CN209656719U - A kind of chemiluminescence immunoassay system - Google Patents
A kind of chemiluminescence immunoassay system Download PDFInfo
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- CN209656719U CN209656719U CN201920286137.6U CN201920286137U CN209656719U CN 209656719 U CN209656719 U CN 209656719U CN 201920286137 U CN201920286137 U CN 201920286137U CN 209656719 U CN209656719 U CN 209656719U
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Abstract
The utility model discloses a kind of chemiluminescence immunoassay systems, including liquid-transfering device, temperature control device, magnetic separation device, optical detection device, sample reagent storage device and the electricity to moisten micro-fluidic chemiluminescence detection chip.Electricity moistens micro-fluidic chemiluminescence detection chip according to electric infiltration phenomenon, apply the electrode voltage sequence being pre-designed by the electrod-array at the die substrate back side, so that the drop of chip surface is moved between each functional areas of chip surface according to scheduled path, and drop separation can be carried out in functional areas and mixes operation.Liquid-transfering device is used for from sample reagent storage device mobile reagent to filling area, and from waste liquid memory block removes waste liquid.Compared to traditional equipment, the cost of the batch detection of multisample or the joint-detection of the more reagents of single sample can be really realized on one chip.
Description
Technical field
The utility model relates to chemiluminescence detection fields, and in particular to a kind of chemiluminescence immunoassay system.
Background technique
Chemiluminescence is the light radiation generated by chemical reaction.Chemiluminescence analysis has high sensitivity, selection
Property it is preferable, instrument is simple, analysis speed is fast, the range of linearity can wide the advantages that reaching several orders of magnitude, in environment, life, medicine etc.
Field obtains more and more extensive application.The essential distinction of chemiluminescence and other luminesceence analyses is that system generates (the light spoke that shines
Penetrate) the energy source that is absorbed is different.System generates chemiluminescence, it is necessary to have one generates the light radiation reaction that can examine signal
The chemical reaction for leading to the independent reaction step of the enough energy of luminescence phenomenon can be once provided with one.
Currently, the micromation of chemiluminescence diagnostic products, is typically based on micro-fluidic chip.Microfluidic chip technology
It (Microfluidics) is basic operations such as biology, chemistry, the sample preparation of medical analysis process, reaction, separation, detections
Unit is integrated on the chip of one piece of micro-meter scale, is automatically performed analysis overall process.Micro-fluidic chip uses similar semiconductor
Micro electro mechanical processing technology constructs microflow path system on chip, by experiment with analytic process reprint to by communication with one another path and
On the chip structure of liquid phase cell composition, after loading biological sample and reaction solution, using micromechanics pump, electric hydraulic pump and electroosmotic flow
The methods of the flowing of buffer in driving chip, microfluidic circuit is formed, in carrying out a kind of or continuous a variety of reaction on chip.Laser
A variety of detection systems such as induced fluorescence, electrochemistry and chemistry and many detection means in conjunction with the analysis means such as mass spectrum are
It is used in micro-fluidic chip, quick, accurate and high throughput analysis is carried out to sample.The maximum feature of micro-fluidic chip is one
The micro-total analysis system of the numerous compound system of Multifunctional centralized architectonical sum number mesh can be formed on a chip.Microreactor is
In chip lab commonly be used for biochemical reaction structure, as Capillary Electrophoresis, polymerase chain reaction, enzyme reaction and
The microreactor etc. of DNA hybridization reaction.Therefore, chip often designs complexity, higher cost, is difficult to mass production.
Magnetic bead refers to that magnetic nano-particle formed in conjunction with inorganic or organic molecule is dispersed in certain base fluid
Colloidal state composite material with high stability.Since magnetic bead has a magnetic responsiveness, at low cost, less energy consumption and pollution-free etc. special
Point, people in magnetic bead surfaces or by the functional group of magnetic bead surfaces, such as amino, carboxyl, sulfydryl and ethylene oxide, by enzyme,
The bioactive substances such as antibody, oligonucleotides are fixed, and can be further used for enzyme immobilizatio, target medicine carrier, cell
Sorting, immune detection, albumen and nucleic acid isolate and purify and the fields such as hybridization check.Traditional immunology detection is mostly with ELISA Plate
For solid phase carrier, suspension magnetic bead as carrier specific surface area with higher, can substantially more with example reaction, in addition
The flexible Application of externally-applied magnetic field has higher sensitivity, faster detection speed compared with ELISA Plate carrier and preferably repeats
Property the advantages that, be widely used at present biology and medicine detection etc. fields.
Summary of the invention
Goal of the invention: being directed to the above-mentioned prior art, propose a kind of chemiluminescence immunoassay system, reduces a kind of chemistry hair
The design and preparation cost of light immunoassay system.
Technical solution: a kind of chemiluminescence immunoassay system, including liquid-transfering device, temperature control device, magnetic separation device,
Optical detection device, sample reagent storage device and the electricity moisten micro-fluidic chemiluminescence detection chip；Wherein:
It includes the cavity formed by bottom plate and cover board that the electricity, which moistens micro-fluidic chemiluminescence detection chip, the cavity quilt
Several functional areas are divided into, the functional areas include filling area, cleaning Disengagement zone, incubation reaction area, light detection area, waste liquid storage
Area；Connection path is equipped between the functional areas, the plate upper surface where the functional areas and the connection path is laid
Electrod-array, the electrod-array surface coat hydrophobic/super-hydrophobic coat；
The liquid-transfering device is used for from the sample reagent storage device mobile reagent to the filling area, and from institute
It states waste liquid memory block and removes waste liquid；
The temperature control device is used to control the temperature in the incubation reaction area；
Separation control is carried out to magnetic bead the magnetic separation device is for carrying out magnetic bead cleaning in the cleaning Disengagement zone when；
The optical detection device is used to collect the photon issued when light detection chemiluminescence reaction.
Further, the cover board of filling area's face is equipped with filler port, before the top of the filler port is equipped with filling
Processing component, the filling pre-treatment component include the filling channel for being connected to the filling area, are equipped with and inhale in the filling channel
It is attached to use filler assembly.
Further, the unit plane of each functional areas is adjusted by the way that the local height of each functional areas of cover board face is arranged
The corresponding liquid containing amount of product.
Further, the position in light detection area described in the cover board face is set as convex lens structures.
Further, the quantity of the filler port is greater than 1.
Further, the absorption with filler assembly be polyester film, glass fibre element film, cellulosic filter paper, in non-woven fabrics
One or more materials be made.
Further, the absorption is using microballon made of hydrophilic material, and on the microballon with filler assembly
Connection is for combining the conjugate of molecule to be filtered.
Further, the filling channel vertically connects the cover board, is equipped with atmospheric equilibrium in the filling channel side
Through-hole, the atmospheric equilibrium through-hole are located at below the position of the absorption filler assembly.
The utility model has the advantages that the chemiluminescence immunoassay system of the utility model moistens micro-fluidic chemiluminescence detection using electricity
Chip, chip infiltrate (electowetting) phenomenon according to electricity, apply the electricity being pre-designed by the electrod-array at the bottom plate back side
Pole tension sequence, so that the drop of chip surface is moved between each functional areas of chip surface according to scheduled path,
And drop separation can be carried out in functional areas and mix operation, compared to traditional micro-fluidic chip, chip surface does not have to set
Pipeline, the Micropump/micro-valve of meter and preparation complexity, structure is simple, designs and prepares cost and is far below traditional micro-fluidic chip.
Pass through relative to traditional chemiluminescence immunoassay system since chip surface eliminates complicated pipeline, Micropump/micro-valve
The batch detection of multisample or the joint-detection of the more reagents of single sample can be really realized in conjunction with liquid-transfering device, and based on tradition
The chemiluminescence immunoassay system of micro-fluidic chip this function only theoretically may be implemented, in actual design and preparation
When the micro-fluidic chip of such function, often design difficulty is big, and reagent encapsulation requires high, technique difficulty to realize, to lack certain
Practicability.
Detailed description of the invention
Fig. 1 is chemiluminescent analyzer structural schematic diagram；
Fig. 2 is the chip surface structural schematic diagram for the more reagent joint-detections of single sample；
Fig. 3 is the chip surface structural schematic diagram for multisample batch detection；
Fig. 4 is filling one schematic diagram of pre-treatment component working method；
Fig. 5 is filling two schematic diagram of pre-treatment component working method；
Fig. 6 is the structural schematic diagram of cover board face light detection zone position；
Fig. 7 is this chemiluminescent analyzer single testing process schematic diagram.
Specific embodiment
Further explanation is done to the utility model with reference to the accompanying drawing.
As shown in Figure 1, a kind of chemiluminescent analyzer, including the inspection of liquid-transfering device 5, temperature control device, magnetic separation device, light
It surveys device 4, sample reagent storage device 6 and electricity and moistens micro-fluidic chemiluminescence detection chip 7.Wherein:
It includes the cavity formed by bottom plate and cover board that electricity, which moistens micro-fluidic chemiluminescence detection chip 7, and cavity is divided into
Several functional areas, functional areas include filling area, cleaning Disengagement zone, incubation reaction area, light detection area, waste liquid memory block, by setting
The corresponding liquid containing amount of the unit area for setting the local height of each functional areas of cover board face to adjust each functional areas.Functional areas it
Between be equipped with connection path, plate upper surface where functional areas and connection path lays hydrophobic/super-hydrophobic coat, hydrophobic/super
Electrod-array is laid under hydrophobic coating.Usually select copper as electrode material, electrode size is generally 1.5mm × 1.5mm, core
Piece baseboard material is typically chosen FR4.
The cover board for filling area's face is equipped with several filler ports, and the top of filler port is equipped with filling pre-treatment component, such as
Shown in Fig. 2, Fig. 3, filler port has ten.As shown in Figure 4, Figure 5, filling pre-treatment component includes that the filling in connection filling area is logical
Road 1 fills the vertical connecting cover plate in channel 1, fills and is equipped with absorption filler assembly 2 in channel, is equipped in filling channel side big
Gas balances through-hole 3, and atmospheric equilibrium through-hole 3 is located at below the position of absorption filler assembly 2.
Filling of the filler port for sample, reagent, excitation reagent, cleaning solution etc..Absorption is used for absorption sample with filler assembly
Specific components in this, such as: the interception filtering for the above particulate matter of the micron orders such as haemocyte or fibrin aggressiveness can be with
It is made, is also possible to above-mentioned using one of polyester film, glass fibre element film, cellulosic filter paper and non-woven fabrics or multiple material
Coagulant, haemocyte membrane antibody are added in material；For the interception of certain specific molecular, the hydrophilies material such as agarose can be used
Microballon made of expecting, and conjugate of the connection for combination molecule to be filtered on microballon.The surrounding of filler assembly 2 can be embedded in
1 side wall of channel is filled, or is supported by there is the porous structure of some strength.As shown in Figure 4, Figure 5, sample adds from top
It adds in filling channel 1, under conditions of gravity or external application air pressure, sample can penetrate into filler assembly 2, therein
Specific components can be adsorbed by filler assembly 2, and the group branch for being more suitable for detection in addition flows into after passing through filler assembly 2 from filling
Hole enters in the filling area on bottom plate, realizes the pre-treatment and filling of sample.
As shown in Figure 2 and Figure 3, connection path is equipped between each functional areas, connection path can be the side of several unit areas
The continuous path of block composition, is correspondingly arranged an electrode in the corresponding lower surface of base plate of the square of each unit area.Each function
Area can be equally made of the square of several unit areas, and same its is correspondingly arranged on electrod-array.Similarly, each functional areas
And the electrode of multiple array arrangements can also be correspondingly arranged under the square of the unit area of continuous path, so as to improve drop shifting
The precision or efficiency of dynamic, mixing etc..
Droplet size is determined by the height of electrode area and cavity in chip, one timing of electrode area, single electrode area
Corresponding droplet size increases with housing depth and is become larger, therefore can be accommodated more by tuning up housing depth when chip design
More drops, to reduce chip area, the height of general cavity is no more than 0.5mm.
Liquid-transfering device is used for the mobile reagent from sample reagent storage device and removes to filling area, and from waste liquid memory block
Waste liquid.Liquid-transfering device hardware is mainly made of sample needle, fluid line, pump valve and movement mechanism, and movement mechanism drives sample needle
Carry out horizontal and elevating movement, filling area and waste liquid memory block in operation area Covering samples reagent storage means, chip.Fortune
Motivation structure drives sample needle to be moved to sample reagent storage device and draws a certain amount of sample or reagent, generally in 5ul or more, so
It is moved to above the filler port of chip afterwards, the liquid of absorption is discharged, realize the first of sample, reagent, excitation reagent, cleaning solution etc.
Step distribution, and waste liquid in the chip of waste liquid memory block is emptied with liquid-transfering device.
Temperature control device is located at the incubation reaction area back side of chip, and incubation reaction area temperature is controlled the temperature value in setting.
Temperature control device can be using the heating device being mounted on instrument, such as resistance wire or semiconductor heat electric refrigerator etc. and temperature
The closed-loop control system that sensor is formed is to guarantee temperature stable on chip；Heating device and temperature sensor can also be integrated
It, can more accurate effective control temperature although chip cost increased in this way on chip.Chemiluminescence incubation reaction
Temperature general control at 37 ± 0.5 DEG C.
Magnetic separation device carries out carrying out separation control to magnetic bead when magnetic bead cleaning for cleaning in Disengagement zone.Separating mechanism
Positioned at the cleaning Disengagement zone back side of chip.Its hardware body be permanent magnet, by adjusting the distance between chip of permanent magnet,
Or the power on/off by electromagnet, the magnetic field of control chip cleaning Disengagement zone are strong and weak.For the adsorption effect for guaranteeing magnetic bead, usually select
Ndfeb magnet or electromagnet are taken, magnetic field strength is generally 4000 Gauss.
As shown in fig. 6, the position in the cover board face light detection area of chip is set as convex lens structures, convenient for aggregation light letter
Number.Optical detection device 4 is located above the light detection area of chip, for collecting the light issued when the reaction of light detection chemiluminescence
Son.Light detection generally uses the photoelectric devices such as photomultiplier tube, photocell as light signal collection element.
Sample reagent storage device is mainly used for storing sample and reagent, when necessary containing temperature control system with guarantee sample and
The storage temperature of reagent requires, or guarantees the mixing of heterogeneous reagent containing mechanical reciprocation device.This sample reagent
Storage device can place a certain number of samples and kit, to meet continuous detection, the entry detection of instrument system.
Single testing process includes sample-adding, liquid relief, mixing, incubation, six cleaning separation, light detection steps, such as Fig. 7 institute
Show, specifically:
1) it is loaded: required sample, reagent, silicone oil etc. is filled into the filling area of chip by liquid-transfering device.When sample-adding
Further include online Sample pretreatment step, i.e., after sample is equipped with filling pre-treatment component by the top of filler port, component occurs
Separation so that be more suitable for detection sample components stay in filling area bottom electrode above.
2) liquid relief: the phenomenon that being infiltrated according to electricity, hydrophobic/super-hydrophobic coat of chip surface is passed through high pressure in corresponding electrode
After electricity, hydrophobic/super-hydrophobic coat can become than more hydrophilic, and the capillary phenomenon generated using surface tension, liquid can flow to connection
The electrode of high pressure.By a series of electrode voltage sequence of pre-programmeds, liquid can be mobile in chip surface by scheduled path.It is logical
The contact potential series of design pressor areas electrode is crossed, drop needed for experiment being isolated from the sample or reagent in filling area, or
Drop in each functional areas is moved to required position.
3) mix: isolated drop can be in chip top with procedure script, i.e., predefined electrode voltage sequence definition
Mode move.Procedure script only needs sample/reagent needed for reaction being moved to same position, they will merge automatically
At a bigger drop, then even separates and merge again again in different directions reciprocating motion, pitch of the laps operation, so that it may is real
Now mix.
4) incubate: control drop is moved to incubation reaction area and is sufficiently reacted, after the reaction time for reaching setting, by liquid
Drop is moved to cleaning Disengagement zone.
5) cleaning separation: when separation, control magnet, which moves upwards, to be close to chip lower layer or controls electromagnet be powered, by magnetic bead
It is adsorbed on chip surface, control drop removes cleaning Disengagement zone, and under extremely strong magnetic field, small magnetic bead will be gathered into very
Close magnet, and cannot be taken away by drop, thus realize that magnetic bead is separated with unreacted dissociant.When cleaning, control
Magnet moves downward or controls electromagnet power-off, and magnetic field disappears, and mobile cleaning drop is mixed with the magnetic bead after separating, by magnetic
The unreacted dissociant being mingled in pearl is discharged into cleaning drop.Repeated isolation, cleaning process will test in drop not
The dissociant of reaction sufficiently washs.
6) light detection: mobile excitation reagent, excitation reagent is usually exciting liquid or substrate, after exciting reagent droplet and cleaning
Magnetic bead combine and mix, the drop after mixing is moved to light detection area, the photon that acquisition reaction issues.
7) drop that detection is completed is moved to waste liquid memory block.
Electricity moistens micro-fluidic chemiluminescence detection chip and needs according to the more reagent joint-detections of single sample or multisample batch
Amount detection carrys out the connection path between the position of each functional areas and size in design chips, and each functional areas of design.Entry
When joint-detection or multisample batch detection:
1) sample S and reagent R1, R2, R3, R4, R5 are filled from sample reagent storage device to the filling area of chip, such as
Shown in Fig. 1；Or sample S1, S2, S3, S4, S5, S6, S7, S8 and reagent R fill adding to chip from sample reagent storage device
Area is infused, as shown in Figure 2.Reagent R1 is generally made of 3 kinds of reagent components, and component 1 is R1-1, and component 2 and 3 can be added to simultaneously
One filler port forms R1-23.
2) according to time beat, such as 20s, come carry out one by one liquid relief, mix complete detection drop TEST1, TEST2,
TEST3。
3) drop after mobile mixing incubates scheduled duration to incubation reaction area；
4) after reaching incubative time, mobile drop to cleaning Disengagement zone, using formed after the cleaning of W drop TEST1 ',
TEST2',TEST3'；W drop is cleaning reagent.
5) separation T drop forms TEST1 ", TEST2 ", TEST3 " with the detection droplets mixing after cleaning, and moves respectively
Light detection is carried out to detection zone；T drop is to excite reagent.
6) drop that detection is completed is moved to waste liquid memory block.
Above-mentioned steps 2), 4), 5) in the number of drops that is formed according to detection type, sample number it is different and different.
The above is only the preferred embodiment of the utility model, it is noted that for the common skill of the art
For art personnel, without departing from the principle of this utility model, several improvements and modifications can also be made, these improve and
Retouching also should be regarded as the protection scope of the utility model.
Claims (10)
1. a kind of chemiluminescence immunoassay system, which is characterized in that including liquid-transfering device, temperature control device, magnetic separation device,
Optical detection device, sample reagent storage device and electricity moisten micro-fluidic chemiluminescence detection chip；Wherein:
It includes the cavity formed by bottom plate and cover board that the electricity, which moistens micro-fluidic chemiluminescence detection chip, and the cavity is divided
At several functional areas, the functional areas include filling area, cleaning Disengagement zone, incubation reaction area, light detection area, waste liquid memory block；
Connection path is equipped between the functional areas, the plate upper surface where the functional areas and the connection path lays electrode
Array, the electrod-array surface coat hydrophobic/super-hydrophobic coat；
The liquid-transfering device is used for the mobile reagent from the sample reagent storage device and gives up to the filling area, and from described
Remove waste liquid in liquid memory block；
The temperature control device is used to control the temperature in the incubation reaction area；
Separation control is carried out to magnetic bead the magnetic separation device is for carrying out magnetic bead cleaning in the cleaning Disengagement zone when；
The optical detection device is used to collect the photon issued when light detection chemiluminescence reaction.
2. chemiluminescence immunoassay system according to claim 1, it is characterised in that: the cover board of filling area's face
It is equipped with filler port, the top of the filler port is equipped with filling pre-treatment component, and the filling pre-treatment component includes connection institute
The filling channel in filling area is stated, is equipped with absorption filler assembly in the filling channel.
3. chemiluminescence immunoassay system according to claim 1, it is characterised in that: by the way that the cover board face is arranged
The local height of each functional areas is come the corresponding liquid containing amount of unit area that adjusts each functional areas.
4. chemiluminescence immunoassay system according to claim 1, it is characterised in that: the inspection of light described in the cover board face
The position for surveying area is set as convex lens structures.
5. chemiluminescence immunoassay system according to claim 2, it is characterised in that: the quantity of the filler port is greater than
1。
6. chemiluminescence immunoassay system according to claim 2 or 5, it is characterised in that: absorption filler group
Part is made of one of polyester film, glass fibre element film, cellulosic filter paper, non-woven fabrics or multiple material.
7. chemiluminescence immunoassay system according to claim 2 or 5, it is characterised in that: absorption filler group
Part is that connection is used to combine the conjugate of molecule to be filtered using microballon made of hydrophilic material, and on the microballon.
8. chemiluminescence immunoassay system according to claim 2 or 5, it is characterised in that: the filling channel is vertical
The cover board is connected, is equipped with atmospheric equilibrium through-hole in the filling channel side, the atmospheric equilibrium through-hole is located at the absorption
Below the position of filler assembly.
9. chemiluminescence immunoassay system according to claim 6, it is characterised in that: the filling channel vertically connects
The cover board is equipped with atmospheric equilibrium through-hole in the filling channel side, and the atmospheric equilibrium through-hole is located at the absorption with filling out
Below the position for expecting component.
10. chemiluminescence immunoassay system according to claim 7, it is characterised in that: the filling channel vertically connects
The cover board is connect, is equipped with atmospheric equilibrium through-hole in the filling channel side, the atmospheric equilibrium through-hole is located at absorption use
Below the position of filler assembly.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109709349A (en) * | 2019-03-07 | 2019-05-03 | 南京迪格诺斯生物技术有限公司 | A kind of chemiluminescence immunoassay system |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109709349A (en) * | 2019-03-07 | 2019-05-03 | 南京迪格诺斯生物技术有限公司 | A kind of chemiluminescence immunoassay system |
| CN109709349B (en) * | 2019-03-07 | 2024-02-06 | 南京仁迈生物科技有限公司 | Chemiluminescent immunoassay system |
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| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
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| CP01 | Change in the name or title of a patent holder |
Address after: 210000 Hongfeng Science and Technology Park, Nanjing Economic and Technological Development Zone, Jiangsu Province Patentee after: Nanjing renmai Biotechnology Co., Ltd Address before: 210000 Hongfeng Science and Technology Park, Nanjing Economic and Technological Development Zone, Jiangsu Province Patentee before: Nanjing Dignos Biotechnology Co., Ltd. |
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| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of utility model: A chemiluminescence immunoassay system Effective date of registration: 20201228 Granted publication date: 20191119 Pledgee: Bank of China Limited by Share Ltd. Jiangsu branch Pledgor: Nanjing renmai Biotechnology Co.,Ltd. Registration number: Y2020320000272 |
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| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20210601 Granted publication date: 20191119 Pledgee: Bank of China Limited by Share Ltd. Jiangsu branch Pledgor: Nanjing renmai Biotechnology Co.,Ltd. Registration number: Y2020320000272 |