CN101479041A - A device and a method for promoting crystallisation - Google Patents

Abstract

The invention relates to a microfluidic device for promoting crystallisation of target molecules, such as proteins. The device comprises a solid structure with a top face and an opposite bottom face and with a least one liquid channel. The liquid channel comprises a target molecule solution inlet and at least two precipitant inlets. The target molecule solution inlet is in liquid communication with each of the precipitant inlets through the liquid channel. The liquid channel comprises a branching channel section adjacent to the target molecule solution inlet, crystallisation channel sections adjacent to the respective precipitant inlets and flow break channel sections arranged between the branching channel section and each of the crystallization channel sections. The liquid channel is branched from 1 to X in the branching channel section, wherein X is the number of crystallisation channel sections, and the flow break channel sections comprise a flow break arrangement capable of breaking up the liquid communication between said respective branching channel section and crystallisation channel sections.

Description

Promote the device and method of crystallization

Technical field

The present invention relates to be used to promote the microfluidic device of target molecule crystalline growth, relate to the microfluidic device that is used to promote as the big molecule growth of protein, nucleic acid and/or carbohydrate and so on especially.The invention still further relates to and utilize described device to promote the method for crystallization.The invention still further relates to and be used for the macromolecular microfluidic device of simple collection crystallization.

Background technology

Molecule such as macromolecular crystallization are the very important technology of biochemical field.Biochemical molecular such as nucleic acid, protein and carbohydrate have uncertain crystalline texture, and the 3D structure of molecules often plays an important role to their biological function.In order to obtain the detailed knowledge of the protein involved mode of action, key is to measure the three-dimensional structure of protein, because its 3D structure and function are closely related very much.When bioprocess technology need skillfully be operated, this 3D structure was just particularly useful, and it is found in the medical research.Now, surpassing 90% medicine on the market is little part with protein interaction.In order to understand this interaction and in creating new and improved medicine, to utilize it, must measure the 3D structure of part-protein complex.

The crystallization of molecule, the crystallization of for example big molecule such as protein is to make this target become to be difficult to the solid form that dissolves and turn back to its crystalline state to carry out by the chemical environment that target compound solution is provided and changes the target compound of dissolving.The precipitating reagent that the change of this chemical environment normally makes target compound be difficult to dissolve by introducing is finished.

US 6409832 has described a kind of microfluidic device that utilizes liquid-liquid to spread to promote protein crystal growth.This device comprises one or more mixed crystallizations chamber, and each chamber comprises the inlet of the inlet of protein solution and relative crystallizing agent (precipitating reagent).Protein solution enters the mouth by protein and introduces the filled chamber of designated volume, and enters mixing chamber from this.Crystallizing agent is packed into mixing chamber by the crystallizing agent inlet, and utilizes liquid-liquid diffusion that liquid is mixed.

US 2003/61687 discloses the different microfluid system that is used for the crystallization of high yield examination target material.Crystallization is by finishing in a plurality of chambers that target material solution are incorporated into little assembling fluid means.Operate little assembling fluid means then with the solution condition in the change chamber, thereby a large amount of crystalline environment is provided simultaneously.This microfluidics systems comprises the elastomer block that comprises little assembly room.

The system that is used for the target material crystallization comprises elastomer block, and this elastomer block comprises and is configured as the little assembly room that comprises a large amount of target material solution, and the little assembling fluid passage that is communicated with this chamber fluid, and introduce a large amount of crystallizing agents in the chamber this fluid passage.By with elastomeric material distortion to stop up fluid and flow or the mediation fluid flows or measures a certain amount of fluid and operate this structure to change the solution condition in the chamber.

Crystal system also can comprise isolation structure, this structure that is shaped optionally chamber and fluid passage isolates by being out of shape elastomeric wall when receiving the crystallizing agent of certain volume along with the fluid passage, and the chamber is contacted with the fluid passage with the solution condition in the change chamber.Alternatively, this crystal system can also comprise the control channel that covers on the chamber and separate by film with the chamber, and described film can be offset the sample solution that gets rid of calibrator quantity from the chamber in the chamber, makes film lax order about in the crystallizing agent inlet chamber of calibrator quantity.Further alternatively, crystal system can comprise a plurality of first horizontal fluid passage that are communicated with the target material fluid, and a plurality of and first fluid passage quadrature and intersect to form second horizontal fluid passage in many crosspoints with it, this second fluid passage is communicated with the crystallizing agent fluid to make and can form a series of solution environmentals at the place, crosspoint.

WO2004/067174 discloses the equipment that is used to carry out the microcosmic chemistry.This equipment comprises the permeable micro-fluid chip channel design of the steam with feed line.This chip structure has the first and second relative ends, and it can make first and second fluent materials flow toward each other from the opposite end of feed line by them and interact.This chip structure also has the valve mechanism in this structure, this valve mechanism operationally opens and closes feed line in the centre position between its first end and second end, thereby this chip structure can be mobile in proper order from occupied state and interacting state, in described occupied state, close the centre position so that fluent material blind filling (blind-filled) in the feed line of centre position opposite side, in described interacting state, open the centre position so that fluent material interacts.This equipment also comprises vaporizer, and this vaporizer is used to form the steam ambient around the chip structure, with the evaporation of fluent material in the feed line of compensation interacting state chip structure.

US 2005/0205005 discloses the microfluidic structures that is used for the high throughput screening crystallization of protein.This microfluidic structures comprises the hybrid chip of permission accurate measurement reagent with a large amount of potential crystallizations of rapid formation.This microfluidic structures is made by microcosmic machining elastomeric material, and has the fluid passage of the several layers that valve arrangement is provided.

The object of the present invention is to provide a kind of device that promotes the target molecule crystallization, this device is simple, production cost is low and be convenient to operation.

The object of the invention also is to provide a kind of straightforward procedure of operating this device.

The present invention has realized these purposes, limiting in claim.As hereinafter explaining, the present invention has further shown the beneficial property of comparing with method with the prior art crystallization apparatus with embodiment of the present invention.

Disclosure of the Invention

Compare with prior-art devices, of the present inventionly be used to promote that the device of target molecule crystallization is very economical, the production cost that its basic reason is this device is low and very simple, quick and reliable in the use simultaneously.

Term " promotes the target molecule crystallization " and comprises the further growth that forms initial crystal (embryos) and crystal.For some tests, expectation check small crystals.And for other test, can expect to allow crystal growth to go on up to forming bigger crystal.

Microfluidic device of the present invention comprises solid structure, and this solid structure has end face and opposed bottom surface and at least one fluid passage.This end face can be for example smooth substantially and optional parallel with the bottom surface, so that for example artificial or robot manipulation.Described fluid passage comprises target molecule solution inlet and at least two precipitating reagent inlets, and wherein target molecule solution inlet is by fluid passage and each precipitating reagent inlet fluid connection.The target molecule inlet opens wide or is come-at-able at least from the outside, in order to fill target molecule solution to fluid passage, for example utilizes pin by film or by passing the outlet of solid structure, the example of the end face of describing in for example following preferred embodiment.

Fluid passage comprises the branching channel section of contiguous described target molecule solution inlet, the crystallization channel section of the contiguous described inlet of precipitating reagent separately and be arranged on described branching channel section and described each crystallization channel section between cutout passage (flow break channel) section.

In one embodiment, microfluidic device can also comprise following further describe be arranged on cutout channel section between crystallization channel section and the precipitating reagent inlet.

In the branching channel section, fluid passage branches into 1-X, and wherein X is the quantity of crystallization channel section.X can be arbitrary integer in principle.In one embodiment, X is 2-1000, for example 2-400,4-100,8-48,8-24.

The cutout channel section comprises the cutout setting that can disconnect the fluid connection between described channel section of branching separately and the crystallization channel section.

Therefore, may carry out a plurality of crystallization trials, only target molecule solution be added a target molecule solution inlet simultaneously by this device.Target molecule solution can flow into many crystallization channel sections, and target molecule solution can touch the precipitating reagent from the precipitating reagent inlet herein.Simultaneously, any fluid between various crystallization channel sections is communicated with and can disconnects at the branching channel section, and described branching channel section is arranged between branching channel section and the crystallization channel section.Therefore, provide a kind of very simply device, its simple to operate and wherein mass crystallization test can carry out simultaneously.

In principle, cutout is provided with can be provided by any structure that the fluid that can disconnect effectively between various crystallization channel sections is communicated with.In fact, have been found that effective cutout setting can or be the form of solid structure difference that described cutout channel section is provided for the form of the difference between the channel section that stops separately, described cutout channel section is selected from a) branching channel section, and b) with the contiguous crystallization channel section of the described channel section that stops separately at least one.

Preferably, cutout is set at least a in the following form: a) capillary cutout, at least one cutout channel section wherein is set provides capillary force with the liquid in described passage, this capillary force is less than the capillary force that is provided to liquid by described contiguous crystallization channel section, b) passage displacement (channeldisplacement), wherein at least a portion cutout passage is with respect at least one vertical displacement in branching channel section and the crystallization channel section, and c) line of weakness (weakening line) cutout, wherein said solid structure be equipped with at least one cutout channel section in the fluid passage line of weakness of intersecting.

The vertical displacement of channel part is meant and is moved to respect to the visible higher or lower position of device end face.

In fact, when the device of designing optimal, particularly for comprise according to above a) or b) during the embodiment of cutout channel section of operation, the character of target molecule solution may be important.This be because the character of target molecule solution particularly surface tension whether can to stand capillary pulling power for solution be conclusive.When design during microfluidic device, can water as the substitute of target molecule solution, even use, when utilizing specific target molecule solution, can further optimize microfluidic device for some.As the following stated, the aqueous solution that target molecule solution can be preferably the aqueous solution and for example contain surfactant, this surfactant can change itself and the surface tension of water ratio.Yet those skilled in the art can utilize its ordinary skill to adjust design parameter.

In an embodiment of microfluidic device, the liquid of at least one cutout channel section in described passage is set capillary force is provided, the capillary force that this capillary force provides to liquid less than described contiguous crystallization channel section, described cutout channel section are equipped with by the capillary of its geometry and/or its surface tension setting arrheas (capillary flow stop).

When liquid preferably flowed into microfluidic channels with certain speed under capillary force, this flowed with less capillary force or does not have and continues one section under the situation of capillary force and do not quickened fully up to liquid.In device, this effect is used in this embodiment.Various sections of passage are set, make when target molecule solution flows into the branching channel section, this solution will continue to flow into the cutout channel section, even the capillary force that this cutout channel section that is provided with provides to liquid is less than the capillary force that is provided to liquid by vicinity crystallization channel section.Should flow lasts till that target molecule solution enters the crystallization channel section, thereby wherein said target molecule solution stands capillary force and fills the crystallization channel section.When flowing owing to the crystallization channel section is filled when stopping, this target molecule solution will can not flow back into the cutout channel section, because the capillary force that the cutout channel section that is provided with provides to target molecule solution is less than the capillary force that also has preferred branching channel section to provide to liquid by contiguous crystallization channel section.At last, the cutout channel section will drain, and connects and will disconnect at the liquid between the crystallization channel section separately.

The cutout channel section can be set to provide capillary force to liquid, this capillary force is less than the contiguous capillary force that the crystallization channel section/the branching channel section provides to liquid by any setting.In the microfluidic device field, known many methods that are used to be provided with the capillary performance of expectation, and those skilled in the art can use the setting of any appropriate.

In one embodiment, the wall that wherein fluid passage comprises around passage and passage length extends.In at least one cutout channel section, in part around it at least, described wall has the little surface tension of surface tension than the adjacent wall surface in contiguous crystallization channel section, thereby so that cutout setting to be provided.In this embodiment, preferably at least one cutout channel section, in major part around it at least, for example at least 60%, 80%, 90%, at least basically the whole surface around it has the little surface tension of surface tension than the adjacent wall surface in contiguous crystallization channel section, thereby so that cutout setting to be provided.

In one embodiment, in at least one cutout channel section, in part around it at least, wall (for example has obviously lower surface tension, utilize the contact angle measurement to measure), at least 5 dynes per centimeter for example, at least 10 dynes per centimeter, at least 20 dynes per centimeter, this surface tension is less than the surface tension of the adjacent wall surface in contiguous crystallization channel section and contiguous branching channel section at least one, preferably at least one cutout channel section, in part around it at least, wall has less than 73 dynes per centimeter, for example 60 dynes per centimeter, for example surface tension of 10-55 dynes per centimeter.

Surface tension can, for example utilize contact angle to measure.Less than for the surface of 73 dynes per centimeter, the contact angle of water/sample is determined as at least 90 degree in 20 ℃ of air for surface tension.Unless mention other, all mensuration is all carried out in 20 ℃ of air.

Surface energy and surface tension relate to two terms of the same nature on surface, and these two terms exchanges are used usually.The surface on surface can utilize tensiometer to measure, for example the rotation decline image tensiometer of SVT20, DataPhysics Instruments GmbH sale.In this was used, term " surface tension " was the surface energy of macroscopic view, and promptly it directly is proportional to the water-wet behavior on surface, and it can for example be measured by the contact angle of water droplet well known to those skilled in the art.In comparing and measuring, for example in measuring two surface portions which have the highest surface can the time, know that accurate surface can be unnecessary, simply relatively in two surfaces which have may be just enough to the less contact angle of water.

In one embodiment, the fluid passage that comprises the cutout part is made by non-elastomeric material.Therefore the shape of passage and size keep reliable.In addition, should preferably avoid elastomeric valve mechanism, because such elastomer valve door is difficult to operation and unreliable usually.In addition, shown that closing the elastomer valve by the fluid passage that compresses contiguous crystallization passage can disturb precipitating reagent to the target molecule Solution Diffusion, thereby and disturbed macromolecular crystallization.Therefore it is incompressible to expect that the cutout channel section comes down to, and the channel section that promptly stops can not be by elastic compression, and the channel section that preferably stops can not be come closing passage by elastic compression.

In order to be based upon the capillary flow of particular fluid in the fluid passage, the surface of at least some fluidic channel walls need have can order about the surface energy that liquid advances.According to a famous theory, however its scope that should not be construed as limiting the invention, if only the surface of at least some fluidic channel walls has less than the liquid contact angles of being discussed of 90 degree and just may set up capillary flow.In principle, angle is more little, and mobile meeting is fast more.Aspect this, can also mention according to the Young's equation ambient air and also can influence contact angle between liquid and the fluidic channel walls, this equation connects the surface tension of the liquid-steam surface tension force of contact angle, drop and the solid that contacts with liquid.

Measurement of contact angle is used as the objective of a kind of comparison surface tension force (comparativesurface tension) of measuring solid and simple method.Young's equation is represented is that the surface tension of solid directly is proportional to contact angle.This equation is:

g(sv)＝g(lv)(cosq)+g(sl)，

Wherein, g (sv) is the surface tension at solid-steam interface, and g (lv) is the interfacial surface tension of liquid-vapour interface, and g (sl) is the surface tension at interface between solid and liquid, and (q) is contact angle.

In theory, passage can have cross sectional shape arbitrarily, for example the polygon on ellipse, half elliptic, four limits (quadrilateral polygonal), square, rectangle and trapezoidal.Cross sectional shape can change or can be along its length direction unanimity.For example, a channel section can have first cross section, and another section of same fluid passage can have another cross sectional shape.

In an embodiment of microfluidic device, wherein said fluid passage has polygon (quadrilateral polygonal) cross sectional shape on four limits at least at its cutout channel section, the polygonal cross-section shape on this four limit provides bottom surface, relative end face and two sides, preferably in side and bottom surface and the end face has less than 73 dynes per centimeter (contact angle of water/sample is 90 degree) at least at least, for example less than the surface tension of 60 dynes per centimeter, 10-55 dynes per centimeter.

In an embodiment of microfluidic device, the wall that wherein said fluid passage comprises around passage and passage length extends.In in branching channel section and crystallization channel section at least one, at least a portion around it, described wall has at least 60 dynes per centimeter, for example the surface tension of at least 73 dynes per centimeter, at least 75 dynes per centimeter, at least 80 dynes per centimeter.In one embodiment, at least one in branching channel section and crystallization channel section, at least a portion around it, wall has the contact angle less than the water of 90 degree or preferred target molecule solution.

In an embodiment of microfluidic device, wherein fluid passage comprises around the passage and the wall of passage length extension.In at least one of branching channel section and crystallization channel section, described fluid passage has polygon (quadrilateral polygonal) cross sectional shape on four limits, the polygonal cross-section shape on this four limit provides bottom surface, relative end face and two sides, preferably bottom surface and side have at least 60 dynes per centimeter at least, the surface tension of at least 73 dynes per centimeter, at least 75 dynes per centimeter, at least 80 dynes per centimeter for example, perhaps for example the contact angle of water/target molecule less than 90 degree.

In an embodiment of microfluidic device, its interrupt flow channel section disposes to small part and is arrheaed by the capillary of its geometry setting, along the passage smallest cross-sectional size of cutout channel section greater than along at least one the smallest cross-sectional size of passage in contiguous branching channel section and the crystallization channel section.

In an embodiment of microfluidic device, its interrupt flow channel section disposes to small part and is arrheaed by the capillary of its geometry setting, the big area of section of area of section of at least one near this cutout channel section has in nearly branching channel section and the crystallization channel section.

Should be provided with how much arrange arrhea, make when filling the crystallization channel section because the speed of aforesaid liquid, mobilely can not stop.Yet when filling the crystallization channel section, liquid is because arrheaing of arranging for how much and can be prevented from flowing back into the cutout channel section.Those skilled in the art can provide arrheas, for example by selecting the proper angle at edge between the part of fluid passage.

In an embodiment of microfluidic device, fluid passage comprises the unexpected variation at least one cross section in the cutout channel section with contiguous branching channel section and crystallization channel section one or both next-door neighbours.

As well-known to those skilled in the art, such sharp edge may provide the capillary potential barrier, and it may postpone or even stop liquid to flow through this edge.

In an embodiment of microfluidic device, at least one cutout channel section has the big area of section of area of section of neighbour nearly crystallization channel section.The wall that fluid passage comprises around passage and passage length extends.In this embodiment, expectation by at least along the parameatal major part in the cutout channel section of next-door neighbour's crystallization channel section progressively the displacement wall form the larger cross-sectional area of at least one cutout channel section.

In an embodiment of microfluidic device, wherein fluid passage has polygon (quadrilateral polygonal) cross sectional shape on four limits, the polygonal cross-section shape on this four limit provides bottom surface, relative end face and two sides, compare with contiguous crystallization channel section, the larger cross-sectional area of at least one cutout channel section is to form by the bottom surface of the channel section that stops at least and the progressively displacement of side.

The progressively displacement of the described wall of the parameatal major part in preferred contiguous cutout channel section of edge at least and the crystallization channel section can preferably form step displacement (step displacement).

In one embodiment, preferred formation along the progressively displacement of the described wall of the parameatal major part in the cutout channel section of next-door neighbour's crystallization channel section at least has 120 degree or littler, for example the edge at 100 degree or littler, 90 degree or littler, 80 degree or littler angle.

The corner is measured as from the surface, limit of crystallization passage to the surface, limit on the rank that form progressively displacement, the minimum angle of whole solid structure.

The larger cross-sectional area of at least one cutout channel section is caused by the progressively displacement of the described wall of the parameatal major part in the preferred cutout of the edge at least channel section, and this larger cross-sectional area can have the arbitrary dimension greater than contiguous crystallization channel section area of section.The area of section that the area of section of at least one cutout channel section can preferably have a neighbour nearly crystallization channel section greatly at least 10%, for example at least 30%, at least 50%, at least 100%, at least 150%, at least 200%, 500%, at least 1000% size.

At the area of section of at least one cutout channel section in the area of section embodiment of contiguous crystallization channel section, for example 500% or bigger, the necessary amounts of the target molecule solution that is used to test will be big relatively, so that fill the crystallization channel section effectively.

In the only big slightly embodiment of the area of section of at least one cutout channel section, target molecule solution may increase from the risk that the crystallization channel section refluxes.Those skilled in the art can come the preferred size ratio of selector channel part with regard to concrete test type.In fact, best channel size ratio can depend on the kind and the availability of target molecule especially, and the character of target molecule solution, comprises surface tension.

In an embodiment of microfluidic device, fluid passage comprises the cutout channel section that at least one is provided by the passage displacement.Preferably, the cutout channel section is moved to than the higher position of adjacent at least crystallization channel section, and more preferably, at least one cutout channel section also is moved to the position higher than branching channel section.Described higher position is for the end face of device.In this embodiment, expect that at least one cutout channel section is moved to a level with respect to the crystallization channel section, when device remains on horizontal plane, prevents that liquid (for example water or target molecule sample) is back to the branching section from crystallized region thus.

In this embodiment, can expect except that by the displacement that the cutout channel section also has the neighbour nearly big area of section of crystallization channel section, and/or its how much ground form, and/or its surface nature provides less capillary force to water and/or target molecule solution.

In an embodiment of microfluidic device, wherein fluid passage comprises by the passage displacement and/or by setting having at least one cutout channel section that the cutout passage less than the capillary force of crystallization passage provides, this microfluidic device disposes floss hole, its during loading target molecule solution by for example stopper or bore closure, and after being filled, opens in the crystallization passage, to discharge excessive target molecule solution, provide or participate in providing the cutout in the cutout passage simultaneously.

In an embodiment of microfluidic device, solid structure disposes the line of weakness of intersecting with the fluid passage of at least one cutout in the channel section.This line of weakness can preferably be traversed whole basically device, for example traverses all basically end faces and/or bottom surface.This line of weakness intersect simultaneously two or more the cutout channel sections, for example all cutout channel sections.

Line of weakness is the line that has than intensity low around the solid structure, makes when device applies bending force, and this device can disconnect along described line of weakness.When device when line of weakness disconnects, this device also can break at the cutout channel section by described line of weakness intersection, and fluid passage can dispose along the fracture of microfluidic device one side thus.Because the small size of fluid passage, the liquid that can not in the fluid passage flows out like this, but owing to capillary force can be retained in the passage.For fear of evaporation of liquid, the mouth of passage can seal with sealant for example as described below.

In principle, the slit can have arbitrary form.For example, it can be the continuous slit that does not disconnect on length, and perhaps it can be a point-like.In one embodiment, line of weakness is that the form with the slit intersects at whole device at least one of end face and bottom surface, the slit preferred shape is in V-arrangement and the U-shaped, wherein said V/U shape can be about vertically axisymmetric in one embodiment, and in another embodiment can be not symmetrical like this.

The slit should be enough dark in the reduction of expectation to be provided, and makes device can disconnect along described slit when being subjected to bending force.In one embodiment, the degree of depth in slit is at least 10% of a microfluidic device thickness, for example its degree of depth be with the contiguous microfluidic device thickness in slit at least 15%, 30-90%, 35-80%, wherein the thickness of microfluidic device is defined as the distance between the relative bottom surface with it of end face.

Target molecule solution can directly join the inlet of target molecule solution, for example add by the inlet contact sample that makes target molecule solution, described sample for example places servicing unit or places on the servicing unit, for example wellhole or test tube, and perhaps the instrument by for example pipette applies.In one embodiment, microfluidic device of the present invention comprises the target molecule solution inlet cavity that is used for target molecule solution is fed into target molecule solution inlet.This target molecule solution inlet cavity can for example be configured as the depression of the solid structure end face that directly is directed at target molecule solution inlet.

In one embodiment, target molecule solution inlet cavity is the form of the cavity in the described end face, described cavity is by described target molecule solution inlet and described branching channel section fluid connection, and described target molecule solution inlet cavity preferably has the conical in shape of the inlet from end face to target molecule solution.

The volume that target molecule solution inlet cavity can have any desired.In one embodiment, the volume of target molecule solution inlet cavity is crystallization channel section cumulative volume at least, for example be at least 2 times, at least 1.5 times, at least 1.3 times, at least 1.2 times, at least 1.1 times of crystallization channel section volume, for example 0.05-20 μ l, 0.1-10 μ l, 0.2-5 μ l, 0.5-3 μ l.

In an embodiment of microfluidic device, with two or more precipitating reagents inlet be provided with separately with precipitating reagent cavity fluid connection.Device preferably includes at least one precipitating reagent cavity of each precipitating reagent inlet, yet in one embodiment, two or more precipitating reagent inlets and identical precipitating reagent cavity fluid connection.

In one embodiment, the precipitating reagent cavity except the opening of the inlet from the precipitating reagent cavity to precipitating reagent, is surrounded by installing.This refers to semiclosed cavity.In this embodiment, expectation has only the opening of precipitating reagent cavity to enter the mouth by precipitating reagent.Alternatively, precipitating reagent can add by the film on the precipitating reagent cavity wall, perhaps can with before the precipitating reagent cavity is provided the precipitating reagent cavity be filled in advance in that the element of solid structure is fixed to one another.Precipitating reagent for example can be to be in the precipitating reagent cavity of pre-filling with dry status.

In the embodiment that comprises semiclosed precipitating reagent cavity, device should preferably include the setting that allows air to escape when target molecule solution adding apparatus, so that target molecule solution enters the crystallization channel section.Being provided with like this can be the form of pore in for example installing, and it leads near the channel section of crystallization separately that preferably is arranged on the precipitating reagent inlet, or is arranged on the pore in the precipitating reagent cavity.These pores can comprise obturating element, plate for example, and glass plate for example, if expectation and when expecting, it can be used to stop up pore.

The precipitating reagent cavity can for example be configured as the depression in the solid structure end face that directly leads to the precipitating reagent inlet.

In an embodiment of microfluidic device, the precipitating reagent cavity is the form of the inlet cavity in described end face, described cavity is by described precipitating reagent inlet and described crystallization channel section fluid connection, and described precipitating reagent inlet cavity preferably has the conical in shape of the inlet from end face to target molecule solution.

In one embodiment, the minimum to each other distance of precipitating reagent cavity can be 0.1mm, for example 0.5-5mm, 1-3mm, 1.5-2.5mm at least.

In an embodiment of microfluidic device, at least one crystallization channel section has the capillary plug that is arranged on or is close to the precipitating reagent inlet, and this capillary plug preferably progressively provides, and preferably sharply enlarges to the precipitating reagent cavity from the crystallization channel section.Preferably, all crystallization channel section has and is arranged on or contiguous their the capillary plug of precipitating reagent inlet.

Therefore, can target molecule solution be filled into the crystallization channel section from target molecule solution inlet by utilizing capillary force, and not fill the precipitating reagent cavity simultaneously.At the capillary plug place that is equipped with, the liquid front end of target molecule solution will stop to flow.If precipitating reagent is filled in the precipitating reagent cavity subsequently, target molecule solution and precipitating reagent will be in contact with one another diffusion so, and can carry out crystallization trial.

Can be preferably formed from having 120 degree or littler to the progressively expansion of precipitating reagent cavity from the crystallization channel section, for example the edge at 100 degree or littler, 90 degree or littler, 80 degree or littler angle.

The corner is measured as from the surface, limit of crystallization passage to the surface, limit on the rank that form progressively displacement, the minimum angle of whole solid structure.

The precipitating reagent cavity can have any desired volume, and for example volume is 0.01-10 μ l, for example 0.02-5 μ l, 0.5-2 μ l, 0.7-1.3 μ l.

Microfluidic device can be made by independent parts, but the microfluidic device of such parts is difficult to prepare usually.Preferably, microfluidic device is by being equal to or two or more parts of different materials are made, and its parts are connected to each other.These parts can be connected to each other in any way, for example bonding, welding and/or mechanical grip.

Connection can be for example to relate to the chemical bond that one or more are selected from the key of ionic bond and covalent bond.In one embodiment, the connection between two or more parts is finished by welding, and for example the hot pressing connection is connected with ultrasonic.

Microfluidic device can be made by the material of any appropriate.The various parts of microfluidic device can be made by that be equal to or different materials.In one embodiment, microfluidic device comprises at least one by the parts that the material that is selected from glass, pottery, metal, semi-conducting material and polymer is made, and described semi-conducting material is for example silicon, germanium and GaAs.

Semi-conducting material is the material (partly filling up) that its skin has 4 electronics.When being bonded together in lattice, the atom shared electron makes them have 8 electronics at skin separately.Electronics is the loosely bonding to a certain extent, so they can become carrier in the presence of electric field.

In one embodiment, microfluidic device comprises the parts that at least one is made by semi-conducting material, and this semi-conducting material is selected from the single-crystal silicon Germanium (SG) of the polysilicon of doping, the polycrystalline silicon germanium of doping (poly SG), the monocrystalline silicon that mixes, doping and the III-V family material that mixes.

In one embodiment, microfluidic device comprises the parts that at least one is made by polymeric material, be preferably the polymer of injection moldable, for example be selected from following polymer: acrylonitrile-butadiene-styrene copolymer, Merlon, dimethyl silicone polymer (PDMS), polyethylene (PE), polymethyl methacrylate (PMMA), polymethylpentene, polypropylene, polystyrene, polysulfones, polytetrafluoroethylene (PTFE), polyurethanes (PU), polyvinyl chloride (PVC), poly-inclined to one side 1,1-dichloroethylene (PVDC), poly-inclined to one side 1, the 1-difluoroethylene, styrene-propene acid copolymer (styrene-acrylcopolymers), polyisoprene, polybutadiene, polychlorobutadiene, polyisobutene, poly-(s-B-S), siloxanes, epoxy resin, polyether block amide, polyester, acronitrile-butadiene-styrene (ABS), acrylic acid, celluloid, cellulose acetate, ethane-acetic acid ethyenyl ester (EVA), ethylene-vinyl alcohol (EVAL), fluoroplastics (PTFEs, comprise FEP, PFA, CTFE, ECTFE, ETFE), polyacetals (POM), polyacrylate (acrylic acid), polyacrylonitrile (PAN), polyamide (PA), polyamide-imides (PAI), PAEK (PAEK), polybutadiene (PBD), polybutene (PB), polybutylene terephthalate (PBT), PET (PET), poly terephthalic acid hexamethylene dimethyl ester (PCT), polyketone (PK), polyester/polythene/polyethene, polyether-ether-ketone (PEEK), PEI (PEI), polyether sulfone (PES), polychlorostyrene ethene (polyethylenechlorinates) (PEC), polyimides (PI), PLA (PLA), polymethylpentene (PMP), polyphenylene oxide (PPO), polyphenylene sulfide (PPS), polyphthalamide (PPA) and composition thereof, preferably Zhuan Zhi at least one parts are transparent, are used for optical check.

When using polymer, in one embodiment, the Young's modulus E of preferred polymers is 0.2GPa at least, for example 0.5GPa, 1GPa at least at least.Therefore material can have the reliable hardness that is used to carry out crystallization.

In one embodiment, microfluidic device comprises the parts that at least one is made by cyclic olefine copolymer, for example PET,

(selling) by ZEON company,

(selling) by ZEON company and

(selling) by Topas Advanced Polymers GmbH.These materials are particularly useful for the production of removable hood as described below.

In one embodiment, wherein microfluidic device is made by at least two parts connected to one another, and at least one in these parts comprises the line of weakness of slit form.

In one embodiment of the invention, microfluidic device is made by bottom parts (bottom surface is provided) and top part (end face is provided) at least, and at least one in bottom parts and the top part comprises the groove that fluid passage is provided.Groove should be arranged on respectively with bottom surface and end face facing surfaces on.In addition, at least one in bottom parts and the top part comprises the hole of target molecule solution inlet cavity and precipitating reagent inlet cavity, and described bottom parts and described top part are preferably rigidity substantially.

In one embodiment, thus microfluidic device by making by bonding, welding and/or mechanical grip bottom parts and the top part that forms the fluid passage between parts fixed to one another.

Usually, it is the simplest two parts being welded together.Yet in an embodiment that wherein two parts is clamped together, the following open crystal of collecting growth may be very simple.

In one embodiment, microfluidic device is made by the seal member of bottom parts, top part and sealant form at least.Top part comprise be used to provide fluid passage in the groove of the relative side of end face, the randomly hole of target molecule solution inlet cavity and the hole/depression of precipitating reagent cavity.Seal member is sandwiched between top part and the bottom parts, makes seal member cover described groove and described hole, and the described seal member of base supports.In this embodiment, bottom parts can be preferably the form of common substantially plate, is preferably made by glass or polymer.

The groove that the preferably described at least therein sealant of bottom parts covers top part supports sealant with the zone that forms the crystallization channel section.Described top part can comprise the line of weakness of one or more transcrystallization channel sections, and described line of weakness can be preferably the part of the top part that extends bottom parts, makes device described line of weakness to disconnect in the situation lower edge that bottom does not disconnect.

In one embodiment, wherein microfluidic device is made by the seal member of bottom parts and sealant form at least, and top part comprises and is used to provide the groove of fluid passage and the randomly hole of target molecule solution inlet cavity and the hole/depression of precipitating reagent cavity.Seal member is connected with top part makes seal member cover described groove and described hole.Thereby can form fluid passage.

Should select seal member to be made by the material that necessary tight ness rating can be provided, or alternatively be made by sealant, for example wax can be used for sealing device.In one embodiment, seal member is made by polymeric material, and the Shore A hardness of this polymeric material is 90 or lower, and for example Shore A hardness is 20-70.

Example as the preferred polymers material that is used for seal member can be mentioned elastomer, more preferably is selected from the elastomer of thermoplastic elastomer (TPE) and rubber.The material that is suitable for seal member is PDMS.

In one embodiment, seal member can be preferably the form of common substantially plate.

In one embodiment, microfluidic device comprises also and preferably intersects the line of weakness of all crystallization channel sections that device can disconnect along described line of weakness thus.Line of weakness can be by above-mentioned outfit, and for example this line of weakness can be provided in the top part of microfluidic device.

Line of weakness can preferably be arranged in the described structure, closes on the described crystallization channel section of the described inlet of precipitating reagent separately with intersection.

For the simple production and the operation of microfluidic device, in one embodiment, end face and opposed bottom surface can be smooth substantially, except the inlet cavity with randomly precipitating reagent cavity and/or the line of weakness.

The capillary performance of device depends on the some factors as being well known to those skilled in the art.As mentioned above, capillarity depends on the structure and the size of surface tension, fluid passage.Term " capillary dimensions " is meant that fluid passage has such character, and it can cause the capillary pulling power in the such fluid sample of water and/or target molecule solution.

In an embodiment of microfluidic device, fluid passage has capillary dimensions on the major part of its length, and preferably, at branching channel section and crystallization channel section at least, described fluid passage has capillary dimensions.

In an embodiment of microfluidic device, fluid passage has at least one sectional dimension less than 1000 μ m on the major part of its length, for example 10-250 μ m, 20-100 μ m.Preferably, at branching channel section and crystallization channel section at least, described fluid passage has at least one sectional dimension less than 1000 μ m, for example 10-250 μ m, 20-100 μ m.

In an embodiment of microfluidic device, fluid passage is at least at described crystallization channel section, polygon (quadrilateral polygonal) cross sectional shape that preferably on its whole length, has four limits that bottom surface, relative end face and two sides are provided, the polygonal cross-section shape on this four limit is preferably trapezoidal, described two sides have the angle between the 90-120 degree of bottom surface, for example 90-110 degree, 90-100 degree, 90-95 degree.

In an embodiment of microfluidic device, fluid passage has the cross sectional shape of width and height at least at described crystallization channel section, highly be the average distance between bottom surface and end face, and width is the average distance between two sides, this width and be preferably 10-300 μ m highly separately from each other.

The branching channel section can comprise one, two, the site of three or more branches, and on the site of each branch, passage can be branched off into two or more a plurality of passage.

The type of the cutout that provides is provided the optimum length of cutout channel section to a great extent.Usually, the length of expectation cutout channel section is 1-10000 μ m, for example 5-5000 μ m, 10-1000 μ m, 20-200 μ m.

In an embodiment of microfluidic device, wherein said cutout setting is the form of the line of weakness of intersection fluid passage, the length of described cutout channel section is up to about 5000 μ m, for example be up to about 4000 μ m, be up to about 3000 μ m, be up to about 1000 μ m, be up to about 1000 μ m, be up to about 100 μ m, for example 1-50 μ m, for example at least 10 μ m.This length is defined as the length of the fluid passage that is intersected by line of weakness, wherein with end face abreast (in planewith the top face) measure the width of line of weakness.

In an embodiment of microfluidic device, wherein said cutout setting is the form of the capillary plug in the fluid passage, and the length of described cutout channel section is up to about 10mm, for example 1-10000 μ m, 10-5000 μ m, 20-2000 μ m.

The crystallization channel section can preferably have the length of 10-50000 μ m separately, for example 20-30000 μ m, 40-15000 μ m, 100-10000 μ m.

For simplified structure, described crystallization channel section can preferably be set parallel to each other substantially.

In one embodiment of the invention, microfluidic device comprises that first parts that have first at least are used to provide the groove face of the groove of the crystallization channel section of fluid passage at least with having, and at least one provides many covers to the small part removable hood, have second and groove face separately, wherein described removable hood is applied on the groove and along the groove sealing so that the crystallization channel section of fluid passage at least to be provided.

In one embodiment, first of first parts is end face, and one second of cover or a plurality of second face provide the bottom surface.In another embodiment, first of first parts is the bottom surface, and one second of cover or a plurality of second face provide end face.

First parts can be the object of for example molded and/or micromachined.In one embodiment, first parts are made by flat substantially plate, and this flat board has been used laser grooving.By utilizing laser, this groove can be shaped to have the shape of any desired in practice in very simple mode.

One or more covers are applied on the groove to form fluid passage.In fact, be desirably in and apply shutter fitting in one and during it applies or afterwards provide the cutting of expectation so that movably lid to be provided subsequently to cover.Movably lid is a kind of like this lid, and it can move to expose at least a portion crystallization channel section by tearing, preferably exposes the major part of at least one crystallization channel section.

By this movably lid, the big molecule of crystallization can be collected in very simple mode.In addition, the lid that only covers the macromolecular crystallization channel section of crystallization with expectation need be exposed collecting, and remaining crystallization channel part can allow the further crystallization of extra time.In other words, the crystal of expectation is just collected in the further crystallization that does not need not have by the time to expect.

Target molecule solution inlet can be equipped with as mentioned above.Preferably, target molecule solution inlet is provided by a through hole in first parts.Alternatively, target molecule solution inlet is provided by above-mentioned depression.

The precipitating reagent inlet also can be aforesaid.In one embodiment, first parts have at least one provides through hole from the precipitating reagent inlet to fluid passage.Alternatively, the precipitating reagent inlet is provided by above-mentioned depression.

By the inlet of through-hole form is provided in first parts, microfluidic device can provide in very simple mode.

Movably lid can preferably extend first parts, for example makes removable hood be provided for moving separately the flange of lid separately.

In one embodiment, removable hood comprises the flange that is not adhered on first parts separately, and this flange is set makes to move separately lid from crystallization channel section separately by tearing in this flange.Like this, the crystallization channel section that can expose expectation is collected in mode simply and easily.

Movably lid can preferably pass through the groove face of bonding and/or welded seal to the first parts along groove separately.Potted line can be the line of for example relative thin, for example less than 0.5mm, and 2mm or littler for example, it for example provides the part or all of cutting between the contiguous removable hood simultaneously.The potted line of broad also can be used naturally, but in this case, the independent step of cutting between contiguous removable hood may be necessary, and perhaps single movably lid can be used separately.

In one embodiment, movably lid for example releasably is connected to each other along potted line.Should preferably have the low in fact intensity of inherent strength along the thrown off connection of potted line between contiguous removable hood than the material of making this lid, for example be make this lid material inherent strength half or be lower than the inherent strength of the material of making this lid, the connection meeting that this can be thrown off when making tearing disconnected before the tear-off lid material.

In one embodiment, microfluidic device comprises 3 or more removable hoods.The number of removable hood can preferably be adjusted with the number of crystallization channel section.Preferably, microfluidic device comprises the removable hood that is used for each described crystallization channel section.

Microfluidic device for example can be provided by any above-mentioned material.In one embodiment, the material of making first parts is selected from glass, pottery, metal, semi-conducting material and polymer, described semi-conducting material is silicon, germanium and GaAs for example, and described polymer has the Young's modulus of 0.2GPa at least, for example 0.5GPa, 1GPa at least at least.First parts preferably can be for transparent substantially.

One or more covers can preferably be made by paper tinsel, for example polymer foil or metal forming.The expectation material of cover is more than disclosed.

In one embodiment, the paper tinsel that will be used to cover is applied to the groove face of first parts, and it is bonding or be welded on the groove face along groove, and it partially or even wholly is cut into pieces so that described removable hood to be provided.Other method that is used to produce also can be used naturally and be available to those skilled in the art.

In one embodiment, microfluidic device also comprises the film of the described crystallization channel section of introducing at least one preferably contiguous described inlet of precipitating reagent separately.

Film for example can reduce the amount of moving out and entering the target molecule of precipitating reagent by the precipitating reagent inlet from the crystallization channel section.In many cases, target molecule is greater than precipitating reagent, and its meaning is that precipitating reagent can pass through film, but target molecule can not.

Film for example can be to prevent that target molecule is diffused into the filter membrane in the precipitating reagent that contacts with precipitating reagent inlet liquid, described film preferably can be the perforated membrane of being made by one or more materials, this material is selected from fibrous material, for example melt jet glass fibre, spunbond synthetic fibers (for example, nylon, polyester, polyethylene, polypropylene or polyether sulfone), semisynthetic fibre, regenerated fiber and inorfil and mixture; And foamable polymer, more preferably described film is a porous celluloid.

In one embodiment, film can have surface tension, and this surface tension more approaches the surface tension of precipitating reagent than target molecule solution.

In one embodiment, microfluidic device and target molecule solution combination.This target molecule solution for example can be for as disclosed herein, for example the aqueous solution.This aqueous solution can not contain washing agent or randomly can comprise one or more washing agent.

Can preferably provide target molecule solution to microfluidic device, the meaning is that the capillary force of the expectation in the fluid passage should be with respect to the target molecule measured in solution.Therefore, in one embodiment, microfluidic device comprises at least one cutout channel section, this cutout channel section is set provides capillary force with the target molecule solution in described passage, and this capillary force is less than the capillary force that is provided to target molecule solution by described contiguous crystallization channel section.

In the embodiment of a microfluidic device and the combination of target molecule solution, the wall that described fluid passage comprises around passage and passage length extends.In at least one cutout channel section, in part around it at least, the surface tension of described wall is lower than the surface tension of target molecule solution.At this or in another embodiment, at least one crystallization channel section, in part around it at least, the surface tension that described wall can have is higher than the surface tension of target molecule solution.

In the embodiment of a microfluidic device and the combination of target molecule solution, the wall that described fluid passage comprises around passage and passage length extends.Described fluid passage has the polygonal cross-section shape on four limits that bottom surface, relative end face and two sides are provided at least in a crystallization channel section, preferably the surface tension of bottom surface and side is higher than the surface tension of target molecule solution at least.

In the embodiment of a microfluidic device and the combination of target molecule solution, the wall that described fluid passage comprises around passage and passage length extends.At described branching channel section, in part around it at least, the surface tension of described wall is higher than the surface tension of target molecule solution.

In the embodiment of a microfluidic device and the combination of target molecule solution, fluid passage has capillary dimensions in the major part of its length, preferably described fluid passage has capillary dimensions at branching channel section and crystallization channel section at least, and this capillary dimensions is definite with respect to target molecule solution.

Below further disclose the microfluidic device with target molecule solution combination, any relation between flowing liquid of wherein being fit in fluid passage and the passage is with respect to the target molecule measured in solution.

The invention still further relates to a kind of method of utilizing above-mentioned microfluidic device to promote target molecule crystallization from target molecule solution.

Method of the present invention comprises:

I) provide target molecule solution, with the target molecule solution feed to target molecule solution inlet and make it fill up the crystallization channel section;

At least a precipitating reagent ii) is provided and is provided with to make itself and at least one precipitating reagent fluid connection that enters the mouth, and it is contacted with target molecule solution;

Iii) the crystallization channel section make precipitating reagent diffuse into target molecule solution and

Iv) the fluid connection between the described crystallization of target molecule separately channel section is disconnected.

Step I) and step I i) can carry out with random order.

Step I v) can be in step I) any time after having stopped carries out.

In principle, target molecule can be the molecule of the expectation crystallization of any kind.Target molecule can be inorganic or organic.Described molecule is generally biomolecule most, promptly is derived from the molecule of biological sample or its artificial analog.Preferably, target molecule solution is the solution of at least a target molecule, and this target molecule is selected from protein, nucleic acid, nucleic acid analog, carbohydrate, lipid, more preferably be selected from 500 dalton or more protein, strand and double-stranded DNA, RNA, PNA and LNA, and drug candidate.Term protein comprises peptide and bigger protein.

In preferred embodiments, target molecule is for being selected from 500 dalton or more protein.

As mentioned above, solution can comprise other composition, for example is used for the composition of stabilizing solution, for example as the polymer of polyethylene glycol, and the surfactant that comprises washing agent.

The concentration of various compositions can change widely.

The example of washing agent and concentration sees Table 1.Usually, target molecule solution can only comprise one type washing agent; Yet the combination of washing agent also can be used.

MW: molecular weight

CMC: critical micelle concentration

Actual: used general concentration

Table 1:

In one embodiment, gelling agent can be joined in the target molecule solution, make partly solidified at least the backflow of this solution to prevent from not expect.The example of useful gelling agent has agarose and acrylamide.

Being combined as of useful precipitating reagent and precipitating reagent is well known in the art.As described above, precipitating reagent can be used with the form of drying or the form of solution.

The example of precipitant solution is found in Shotgun crystallization strategy for structuralgenomics:an optimized two-tiered crystallization screen against the Thermotogamaritima proteome ', Page R, Grzechnik SK, Canaves JM, Spraggon G, KreuschA, KuhnP, Stevens RC, Lesley SA work, ACTA CRYSTALLOGRAPHICASECTION D-BIOLOGICAL CRYSTALLOGRAPHY 59:1028-1037, the 6th part, in June, 2003.

Those skilled in the art will understand and how find and select and precipitating reagent that the target molecule solution selected is used in combination.

In one embodiment, target molecule solution can directly be fed into target molecule solution inlet, for example utilizes instrument, and pipette for example is perhaps by putting on target molecule solution in the aforesaid target molecule solution inlet cavity.

In one embodiment, target molecule solution for example puts on support with the form of drop, target molecule solution inlet is configured as the inlet that leads to capillary channel, and by target molecule solution inlet is contacted with drop, this drop can directly be drawn onto in the microfluidic device by capillary force.

For target molecule solution safe, not contaminated and that measure is provided, preferred microfluidic device comprises target molecule solution cavity, and target molecule solution is put on described target molecule solution cavity.

Because the structure/surface characteristic of microfluidic device fluid passage, target molecule solution can fill up the crystallization channel section.

In one embodiment, precipitating reagent is pre-charged with in the precipitating reagent cavity.Disclosed as mentioned, in this embodiment, precipitating reagent can be preferably drying regime.Precipitating reagent can be dissolved before or after inserting target molecule solution again, for example by injecting fluid in the precipitating reagent cavity, for example passed through film.Other example is more than disclosed.

In one embodiment, be provided with and make precipitating reagent and separately precipitating reagent inlet fluid connection in precipitating reagent is applied to contain precipitating reagent with the precipitating reagent inlet by will be separately the hole.

In one embodiment, precipitating reagent is set to make precipitating reagent and the inlet of precipitating reagent separately fluid connection by the precipitating reagent cavity that precipitating reagent is fed in the device.In this embodiment, precipitating reagent inlet can preferably be set makes target molecule solution flow into the crystallization channel section but does not flow into the precipitating reagent cavity.Such particular instance has more than been described.In this embodiment, target molecule solution applied before precipitating reagent.

In another embodiment, for example prevent that by above-mentioned film or capillary plug precipitating reagent from flowing into the crystallization channel section.In this example, precipitating reagent can preferably apply before target molecule solution.

According to this method, microfluidic device can be preferably like this, promptly because the displacement of the interior capillary force of aforesaid fluid passage or cutout channel section, and the fluid connection automatic disconnection between the described crystallization of target molecule separately channel section.

In one embodiment, disconnect the fluid connection disconnection that makes between the described crystallization of target molecule separately channel section by installing along the line of weakness of intersection cutout channel section.When device when line of weakness disconnects, the liquid in the crystallization channel section is owing to the capillary force in the crystallization channel section will can not flow out.

In one embodiment, allow precipitating reagent to diffuse into target molecule crystallization channel section, keep the time of expectation, for example 1 hour or longer, after this, the fluid connection between precipitating reagent and target molecule crystallization channel section stopped.

In order to prevent that liquid from evaporating from system, expectation will be led to the channel seal of microfluidic device surrounding environment.Therefore, this method preferably includes the inlet of at least one crystallization channel section of sealing, and the inlet of all crystallization channel sections of preferably sealed evaporates thus avoiding.

Can use any potted component/material.In one embodiment, one or more inlets are sealed by one or more following modes

A) by adding wax, for example paraffin or Tissuemat E come sealed entry; With

B) by fixing (for example, by bonding, welding or clamp) blocking element, the blocking element of making by polymer or glass for example, for example polymer sheet or sheet glass, this blocking element is preferably transparent.

This method also can preferably include cultivates microfluidic device and crystal is formed and/or growth.The cultivation time is depended on the type of target molecule solution.Generally, the most common cultivation time will be 2-580 hour, for example 24-240 hour.Cultivate and generally carry out in temperature-controlled cabinet, for example temperature is 25 degree, 16 degree or 4 degree.Temperature may influence crystallization, and tests for some, can carry out the cultivation under the different temperatures.

After cultivation, microfluidic device is checked,, differentiated that any crystal forms for example visually or by robot.The crystal that forms can be checked in fluid passage, for example by transparent wall part, perhaps can collect them and do further check.

In one embodiment, method of the present invention also comprise from one or more crystallization channel sections collect form/crystal that grows up to, preferably, crystal can be collected by following one or more modes:

A) by at least two parts in the disconnecting apparatus separately forming the parts of crystallization channel section, and shift out crystal;

B), and shift out crystal by selectively shifting out (for example, cutting) one or more bottom parts or top parts of selecting the crystallization channel section;

C) by from the crystallization channel section, drawing or extrude crystal; With

D) by shifting out removable hood the crystal channel section of expectation is exposed, thereby can shift out crystal.

Aspect second, the present invention relates to be used to promote the another kind of microfluidic device of target molecule crystallization.This microfluidic device comprise have first with second relative and the solid structure of at least one fluid passage, wherein said fluid passage comprises target molecule solution inlet and at least one precipitating reagent inlet.In other words, in this microfluidic device, do not need branching section and/or cutout section.This target molecule solution inlet is by described fluid passage and precipitating reagent inlet fluid connection, and fluid passage comprises the crystallization channel section of contiguous described precipitating reagent inlet.This microfluidic device of second aspect present invention comprises that at least one has first first parts, has the groove face of at least one groove that is used to provide described crystallization channel section and at least one provides to the cover of small part removable hood, described lid has second and groove face, and wherein said removable hood is applied on the described groove and along described groove and seals so that the crystallization channel section of described fluid passage to be provided.

In one embodiment, first face of first parts is end faces, and one second of cover or a plurality of second face provide the bottom surface.In another embodiment, first face of first parts is bottom surfaces, and one second of cover or a plurality of second face provide end face.

At least one is transparent in first parts or the cover.

First parts of this second aspect of the present invention can be made by above disclosed material, and can be provided by similar methods.First parts can be the object of for example molded and/or micromachined.In one embodiment, first parts are made by flat board substantially, and this flat board is by laser grooving.By utilizing laser, can form groove and make it have the shape of any desired in practice in very simple mode.

Cover is applied on the groove to form fluid passage.In practice, be desirably in and apply cover in one and subsequently during it applies or afterwards for providing one or more movably lids to provide any desired cutting to cover.Movably lid is a kind of like this lid, and it can be moved by tearing and expose at least a portion crystallization channel section, preferably exposes the major part of at least one crystallization channel section.

By this movably lid, the big molecule of crystallization can be collected in very simple mode.In addition, only cover lid and need be exposed and collect, and remaining crystallization channel section can allow the further crystallization of extra time with the macromolecular crystallization channel section of expectation crystallization.In other words, the crystal of expectation is just collected in the further crystallization that does not need not have by the time to expect.

Target molecule solution inlet can be equipped with as mentioned above.Preferably, target molecule solution inlet is provided by the through hole in one first parts, and alternatively, target molecule solution inlet is provided by aforesaid depression.

The precipitating reagent inlet also can be aforesaid.In one embodiment, first parts have at least one provides through hole from the precipitating reagent inlet to fluid passage.Alternatively, the precipitating reagent inlet is provided by aforesaid depression.

By one or more inlets of through-hole form are provided in first parts, microfluidic device can provide in very simple mode.

Movably lid can preferably extend first parts, for example makes one or more movably lids be provided for moving separately the flange of lid separately.

In an embodiment of second aspect, described movably lid comprises the flange that is not adhered on first parts, and this flange is set makes and to draw the flange place to move described lid from the crystallization channel section by this.So, can expose desired crystallization channel section and collect in mode simply and easily.

Movably lid can be preferably along separately groove by bonding and/or welded seal groove face to first parts.Potted line can be the pipeline of for example relative thin, for example less than 0.5mm, and 2mm or littler for example, it for example can provide the part or all of cutting between the optional contiguous removable hood simultaneously.The potted line of broad also can use naturally.

In one embodiment, the microfluidic device of second aspect comprises two or more fluid passages, and this fluid passage comprises the crystallization channel section and is connected with the precipitating reagent inlet with target molecule solution inlet separately.Preferably by movably lid covering, this lid randomly releasably is connected with each other each described crystallization channel section.

Under the situation of two or more removable hoods, this lid can be aforesaid.In one embodiment, microfluidic device comprises two or more, for example 5 or more a plurality of, and 10 or more a plurality of crystallization channel section, and preferably, described microfluidic device comprises the lid that is used for each described crystallization channel section.

The microfluidic device of second aspect for example can be provided by above-mentioned any material.In one embodiment, first parts are made by the material that is selected from glass, pottery, metal, semi-conducting material and polymer, described semi-conducting material is silicon, germanium and GaAs for example, and described polymer has the Young's modulus of 0.2GPa at least, for example 0.5GPa, 1.0GPa at least at least.First parts preferably can be for transparent substantially.

At least one cover is made by paper tinsel, for example polymer foil or metal forming.The expectation material that is used for cover is more than disclosed.

In an embodiment of second aspect, the paper tinsel that will be used to cover puts on the groove face of first parts, and is bonding or be welded on the groove face along groove, and partially or even wholly is cut into pieces so that described removable hood to be provided.Other production method also can be used naturally and be available for those skilled in the art.

The microfluidic device of second aspect in addition can be combined with above disclosed any parts, as long as it comprises at least one aforesaid removable hood.

The accompanying drawing summary

Below, will embodiment of the present invention be described more fully with reference to accompanying drawing, wherein:

Fig. 1 a represents according to microfluidic device schematic diagram of the present invention.

Fig. 1 b and Fig. 1 c represent the to stop example of channel section.

Fig. 2 a is the top view of microfluidic device of the present invention, and this microfluidic device is included in the cutout channel section of the line of weakness form between branching channel section and the crystallization channel section.

Fig. 2 b is the side sectional view of the microfluidic device shown in Fig. 2 a along A-A ' cutting line.

Fig. 3 a is the top view of the another kind of microfluidic device of the present invention, and this device is included in the cutout channel section of the line of weakness form between branching channel section and the crystallization channel section and the cutout channel section of other line of weakness form in the precipitating reagent porch.

Fig. 3 b is the side sectional view of the microfluidic device shown in Fig. 3 a along A-A ' cutting line.

Fig. 4 a-d is the side sectional view of the various states of another kind of microfluidic device of the present invention in its use.

Fig. 5 a-b is the side sectional view of the various states of a part in its use of another kind of microfluidic device of the present invention.

Fig. 6 a-b is the top part of microfluidic device of the present invention.

Fig. 7 a-b is respectively first parts of microfluidic device of the present invention and complete microfluidic device.

Fig. 8 is the schematic diagram of microfluidic device of the present invention.

Fig. 9 represents the microfluidic device of second aspect present invention.

Figure 10 represents the top view of the another kind of microfluidic device of the present invention.

For the sake of clarity, figure is schematically and is simplified, and they just show understanding the details of necessity of the present invention.And omitted other details.In the text, identical reference number is used for identical or corresponding parts.

Fig. 1 has schematically represented the present invention, and its expression microfluidic device 1 comprises fluid passage 2.This fluid passage 2 comprises inlet 3 and target molecule solution inlet 4, and this target molecule solution inlet 4 randomly has target molecule solution inlet cavity.Described fluid passage 2 also comprises precipitating reagent inlet 5, and this precipitating reagent inlet 5 randomly has precipitating reagent cavity 6, and this precipitating reagent cavity 6 randomly covers with the lid that does not illustrate, and described lid for example is aforesaid form membrane.

Fluid passage 2 comprises three parts, i.e. branching channel section 7, cutout channel section 8 and crystallization channel section 9.In Fig. 1, show only two crystallization channel sections, but as mentioned above, this microfluidic device can have many as expected crystallization channel sections.

In use, target molecule solution is fed to target molecule solution inlet 3 by the inlet of target molecule solution for example cavity 4, and target molecule solution is with flow channel 2 and be full of crystallization channel section 9.Before or after the target molecule solution feed is gone into fluid passage 2, precipitating reagent is fed into precipitating reagent cavity 6.If target molecule solution was fed into system before precipitating reagent, provide aforesaid setting to flow into the precipitating reagent cavity so to prevent target molecule solution.If precipitating reagent was fed into system before target molecule solution, provide aforesaid setting to be full of the crystallization channel section so to prevent precipitating reagent, for example precipitating reagent can provide with drying regime.

When target molecule solution and precipitating reagent all are fed into system, they will enter the mouth 5 or near contact each other it at precipitating reagent, and precipitating reagent will be diffused in the target molecule solution, and it causes the target molecule crystallization the most at last when cultivating.

After target molecule solution had been filled crystallization channel section 9, the fluid connection between branching channel section 7 and crystallization channel section 9 disconnected, and makes various precipitating reagents can not flow into contiguous crystallization channel section 9.Cutout channel section 8 is set because its structure and/or surface characteristic oneself disconnect fluid connection by it, or provide the line of weakness that does not illustrate make the user in simple and safe mode along such cutout line disconnecting apparatus, thereby and disconnect fluid connection between branching channel section 7 and the crystallization channel section 9.

As mentioned above, any opening from crystallization channel section 9 to surrounding air is by adding the sealant sealing that does not illustrate.

Fig. 1 b and Fig. 1 c represent the to stop example of channel section 8a, 8b, it can be the figure along the A-A ' cutting line of Fig. 1.

Fig. 1 b is the cutaway view of cutout channel section 8a.Flow direction when arrow represents that target molecule solution enters the mouth 3 chargings by target molecule solution.Described cutout channel section 8a is the form with the passage displacement, and wherein passage is with respect to contiguous branching channel section 7 and 9 vertical displacements of crystallization channel section.When target molecule solution when branching channel section 7 flows out because the speed of liquid, this target molecule solution can be easily by cutout channel section 8a and enter crystallization channel section 9.When liquid stopped to flow, just in case the branching channel section does not fill up (this is not right) fully, all the other liquid among the channel section 8a that stops so will flow back into the branching channel section.Because the sharp-pointed limit ' a ' that this progressively displacement of wall forms (for example has 120 degree or littler angles, for example 100 degree or littler angle, 90 degree or littler angle, 80 are spent or littler angles), so target molecule solution will be retained in the crystallization channel section 9, but cutout channel section 8a will drain basically.

Fig. 1 c is the cutaway view of another example of cutout channel section 8b.Flow direction when arrow represents that target molecule solution enters the mouth 3 chargings by target molecule solution.Cutout channel section 8b is the form of wall displacement, this wall centers on and formation cutout channel section 8b, wherein increase the sectional dimension of resolving flow passage section 8b, the capillary pulling power among the feasible cutout channel section 8b is less than the contiguous branching channel section 7 and the capillary pulling power of crystallization channel section 9.When target molecule solution when branching channel section 7 flows out because the speed of liquid, this target molecule solution can be easily by cutout channel section 8b and enter crystallization channel section 9.When liquid stopped to flow, owing to the difference of capillary pulling power in stop separately channel section and branching channel section, all the other liquid among the cutout channel section 8b will flow back into the branching channel section.Because the sharp-pointed limit ' b ' that this progressively displacement of wall forms (for example has 120 degree or littler angles, for example 100 degree or littler angle, 90 degree or littler angle, 80 are spent or littler angles), so target molecule solution will be retained in the crystallization channel section 9, but cutout channel section 8b will drain basically.

Fig. 2 a and 2b represent microfluidic device of the present invention, and this device comprises the cutout channel section 28 with the line of weakness form between branching channel section 27 and the crystallization channel section 29.Shown in Fig. 2 b, this device is to provide with three layers of form that following three parts form:

● top part 21a, have end face 21a ' and comprise the groove that is used to provide fluid passage 22, be used for the hole and the solution inlet cavity 24 of the precipitating reagent cavity 26 of the contiguous inlet of precipitating reagent separately 25, and the slit 28a that is used for providing in cutout section 28 line of weakness.

● bottom parts 21c, with the form of common plate, for example glass plate substantially.

● seal member 21b, to be clipped in the form of the sealant between top part 21a and the bottom parts 21c, make seal member cover described groove 22,26,24,28 and hole, and the described seal member 21b of base supports.Bottom parts 21c supports seal member 21b, and wherein seal member covers the groove 22 of the first parts 21a in the channel parts that forms crystallization channel section 29, but bottom parts 21c does not extend the cutout channel section 28 that is formed by slit 28a.Therefore, microfluidic device is not needing to disconnect under the situation of bottom parts 21c and can easily disconnect along line of weakness.

Fig. 3 a and 3b represent the another kind of microfluidic device of the present invention.This microfluidic device is the variant of microfluidic device shown in Fig. 2 a and the 2b, and be included in the cutout channel section 38 of the line of weakness form between Zhi Zhihua channel section 37 and the crystallization channel section 39 and at the enter the mouth other cutout channel section 40 of line of weakness form at 35 places of precipitating reagent.This device is to provide with three layers of form that following three parts form:

● top part 31a, have end face 31a ' and comprise the groove that is used to provide fluid passage 32, be used for the hole and the solution inlet cavity 34 of the precipitating reagent cavity 36 of the contiguous inlet of precipitating reagent separately 35, and be used for the slit 38a of line of weakness is provided and is used for providing the slit 40a of line of weakness at precipitating reagent 35 places that enter the mouth in cutout section 38.

● bottom parts 31c, with the form of common plate, for example glass plate substantially.

● seal member 31b, to be clipped in the sealant form between top part 31a and the bottom parts 31c, make seal member cover described groove 32,36,34,38,40 and hole, and the described seal member 31b of base supports.Bottom parts 31c supports seal member 31b, wherein seal member covers the groove 32 of the first parts 31a in the channel parts that forms crystallization channel section 39,, but bottom parts 31c does not extend cutout channel section 38 that is formed by slit 38a and the cutout section 40 that is formed by slit 40a.Therefore, microfluidic device is not needing to disconnect under the situation of bottom parts 31c and can easily disconnect along line of weakness.

Fig. 4 a-d represents another kind of microfluidic device of the present invention.This microfluidic device is the variant of microfluidic device shown in Fig. 3 a and the 3b, and different to be that it disposes two-layer:

● top part 41a, have end face 41a ' and comprise the groove that is used to provide fluid passage 42, be used for the hole and the solution inlet cavity 44 of the precipitating reagent cavity 46 of the contiguous inlet of precipitating reagent separately 45, and be used for the slit 48a of line of weakness is provided and is used for providing the slit 50a of line of weakness at precipitating reagent 45 places that enter the mouth in cutout section 48.

● seal member also constitutes bottom parts, and is applied in the form of sealant and covers described groove 42 and hole 46,44.

In use, target molecule solution is fed to unshowned target molecule solution inlet by target molecule solution inlet cavity 44, and target molecule solution is with flow channel 42 and fill up crystallization channel section 49.Because the progressively expansion from crystallization channel section 49 to precipitating reagent channel cavity 46 will not be so target molecule solution will can flow into precipitating reagent cavity 46.Progressively expansion from crystallization channel section 49 to precipitating reagent cavity 46 forms limit ' b ', and its angle is approximately 90 degree.This angle b can be another angle, but preferably less than 120 degree.

Precipitating reagent is fed into precipitating reagent cavity 46, and target molecule solution and precipitating reagent contact each other near precipitating reagent enters the mouth 45 places or its, and precipitating reagent will be diffused in the target molecule solution, it finally can cause the target molecule crystallization when cultivating.

After target molecule solution has been full of crystallization channel section 49, disconnect by the device that disconnects in the line of weakness of cutout channel section 48 at the fluid connection between unshowned branching channel section and the crystallization channel section 49, make various precipitating reagents can not flow into crystallization channel section 49 on every side.In Fig. 4 b, arrow A 1 expression is pushed the parts of the device that comprises the branching channel section to disconnect the direction of this device along line of weakness.

In the time of along the line of weakness disconnecting apparatus of cutout in the channel section 48 or afterwards, can provide the sealant 51 shown in Fig. 4 c and 4d.In the embodiment illustrated, sealant 41b be flexible and when device when line of weakness disconnects, the sealing layer does not disconnect.Therefore, comprise that the parts of the device of branching channel section can be used to sealant is squeezed in the fluid passage 42, so that crystallization channel section 49 and branching channel section are separated.In a selective embodiment, when along the line of weakness disconnecting apparatus, sealant also will disconnect.

After device is cultivated a period of time, be enough to make the precipitating reagent of desired amount to be diffused in the target molecule solution in the crystallization channel section during this period of time, so comprising the parts of the device that precipitates cavity 46 can disconnect along the line of weakness that provides in the cutout channel section 50, make precipitating reagent can not flow into the crystallization channel section again, and the while target molecule can not flow into the precipitating reagent in the precipitating reagent cavity again.After this, under the situation that does not have further loss target molecule risk, can continue to cultivate.

The parts that comprise the device of precipitating reagent cavity 46 disconnect with the similar manner that parts with the aforesaid device that comprises the branching channel section disconnect.In Fig. 4 b, arrow A 2 expressions are pushed the parts of the device that comprises precipitating reagent cavity 46 to disconnect the direction of this device along line of weakness.Sealant 51 provides as mentioned above.

Fig. 5 a-b represents the another kind of microfluidic device of the present invention, and this microfluidic device is the variant of microfluidic device shown in Fig. 4 a-d, and different be to be used to provide the slit 58a of line of weakness to have flat bottom branch.This device is equipped with 2 layers:

● top part 51a, this top part comprise the groove 52 that is used to provide fluid passage, are used for the hole and the solution inlet cavity 54 of the unshowned precipitating reagent cavity of the contiguous inlet of precipitating reagent separately, and the slit 58a that is used for providing in cutout section 58 line of weakness.

● seal member 51b also constitutes bottom parts, and applies with the form of sealant and to cover described groove and hole 54.

In Fig. 5 b, device seals crystallization channel section 59 along line of weakness disconnection and sealant from unshowned branching channel section.

Fig. 6 a is the figure of microfluidic device top part of the present invention.Fig. 6 b is the amplification sectional view of Fig. 6 a.This top part comprises that 64 and four of target molecule solution inlets pass through groove 67,68,69 precipitating reagents inlets 66 connected to one another, and this groove is used to provide fluid passage.Being equipped with groove 67,68,69 makes them to provide branching channel section 67, cutout channel section 68 and crystallization channel section 69 respectively.As can be seen, the groove 67 of branching channel section is greater than the groove 68 of cutout channel section, and the groove 68 of cutout channel section is again greater than the groove 69 of crystallization channel section.When the number of crystallization channel section was low, such structure was special expectation, added accurately little but sufficient quantity target molecule solution because be difficult to target molecule solution inlet.By this structure, target molecule solution will be introduced into the crystallization channel section owing to capillary force, and have the adequate space of excessive target molecule solution in the branching channel section.This device also comprises the excessive bag 63 of extra fluid, and it can be used for further collecting target molecule solution.

The top part of microfluidic device shown in Fig. 6 a and the 6b can be fixed to unshowned bottom parts such as simple film or plate, thereby fluid passage is provided and whole microfluidic device is provided.

Fig. 7 a-b is respectively first parts of microfluidic device of the present invention and complete microfluidic device.

The microfluidic device of Fig. 7 a-b comprises first parts of microfluidic device 71a, this microfluidic device have unshowned first with the relative groove face with groove 77,78,79, described groove 77,78,79 is used to provide branching channel section 77, cutout channel section 78 and crystallization channel section 79.Described groove face also is equipped with at the cavity between the crystallization channel section 79 73 separately.This microfluidic device also comprises and comprises many cover 71b to small part removable hood 71c.The first parts 71a also comprises through hole and the many through holes that is used for providing to fluid passage precipitating reagent inlet 76 that is used for providing to fluid passage target molecule solution inlet 74.Cover 71b is fixed on the groove face of the first parts 71a, enters the mouth 74,76 and separately channel section 77,78,79 separately to provide.

Removable hood 71c is applied on the groove 79 and along this groove sealing so that the crystallization channel section 79 of fluid passage to be provided.

Removable hood 71c extends the first parts 71a, is provided for shifting out separately the flange 71d that covers 71c separately.

Removable hood 71c releasably is connected to each other along pipeline 71e.

Fig. 8 is the schematic diagram of microfluidic device of the present invention, has wherein shown channel part and their contacts to each other.This microfluidic device comprises target molecule solution inlet 84 and more is used for the air opening/target molecule taphole 84a of excessive target molecule solution.In principle, described air opening/target molecule taphole 84a can be by the excessive export substitution of one or more air, and the excessive outlet of described air makes the air in the passage overflow with target molecule solution filling channel section.

Described microfluidic device also comprises 12 precipitating reagents inlet 86, and it is by branching channel section 87, cutout channel section 88 and 12 crystallization channel sections 89 and molecular solution 84 fluid connections that enter the mouth.As can be seen, cutout channel section 88 also partly constitutes the cutout channel section, but in this example in order to simplify, it only is called as " cutout channel section ".Air opening/target molecule taphole 84a is by interface channel section 87a and cutout channel section 88 fluid connections.This cutout channel section 88 can make the target molecule solution that has entered crystallization channel section 89 will can not flow back into cutout channel section 88 with respect to 89 displacements of crystallization channel section.Yet 89 pairs of target molecule solution of crystallization channel section should have the big capillary pulling power than interface channel part 87a, make target molecule solution will mainly flow into crystallization channel section 89 and be filled up to them that excessive subsequently target molecule solution will flow into interface channel section 87a.Alternatively or with displacement make up, cutout channel section 88 can be set to have the capillary force littler than crystallization passage.

Microfluidic device also comprises exhaust openings 81, it is closed during loading target molecule solution, for example close, and this exhaust openings 81 after being filled, is opened in the crystallization passage to discharge excessive target molecule solution and to participate in providing cutout in the cutout passage 88 simultaneously by stopper or floss hole.

In use, target molecule solution is joined target molecule solution inlet 84, and precipitating reagent is applied to precipitating reagent inlet 84a.Described target molecule solution flows into branching channel section 87, further flows into cutout channel section 88, and flows into 12 crystallization channel sections 89.When crystallization channel section 89 had been filled, excessive target molecule solution will flow among the interface channel section 87a and randomly flow into air opening/target molecule taphole 84a.If there is any target molecule solution to stay in the cutout channel section 88, it can be discharged by exhaust openings 81.

Fig. 9 represents the microfluidic device of second aspect present invention.This microfluidic device comprises first parts of microfluidic device 91a, this microfluidic device have unshowned first with relative groove face and the paired through hole that is used for providing respectively target molecule solution inlet 94 and precipitating reagent inlet 94.Paired inlet 94 connects by crystallization channel section 99, and this crystallization channel section 99 is provided by the groove in the first parts 91a groove face, and these first parts 91a tegmentum parts 91b covers.Cover 91b comprises and manyly movably covers 91c to small part.Described cover 91b is fixed to the groove face of the first parts 91a, with provide separately inlet 94 and crystallization channel section 99 separately.

Described removable hood 91c is applied on the groove 99 and along this groove sealing so that the crystallization channel section 99 of fluid passage to be provided.Removable hood 91c extends first parts 9 to be provided for the shifting out flange that covers 91c separately separately.

Removable hood 91c releasably is connected to each other along pipeline 91e.

Figure 10 is the top view of another microfluidic device of the present invention.This microfluidic device comprises target molecule solution inlet 104 and fluid passage, and this fluid passage comprises branching channel section 107, unshowned cutout channel section and crystallization channel section 109.For each crystallization channel section 109, this device comprises precipitating reagent inlet 106.Unshowned cutout channel section can be preferably provided by the groove 109 of adjacent grooves 107 at least in part, for example by providing lower capillary force to these sections or by alternate manner as herein described.

Microfluidic device also comprises removable hood 101c, and it for example can be aforesaid.Each removable hood 101c can comprise flange 101d separately, and it does not combine with underlying surface, and each lid 101c can move by the tearing among the described 101d of flange separately.

Claims (104)

1. microfluidic device that is used to promote the target molecule crystallization, described device comprises having end face, the solid structure of opposed bottom surface and at least one fluid passage, described fluid passage comprises target molecule solution inlet and at least two precipitating reagent inlets, this target molecule solution inlet is by described fluid passage and each precipitating reagent inlet fluid connection, described fluid passage comprises the branching channel section of contiguous described target molecule solution inlet, the crystallization channel section of the contiguous described inlet of precipitating reagent separately and be arranged on described branching channel section and described each crystallization channel section between the cutout channel section, described fluid passage is branched off into from 1 to X at described branching channel section, wherein X is the number of crystallization channel section, and described cutout channel section comprises cutout setting, and this cutout setting can disconnect the fluid connection between described channel section of branching separately and the crystallization channel section.

2. microfluidic device according to claim 1, wherein, described cutout be set to stop the separately form of the difference between the channel section, perhaps be the form of the difference of solid structure that described cutout channel section is provided, this cutout channel section is selected from a) branching channel section and b) at least a in the crystallization channel section of the contiguous described channel section that stops separately.

3. according to each described microfluidic device among the claim 1-2, wherein, described cutout is set at least a in the following form: a) capillary cutout, at least one cutout channel section wherein is set provides capillary force with the liquid in described passage, the capillary force that provides to liquid by described contiguous crystallization channel section is provided this capillary force, b) passage displacement, wherein the cutout passage of at least a portion is with respect at least one vertical displacement in branching channel section and the crystallization channel section, and c) line of weakness cutout, wherein said solid structure are equipped with the line of weakness of the fluid passage at least one cutout channel section of intersection.

4. microfluidic device according to claim 3, wherein, at least one cutout channel section is set provides capillary force with the liquid in described passage, this capillary force is lower than the capillary force that the crystallization channel section by described vicinity provides to liquid, and described cutout channel section is equipped with by the capillary of its geometry and/or surface tension setting and arrheas.

5. according to each described microfluidic device in the above claim, the wall that wherein said fluid passage comprises around passage and passage length extends, in at least one cutout channel section neutralization at least in the part around it, the surface tension of described wall is lower than the surface tension of the adjacent wall surface in the contiguous crystallization channel section, thereby cutout setting is provided.

6. microfluidic device according to claim 5, wherein at least one cutout channel section, in major part around it at least, such as at least 60%, at least 80%, at least 90%, at least basically all its around in, the surface tension of described wall is lower than the surface tension of the adjacent wall surface in the contiguous crystallization channel section, thereby cutout setting is provided.

7. according to each described microfluidic device among the claim 5-6, wherein at least a portion around at least one cutout channel section neutralizes it, the surface tension of described wall significantly is lower than the surface tension of the adjacent wall surface in contiguous crystallization channel section and the contiguous branching channel section at least one, such as low at least 5 dynes per centimeter, at least 10 dynes per centimeter, at least 20 dynes per centimeter, preferably, in at least one cutout channel section, in at least a portion around it, the surface tension of described wall is less than 73 dynes per centimeter, such as less than 60 dynes per centimeter, such as being the 10-55 dynes per centimeter.

8. according to each described microfluidic device among the claim 5-7, wherein said fluid passage has the polygonal cross-section shape on four limits that bottom surface, relative end face and two sides are provided at its cutout channel section at least, preferably, at least one surface tension in side and bottom surface and the end face is less than 73 dynes per centimeter, such as less than 60 dynes per centimeter, such as being the 10-55 dynes per centimeter.

9. according to each described microfluidic device in the above claim, the wall that wherein said fluid passage comprises around passage and passage length extends, in at least one of branching channel section and crystallization channel section, in at least a portion around it, the surface tension of described wall is at least 60 dynes per centimeter, such as at least 73 dynes per centimeter, such as at least 75 dynes per centimeter, such as at least 80 dynes per centimeter.

10. according to each described microfluidic device in the above claim, the wall that wherein said fluid passage comprises around passage and passage length extends, described fluid passage at least one of branching channel section and crystallization channel section has the polygonal cross-section shape on four limits that bottom surface, relative end face and two sides are provided, preferably, at least the surface tension of bottom surface and side is at least 60 dynes per centimeter, such as at least 73 dynes per centimeter, such as at least 75 dynes per centimeter, such as at least 80 dynes per centimeter.

11. microfluidic device according to claim 4, wherein said cutout channel section is equipped with by the capillary of its geometry setting and arrheas, and the smallest cross-sectional size of the passage of edge cutout channel section is greater than the smallest cross-sectional size along the passage of at least one in contiguous branching channel section and the crystallization channel section.

12. according to each described microfluidic device in claim 4 and 11, the area of section of wherein said cutout channel section is greater than at least one area of section in contiguous branching channel section and the crystallization channel section.

13. according to each described microfluidic device among the claim 11-12, wherein said fluid passage is included in the cataclysm of at least one sectional dimension in the cutout channel section of closely adjacent adjacent branching channel section and crystallization channel section one or both of.

14. according to each described microfluidic device in the above claim, wherein the area of section of at least one cutout channel section is greater than the area of section of contiguous crystallization channel section.

15. microfluidic device according to claim 14, the wall that wherein said fluid passage comprises around the passage and passage length extends, the larger cross-sectional area of described at least one cutout channel section are by at least along the progressively displacement formation of the wall of the parameatal major part in the described cutout channel section of next-door neighbour's crystallization channel section.

16. according to each described microfluidic device among the claim 14-15, wherein said fluid passage has the polygonal cross-section shape on four limits that bottom surface, relative end face and two sides are provided, preferably, the larger cross-sectional area of described at least one cutout channel section by the bottom surface at least of described cutout channel section and side with respect to contiguous crystallization channel section progressively displacement form.

17., wherein form precipitous displacement along the passage described progressively displacement of the wall of major part on every side in contiguous cutout channel section and the crystallization channel section at least according to each described microfluidic device among the claim 14-16.

18. according to each described microfluidic device among the claim 14-17, wherein form along the described progressively displacement of the wall of major part on every side of passage in the cutout channel section of next-door neighbour's crystallization channel section at least and have 120 degree or littler angles, such as 100 degree or littler angles, such as 90 degree or littler angles, such as the edge at 80 degree or littler angle.

19. according to each described microfluidic device among the claim 15-18, wherein by the near area of section of crystallization channel section greatly at least 10% near the larger cross-sectional area of at least one cutout channel section that the progressively displacement of major part forms around the passage in the described cutout channel section at least, such as big by 30% at least, big by 50% at least, big by 100% at least, big by 150% at least, big by 200% at least, big by 500% at least, big by 1000% at least.

20. microfluidic device according to claim 3, wherein at least one cutout channel section is provided by the passage displacement, wherein said cutout channel section is shifted onto the contiguous at least crystallization channel section of a ratio and preferably also has the high position of at least a portion branching channel section, wherein said higher position is for the end face of device, more preferably, at least one cutout channel section is displaced to a level with respect to the crystallization channel section, thereby when device remains on the horizontal plane, prevent that liquid from flowing back into the branching channel section from the crystallization channel section.

21. according to each described microfluidic device in the above claim, wherein said solid structure is equipped with the line of weakness of the fluid passage at least one cutout channel section of intersection, described line of weakness is intersected whole device, preferably, described line of weakness intersect simultaneously two or more the cutout channel sections, such as all cutout channel sections.

22. microfluidic device according to claim 21, wherein said line of weakness are to intersect the form in slit of whole device at least one of end face and bottom surface, this slit is preferably shaped to a kind of in V-arrangement and the U-shaped.

23. microfluidic device according to claim 22, the degree of depth in wherein said slit is at least 10% of a microfluidic device thickness, such as for the microfluidic device thickness in contiguous slit at least 15%, 30-90%, 35-80%, wherein the thickness of microfluidic device is defined as the distance between the relative bottom surface with it of end face.

24. according to each described microfluidic device in the above claim, each described two or more precipitating reagent inlets and precipitating reagent cavity fluid connection wherein are set, wherein, for each precipitating reagent inlet, described device preferably includes at least one precipitating reagent cavity.

25. according to each described microfluidic device in the above claim, wherein said device comprises the target molecule solution inlet cavity that is used for the target molecule solution feed is gone into target molecule solution inlet.

26. according to each described microfluidic device in the above claim, wherein said microfluidic device is equal to by two or more or the parts of different materials are made, these parts preferably are connected with each other by bonding, welding and/or mechanical grip.

27. microfluidic device according to claim 26, wherein said microfluidic device comprise have first and have the crystallization channel section that is used for providing at least fluid passage groove groove face at least the first parts and many at least one covers that movably covers to small part are provided, it has second and groove face separately, and wherein said movably lid is applied on the described groove and along this groove and seals so that the crystallization channel section of described fluid passage to be provided at least.

28. microfluidic device according to claim 27, wherein said first parts have at least one through hole or depression of being used to provide the target molecule solution inlet that leads to fluid passage.

29. according to each described microfluidic device among the claim 27-28, wherein said first parts have at least one through hole or depression of being used to provide the precipitating reagent inlet that leads to fluid passage.

30. according to each described microfluidic device among the claim 27-29, wherein said removable hood extends first parts, described removable hood preferably extends first parts to be provided for shifting out separately the flange of lid separately.

31. according to each described microfluidic device among the claim 27-30, wherein said removable hood comprises the flange that is not adhered on first parts separately, and this flange is set makes will to cover separately from crystallization channel section separately by the tearing in the described flange and shift out.

32. according to each described microfluidic device among the claim 27-31, wherein with described removable hood along separately groove by bonding and/or welded seal to the groove face of first parts.

33. according to each described microfluidic device among the claim 27-32, wherein said removable hood releasably is connected with each other.

34. according to each described microfluidic device among the claim 27-33, wherein said microfluidic device comprises three or more removable hoods, preferably, described microfluidic device comprises the removable hood that covers each described crystallization channel section.

35. according to each described microfluidic device among the claim 27-34, wherein said first parts are made by the material that is selected from glass, pottery, metal, semi-conducting material and polymer, described semi-conducting material for example is silicon, germanium and GaAs, and the Young's modulus of described polymer is 0.2GPa at least, such as 0.5GPa at least, such as 1GPa at least, described first parts preferably are transparent substantially.

36. according to each described microfluidic device among the claim 27-35, wherein said at least one cover is made by paper tinsel such as polymer foil or metal forming.

37. microfluidic device according to claim 36 wherein puts on described paper tinsel on the groove face of first parts, and is bonding or be welded on the groove face along groove, and partly or entirely is cut into pieces so that described removable hood to be provided.

38. according to each described microfluidic device in the above claim, wherein said microfluidic device comprises at least one by the parts that the material that is selected from glass, pottery, metal, semi-conducting material and polymer is made, and described semi-conducting material for example is silicon, germanium and GaAs.

39. according to the described method of claim 38, wherein said microfluidic device comprises the parts that at least one is made by polymeric material, described polymeric material is preferably the polymer of injection moldable, such as being selected from following polymer: acrylonitrile-butadiene-styrene copolymer, Merlon, dimethyl silicone polymer (PDMS), polyethylene (PE), polymethyl methacrylate (PMMA), polymethylpentene, polypropylene, polystyrene, polysulfones, polytetrafluoroethylene (PTFE), polyurethanes (PU), polyvinyl chloride (PVC), poly-inclined to one side 1,1-dichloroethylene (PVDC), poly-inclined to one side 1, the 1-difluoroethylene, styrene-propene acid copolymer (styrene-acryl copolymers), polyisoprene, polybutadiene, polychlorobutadiene, polyisobutene, poly-(s-B-S), siloxanes, epoxy resin, polyether block amide, polyester, acronitrile-butadiene-styrene (ABS), acrylic acid, celluloid, cellulose acetate, ethane-acetic acid ethyenyl ester (EVA), ethylene-vinyl alcohol (EVAL), fluoroplastics, polyacetals (POM), polyacrylate (acrylic acid), polyacrylonitrile (PAN), polyamide (PA), polyamide-imides (PAI), PAEK (PAEK), polybutadiene (PBD), polybutene (PB), polybutylene terephthalate (PBT), PET (PET), poly terephthalic acid hexamethylene dimethyl ester (PCT), polyketone (PK), polyester/polythene/polyethene, polyether-ether-ketone (PEEK), PEI (PEI), polyether sulfone (PES), polychlorostyrene ethene (polyethylenechlorinates) (PEC), polyimides (PI), PLA (PLA), polymethylpentene (PMP), polyphenylene oxide (PPO), polyphenylene sulfide (PPS), polyphthalamide (PPA) and composition thereof, preferably, at least one parts of described device are transparent, are used for optical check.

40. according to each described microfluidic device among the claim 26-39, at least one in the wherein said parts comprises the line of weakness of slit form.

41. according to each described microfluidic device among the claim 26-39, wherein said microfluidic device is made by bottom parts and top part at least, in bottom parts and the top part at least one comprises the groove that is used to provide fluid passage, and at least one in bottom parts and the top part comprises hole that is used for target molecule solution inlet cavity and the hole/depression that is used for the precipitating reagent cavity, and described bottom parts and described top part preferably are essentially rigidity.

42. according to the described microfluidic device of claim 41, wherein said microfluidic device is made by bottom parts and top part, described bottom parts and top part are fixed to one another by bonding, welding and/or mechanical grip, thereby form the fluid passage between the parts.

43. according to the described microfluidic device of claim 41, wherein said microfluidic device is made by the seal member of bottom parts, top part and sealant form at least, top part comprises the groove that is used to provide fluid passage, the hole that randomly is used for target molecule solution inlet cavity, and the hole/depression that is used for the precipitating reagent cavity, described seal member is clipped in and makes seal member cover described groove and described hole between top part and the bottom parts, and the described seal member of described base supports.

44. according to each described microfluidic device among the claim 41-43, wherein said bottom parts is the form of common plate basically, is preferably made by glass or polymer.

45. according to each described microfluidic device among the claim 43-44, wherein said bottom parts supports sealant at least in a zone, wherein the sealing layer covers the groove of top part to form the crystallization channel section, preferably described top part comprises the line of weakness of transcrystallization channel section, described line of weakness is preferably the part of the top part that extends bottom parts, and device can described line of weakness disconnect in the situation lower edge that does not disconnect bottom thus.

46. according to each described microfluidic device among the claim 26-39, wherein said microfluidic device is made by the seal member of bottom parts and sealant form at least, described top part comprises and is used to provide the groove of fluid passage and is used for target molecule solution inlet cavity and the hole of precipitating reagent inlet cavity that described seal member is connected with described top part makes described seal member cover described groove and described hole.

47. according to the described microfluidic device of claim 46, wherein said top part comprises the line of weakness of transcrystallization channel section, thereby device can disconnect along described line of weakness.

48. according to each described microfluidic device among the claim 43-47, the Shore A hardness of wherein said seal member is 90 or littler, is 20-70 such as Shore A hardness.

49. according to each described microfluidic device among the claim 34-48, wherein said seal member is made by polymeric material, this polymeric material is preferably elastomer, more preferably is selected from thermoplastic elastomer (TPE) and rubber.

50. according to each described microfluidic device among the claim 43-48, wherein said seal member is the form of common plate basically.

51. according to each described microfluidic device in the above claim, wherein said target molecule solution inlet cavity is the cavity form in described end face, described cavity is by described target molecule solution inlet and described branching channel section fluid connection, and described target molecule solution inlet cavity preferably has the conical in shape of the inlet from end face to target molecule solution.

52. according to the described microfluidic device of claim 51, the volume of wherein said target molecule solution inlet cavity is at least the cumulative volume of crystallization channel section, such as being at least 2 times, at least 1.5 times, at least 1.3 times, at least 1.2 times, at least 1.1 times of crystallization channel section volume, for example 0.05-20 μ l, 0.1-10 μ l, 0.2-5 μ l, 0.5-3 μ l.

53. according to each described microfluidic device in the above claim, at least one in the wherein said precipitating reagent cavity is semi-closed cavity form, described semi-closed precipitating reagent cavity is preferably used the precipitating reagent prefilled.

54. according to each described microfluidic device in the above claim, in the wherein said precipitating reagent cavity at least one is the inlet cavity form in described end face, described cavity is by described precipitating reagent inlet and described crystallization channel section fluid connection, and described precipitating reagent inlet cavity preferably has the conical in shape of the inlet from end face to target molecule solution.

55. according to each described microfluidic device in the above claim, wherein at least one crystallization channel section has the capillary plug that is arranged on or is close to the precipitating reagent inlet, this capillary plug preferably is preferably precipitous expansion by the progressively expansion from the crystallization channel section to the precipitating reagent cavity and provides, and preferably all the crystallization channel sections have the capillary plug that is arranged on or is close to their contiguous precipitating reagent inlet.

56. according to the described microfluidic device of claim 51, wherein progressively enlarging from the crystallization channel section to the precipitating reagent cavity forms and has 120 degree or littler angles, such as 100 degree or littler angles, such as 90 degree or littler angles, such as the edge at 80 degree or littler angle.

57. according to each described microfluidic device among claim 24 and the 53-56, the volume of wherein said precipitating reagent cavity is 0.01-10 μ l, such as 0.02-5 μ l, such as 0.5-2 μ l, such as 0.7-1.3 μ l.

58. according to each described microfluidic device in the above claim, wherein, except the inlet cavity with randomly precipitating reagent cavity and/or the line of weakness, described end face and opposed bottom surface are essentially smooth.

59. according to each described microfluidic device in the above claim, wherein said fluid passage comprises X crystallization channel section, wherein X is 2-1000, such as 2-400,4-100,8-48,8-24.

60. according to each described microfluidic device in the above claim, wherein said fluid passage has capillary dimensions on the major part of its length, described fluid passage is preferably in that branching channel section and crystallization channel section have capillary dimensions at least.

61. according to each described microfluidic device in the above claim, wherein said fluid passage has at least one less than 1000 μ m on the major part of its length, sectional dimension such as 10-250 μ m, 20-100 μ m, preferably, described fluid passage has at least one less than 1000 μ m at branching channel section and crystallization channel section at least, such as the sectional dimension of 10-250 μ m, 20-100 μ m.

62. according to each described microfluidic device in the above claim, wherein at least at described crystallization channel section, preferably on its whole length, described fluid passage has the polygonal cross-section shape on four limits, the polygonal cross-section shape on this four limit provides bottom surface, relative end face and two sides, the polygonal cross-section shape on described four limits is preferably trapezoidal, and these two trapezoidal sides are the 90-120 degree with respect to the angle of bottom surface, such as 90-110 degree, 90-100 degree, 90-95 degree.

63. according to the described microfluidic device of claim 62, wherein said fluid passage has the cross sectional shape that has width and height at the crystallization channel section at least, described height is the average distance between bottom surface and the end face, and described width is two average distances between the side, described width and be preferably 10-300 μ m with highly being mutually independent.

64. according to each described microfluidic device in the above claim, wherein said branching channel section comprises two or more branching sites.

65. according to each described microfluidic device in the above claim, the length of wherein said cutout channel section is 1-10000 μ m, such as 5-5000 μ m, 10-1000 μ m, 20-200 μ m.

66. according to each described microfluidic device in the above claim, wherein said cutout is set to intersect the form of line of weakness of fluid passage, the length of described cutout channel section is up to about 10000 μ m, such as being up to about 4000 μ m, being up to about 3000 μ m, being up to about 1000 μ m, being up to about 1000 μ m, being up to about 100 μ m, 1-50 μ m, it is defined as the length of the fluid passage that is intersected by line of weakness, and wherein the width of line of weakness and end face are measured abreast.

67. according to each described microfluidic device in the above claim, wherein said cutout is set to the capillary plug form in fluid passage, the length of described cutout channel section is up to about 10mm, such as 1-10000 μ m, 10-5000 μ m, 20-2000 μ m.

68. according to each described microfluidic device in the above claim, the length of wherein said crystallization channel section is 10-50000 μ m, such as 20-30000 μ m, 40-15000 μ m, 100-10000 μ m.

69. according to each described microfluidic device in the above claim, wherein said crystallization channel section is substantially parallel each other.

70. according to each described microfluidic device in the above claim, wherein said precipitating reagent cavity minimum range to each other is 0.1mm at least, such as 0.5-5mm, 1-3mm, 1.5-2.5mm.

71. according to each described microfluidic device in the above claim, wherein said device also comprises the line of weakness that is arranged in the described solid structure, the described crystallization channel section of the contiguous described inlet of precipitating reagent separately of this line of weakness intersection.

72. according to each described microfluidic device in the above claim, wherein said device also comprises the film of at least one the described crystallization channel section that mixes the described inlet of precipitating reagent separately of preferred vicinity.

73. according to the described microfluidic device of claim 72, wherein said film is a filter membrane, this filter membrane prevents that target molecule is diffused in the precipitating reagent that contacts with precipitating reagent inlet liquid, described film is preferably the perforated membrane of being made by one or more materials, described material is selected from fibrous material, such as melt jet glass fibre, spunbond synthetic fibers (for example nylon, polyester, polyethylene, polypropylene or polyether sulfone), semisynthetic fibre, regenerated fiber and inorfil and mixture; And foamable polymer, more preferably described film is a porous celluloid.

74. according to each described microfluidic device in the above claim, itself and target molecule solution make up, described target molecule solution also randomly comprises one or more washing agent in the aqueous solution.

75. microfluidic device described according to claim 74 and the combination of target molecule solution, wherein said cutout is set to capillary cutout form, at least one cutout channel section wherein is set provides capillary force with the target molecule solution in described passage, the capillary force that provides to target molecule solution by described contiguous crystallization channel section is provided this capillary force.

76. according to each microfluidic device described and combination of target molecule solution among the claim 74-75, the wall that wherein said fluid passage comprises around passage and passage length extends, in at least one cutout channel section and at least a portion around it, described wall has the surface tension lower than target molecule solution surface tension.

77. according to each microfluidic device described and combination of target molecule solution among the claim 74-76, the wall that wherein said fluid passage comprises around passage and passage length extends, in at least one crystallization channel section and at least a portion around it, described wall has the surface tension higher than target molecule solution surface tension.

78. according to each microfluidic device described and combination of target molecule solution among the claim 74-77, the wall that wherein said fluid passage comprises around passage and passage length extends, at least the described fluid passage in a crystallization channel section has the polygonal cross-section shape on four limits that bottom surface, relative end face and two sides are provided, preferably, described at least bottom surface and side have the surface tension higher than the surface tension of target molecule solution.

79. according to each microfluidic device described and combination of target molecule solution among the claim 74-78, the wall that wherein said fluid passage comprises around passage and passage length extends, described branching channel section and on every side to small part, described wall has the surface tension higher than the surface tension of target molecule solution.

80. according to each microfluidic device described and combination of target molecule solution among the claim 74-79, wherein said fluid passage has capillary dimensions on the major part of its length, preferably the described fluid passage in branching channel section and crystallization channel section at least has capillary dimensions, and this capillary dimensions is with respect to the target molecule measured in solution.

81. a method of utilizing each described microfluidic device in the above claim to promote the target molecule crystallization from target molecule solution, described method comprises:

Target molecule solution v) is provided, with the target molecule solution feed to target molecule solution inlet and make it be full of the crystallization channel section;

At least a precipitating reagent vi) is provided, and this precipitating reagent is set makes itself and at least one precipitating reagent inlet fluid connection;

Precipitating reagent is diffused in the target molecule solution of crystallization channel section; With

Viii) disconnect the fluid connection between the described crystallization of the target molecule separately channel section.

82. the method for 1 described promotion target molecule crystallization according to Claim 8, wherein said target molecule solution is the solution of at least a target molecule, described target molecule is selected from: protein, nucleic acid, nucleic acid analog, carbohydrate, lipid, more preferably be selected from 500 dalton or greater than 500 daltonian protein, strand and double-stranded DNA, RNA, PNA and LNA, and drug candidate.

83. the method for each described promotion target molecule crystallization among the 1-82 according to Claim 8, wherein said target molecule solution are fed directly to target molecule solution inlet, or by target molecule solution being applied in the target molecule solution inlet cavity.

84. the method for each described promotion target molecule crystallization among the 1-83 is wherein gone into the precipitating reagent cavity with the precipitating reagent prefilled according to Claim 8.

85. the method for each described promotion target molecule crystallization among the 1-83 according to Claim 8, wherein be provided with precipitating reagent come by precipitating reagent inlet separately be applied in the hole of containing precipitating reagent and with separately precipitating reagent inlet fluid connection.

86. the method for each described promotion target molecule crystallization among the 1-83 according to Claim 8, wherein be provided with precipitating reagent come by precipitating reagent is fed in the described device the precipitating reagent cavity and with separately precipitating reagent inlet fluid connection.

87. the method for each described promotion target molecule crystallization among the 1-86 according to Claim 8, wherein the fluid connection between the described crystallization of target molecule separately channel section is because the interior capillary force of fluid passage or because the displacement of cutout channel section and automatic disconnection.

88. the method for each described promotion target molecule crystallization among the 1-87 according to Claim 8, wherein, by disconnecting described device and disconnect fluid connection between the described crystallization of target molecule separately channel section along intersecting the line of weakness of cutout channel section.

89. the method for each described promotion target molecule crystallization among the 1-88 wherein, makes precipitating reagent be diffused into the time that keeps expectation in the target molecule crystallization channel section according to Claim 8, stops the fluid connection between precipitating reagent and the target molecule crystallization passage subsequently.

90. the method for each described promotion target molecule crystallization among the 1-89 according to Claim 8, this method also comprises the inlet that seals at least one crystallization channel section, and all inlets of preferably sealed crystallization channel section evaporate thus avoiding.

91., wherein, seal one or more inlets by following one or more modes according to the method for the described promotion target molecule of claim 90 crystallization:

A) by adding wax, come sealed entry such as paraffin or Tissuemat E; With

B) by the fixed latch element, such as making as polymer sheet or sheet glass by polymer or glass

Blocking element, this blocking element is preferably transparent.

Cultivate microfluidic device and crystal is formed and/or growth 92. the method for each described promotion target molecule crystallization among the 1-91 according to Claim 8, this method also comprise.

93. the method for each described promotion target molecule crystallization among the 1-92 according to Claim 8, this method also comprise from one or more crystallization channel sections, collect form/crystal that grows up to, preferably collect crystal by following one or more modes:

A) by at least two parts in the disconnecting apparatus separately forming the parts of crystallization channel section, and shift out crystal;

B) bottom parts or the top part of the crystallization channel section by optionally shifting out one or more selections, and shift out crystal; With

C) by from the crystallization channel section, drawing or extrude crystal.

94. microfluidic device that is used to promote the target molecule crystallization, described device comprise have first with second relative and the solid structure of at least one fluid passage, described fluid passage comprises target molecule solution inlet and at least one precipitating reagent inlet, described target molecule solution inlet is by described fluid passage and described precipitating reagent inlet fluid connection, described fluid passage comprises the crystallization channel section of contiguous described precipitating reagent inlet, described microfluidic device comprises at least the first parts that have first and have the groove face of at least one groove that is used to provide described crystallization channel section, and provide at least one cover to the small part removable hood, described lid has second and groove face, wherein described removable hood is applied on the described groove and is sealed to the crystallization channel section of described fluid passage along this groove.

95. according to the described microfluidic device of claim 94, wherein said first parts have at least one through hole or depression of being used to provide the target molecule solution inlet that leads to fluid passage.

96. according to each described microfluidic device among the claim 94-95, wherein said first parts have at least one through hole or depression of being used to provide the precipitating reagent inlet that leads to fluid passage.

97. according to each described microfluidic device among the claim 94-96, wherein said removable hood extends first parts, described removable hood preferably extends first parts to be provided for moving the flange of described removable hood.

98. according to each described microfluidic device among the claim 94-97, wherein said removable hood comprises the flange that is not adhered to first parts, and this flange is set makes to shift out lid from the crystallization channel section by the tearing in the described flange.

99. according to each described microfluidic device among the claim 94-98, wherein said removable hood along groove by bonding and/or welded seal to the groove face of first parts.

100. according to each described microfluidic device among the claim 94-99, wherein said microfluidic device comprises two or more fluid passages, this fluid passage comprises the crystallization channel section, and link to each other with the precipitating reagent inlet with target molecule solution inlet separately, described each crystallization channel section is preferably covered by removable hood, and this lid randomly releasably links to each other each other.

101. according to each described microfluidic device among the claim 94-100, wherein said microfluidic device comprises three or more removable hoods.

102. according to each described microfluidic device among the claim 94-101, wherein said first parts are made by the material that is selected from glass, pottery, metal, semi-conducting material and polymer, described semi-conducting material is such as being silicon, germanium and GaAs, and the Young's modulus of described polymer is 0.2GPa at least, such as 0.5GPa at least, 1GPa at least, described first parts are preferably substantial transparent.

103. according to each described microfluidic device among the claim 94-102, wherein said at least one cover is made by paper tinsel such as polymer foil or metal forming.

104. according to the described microfluidic device of claim 103, wherein described paper tinsel is applied on the groove face of described first parts, bonding or be welded on the groove face along groove, and partly or entirely be cut into pieces so that described removable hood to be provided.

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