CN211179859U

CN211179859U - Kit for detecting concentration of exosomes in batches

Info

Publication number: CN211179859U
Application number: CN201921858915.0U
Authority: CN
Inventors: 陈锦阳; 刘军权; 李静静
Original assignee: Zhejiang Healthfuture Biomedical Technology Co ltd
Current assignee: Zhejiang Healthfuture Biomedical Technology Co ltd
Priority date: 2019-10-31
Filing date: 2019-10-31
Publication date: 2020-08-04
Anticipated expiration: 2029-10-31

Abstract

The utility model relates to a sample detection equipment technical field specifically is a body concentration detect reagent box secretes in batches, the kit includes a thermostated container, be equipped with the layering survey board in the thermostated container, layering survey board from the top down divide into the three-layer and be reagent board, sample bottom plate and ELIAS plate bottom plate in proper order, reagent board, sample bottom plate and ELIAS plate bottom plate one corner all are equipped with the support hole, the side of reagent board, sample bottom plate and ELIAS plate bottom plate is just facing support hole department and is equipped with fixing bolt, be equipped with the survey board support in the support hole and establish ties reagent board, sample bottom plate and ELIAS plate bottom plate in proper order. The utility model discloses can carry out the whole concentration measurement of exosome fast, utilize BCA measurement principle, master the concentration parameter of the holistic exosome protein of sample.

Description

Kit for detecting concentration of exosomes in batches

Technical Field

The utility model relates to a sample test equipment technical field specifically is a exosome concentration detection kit in batches.

Background

Exosomes are in essence tiny vesicles secreted from cells, with diameters of about 30-200nm, now specifically disc-shaped vesicles with diameters of 40-100 nm; in general, all cultured cell types can secrete exosomes, and exosomes are naturally found in body fluids, including blood, saliva, urine, cerebrospinal fluid, and milk. It is mainly from the multivesicular body formed by the invagination of intracellular lysosome particles, and is released into extracellular matrix after the fusion of the outer membrane of the multivesicular body and cell membrane.

For some time, research on exosomes is an emerging focus because it carries a large number of cell signals of biological origin, cell formation, transport, etc., and its numerous functionality comes from the type of cell from which it was derived, since research on exosomes can be targeted.

Currently, the concentration of exosomes is customarily determined by instrumental detection including by Zeta potential detection, by using classical microelectrophoresis and brownian motion as modern analytical means, and the particle concentration can be analyzed by video counting. In the methods, a carrier substance such as original serum needs to be treated by using a kit, and detailed data of a specific exosome can be obtained; however, the concentration measurement and the particle size distribution of exosomes in the laboratory at present can only be performed by single operation generally, batch operation cannot be performed, the operation period is long, and the contrast data is small; on the other hand, for macroscopic exosome concentration parameters, since exosomes are complex RNAs and proteins in nature, macroscopic exosome concentration measurement does not need to adopt a complex targeted separation means, and instead, the whole concentration information is lost by this operation.

SUMMERY OF THE UTILITY MODEL

An object of the utility model is to provide an exosome concentration detection kit in batches to solve the problem that proposes in the above-mentioned background art.

In order to achieve the above object, the utility model provides a following technical scheme:

the utility model provides a volume exosome concentration detection kit, the kit includes a thermostated container, be equipped with the layering survey board in the thermostated container, the layering survey board from the top down divide into the three-layer and be reagent board, sample bottom plate and ELIAS plate bottom plate in proper order, reagent board, sample bottom plate and ELIAS plate bottom plate one corner all are equipped with the support hole, the side of reagent board, sample bottom plate and ELIAS plate bottom plate is just facing support hole department and is equipped with fixing bolt, be equipped with the survey board support in the support hole and establish ties reagent board, sample bottom plate and ELIAS plate bottom plate in proper order.

Preferably: the reagent plate is provided with a plurality of rows of unit reagent volumes, each unit cell of each row of unit reagent volumes is connected through an overflow groove, a reagent through hole penetrating through the whole reagent plate is formed in each unit reagent volume, and the number of the unit reagent volumes of each cell is 200 ul.

Preferably: and a discharge valve is traversed on the cross section of the reagent through hole, and a valve handle of the discharge valve is arranged on the end face of the left side of the reagent plate.

Preferably: the thermostat is characterized in that a timer is arranged on a box cover of the thermostat, and a temperature setting module and a prompting module are further arranged on the front side of the thermostat.

Preferably: the sample bottom plate is provided with unit sample capacity with the same interval as unit reagent capacity grids, and the capacity of each unit sample capacity is 220 ul.

Preferably: an ELISA plate limiting area is arranged on the bottom plate of the ELISA plate, and an ELISA plate is arranged in the ELISA plate limiting area.

Compared with the prior art, the beneficial effects of the utility model are that: for the overall concentration measurement of exosomes, the BCA measurement principle is utilized, the overall exosome concentration parameters of the sample are mastered, the operation is simple and rapid, and the measurement can be completed within 45 minutes; the method has the advantages of accuracy, sensitivity, good reagent stability and measurement precision of 0.5-10 ug/ml; the whole kit and the method thereof are economical to use, reagents can be injected into the upper reagent plate in batches, and the whole process can be tested in batches according to requirements, so that the macroscopic numerical value of the exosome concentration can be conveniently and rapidly mastered.

Drawings

Fig. 1 is a schematic view of the overall structure of the present invention;

FIG. 2 is a schematic structural view of a layered assay plate according to the present invention;

fig. 3 is a schematic view of the overall structure of the reagent plate of the present invention;

fig. 4 is a schematic structural diagram of a sample base plate according to the present invention;

FIG. 5 is a schematic structural view of an ELISA plate bottom plate in the present invention;

fig. 6 is a cross-sectional view of a reagent plate in the present invention;

fig. 7 is a flowchart of the measurement procedure according to the present invention.

In the figure: 1. a thermostat; 11. a timer; 12. a temperature setting module; 13. a prompt module; 2. a layered assay plate; 21. a reagent plate; 211. a bracket hole; 212. fixing the bolt; 213. unit reagent capacity; 214. an overflow trough; 215. a reagent through hole; 216. a discharge valve; 22. a sample base plate; 221. unit sample volume; 23. an ELISA plate bottom plate; 231. a limiting area of the ELISA plate; 232. an ELISA plate; 24. assay plate holder.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and to simplify the description, but do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the present invention.

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.

Referring to fig. 1-7, the present invention provides a technical solution:

the utility model provides a body concentration detect reagent box secretly in batches, the kit includes a thermostated container 1, be equipped with layering survey board 2 in the thermostated container 1, layering survey board 2 from the top down divide into the three-layer and be reagent board 21 in proper order, sample bottom plate 22 and ELIAS plate bottom plate 23, reagent board 21,

sample bottom plate

22 and 23 one corners of ELIAS plate bottom plate all are equipped with bracket hole 211, reagent board 21, the side of sample bottom plate 22 and ELIAS plate bottom plate 23 is just facing bracket hole 211 department and is equipped with fixing bolt 212, be equipped with survey board support 24 in the bracket hole 211 and with reagent board 21, sample bottom plate 22 and ELIAS plate bottom plate 23 establish ties in proper order.

The reagent plate 21 is provided with a plurality of rows of unit reagent volumes 213, each unit cell of each row of unit reagent volumes 213 is connected through an overflow groove 214, each unit reagent volume 213 is provided with a reagent through hole 215 penetrating through the whole reagent plate 21, and the volume of each unit reagent volume 213 is 200 ul.

A discharge valve 216 is provided across the cross section of the reagent passage hole 215, and a valve handle of the discharge valve 216 is provided on the left side end face of the reagent plate 21.

A timer 11 is arranged on a box cover of the constant temperature box 1, and a temperature setting module 12 and a prompting module 13 are further arranged on the front face of the constant temperature box 1.

The sample substrate 22 is provided with unit sample volumes 221 at intervals equal to 213 cells per unit reagent volume, and the number of the unit sample volumes 221 per cell is 220 ul.

The ELISA plate bottom plate 23 is provided with an ELISA plate limiting region 231, and an ELISA plate 232 is arranged in the ELISA plate limiting region 231.

The utility model discloses a work flow: referring to FIG. 7, an ELISA plate 232 is placed on the bottom plate 23 in the defined area 231 of the ELISA plate, and the ratio of BCA reagent A to BCA reagent B is 50: 1, preparing a BCA working solution; an ELISA plate bottom plate 23 and a sample bottom plate 22 are arranged and fixed on a measuring plate bracket 24, and a sample is diluted by a proper multiple to be 20ul in single sample volume; adding the BCA working solution prepared in the first step into the reagent plate 21 on the uppermost layer corresponding to the number of samples on the ELISA plate 232 and the sample base plate 22, wherein the total amount of the working solution is 200ul, discharging the working solution through the reagent through hole 215, and injecting the working solution into the ELISA plate 232 and the sample base plate 22 in batches; placing the ELISA plate 232 on a vibrator to vibrate for 30sec, and immediately placing the whole device of the ELISA plate 232 and the sample bottom plate 23 into the constant temperature box 1, setting the temperature at 37 ℃ and the time at 30 min; after 30 minutes, the enzyme label plate 232 is used as a standard reference, color comparison is carried out at the wavelength of 562nm, the light absorption value is recorded, a curve is drawn, the protein content in the exosome is inquired, and the concentration is calculated.

Furthermore, the measured concentration is divided by the total volume of the sample diluent, namely 20ul, and then multiplied by the sample dilution factor, namely the actual concentration of the exosome protein.

It should be noted that, the model and the using method of the electrical and electronic components related to the present disclosure, the timer 11 is an XH-M172 intermittent operation module, the temperature setting module 12 is a W1209 digital temperature controller of TE L ESKY brand, and the prompting module 13 is an ISD1820 voice module of TE L ESKY brand, which are all in the prior art, and the present disclosure does not relate to the improvement of the circuit and the improvement of the using method thereof, and therefore, the present disclosure can be fully understood and operated by the staff in the art, and will not be described herein again.

The foregoing shows and describes the general principles, essential features, and advantages of the invention. It should be understood by those skilled in the art that the present invention is not limited by the above embodiments, and the description in the above embodiments and the description is only preferred examples of the present invention, and is not intended to limit the present invention, and that the present invention can have various changes and modifications without departing from the spirit and scope of the present invention, and these changes and modifications all fall into the scope of the claimed invention. The scope of the invention is defined by the appended claims and equivalents thereof.

Claims

1. The utility model provides a batch exosome concentration detection kit, includes thermostated container (1), its characterized in that: be equipped with layering survey board (2) in thermostated container (1), layering survey board (2) divide into the three-layer from the top down and be reagent board (21), sample bottom plate (22) and ELIAS plate bottom plate (23) in proper order, reagent board (21), sample bottom plate (22) and ELIAS plate bottom plate (23) one corner all are equipped with support hole (211), the side of reagent board (21), sample bottom plate (22) and ELIAS plate bottom plate (23) is just facing support hole (211) department and is equipped with fixing bolt (212), be equipped with survey board support (24) in support hole (211) and establish ties reagent board (21), sample bottom plate (22) and ELIAS plate bottom plate (23) in proper order.

2. The kit for detecting the concentration of exosomes in batches according to claim 1, characterized in that: the reagent plate (21) is provided with a plurality of rows of unit reagent volumes (213), each unit cell of each row of unit reagent volumes (213) is connected through an overflow groove (214), a reagent through hole (215) penetrating through the whole reagent plate (21) is arranged in each unit reagent volume (213), and the volume number of each unit reagent volume (213) is 200 ul.

3. The kit for detecting the concentration of exosomes in batches according to claim 2, characterized in that: a discharge valve (216) penetrates through the cross section of the reagent through hole (215), and a valve handle of the discharge valve (216) is arranged on the left end face of the reagent plate (21).

4. The kit for detecting the concentration of exosomes in batches according to claim 1, characterized in that: the automatic temperature control device is characterized in that a timer (11) is arranged on a box cover of the constant temperature box (1), and a temperature setting module (12) and a prompt module (13) are further arranged on the front face of the constant temperature box (1).

5. The kit for detecting the concentration of exosomes in batches according to claim 1, characterized in that: the sample bottom plate (22) is provided with unit sample volumes (221) with the same interval as the unit reagent volume (213), and the volume number of each unit sample volume (221) is 220 ul.

6. The kit for detecting the concentration of exosomes in batches according to claim 1, characterized in that: an ELISA plate limiting region (231) is arranged on the ELISA plate bottom plate (23), and an ELISA plate (232) is arranged in the ELISA plate limiting region (231).