CN107570482A - The removal device and method of the non-specific adsorption thing at interface - Google Patents
The removal device and method of the non-specific adsorption thing at interface Download PDFInfo
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- CN107570482A CN107570482A CN201710548154.8A CN201710548154A CN107570482A CN 107570482 A CN107570482 A CN 107570482A CN 201710548154 A CN201710548154 A CN 201710548154A CN 107570482 A CN107570482 A CN 107570482A
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Abstract
The invention provides a kind of removal device of the non-specific adsorption thing at interface, including：The bulk acoustic resonance device that can be electrically excited, it comprises at least a resonator；One relative plane is set with resonator surface gap certain distance, the gap forms an endless totally enclosed cavity；At least one side of the resonator surface and the plane is modified with detecting element；The cavity is used to be applied in liquid, and the liquid produces the non-specific adsorption thing at least one side that micro-vortex acts on resonator surface and the plane by being driven by the resonator.A kind of minimizing technology is also correspondingly provided.The present invention can be used for the removal to the non-specific adsorption thing of arbitrary interface, have versatility, and have simplicity, miniaturization, compatible with semiconductor technology and the advantages that removal efficiency is high.
Description
Technical field
The present invention relates to sensor technical field, more particularly to a kind of removal device of the non-specific adsorption thing at interface and
Method.
Background technology
Low concentration Protein Detection, such as be directed to biomarker detection, early detection and clinical treatment to cancer have to
Close important effect.Therefore, the sensor based on affinity is widely deployed and studied.These sensors modify detecting element
In sensing surface, the detection of albumen is realized in the interaction by obtaining detecting element and target protein.However, sensors with auxiliary electrode
The problem of all suffering from albumen non-specific adsorption in the detection.The non-specific adsorption of albumen refers to, other in biological sample
Close attached property of the albumen by Van der Waals force, electrostatic force and albumen hydrophobic grouping to substrate is disturbed, non-selective can be deposited on
Sensor or detecting element surface, saltation point 910 as shown in Figure 1, so as to produce very strong background noise, so as to influence target
Detectable limit, sensitivity, selectivity, the repeatability of device or the generation false positive signal of Protein Detection.
Traditional albumen non-specificity minimizing technology is：Apply site closed protein or surface-active in sensor surface
Agent, block site 920 as shown in Figure 1, or make biography in device surface modified polymer or film by chemical modification techniques
Sense interface is hydrophilic or electroneutral is so as to prevent protein adsorption, modifies such film 930 as shown in Figure 1.But these procedures
Complexity, do not possess long-time stability and be likely to be passivated the sensing surface of sensor.
In recent years, some physical methods are also developed and studied the removal for nonspecific proteins.These methods
Principle is to produce interfacial shear force or device volume power using actuator, and this Celik takes albumen non-specific adsorption power, with this
Realize and remove.The generation of the removal power of these methods needs the vibration by device itself to realize, oscillation point as shown in Figure 1
940 by vibrating the method removed, but this physical method for needing the vibration by device itself is only limitted to remove the execution
The nonspecific proteins on device surface, without versatility.
The content of the invention
In view of this, it is a primary object of the present invention to a kind of removal device for the non-specific adsorption thing for providing interface
And method.
A kind of removal device of the non-specific adsorption thing at interface provided by the invention, including：
The bulk acoustic resonance device that can be electrically excited, it comprises at least a resonator；
One relative plane is set with resonator surface gap certain distance, it is endless totally enclosed that the gap forms one
Cavity；
The resonator surface at least one side relative with the plane is modified with detecting element；
The cavity is used to be applied in liquid, and the liquid will be driven generation micro-vortex to act on resonator by the resonator
The non-specific adsorption thing of at least one side of surface and the plane.
By upper, said structure of the present invention, liquid generation micro-vortex in the cavity can be driven to act on by resonator
The non-specific adsorption thing of at least one side of resonator surface and the plane, no longer as background technology is using mode of vibration removal
It is limited to actuator surfaces, with more versatility.
Optionally, the detecting element is modified in the resonator surface；The relatively described resonator surface portion of the plane
It is divided into recess and forms the gap；
By upper, the nonspecific proteins adsorbate on bulk acoustic wave resonator surface can be removed by this way.
Optionally, the plane is modified in the substrate as substrate, the detecting element；The resonator surface with it is described
It is affixed between substrate and is provided with the second plane with hollow-out parts, the hollow-out parts forms the cavity.
, can by this way by upper, the removal to the nonspecific proteins adsorbate of resonator offside, so as to realize
Removal to non-contact face, and can realize that can make removal module contactless, that detachable type is general and other passes accordingly
Sensor is compatible.
Optionally, be additionally provided with for form a mass sensor, electrically connect to examine with the bulk acoustic resonance device
Survey the detection unit of the resonator surface resonance.
By upper, the removal of non-specific adsorption thing selected can be carried out and as matter on same resonator so as to realize
Quantity sensor realizes the sensing detection to albumen, realizes multi-functional.
Optionally, the gap is 50-100 micron distances, and the resonant frequency that the resonator driving produces micro-vortex is
1-10G hertz.
By upper, this clearance distance through experiment with respective resonant frequencies, having preferable removal effect.
Optionally, the resonator of the high frequency bulk acoustic wave resonating device uses array distribution, and/or, resonator has not
Same surface configuration.
By upper, by array distribution, the coverage of device can be increased；And different format surfaces, then it can produce
Fluid mode with a greater variety, different effects is also had for removal.
Optionally, the liquid that applies is the original state difference that the different a variety of solution of viscosity coefficient apply formation successively
Multilayer viscous layer solution.
By upper, using the different multilayer viscous layer solution of the original state for applying formation successively, contribute to non-specific suction
Addendum is peeled off.
Optionally, a variety of solution are two kinds, including PBS, polyoxyethylene (20) sorbierite for diluting 2.7 times
Acid anhydride monolaurate (Tween-20) solution.
By upper, buffer solution uses PBS, does not influence the activity of albumen, and albumen is more stable under buffer solution.
Present invention also offers a kind of minimizing technology based on said apparatus, including step：
A, liquid is applied into the gap of the removal device；
B, the bulk acoustic resonance device of removal device described in electric excitation, its resonator is made to drive the liquid to produce micro-
The non-specific adsorption thing at least one side for acting on resonator surface and the plane that is vortexed.
By upper, the above method of the present invention, liquid generation micro-vortex in the cavity can be driven to act on by resonator
The non-specific adsorption thing of at least one side of resonator surface and the plane, no longer as background technology is using mode of vibration removal
It is limited to actuator surfaces, with more versatility.
Optionally, the electric excitation is controlled, and detects the fluorescence intensity change of different loci albumen, makes caused micro-vortex
Reach maximum when acting on the non-specific adsorption thing for active force.
In summary, the micro-vortex that the present invention excites in a liquid by using high frequency bulk acoustic wave resonator realizes substrate table
Face non-specific adsorption thing removes, and whole system has simplicity, is miniaturized, the spy such as and removal efficiency height compatible with semiconductor technology
Point.
Brief description of the drawings
Fig. 1 is the schematic diagram of nonspecific proteins absorption and existing minimizing technology；
Fig. 2 is the schematic diagram for applying power to bulk acoustic resonance device；
Fig. 3 is the schematic diagram that first embodiment of the invention removes to resonator surface nonspecific proteins adsorbate；
Fig. 4 is the signal that second embodiment of the invention removes to nonspecific proteins adsorbate in resonator offside substrate
Figure；
Fig. 5 is that third embodiment of the invention integrates albumen sensing and the function of non-specific removal in same resonator；
Fig. 6 is detection bulk acoustic resonance device schematic diagram.
Embodiment
Sensor is removed the present invention relates to hydrodynamics method or substrate surface (unless otherwise specified, will in the present invention
Be referred to as interface in the face for adsorbing non-specific adsorption thing) non-specific adsorption thing, the non-specific adsorption thing includes：With various shapes
Protein molecular existing for formula, DNA molecular, the microballoon of nanometer and micron level, cell, the organic matter of micro-nano size, micro-nano size
Inorganic matter etc..Present invention is particularly suitable for the removal of the above-mentioned non-specific adsorption thing of at least micro-nano size.
The present invention mainly uses novel high-frequency bulk acoustic resonance device selective removal interface non-specific adsorption albumen,
After novel high-frequency bulk acoustic resonance device is immersed in into liquid, its be excited caused by acoustic wave energy can dissipate in a liquid production
Raw multiple micro-vortexs, these micro-vortexs produce enough removal power in protein modified substrate and liquid contact surface, so as to realize boundary
The stripping of face non-specific adsorption albumen.
It is in symmetry to detect micro-vortex axial direction both ends, i.e. the removal power at micro-vortex both ends is essentially identical, so as to can be with
The removal of the albumen of the non-specific adsorption in the substrate of resonator offside is realized, that is, is realized and the contactless base of resonator surface
The removal of nonspecific proteins on bottom.So as to, according to the present invention can make removal module contactless, that detachable type is general with
Other sensors are compatible.Detachably refer to that resonator can be used as driver herein, directly act on other sensors surface and enter
Row offside non-specific adsorption thing removes, it is not necessary to and special construction, dismountable benefit refer to resonating device without modifying,
It can be used repeatedly.Also, by nonspecific proteins remove experiment also demonstrate it is above-mentioned to substrate surface, to resonator
The removal of the non-specific adsorption albumen of offside, demonstrate the realizability of the present invention.
Further, since high-frequency reonsator can be used as Mass sen- sitivity sensor, the present invention also provides the device accordingly
Sensing is with removing integrated device.
The above-mentioned selective removal interface non-specific adsorption albumen of the present invention refers to：The nonspecific proteins removal process
In, specific adsorption albumen (detecting with albumen) is not influenceed by removal, and its principle analysis is as follows：Interface non-specific adsorption thing
Remove and be practically due to be vortexed what is realized in drag force caused by interface, the size of this power and the speed positive of micro-vortex
Close, and determine micro-vortex speed is the power applied on resonator, therefore actually this removal power is controllable.Due to spy
The strong adsorption force of the opposite sex is in non-specificity, therefore by above-mentioned controllable mode, can selectivity removal interface it is non-specific
Adhesion protein.Also, also it is able to confirm the removal that this is selective by experiment, the method used in experiment is modified protein battle array
Row, realize specific position and non-specific sites, then fluorescin is applied on site, are found after micro-vortex removes
Only fluorescin disappears on non-specific sites, so as to be confirmed to above-mentioned.
Below, referring to each accompanying drawing of the present invention, in conjunction with specific embodiments, the present invention is further described in detail.
Fig. 2 shows the schematic diagram for applying power to bulk acoustic resonance device, and letter is produced by a radio-frequency signal generator
Number, and bulk acoustic resonance device 110 is supplied to after the signal is amplified by power amplifier, the quilt of bulk acoustic resonance device 110
Electric excitation carries out the conversion of electro-acoustic ripple, and is consumed in the liquid that the acoustic wave energy of caused high frequency sound wave will be submerged in resonator
Dissipate and produce multiple micro-vortexs.Because micro-vortex is in the size of drag force caused by interface and the speed positive correlation of micro-vortex, and determine
Determine micro-vortex speed is on resonator in addition to the power of application, also with the frequency also positive correlation of device, therefore can pass through height
The selection of frequency device and the application of power control drag force.
It should be noted that radiofrequency signal of the present invention can according to demand, using different supply modes, to bulk acoustic wave resonator
Part 110 can apply different performance numbers, can apply different power forms to bulk acoustic resonance device 110, including single-frequency excites
Excited with frequency sweep.According to experimental result, preferable frequency sweep excites device, and frequency sweep excites goes caused by the drag force for producing and changing
Except effect is more uniformly distributed.Also, the high frequency bulk acoustic wave resonating device can use different sizes, different manufacture crafts and
Material, different array distributions and array number, the different surface configurations of array, excite different resonant frequencies.Wherein, lead to
Array distribution is crossed, the coverage of device can be increased；And different format surfaces, then it can produce fluid mould with a greater variety
Formula, different effects is also had for removal.When the resonator in array uses different resonant frequencies, what is inspired is each
Micro-vortex can be different, and caused drag force is also different, i.e., the micro-vortex of different drag forces can be produced on an array, from
And removal effect is more uniformly distributed.
Fig. 3 shows first embodiment of the invention, and the example is the nonspecific proteins suction to bulk acoustic wave resonator surface
Addendum removes, and the non-specific adsorption thing removal device at the interface in the embodiment includes：
The bulk acoustic resonance device 110 that can be electrically excited, it comprises at least a resonator, and it is as micro-vortex driver.
The resonator can be manufactured by MEMS technology；Resonant frequency is preferable in 1-10GHz.In this example, using producible resonance frequency
Rate is the resonator of 2.5G hertz.
Resonator surface is modified with detecting element, and the detecting element is the specific antibody 120 for detecting albumen in this example.
Wherein, (specificity can be modified in these array point positions in resonator surface array modified specificity protein loci
Antibody 120), when carrying out low concentration Protein Detection, being marked with the albumen of fluorescent material both can specifically adsorb in these arrays
Change non-specifically can adsorb at other without on decorating site again on site.
Light enclosed lid 140, cover and set apart from the resonator surface gap, it is endless to be formed one with resonator surface
Totally enclosed cavity.Wherein, through experiment, the height of preferable resonator surface to closing cap 140 is 50 microns to 100 microns
When, removal effect is most strong, is experimentally confirmed, and required solution will also increase if cavity is too high, then effect
Liquid volume increase, its action effect just reduces naturally, and gross energy is conservation；And highly too low lid can be caused to influence
The flowing of fluid, swirl shape can even change.But these situations are different under different capacity, that is to say, that such as
Fruit caving body is very high, then to realize efficient removal effect, then needs to apply higher power to device；And cavity is too low, then may be used
Reduce power.50 microns are highly used described in the present embodiment.Wherein the material of the light enclosed lid 140 can be cover glass or silicon
Piece etc., when the purpose using lightweight surfaces is to form the cavity of approximate closing, avoid damaging cavity, also avoid overweight making chamber
Body height is inaccurate.The plane layer with hollow out can be pasted between light enclosed lid 140 and resonator surface, the plane layer can
For 50-100 micron thickness, so as to which the hollow part forms above-mentioned cavity.
By it is transparent it is pressure sensitive adhesive double coated exemplified by illustrate the cavity constituted mode：Pressure sensitive adhesive double coated thickness is about 50 microns, can be pasted
Resonator surface, it corresponds to the position hollow out of resonator surface, and then the double faced adhesive tape opposite side pastes the light enclosed lid
140, so as to which the hollow out position forms the cavity.It is described pressure sensitive adhesive double coated to use transparent adhesive tape or photoresist, PDMS materials
Deng replacement.
The endless totally enclosed cavity formed is used to store liquid, and the liquid will be driven by the resonator and produce institute
State micro-vortex and act on the non-specific adsorption thing such as non-specific adsorption albumen of resonator surface.The liquid is molten except pure water
Outside liquid, following at least one the formed solution for contributing to non-specific adsorption thing to peel off can also be：There can be surface clear
The solution of clean effect, such as surface-active agents；The solution removed by frictional property, such as reagent containing micro-nano particle；Can be with
The biomolecule of remover (albumen 130 of the i.e. described non-specific adsorption on device surface or detecting element surface) interaction
Reagent, such as organic solution containing cleaning properties.Preferably, select the different solution of two kinds of viscosity coefficients not blended gradually
Apply, to form two layers different of viscous layer solution of original state, have selected in this example a kind of water miscible surfactant with
Buffer solution (is referred to described in later-mentioned step S1).
Albumen 130 of the non-specific adsorption on device surface or detecting element surface is also schematically illustrated in Fig. 3, with
And the protein 15 0 (i.e. the detection object of detecting element) with detecting element specific binding.
Below to the operation principle of the embodiment, i.e., the nonspecific proteins adsorbate on bulk acoustic wave resonator surface is removed
Principle is further described below, including step：
S1：By pipettor, apply 5 microlitres of buffer solution and the polyoxyethylene of 5 microlitres of 2.7 times of dilutions successively in cavity
(20) sorbitanmonolaureate (Tween-20) the solution cement layer different as two layers of dynamic viscosity.Wherein, buffer solution
Preferably PBS, in order to do not influence the activity of albumen, albumen is more stable under buffer solution.
S2：When applying under frequency sweep mode 500 milliwatt power to resonator, by instrument, such as high speed microscope, it can detect
It is leak into acoustic wave energy in liquid and produces micro-vortex, now, two layers different of cement layer of initial viscosity is under the disturbance of vortex
Produce special exercise and mutual phased soln, be now vortexed and reach maximum in drag force caused by resonator surface, its magnitude with it is non-
Specific proteins absorption is identical in the viscous force of resonator surface, thus realizes the removal of resonator surface nonspecific proteins.
Wherein, the adjustment of power can be by the fluorescence intensity change of different loci albumen before and after judging that power applies, to device
Part removal efficiency is evaluated, to be finally set in best power.In addition, fully, uniformly being removed to realize, what can be selected applies
It is 10 minutes to add Power x Time.
As Fig. 4 shows the second embodiment of the present invention, this provides the nonspecific proteins for removing resonator offside
The method of adsorbate, that is, realize the removal with the nonspecific proteins in the non-contact substrate of resonator.For simplify describe Fig. 4 in
Fig. 3 identicals part represents that following same parts also simplify description with identical label, and Fig. 4 shown devices include：
The bulk acoustic resonance device 110 that can be electrically excited, it comprises at least a resonator, the driver as vortex.
The side of the resonator surface face, gap certain distance are arranged to albumen substrate 160, are modified with detection thereon
Element, can array modified specificity protein loci.Wherein, the clearance distance of resonator surface to albumen substrate 160 is micro- for 50
Rice is about 50 microns to 100 microns, in this example.The gap can be applied in liquid.Albumen substrate 160 and the resonator surface structure
Into endless totally enclosed cavity, this cavity can be integrally fixed in resonator surface.This cavity can additionally be added
A cavity, material can be pressure sensitive adhesive double coated, and PDMS photoresists etc. are formed, such as can be have hollow-out parts pressure sensitive adhesive double coated, one
Face sticks in resonator surface, simultaneously sticks in the albumen substrate 160, and hollow-out parts face resonant structure forms the cavity.
The difference of the embodiment and first embodiment is, the incomplete closing cap of lightweight in first embodiment is replaced by
The substrate of protein arrays is modified with, and resonator surface is not modified, this is to be based on detecting high frequency bulk acoustic wave resonator in liquid
In the micro-vortex that excites have at a high speed, the feature of upper and lower form near symmetrical, micro-vortex has for the upper and lower surface being in contact with it
There is approximate action effect.
As Fig. 5 shows the third embodiment of the present invention, relative to first embodiment, the bulk acoustic wave being electrically excited
Resonating device 110 can also be detected by the Network Analyzer shown in Fig. 6 simultaneously, can be chosen on same resonator so as to realize
The carry out non-specific adsorption thing removal selected and the sensing detection as mass sensor realization to albumen.
This is to be based on, because high frequency bulk acoustic wave resonator can be as highly sensitive mass sensor (when by materials such as albumen
After absorption, the change of its quality causes the change of harmonic wave and is detected), therefore, while shown in connected graph 6 during circuit, work as conduct
Sensor is in use, the change of the resonant frequency of resonator can be monitored by Static Detection mode so as to obtain protein modified and egg
The information sensed in vain；Device only shown in connected graph 2 during circuit, shown by composition first embodiment, can be to resonator surface
Nonspecific proteins adsorbate be removed.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Within refreshing and principle, any modification, equivalent substitution and improvements made etc., for example, above-mentioned light enclosed lid and resonator surface structure
Into endless totally enclosed cavity in solution applying mode, except above two solution it is not blended gradually apply in addition to, may be used also
Gradually apply outer to form the different multilayer viscous layer of original state to be that a variety of solution are not blended, or single solution is direct
Apply or different solutions mixing is after-applied, these should be included in the scope of the protection.
Claims (10)
- A kind of 1. removal device of the non-specific adsorption thing at interface, it is characterised in that including：The bulk acoustic resonance device that can be electrically excited, it comprises at least a resonator；One relative plane is set with resonator surface gap certain distance, the gap forms an endless totally enclosed chamber Body；At least one side of the resonator surface and the plane is modified with detecting element；The cavity is used to be applied in liquid, and the liquid will be driven generation micro-vortex to act on resonator surface by the resonator With the non-specific adsorption thing of at least one side of the plane.
- 2. device according to claim 1, it is characterised in thatThe detecting element is modified in the resonator surface；The relatively described resonator surface part of the plane is that recess forms the cavity.
- 3. device according to claim 1, it is characterised in thatThe plane is modified in the substrate as substrate, the detecting element；It is affixed between the resonator surface and the substrate and is provided with the second plane with hollow-out parts, the hollow-out parts forms institute State cavity.
- 4. device according to any one of claims 1 to 3, it is characterised in that be additionally provided with for forming a mass sensor , electrically connect to detect the detection unit of the resonator surface resonance with the bulk acoustic resonance device.
- 5. device according to any one of claims 1 to 3, it is characterised in that the gap is 50-100 micron distances, described The resonant frequency that resonator driving produces micro-vortex is 1-10G hertz.
- 6. device according to any one of claims 1 to 3, it is characterised in thatThe resonator of the high frequency bulk acoustic wave resonating device uses array distribution, and/or, resonator has different surface shapes Shape.
- 7. device according to claim 1, it is characterised in that the liquid that applies is the different a variety of solution of viscosity coefficient Apply the different multilayer viscous layer solution of the original state of formation successively.
- 8. device according to claim 7, it is characterised in that a variety of solution are two kinds, including PBS, dilute Release 2.7 times polyoxyethylene (20) sorbitanmonolaureate (Tween-20) solution.
- 9. a kind of minimizing technology based on any described device of claim 1 to 8, it is characterised in that including step：A, liquid is applied into the gap of the removal device；B, the bulk acoustic resonance device of removal device described in electric excitation, its resonator is made to drive the liquid to produce micro-vortex Act on the non-specific adsorption thing of at least one side of resonator surface and the plane.
- 10. according to the method for claim 9, it is characterised in thatThe electric excitation is controlled, and detects the fluorescence intensity change of different loci albumen, acts on caused micro-vortex described Reach maximum during non-specific adsorption thing for active force.
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Application publication date: 20180112 |