrocyte lysate, 0.1. mu.L of fluorochrome-labeled anti-human CD4 monoclonal antibody and 0.1. mu.L of fluorochrome-labeled anti-human CD8 monoclonal antibody are added to 10. mu.L of whole blood (treated with anticoagulant), and the mixture is incubated at room temperature for 15min to obtain a sample to be analyzed.

(2) The immunofluorescence analysis device and the syringe are connected together through a connecting piece (such as an air tube), then one end of the immunofluorescence analysis device, which is provided with the upper cover 41, extends into a sample to be analyzed, and then the syringe core rod is drawn out, so that CD4+ T lymphocytes and CD8+ T lymphocytes in the sample to be analyzed are adsorbed on the surface of the black filter membrane 20.

(3) The fluorescence pattern on the black filter 20 in the immunofluorescence assay apparatus was analyzed using a fluorescence microscope, and the ratio of CD4+ T lymphocytes and CD8+ T lymphocytes in blood was analyzed by the number of fluorescence spots.

Example 4

Referring to fig. 2A and 2B, the embodiment 4 provides an immunofluorescence analysis method for detecting candida albicans, gardnerella vaginalis and trichomonas vaginalis by using the immunofluorescence analysis apparatus (i.e., the suction filtration type immunofluorescence analysis apparatus) of the second embodiment, which includes the following steps:

(1) collecting secretion from vagina with sampling swab, putting the swab into PBS buffer, and rotating the swab in PBS buffer by hand to disperse Candida albicans, Gardnerella vaginalis and Trichomonas vaginalis in PBS buffer.

(2) Adding anti-candida albicans, gardnerella and trichomonas vaginalis antibodies which are respectively marked by green, yellow and red fluorescent dyes, uniformly mixing, standing and incubating for about 10min to obtain a sample to be analyzed.

(3) The immunofluorescence analysis device and the syringe are connected together through a connecting piece (such as an air pipe), then one end of the immunofluorescence analysis device, which is provided with an upper cover 41, extends into a sample to be analyzed, and then the syringe core rod is drawn out, so that candida albicans, gardner bacteria and trichomonas vaginalis in the sample to be analyzed are adsorbed on the surface of the black filter membrane 20.

(4) The fluorescence pattern on the black filter membrane 20 in the immunofluorescence analysis apparatus was analyzed using a fluorescence microscope, and the ratio of candida albicans, gardnerella and trichomonas vaginalis in vaginal fluid was analyzed by the number of fluorescence spots. Green fluorescent spots appearing on the black filter 20 indicate candida albicans positivity, yellow fluorescent spots indicate gardnerella positivity, and red fluorescent spots indicate trichomonas vaginalis.

Example 5

Referring to fig. 3A and 3B, embodiment 5 provides an immunofluorescence analysis method for detecting CD4+ T lymphocytes using the immunofluorescence analysis apparatus of the third embodiment (i.e., the air-filter immunofluorescence analysis apparatus), including the following steps:

(1) mu.L of erythrocyte lysate and 0.1. mu.L of fluorescent dye-labeled anti-human CD4 monoclonal antibody were added to 10. mu.L of whole blood (treated with an anticoagulant), and the mixture was incubated at room temperature for 15min to obtain a sample to be analyzed.

(2) The elastic sheet 80 in the immunofluorescence analysis device was squeezed up with a finger, and 10. mu.L of the sample to be analyzed was added to the black filter 20 in the immunofluorescence analysis device, and finally the pressure was removed.

(3) The fluorescence pattern on the surface of the black filter 20 in the immunofluorescence analyzing apparatus was analyzed by a fluorescence microscope, and the number of CD4+ T lymphocytes in blood was analyzed by the number of fluorescent spots.

Example 6

Referring to fig. 3A and 3B, embodiment 6 provides an immunofluorescence analysis method for detecting candida albicans using the immunofluorescence analysis apparatus (i.e., the air-filter immunofluorescence analysis apparatus) of the third embodiment, including the following steps:

(3) The fluorescence pattern on the surface of the black filter 20 in the immunofluorescence analysis apparatus was analyzed by a fluorescence microscope, and the number of Candida albicans was analyzed by the number of fluorescent spots.

To sum up, the utility model provides an immunofluorescence analysis device, immunofluorescence analysis device includes black filter membrane 20, immunofluorescence analysis method adopts black filter membrane 20 to hold back the object to be detected who is dyed by the fluorescence labeling antibody, filters simultaneously not with the fluorescence labeling antibody that the object to be detected combines, later adopt fluorescence detection device to the retention on black filter membrane 20 surface is analyzed, realizes reading and the count of fluorescence point; the immunofluorescence analysis method has the advantages of simple operation steps, low analysis cost and accurate detection result; the immunofluorescence analysis device is simple in structure, low in production cost, convenient to use and good in detection effect; therefore, the immunofluorescence analysis device of the utility model can be widely used, is not only suitable for laboratories of large hospitals and major, but also can be widely used in basic medical institutions.

In all examples shown and described herein, any particular value should be construed as merely exemplary, and not as a limitation, and thus other examples of example embodiments may have different values.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

The above-described embodiments are merely illustrative of several embodiments of the present invention, which are described in detail and specific, but not intended to be construed as limiting the scope of the present invention. It should be noted that, for those skilled in the art, without departing from the spirit of the present invention, several variations and modifications can be made, which are within the scope of the present invention.

Claims

1. The utility model provides an immunofluorescence analysis device, its characterized in that, includes black filter membrane, shell and first water-absorbing material, be equipped with the first application of sample hole and the mounting groove that communicate in proper order on the shell, be equipped with the black filter membrane and the first water-absorbing material of range upon range of setting in the mounting groove, wherein the black filter membrane is close to first application of sample hole sets up.

2. The immunofluorescence assay device according to claim 1, wherein the housing is black in color and the first water absorbent material is absorbent paper.

3. The utility model provides an immunofluorescence analysis device, its characterized in that, upper cover and lower casing including black filter membrane and the connection of mutually supporting, be equipped with second application of sample hole in the upper cover, the casing has the upper cover cooperation end and the end of bleeding that set up relatively down, the black filter membrane is fixed in the upper cover with between the upper cover cooperation end of casing down, the internal hollow structure that is equipped with of inferior valve, hollow structure runs through the upper cover cooperation end with the end of bleeding.

4. The immunofluorescence analysis device according to claim 3, wherein a convex portion is provided at the mating end of the upper cover, the upper cover is in mating connection with the convex portion, and the black filter membrane is fixed between the upper cover and the convex portion; the hollow structure comprises a first through hole which is arranged at the upper cover matching end of the lower shell and penetrates through the protruding part and a first groove which is arranged at the air exhaust end of the lower shell, and the bottom of the first groove is communicated with the first through hole.

5. The immunofluorescence assay device of claim 4, wherein the upper cover is snap-fit to the boss, and the upper cover is black in color.

6. An immunofluorescence analysis device is characterized by comprising a black filter membrane, an upper shell, a fixed plug, a spring plate and a base;

7. The immunofluorescence analysis device according to claim 6, wherein the upper housing is black in color, the fixing plug is fixed in the second groove in a clamping and bonding manner, and a distance between one end of the fixing plug close to the base and the elastic sheet is 2-5 mm.

8. The immunofluorescence analysis apparatus according to claim 6, wherein a second water absorbent material is further fixed in the second groove, the second water absorbent material clings to one side of the fixing plug close to the base, and a space is provided between the second water absorbent material and the elastic sheet.

9. The immunofluorescence assay device of claim 8, wherein the second bibulous material is provided with a third aperture corresponding to the second aperture.