CN208695033U - It is a kind of for extracting the micro-fluidic chip of circulating excretion body - Google Patents
It is a kind of for extracting the micro-fluidic chip of circulating excretion body Download PDFInfo
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- CN208695033U CN208695033U CN201821308561.8U CN201821308561U CN208695033U CN 208695033 U CN208695033 U CN 208695033U CN 201821308561 U CN201821308561 U CN 201821308561U CN 208695033 U CN208695033 U CN 208695033U
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Abstract
It is the utility model relates to the extraction element of excretion body, in particular to a kind of for extracting the micro-fluidic chip of circulating excretion body, belong to technical field of molecular biology.It is a kind of for extracting the micro-fluidic chip of circulating excretion body, the micro-fluidic chip includes upper layer cover plate and underlying basal, the underlying basal is equipped with reaction mixing chamber, wash buffer1 storage cavity, wash buffer2 storage cavity, sample chamber and waste liquid cavity, it is additionally provided with excretion body adsorption column on the underlying basal of micro-fluidic chip, reacts and is connected between mixing chamber and excretion body adsorption column by the first S type hole road；Wash buffer1 storage cavity and wash buffer2 storage cavity pass through the first connection duct, the second connection duct respectively and are connected with excretion body adsorption column, excretion body adsorption column is connect by duct with waste liquid cavity, and excretion body adsorption column is connected to sample chamber by the 2nd S type hole road.
Description
Technical field
It is the utility model relates to the extraction element of excretion body, in particular to a kind of for extracting the miniflow of circulating excretion body
Chip is controlled, technical field of molecular biology is belonged to.
Background technique
Excretion body (exosome) is a kind of 30-100nm size, the extracellular nanoscale cystic structures of extra-low density.Circulation
Include the important products miRNA in excretion body in blood.MiRNA (exosomal- miRNA) in serum excretion body has uniqueness
Mediated cell between the function that communicates, can specificity inhibit the expression of target gene in target cell, therefore compared with free miRNA,
The variation of serum exosomal-miRNA level can more represent the variation of the biological function of malignant tumour.Excretion body has been at present
Through being widely used in treating, diagnostic field.Therefore, it is just very crucial to obtain the stable excretion body RNA of quality.
Currently, the extracting method of excretion body be mainly summarized as it is following several: (1) filter membrane filtration method is different using aperture
The different substance of filter membrane retention relative molecular mass.Due to the cystic structures of excretion body, relative molecular mass is greater than general
Protein, therefore select different size of filter membrane that excretion body can be made to separate with other macromolecular substances.This operation is relatively
Simply, but obtained sample purity is insufficient, and may not apply to serum/plasma.(2) supercentrifugation, this is common at present
The method of extraction excretion body successively remove removing protein, cell fragment etc., finally mainly under the action of different centrifugal force
Centrifugation obtains excretion body under ultrahigh speed.The excretion scale of construction that such method obtains is more, but has purity deficiency, operating time
The shortcomings that length, the high requirements on the equipment.(3) chromatography, this method need special equipment, and application is narrow, and at high cost, application is non-
Normal is not extensive.
Utility model content
The purpose of this utility model is to provide a kind of for extracting the micro-fluidic chip of circulating excretion body, this is micro-fluidic
Chip has compact, easy to carry, only needs a small amount of sample that the advantages of extraction requires can be completed.
The technical scheme adopted by the utility model to solve the technical problem is as follows:
It is a kind of for extracting the micro-fluidic chip of circulating excretion body, which includes upper layer cover plate and lower layer's base
Piece, the underlying basal is equipped with reaction mixing chamber, wash buffer1 stores cavity, wash buffer2 stores cavity,
Sample chamber and waste liquid cavity, are additionally provided with excretion body adsorption column on the underlying basal of micro-fluidic chip, reaction mixing chamber and outer
It secretes and is connected between body adsorption column by the first S type hole road；Wash buffer1 stores cavity and wash buffer2 storage chamber
Body pass through respectively the first connection duct, second connection duct be connected with excretion body adsorption column, excretion body adsorption column pass through duct and
The connection of waste liquid cavity, excretion body adsorption column are connected to sample chamber by the 2nd S type hole road.
Preferably, the middle part of micro-fluidic chip is arranged in excretion body adsorption column, and it is located at wash buffer1 storage chamber
Between body and wash buffer2 storage cavity.
Preferably, reaction mixing chamber is located at the side of micro-fluidic chip, waste liquid cavity is located at the another of micro-fluidic chip
One side-lower, wash buffer1 storage cavity and wash buffer2 storage chamber body are located at the middle part of micro-fluidic chip.
Preferably, well is equipped on the cover plate of upper layer with the corresponding position of reaction mixing chamber, on the cover plate of upper layer with
The corresponding position of sample chamber is equipped with sample and draws hole, position corresponding with waste liquid cavity and sample chamber on the cover plate of upper layer
Equipped with negative pressure pumping holes.Negative pressure pumping holes is connected with external vacuum pump.
Preferably, position corresponding with wash buffer1 storage cavity is equipped with wash buffer1 on the cover plate of upper layer
Chamber hole is stored, position corresponding with wash buffer2 storage cavity is equipped with wash buffer2 storage chamber on the cover plate of upper layer
Body opening.
The utility model for extracting the micro-fluidic chip of circulating excretion body, each maximum can upper 800 μ l blood
Slurry samples.The excretion body adsorption column, can specificity absorption excretion body.Wash buffer1 stores cavity and wash
Cleaning solution needed for buffer2 storage cavity incorporates extraction process.Vacuum pump passes through negative pressure pumping holes and waste liquid cavity and sample
Cavity connection.
The beneficial effects of the utility model are: the function that the present apparatus incorporates separation, extracts excretion body, greatly reduces
The operation of the extraction of excretion body, the device volume is small and exquisite, easy to carry, only needs a small amount of sample that extraction can be completed and requires.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the utility model；
In figure: 1, well, 2, reaction mixing chamber, 3, wash buffer1 storage cavity, 4, wash buffer2
Storage cavity, 5, waste liquid cavity, 6, negative pressure pumping holes, the 7, the first S type hole road, 8, excretion body adsorption column, the 9, the 2nd S type hole road,
10, sample chamber, the 11, second connection duct, the 12, first connection duct, 13, sample absorption hole, 14, wash buffer1 storage
Deposit chamber hole, 15, wash buffer2 storage chamber hole.
Specific embodiment
Below by specific embodiment, and in conjunction with attached drawing, the technical solution of the utility model is made further specifically
It is bright.It should be appreciated that the implementation of the utility model is not limited by the following examples, any form that the utility model is done
On accommodation and/or change fall within scope of protection of the utility model.
In the present invention, if not refering in particular to, all parts, percentage are unit of weight, used equipment and original
Material etc. is commercially available or commonly used in the art.Method in following embodiments is unless otherwise instructed this field
Conventional method.
Embodiment:
As shown in Figure 1 is a kind of for extracting the micro-fluidic chip of circulating excretion body, which includes upper layer
Cover plate and underlying basal.
The underlying basal is equipped with reaction mixing chamber 2, wash buffer1 storage cavity 3, wash buffer2 storage
Cavity 4, sample chamber 10 and waste liquid cavity 5 are deposited, excretion body adsorption column 8 is additionally provided on the underlying basal of micro-fluidic chip.Reaction
It is connected between mixing chamber 2 and excretion body adsorption column 8 by the first S type hole road 7；Wash buffer1 storage cavity 3 passes through
First connection duct 12 is connected with excretion body adsorption column 8, wash buffer2 store cavity 4 by second connect duct 11 and
Excretion body adsorption column 8 is connected, and excretion body adsorption column 8 is connect by duct with waste liquid cavity 5, and excretion body adsorption column 8 passes through the 2nd S
Type hole road 9 is connected to sample chamber 10.
Reaction mixing chamber 2 is located at the side of micro-fluidic chip, and waste liquid cavity 5 is located under the other side of micro-fluidic chip
Side, wash buffer1 storage cavity 3 and wash buffer2 storage cavity 4 are located at the middle part of micro-fluidic chip.Excretion body is inhaled
The middle part of micro-fluidic chip is arranged in attached column 8, and is located at wash buffer1 storage cavity 3 and wash buffer2 storage cavity
Between 4.
Be equipped with well 1 on the cover plate of upper layer with the corresponding position of reaction mixing chamber 2, on the cover plate of upper layer with sample chamber
10 corresponding positions are equipped with sample and draw hole 13, and sample draws hole can the also external pipettor progress sample suction of external negative pressure pump
It takes；Position corresponding with waste liquid cavity 5 is equipped with negative pressure pumping holes 6 on the cover plate of upper layer.Vacuum pump passes through negative pressure pumping holes and waste liquid chamber
Body connection, is the retention cavity of waste liquid.Position corresponding with wash buffer1 storage cavity is equipped with wash on the cover plate of upper layer
Buffer1 stores chamber hole 14, and position corresponding with wash buffer2 storage cavity is equipped with wash on the cover plate of upper layer
Buffer2 stores chamber hole 15.
This chip is made by PC material, upper layer cover plate long 50mm, wide 20mm, thickness 2mm, the long 50mm of underlying basal,
Wide 20mm, thick 5mm.The volume of reaction mixing chamber 2 is that the volume of 1000 μ l, wash buffer1 storage cavity is 800 μ l,
The volume that wash buffer2 stores cavity is 800 μ l.
Sample Purification on Single specific steps are carried out using the micro-fluidic chip for extracting circulating excretion body of the utility model
It is specific as follows:
1. circulating Sample supernatants are added into reaction mixing chamber 2 from well 1,1h is stood；
2. opening the vacuum pump 3min connected with negative pressure pumping holes 6, the mixing of lysate and blood serum sample in mixing chamber
Object flows into waste liquid cavity 5 after the first S type hole road 7 flows into excretion body adsorption column 8；
3. opening wash buffer1 stores chamber hole 14, closed after opening the vacuum pump 1min connected with negative pressure pumping holes 6
Wash buffer1 stores chamber hole 14, and the wash buffer1 that wash buffer1 is stored in cavity 3 will be through the first connecting hole
Road 12 flows into waste liquid cavity 5 after flowing into excretion body adsorption column 8, takes away remaining ionic impurity on adsorption column；
4. opening wash buffer2 stores chamber hole 15, closed after opening the vacuum pump 1min connected with negative pressure pumping holes 6
Wash buffer2 stores chamber hole 15, and the wash buffer2 that wash buffer2 is stored in cavity 4 will be through the second connecting hole
Road 11 flows into waste liquid cavity 5 after flowing into excretion body adsorption column 8, takes away remaining big molecular impurity on adsorption column；
5. eluent is added from well 1, opens sample and draw hole 13, make its external vacuum pump, vacuumizing 30s,
Eluent enters excretion body adsorption column by the first S type hole road 7, stands 5min；
The vacuum pump 2min that hole 13 is connected is drawn with sample 6. opening, eluate sample enters sample through the 2nd S type hole road 9
Product cavity 10；
7. drawing hole 13 by sample with pipettor is sucked out eluate sample.
Embodiment described above is a kind of preferable scheme of the utility model, is not made to the utility model any
Formal limitation, there are also other variations and modifications on the premise of not exceeding the technical scheme recorded in the claims.
Claims (5)
1. a kind of for extracting the micro-fluidic chip of circulating excretion body, which includes upper layer cover plate and lower layer's base
Piece, it is characterised in that: the underlying basal is equipped with reaction mixing chamber (2), wash buffer1 stores cavity (3), wash
Buffer2 stores cavity (4), sample chamber (10) and waste liquid cavity (5), is additionally provided with excretion on the underlying basal of micro-fluidic chip
Body adsorption column (8) reacts and is connected between mixing chamber (2) and excretion body adsorption column (8) by the first S type hole road (7)；wash
Buffer1 stores cavity (3) and wash buffer2 storage cavity (4) respectively by the first connection duct (12), the second connection
Duct (11) is connected with excretion body adsorption column (8), and excretion body adsorption column (8) is connect by duct with waste liquid cavity (5), excretion body
Adsorption column (8) is connected to sample chamber (10) by the 2nd S type hole road (9).
2. according to claim 1 for extracting the micro-fluidic chip of circulating excretion body, it is characterised in that: excretion body is inhaled
The middle part of micro-fluidic chip is arranged in attached column (8), and is located at wash buffer1 storage cavity (3) and wash buffer2 storage
Between cavity (4).
3. according to claim 1 for extracting the micro-fluidic chip of circulating excretion body, it is characterised in that: reaction mixing
Cavity (2) is located at the side of micro-fluidic chip, and waste liquid cavity (5) is located at below the other side of micro-fluidic chip, wash
Buffer1 storage cavity (3) and wash buffer2 storage cavity (4) are located at the middle part of micro-fluidic chip.
4. according to claim 1 for extracting the micro-fluidic chip of circulating excretion body, it is characterised in that: upper layer cover plate
Upper position corresponding with reaction mixing chamber (2) is equipped with well (1), corresponding with sample chamber (10) on the cover plate of upper layer
Position is equipped with sample and draws hole (13), and position corresponding with waste liquid cavity (5) is equipped with negative pressure pumping holes (6) on the cover plate of upper layer.
5. according to claim 1 for extracting the micro-fluidic chip of circulating excretion body, it is characterised in that: upper layer cover plate
Upper position corresponding with wash buffer1 storage cavity is equipped with wash buffer1 and stores chamber hole (14), on the cover plate of upper layer
Position corresponding with wash buffer2 storage cavity is equipped with wash buffer2 storage chamber hole (15).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821308561.8U CN208695033U (en) | 2018-08-14 | 2018-08-14 | It is a kind of for extracting the micro-fluidic chip of circulating excretion body |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821308561.8U CN208695033U (en) | 2018-08-14 | 2018-08-14 | It is a kind of for extracting the micro-fluidic chip of circulating excretion body |
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| Publication Number | Publication Date |
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| CN208695033U true CN208695033U (en) | 2019-04-05 |
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| CN201821308561.8U Active CN208695033U (en) | 2018-08-14 | 2018-08-14 | It is a kind of for extracting the micro-fluidic chip of circulating excretion body |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111961584A (en) * | 2020-08-24 | 2020-11-20 | 山东大学齐鲁医院 | Cerebrospinal fluid exosome RNA detection device, system and method based on microfluidic technology |
-
2018
- 2018-08-14 CN CN201821308561.8U patent/CN208695033U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111961584A (en) * | 2020-08-24 | 2020-11-20 | 山东大学齐鲁医院 | Cerebrospinal fluid exosome RNA detection device, system and method based on microfluidic technology |
| CN111961584B (en) * | 2020-08-24 | 2022-05-03 | 山东大学齐鲁医院 | Cerebrospinal fluid exosome RNA detection device, system and method based on microfluidic technology |
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