CN216187231U - Immune cell function assessment kit for healthy people - Google Patents
Immune cell function assessment kit for healthy people Download PDFInfo
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- CN216187231U CN216187231U CN202122471770.2U CN202122471770U CN216187231U CN 216187231 U CN216187231 U CN 216187231U CN 202122471770 U CN202122471770 U CN 202122471770U CN 216187231 U CN216187231 U CN 216187231U
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Abstract
The utility model discloses a kit for evaluating immune cell functions of healthy people, which comprises a box body (1) and a box cover (2) which is connected with the box body (1) and can be opened and closed, wherein the box body is enclosed by a front side plate (3), a right side plate (4), a rear side plate (5), a left side plate (6) and a bottom plate (7); be provided with buffering collet (8) in box body (1), be equipped with in proper order along left side board (6) to right side board (4) direction on buffering collet (8) and be used for the antibody to place the district and supplementary district of placing, the antibody is placed and is equipped with the antibody container in the district and places hole (9), supplementary placing is equipped with the solvent container in the district and places hole (10). The utility model has simple structure and reasonable size, is made of light materials and is convenient to carry; the arrangement of the antibodies is sequentially and annularly arranged according to the antibody channels and the number of the tubes, the design is reasonable, the antibodies can be taken conveniently, and the working efficiency is improved.
Description
Technical Field
The utility model belongs to the technical field of kits, and particularly relates to a kit for evaluating immune cell functions of healthy people.
Background
The kit is a box for containing chemical reagents for detecting chemical components, drug residues, virus species and the like. Is generally used by hospitals and pharmaceutical enterprises. The kit is provided with a complete set of all reagents necessary for analysis or determination, the operation is convenient, and the error of artificially configuring the medicines is eliminated, because all the reagents are present in the complete set, the operation is more standard, and the waste phenomenon is avoided.
Only when the immune function of a person is kept in a normal state, viruses, germs, dead cells, mutant cells and the like can be identified, eliminated and eliminated, so that diseases are prevented from being induced. Immune dysfunction (referred to as hyperactivity, decline or deficiency) can give rise to pathogenic factors, and induce various diseases, such as common cold, pneumonia, tuberculosis, rheumatoid diseases, inflammation of liver, gallbladder and kidney, gastrointestinal diseases, and even cancer. Therefore, maintaining normal immune function is the fundamental guarantee for preventing diseases and keeping health. The function detection and evaluation of the autoimmune system are carried out at proper time, which is helpful for evaluating the immune function state of the organism, thereby keeping healthy immunity against various diseases.
However, through the research literature at home and abroad, it is found that no method for comprehensively evaluating the immune function of healthy people exists internationally at present. At present, the methods for evaluating the human body immune function by clinical and third-party detection institutions at home and abroad still stay in the conventional indexes such as the number, the proportion and the like of cells. For example, hospital blood routinely gives only a range and ratio of the number of leukocytes, such as the number, ratio of CD3 cells, the number, ratio of CD4+ T cells, and the like. These data are of little reference to the clinical diagnosis of the physician. At present, no method for truly evaluating immune function exists in clinical and third-party detection institutions.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide an immune cell function assessment kit for comprehensively assessing healthy people.
The technical scheme adopted by the utility model is as follows:
a kit for evaluating immune cell functions of healthy people comprises a box body (1) and a box cover (2) which is connected with the box body (1) and can be opened and closed, wherein the box body is enclosed by a front side plate (3), a right side plate (4), a rear side plate (5), a left side plate (6) and a bottom plate (7); be provided with buffering collet (8) in box body (1), be equipped with in proper order along left side board (6) to right side board (4) direction on buffering collet (8) and be used for the antibody to place the district and supplementary district of placing, the antibody is placed and is equipped with the antibody container in the district and places hole (9), supplementary placing is equipped with the solvent container in the district and places hole (10).
As a further improvement of the technical scheme, a first row of placing holes, a second row of placing holes, a third row of placing holes, a fourth row of placing holes and a fifth row of placing holes are sequentially arranged in the antibody placing area along the direction from the left side plate (6) to the right side plate (4), the first row of placing holes and the second row of placing holes are respectively provided with 7 antibody container placing holes (9), the third row of placing holes is provided with 8 antibody container placing holes (9), the fourth row of placing holes is provided with 7 antibody container placing holes (9), and the fifth row of placing holes is provided with 4 antibody container placing holes (9).
As a further improvement of the technical scheme, the auxiliary placing area is provided with 2 solvent container placing holes (10) along the direction from the front side plate (3) to the rear side plate (5).
As a further improvement of the technical scheme, the antibody container placing hole (9) and the solvent container placing hole (10) are both provided with soft elastic depressions for clamping the reagent bottles on the buffer bottom supports (8).
As a further improvement of the technical scheme, the diameter of the antibody container placing hole (9) is 0.8-1.2 cm.
As a further improvement of the technical scheme, the diameter of the solvent container placing hole (10) is 3-4 cm.
As a further improvement of the technical scheme, the antibody container placing hole (9) is used for containing antibodies marked by different fluorescent markers.
As a further improvement of the above technical solution, the solvent container placing hole (10) is used for containing different concentrated reagents.
As a further improvement of the technical scheme, the box body (1) and the box cover (2) are made of hard paperboards.
As a further improvement of the technical scheme, the material of the buffering bottom support (8) is pearl cotton.
As a further improvement of the technical scheme, the immune cells of the healthy population are T cells, B cells or NK cells.
The utility model has the beneficial effects that:
the immune cell (T cell, B cell and NK cell) function evaluation kit for healthy people adopts multiple immune function indexes to comprehensively evaluate the immune function in peripheral blood of the healthy people, the indexes can comprehensively reflect the immune function states (cellular immunity, humoral immunity and cytotoxicity reaction) of the healthy people to prevent major diseases, the quantitative parameters of an immune system are utilized to effectively evaluate the immunity of sub-healthy people, the unbalanced individuals of the immune system are intervened in advance, disease development signs such as autoimmune diseases and cancers can be discovered earlier than clinic, the early prediction and prevention capability of the diseases is improved, early discovery, early prevention and early intervention are really achieved, the occurrence and development of the major diseases are prevented, and the goal of preventing the diseases is achieved. And can combine flow cytometer to be used for carrying out comprehensive assessment to healthy crowd's immunologic function, and the testing process easy operation, low cost can carry out large-scale detection simultaneously, uses manpower sparingly and time, improves work efficiency.
The reagent bottle is simple in structure and convenient to carry, is designed aiming at the reagent bottle required by function evaluation, and is provided with the placing hole and the buffering bottom support, and the reagent bottle is reasonable in size, made of light materials and convenient to carry; the arrangement of the antibodies is sequentially and annularly arranged according to the antibody channels and the number of the tubes, the design is reasonable, the antibodies can be taken conveniently, and the working efficiency is improved.
Drawings
Fig. 1 is a schematic view of the structure of the packing box of the present invention.
Figure 2 is a schematic view of the package opening configuration of the present invention.
Fig. 3 is a schematic view of the structure of the buffering shoe of the present invention.
In the figure: (1) a box body; (2) and a box cover; (3) a front side plate; (4) a right side plate; (5) a rear side plate; (6) a right side plate; (7) a bottom plate; (8) the buffer bottom support; (9) antibody container placing holes; (10) and a solvent container placing hole.
Detailed Description
The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.
Example 1
As shown in fig. 1-3, a kit for evaluating immune cell (T cell, B cell, NK cell) function of healthy people comprises a box body (1), and an openable box cover (2) connected to the box body (1), wherein the box body is enclosed by a front side plate (3), a right side plate (4), a rear side plate (5), a left side plate (6) and a bottom plate (7); be provided with buffering collet (8) in box body (1), be equipped with in proper order along left side board (6) to right side board (4) direction on buffering collet (8) and be used for the antibody to place the district and supplementary district of placing, the antibody is placed and is equipped with the antibody container in the district and places hole (9), and the antibody container is used for holding by the antibody of different fluorescence marker marks, and supplementary district of placing is equipped with a plurality of solvent containers and places hole (10), and the solvent container is used for holding different concentrated reagent.
The antibody container placing hole (9) and the solvent container placing hole (10) are both provided with a buffering bottom support (8) and soft elastic depressions for clamping the reagent bottles.
The antibody placing area is internally provided with a first row of placing holes, a second row of placing holes, a third row of placing holes, a fourth row of placing holes and a fifth row of placing holes in sequence along the direction from a left side plate (6) to a right side plate (4), wherein the first row of placing holes and the second row of placing holes are respectively provided with 7 antibody container placing holes (9), the third row of placing holes is provided with 8 antibody container placing holes (9), the fourth row of placing holes is provided with 7 antibody container placing holes (9), the fifth row of placing holes is provided with 4 antibody container placing holes (9), and the antibody container placing holes (9) are all grouped according to color matching combination panel according to the following experiment operation and the working principle of a flow cytometer. The antibody containers are all brown plastic tubes that are completely opaque, that is, have zero light transmittance.
The arrangement of the antibodies is sequentially and annularly arranged according to the antibody channels and the number of the tubes, and each group is arranged from left to right in the following table, see table 1, so that the antibodies are convenient for operators to take and are not easy to disorder.
TABLE 1 antibody assignment sequence
The auxiliary placing area is provided with 2 solvent container placing holes (10) along the direction from the front side plate (3) to the rear side plate (5), and solvents are erythrocyte lysate (10X) reagent bottles and PBS buffer solution (10X) reagent bottles respectively.
The diameter of the antibody container placing hole (9) is 1cm, and the diameter of the solvent container placing hole (10) is 3.5 cm.
The box body (1) and the box cover (2) are made of hard paperboards, and the buffering bottom support (8) is made of pearl cotton.
A kit for evaluating the functions of immune cells (T cells, B cells and NK cells) of a healthy population can be combined with a flow cytometer to detect and analyze the T cells, the B cells and the NK cells of the healthy population to obtain 39 immune function indexes, wherein the 39 immune function indexes comprise:
total T cell percentage, total T cell count, helper T cell percentage, helper T cell count, inhibitory T cell percentage, inhibitory T cell count, helper T cell/inhibitory T cell ratio, regulatory T cell percentage, γ δ T cell percentage, V δ 1 cell percentage, V δ 2 cell percentage, V δ 1 cell/V δ 2 cell ratio, CD4+ initial T cell percentage, CD4+ initial T cell count, CD4+ effector and memory T cell percentage, CD4+ effector and memory T cell count, CD8+ initial T cell percentage, CD8+ initial T cell count, CD8+ effector and memory T cell percentage, CD8+ effector and memory T cell count, depleted CD4+ T cell percentage, depleted CD8+ T cell percentage, activated total T cell percentage, follicular helper T cell percentage, inactivated T cell percentage, and T cell percentage, Percentage of Th1 cells, percentage of Th2 cells, ratio Th1/Th2, percentage of helper T cells 17, percentage of total B lymphocytes, total B lymphocyte counts, percentage of memory B cells, percentage of plasma cells, percentage of naive B cells, percentage of NK cells, count of NK cells, percentage of NK-T cells, percentage of activated NK cells, percentage of immature NK cells, percentage of mature NK cells.
In order to obtain the above 39 immune function indexes, firstly, Peripheral Blood Mononuclear Cells (PBMCs) are extracted from human peripheral blood, then a certain amount of antibodies in the kit are respectively taken, all antibodies are mixed and incubated with the peripheral blood mononuclear cells to obtain a cell detection sample, then a flow cytometer is used for detecting the cell detection sample, and analysis software carried by the flow cytometer or third-party analysis software is used for carrying out statistical analysis on analysis results of various immune cells to obtain the above 39 immune function indexes.
Specifically, the cell markers of each immune cell population are:
total population of T cells: CD3 +;
helper T cell: CD3+ CD4 +;
inhibitory T cells: CD3+ CD8 +;
regulatory T cells: CD4+ CD25+ CD127 +;
γ δ T cells: CD3+ HLA-DR-TCR γ δ +;
initializing CD4+ T cells: CD3+ CD4+ CD45RA +;
effector memory CD4+ T cells: CD3+ CD4+ CD45RO +;
initializing CD8+ T cells: CD3+ CD8+ CD45RA +;
effector memory CD8+ T cells: CD3+ CD8+ CD45RO +;
depleted CD4+ T cells: CD3+ CD4+ CD 28-;
depleted CD8+ T cells: CD3+ CD8+ CD 28-;
terminal senescent CD8+ T cells: CD3+ CD8+ CD28-CD57 +;
activation of total T cells: CD3+ HLADR +;
follicular helper Tfh cells: CD3+ CD4+ CXCR5+ PD-1 +;
th1 cells: CD3+ CD4+ CCR6-CCR4-CXCR3+ CCR 10-;
th2 cells: CD3+ CD4+ CCR6-CCR4+ CXCR 3-;
th17 cells: CD3+ CD4+ CCR4+ CCR6+ CCR10-CXCR 3-;
total B cell population: CD19 +;
memory B cells: CD19+ CD27 +;
plasma cell: CD19+ IgD-CD20+ CD 27-;
naive B cells: CD19+ IgD + CD20+ CD 27-;
NK cells: CD3-CD56 +;
NKT cells: CD3+ CD56 +;
activating NK cells: CD3-CD56+ NKG2D +;
immature NK cells: CD3-CD56+ bright;
mature NK cells: CD3-CD56+ dim.
The utility model also provides a method for evaluating the functions of immune cells (T cells, B cells and NK cells) of healthy people, which comprises the following steps:
1) taking a 15mL centrifuge tube, and marking a corresponding sample on the centrifuge tube;
2) putting 700 μ L of whole blood sample into a centrifuge tube, adding 6300 μ L of erythrocyte lysate into the centrifuge tube, mixing well, performing lysis for 5-10min, and turning upside down every 3min during the mixing;
3) after the solution is clear and bright, centrifuging for 5min by a horizontal centrifuge at a centrifugal force of 400 Xg;
4) discarding the supernatant, taking no care to pour out the excessive precipitate, adding 7mL of PBS solution, and sucking and beating uniformly;
5) centrifuging at 400 Xg centrifugal force for 5min with horizontal centrifuge;
6) the supernatant was discarded, and the excess pellet was discarded, followed by resuspension with 700. mu.L of PBS solution.
1) taking out corresponding antibodies from a refrigerator at 4 ℃ according to an antibody list required by all color schemes of the kit, and placing the corresponding antibodies on a prepared ice bag for later use;
2) taking a 15mL centrifuge tube, taking the dosage of the color scheme list as a single test dosage, and premixing the antibody of each panel in advance according to the dosage (N +1 or N +2) of the actual test (actual test number N) N;
3) adding all antibodies in each panel into the same centrifugal tube according to the actual test dosage, sucking and beating the antibodies by using a gun head, and uniformly mixing the antibodies or performing vortex oscillation on the antibodies (after the vortex oscillation, performing instantaneous centrifugation on the antibodies);
4) sieving the cells obtained in the step 1.6 by using a cell screen;
5) subpackaging 100 mu L of the sieved cells in 15mL centrifuge tubes;
6) adding all the antibodies in the prepared panel according to the dosage of the antibodies in the cell color matching panel, and incubating for 15min in a dark place
7) Resuspend and mix well with 1mL PBS solution
8) The cells were centrifuged at 400 Xg for 5min in a horizontal centrifuge, the supernatant was discarded, resuspended in 300. mu.L of PBS solution and transferred to a flow tube.
And 3, detecting and analyzing the cell detection sample by adopting a flow cytometer:
1) opening the instrument and checking the condition of the instrument
2) Establishing a template: starting a computer, starting a Software BD FACS Diva Software to wait for machine connection, and if the Software does not have a template required by a user, reestablishing one Software, wherein the specific steps are as follows:
a. establishing a New Folder-New extreme t-New specification-New Tube (sample name) in sequence under an administeror under a Browser window;
b. if the detected content is the same as before, the former template can be copied and then the date is changed;
c. selecting a scatter diagram tool in a tool bar, wherein the abscissa of the scatter diagram tool is FSC-A, and the ordinate of the scatter diagram tool is SSC-A;
d. clicking the selected graphic type in the toolbar, pulling a mouse to form a picture, and changing the name of a coordinate axis according to a receiving channel required by the fluorescent dye;
e. set gate according to your experimental control sample, select the different shapes of gate in the toolbar, will gather in the figure and get the conglomerate cell to select. If a Marker line needs to be set, clicking a transverse line or a square ellipse in a toolbar and the like, and marking the transverse line in the histogram; selecting a cross or other square ellipses and irregular figures from the scatter diagram as a mark;
f. after the template is established, the test sample can be assayed.
3) Exporting data
a. Selecting an experiment needing to be exported by a right key, and selecting Export > FCS File;
selection FCS2.0 below flowjo10.0; FCS2.0 or FCS3.0 or FCS3.1> > OK selectable for FlowJo10.0 or more;
c. selecting a saving path;
d. PDF export saving is also possible.
The present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other without conflict.
Claims (8)
1. A kit for evaluating immune cell functions of healthy people comprises a box body (1) and a box cover (2) which is connected with the box body (1) and can be opened and closed, wherein the box body is enclosed by a front side plate (3), a right side plate (4), a rear side plate (5), a left side plate (6) and a bottom plate (7); be provided with buffering collet (8) in box body (1), be equipped with in proper order along left side board (6) to right side board (4) direction on buffering collet (8) and be used for the antibody to place the district and supplementary district of placing, the antibody is placed and is equipped with the antibody container in the district and places hole (9), supplementary placing is equipped with the solvent container in the district and places hole (10).
2. The kit for evaluating immune cell function of healthy population according to claim 1, wherein a first row of placing holes, a second row of placing holes, a third row of placing holes, a fourth row of placing holes and a fifth row of placing holes are sequentially formed in the antibody placing region along the direction from the left side plate (6) to the right side plate (4), wherein 7 antibody container placing holes (9) are formed in each of the first row of placing holes and the second row of placing holes, 8 antibody container placing holes (9) are formed in the third row of placing holes, 7 antibody container placing holes (9) are formed in the fourth row of placing holes, and 4 antibody container placing holes (9) are formed in the fifth row of placing holes.
3. The kit for evaluating immune cell function of healthy population according to claim 1, wherein said auxiliary placement area is provided with 2 solvent container placement holes (10) along the direction from front plate (3) to rear plate (5).
4. The kit for evaluating immune cell function of healthy people according to claim 1, wherein the antibody container placing hole (9) and the solvent container placing hole (10) are both provided with a soft elastic recess for clamping the reagent bottle on the buffer base (8).
5. The kit for evaluating immune cell function of healthy people according to any one of claims 1 to 4, wherein the antibody container placement hole (9) has a diameter of 0.8 to 1.2 cm.
6. The kit for evaluating immune cell function of healthy population according to any one of claims 1 to 4, wherein the solvent container placement hole (10) has a diameter of 3 to 4 cm.
7. The kit for evaluating immune cell function of healthy people according to any one of claims 1 to 4, wherein the material of the box body (1) and the box cover (2) is hard cardboard.
8. The kit for evaluating immune cell function of healthy people according to any one of claims 1 to 4, wherein the material of the buffering base (8) is pearl cotton.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202122471770.2U CN216187231U (en) | 2021-10-13 | 2021-10-13 | Immune cell function assessment kit for healthy people |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202122471770.2U CN216187231U (en) | 2021-10-13 | 2021-10-13 | Immune cell function assessment kit for healthy people |
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| CN216187231U true CN216187231U (en) | 2022-04-05 |
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- 2021-10-13 CN CN202122471770.2U patent/CN216187231U/en active Active
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