CN106104271A - There is micro-fluid chip and the manufacture thereof in conical bead trapping chamber - Google Patents
There is micro-fluid chip and the manufacture thereof in conical bead trapping chamber Download PDFInfo
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- CN106104271A CN106104271A CN201580012689.2A CN201580012689A CN106104271A CN 106104271 A CN106104271 A CN 106104271A CN 201580012689 A CN201580012689 A CN 201580012689A CN 106104271 A CN106104271 A CN 106104271A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0668—Trapping microscopic beads
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0887—Laminated structure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/08—Regulating or influencing the flow resistance
- B01L2400/084—Passive control of flow resistance
- B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/554—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being a biological cell or cell fragment, e.g. bacteria, yeast cells
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
- H01L21/00—Processes or apparatus adapted for the manufacture or treatment of semiconductor or solid state devices or of parts thereof
Abstract
A kind of micro-fluid chip (100), including layer (10,60), the array (30) in pearl trapping chamber (20) is located in described layer (10,60), each of wherein said chamber (20) has the taper limited by one or more sidewalls (21 24), described sidewall is all hydrophilic, and each in wherein said chamber (20) extends in the thickness of described layer (10,60) as blind hole.Also provide for the manufacture method of such chip.
Description
Technical field
The present invention relates generally to the field of micro-fluid chip, and the most such as applies for bioanalysis
, equipped with the micro-fluid chip in pearl trapping chamber, and relative manufacturing process.
Background technology
Microfluidic device is generally referred to as micro-manufacture device, and it is used for pumping, samples, mixes, analyzes and quantitatively to liquor charging
Body.The prominent features of microfluidic device is derived from the special behavior that liquid shows on micrometer length scales.Liquid in microfluidic device
The flowing of body can be typically stratiform.Volume far below one nanoliter can have the lateral chi in micrometer range by manufacture
Very little structure obtains.Therefore the reaction being restricted (by the diffusion of reactant) in large scale can be accelerated.Microfluid
Device thus be accordingly used in various application.
Many microfluidic devices have user's chip interface and Guan Bi flow path.Guan Bi flow path is easy to Functional Unit
Part (such as, heater, blender, pump, UV-detector, valve etc.) is incorporated in a device, minimize simultaneously and leak and
The problem that evaporation is relevant.
Generally, the receptor on surface is for combining the specific analyte needing in the sample to be detected.After the coupling, sample
Product and interfering material can be washed off.Then can be directly (such as via the change of quality, refractive index etc.) or indirectly (glimmering
Light immunoassay etc.) detection receptor-analyte complex.Although microfluidic device is the promising device for analyzing, but
It is that receptor is incorporated at present the inside of micro-fluid chip is very challenging.
Having pointed out solution, wherein receptor is patterned on the surface of micro-fluid chip.In more detail, microfluid core
Sheet can seal with the PDMS layer being patterned with capturing the line of antibody on it.In this case, by using masterplate from molten
Liquid absorption antibody realizes patterning capture antibody.But, such method is troublesome, has poor efficiency, needs masterplate, and
And it is slower additionally, due to adsorption process.It addition, PDMS is expensive and contaminated surface (～20 minute contact time after
Make the water-wetted surface hydrophobic).
Other solution known relies on the microballon for measuring for a long time.Here, pearl is typically coated with being subject to
Body.Pearl can in the solution (such as, magnetic bead, or the single pearl in capillary tube) or depositing/be positioned at the spy of micro-fluid chip
Used after determining in region.Two kinds of situations can be distinguished:
Magnetic bead: with rinsing, pearl is separated with sample from interfering material by using magnet.But magnetic bead is than non-magnetic bead more
Expensive and be difficult to prepare.And, these pearls are opaque and are not to be suitable for very much optics/mensuration based on fluorescence；With
And
Non-magnetic bead, has been for the many technology of non-magnetic bead exploitation for positioning/handle pearl in microfluidic device.
But, such technology has following defect.They need:
-particular activated device (electrode, magnetic texure focus on light, transducer, piezoelectric structure etc.), and therefore complicated and
Expensive；Or
The geometry in particular (certain radius of curvature, contraction flow region) in-microfluidic flow path, it should be noted that with and without
Pearl, the flowed friction of chip can be different significantly, and the constancy system of the pearl being trapped be a problem.And, specific
The viscosity of sample/liquid can be problem.
As it has been described above, some solutions use contraction flow region or " filter ", it is directly the flowing in micro-fluid chip
The part in path, for trapping pearl.But, such solution causes the balance between signal intensity and signal quality.
Pearl stability is a problem further.
The present invention is proposed for the determination solution of pearl stability problem.
Summary of the invention
According to first aspect, the present invention is embodied as a kind of micro-fluid chip, and it includes that layer, the array in pearl trapping chamber set
In said layer, each in wherein said chamber has the taper limited by one or more sidewalls, and described sidewall is all parent
Water, and each of wherein said chamber extends in the thickness of described layer as blind hole.
In an embodiment, at least some in described chamber has the pyramid shape formed by sidewall, and described sidewall is all parent
Water, and wherein it is preferred to, this pyramid shape is substantially limited by least four sidewall.
Preferably, described layer includes one or more semiconductor elements of such as silicon, and the pyramid shape tool in described chamber
There is the geometry consistent with the anisotropic etching process of the manufacture in the described chamber in described layer.
In a preferred embodiment, at least one in described chamber, preferred major part are all filled with pearl, preferably microballon, and it is put down
Be respectively provided with between 1 to 40 μm, diameter between preferably 2 to 20 μm and between more preferably 2 to 10 μm.
Preferably, the major part in the chamber of described array is all filled with only one pearl, and it preferably averagely has 1 and arrives
The microballon of the diameter between 40 μm, between preferably 2 to 20 μm and between more preferably 2 to 10 μm.
In an embodiment, the average-size of the opening in described chamber and the ratio of the average diameter of the pearl in described chamber are 1.0
Between 2.4, and preferably between 1.4 to 2.0.
Preferably, the mean depth in described chamber is at least 0.5, preferably with the ratio of the average diameter of the pearl in described chamber
1.0, and more preferably 1.3.
In a preferred embodiment, two or more subgroups of at least one in described chamber are by least one microchannel even
Connecing, described subgroup is preferably the row or column of the array in described chamber, and wherein, it is highly preferred that in described subgroup one or
Multiple being defined in channel portion, diapire or the roof of described channel portion are formed by the surface of described layer.
Preferably, described array is sealed by the cap rock extended relative to the array in described chamber.
In an embodiment, described chip includes some arrays in one or more pearl trapping chamber, and wherein said array is preferred
Be inserted between different pairs of channel portion.
Preferably, described chip includes being correspondingly situated in one or more chambeies of at least two in described some arrays
The dissimilar pearl of at least two, wherein said different types of pearl is preferably in terms of size, coating, material and/or color
Different.
According on the other hand, the present invention is embodied as a kind of according to any one microfluid core in above example
The manufacture method of sheet, described method includes:
The micro-fluid chip main body with layer is provided；And
Manufacturing the array in pearl trapping chamber in said layer, each in wherein said chamber has by one or more sidewalls
Limit taper, described sidewall is all hydrophilic, and each of wherein said chamber as blind hole in the thickness of described layer
Extend.
Preferably, manufacture described array and include that being preferably used the anisotropic etching process from limit is etched anisotropically through
Described layer is to obtain described chamber.
In an embodiment, described method also includes: pearl is deposited to the institute of described array by the drop of distribution pearl solution
State in chamber；And with being positioned to the cap rock described array of sealing relative to the extension of the array in described chamber, and wherein seal described
Array preferably includes the described cap rock of lamination.
Preferably, deposition pearl after and before sealing, described method the most also includes: be dried at it
In there is the array in described chamber of pearl；Preferably by rinsing described array with rinse solution and/or being used for adhering to by applying
The excessive pearl of the inside not being trapped in described chamber removed by the band of excess pearl；And if desired, after drying has pearl wherein
The array in described chamber.
Now will be by non-limitative example and be described with reference to the drawings and be embodied as apparatus and method of the present invention.In figure
Shown technical characteristic is not necessarily drawn to.
Accompanying drawing explanation
Fig. 1, Fig. 2, Figure 12 and Figure 13 all show that the simplification of the micro-fluid chip of different embodiment according to the subject invention represents
2D (bowing) view；
Fig. 3 shows: the sectional view (upper figure) simplifying expression of the pearl being trapped in pyramid chamber related in an embodiment
With top view (figure below)；
Fig. 4 is the scanning electron microscopy picture of the pearl being trapped in pyramid chamber related in an embodiment；
Fig. 5 A-5I schematically shows the detailed step of the manufacture method of micro-fluid chip according to an embodiment of the invention
Suddenly；
Fig. 6 schematically shows the selected step of the manufacture method of Fig. 5, the wherein cross section of upper row's display micro-fluid chip
Figure, and lower row shows corresponding top view；
Similarly, Fig. 7-9 schematically shows the selected step of modification of manufacture method of Fig. 5；
Figure 10 corresponds to the flow chart of Fig. 5；
Figure 11 shows the 2D view simplifying expression in the pyramid chamber of the improvement related in an embodiment.Leftmost diagram is to bow
View, and other figure is sectional view；
Figure 14 shows the egative film of the fluoroscopic image of the pearl in the array being trapped in the chamber according to embodiments of the invention acquisition；
And
Figure 15 is the sectional view simplifying expression of the pearl being trapped in frustoconical chamber related in an embodiment.
Detailed description of the invention
First, and generally with reference to Fig. 1-4 and 11-13, the aspect of the present invention relating to micro-fluid chip 100 is described.
Chip includes working lining 10,60 significantly, and described layer includes the array 30 being located at pearl trapping chamber 20 (also referred to as trap) therein.
It is apparent that each in chamber 20 has the taper surrounded by one or more sidewall 21-24.Importantly, this or these side
Wall is hydrophilic and each in chamber 20 extends in the thickness of working lining 10,60 as blind hole.
Working lining 10,60 can be substrate layer 10 or cap rock 60.Chamber is preferably disposed in the thickness of layer, the surface of described layer
It is microfluidic channel or the diapire of channel part 12a or roof, as shown in Figure 2.
In this application, " taper " also illustrates that " taper ", i.e. has the form of similar cone.By " taper ", and with
One general definition of cone as one man (http://en.wikipedia.org/wiki/Cone), it represents from planar base portion
(corresponding to the opening 28 in chamber) smoothly taper to have the area of section less than base portion 28 opposed end (such as, summit or
Truncation surface) 3 dimensional coil geometry, described planar base portion is not necessarily circle.Other definition assumes that cone is the specific feelings of pyramid
Condition, cone will be defined as the pyramid with circular cross section, for example, see http://mathworld.wolfram.com/
Cone.html.It is assumed herein that taper includes the pyramid (having polygonal base) as particular case.It is to say, it is current
Taper need not have circular cross section.
Limit owing to manufacturing, taper however may be truncated, or the most do not terminate as perfection, punctiform vertex.It practice,
Even can utilize frustum, as described later.It is further illustrated as described in immediately above and as in the following examples,
Bellmouth will preferably have polygonal base (base portion is corresponding to the perforate in chamber 20), and therefore (have polygon for pyramid
Shape base portion).Taper can be referred to as bevel taper shape in this case, and reason is that pearl 50 (may pass through from directly spreading
Liquid dissemination technology) drop on the top of the lower array 30 of pearl trap is captured, and wherein trap is chamber 20, and it at top is
Open wide, i.e. inwardly village hollowing.But, this configuration can overturn, thus has the chamber being located in cover cap, sees Fig. 7.
Also other solution tested compared to the present inventor, such as, use post or the side in through hole shape chamber for trap
Case, uses blind hole shape conical cavity result to prove in terms of the stability of pearl for pearl trap the most favourable.It is, by
This trap produced demonstrates the unprecedented tendency keeping pearl 50 after being flushed.Embodiments of the invention are applied to microballon
(such as, having the pearl of receptor in its surface, wherein according to the Normal practice in bioassay, such receptor may be used for
Part in binding soln), there is the complexity of appropriateness and need not additional means (such as vacuum, magnetic field or electric field) by pearl guarantor
Hold in place.
The cause significantly improved of the stability of pearl is not clear；The stability improved is likely due to when pearl is trapped in chamber
In, time i.e. at the tip of bellmouth, laterally (conical cavity preferably protects pearl to avoid fluid (gas or liquid) when rinsing
Lateral, the impact of move in plane) and/or below pearl occur interface and/or mechanical phenomenon.And, (the one of conical cavity
Individual or multiple) sidewall needs to be hydrophilic, in order to its work.
Current design and corresponding manufacture method also allow for placing pearl before covering trap, with most (even if not being
Solution known to all) is different.
At manufacture view, the present inventor has been noted that after scattered tear drop solution and before rinsing excess pearl, to pearl
50 and trap 20 be dried the stability the most substantially improving the pearl being trapped.This will be described the most in detail.
Known to some prior art solution (for example, see, Sohn et al., biosensor and bioelectronics device
Part, volume 21,2005, the 303-312 page) in, through hole shape pyramid chamber is manufactured, in order to trapping " huge " pearl, i.e. directly
The pearl of at least one magnitude bigger than pearl contemplated herein on footpath.But, owing to the pyramid chamber manufactured is logical in this case
Hole, they necessarily have minimum-depth the biggest for the application.And, such solution need pickup-
Place tool and the applying of dorsal part vacuum, it may not apply to less pearl, microballon as contemplated herein, microballon typically tool
There is the polymeric beads (such pearl typically obtains) of the diameter in the range of 1-40 μm from liquid suspension.
Although the present invention allows to obtain stable pearl in chamber 20, but it is not precluded from the additional of dielectric DEP electrode etc.
Use.DEP electrode can be physically located in trap or near, such as make pearl 50 float, thus such as reclaim pearl (the need to
If), or pearl is more exposed in the microchannel residing for the array 30 of trap the solution of flowing.It addition, in embodiment
In, the surface in chamber can such as use Al to metallize will pass through reflection to amplify fluorescence signal.
Referring more especially to now Fig. 3, Fig. 4, Figure 11 and Figure 15, some or all in chamber 20 can be endowed pyramid
Shape, i.e. has the taper volume of polygonal base, it is, the chamber formed by some, different lateral 21-24, described wall is being worked as
It is all hydrophilic in the case of before.Pyramid shape can be such as tetrahedron, quadrangular pyramid or more complicated.Although preferably, this shape
Substantially limited by (at least) four sidewall 21-24.Therefore, such chamber 20 typically has triangular wall 21-24, and it is preferable
Converge to a little, but more likely converge to truncation surface 29.The planar base portion of the opening 28 limiting chamber has polygon, and it is at least
It is three limits, and preferably four limits.In the case of the latter, pyramid shape is substantially by least four sidewall 21-24
Limit.
It should be noted that, wall needs not be perfect flat wall, it may be possible to due to the danger in manufacturing process.But, it should note
Meaning, preferably fabricated technology (see below) disclosed herein is essentially prevented from this situation.Such as due to photoetching process, some are residual
Surplus and deficiency falls into and there may be, but this is actually rare.On the contrary, sidewall is typically smooth, reason is anisotropy erosion
Quarter follows crystal face.Such as can control shape as shown in Figure 11 by amendment mask (layout), so final form is at least
Can be controlled and predictable in an embodiment.One or more in sidewall such as can be structured or shape, as
Shown in Figure 11, in its mesospore is structured 23a-d, thus allows localized liquid to inject, and pearl discharges and attracts,
For not using those embodiments of cap rock.
Compared to the chamber of " more " cone, in terms of the stability of pearl, it has been found that use pyramid shape to cause better
Result.Such reason is not also entirely understood.This is likely due at the free space stayed in the turning of spherical bead,
This allows residual liquid evaporation, sees Fig. 3, different from cone shape.
Fig. 4 is the scanning electron microscopy picture showing the pearl being trapped in pyramid chamber.In the images:
-" EHT " represents the electronic high voltage in terms of kilovolt kV；
-" WD " represents the operating distance between sample surfaces and the lower part of lens；
-" Mag " is amplification；
-" inclination angle " represents the normal angle relative to the axis of electron gun of sample stage；And
-" signal A=SE2 " instruction uses the detector of secondary electron.
With reference to Fig. 4: in an embodiment, working lining 10 includes one or more semiconductor elements of such as silicon, and chamber 20
Pyramid shape have with for obtaining the geometry that the manufacturing process in chamber is consistent, described process advan ground use anisotropy
(typically using the silicon chip with<100>crystal orientation, reason is other orientation to etch process, and such as<111>will not produce desired
Pyramid shape).Now, although silicon is preferred, but other quasiconductor that is applicable to considered below, such as Group IV element, example
Such as Ge or compound semiconductor, such as SiGe, other compound III-V or II-VI material, and their corresponding oxide
Or nitride.For example, it is possible to be etched anisotropically through GaAs and Ge.And, this method the principle on which can apply to one
A little metal levels and corresponding oxide.But, metal level is the most practical.Especially, they may not have phase on layer thickness
When crystal homogeneity.Furthermore, it is possible to such as expection uses Al2O3 surface.Al2O3 can serve as reaching the thin of 100-200nm
Film electrolyte, and by sputtering or deposited by ald ALD.The latter is expensive but that quality is high technology.And,
The thickness of the layer needed for microballon trapping has challenge for using ALD.
Pyramid shape is unexpectedly compatible with anisotropic etching process.For the semiconductor element of Si or similar, manufacture chamber
Anisotropic etching process cause the desirable angle of between planar base portion (that is, opening 28) and the adjacent wall of pyramid 54.7 °, this
Determine again the degree of depth-width ratio of pyramid.It can be seen in fig. 4 that such technique allows easily manufacturing of chamber with cleaning.So
And, this desirable angle is likely to be due to little defect and changes slightly.
In modification, it is possible to use dry film photoresist and more generally, it is contemplated that some polymer, plastics and gold
Belong to.Preferably, using epoxy radicals dry film photoresist, it can be patterned by hot padding.Hot padding can also be applied to it
Its plastic material (such as, PMMA, COC, Merlon), the plastics patterning method common for described material may be not suitable for
In producing pyramid microcavity.Such as, after the anisotropic etching using Si manufactures the array in chamber, can be electroplated by Ni
Fill chamber.Then electroplated Ni layer departs from, and is formed the mould used in thermal imprint process subsequently or die.Can pass through
Pressure and temperature is applied to the die that contacts with plastics and then cooling and the demoulding and on plastic layer generation Si chamber
True copy thing.
Referring now more particularly to Figure 14, in an embodiment, preparation chip makes its chamber (that is, at least in chamber 20
A bit, preferably most of) all it is filled with pearl 50,51,52.As it was earlier mentioned, such pearl is preferably microballon, have the most on average
There is the diameter between 1 to 40 μm.For application as contemplated herein, between 2 to 20 μm and between more preferably 2 to 10 μm
Diameter be preferred.Pearl is preferably polymeric beads, such as polystyrene bead, but can also use in principle silica beads or
Latex bead.
As it is shown in the figures, the size of the cross-sectional area in chamber is equivalent to the size of pearl.Pearl trapping chamber is to be designed specifically to
The trap of the single pearl of trapping.Therefore, pearl can even stick to the point in chamber typically two (or more) point cantact cavity wall
At end." chamber " is expected in this article and is different from the object of microchannel or other microfluidic features.Due to solution presented herein
Certainly scheme, the great majority in the chamber 20 in array 30 all can be filled with only one pearl 50,51,52, as seen in Figure 14.
At present, be given about the chamber details to the relative size of pearl, its stability allowing to optimize the pearl being trapped and falling into
The occupancy of trap.Refer more especially to Fig. 3,11 and 15: average (linearly) size of chamber opening 28 and the ratio of average bead diameter
Should be preferably between 1.0 to 2.4.And, optimum is had observed that (in occupancy for the ratio between 1.4 to 2.4
Aspect) (it is, average 40-60% occupancy).When ratio is in 1.4-2.0, these results are improved further (in flushing
Afterwards it is observed that averagely reach 60% occupancy).In some cases, it is possible to obtain reach the occupancy of 90%.And,
Above ratio is maintained at less than 2.0 or preferably less than 1.8 allow to be essentially prevented from the multiple of same chamber and take.
Such as, and the various results collected according to inventor, for 10 μm (diameter) pearl, limit the opening 28 in chamber 20
The full-size of planar base portion should be preferably less than 24 μm, and even more preferably less than 18 μm.These digit preferences such as by
Fluoroscopic image shown in Figure 14 confirms.The latter actually shows what the method being trapped according to reference Fig. 5 description subsequently obtained
The egative film of the fluoroscopic image of the pearl in the array in chamber.The TMAH etching deep actually by 13 μm performing Si substrate obtains chip
Chamber；Use 200 μ m 500 μm arrays.By by 10 μm Fluoro-MAX of about 200nL (from Thermo
ScientificTM) pearl solution (undiluted) is pipetted on each array and integrates pearl.Therefore, in each of image of Figure 14
In, use the pearl of 10 μ m diameter.The lateral dimensions (the most invisible) of the opening in chamber from 8 μm to 24 μm, and corresponding
Last occupancy rise to 63% from 5%, 18-20 μm perforate is seen to the peak value of 80%-90%.For these specific feelings
Condition, it is thus achieved that actual percentage be: 8 μm: 5.4%；10 μm: 24.4%；12 μm: 32.5%；14 μm: 46.3%；16 μm:
51.6%；18 μm: 80.1%；20 μm: 89.9%；22 μm: 72.2%；And 24 μm: 63.2%.But, multiple take appearance
Increase in 18 μm and on this.Therefore, therefore the scope of the size of perforate can be limited to (for 10 μ m diameter pearls)
Such as 14-18 μm, even 16-18 μm.As one man, the average-size of chamber opening 28 can be limited with the ratio of average bead diameter
To 1.4-1.8 or 1.6-1.8.It should be noted that the percent value caused by above particular case be different from previously mentioned averagely
Value, reason is that the latter collects from various experiences.
With reference now to Fig. 3 and Figure 15: the mean depth in chamber 20 is preferably at least with the ratio of the average diameter of pearl 50
0.5, more preferably 1.0, and it is more preferably 1.3.Such as, the present inventor the experiment carried out shows 10 μ m diameter
Pearl can keep stable in 8 μ m 8 μm openings, but finds that the last occupancy/stability of pearl is not good enough in this case
's.When using from the anisotropic etching process of limit, the degree of depth of such size meaning about 5 μm.Therefore, the average cavity degree of depth
Can as little as 0.5 with the ratio of average bead diameter.Now, for the cone through anisotropic etching, wrap completely when hope obtains
When burying (that is, embedding) pearl in chamber, the degree of depth of at least (about) 1.0d (wherein d is the diameter of pearl) is necessary, it is assumed that taper
Chamber is truncated and truncation surface 29 is sufficiently large to accommodate pearl, as shown in Figure 15.Such as, 10 μm pearls can be fully embedded to chamber
In, if opening is at least 19.33 μ m 19.33 μm, and minimum at least 10 μm of the degree of depth, base etching during use；Time base
Etching stopped etching before apex merges in all planes.As fruit caving is not truncated, (they have punctiform vertex, i.e. certainly
Limit etching), then ratio needs at least about 1.3, as shown in Figure 3.
And, inventors have also recognized that pearl is without the need for being fully embedded in chamber.It is to say, a part for pearl is permissible
(that is, prominent) occurs above the plane of opening 28 to keep the stability of satisfaction, as shown in Figure 5 (not in scale).Therefore
Can correspondingly reduce above-mentioned ratio.
With reference now to Fig. 1, Fig. 2, Fig. 6-9 and Figure 12-13, in an embodiment, micro-fluid chip 100 can have chamber 20
Row or column in two or more subgroups 32, such as array 30, two of which subgroup is connected by least one microchannel 14.Excellent
Selection of land, one or more in these subgroups are defined in channel portion 12a, the diapire of described channel portion or roof by
Working lining 10 limits.Such configuration allows multiplexing, prevents the cross-contamination between subgroup 32 simultaneously.It is preferred, therefore, that
It is the subgroup 32 that array 30 is divided into pearl 50, in order to be partially isolating subgroup.The array in chamber such as can be arranged to mate trace
Titration hole canonical coordinates: on microtitration plate, hole with 9,4.5 or 2.25mm positioned at intervals on secondary dot matrix.This can
So that using robotic to distribute the solution of pearl.In order to the increase of the array in chamber is integrated, it is also possible to select hexagonal-lattice.
Such as, in the experiment shown in Figure 14 use chip design include array, its along snakelike microfluidic channel (200 μm width, 14
μm is deep) arrange distribution with hexagonal-lattice, wherein it is about 1.1mm from an array to another angle diagonal distance.This layout
Allow the gross area of 3mm × 5mm in 10 single arrays of placement (each take 200 μ m 500 μm areas), and allow
By liquid droplet distribution nonjoinder drop or cross-contamination to each array of 200nL pearl solution.
Then, seen in " finally " device as shown in figures 5-9, the array 30 in the chamber 20 of micro-fluid chip 100 is preferably
Sealed by the cap rock 60 extended relative to array 30.Cap rock protection and annular seal space and their content.In addition to chamber 20,
Cap rock 60 will seal other microfluidic structures being typically found on chip 100.The example of such micro structure is: load
Pad 11, detection antibody (or dAb) deposition region 70, capillary pump 16 or passage 18, as shown in figs. 1 and 2.
Although it should be noted that, cap rock 60 extends relative to array 30, but it need not contact chamber, reason be typically require between
Gap is to form microfluidic channel.In the manufacturing process of Fig. 9, chamber is sealed by cap rock, and reason is that microchannel is arranged between chamber.And
And, in other described in this article manufacturing technology, epiphragma 60 is actually not directly contacted with chamber, always has and is deposited by channel layer
(Fig. 6) or etching (Fig. 7 and the 8) gap that introduces.Typically, channel depth is between 1 to 20 μm.More shallow passage will produce more
High flowed friction and less thickness evenness, and the manufacture of more deep channel will be more difficult and/or time-consuming.It is said that in general, can
The minimum lateral size of the feature to manufacture increases along with the degree of depth of passage owing to the aspect ratio of passage manufacturing technology limits.Mesh
Front manufacturing technology (wet etching or dry ecthing or dry film photoresist patterning) has the aspect ratio higher than 1 by easily providing
Structure (feature that 20 μm in the passage that such as, 20 μm are deep are wide) is without substantial amounts of parameter optimization effort.
Thus, channel height (or degree of depth) can be adjusted to assure that pearl will not during flow depart from.Due to this
Solution, the stability of pearl significantly improves.And, may be always in the case of the flowing of high current body is present in microchannel
There is the residual risk of more loosening pearls.Now, if the microfluidic channel degree of depth is slightly less than diameter (such as, the example previously of pearl
Less than 10 μm in son), pearl during flow will never depart from.But, the least channel depth (such as, 1 μm) will increase hydraulic pressure
Resistance and reduce total liquid capacity (volume) of chip；Increase is loosened some pearls by the biggest degree of depth (such as, more than 10 μm)
Probability.Remove the situation of excess pearl using to bring (that is, the pearl in the inside in chamber will retain and in the excess of channel interior
Band will be adhered to) under to have the channel depth of the diameter being slightly less than pearl will be also useful.
Referring now more particularly to Figure 12 and Figure 13, chip 100 can actually include some array 30 (each array bags
Include one or more pearl trapping chamber 20).These arrays 30 are preferably inserted in different pairs of microchannel in specifying flow path
Between part, the most in parallel or series.Such as, these channel parts can be made up of parallel split tunnel, in Figure 13 significantly
Shown in, thus avoid from a part to another cross-contamination, or can (in Figure 12) arranged in series.They are also
Can be arranged in the channel part of serpentine channel (not shown).
Then, it is also possible to favorably use the different types of pearl 51 in the chamber 20 being positioned at (one or more) array 30,
52.Due to preferred deposition technique, pearl 51,52 is typically deposited in different array 30.It is, for example possible to use two kinds dissimilar
Pearl: a type of pearl 51 is for detection of analytes, and another type of pearl 52 is used for compareing, such as Figure 12, as shown in 13.As
Previously described, depend on the actual application sought, array 30 can be with serial or parallel connection.Intended most application (example in this article
As, bioanalysis) in, pearl is different in terms of coating.But pearl can be more generally in size, coating, material and/or color side
Face is different.
Sealed before loading pearl (flowing via pearl solution) it should be noted that, the solution of prior art focuses principally on
Device.This makes it difficult to (if possible) and has the different types of pearl in the precalculated position being positioned in device.Therefore, this is right
More step is actually meant that in bioanalysis.
According on the other hand, the present invention can be specifically embodied as the manufacture method of chip 100 as above.With reference now to
Fig. 5-10 describes manufacture method.Most typically, such method is round manufacturing (step in the working lining 10,60 of device 100
S20) basic step of the array 30 in pearl trapping chamber 20.With previously described device 100 as one man, this manufacture ensures in chamber 20
Each there is the taper limited by one or more sidewall 21-24, described sidewall is all hydrophilic.Chamber 20 is all as blind hole
The thickness of working lining 10,60 extends.
Depending on selected material 10,60, distinct methods is possible.Chamber can be located in substrate layer 10 or cap rock or
In film 60.As previous it is further described that manufacture method can include anisotropic etching process, thermal imprint process or any other
Suitably technique is to obtain chamber 20.Up to now, anisotropic etching process in terms of the quality obtained for chamber
Promising.Base etch process when the anisotropic etching process of limit is better than, reason is that the former is not easily susceptible to etch-rate
Change and allowable depth keeps nearly constant (the last degree of depth is by the size limitation of chamber opening 28) in the case of overetched.
As it was earlier mentioned, anisotropic etching process preferably on<100>wafer perform,<100>wafer have<
110 > par on direction；Therefore top surface has the normal on<100>direction.The exposure of wafer is accordingly parallel to
(100) plane, is i.e. orthogonal to (100) direction (being diamond structures for Si) on the basis of reciprocal lattice vector.Except chamber
Outside manufacture, anisotropic etching process can be used for manufacturing microfluidic structures (such as, Fig. 8 and 9).If having inclined side
Wall is not harmful for microfluidic structures (such as passage), then the wet etching of the Si wafer with<100>crystal orientation is better than dry corrosion
Lithography, to be wet etching compatible with batch processing and therefore can the most faster for reason, and this depends on processed wafer
Quantity.It should be noted that, wet etching is generally slow than dry ecthing, producing each wafer, dry ecthing is faster.Therefore total output takes
Certainly in the quantity of the wafer processed together.
If desired, (one or more) array 30 can cleaned (such as, use ethanol, water etc.) and/or use plasma
Body (such as, air, oxygen or helium) processes.In all cases it is possible to only by by the droplet spreads S32 of pearl solution 55 to example
As on the top of (one or more) array 30 by pearl 50,51,52 deposition S30 in chamber 20.For example, it is possible to by about 2 μ L
Deposit pearl solution (being provided typically as 1% solid suspension) be applied to (one or more) array 30.
After deposition S30 pearl (and before sealing S40 chip), it is proposed that:
-it is dried S34 chamber 20 (pearl 50 be maintained in chamber or near)；And then
-such as by rinsing S36a (one or more) array 30 with rinse solution and/or being used for adhering to by applying
S36 excess pearl (that is, not being trapped in those pearls within chamber 20) is removed in bringing of S36b excess pearl；Afterwards
-if desired, after drying S38 has the array 30 in the chamber 20 of residue pearl wherein, the most always needs last dry
Dry step, such as when only applying band to remove excess pearl.
Present inventors have recognized that, before removing excess pearl, dried array 30 unexpectedly causes more stable pearl
50.Consequently found that the impact on last occupancy is significant: depend on other condition, due to drying steps S34 formerly, account for
20% to 60% can be improved by rate, the most.May infer that conical cavity is more preferable when rinsing and be dried (such as, N2 flowing)
Ground protection pearl avoids lateral, the impact of move in plane of fluid (gas or liquid).Preferably rinse solution e.g. buffers
Solution or deionized water.More generally, it can be to affect pearl or any solution of albumen being coated on.
Finally, array 30 (more generally part or all of chip 100) can seal with cap rock 60 (such as, dry film)
S40, described cap rock is preferably laminated to ensure excellent sealing.When chamber is located in layer 10, lid 60 is positioned to relative to chamber 20
Array 30 extend, be consequently formed Guan Bi microfluidic channel 12,12a, 14 and structure 16,18.
Discuss four different manufacture examples the most in detail.First manufactures example is illustrated in detail in Figure 5.Fig. 5's is detailed
Thin step is additionally captured in the flow chart of Figure 10.Fig. 6 only illustrates the selected step of this first manufacture method, and Fig. 7-9
The selected step of other possible manufacture method is shown.In each of Fig. 6-9, upper row's figure is the miniflow in the different fabrication stages
The sectional view of body chip, and lower row shows corresponding top view.
With reference first to Fig. 5, Fig. 6 and Figure 10, first manufactures example uses the etching of anisotropy chamber, is then used by dry film against corrosion
Agent passage manufactures.In detail:
-step S10: providing micro-fluid chip main body, it includes layer (or substrate) 10.
-block S20: chamber and the manufacture of passage:
Zero S21, Fig. 5 A: layer 10 is oxidized, such as, silicon is oxidized to obtain SiO2 layer 10o by thermal oxide.The electricity obtained
Insulating barrier 10o typically covers whole substrate 10.Replace oxide, it is also possible to attempt to obtain nitride, such as Si3N4.
Zero in step S22, Fig. 5 B: such as use dry and wet etching or wet etching patterned oxide.For this purpose, photic
Resist typically serves as mask (Fig. 5 B is schematically depicted in etching oxide and peels off the chip after photoresist)；
Zero then, anisotropic etching substrate layer 10, step S24, Fig. 5 C, thus obtains chamber 20 and other deposition region
70.Preferably, using wet etchant, it is typically TMAH or KOH.Typically before Si etches, carry out short oxide etching
(such as BHF) is so that the native oxide removed on Si surface；
Zero then: peels off oxide, step S25, Fig. 5 D.But this is optional, reason is that when needing, oxide is permissible
Remain on the surface.Typically, buffer oxide etch (such as BHF) is for the purpose；And preferably by deposition, expose and
Then develop dry film photoresist (negative photoresist) or epoxy-based negative photoresist (such as SU-8) patterning passage
Sidewall 52, step S26, Fig. 5 D；
-block S30: pearl deposition (also referred to as reagent integration):
Zero step S32, on chamber array 20, (different types of pearl can be deposited on phase to Fig. 5 E: the droplet deposition of pearl solution
On the array answered, as shown in Figures 12-13)；
Zero step S34, Fig. 5 F: dried array 30 and pearl are (preferably by natural evaporation, by the air-flow of N2 or by inciting somebody to action
Chip is placed in controlled environment or in warm plate, etc.)；
Zero step S36: remove excess pearl, preferably pass through: (i) rinses them, such as, use deionized water or buffer molten
The stream of liquid, step S36a, Fig. 5 G；And/or (ii) applies the band of such as adhesive tape or PDMS, excess pearl will adhere to described band, step
Rapid S36b, Fig. 5 H.If desired, band is repeatedly applied；And
-S40, Fig. 5 I: last, chip 100 (particularly (one or more) array 30 in chamber) is with being preferably laminated S42
Cap rock 60 (such as, dry film photoresist) seal.For this purpose, cap rock 60 is typically by appropriate heating to such as 45-50 DEG C.
Epiphragma can have the opening at loading pad to move liquid liquid (seeing in step s 40, Fig. 6).Can be by cutting or punching
Pressure patterns such opening.
The example above of manufacture method allows flexible design, and reason is that the manufacture of chamber and passage is separated.Can also recognize
Arriving, such method allows circular passageway configuration.But the defect of such method is that it needs twice mask and is rushed
The pearl washed may stay channel interior.
With reference now to Fig. 7, the second manufacture example described, wherein integument be incorporated into cap rock 60 (such as, dry film photoresist) and
It it not layer 10.Briefly:
-step S20a: use mould 65 to pattern chamber (such as, passing through hot padding) in layer 60；
-step S20b: separate the layer 60 of patterning；
-step S30: pearl is deposited on patterning again by moving liquid pearl solution droplets, dry, flushing and after drying
In the chamber of layer 60；
-step S40: last, by cap rock 60 is placed on chip, preferably by cap rock 60 is laminated on it close
Envelope MF chip.
In the method, the integration of pearl is unrelated with substrate and passage, the bigger spirit of the design of this permission substrate 10 and passage
Activity.MF chip can have and be etched (anisotropic etching, or deep reaction etching) or deposition (dry film photoresist, SU-8
Deng) or by hot padding or the microfluidic structures of moulded pattern.Epiphragma can be laminated to be with or without surface in advance and modify
Or on any compliant substrate processed.But, the method may need transparent substrates for more effective fluoroscopic examination.And
And, compared to the pearl being incorporated in substrate, pearl may be easier to depart from from film.
Then, with reference to Fig. 8, easily describing the 3rd manufacture example, it uses two step photoetching and anisotropy Si to etch:
-step S20c: carry out etched channels by the etching of anisotropy Si.Then thermal oxide (not shown) is carried out；Then
-step S20d: carry out etched cavity by the etching of anisotropy Si after the patterning of oxide；
-step S30: integrate pearl；And
-step S40: encapsulating chip.
Such manufacture method allows flexible design, and reason is that trap and passage manufacture are separated.But, it needs more
Manufacturing step (needing twice mask), and between two etching steps additional heat oxidation.It addition, in the method, quilt
The pearl rinsed may stay channel interior.
Finally, and with reference to Fig. 9, describing and finally manufacture example, it depends on single step photolithography and the etching of anisotropy Si.?
Step S20e: use anisotropic etching etched channels in a single photolithographic step and chamber in the identical time.Then, in step
S30 integrates pearl, and encapsulating chip, step S40.Utilize such method, an etching step manufactures passage and trap
Both, its cost benefit is higher.And, passage can be limited by the width of regulation passage in the etch process from limit deep
Degree, reason is that broader opening causes deeper passage, and vice versa.Each chamber can be connected to each other also via microfluidic channel
And it being connected to microfluidic networks, the width of described microfluidic channel is less than the width in chamber, as shown in S20e (Fig. 9).But, this
The manufacture method of sample means tightened up design rule, and the small size of the microfluidic channel owing to being interconnected in chamber causes higher
Flowed friction.Also there is the risk being internally formed bubble in chamber.
At present, the example of the application of embodiments of the invention is discussed.It is, in open channel system (in order to simply
For the sake of, the unused epiphragma of chip seals) middle simple ligand-receptor mensuration of demonstrating:
Use from Bangs Laboratories Inc.TM" Superavidin " coating pearl of polystyrene
(10 μ m diameter): coat 10 μm polystyrene with avidin (the known a kind of albumen being attached to the most by force biotin)
Pearl.Commercial stock solution comprises the pearl as 1% solid；
With PBS+0.5% polysorbas20, stock solution is diluted to 1/5 (can also use bigger dilution)；
Use the array in the chamber obtained according to Fig. 5 A-5C (the step S10 to S24 in Figure 10): by molten for the pearl of about 2 μ L
Liquid is dispersed on array and is dried (within 1 to 2 minutes, being enough to be dried)；
Array and pearl (about 30mL, lasting 10s) is rinsed flowing down of PBS+0.5% polysorbas20；
Then under DI current, rinse array and pearl (about 30mL, lasting 10s)；
Then dried array is flowed down at nitrogen；
A covering array and pearl with the 1%BSA+0.5% polysorbas20 in PBS continue 15 minutes；
Rinse array and pearl with PBS+0.5% polysorbas20, water and flow down at nitrogen dry.
Finally, can seal and storage chip.
Measuring for typical case, user carries out three below step, and wherein analyte is represented by fluorescence-labeled bio element molecule.
Array and pearl are exposed to 50 μ g/mL biotin-590-AttoSolution continue
15 minutes, it is protected from light simultaneously.During this step, biotin-590-Atto is attached to the avidin on pearl；
Array and pearl is rinsed with PBS+0.5% polysorbas20 and water.These rinsing steps are optional, but can be answered
With to increase the sensitivity measured.
The avidin using fluorescence microscope to monitor that biotin-Atto-590 (part) is attached on pearl (is subject to
Body).Single pearl chamber allows simple signal-obtaining and explanation not to have any optical artifacts.
Method and apparatus specifically described herein can use in the manufacture of micro-fluid chip.Consequent chip is permissible
By manufacturer in original form (such as, as structurized bilayer device) or with packaged form distribution.In the case of the latter
Chip may be mounted in single-chip package.Under any circumstance then chip can be integrated with other element, as in the middle of (a)
Product or the part of (b) final products.
In a word, embodiments of the invention provide various advantages.Such as:
-embodiments of the invention allow by especially and the most stable in the way of keep pearl, if especially existing
It is dried pearl and chamber array before removing excess pearl.A small amount of pearl is only lost during being subsequently exposed to liquid and being dried.To with
After the research of the whole world for many years of the microfluidic device measured and patterning receptor, finally find for pearl (or receptor) is integrated
Very effective ways to microfluidic device；
-current design and corresponding manufacture method also allow for placing pearl before covering trap；
The array in-chamber can determine size relative to pearl thus each chamber trapping list pearl.Single pearl chamber allows simpler letter
Number read and explain, need monolayer to accommodate pearl, and cause less (even without) optical artifacts；And
-manufacture method presented herein allows high production rate manufacture.Various pearl integrated strategies are suggested, and it allows miniflow
Body device and the flexible design of flow path.The concept that embodiments of the invention are based on by accident with some microfluidic features
(mirror, plastic chip manufacture etc.) is compatible.
Although describe the present invention with reference to the embodiment of limited quantity, modification and accompanying drawing, but those skilled in the art
It will be appreciated that various change can be carried out and equivalent can be replaced without deviating from the scope of the present invention.Especially, specifying
In embodiment, modification narration or the feature (device or method) that shows in the drawings can with in another embodiment, modification or figure
Another feature combination or replacement another feature, without deviating from the scope of the present invention.Therefore it is anticipated that about above example or
The various combinations of the feature that any one in modification is described, it is still in the range of attached claims.Furthermore it is possible to
Carry out many small amendments so that particular case or material are adapted to the teachings of the present invention scope without deviating from it.So,
It is contemplated that be unlimited to disclosed specific embodiment, but the present invention will include falling the scope in attached claims
Interior all embodiments.Furthermore it is possible to expect to be different from other modification of many being explicitly described above.Such as, it is different from herein
In other material of being expressly recited may be used for each of layer 10,60.It addition, passage, loading pad, passage, chamber etc. are permissible
With various sizes.
Reference numerals list
10 basic units
10o oxide skin(coating)
11 load pad
12 microfluidic channel (microchannel)
12a channel portion
14 multiplexing microchannels
16 capillary pumps
18 passages
20 pearl trapping chambeies
The sidewall in 21-24 chamber
The wall of 24a-d structuring (multiaspect)
28 chamber openings/tapered base
29 chamber basal surfaces
The array in 30 pearl trapping chambeies
32 thorax groups (row/column)
50 pearls
51 detection of analytes pearls
52 control pearl
55 pearl solution (drop)
60 cap rocks (dry film), lid
62 conduit walls
65 for imprinting the mould in chamber in cap rock 60
70 are used for detecting the chamber that antibody (dAb) deposits
The detection antibody (dAb) of 72 depositions
100 micro-fluid chips
Claims (15)
1. a micro-fluid chip (100), including layer (10,60), the array (30) in pearl trapping chamber (20) be located at described layer (10,
60), in, each in wherein said chamber (20) has the taper limited by one or more sidewalls (21-24), described sidewall
It is all hydrophilic, and each in wherein said chamber (20) extends as blind hole in the thickness of described layer (10,60).
Micro-fluid chip the most according to claim 1 (100), at least some in wherein said chamber (20) has by sidewall
(21-24) pyramid shape formed, described sidewall is all hydrophilic, and wherein it is preferred to, this pyramid shape is substantially by extremely
Few four sidewalls (21-24) limit.
Micro-fluid chip the most according to claim 1 and 2 (100), wherein said layer (10) include one of such as silicon or
Multiple semiconductor elements, and the pyramid shape of described chamber (20) have with the manufacture in the described chamber in described layer (10) each to
The geometry that anisotropic etch process is consistent.
4. according to the micro-fluid chip (100) described in any one in claims 1 to 3, in wherein said chamber (20) at least
One, preferred major part be all filled with pearl (50,51,52), preferably microballon, described pearl averagely has between 1 to 40 μm, preferably 2
Diameter between 20 μm and between more preferably 2 to 10 μm.
Micro-fluid chip the most according to claim 4 (100), the major part in the chamber (20) of wherein said array (30) is all
Being filled with only one pearl (50,51,52), described pearl preferably averagely has between 1 to 40 μm, between preferably 2 to 20 μm
And the microballon of the diameter between more preferably 2 to 10 μm.
6. according to the micro-fluid chip (100) described in claim 4 or 5, the average chi of the opening (28) of wherein said chamber (20)
The ratio of the average diameter of the pearl (50,51,52) in very little and described chamber (20) is between 1.0 to 2.4, and preferably 1.4
Between 2.0.
7. according to the micro-fluid chip (100) described in any one in claim 4 to 6, wherein said chamber (20) average deep
Degree is at least 0.5 with the ratio of the average diameter of the pearl (50) in described chamber (20), preferably at least 1.0, and more preferably at least
1.3。
8. according to the micro-fluid chip (100) described in any one in claim 1 to 7, in wherein said chamber (20) at least
Two or more subgroups (32) of one are connected by least one microchannel (14), and described subgroup is preferably described chamber (20)
The row or column of array (30), and wherein, it is highly preferred that one or more at channel portion (12a) in described subgroup
In be defined, diapire or the roof of described channel portion are formed by the surface of described layer (10).
9., according to the micro-fluid chip (100) described in any one in claim 1 to 8, wherein said array (30) is by relatively
The cap rock (60) extended in the array (30) of described chamber (20) seals.
10. according to the micro-fluid chip (100) described in any one in claim 1 to 10, including one or more pearl trappings
Some arrays (30) in chamber (20), wherein said array is preferably interposed between different pairs of channel portion (14).
11. micro-fluid chips according to claim 10 (100), including being correspondingly situated in described some arrays (30)
The different types of pearl of at least two (51,52) in one or more chambeies of at least two array, wherein said at least two is not
Pearl preferably difference in terms of size, coating, material and/or color with type.
12. 1 kinds of manufacture methods according to the micro-fluid chip (100) described in any one in claim 1 to 11, including:
(S10) is provided to have the micro-fluid chip main body of layer (10,60)；And
The array (30) of (S20) pearl trapping chamber (20) is manufactured, each tool of wherein said chamber (20) in described layer (10,60)
Having the taper limited by one or more sidewalls (21-24), described sidewall is all hydrophilic, and wherein said chamber (20) every
One extends in the thickness of described layer (10,60) as blind hole.
13. methods according to claim 12, wherein manufacture (S20) described array (30) and include being preferably used from limit
Anisotropic etching process is etched anisotropically through (S24) described layer (10) to obtain described chamber (20).
14., according to the method described in claim 12 or 13, also include:
By spreading the drop described chamber (20) by pearl (50) deposition (S30) to described array (30) of (S32) pearl solution (55)
In；And
(S40) described array (30) is sealed with the cap rock (60) being positioned to extend relative to the array of described chamber (20) (30), and
And wherein sealing (S40) described array (30) preferably includes lamination (S42) described cap rock (60).
15. methods according to claim 14, wherein after deposition (S30) pearl (50) and before sealing (S40),
Described method the most also includes:
It is dried the array (30) that (S34) has the described chamber (20) of pearl (50) wherein；
Preferably by rinsing (S36a) described array (30) with rinse solution and/or being used for adhering to (S36b) excess by applying
The excessive pearl that (S36) is not trapped in the inside in described chamber (20) removed by the band of pearl；And
If desired, after drying (S38) has the array (30) in described chamber (20) of pearl wherein.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB1405402.7 | 2014-03-26 | ||
| GB1405402.7A GB2524541A (en) | 2014-03-26 | 2014-03-26 | Microfluidic chip with conic bead trapping cavities and fabrication thereof |
| PCT/IB2015/051579 WO2015145280A1 (en) | 2014-03-26 | 2015-03-04 | Microfluidic chip with conic bead trapping cavities and fabrication thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106104271A true CN106104271A (en) | 2016-11-09 |
| CN106104271B CN106104271B (en) | 2018-04-20 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201580012689.2A Expired - Fee Related CN106104271B (en) | 2014-03-26 | 2015-03-04 | Micro-fluid chip and its manufacture with conical bead trapping chamber |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JP2017512987A (en) |
| CN (1) | CN106104271B (en) |
| DE (1) | DE112015000920B4 (en) |
| GB (2) | GB2524541A (en) |
| WO (1) | WO2015145280A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109701671A (en) * | 2018-12-27 | 2019-05-03 | 复旦大学 | A kind of microlayer model array chip and production and preparation method thereof |
| WO2020062149A1 (en) * | 2018-09-29 | 2020-04-02 | 天津大学 | Flexible encapsulation structure and manufacturing method therefor |
| CN112723303A (en) * | 2020-12-14 | 2021-04-30 | 苏州拉索生物芯片科技有限公司 | Microbead chip and spin coating preparation method thereof |
| CN114308163B (en) * | 2021-12-31 | 2024-01-09 | 北京京东方技术开发有限公司 | Microfluidic chip detection cartridge |
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| GB201610426D0 (en) | 2016-06-15 | 2016-07-27 | Q-Linea Ab | Sample holder for image based analysis of samples |
| EP3601595A4 (en) * | 2017-03-21 | 2020-12-16 | Muwells, Inc. | Sealed microwell assay |
| CN107583692B (en) * | 2017-05-23 | 2022-11-11 | 深圳市博瑞生物科技有限公司 | Liquid drop micro-fluidic chip and preparation method thereof |
| CN111295357A (en) | 2017-10-31 | 2020-06-16 | 阿斯特瑞格诊断公司 | Microfluidic device for cell characterization |
| DE102018203047A1 (en) * | 2018-03-01 | 2019-09-05 | Robert Bosch Gmbh | Microfluidic device |
| US11173486B2 (en) | 2019-02-13 | 2021-11-16 | International Business Machines Corporation | Fluidic cavities for on-chip layering and sealing of separation arrays |
| WO2023086216A2 (en) * | 2021-10-28 | 2023-05-19 | Worcester Polytechnic Institute | Gravity independent liquid cooling for electronics |
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Also Published As
| Publication number | Publication date |
|---|---|
| GB2548941A (en) | 2017-10-04 |
| GB2548941B (en) | 2020-08-19 |
| DE112015000920B4 (en) | 2021-09-30 |
| GB201405402D0 (en) | 2014-05-07 |
| GB2524541A (en) | 2015-09-30 |
| GB201617833D0 (en) | 2016-12-07 |
| WO2015145280A1 (en) | 2015-10-01 |
| CN106104271B (en) | 2018-04-20 |
| DE112015000920T5 (en) | 2016-11-24 |
| JP2017512987A (en) | 2017-05-25 |
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