CN208627332U - It is a kind of for detecting the micro-fluidic chip of semaphorins 4D - Google Patents
It is a kind of for detecting the micro-fluidic chip of semaphorins 4D Download PDFInfo
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- CN208627332U CN208627332U CN201821241558.9U CN201821241558U CN208627332U CN 208627332 U CN208627332 U CN 208627332U CN 201821241558 U CN201821241558 U CN 201821241558U CN 208627332 U CN208627332 U CN 208627332U
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Abstract
A kind of for detecting the micro-fluidic chip of semaphorins 4D, including electro-conductive glass substrate and PDMS cover plate, electro-conductive glass substrate surface layer is electroplate with thin layer of indium tin oxide and forms ITO electro-conductive glass, and ITO electro-conductive glass is bonded with PDMS cover plate by oxygen gas plasma；ITO electro-conductive glass is equipped with central screening passage, and the channel surface of central screening passage is electroplate with micro- nickel column array of several micro- nickel column compositions, and micro- nickel column pan coating has semaphorins 4D antibody magnetic bead；The bottom surface of PDMS cover plate is equipped with fish bone structure corresponding with central screening passage, and fish bone structure is alternatively arranged by several grooves in " > " type with identical, and the arrow of fish bone structure is directed toward the downstream of central screening passage.Micro-fluidic chip binding signal element 4D directed screening semaphorins 4D completes the Precise Diagnosis of knot leukaemia；Micro-fluidic chip detection technique can effectively improve the detection efficiency of POCT, save detection time and diagnostic accuracy is higher.
Description
Technical field
The utility model relates to medical detection technologies, and in particular to a kind of for detecting the micro-fluidic core of semaphorins 4D
Piece.
Background technique
Microflow control technique is referred to using microchannel (having a size of tens of to hundreds of microns) processing or manipulation minute fluid
Science and Technology involved in the system of (volume be nanoliter arrive picoliters) is one and is related to chemistry, fluid physics, microelectronics, newly
The emerging cross discipline of material, biology and biomedical engineering.In actual application, it is micro-fluidic can biology, change
The basic operation units such as the, sample preparation of medical analysis process, reaction, separation, detection are integrated into one several square centimeters
On chip, it is automatically performed analysis overall process, essential characteristic and sharpest edges are a variety of monotechnicses whole controllable small
Flexible combination, scale are integrated on platform.
Leukaemia is a kind of candidate stem cell malignant clone disease.Clonal leukaemia cell because proliferation out of control, point
Change obstacle, apoptosis mechanism largely proliferation accumulation in marrow and other hematopoietic tissues such as to be obstructed, and infiltrates other non-hematopoietic tissues
And organ, while inhibiting normal hematopoiesis function.Clinical visible different degrees of anaemia, bleeding, infectious fever and liver, spleen, leaching
Fawn on enlargement and skeleton pain.It is reported that the disease incidence of China each department leukaemia accounts for the 6th in various tumours.Semaphorins
4D is the transmembrane glycoprotein of a 150kD, by 862 Amino acid profiles, extracellular conservative Sema structure receptor for identification,
Intracellular section of very short, not special functional domain, and semaphorins 4D high expression in leukemia tumor, by utilizing semaphorins
The detection of 4D can provide direct evidence for the judgement of leukaemia.
However, existing detection means is needed using numerous detection devices and cumbersome testing process, and complete white blood
Sick detection needs take a substantial amount of time and energy, it is difficult to realize detection (POCT) immediately, and detection is efficient low, is easy
The case where showing missing inspection, judging by accident.
Utility model content
The utility model to solve the above-mentioned problems, devise it is a kind of for detecting the micro-fluidic chip of semaphorins 4D, should
Micro-fluidic chip binding signal element 4D directed screening semaphorins 4D, completes the Precise Diagnosis of leukaemia；Micro-fluidic chip detects skill
Art can effectively improve the detection efficiency of POCT, save detection time and diagnostic accuracy is higher.
In order to achieve the above technical purposes, reach above-mentioned technical effect, the utility model is real by the following technical programs
Existing:
It is a kind of for detecting the micro-fluidic chip of semaphorins 4D, which is characterized in that covered including electro-conductive glass substrate and PDMS
Cover board, electro-conductive glass substrate surface layer are electroplate with thin layer of indium tin oxide and form ITO electro-conductive glass, and ITO electro-conductive glass and PDMS are covered
Plate is bonded by oxygen gas plasma；
The ITO electro-conductive glass is equipped with sample feed liquor pond, and the two sides in sample feed liquor pond are respectively equipped with the first buffer
Feed liquor pond and the second buffer feed liquor pond, ITO electro-conductive glass are equipped with central screening passage, sample feed liquor pond, the first buffer
Feed liquor pond and the second buffer feed liquor pond and central screening passage penetrate through, and the two sides of central screening passage are symmetrically arranged with several vertical
Column, the channel surface of central screening passage are electroplate with micro- nickel column array of several micro- nickel column compositions, and micro- nickel column pan coating has letter
The other end of number element 4D antibody magnetic bead, central screening passage is equipped with the sample waste liquid pool to connect with central screening passage, first
Buffer waste liquid pool and the second buffer waste liquid pool；
The PDMS cover plate upstream is equipped with perforative sample inlet, the first buffer inlet and the second buffer
Inlet, it is useless that the downstream of PDMS cover plate is equipped with perforative waste liquid liquid outlet, the first buffer waste liquid outlet and the second buffer
Liquid outlet, sample inlet, the first buffer inlet, the second buffer inlet, waste liquid liquid outlet, the first buffer waste liquid
Outlet and the sample feed liquor pond on the second buffer waste liquid outlet and ITO electro-conductive glass, the first buffer feed liquor pond, the second buffering
Liquid feed liquor pond, sample waste liquid pool, the first buffer waste liquid pool and the second buffer waste liquid pool position correspond, PDMS covering
The bottom surface of plate is equipped with fish bone structure corresponding with central screening passage, and fish bone structure is by several grooves in " > " type with identical
It is alternatively arranged, the arrow of fish bone structure is directed toward the downstream of central screening passage.
Further, the semaphorins 4D antibody magnetic bead is coated in ferroso-ferric oxide bead surface by semaphorins 4D antibody
Upper composition chondritic.
Further, the long 30mm of central screening passage, width 4mm, high 50um, the diameter of column are 500um, height
50um, the groove spacing of fish bone structure are 100~300um, and the width of groove is 50um, depth 45um.
The beneficial effects of the utility model are: the micro-fluidic chip utilizes semaphorins 4D antibody directed screening semaphorins 4D,
Complete leukaemia and the Precise Diagnosis of other tumours；Micro-fluidic chip detection technique can effectively improve the detection efficiency of POCT, section
About detection time and diagnostic accuracy is higher.
Detailed description of the invention
In order to illustrate more clearly of the technical solution of the utility model embodiment, make required for being described below to embodiment
Attached drawing is briefly described, it should be apparent that, the drawings in the following description are merely some embodiments of the present invention,
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings
Other attached drawings.
Fig. 1 is a kind of for detecting the overall structure diagram of the micro-fluidic chip of semaphorins 4D；
Fig. 2 is a kind of for detecting the Section A-A cross-sectional view of the micro-fluidic chip of semaphorins 4D；
Fig. 3 is the structural schematic diagram of the ITO electro-conductive glass；
Fig. 4 is the structural schematic diagram of the PDMS cover plate.
In attached drawing, parts list represented by the reference numerals are as follows:
1-ITO electro-conductive glass, 2-PDMS cover plate, 3- semaphorins 4D antibody magnetic bead, the center 11- screening passage, 12- are vertical
Column, the micro- nickel column of 13-, 14- sample feed liquor pond, 15- the first buffer feed liquor pond, 16- the second buffer feed liquor pond, 17- first are slow
Fliud flushing waste liquid pool, 18- sample waste liquid pool, 19- the second buffer waste liquid pool, this inlet of 21-, 22- the first buffer inlet,
23- the second buffer inlet, 24- the first buffer waste liquid outlet, 25- waste liquid liquid outlet, 26- the second buffer waste liquid go out
Mouthful, 27- passive mixer, 28- fish bone structure.
Specific embodiment
The following will be combined with the drawings in the embodiments of the present invention, carries out the technical scheme in the embodiment of the utility model
Clearly and completely describe, it is clear that the described embodiments are only a part of the embodiments of the utility model, rather than whole
Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art are without creative efforts
All other embodiment obtained, fall within the protection scope of the utility model.
It is a kind of for detecting the micro-fluidic chip of semaphorins 4D, including electro-conductive glass substrate and PDMS refering to fig. 1 shown in -4
Cover plate 2, electro-conductive glass substrate surface layer are electroplate with thin layer of indium tin oxide and form ITO electro-conductive glass 1, ITO electro-conductive glass 1 and PDMS
Cover plate 2 is bonded by oxygen gas plasma；
The ITO electro-conductive glass 1 is equipped with sample feed liquor pond 14, and it is slow that the two sides in sample feed liquor pond 14 are respectively equipped with first
Fliud flushing feed liquor pond 15 and the second buffer feed liquor pond 16, ITO electro-conductive glass are equipped with central screening passage 11, sample feed liquor pond
14, the first buffer feed liquor pond 15 and the second buffer feed liquor pond 16 are penetrated through with central screening passage 11, central screening passage 11
Two sides be symmetrically arranged with several columns 12, column 12 can prevent the PDMS cover plate of central 11 top of screening passage from collapsing；Center
The channel surface of screening passage 11 is electroplate with micro- nickel column array that several micro- nickel columns 13 form, and micro- 13 pan coating of nickel column has signal
Plain 4D antibody magnetic bead 3, the other end of central screening passage 11 be equipped with the sample waste liquid pool 18 to connect with central screening passage 11,
First buffer waste liquid pool 17 and the second buffer waste liquid pool 19；
2 upstream of PDMS cover plate is equipped with perforative sample inlet 21, the first buffer inlet 22 and second
The downstream of buffer inlet 23, PDMS cover plate is equipped with perforative waste liquid liquid outlet 25,24 and of the first buffer waste liquid outlet
Second buffer waste liquid outlet 26, sample inlet 21, the first buffer inlet 22, the second buffer inlet 23, waste liquid
Liquid outlet 25, the first buffer waste liquid outlet 24 and the second buffer waste liquid outlet 26 and the sample feed liquor on ITO electro-conductive glass
Pond 14, the first buffer feed liquor pond 15, the second buffer feed liquor pond 16, sample waste liquid pool 18,17 and of the first buffer waste liquid pool
The position of second buffer waste liquid pool 19 corresponds, and the bottom surface of PDMS cover plate 2 is equipped with fish corresponding with central screening passage
Bone structure 28, fish bone structure are alternatively arranged by several grooves in " > " type with identical, in the arrow direction of fish bone structure
The downstream of screening passage is entreated, fish bone structure can passively mix the sample liquid for flowing through central screening passage with buffer, and
Target cell namely leukaemia cell are improved in conjunction with the semaphorins 4D antibody magnetic bead in micro- nickel column, capture rate is provided.
Therein, the semaphorins 4D antibody magnetic bead 3 is coated in ferroso-ferric oxide bead surface by semaphorins 4D antibody
Upper composition chondritic.
Therein, the central screening passage 11 is 30mm long, width 4mm, high 50um, and the diameter of column 12 is 500um, height
50um, the groove spacing of fish bone structure 28 are 100~300um, and the width of groove is 50um, depth 45um.
The key of the micro-fluidic chip is the preparation of semaphorins 4D antibody magnetic bead 3, is prepared in semaphorins 4D antibody magnetic bead 3
Firstly the need of the ferroso-ferric oxide microballon being prepared into, the present embodiment prepares ferroso-ferric oxide using hydro-thermal method, and ferroso-ferric oxide
Preparation process it is as follows:
1, all experiment appliance and stirring magneton are cleaned with dilute sulfuric acid, guarantee that instrument IDE is clean, this step is quite heavy
It wants, because a small amount of impurity will cause serious pollution to last product.In addition, the liner needs of reaction kettle are individually clear
It washes.2, the Iron trichloride hexahydrate of 1.35g and 1.0g polyethylene glycol are smashed to pieces and pours into Iron trichloride hexahydrate powder in conical flask, be added
40mL ethylene glycol solution is put into stirring magneton stirring, until clear.3, by 3.6g anhydrous sodium acetate and polyethylene glycol powder
Conical flask is added in end, continues stirring and gets over 2h.4, by be stirred it is easy pour into reaction kettle, then reaction kettle is put into hydro-thermal furnace
200 DEG C of reaction 6-8h close hydro-thermal furnace after the reaction was completed and wait reaction kettle cooled to room temperature.5, by the solution in reaction kettle
Conical flask is poured into again, is then rinsed respectively 3 times with dehydrated alcohol and deionized water respectively, the when of falling cleaning solution is inhaled with magnet
The magnetic bead firmly prepared.6, the magnetic bead after having cleaned is dissolved in deionized water, ultrasonic 10min, then covers conical flask and is put into 4
It is saved in DEG C refrigerator to get ferroso-ferric oxide microballon is arrived.
It is difficult to be coated with semaphorins 4D antibody in ferroso-ferric oxide bead surface, be needed at this time to using graphene oxide to be coated with
In on ferroso-ferric oxide microballon, the coupled signal element 4D antibody on graphene oxide is as follows in coated process:
1,0.5mL ferroso-ferric oxide bead solution is taken to be added in the graphene oxide solution of 5mL, be added stirring magneton into
Row stirring 1h.2, the solution after stirring is subjected to ultrasound every 30min.3, iterative cycles repeat the operation 10 of stirring with ultrasound
It is secondary, solution to the end is obtained, is then respectively washed 3 times, is finally dissolved in deionized water, i.e., with dehydrated alcohol and deionized water
The coated ferroso-ferric oxide microballon of graphene oxide can be obtained.
After ferroso-ferric oxide bead surface has been coated with graphene oxide, Streptavidin is grafted onto bead surface, so
Biotinylated semaphorins 4D antibody is realized coupling by the combination of biotin-avidin afterwards, obtains semaphorins at this time
4D antibody magnetic bead 3, HPR fluorescent marker protein is added during coupling keeps semaphorins 4D antibody magnetic bead 3 and semaphorins 4D swollen
Oncocyte utilizes fluorescence detection device quick counter, detection after combining, semaphorins 4D antibody magnetic bead 3 is then plated on micro- nickel column
The specific detection of semaphorins 4D can be realized in 13 surface, provides foundation for the Precise Diagnosis of leukaemia.
The sample liquid of the utility model realizes microfluidic liquid pressure differential sample liquid in by NE-10001 type micro syringe pump
It is flowed in the screening passage of centre, by sample liquid with the feed liquor flow velocity of 10mL from being injected into sample inlet 21 in sample feed liquor pond 14
And along central screening passage 11, at the same using micro syringe pump with the feed liquor flow velocity of 10mL from the first buffer inlet 22 to first
Buffer is injected in buffer feed liquor pond 15, and to maintain the pH stable of sample liquid, sample liquid flows in central screening passage 11
When, the semaphorins 4D antibody of target cell, that is, leukemia tumor cells and 3 pan coating of semaphorins 4D antibody magnetic bead in sample liquid
Combine, the PDMS cover plate 2 of 11 top of central screening passage is equipped with fish bone structure, can make sample liquid passively mix make its with
Semaphorins 4D antibody sufficiently combines, and prevents missing inspection to improve the accuracy of detection；After the completion of capture, using micro syringe pump with
The feed liquor flow velocity of 15mL injects dcq buffer liquid into the second buffer feed liquor pond 16 from the second buffer inlet 23, utilizes punching
Residual signal element 4D antibody magnetic bead 3 in micro- nickel column 13 is all flushed out and enables its flow into sample waste liquid pool 18 by wash buffer
In, it is aobvious that Olympus IX81 type inversion fluorescence is provided on the connection microchannel of central screening passage 11 and sample waste liquid pool 18
Micro mirror, fluorescence microscope can semaphorins 4D antibody magnetic bead 3 to fluorescent marker and semaphorins 4D tumour cell conjugate carry out
Quickly identification and counting, to realize the care diagnostic of leukaemia, diagnosis rate is higher, more efficient.
In addition, ferroso-ferric oxide bead surface is coated with corresponding colorectal cancer antibody, breast cancer antibody, G. cephalantha
It can detect the malignant tumours such as colorectal cancer, breast cancer, G. cephalantha after the antibody proteins such as antibody.
In the description of this specification, the description of reference term " one embodiment ", " example ", " specific example " etc. means
Particular features, structures, materials, or characteristics described in conjunction with this embodiment or example are contained at least one of the utility model
In embodiment or example.The preferred embodiment in the utility model disclosed above is only intended to help to illustrate the utility model.
The detailed description of the preferred embodiment is not exhaustive, and also not limiting the utility model is only the specific reality
Apply mode.Obviously, it according to the content of this specification, can make many modifications and variations.This specification is chosen and specifically describes this
A little embodiments are in order to preferably explain the principles of the present invention and practical application, to make technical field technology people
Member can better understand and utilize the utility model.The utility model is only by claims and its full scope and equivalent
Limitation.
Claims (3)
1. a kind of for detecting the micro-fluidic chip of semaphorins 4D, which is characterized in that covered including electro-conductive glass substrate and PDMS
Plate, electro-conductive glass substrate surface layer are electroplate with thin layer of indium tin oxide and form ITO electro-conductive glass, ITO electro-conductive glass and PDMS cover plate
It is bonded by oxygen gas plasma；
The ITO electro-conductive glass is equipped with sample feed liquor pond, and the two sides in sample feed liquor pond are respectively equipped with the first buffer feed liquor
Pond and the second buffer feed liquor pond, ITO electro-conductive glass are equipped with central screening passage, sample feed liquor pond, the first buffer feed liquor
Pond and the second buffer feed liquor pond and central screening passage penetrate through, and the two sides of central screening passage are symmetrically arranged with several columns, in
The channel surface of centre screening passage is electroplate with micro- nickel column array of several micro- nickel column compositions, and micro- nickel column pan coating has semaphorins 4D
Antibody magnetic bead, the other end of central screening passage are equipped with the sample waste liquid pool, the first buffer that connect with central screening passage
Waste liquid pool and the second buffer waste liquid pool；
The PDMS cover plate upstream is equipped with perforative sample inlet, the first buffer inlet and the second buffer feed liquor
Mouthful, the downstream of PDMS cover plate is equipped with perforative waste liquid liquid outlet, the first buffer waste liquid outlet and the second buffer waste liquid and goes out
Mouthful, sample inlet, the first buffer inlet, the second buffer inlet, waste liquid liquid outlet, the first buffer waste liquid outlet
With on the second buffer waste liquid outlet and ITO electro-conductive glass sample feed liquor pond, the first buffer feed liquor pond, the second buffer into
Liquid pool, sample waste liquid pool, the first buffer waste liquid pool and the second buffer waste liquid pool position correspond, PDMS cover plate
Bottom surface is equipped with fish bone structure corresponding with central screening passage, and fish bone structure is by several grooves in " > " type with identical interval
It arranges, the arrow of fish bone structure is directed toward the downstream of central screening passage.
2. according to claim 1 a kind of for detecting the micro-fluidic chip of semaphorins 4D, which is characterized in that the letter
Number element 4D antibody magnetic bead is coated in ferroso-ferric oxide bead surface by semaphorins 4D antibody forms chondritic.
3. according to claim 1 a kind of for detecting the micro-fluidic chip of semaphorins 4D, which is characterized in that in described
The long 30mm of screening passage, width 4mm, high 50um are entreated, the diameter of column is 500um, high 50um, and the groove spacing of fish bone structure is
100~300um, the width of groove are 50um, depth 45um.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201821241558.9U CN208627332U (en) | 2018-08-02 | 2018-08-02 | It is a kind of for detecting the micro-fluidic chip of semaphorins 4D |
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| CN201821241558.9U CN208627332U (en) | 2018-08-02 | 2018-08-02 | It is a kind of for detecting the micro-fluidic chip of semaphorins 4D |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112044479A (en) * | 2019-06-05 | 2020-12-08 | 曦医生技股份有限公司 | Micro-channel device |
| CN112264115A (en) * | 2020-10-26 | 2021-01-26 | 南京鼓楼医院 | Fishbone microfluidic chip carrying molecular imprinting inverse opal structure microspheres and preparation method thereof |
| CN113295670A (en) * | 2021-05-21 | 2021-08-24 | 合肥工业大学 | Preparation method of micro-fluidic chip detection device based on SERS substrate |
-
2018
- 2018-08-02 CN CN201821241558.9U patent/CN208627332U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112044479A (en) * | 2019-06-05 | 2020-12-08 | 曦医生技股份有限公司 | Micro-channel device |
| CN112264115A (en) * | 2020-10-26 | 2021-01-26 | 南京鼓楼医院 | Fishbone microfluidic chip carrying molecular imprinting inverse opal structure microspheres and preparation method thereof |
| CN113295670A (en) * | 2021-05-21 | 2021-08-24 | 合肥工业大学 | Preparation method of micro-fluidic chip detection device based on SERS substrate |
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