CN101474541A - Integrated chip and device thereof, and method for preparing micrometre level dispersoid - Google Patents

Abstract

The invention discloses an integrated chip, a device adopting the integrated chip and a method for preparing micron-sized dispersoid; the substrate of the integrated chip is the substrate with hydrophilic surface; two layers of adjacent structures are a group of structures, and the integrated chip comprises at least one group of structures; in one group of structures, the first structure is provided with M levels of gradients, and the second structure is provided with N levels of gradients; wherein, M and N are natural numbers, and M is less than N; among the gradients, the primary gradient is provided with H1 channel 1, H2 branches are led out from each channel 1 in the secondary gradient, and H1*H2...*HN channel N are formed; one side which is far away from each channel M is provided with a connecting channel respectively; the integrated chip also comprises at least a collecting groove; the integrated chip also comprises at least one collecting groove which is connected with the output end of each collecting channel for collecting and exporting products. The invention has very high productivity and absolute advantage in the aspect of the large scale production of contrast agent.

Description

Integrated chip and device thereof and the method for preparing micrometre level dispersoid

[technical field]

The present invention relates to contrast agent application contrast imaging technique, in particular, a kind of large scale integrated chip (LSI chip) that adopts microflow control technique to make micrometre level dispersoid; A kind of device that adopts this chip can be used for preparing acoustic contrast agent; And adopting this device to prepare the method for micrometre level dispersoid, micrometre level dispersoid can be used as acoustic contrast agent.

[background technology]

In clinical diagnosis, the 26S Proteasome Structure and Function information that do not have wound, in real time, dynamically detects related organization has become and has become more and more important.The Ultrasonic Diagnosis technology not only has above-mentioned characteristics, and, compare with X ray, nmr imaging technique, also have safety, adapt to wide, can check repeatedly, strong to the distinguishing ability of soft tissue, flexibility reaches advantages such as cheap by force, is most popular in the world image technology.

Contrast agent application contrast video picture is the core content of image technology development.Compare with various image technologies, ultrasonic is the most extensive but only image technology of failing conventional contrast agent application of range of application, and the core of contrast echo technology is an acoustic contrast agent.

Nineteen sixty-eight, first observed such as Gramiak strengthen, have opened the prelude of cardiac acoustic contrast to through conduit injection gassiness salt solution the right heart being developed.In this stage, people mainly utilize methods such as shake by hand physiological saline or CO2 blowing agent to make acoustic contrast agents.Its microvesicle diameter is big, homogeneity is not good, is difficult to by pulmonary circulation.In addition because uncanned protection, in the microvesicle gas rapidly in blood disperse make that the microvesicle time-to-live is of short duration, have the scholar with the contrast preparation in this period be called the 0th generation acoustic contrast agent.

The employing sound methods of shaking such as Feinstein in 1984 prepare stable microbubble and just really enter the left cardiography epoch.What begin to occur most is the microvesicle of parcel air, and the sheathing material of parcel air is of a great variety, and roughly can be divided into: protein coat, liposome shell, surfactant shell and high polymer shell etc. are called first generation acoustic contrast agent.Because solubility and the spread of air in blood be bigger, the air in the microvesicle is gone into blood by the shell disperse easily, makes contrast agent microbubble diminish or disappears.In addition, because the scattered signal of microvesicle is directly proportional with 6 powers of diameter, as long as there is a spot of air to overflow, the diameter of microvesicle slightly diminishes, and the echo signal of microvesicle is obviously decayed.Representative product has the Levovist of the Albunex of the U.S., German Schering company, the stability and the time-to-live of the acoustic contrast agent of the first generation fall short of, limit its application greatly, withdrawed from contrast preparation market and research field after 2000 gradually.

Since nineteen ninety-five, because the introducing of the fluorine carbon fluorine sulphur class gas of HMW, low blood dissolves degree and dispersivity, time-to-live of acoustic contrast agent and stability have been had significantly improve, be called second generation acoustic contrast agent.The Optison of the representative acoustic contrast agent U.S.,

, EchoGen and AI-700 etc., the SonoVue (BR1) of Italian Bracco company and BR14 etc.

At present, existing abroad multiple new generation product listing, and the domestic product that not have autonomous listing, the production of Italian Bracco company be uniquely to be ratified commercial acoustic contrast agent by China Ministry of Public Health, price is very expensive.

Generally speaking, conventional acoustic contrast agent is a kind of suspension that contains the high concentration microvesicle, microvesicle is a shell with albumin, lipid, surfactant or high polymer, the about 2-4 micron of average diameter, less than red blood cell, system is felt concerned about on the pulmonary circulation arrival left side of passing through that can be stable after the PeV injection, and then arrives at each organ of whole body.The acoustic impedance difference of acoustic contrast agent microvesicle and blood and tissue is very big, and the effect between the ultrasonic wave is very complicated, can change backscattering coefficient, attenuation coefficient, the velocity of sound and other acoustic characteristic at position, place, can strengthen the ultrasonic echo ability of local organization behind the injection blood vessel, thereby obviously improve the development effect of ultrasonic image to tissue blood flow's perfusion, significantly improve the sensitiveness and the specificity of Ultrasonic Diagnosis, break through the bottleneck of conventional ultrasonic development, further expanded the Ultrasonic Diagnosis field.

And, acoustic contrast agent not only can be used for conventional radiography to be strengthened, also can be used for targeting diagnosis and PCI, ligands specific is attached to the contrast agent microbubble surface, make it arrive interested tissue or organ, optionally with corresponding receptors bind, thereby reach the purpose that specificity strengthens target area ultrasonic signal or local targeted therapy.And the research of PCI constantly enlarges the range of application of acoustic contrast agent, and using value constantly promotes.

For example, utilize contrast agent microbubble to reduce the ultrasonic cavitation threshold value, thereby promote the result of treatment of ultrasound thrombolysis or raising high-intensity focused ultrasound; Or utilize ultrasonic wave to destroy the contrast agent microbubble of carrying gene or medicine, and make the release of gene or drug targeting and promote it to penetrate, shift or the drug targeting treatment with mediated gene; Or utilize radiography guiding radio frequency or micro-wave therapeutic and estimate result of treatment etc. immediately.

The research of targeted ultrasound contrast agent is just becoming the focus that ultrasonic image is learned the field.No matter from the selection of active group, the invention of method of modifying, the detection of targeting, or medicine has all been obtained very gratifying achievement for drug efficacy study and clinical practice in the body.Constantly perfect along with the fast development of macromolecular material and microvesicle preparation technology, microcapsular ultrasound contrast agent will develop towards personalized more direction, following microcapsular ultrasound contrast agent can not only be applied to the special ultrasonic imaging under different physiology and the pathological state, can also open up the new world of a slice in the treatment field as the carrier that carries medicine or therapeutic gene.For this reason, external medical institutions prediction acoustic contrast agent will become the common agents in Ultrasonic Diagnosis or the treatment soon, have huge social benefit and economic worth.

At present, the focus of ultrasonic area research is concentrated both ways: the one, and the imaging technique of various enhancing ultrasonic contrasts, as: trigger imaging technique, Second Harmonic Imaging technology, subharmonic imaging technique, first-harmonic pulse counteracting imaging technique, energy pulse inverse imaging technology, low mechanical index Real Time Imaging Technology etc.; Then be the new more reliable and more stable acoustic contrast agent of research on the other hand.

Desirable acoustic contrast agent should possess following characteristic: (1) safety non-toxic, and viscosity is low, and the inanimate object activity does not have physiological influence to the circulatory system, not obvious dilution red blood cell concentration; (2) the microvesicle size meets the requirements, and has certain pressure in bubbles, makes the perfusion back keep sufficiently long time and enough strong echo strength; (3) reflectivity is good, and the decay artifact is little, and echo-signal and Doppler signal strengthen obviously, harmonic resonance can take place increase the back scattering area; (4) can intravenously administrable, the body internal stability is good, and is easy to use, and the instrument condition of radiography video picture is not had very special requirement, is convenient to popularize; (5) have stable concentration and formulation, be easy to sterilization, sterilization, preserve, transport, can produce in batches.

At present the most frequently used method for preparing the acoustic contrast agent microvesicle is shake method and a mechanical oscillation method of sound.

The sound method of shaking is the positive and negative acoustic pressure of the high frequency conversion that produces when utilizing supersonic oscillations, negative acoustic pressure wherein makes and is present in the gas that contrast preparation prepares in the liquid and expand forms micro-bubble, at this moment, lipid in the preparation liquid or albumin, surfactant, polymer etc. are taken advantage of the occasion to wrap up micro-bubble and are formed stable contrast agent microbubble.Albumin class in the acoustic contrast agent, lipid, polymer class, surfactant-based etc., its preparation process all are the employing sound methods of shaking.

But there is following shortcoming in the sound method of shaking: the technological parameter of (1) sonde-type sound Vibration Meter, comprise power, probe in the position of liquid level, the degree of depth etc., wayward, the technology reappearance is under some influence; (2) because probe will be placed on preparation liquid inside, therefore in sound shakes process, be difficult to accomplish sterile working, and have the possibility of metallic pollution, increased certain difficulty for quality control, the preparation technology of contrast preparation; (3) sound shakes and produces more heat in the process, and to the activity of lipid, especially when the contrast preparation of some part, medicine or gene is carried in preparation, the activity of part, medicine or the gene that sound is simultaneously shaken produces very big influence.

The mechanical oscillation method is when utilizing the high frequency mechanical oscillation, in the preparation liquid each point stressed the time differently mutually produce different positive/negative pressures, negative pressure wherein can make the gas that is present in the preparation liquid form micro-bubble.Frequency is high more, and the conversion of positive/negative pressure is fast more, and the time of negative pressure is short more, and gas expands more little, and the bubble of formation is just more little; Amplitude is more little, and the negative pressure of generation is more little, under the identical time situation, more little by the gas expansion that negative pressure produces, the bubble of formation is more little, because the preparation acoustic contrast agent need form less microvesicle, therefore, the mechanical oscillation device of preparation acoustic contrast agent needs higher frequency, lower amplitude.

But there is following shortcoming in the mechanical oscillation method: (1) contrast agent microbubble particle size can not accurately be controlled; (2) the microvesicle particle diameter that forms distributes and compares broad, the acoustic characteristic instability, all contain some in the microvesicle greater than 10 microns microvesicle, these big microvesicles may cause the obstruction of local vascular or break, and make the use of acoustic contrast agent hide certain risk in the hiding; (3) outer casing thickness of formation microvesicle is inhomogeneous; (4) the mechanical oscillation method produces enough power and gas is incorporated into from surrounding environment forms bubble the liquid solution, hunting speed determines to form the quantity and the size of microvesicle to a great extent, contrast preparation SonoVue as domestic clinical approval use, use hand mode to prepare, repeatability is good inadequately.

Therefore, there is defective in prior art, needs to improve.

[summary of the invention]

Technical problem to be solved by this invention provides a kind of extensive micro-fluidic flow focusing apparatus of practicality, and uniform particle diameter, can control, acoustic contrast agent preparation method that monodispersed acoustic properties is stable, controlled and be the acoustic contrast agent of monodispersity with the preparation particle size.On this basis, provide a kind of integrated chip, adopt the device of this integrated chip and adopt this device to prepare the method for micrometre level dispersoid.

Technical scheme of the present invention is as follows:

A kind of technical scheme of integrated chip is, is one group of structure with two-layer adjacent structure, and described integrated chip comprises at least one group of structure; In one group of structure, the ground floor structure is provided with M level gradient, and second layer structure is provided with N level gradient; Wherein, M, N are natural numbers, and M is less than N; In the gradients at different levels, first order gradient is provided with H1 passage 1, and second level gradient is drawn H2 branch from each passage 1, form H1 * H2 passage 2, by that analogy, N level gradient is drawn HN branch from each passage (N-1), forms H1 * H2...... * HN passage N;

In one group of structure,, all formed H (M+1) * H (M+2) from what a passage M drew in N level gradient ... the special modality N of * H (N-1) * HN quantity corresponding to second layer structure for arbitrary special modality M of ground floor structure;

Wherein, the outlet of each special modality N is equally distributed on a circumference that pre-sets radius circular; For this special modality M, its outbound course is perpendicular to this circle, and its outlet is passed through this circular center of circle with this circular vertical line, and the outlet of this special modality M and this distance of center circle are from a length-specific; The outbound course of each special modality N is identical acute angle with the angle of described vertical line; Along the outbound course of this special modality M,, a collection channel is set in the side of this circle away from this special modality M; Described integrated chip also comprises at least one feeder that is connected with the output of each collection channel, is used for collecting and deriving product.

Another technical scheme of integrated chip is M=N-1.

Another technical scheme of integrated chip is that the outbound course of each special modality N is by this circle center of circle.

Another technical scheme of integrated chip is that the cross section of passage M and passage N is a rectangle; Described rectangle height is that 20 μ m to 30 μ m, width are respectively 30 μ m to 50 μ m, 50 μ m to 100 μ m, perhaps, is respectively 50 μ m to 100 μ m, 30 μ m to 50 μ m.

Another technical scheme of integrated chip is, the structure of described collection channel comprise at least following structure one of them: export-oriented cone, spheroid, hemisphere or the cone table that amplifies, the caliber of its input is 7 μ m to 25 μ m, highly is 20 μ m to 30 μ m; Perhaps, cylinder or cuboid, its sectional area along the special modality M outbound course of correspondence is slightly less than this special modality M sectional area in the direction, highly is 20 μ m to 30 μ m.

Another technical scheme of integrated chip is that its matrix is surface hydrophilicity matrix.

Another technical scheme of integrated chip is that its matrix is silicon, glass, dimethyl silicone polymer, polymethyl methacrylate or poly-carbon ester.

Another technical scheme of integrated chip is that the combination of ground floor structure and second layer structure is the combination of pathway structure and fluid passage structure.

A kind of technical scheme of device is that it comprises: integrated chip array, storage unit that the first material input block, the second material input block, at least one integrated chip form; Wherein, integrated chip is above-mentioned arbitrary technical scheme about integrated chip, pairing integrated chip;

The feeder of each integrated chip is connected with described storage unit respectively, is used for collecting and deriving product; The described first material input block is connected with each passage 1 of one deck structure of each integrated chip respectively, is used to import first material; The described second material input block is connected from each passage 1 of another layer structure of each integrated chip respectively, is used to import second material.

Another technical scheme of device is that the combination of described first material input block and the described second material input block is the combination of gas input block and liquid input block; Perhaps, described first material input block and the described second material input block are the liquid input block.

Another technical scheme of device is, described gas input block comprises pressure gas tank, pressure-reducing valve, first transfer tube, micro-flowmeter, pressure regulator valve and second transfer tube that is linked in sequence, and described second transfer tube is connected with each passage 1 of one deck structure of each integrated chip respectively; Described liquid input block comprises reservoir, the 3rd transfer tube, digital control type syringe pump and the 4th transfer tube that is linked in sequence, and described the 4th transfer tube is connected with each passage 1 of another layer structure of each integrated chip respectively.

A kind of technical scheme for preparing the method for micrometre level dispersoid is, be applied to one and comprise in the device of integrated chip array that described integrated chip array is made up of integrated chip, wherein, integrated chip is above-mentioned arbitrary technical scheme about integrated chip, pairing integrated chip; Device is above-mentioned arbitrary technical scheme about device, pairing device;

Said method comprising the steps of: A1, pre-set condition by first, to each passage 1 input first material of one deck structure of each integrated chip; Pre-set condition by second, to each passage 1 input second material of another layer structure of each integrated chip; The initial conditions of A2, adjusting first material and/or second material, the material of each passage M output that makes each ground floor structure is with coaxial liquid form,, formed respectively and wait to collect product stream by axially wrapping up by the material of each special modality N of correspondence, it axially overlaps with described vertical line; A3, collect and derive product from the output of each collection channel.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is that the combination of described first material and described second material is the combination of gas and liquid; Described gas comprise at least nitrogen, fluorine carbon gas, fluorine sulphur class gas one of them; Described liquid comprise at least phospholipid liquid, surfactant class I liquid I, aforesaid liquid modification liquid one of them.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is, in the steps A 1, to each passage 1 input first liquid of the ground floor structure of each integrated chip, to each passage 1 input gas or second liquid of the second layer structure of each integrated chip; In the steps A 2, first liquid of each passage M output that makes each ground floor structure is with coaxial liquid form,, formed respectively and waits to collect little stream of liquid droplets by axial described first liquid of parcel by gas or second liquid.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is, in the steps A 1, to each passage 1 input gas of one deck structure of each integrated chip, pressure is lower than 10psi, to each passage 1 input liquid of another layer structure of each integrated chip, flow velocity is lower than 3 μ L/s; In the steps A 2, the initial conditions of adjustments of gas and/or liquid.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is, in the steps A 1, the pressure of input gas is lower than 5psi, and the flow velocity of input liquid is lower than 1.5 μ L/s.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is that in the steps A 1, described liquid adds ligands specific in advance; Perhaps, after the steps A 3, add ligands specific to described product.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is, in the steps A 1, to each passage 1 input gas of the ground floor structure of each integrated chip, to each passage 1 input liquid of the second layer structure of each integrated chip; In the steps A 2, the initial conditions of adjustments of gas and/or liquid, the gas of each passage M output that makes each ground floor structure be with coaxial liquid form, by axial parcel liquid, forms respectively and wait to collect microballoon stream.

The another kind of technical scheme for preparing the method for micrometre level dispersoid is, in the steps A 2, the gas of each passage M output of each ground floor structure, form the stable reverse taper that the axis is positioned at described vertical line position respectively under the swiftly flowing fluid-focus effect in both sides, the tip of each reverse taper is relative with each collection channel position respectively.

Adopt such scheme, the invention provides a kind of integrated chip, adopt the device of this integrated chip and adopt this device to prepare the method for micrometre level dispersoid.On this basis, the invention provides a kind of extensive micro-fluidic flow focusing apparatus of practicality, and uniform particle diameter, can control, acoustic contrast agent preparation method that monodispersed acoustic properties is stable, controlled and be the acoustic contrast agent of monodispersity with the preparation particle size; Easy, effective and safety that this acoustic contrast agent prepares, the microvesicle outer casing thickness of preparation is even, has very high using value.

The most significant advantage is that the acoustic contrast agent microvesicle of making has high degree of monodispersity and particle diameter controllability, satisfies the requirement of ultrasonic contrast imaging technology.The polydispersity index of microvesicle particle diameter＜2%, particle diameter become along with the increase of gas pressure greatly, reduce along with the increase of flow rate of liquid, control very flexible; Simultaneously, adopt the formed micro-nozzle array of a plurality of collection channels to improve the preparation efficiency of microvesicle greatly; Process units has reusability, has reduced production cost.

And apparatus and method of the present invention can be used for preparing polytype micrometre level dispersoid, comprise the acoustic contrast agent microvesicle, and little drop or other miniature mixtures.Conventional acoustic contrast agent can be directly with this device preparation, as lipid, albumin class, polymer class, surfactant-based etc.; By adding ligands specific, can also prepare targeted ultrasound contrast agent, some shell material, as protein molecular, inactivation under high temperature and ultrasound condition, thus the inventive method solved sound in the past shake method can not the technical problem of solution.This contrive equipment all is suitable for these two kinds of methods, particularly has more advantage when adopting a kind of selection in back, and its advantage one is to have reduced the preparation link, and the 2nd, the destruction of having reduced preparation link microvesicle.And apparatus of the present invention heat production in the contrast preparation preparation process is few, therefore especially is suitable for preparing the acoustic contrast agent of double medicine or gene targeting vector.

[description of drawings]

Fig. 1 is the schematic diagram of the acoustic contrast agent preparation facilities of the embodiment of the invention;

Fig. 2 is the bottom channel design schematic diagram of the micro-fluidic large scale integrated chip (LSI chip) of the embodiment of the invention;

Fig. 3 is the top layer channel design schematic diagram of the micro-fluidic large scale integrated chip (LSI chip) of the embodiment of the invention;

Fig. 4 is the nozzle arrangements enlarged diagram shown in Fig. 2 frame of broken lines;

Fig. 5 is that the microvesicle of the embodiment of the invention forms schematic diagram;

Fig. 6 is the schematic diagram of the micro-fluidic large scale integrated chip (LSI chip) of another embodiment of the present invention;

Fig. 7 is the enlarged drawing of structure shown in Fig. 6 frame of broken lines;

Fig. 8 is that the present invention tests a microvesicle photo that generates;

Fig. 9 is the another kind of nozzle arrangements schematic diagram of embodiment of the invention device;

Figure 10 is in the connected chamber of the embodiment of the invention, the schematic diagram on the plane, outlet place of each special modality N;

Figure 11 is the schematic diagram of fore-and-aft plane shown in Figure 10.

[specific embodiment]

Be elaborated below in conjunction with the drawings and specific embodiments.

The particle size of acoustic contrast agent microvesicle and distribution thereof are the important indicators of acoustic contrast agent, yet, present acoustic contrast agent preparation method such as the sound microvesicle particle size distribution range broad that method, mechanical oscillation method produce that shakes, and mainly form the acoustic properties instability by mode hand or vibration.Therefore, a kind of preparation method and device of extensive micro-fluidic acoustic contrast agent are proposed, make micro-fluidic large scale integrated chip (LSI chip) in conjunction with ripe at present micro-nano process technology, utilize the flow focusing principle, prepare that particle size can be controlled and the microvesicle of high degree of monodispersity, the microvesicle outer casing thickness is even, not only can obtain stable acoustic properties, also can reduce the possibility that is fused into big microvesicle between the different big or small microvesicles, have higher validity and security, operation has better repeatability and controllability.

Therefore, the present invention proposes a kind of integrated chip that adopts microflow control technique, by micro-fluidic mode, can prepare micrometre level dispersoid on a large scale, comprises acoustic contrast agent or other miniature mixtures.

The matrix of integrated chip is surface hydrophilicity matrix, and described surface hydrophilicity matrix comprises the hydrophobic base that the hydrophily processing was carried out on hydrophilic matrix and surface; Need to prove that the material of passage not necessarily will have hydrophily, still, the effect of hydrophobic material does not have the effective of hydrophilic material, for hydrophobic material, can be hydrophily with its surface treatment, can obtain to be similar to the effect of hydrophilic material yet; The surface is hydrophilic matrix, can produce microvesicle very swimmingly.

An example is that its matrix is silicon, glass, dimethyl silicone polymer, polymethyl methacrylate or poly-carbon ester.Wherein, can satisfy structural requirement on the technology, be difficult for the gas interior with passage, the material that liquid reacts all can be used as matrix, the present invention does not make any additional limits to this; For example, dimethyl silicone polymer has good insulation performance, can bear high voltage; The heat endurance height is fit to the various reaction chips of processing; Have good optical characteristics, can be applicable to multiple optical detection system; In addition, it can also form good sealing with many materials such as silicon, silicon nitride, silica, glass, and it also has high hydrophobicity and the surperficial characterization of adsorption stronger to large biological molecule, therefore has been widely used in the micro-fluidic chip research field.

With two-layer adjacent structure is one group of structure, and described integrated chip comprises at least one group of structure; Be that described integrated chip can comprise double-layer structure, perhaps comprise four-layer structure, six layers of structure, eight layers of structure even more.For example, described integrated chip only is provided with one group of structure.That is, described integrated chip is the even level structure, carries out stacked on top or inclination and piles up and form, and the concrete feature of each layer structure is as described below: the marginal layer structure of the arbitrary end of described integrated chip, and the layer structure adjacent with this marginal layer structure formed one group of structure; Described integrated chip can have one group of structure, and many group structures also can be arranged.Integrated chip also can be arranged in parallel and form, and promptly one group of structure or the sandwich construction of organizing stacked on top are arranged in parallel more.

In one group of structure, have two-layer adjacent structure, wherein, the ground floor structure is provided with M level gradient, and second layer structure is provided with N level gradient; Wherein, M, N are natural numbers, and M is less than N; In the gradients at different levels, first order gradient is provided with H1 passage 1, and second level gradient is drawn H2 branch from each passage 1, form H1 * H2 passage 2, by that analogy, N level gradient is drawn HN branch from each passage (N-1), forms H1 * H2...... * HN passage N.Preferred example is, every grade of branch will evenly distribute with respect to the branch of previous stage bifurcated, mainly is in order to guarantee the flow velocity unanimity of the same liquid that last each passage comes out, or the flow velocity unanimity of same gas; That is, N level gradient is drawn HN branch from each passage (N-1), and HN the branch that a certain passage (N-1) is drawn evenly distributes with respect to this passage.

Wherein, each passage 1 of ground floor structure is used to import first kind of material; Each passage 1 of second layer structure is used to import second kind of material.First kind of material and second kind of material can be different gas or liquid, also can also be for two kinds of different liquid, and first kind of material is gas, and second kind of material is liquid, also can be, first kind of material is liquid, and second kind of material is gas.For example, the ground floor structure is as gas passage, and second layer structure is as fluid passage; Perhaps, the ground floor structure is as fluid passage, and second layer structure is as gas passage; Perhaps, the ground floor structure is as first kind of fluid passage, and second layer structure is as second kind of fluid passage.

For example, the ground floor structure is as gas passage, and second layer structure is as fluid passage; At this moment, the H1 of a ground floor structure passage 1 connects gas input port respectively, and the H1 of a second layer structure passage 1 connects liquid inlet respectively; H1 * the H2...... of ground floor structure * HM passage M connects gas delivery port respectively, and the H1 * H2...... of second layer structure * HN passage N connects liquid outlet.Perhaps, the ground floor structure is as fluid passage, and second layer structure is as gas passage, and all the other are similar, do not do and give unnecessary details.

A preferred example is, M=N-1, and promptly M is littler by 1 than N, and the gradient number of ground floor structure lacks one deck than the gradient number of second layer structure.For example, N is 8, and M is 7.

Perhaps, M littler by 2 than N, 3,4 ... or the like, for example, N is 8, M is 4; And for example, N is 6, and M is 3; And for example, N is 9, and M is 7; And for example, N is 21, and M is 12.The present invention does not do additional limits to this, only need to realize that the difference of ground floor structure and second layer structure gradient number gets final product.

Wherein, H1, H2, H3......HM, H (M+1), H (M+2) ... H (N-2), H (N-1), HN can be all identical, also can part identical, can also be all inequality.For example, H1, H2, H3......HM, H (M+1), H (M+2) ... HN is 2, and, M=N-1, the quantity of passage N is 2 times of passage M like this; In this case, because H1 is 2, therefore two gas accesses and two liquid inlets are arranged; Perhaps, H1 is 2, and H2 to HN is 3; Perhaps, H1 is 1, and H2 is 2, H3 is 3, H4 is that 4......HN is N; Perhaps, it is all inequality, is 4 as H1, and H2 is 1, H3 is 6, H4 is that 5......HN is 2.The present invention does not do additional limits to this, only need to realize that branched structures at different levels get final product.

For the shape of passage, the cross section of each passage is preferably rectangle; Wherein, passage M cross section and passage N cross section can be shape rectangle identical or inequality, for example, the height of described rectangle is 20 μ m to 30 μ m, and the width of passage M cross section is 30 μ m to 50 μ m, and the width of passage N cross section is 50 μ m to 100 μ m; Perhaps, the width of passage M cross section is 50 μ m to 100 μ m, and the width of passage N cross section is 30 μ m to 50 μ m; Perhaps, the width of passage M cross section and passage N cross section is 30 μ m to 50 μ m.

An example is, passage is homogeneous or inhomogenous cuboid, but the cross section must be a rectangle, and for example passage is a positive tetragonous cone table, a tetragonous cone table or the bucking ladder that cross section is similar rectangle.A preferred example is, each passage is a cuboid, and the cross section of passage is a rectangle, and it highly is 20 μ m to 30 μ m, and width is 30 μ m to 100 μ m.For example, the height of this rectangle is 25 μ m, and width is 40 μ m; And for example, the height of this rectangle is 28 μ m, and width is 35 μ m; And for example, the height of this rectangle is 22 μ m, and width is 45 μ m; And for example, the height of this rectangle is 21 μ m, and width is 85 μ m; And for example, the height of this rectangle is 29 μ m, and width is 65 μ m; And for example, the height of this rectangle is 29 μ m, and width is 75 μ m.

A preferred example is that passage M cross section and passage N cross section are highly identical rectangle.

That is to say, for a certain passage, perpendicular to the cross section of its channel direction, promptly the channel cross-section of this passage is square or rectangular as described cross section, need to prove, channel cross-section can also be triangle, rhombus, pentagon or other shapes, and the present invention does not do additional limits to this; For each passage, only need the area of its channel cross-section, be at least 600 square microns and get final product, for example, the area of its channel cross-section is 800,900 or 1000 square microns.And for example, each passage is cylinder or its allothimorph; Perhaps, each passage can also be square body, round or the helical form of bending break-in, for example L-shaped, Y shape etc.; At this moment, the outbound course of passage is the not direction of bent passage of its final stage, and for example, the outbound course of L shaped passage is the direction that has one section passage of outlet after the break-in.

In one group of structure, for arbitrary special modality M of ground floor structure, all corresponding to a certain branch of second layer structure from HM the branch that a passage M is drawn; This branch forms the individual branch of H (M+1) in M+1 level gradient ... form H (M+1) * H (M+2) in N level gradient ... the special modality N of * H (N-1) * HN quantity; Promptly for arbitrary passage M of ground floor structure, in second layer structure, the second layer structure channel M corresponding with this ground floor structure channel M will draw the individual branch of H (M+1) in the next stage gradient, these branches finally form H (M+1) * H (M+2) in N level gradient ... * H (N-1) * HN passage N, M is corresponding with this ground floor structure channel.

Wherein, the outlet of each special modality N is equally distributed on a circumference that pre-sets radius circular, that is, the outlet of these passages N is arranged and formed regular polygon, and is positioned at each summit on the regular polygon; For example, these passages N's goes out interruption-forming equilateral triangle, square, regular pentagon, regular hexagon ... positive 99 limit shapes, positive 100 limit shapes, positive 101 limit shapes or the like; Special, when two passage N were only arranged, outlet was positioned at the two ends that this pre-sets a certain diameter of radius circular.Wherein, pre-seting radius can be provided with according to actual conditions, for example, according to different reactants, different reaction conditions, pre-sets the size of this circular radius.Circular radius has relation with the size of passage, and with ground floor passage input gas, it is example that second layer passage is imported liquid, and gas vent is during perpendicular to aforementioned circle, and the periphery that liquid outlet is distributed on gas vent gets final product.For example, this circular radius is 150 μ m to 1500 μ m, and example is more specifically, and this circular radius can be 250 μ m, 500 μ m, 700 μ m, 1100 μ m, 1200 μ m, 1400 μ m or the like.Preferably, this circular radius is corresponding with width of channel, and width of channel is big more, and then this circular radius is big more.

Need to prove, when yardstick hour, for example, when measuring with micron order, the outlet of passage and the input of collection channel, the notion of a face often; And in the various embodiments of the present invention, every line that relates between points is interpreted as the central point of exit face, perhaps, and the central point of the face of input; For example, the geometric center point of triangle or rectangle; Perhaps, be roughly the central point of exit face, or the central point of the face of input.Like this, outlet is arranged and is formed regular polygon, could obtain better precision.And the line between the input of the line between the outlet, outlet and collection channel could be more accurate, to obtain better effect.

An example as shown in figure 10, six special modality N are arranged, its outlet is equally distributed on a circumference that pre-sets radius circular, six go out interruption-forming one regular hexagon, are arranged in a transverse plane, and this transverse plane is arranged in a connected chamber, liquid flows to the center of circle that this pre-sets radius circular from the outlet of each special modality N respectively, that is, liquid outlet is identical to the distance in the center of circle, and is arranged as regular hexagon; And in the regular hexagon, any two relative special modality N are positioned at a fore-and-aft plane; For example any two relative special modality N form symmetry or symmetric relation roughly with this center of circle that pre-sets radius circular.

In the last example, arbitrary fore-and-aft plane as shown in figure 11, two relative special modality N for outlet and this circular vertical line of this special modality M, form the axial symmetry relation; Ground floor gas is after outlet output, continue to flow again a segment distance, i.e. one section length-specific, this length-specific mainly is that height or the width with second layer fluid passage rectangular cross section has relation, for example, this length-specific be the fluid passage cross section width or the height 25% to 75%, as 30% of the height of channel cross-section, 40%, 50%, 60%, 70% or the like, preferred example is, when having only two fluid passages and a gas passage, all outlet ports is all in a plane, and as shown in Figure 4 and Figure 5, the distance in gas passage outlet and the center of circle is exactly half of width of fluid passage cross section; As a plurality of fluid passages, gas passage outlet is during perpendicular to fluid passage outlet plane, place, and the distance in gas passage outlet and the center of circle is exactly half of height of fluid passage cross section.Gas mixes with liquid afterwards, before entering the input of described collection channel, forms and waits to collect product stream, collects by collection channel; For example, formation reverse taper or other shapes waits that collecting product flows, and collects by collection channel.

Above these two examples, description be even number special modality N, arbitrary fore-and-aft plane has and forms the axial symmetry relation or a pair of special modality N of symmetry roughly; Need to prove, if odd number special modality N, though on fore-and-aft plane, can not form a pair of special modality N,, can realize effect of the present invention equally.For example, five special modality N, its outlet is equally distributed on a circumference that pre-sets radius circular, five go out interruption-forming one regular pentagon, be arranged in a transverse plane, this transverse plane is arranged in a connected chamber, and liquid flows to the center of circle that this pre-sets radius circular from the outlet of each special modality N respectively.

The outbound course of this special modality M is perpendicular to this circle, the outlet of this special modality M and this circular vertical line are by its center of circle, the plane that is the outbound course of this passage M and circular place is perpendicular, and the projection of outlet on this plane of this passage M is positioned at this circular center of circle.The outbound course of each special modality N is identical with the angle of described vertical line, and these angles are acute angle; For example, the outbound course of each special modality N is by this circle center of circle, and each special modality N is positioned on each bisector of this circle according to its quantity, supposes to have 3 special modality N, and the angle between then adjacent two special modality N is 120 degree; Perhaps, each special modality N can form a straight cone, and this straight cone is a rotating shaft with the outbound course of this special modality M, and each special modality N is identical with the angle of this rotating shaft, and these angles are acute angle; For example, angle is 30 degree, 45 degree, 60 degree, 75 degree or 80 degree or the like.For example, because the outlet of each special modality N is equally distributed on a circumference that pre-sets radius circular, when the quantity of each special modality N is even number, be symmetrical in the outlet of two special modality N of this rotating shaft, pairing two special modality N formation one comprises the plane of this rotating shaft, and two special modality N are on its outbound course, and it is trapezoidal to form isosceles.For example, each special modality N forms a kind of shape of symmetry; For example be the hexagonal pyramid body of a half, i.e. hexagonal pyramid platform, its bottom surface is a regular hexagon, six ribs can be formed three isosceles trapezoids.

Like this, in one group of structure, by the initial conditions of adjusting one deck structure and the initial conditions of another layer structure, the material of each passage M output that makes each ground floor structure is with coaxial liquid form, by the material of each special modality N of correspondence by the axial material of a certain passage M output of parcel, promptly the flow direction with the material of a certain passage M output is an axle, by exporting the material of each corresponding special modality N, thereby forms from level to level the material coated with each special modality N output of correspondence; In other words, the material of each special modality N output covers with the concentric circles form on the material of passage M output vertically; Form respectively and wait to collect product stream, for example, wait to collect product stream and form a reverse taper; It axially overlaps with described rotating shaft.Be the material of a certain passage M output of certain ground floor structure, flow along its outbound course always, perhaps flow around its outbound course with spirality; With its outbound course serves as axially, by the material of each special modality N of its correspondence by axial parcel.

The outlet of each special modality N is communicated with setting with the outlet of this special modality M; For example, the outlet of each special modality N is communicated with setting by a connected chamber with the outlet of this special modality M; In each example of the present invention, connected chamber is not imposed any restrictions, only need to realize forming one and wait to collect product stream and get final product, this connected chamber can be set yet by feeder output.

Along the outbound course of this special modality M, in the side of this circle away from this special modality M, correspondence is provided with a collection channel; Be used for before the input of described collection channel, form and wait to collect product stream, collect product, output to feeder by collection channel at the input of collection channel; Promptly be provided with the collection channel identical with the quantity of passage M, for each special modality M, along its outbound course one collection channel be set respectively, special modality M is identical with the outbound course of its collection channel, with in a straight line.

An example is, the structure of described collection channel comprises one or more of following structure: export-oriented cone, spheroid, hemisphere or the cone table that amplifies, a cone table i.e. cone cuts most advanced and sophisticated residue stage body part, and the caliber of its input is 7 μ m to 25 μ m, highly is 20 μ m to 30 μ m; Perhaps, cylinder or cuboid, as shown in Figure 9, its area along the channel cross-section of the special modality M outbound course of correspondence is slightly less than the area of this special modality M channel cross-section in the direction, highly is 20 μ m to 30 μ m, at this moment, the length that needs control cylinder or cuboid passage, make microvesicle just in time in feeder, produce, possessing velocity gradient, thereby produce shearing force.For example, the structure of collection channel is export-oriented cone of amplifying, and perhaps, the structure of collection channel is export-oriented cone table and cylindrical combination of amplifying.

Described integrated chip also comprises at least one feeder that is connected with the output of each collection channel, is used for collecting and deriving product.

And, utilize existing micro/nano level process technology, through whirl coating, exposure, development, pour into a mould, peel off, step such as bonding, can produce based on dimethyl silicone polymer (Polydimethylsiloxane, PDMS), the micro-fluidic large scale integrated chip (LSI chip) of polymethyl methacrylate (PMMA) or poly-carbon ester (PC).

For example, H1 is 1, H2 to HN is 2, as shown in figure 11, the line of the corresponding collection channel input of the outlet of described special modality M with it, perpendicular to the line of two special modality N outlets, promptly each special modality M is corresponding to two special modality N, two special modality N outlets separately, the center line that both are linked to be line segment are the outlet of special modality M and the line of the input of corresponding collection channel; And generally, these two special modality N are symmetrical arranged about above-mentioned center line; That is to say, this is a special case, at this moment, two outlets can not form a definite circle, be still and be equally distributed on a circumference that pre-sets radius circular, therefore, two special modality N outlets separately, the perpendicular bisector that both are linked to be line segment is the outlet of special modality M and the line of the input of corresponding collection channel.Because it is not unique that two special modality N export the circle that is constituted, therefore aforementioned line also is to belong to one of the outlet of special modality M and the vertical line in the described circular center of circle.

Like this, when N is 9, M is 8 o'clock, final (M-1) power of 2 that forms of ground floor structure then, i.e. 27 powers, i.e. 128 channel outlet, final (N-1) power of 2, i.e. 28 powers, i.e. 256 channel outlet of forming of second layer structure; That is to say that it has 128 passage M, each passage M correspondence two passage N, has 128 outputs; For example passage M1 correspondence passage N11 and passage N12, and passage N11 is relative with the outbound course of passage N12, and is located along the same line; The distance of the outlet that exports to passage M1 of passage N11, identical with the distance of the outlet that exports to passage M1 of passage N12.

In above-mentioned example, the characteristic feature of device has been integrated 128 expansion shape nozzle chambers, for example, and as shown in Figure 3, ground floor structure input gas, as shown in Figure 2, second layer structure input liquid; Nozzle arrangements as shown in Figure 4, each gas output channel mouth correspondence two relative liquid output channel mouths; Then gas is at the narrowest point of nozzle, and promptly jet hole also is the narrowest point of collection channel simultaneously, has maximum shearing force, as shown in Figure 5, at expansion shape nozzle place, produces a decrease of speed gradient subsequently, is beneficial to the focusing of microvesicle and comes off; Therefore, utilize nozzle array can improve the production efficiency of microvesicle.

More particularly, in the part example of the present invention, adopted flow focusing principle (flow-focusing) to produce microvesicle, gas is in coaxial mobile center under the bag quilt of both sides liquid, it is the axis, gas forms stable cone by the fluid-focus of high-speed motion, and with the jet hole aligned in position, because the jet hole place is the narrowest, have maximum shearing force, as shown in Figure 5, expansion shape nozzle subsequently produces a velocity gradient, the microjet that is gone out by vertex of a cone end-fire comes off at the jet hole place, forms the microvesicle of single-size.

Each embodiment above adopting just can be used for micro-fluidic large scale integrated chip (LSI chip) the preparation of acoustic contrast agent, and the described integrated chip of the various embodiments described above can also be applied in other miniature mixtures of preparation, for example, and little drop or the like.

And, adopting the described integrated chip of above-mentioned arbitrary embodiment, the present invention also provides a kind of device.This device comprises the first material input block, the second material input block, storage unit and at least one as integrated chip as described in above-mentioned arbitrary embodiment, and these integrated chips form at least one integrated chip array.The feeder of each integrated chip is connected with described storage unit respectively, is used for collecting and deriving product; The described first material input block is connected with each passage 1 of one deck structure of each integrated chip respectively, is used to import first material; The described second material input block is connected from each passage 1 of another layer structure of each integrated chip respectively, is used to import second material.This device can be used for preparing micrometre level dispersoid, comprises acoustic contrast agent or other miniature mixtures.

Wherein, the combination of described first material input block and the described second material input block is the combination of gas input block and liquid input block; Be that the described first material input block is the gas input block, the described second material input block is the liquid input block, and perhaps, the described first material input block is the liquid input block, and the described second material input block is the gas input block.Perhaps, described first material input block and the described second material input block are the liquid input block.

For example, this device comprises gas input block, liquid input block, storage unit and at least one as integrated chip as described in above-mentioned arbitrary embodiment, and these integrated chips form at least one integrated chip array.In other words, this device comprises: be used to control the instrument of the flow parameter of gas, liquid, and be used for gas and the liquid chip by the micro-nozzle array, described micro-nozzle has consistent basically diameter.

For example, the block diagram of whole ultrasonic contrast preparation preparation facilities as shown in Figure 1, gas source output gas through flowmeter or pressure gauge control flow, outputs in the extensive micro-fluidic chip; The fluid supply output liquid through numerical control syringe pump control flow, outputs in the extensive micro-fluidic chip; Reaction generates product, carries out product and collects, and for example collects by the product collecting device.For example, by the feeder of each integrated chip, collect and the derivation product, these feeders are connected with described storage unit respectively, and final product is imported in the described storage unit; Can collect final product effectively like this.

An example is, described gas input block is connected with each passage 1 of one deck structure of each integrated chip respectively, is used to import gas; Described liquid input block is connected from each passage 1 of another layer structure of each integrated chip respectively, is used to import liquid.

For example, described gas input block is connected with each passage 1 of the ground floor structure of each integrated chip respectively, and described liquid input block is connected from each passage 1 of the second layer structure of each integrated chip respectively; Perhaps, described gas input block is connected with each passage 1 of the second layer structure of each integrated chip respectively, and described liquid input block is connected from each passage 1 of the ground floor structure of each integrated chip respectively.

An example is, described gas input block comprises pressure gas tank, pressure-reducing valve, first transfer tube, micro-flowmeter, pressure regulator valve and second transfer tube that is linked in sequence, and described second transfer tube is connected with each passage 1 of one deck structure of each integrated chip respectively.

Another example is, described liquid input block comprises reservoir, the 3rd transfer tube, digital control type syringe pump and the 4th transfer tube that is linked in sequence, and described the 4th transfer tube is connected with each passage 1 of another layer structure of each integrated chip respectively.

A preferred example is, in each integrated chip of described device, each channel cross-section is a rectangle, and it highly is 20 μ m to 30 μ m; The width of passage M cross section is that the width of 30 μ m to 50 μ m, passage N cross section is 50 μ m to 100 μ m, and perhaps, the width of passage M cross section is that the width of 50 μ m to 100 μ m, passage N cross section is 30 μ m to 50 μ m; Wherein, the structure of described collection channel comprise at least following structure one of them: export-oriented cone, spheroid, hemisphere or the cone table that amplifies, the caliber of its input is 7 μ m to 25 μ m, highly is 20 μ m to 30 μ m; Perhaps, cylinder or cuboid, its sectional area along the special modality M outbound course of correspondence is slightly less than this special modality M sectional area in the direction, highly is 20 μ m to 30 μ m.The matrix of described integrated chip is silicon, glass, dimethyl silicone polymer, polymethyl methacrylate or poly-carbon ester.The combination of ground floor structure and second layer structure is the combination of pathway structure and fluid passage structure.H1 is 1, and H2 to HN is 2; N is 8, M is 7, ground floor structure and second layer structure all are provided with two initial channel in the collection channel bilateral symmetry, i.e. 2 gas accesses and 2 liquid inlets, then the ground floor structure finally forms (M-1) power of 22, i.e. 128 channel outlet, final (N-1) power that forms 22 of second layer structure, i.e. 256 channel outlet; In this example, the characteristic feature of device has been integrated 128 expansion shape nozzle chambers, gas is at the narrowest point of nozzle, it is the narrowest point of collection channel, have maximum shearing force, expansion shape nozzle subsequently produces a decrease of speed gradient, is beneficial to the focusing of microvesicle and comes off; Nozzle array can improve the production efficiency of microvesicle.

More particularly, as shown in Figure 1, gas is transported to the gas access of chip through pressure-reducing valve by pressurized tank, by micro-flowmeter, pressure regulator valve monitoring gas flow, successional liquid pumps into the liquid inlet by the digital control type syringe pump with constant flow velocity, stored by special reservoir bottle through the microvesicle that micro-fluidic large scale integrated chip (LSI chip) produces, this reservoir bottle is a prior art.

Wherein, micro-fluidic large scale integrated chip (LSI chip) is to utilize micro-nano process technology to be made, and for example, this chip has double-layer structure up and down.

An example is more specifically, and this chip fabric as shown in Figure 2.Passage is symmetrical arranged the liquid inlet by two to begin to expand step by step, and finally through 8 grades of gradients, totally 7 grades of expansions form 256 branches, and promptly N is 8.Per two liquid inlets that belong to the branch of an initial inlet as micro-nozzle so just form 128 micro-nozzles.The position a ' of chain-dotted line indication is the gas access of a micro-nozzle among Fig. 2, totally 128 gas accesses, and the microvesicle of generation is collected storage through feeder.

This chip top layer channel design as shown in Figure 3, and is similar with fabric, and passage is symmetrical arranged the gas access by two to begin to expand step by step, and 7 gradients are arranged, and through 6 grades of expansions, promptly M is 7,64 branches of the final formation of single initial gas inlet.Therefore, formed 128 branches altogether, communicated, communicated with a ' as a with 128 gas accesses of the corresponding gas of bottom through expansion.

Wherein, the structure of bottom and top layer can be exchanged, as shown in Figure 6, this is the replacement form of micro-fluidic large scale integrated chip (LSI chip) among the present invention, top layer is the fluid passage structure, bottom is mainly gas passage, and two of bottom initial inlets are extended to 128 branches through 6 grades, as the gas access of micro-nozzle.Fig. 7 is the enlarged drawing of a micro-nozzle of part shown in the frame of broken lines among Fig. 6, is that two liquid inlets communicate with the top layer fluid passage with the main distinction of Fig. 4.

As shown in Figure 4, it is the enlarged drawing of part shown in Fig. 2 dotted line, schematic diagram for a micro-nozzle in this integrated chip array, micro-nozzle is the elementary cell that microvesicle forms, wherein, arrow is represented the flow direction of gas, liquid, and a gas access communicates with the top layer gas passage, in the narrowest place shearing force maximum of the collection channel of big arrowhead form.The bore of collection channel inlet is in 7 μ m-25 mu m ranges, channel height is 25 μ m, preferably with consistent with the height of other passages in one deck structure, the gas access width is 30 μ m-50 μ m, the liquid inlet width is 50 μ m-100 μ m, gas forms the stable taper that is positioned at the position, axis under the swiftly flowing fluid-focus effect in both sides, as shown in Figure 5, and with the jet hole aligned in position, because the jet hole place is the narrowest, has maximum shearing force, expansion shape nozzle subsequently produces a velocity gradient, and the microjet that is gone out by vertex of a cone end-fire comes off at the jet hole place, forms the microvesicle of single-size, microvesicle forms schematic diagram as shown in Figure 5, can see taper and microvesicle that flow focusing forms visually.

Wherein, as shown in Figure 9, the micro-nozzle shape of this device can be replaced by other geometry that forms velocity gradient, as rectangular configuration, only needs to produce a velocity gradient, and the microjet that allows vertex of a cone end-fire go out comes off at the jet hole place and gets final product.

Need to prove, amplify for portion size among each figure that this is for the purpose that illustrates, and do not draw according to the ratio of strictness.These illustrated not strict draftings all do not constitute any additional limits to each technical scheme of the present invention and each embodiment.

Each embodiment above adopting just can extensive integrated expansion shape micro-nozzle array, by importing gas and liquid at different feeder connections, can realize and help the large-scale production of microvesicle.Adopt various gas and liquid, import gas from each passage 1 of the ground floor structure of respectively organizing structure of integrated chip, device of the present invention can be used for preparing polytype acoustic contrast agent microvesicle.

Liquid is promptly imported in exchanging gas, liquid inlet from each passage 1 of the ground floor structure of respectively organizing structure of integrated chip, can be used for generating a large amount of little drops; Perhaps, feed two kinds of inconsistent liquid respectively, can generate little drop of bottom liquid parcel supernatant liquid, be highly suitable for food service industry and cosmetic industry.

Wherein, adopt the formed micro-nozzle array of a plurality of collection channels, the quantity of micro-nozzle can increase and decrease, for one group of structure, and the rarest nozzle, when the ground floor structure was provided with N level gradient, the expansion progression of passage was N-1, promptly counts N than gradient and lacks 1.Under the situation of having only an initial inlet, last branch is 2N-1, and the number of nozzle generally is 2N-2.Under the situation that two symmetrically arranged initial inlets are arranged, the number of nozzle is generally 2N-1.For example, N is 8 o'clock, and 8 grades of gradients are arranged, and has expanded 7 grades, expands one times for every grade, and first order gradient is provided with 1, the eight grade of gradient of 1 passage 128 passages 8 are set, under the situation of having only an initial inlet, to 64 nozzles should be arranged; Under the situation that two symmetrically arranged initial inlets are arranged, to 128 nozzles should be arranged.Be easy to control under the prerequisite of rate of flow of fluid, gas pressure, the nozzle number is The more the better.

And, a kind of method for preparing micrometre level dispersoid also is provided, immiscible between these micrometre level dispersoids; This method is applied to one and comprises in the device of integrated chip array, and described integrated chip array is made up of as integrated chip as described in above-mentioned arbitrary embodiment at least one; Described device is the described device of above-mentioned arbitrary embodiment.Said method comprising the steps of.

A1, pre-set condition by first, to each passage 1 input first material of one deck structure of each integrated chip; Pre-set condition by second, to each passage 1 input second material of another layer structure of each integrated chip.Wherein, first material can be different liquid with second material; Perhaps, first material can be liquid, and second material can be gas; Perhaps, first material can be gas, and second material can be liquid.

An example is, in the steps A 1, to each passage 1 input gas of one deck structure of each integrated chip, pressure is lower than 10psi, and pressure gets final product greater than 0, to each passage 1 input liquid of another layer structure of each integrated chip, flow velocity is lower than 3 μ L/s, and flow velocity gets final product greater than 0; For example, the pressure of input gas is lower than 5psi, and the flow velocity of input liquid is lower than 1.5 μ L/s; And for example, the pressure of input gas is lower than 8psi, and the flow velocity of input liquid is lower than 2.5 μ L/s; And for example, the pressure of input gas is lower than 7.5psi, and the flow velocity of input liquid is lower than 2.0 μ L/s; And for example, the pressure of input gas is lower than 4psi, and the flow velocity of input liquid is lower than 2.8 μ L/s.Like this, just can prepare the microvesicle of homogeneous, these microvesicles can be applicable in the acoustic contrast agent.

In concrete the application, described gas comprise at least nitrogen, fluorine carbon gas, fluorine sulphur class gas one of them; Described liquid comprise at least phospholipid liquid, surfactant class I liquid I, aforesaid liquid modification liquid one of them.

In order to guarantee that microvesicle has the long time-to-live in vivo, this installs spendable gas material mainly is fluorine carbon, the fluorine sulphur class gas that low solubility is arranged in blood plasma, as C3F8, C4F10, SF6 etc., also can be combination between these gas componants or they and the combination of nitrogen such as the combination of nitrogen and C4F10, nitrogen and C3F8.

And the preparation vacuole, i.e. microvesicle, its shell material is from liquid, and these liquid mainly comprise phospholipid, as DPPC, DPPA, DPPE, DSPC, DSPE, DSPA etc.; Surfactant-based, as Span20, Span60, Span 80 etc., Tween20, Tween 60, Tween 80 etc., and the combination of phospholipid, surfactant-based combination or phospholipid and surfactant-based combination; These materials can significantly reduce the surface tension of gas-liquid interface, have excellent biological compatibility and blood compatibility again.

Perhaps, the shell material of preparation vacuole is to carry out the material with specific function that modification obtains on the basis of above-mentioned shell material, comprises DSPE-PEG2000, DSPE-PEG5000, DSPE-PEG (2000) Biotin, DSPE-PEG (2000) Carboxylic Acid or the like.The hydrophobicity long-chain of phosphatide and the molecular structure of hydrophily polar head make it can adjust its molecular structure voluntarily in aqueous environments, hydrophobic chain closely is arranged in together, and hydrophilic head is exposed to water, the final vesica shape structure that forms, this structure can hinder the microvesicle that gas diffuses out formation, and hydrophilic shell makes it have good biocompatibility.Simultaneously, different polar heads can make phosphatide have different charges, by composition and the ratio of adjusting these charged phosphatide, can adjust the stability of phosphatide microvesicle.The phosphatide that contains the PEG long-chain as DSPE-PEG2000, has its circulation time ability of being caught by reticuloendothelial system in vivo of opposing, prolongs the time of circulation in vivo.The phosphatide that contains the Biotin part as DSPE-PEG (2000) Biotin, has identification antibody avidin ability, therefore has target function.Like this, by making up between the different phospholipid fractions, and adjust the ratio of each component, just can obtain the microbubble of difference in functionality and effect.

The initial conditions of A2, adjusting first material and/or second material, the material of each passage M output that makes each ground floor structure is with coaxial liquid form, by the material of each special modality N of correspondence by the axial material of parcel passage M output, form respectively and wait to collect product stream, it axially overlaps with described vertical line.

For example, in the steps A 2, regulate the initial conditions of gas with various or different liquids separately, the initial conditions of perhaps independent adjustments of gas or liquid, the perhaps initial conditions of adjustments of gas and liquid simultaneously, the material of each passage M output that makes each ground floor structure be with coaxial liquid form, by the material of each special modality N of correspondence by the material that axially wraps up respective channel M output, form respectively and wait to collect product stream, it axially overlaps with described vertical line; For example, what form a reverse taper respectively waits that collecting product flows, and it axially overlaps with described vertical line.

A3, collect and derive product from the output of each collection channel.

For example, to each passage 1 input gas of the ground floor structure of each integrated chip, to each passage 1 input liquid of the second layer structure of each integrated chip; Like this, can obtain a kind ofly focusing on the method that the nozzle place forms microvesicle by the both sides liquid flow, form highly monodispersed microvesicle, be suitable for the ultrasonic contrast imaging technology by gas.Perhaps, to each passage 1 input liquid of the ground floor structure of each integrated chip, to each passage 1 input gas of the second layer structure of each integrated chip; Like this, can obtain a kind ofly focusing on the method that the nozzle place forms little drop by the both sides gas flow by liquid.

Same as above, a preferred example is, described integrated chip, and H1 is 1, and H2 to HN is 2, and N is 9, and M is 8.It has 128 passage M, and each passage M correspondence two passage N, has 128 outputs; For example passage M1 correspondence passage N11 and passage N12, and passage N11 is relative with the outbound course of passage N12, and is located along the same line; The distance of the outlet that exports to passage M1 of passage N11, identical with the distance of the outlet that exports to passage M1 of passage N12.

Another example is, in the steps A 1, to each passage 1 input gas of the ground floor structure of each integrated chip, to each passage 1 input liquid of the second layer structure of each integrated chip; In the steps A 2, the gas of each passage M output that makes each ground floor structure is with coaxial liquid form, by each passage N of the corresponding a certain passage M of each second layer structure, the liquid that is flowed out, forms respectively and waits to collect microballoon stream gas that should passage M by axial parcel.Wherein, coaxial liquid form as described above.

On the basis of last example, in the steps A 2, the gas of each passage M output of each ground floor structure forms the stable reverse taper that the axis is positioned at described vertical line position respectively under the swiftly flowing fluid-focus effect in both sides, the tip of each reverse taper is relative with each collection channel position respectively.

Another example is, in the steps A 1, to each passage 1 input first liquid of the ground floor structure of each integrated chip, to each passage 1 input gas or second liquid of the second layer structure of each integrated chip; In the steps A 2, first liquid of each passage M output that makes each ground floor structure is with coaxial liquid form,, formed respectively and waits to collect little stream of liquid droplets by axial parcel by gas or second liquid; What for example form a reverse taper respectively waits to collect little stream of liquid droplets.

In above-mentioned each example, in the steps A 1, described liquid can add ligands specific in advance; Perhaps, after the steps A 3, add ligands specific to described product.Like this, can add different ligands specifics, thereby prepare targeted ultrasound contrast agent as required.

Continue below integrated chip, apparatus and method are especially prepared the method for acoustic contrast agent, make a detailed description.

In the ultrasonic contrast imaging technology, it reaches the size impact of microbubble contrast agent ultrasonic reflectivity by the microcirculatory ability of lung, diameter must just can not cause obstruction by lung's arteriole safely less than 7 μ m, wherein, ultrasonic scattering strength, incident intensity have following relation formula (1):

$I/I_0~\frac{1}{9}\mathrm{nV}[k^4{r}^6{({\mathrm{\γ}}_c+{\mathrm{\γ}}_d\cos \mathrm{\θ})}^2/d^2]---\left(1\right)$

Wherein, I, I ₀Be respectively ultrasonic scattering strength, incident intensity; N is the scattering particles number; V is a scattering volume; K is a wave number; R is particle radii, i.e. the microvesicle radius; γ _cBe a compression (γ _c=(k _s-k _m)/k _m, k _s, k _mBe respectively the compression ratio of scattering particles and the compression ratio of medium, known technology); γ _dBe density item (γ _d=(3 ρ _s-3 ρ _m)/(2 ρ _s+ ρ _m), ρ _s, ρ _mBe respectively scattering particles and density of medium, known technology); θ is an angle of scattering; D is the distance to scattering particles.By formula (1) as can be seen, the scattered power of microvesicle is a microvesicle radius cubo-cubic function, promptly, the scattering strength of acoustic contrast agent microvesicle is directly proportional with bubble radius six powers, illustrate that microvesicle is more little, reflectivity is poor more, so the optimum diameter size of microvesicle can not be too little.Therefore, clinical practice requires the optimum diameter size of microvesicle between 2 μ m to 5 μ m.

Main relevant with the flow rate of gas, liquid by the microvesicle diameter that micro-fluidic focusing system produces, capillary influence is less, can ignore, shown in following formula (2):

d _b/D∝(Q _g/Q _l) ^0.4 (2)

Wherein, Q _g, Q _lIt is respectively the flow rate of gas, liquid; D is the jet hole diameter; Q _g/ Q _l＜1.

Utilize pressure regulator valve to adjust gas pressure, the flow rate of syringe pump flow parameters control liquid is set, (Pounds per square inch, 1psi=6.895kPa), flow rate of liquid Q＜1.5 μ L/s can obtain diameter d to gas pressure P＜5psi generally speaking _bThe microvesicle of＜5 μ m, the polydispersity index of microvesicle particle diameter＜2%.

In order to confirm the particle diameter and the production efficiency of the microvesicle that apparatus of the present invention prepare, adopt an above-mentioned integrated chip to carry out following experiment, it is two initial inlets of symmetry in each layer structure, H1 is 1, H2 to HN is 2, and N is 8, and M is 7, the ground floor structure has 128 M passages, and second layer structure has 256 N passages; It is integrated 128 expansion shape nozzle chambers, the ground floor structure feeds gas, in the narrowest some output of nozzle, as Fig. 4, shown in Figure 5.Specify as follows.

Experiment one:

Gas adopts nitrogen, and liquid adopts PBS (PBS) 8ml of following preparation: PH=7.4, and Tween 80 (Tween80) 1ml is mixed with uniform mixture, and adjustments of gas pressure is 1.8psi, liquid flow rate 2.0 μ Ls ^-1, can obtain the microvesicle diameter and be about 3.6 μ m, per minute approximately can produce 7 * 109 microvesicles.As shown in Figure 8, be the picture of the microvesicle of the generation under the Olympus inverted microscope, photographed, the microvesicle particle diameter distributes and has high degree of monodispersity as can be seen, and particle diameter distributes more even, and outer casing thickness person is more even.

Experiment two: on the basis of experiment one, liquid flow rate is constant, increase gas pressure to 4.5psi, obtain the microvesicle diameter and be about 6 μ m, the result who obtains from microscope, particle diameter distributes same relatively more even, and comparative experiments one and experiment two can obtain: change the size that gas pressure can be controlled the microvesicle particle diameter flexibly; In like manner, also flow rate of liquid can be changed certainly, identical effect can be reached.

Experiment three: on the basis of experiment one, gas pressure is constant, increases flow rate of liquid to 2.7 μ Ls ^-1, obtaining the microvesicle diameter and be about 3 μ m, particle diameter distributes same relatively more even, and comparative experiments two and experiment three can obtain: can change the size that flow rate of liquid can be controlled the microvesicle particle diameter flexibly.

Experiment four: gas adopts perfluorocarbon (PFC), liquid adopts following method preparation: lipid two palmitic acid phosphatid ylcholines (DPPC), two palm phosphatidic acid polyethylene glycol (DPPA), 5000 change two palmitic acid phosphatidyl-ethanolamines (DPPE-PEG5000) with 81: 8: 10 mol ratio, be dissolved in the chloroform (CHCl3), under vacuum and condition of nitrogen gas, form uniform mixture, the fluorescer (DiI-C18) of adding 1mol% and the NaCl solution of 4mg/ml are in the test tube that contains mixture of phospholipids, ultrasonic processing 20 minutes and sneak into the glycerine and 1 of 10% concentration under the room temperature, the mixture of 2 propane diols (GPW), gas pressure 10psi, liquid flow rate 1.0 μ Ls ^-1, can obtain microvesicle diameter 5 μ m by said chip, per minute approximately can produce 8 * 109 microvesicles.

The present invention has the following advantages:

The most significant advantage is that the acoustic contrast agent microvesicle of making has high degree of monodispersity and particle diameter controllability, satisfies the requirement of ultrasonic contrast imaging technology.The polydispersity index of microvesicle particle diameter＜2%, particle diameter become along with the increase of gas pressure greatly, reduce along with the increase of flow rate of liquid, control very flexible.And,, improved the preparation efficiency of microvesicle greatly by adopting the micro-nozzle array; Simultaneously, this device has reusability, has reduced production cost.

Device of the present invention can be used for preparing polytype acoustic contrast agent microvesicle, and the microvesicle outer casing thickness of preparation is even.

Conventional acoustic contrast agent can be directly with this device preparation, as lipid, albumin class, polymer class, surfactant-based etc.; As mentioned above, the preparation of targeted ultrasound contrast agent has two kinds of selections, and a kind of is at present the most frequently used: add ligands specific after conventional acoustic contrast agent preparation is finished, preparation separately; Another kind is to add ligands specific in advance in the preparation liquid before contrast preparation preparation is finished, ligands specific will be mounted on the shell of microvesicle when forming contrast agent microbubble, some shell material such as protein molecular inactivation under high temperature and ultrasound condition, thus this method be in the past sound shake method can not.This contrive equipment all is suitable for these two kinds of methods, particularly has more advantage when adopting a kind of selection in back, has not only reduced the preparation link, has also reduced the destruction of preparation link microvesicle.

And the heat production in the contrast preparation preparation process of apparatus of the present invention and method is few, therefore especially is suitable for preparing the acoustic contrast agent of double medicine or gene targeting vector.Contrast agent microbubble is carried the mode of gene or medicine and is divided two kinds: stick method and integration method, wherein integration method has remarkable advantages, reason is: stick the surface that method only is attached on medicine or gene by simple mixing contrast agent microbubble, binding capacity is few on the one hand, medicine that sticks on the other hand or gene combine insecure with microvesicle, after intravenous injection, under impacting, blood flow is easy to come off, and target is poor; And integration method both can be attached on the microvesicle surface with medicine or gene, more it can be incorporated on the microvesicle film or be wrapped in the microvesicle, when binding capacity is increased, make target better again.When employing sound shakes method usually, in preparation process, produce higher temperature, can not add medicine or gene simultaneously in the preparation, can only adopt and stick the method preparation, apparatus of the present invention heat production in preparation process is few, therefore can adopt the acoustic contrast agent of double medicine of integration method preparation or gene targeting vector.

Easy, effective and safety that this acoustic contrast agent prepares has very high using value.Simultaneously, as mentioned above, the present invention also can be used for preparing little drop.

At a gas access; the simplest microvesicle of two liquid inlet formations prepares the unit; its throughput rate is about 107/minute; extensive micro-fluidic chip can significantly improve productive rate described in this patent; prepare the unit as integrated 128 microvesicles; then its productive rate can be brought up to 109/minute; in addition; compare with existing commercialization acoustic contrast agent; a kind of acoustic contrast agent Difinity as U.S. food and the clinical use of FAD (FDA) approval; its productive rate is about 109/45 second; the productive rate of the described extensive micro-fluidic chip of this patent can compare favourably with it; the more important thing is; the particle diameter of Difinity contrast preparation distributes and has polydispersity; the particle diameter wider distribution; average diameter is about 1.8 μ m; the standard deviation of diameter is 1.5 μ m; but maximum microvesicle diameter reaches 20 μ m unexpectedly; thereby cause the resonant frequency scope of microvesicle bigger; and the restriction of the bandwidth of present ultrasonic image-forming system can only detect very narrow resonant frequency scope; so approximately have only 18% microbubble signals to detect; in other words; 82% microvesicle is inoperative and slattern; reduced the sensitivity of imaging system; the contrast preparation of the described device preparation of this patent has very high monodispersity; microvesicle ratio with useful signal is higher than 92%; can improve the sensitivity of ultrasonic contrast imaging technology; particularly in ultrasonic molecular image technology; the adherence rate of targeted contrast agent is about every square millimeter of 5-25; the monodispersity of microvesicle seems particularly important to improving imaging sensitivity; therefore; say that in this sense the contrast preparation of the described device preparation of this patent has absolute advantage aspect large-scale production, clinical value is very high.

Should be understood that, for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.

Claims (19)

1, a kind of integrated chip is characterized in that, is one group of structure with two-layer adjacent structure, and described integrated chip comprises at least one group of structure;

In one group of structure, the ground floor structure is provided with M level gradient, and second layer structure is provided with N level gradient; Wherein, M, N are natural numbers, and M is less than N; In the gradients at different levels, first order gradient is provided with H1 passage 1, and second level gradient is drawn H2 branch from each passage 1, form H1 * H2 passage 2, by that analogy, N level gradient is drawn HN branch from each passage (N-1), forms H1 * H2...... * HN passage N;

In one group of structure,, all formed H (M+1) * H (M+2) from what a passage M drew in N level gradient ... the special modality N of * H (N-1) * HN quantity corresponding to second layer structure for arbitrary special modality M of ground floor structure;

Wherein,

The outlet of each special modality N is equally distributed on a circumference that pre-sets radius circular;

For this special modality M, its outbound course is perpendicular to this circle, and its outlet is passed through this circular center of circle with this circular vertical line, and the outlet of this special modality M and this distance of center circle are from a length-specific;

The outbound course of each special modality N is identical acute angle with the angle of described vertical line;

Along the outbound course of this special modality M,, a collection channel is set in the side of this circle away from this special modality M;

Described integrated chip also comprises at least one feeder that is connected with the output of each collection channel, is used for collecting and deriving product.

2, according to the described integrated chip of claim 1, it is characterized in that M=N-1.

According to the described integrated chip of claim 1, it is characterized in that 3, the outbound course of each special modality N is by this circle center of circle.

According to the described integrated chip of claim 1, it is characterized in that 4, the cross section of passage M and passage N is a rectangle; Described rectangle height is that 20 μ m to 30 μ m, width are respectively 30 μ m to 50 μ m, 50 μ m to 100 μ m, perhaps, is respectively 50 μ m to 100 μ m, 30 μ m to 50 μ m.

5, according to the described integrated chip of claim 1, it is characterized in that, the structure of described collection channel comprise at least following structure one of them: export-oriented cone, spheroid, hemisphere or the cone table that amplifies, the caliber of its input is 7 μ m to 25 μ m, highly is 20 μ m to 30 μ m; Perhaps, cylinder or cuboid, its sectional area along the special modality M outbound course of correspondence is slightly less than this special modality M sectional area in the direction, highly is 20 μ m to 30 μ m.

According to the described integrated chip of claim 1, it is characterized in that 6, its matrix is surface hydrophilicity matrix.

According to the described integrated chip of claim 6, it is characterized in that 7, its matrix is silicon, glass, dimethyl silicone polymer, polymethyl methacrylate or poly-carbon ester.

According to the described integrated chip of claim 1, it is characterized in that 8, the combination of ground floor structure and second layer structure is the combination of pathway structure and fluid passage structure.

9, a kind of device is characterized in that, comprising: the first material input block, the second material input block, at least one as claim 1 to claim 8 arbitrary as described in integrated chip form integrated chip array, storage unit;

The feeder of each integrated chip is connected with described storage unit respectively, is used for collecting and deriving product;

The described first material input block is connected with each passage 1 of one deck structure of each integrated chip respectively, is used to import first material;

The described second material input block is connected from each passage 1 of another layer structure of each integrated chip respectively, is used to import second material.

According to the described device of claim 9, it is characterized in that 10, the combination of described first material input block and the described second material input block is the combination of gas input block and liquid input block; Perhaps, described first material input block and the described second material input block are the liquid input block.

11, according to the described device of claim 10, it is characterized in that, described gas input block comprises pressure gas tank, pressure-reducing valve, first transfer tube, micro-flowmeter, pressure regulator valve and second transfer tube that is linked in sequence, and described second transfer tube is connected with each passage 1 of one deck structure of each integrated chip respectively;

Described liquid input block comprises reservoir, the 3rd transfer tube, digital control type syringe pump and the 4th transfer tube that is linked in sequence, and described the 4th transfer tube is connected with each passage 1 of another layer structure of each integrated chip respectively.

12, a kind of method for preparing micrometre level dispersoid is characterized in that, be applied to one and comprise in the device of integrated chip array, described integrated chip array by at least one as claim 1 to claim 8 arbitrary as described in integrated chip form;

Said method comprising the steps of:

A1, pre-set condition by first, to each passage 1 input first material of one deck structure of each integrated chip; Pre-set condition by second, to each passage 1 input second material of another layer structure of each integrated chip;

The initial conditions of A2, adjusting first material and/or second material, the material of each passage M output that makes each ground floor structure is with coaxial liquid form,, formed respectively and wait to collect product stream by axially wrapping up by the material of each special modality N of correspondence, it axially overlaps with described vertical line;

A3, collect and derive product from the output of each collection channel.

According to the described method of claim 12, it is characterized in that 13, the combination of described first material and described second material is the combination of gas and liquid; Described gas comprise at least nitrogen, fluorine carbon gas, fluorine sulphur class gas one of them; Described liquid comprise at least phospholipid liquid, surfactant class I liquid I, aforesaid liquid modification liquid one of them.

14, according to the described method of claim 12, it is characterized in that, in the steps A 1, to each passage 1 input first liquid of the ground floor structure of each integrated chip, to each passage 1 input gas or second liquid of the second layer structure of each integrated chip;

In the steps A 2, first liquid of each passage M output that makes each ground floor structure is with coaxial liquid form,, formed respectively and waits to collect little stream of liquid droplets by axial described first liquid of parcel by gas or second liquid.

According to the described method of claim 12, it is characterized in that 15, in the steps A 1, to each passage 1 input gas of one deck structure of each integrated chip, pressure is lower than 10psi, to each passage 1 input liquid of another layer structure of each integrated chip, flow velocity is lower than 3 μ L/s; In the steps A 2, the initial conditions of adjustments of gas and/or liquid.

According to the described method of claim 15, it is characterized in that 16, in the steps A 1, the pressure of input gas is lower than 5psi, the flow velocity of input liquid is lower than 1.5 μ L/s.

According to the described method of claim 15, it is characterized in that 17, in the steps A 1, described liquid adds ligands specific in advance; Perhaps, after the steps A 3, add ligands specific to described product.

18, according to the described method of claim 15, it is characterized in that, in the steps A 1, to each passage 1 input gas of the ground floor structure of each integrated chip, to each passage 1 input liquid of the second layer structure of each integrated chip;

In the steps A 2, the initial conditions of adjustments of gas and/or liquid, the gas of each passage M output that makes each ground floor structure be with coaxial liquid form, by axial parcel liquid, forms respectively and wait to collect microballoon stream.

19, according to the described method of claim 18, it is characterized in that, in the steps A 2, the gas of each passage M output of each ground floor structure, form the stable reverse taper that the axis is positioned at described vertical line position respectively under the swiftly flowing fluid-focus effect in both sides, the tip of each reverse taper is relative with each collection channel position respectively.

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