CN207130273U - A kind of magnetic force micro-fluidic chip for capturing bacterium - Google Patents
A kind of magnetic force micro-fluidic chip for capturing bacterium Download PDFInfo
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- CN207130273U CN207130273U CN201720925886.XU CN201720925886U CN207130273U CN 207130273 U CN207130273 U CN 207130273U CN 201720925886 U CN201720925886 U CN 201720925886U CN 207130273 U CN207130273 U CN 207130273U
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Abstract
The utility model discloses a kind of magnetic force micro-fluidic chip for capturing bacterium, it includes detection chip, sheet glass and magnet fixed chip, the detection chip is arranged on the top of the sheet glass, and the detection chip and the sheet glass are irreversible sealing, the magnet fixed chip is arranged on the lower section of the sheet glass, and the magnet fixed chip and the sheet glass are reversible sealing；The detection chip is arranged with least one sense channel in parallel；The magnet fixed chip includes magnet-fixation grooves and magnet, the magnet-fixation grooves are used to fix the magnet, magnet fixed chip is connected using reversible sealing-in mode, the magnet fixed chip repetitive cycling below sheet glass is set to use, simplify becomes the design and making of complicated micro-fluidic chip because magnetic field is intervened, micro-fluidic chip design is simple, makes, be easy to process, while can realize quickly high flux detection sensitive to cell, bacterium, virus etc. using the micro-fluidic chip.
Description
Technical field
The utility model belongs to biological technical field, and in particular to a kind of magnetic force micro-fluidic chip for capturing bacterium.
Background technology
Microfluidic chip technology is bases such as biology, chemistry, the sample preparation of medical analysis process, reaction, separation, detections
This operating unit is integrated on the chip of one piece of micro-meter scale, is automatically performed analysis total system, because it is in biology, chemistry, doctor
The great potential in etc. field, have been developed as a biology, chemistry, medical science, fluid, electronics, material, mechanical subject and intersect
Brand-new research field.Microfluidic analysis chip is by microchannel, Micropump, micro-valve, micro- reservoir, micro- by Micrometer-Nanometer Processing Technology
The function element such as electrode, micro- detecting element, window and connector makes them be integrated on chip material as integrated circuit
Micro-total analysis system, also referred to as chip lab.
In the prior art, in order to improve the specificity of traditional microfluidic chip and sensitivity, on micro-fluidic chip directly
Hole, plunger or array of fixed microparticle etc., complicated, the cost height of existing micro-fluidic chip are set.
Utility model content
For overcome the deficiencies in the prior art, it is micro- that the purpose of this utility model aims to provide a kind of magnetic force for capturing bacterium
Fluidic chip.
What the technical solution of the utility model was realized in：
The utility model embodiment provides a kind of magnetic force micro-fluidic chip for capturing bacterium, and the device includes detection
Chip, sheet glass and magnet fixed chip, the detection chip are arranged on the top of the sheet glass, and the detection chip
It is irreversible sealing with the sheet glass, the magnet fixed chip is arranged on the lower section of the sheet glass, and the magnet
Fixed chip and the sheet glass are reversible sealing；The detection chip is arranged with least one sense channel in parallel；The magnetic
Ferropexy chip includes magnet-fixation grooves and magnet, and the magnet-fixation grooves are used to fix the magnet.
In such scheme, the sense channel includes injection port, sample intake passage, sensing chamber, the sample output passage being sequentially communicated
And outlet.
Preferably, the sample intake passage is linear pattern, and the size of the sample intake passage is 10000 μm of 200 μ m；Institute
It is linear pattern to state sample output passage, and the size of the sample output passage is 15000 μm of 200 μ m；The sensing chamber is racetrack.
Preferably, the magnet-fixation grooves are correspondingly arranged with the sensing chamber.
Preferably, the material of the detection chip and the magnet fixed chip is dimethyl silicone polymer, poly- methyl
One kind in methyl acrylate, makrolon, polystyrene, polypropylene, polyethylene.
Compared with prior art, the utility model embodiment provides a kind of magnetic force micro-fluidic chip for capturing bacterium, its
Including detection chip, sheet glass and magnet fixed chip, the detection chip is arranged on the top of the sheet glass, and described
Detection chip and the sheet glass are irreversible sealing, and the magnet fixed chip is arranged on the lower section of the sheet glass, and
The magnet fixed chip and the sheet glass are reversible sealing；It is logical that the detection chip is arranged with least one detection in parallel
Road；The magnet fixed chip includes magnet-fixation grooves and magnet, and the magnet-fixation grooves are used to fix the magnet, so,
Magnet fixed chip is connected using reversible sealing-in mode, the magnet fixed chip repetitive cycling below sheet glass is used,
Simplify becomes the design and making of complicated micro-fluidic chip because magnetic field is intervened, and makes the making micro-fluidic chip without carrying out
Passage and room pre-treatment, micro-fluidic chip design is simple, makes, be easy to process, while can be realized using the micro-fluidic chip
The high flux of cell, bacterium, virus etc. is detected.
Brief description of the drawings
Fig. 1 is a kind of structural representation for the magnetic force micro-fluidic chip for capturing bacterium of the utility model embodiment；
Fig. 2 is that a kind of CAD of the detection chip for the magnetic force micro-fluidic chip for capturing bacterium of the utility model embodiment is set
Meter figure；
Fig. 3 is a kind of magnet fixed chip for the magnetic force micro-fluidic chip for capturing bacterium of the utility model embodiment
CAD design figure.
Fig. 4 is a kind of catching for the embodiment 1 of the preparation method for magnetic force micro-fluidic chip that the utility model embodiment provides
Obtain rate result figure.
Reference is as follows：
1 --- --- --- sample intake passage, 13 --- sensing chamber, 14 --- go out sample and led to for injection port, 12 for detection chip, 11
--- --- --- magnet fixed chip, 31 --- the magnet-fixation grooves, 32 --- magnet of sheet glass, 3 of outlet, 2 of road, 15.
Embodiment
In order that the purpose of this utility model, technical scheme and advantage are more clearly understood, below in conjunction with accompanying drawing and implementation
Example, the utility model is further elaborated.It should be appreciated that specific embodiment described herein is only explaining
The utility model, it is not used to limit the utility model.
The utility model embodiment provides a kind of magnetic force micro-fluidic chip for capturing bacterium, as shown in figure 1, it includes inspection
Survey chip 1, sheet glass 2 and magnet fixed chip 3, the detection chip 1 and be arranged on the top of the sheet glass 2, and it is described
Detection chip 1 and the sheet glass 2 are irreversible sealing, and the magnet fixed chip 3 is arranged on the lower section of the sheet glass 2,
And the magnet fixed chip 3 and the sheet glass 2 are reversible sealing；The detection chip 1 is arranged with parallel at least one
Sense channel；The magnet fixed chip 3 includes magnet-fixation grooves 31 and magnet 32, and the magnet-fixation grooves 31 are used to fix institute
State magnet 32.
Wherein, the sense channel includes injection port 11, sample intake passage 12, sensing chamber 13, the sample output passage being sequentially communicated
14 and outlet 15.
Wherein, the sample intake passage 12 is linear pattern, and the size of the sample intake passage 12 is 10000 μm of 200 μ m；
The sample output passage 14 is linear pattern, and the size of the sample output passage 14 is 15000 μm of 200 μ m；The sensing chamber 13
For racetrack.
The magnet-fixation grooves 31 are correspondingly arranged with the sensing chamber 13.
The material of the detection chip 1 and the magnet fixed chip 3 is dimethyl silicone polymer, polymethylacrylic acid
One kind in methyl esters, makrolon, polystyrene, polypropylene, polyethylene.
Compared compared with micro-fluidic chip, the utility model mainly has following technical advantage：Connected using reversible sealing-in mode
Connect magnet fixed chip, the magnet fixed chip repetitive cycling below sheet glass is used, simplify because magnetic field intervention and
Become the design and making of complicated micro-fluidic chip, the making micro-fluidic chip is entered row of channels and room pre-treatment, should
Micro-fluidic chip design is simple, makes, be easy to process, while can be realized to cell, bacterium, virus etc. using the micro-fluidic chip
Sensitive quickly high flux detection.
It is as follows that the utility model embodiment provides a kind of preparation process for the magnetic force micro-fluidic chip for capturing bacterium：
Step 1, detection chip lithography mask version is prepared：Detection chip mask plate is drawn according to detection chip CAD design figure,
Detection chip CAD design figure in silicon chip surface using spin-coating method as shown in Fig. 2 uniformly coat the heating that photoresist is placed in 95 DEG C
10min front baking processing is toasted on flat board, is exposed on the silicon chip detection chip mask plate being placed in after the front baking
Handle, silicon chip is placed on 95 DEG C of heating flat board to the rear baking processing for toasting 5min after exposure, the silicon chip after the rear baking is entered
Row development treatment, it is placed in developer solution after 3min, the photoresist for removing silicon chip surface is rinsed with absolute ethyl alcohol again, rinses use well
High pure nitrogen dries up, and obtains detection chip lithography mask version；
Step 2, magnet fixed chip lithography mask version is prepared：Magnet is drawn according to magnet fixed chip CAD design figure to consolidate
Determine chip mask plate, magnet fixed chip CAD design figure in silicon chip surface using spin-coating method as shown in figure 3, uniformly coat photoetching
Glue is placed in toasting 10min front baking processing on 95 DEG C of heating flat board, the magnet fixed chip mask plate is placed in described
Exposure-processed is carried out on silicon chip after front baking, silicon chip is placed in on 95 DEG C of heating flat board the rear baking for toasting 5min after exposure-processed
Processing, development treatment is carried out to the silicon chip after the rear baking, is placed in developer solution after 3min, removes the photoresist of silicon chip surface again
Rinsed with absolute ethyl alcohol, rinse well and dried up with high pure nitrogen, obtain magnet fixed chip lithography mask version；
Step 3, PDMS mixed liquors are configured：PDMS A liquid and PDMS B liquid are pressed 10:1 ratio is well mixed, and is obtained
PDMS mixed liquors；
Step 4, detection chip is prepared：The inspection that the PDMS mixed liquors casting step 1 obtained using the step 3 is obtained
Chip lithography mask version is surveyed, is removed using vavuum pump after air inside the PDMS mixed liquors and carries out drying and processing, dried
80 DEG C, drying time 30min of temperature, the cutting acquisition detection chip after the solidification of PDMS mixed liquors；
Step 5, magnet fixed chip is prepared：The PDMS mixed liquors casting step 2 obtained using the step 3 is obtained
Magnet fixed chip lithography mask version, remove after air inside the PDMS mixed liquors and carry out at drying using vavuum pump
Reason, 80 DEG C, drying time 30min of drying temperature, the cutting acquisition magnet fixed chip after the solidification of PDMS mixed liquors；
Step 6, sealing-in magnetic force micro-fluidic chip：It is clear that detection chip and sheet glass are placed in plasma in plasma cleaner
Wash 30 seconds, plasma processing is carried out to the upper surface of sheet glass, detection chip and sheet glass upper surface are subjected to irreversible envelope
Connect, the magnet fixed chip that the step 5 is obtained carries out reversible sealing-in with sheet glass lower surface, obtains the micro-fluidic core of magnetic force
Piece.
Embodiment 1
Capture pneumonia streptococcus is carried out using a kind of magnetic force micro-fluidic chip for being used to capture bacterium provided by the utility model
Bacterium, its detailed process are as follows：
Step 1：Seal magnet fixed chip：By the irreversible top for being packaged on sheet glass of detection chip, magnet is fixed
The reversible lower section for being packaged on sheet glass of chip, make sensing chamber's phase of the magnet and the detection chip in the magnet fixed chip
It is corresponding；Afterwards, 1% bovine serum albumin(BSA) is used to be carried out with 10 μ L/min flow velocity to the passage of the detection chip at closing
Reason；
Step 2：Surface antibody modification, the immune magnetic after being modified are carried out to immunomagnetic beads using pneumococci antibody
Pearl, the immunomagnetic beads after modification is passed through to the passage of the detection chip with 10 μ L/min flow velocity, makes the immune magnetic
Magnet is fixed described in integument；Afterwards, passage of the phosphate buffer solution with 10 μ L/min flow velocity to the detection chip is used
First time cleaning is carried out, to remove loose magnetic bead in passage；
Step 3：Fluorescent staining processing, the institute after control fluorescent staining processing are carried out to streptococcus pneumonia bacteria suspension to be measured
State streptococcus pneumonia bacteria suspension to be measured and the immunomagnetic beads after modification is flowed through with 5 μ L/min flow velocity；Afterwards, using phosphoric acid
Salt buffer solution carries out second to the passage of the detection chip with 10 μ L/min flow velocity and cleaned, and is not exempted from removing by described
The bacterium of epidemic disease magnetic capture；Remove the magnet fixed chip below sheet glass, detected using inverted fluorescence microscope described in
The fluorescence signal of immunomagnetic beads, the fluorescence signal that streptococcus pneumonia to be measured occurs in the immunomagnetic beads is pneumonia streptococcus to be measured
Bacterium is captured by immunomagnetic beads.
EHEC is captured using above-mentioned micro-fluidic chip, tested as a comparison, is specially：To be measured big
The uncommon bacterium bacteria suspension of intestines angstrom carries out fluorescent staining processing, the EHEC bacteria suspension to be measured after control fluorescent staining processing
The immunomagnetic beads of the surface modification pneumococci antibody after modification is flowed through with 5 μ L/min flow velocity；Afterwards, using phosphoric acid
Salt buffer solution carries out second to the passage of the detection chip with 10 μ L/min flow velocity and cleaned, and is not exempted from removing by described
The bacterium of epidemic disease magnetic capture；Remove the magnet fixed chip below sheet glass, detected using inverted fluorescence microscope described in
The fluorescence signal of immunomagnetic beads, the fluorescence signal that EHEC to be measured occurs in the immunomagnetic beads are that large intestine angstrom to be measured is uncommon
Bacterium is captured by immunomagnetic beads.
In addition, the effect that bacterium is captured to the micro-fluidic chip carries out quantitative calculating, by streptococcus pneumonia bacteria suspension and greatly
The uncommon bacterium bacteria suspension of intestines angstrom is passed through the micro-fluidic chip, collects the bacteria suspension before being captured through magnetic force micro-fluidic chip and through magnetic force miniflow
The bacteria suspension after chip capture is controlled, and Bacteria Culture is carried out using plating method respectively, by calculating through the micro-fluidic core of magnetic force
The ratio of the bacterium colony number of the front and rear bacteria suspension of piece capture, draws capture rate of the magnetic force micro-fluidic chip to streptococcus pneumonia,
Capture rate calculation formula is as follows：
A kind of capture rate result figure of the embodiment 1 of the preparation method of magnetic force micro-fluidic chip provided by the utility model,
As shown in figure 4, using micro-fluidic chip provided by the utility model, the immunomagnetic beads of surface modification streptococcus pneumonia is passed through, is divided
It is other that capture test is carried out to streptococcus pneumonia and EHEC, as seen from the figure, when bacteria suspension concentration to be measured is respectively
1.5×106CFU/mL、1.5×105CFU/mL、1.5×104CFU/mL、1.5×103CFU/mL、1.5×102CFU/mL、1.5
×101During CFU/mL, the micro-fluidic chip reaches more than 90% to the capture rate of streptococcus pneumonia, and to EHEC
Capture rate infinite approach 0%, when illustrating that the immunomagnetic beads for being passed through surface modification streptococcus pneumonia carries out bacterium capture, this is micro-fluidic
Chip only captures streptococcus pneumonia, and EHEC is not almost captured, it is seen then that carries out bacterium using the micro-fluidic chip
Capture has efficiently specificity and sensitivity, meanwhile, improve antigen-antibody joint efficiency.
In summary, captured using a kind of magnetic force micro-fluidic chip for being used to capture bacterium provided by the utility model
Bacterium and the comparing result for using traditional microfluidic chip capture bacterium, it is as shown in the table：
| Magnetic force micro-fluidic chip detects | Traditional detection method | |
| Detection time | Within 2 hours | 2 days or more than 2 days |
| Bacteria Culture condition | Without culture | Condition of culture is harsh |
| Take antibiotic influences on it | Without influence | As a result may false negative |
| Cost | It is low | It is high |
| Detect place | It is portable | It need to complete to detect in laboratory |
It can be seen that compared with the method for carrying out capturing bacterium using traditional microfluidic chip, the utility model mainly has following
Technical advantage：Using the magnet fixed chip of reversible sealing-in, make detection process operation simpler；Magnetic bead table is completed outside chip
Chip is passed through again after the antibody modification of face, avoids antibody from being adsorbed in the non-detection region in chip, is advantageous to solid phase carrier again and is immunized
The batch making of magnetic bead in vitro；Immunomagnetic beads after modification can preserve a period of time in 4 degree of refrigerators, be advantageous to using
Chip sensing chamber, the magnetic bead that can take preservation at any time are passed through in chip, further reduce the time of detection sample；Avidin
Biotinylated antibody is fixed on magnetic bead surfaces by magnetic bead, antibody is firmly combined with magnetic bead, moreover it is possible to ensures the antigen binding of antibody
Site is preferably combined with antigen, improves antigen-antibody joint efficiency；The side of bacterium capture is carried out using the micro-fluidic chip
Method improves the efficient specificity of detection, sensitivity, the consumption for reducing sample and reagent, the pollution for reducing external environment condition, and
And the repetition experiment that energy multichannel is automatical and efficient, meanwhile, based on the hydrodynamics under minute yardstick, the detection method association reaction
Accelerate, detection time shortens, and may also reach good capture rate for the sample of rareness.
It is described above, preferred embodiment only of the present utility model, it is not intended to limit protection of the present utility model
Scope.
Claims (5)
1. a kind of magnetic force micro-fluidic chip for capturing bacterium, it is characterised in that fixed including detection chip, sheet glass and magnet
Chip, the detection chip is arranged on the top of the sheet glass, and the detection chip is irreversible with the sheet glass
Sealing, the magnet fixed chip are arranged on the lower section of the sheet glass, and the magnet fixed chip and the sheet glass
For reversible sealing；The detection chip is arranged with least one sense channel in parallel；The magnet fixed chip is consolidated including magnet
Determine groove and magnet, the magnet-fixation grooves are used to fix the magnet.
2. a kind of magnetic force micro-fluidic chip for capturing bacterium according to claim 1, it is characterised in that the detection is logical
Road includes injection port, sample intake passage, sensing chamber, sample output passage and the outlet being sequentially communicated.
3. a kind of magnetic force micro-fluidic chip for capturing bacterium according to claim 2, it is characterised in that the sample introduction leads to
Road is linear pattern, and the size of the sample intake passage is 10000 μm of 200 μ m；The sample output passage is linear pattern, and institute
The size for stating sample output passage is 150000 μm of 200 μ m；The sensing chamber is racetrack.
4. a kind of magnetic force micro-fluidic chip for capturing bacterium according to claim 2, it is characterised in that the magnet is consolidated
Determine groove to be correspondingly arranged with the sensing chamber.
A kind of 5. magnetic force micro-fluidic chip for capturing bacterium according to claim 1, it is characterised in that the detection core
The material of piece and the magnet fixed chip is dimethyl silicone polymer, polymethyl methacrylate, makrolon, polyphenyl second
One kind in alkene, polypropylene, polyethylene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720925886.XU CN207130273U (en) | 2017-07-28 | 2017-07-28 | A kind of magnetic force micro-fluidic chip for capturing bacterium |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720925886.XU CN207130273U (en) | 2017-07-28 | 2017-07-28 | A kind of magnetic force micro-fluidic chip for capturing bacterium |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109306323A (en) * | 2017-07-28 | 2019-02-05 | 香港大学深圳医院 | A kind of magnetic force micro-fluidic chip and preparation method thereof for capturing bacterium |
| CN109735430A (en) * | 2019-01-28 | 2019-05-10 | 武汉纺织大学 | A kind of micro-fluidic chip of three-dimensional magnetophoresis separation |
| CN110596107A (en) * | 2019-09-23 | 2019-12-20 | 中国人民解放军陆军军医大学第一附属医院 | Reusable high-flux terahertz metamaterial rapid detection method |
-
2017
- 2017-07-28 CN CN201720925886.XU patent/CN207130273U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109306323A (en) * | 2017-07-28 | 2019-02-05 | 香港大学深圳医院 | A kind of magnetic force micro-fluidic chip and preparation method thereof for capturing bacterium |
| CN109735430A (en) * | 2019-01-28 | 2019-05-10 | 武汉纺织大学 | A kind of micro-fluidic chip of three-dimensional magnetophoresis separation |
| CN110596107A (en) * | 2019-09-23 | 2019-12-20 | 中国人民解放军陆军军医大学第一附属医院 | Reusable high-flux terahertz metamaterial rapid detection method |
| CN110596107B (en) * | 2019-09-23 | 2020-12-11 | 中国人民解放军陆军军医大学第一附属医院 | Reusable high-flux terahertz metamaterial rapid detection method |
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