CN104630061B - Micro-fluidic solution concentration generation and culture detection chip - Google Patents
Micro-fluidic solution concentration generation and culture detection chip Download PDFInfo
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- CN104630061B CN104630061B CN201310547371.7A CN201310547371A CN104630061B CN 104630061 B CN104630061 B CN 104630061B CN 201310547371 A CN201310547371 A CN 201310547371A CN 104630061 B CN104630061 B CN 104630061B
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Abstract
The invention relates to a micro-fluidic solution concentration generation and culture detection chip. The chip integrates concentration generation, suspension culture channel and detection channel, and can obtain a specific concentration gradient as needed to meet diverse gradient concentration demands, a gradient generation unit occupies a small area, array formation can be easily realized, precise control of the flow velocity or long time balance is not needed, and the sample introduction is simple; suspension culture and detection of microbes can be simultaneously realized; and in addition, detection driving channels comprise at least two driving channels, so a culture solution can be detected multiple times in different time periods.
Description
Technical field
The invention belongs to micro-fluidic field, more particularly to a kind of micro-fluidic solution concentration generation and culture detection chip.
Background technology
In industrial microorganism fermentation production process, efficient microorganism conversion is to be together decided on by superior strain and optimal condition of culture.Traditional microbe to screen is mainly carried out in shaking flask, detects two key steps including microbial suspension culture and culture fluid.Carry out suspension culture by accessing each bacterial strain in shaking flask, then material detection is carried out to the culture fluid in shaking flask, so that it is determined that the performance of bacterial strain, select excellent bacterial strain.Every batch of screening often needs to be tested with tens of up to a hundred shaking flasks.Nonetheless, still far below bacterial strain quantity to be screened, this directly results in screening process high labor intensive to shaking flask screening quantity, and screening efficiency is low.In addition, screening process also can consume substantial amounts of culture medium and other reagent；Need to take larger culture and working place etc..Secondly, the purpose that could finally realize high yield is optimized to the condition of culture of superior strain.There is significant difference in different microorganisms condition of culture, not only the species and content of primary carbon source, nitrogen source, phosphorus source are required different, and to other micro constitutents, such as somatomedin, metallic element etc. such as vitamin also has diversified demand.Therefore, the optimization of condition of culture is needed to investigate the impact to growth of microorganism fermentation under multiple concentration conditions for the specific nutrition composition.Conventional training systern experiment is carried out generally in shaking flask, adds the nutritional labeling of culture fluid and specified quantitative one by one in shaking flask, places into shaking table vibration and cultivated after adding bacterium.Whole process not only needs to carry out the sample-adding operation of repeatability in a large number, but also a large amount of shaking flasks can be used, this needs to spend substantial amounts of manpower, sample-adding number of times is more simultaneously, microbiological contamination probability is higher, the consumption of culture fluid is also very big, and in addition experiment also needs to use multiple shaking tables, and whole experiment process efficiency is low, high cost, time length, need substantial amounts of equipment and space.These problems present in training systern experiment therefore in the urgent need to address.
Microflow control technique is that last century the nineties grow up in analytical chemistry field, and the function components and parts image set such as microchannel, Micropump, micro-valve, with microchannel network microstructure features, is become circuit the same by micro-processing technology, is integrated on chip material by it.Microflow control technique has high efficiency, due to structure small it is easy on chip once integrated tens of up to a hundred microorganism culturing units, culture fluid can be injected simultaneously into multiple culture units, conventional efficient is improved simultaneously；The consumption of culture fluid and other medicines also can be greatly decreased, thus reducing screening cost；The small volume of micro-fluidic chip, it is possible to reduce the usage amount of incubator, reduces requisite space and the equipment limit of microorganism culturing；Micro-fluidic chip can carry out batch machining production and pretreatment（As cleaning, sterilizing etc.）, become disposable test consumable, so not only can reduce the manufacturing cost of chip, it is possible to reduce the operation that test prepares, the final labor intensity reducing screening operation.
Microbial suspension culture can be carried out at present both at home and abroad and the micro-fluidic chip of culture fluid detection is also little, the chip of report also haves such problems as that flux is low, processed complex, versatility be not high, still has a segment distance from microbe to screen application.
Noo Li Jeon describes the micro-fluidic chip that a kind of pyramid can quickly form Concentraton gradient in patent WO200222264, this chip has 3 solution inlet, it is injected separately into low concentration, middle concentration and highly concentrated solution, after 9 grades of lateral network allocation mixing, mixed liquor flows out from 9 outlets, and each outlet solution concentration forms gradient.Input this chip with less kind of original concentration solution and be achieved with multiple Concentraton gradient, whole process achieves miniaturization and automatization, it is to avoid in routine operation, solution repeats the step added, and can save manpower, improves operating efficiency.But, the design of this chip depends on multilevel branch piping network, increase with concentration export volume, lateral network series also accordingly increases, and this can take larger chip area, is unfavorable for the integrated of other chip functions units of later stage, increased series also can improve injection pressure, increased injection flow speed control difficulty, and concentration occurs to be affected very big by injecting flow velocity, needs to carry out precise control to flow velocity.In addition, single group pyramid gradient generating unit produce solution gradient species also than relatively limited although can be combined by multiple Gradient Units, increase several gradient species, but species still than relatively limited it is difficult to meet diversified Concentraton gradient requirement of experiment, and more chip area can be taken.
Jian Liu describes a kind of micro-fluidic reaction array based on circulation mixing in patent US 2010/0104477.This array contains liquid mixing unit and the fluid containment structure such as micro-valve, Micropump of 400 square closures.Each liquid mixing unit can be injected to different liquids, be then spaced from each other each square closed cells with micro-valve by controlling successively in the pipeline on rectangular cells a line or two sides, start Micropump and carry out solution mixing in unit.This chip can be realized the injection of different liquids mass and mix, and operating efficiency is high, and chip occupying area is little, be realize on chip multiple-unit batch sample introduction with quick mix provide new approaches.However, each unit structure is identical in this reaction array, the solution concentration producing after each unit mixing identical it is impossible to the multiple solution concentration of generation that is directly used in mass, still cannot be directly used to diversified Concentraton gradient experiment.
Todd Thorsen in 2002, et al. report a kind of micro-fluidic optical comparable chip（Science 298, 580 (2002)； DOI: 10.1126/science.1076996）.This chip contains 256 reaction members, and each unit includes the chamber of a receiving antibacterial and the chamber of a receiving nitrite ion, has a micro-valve to be separated by between two chambers.After the completion of sample introduction, open micro-valve and so that antibacterial is contacted with nitrite ion and react, the fluorescence signal that detection produces is it may determine that whether antibacterial expresses specific protein.Although this chip detection flux is very high, make cell proliferation due to cannot be carried out suspension culture in chip, be only used for the screening of individual bacteria it is impossible to be used for diversified microbe to screen target, versatility is not high.
Nicolas Szita reports a kind of micro- reaction chip of multichannel micro-fluidic within 2005（DOI: 10.1039/b504243g）.4 reaction chambers of this integrated chip, have miniature stirring paddle be used for realizing microbial suspension culture, and are mounted with that two patch sensors realize the detection to culture fluid pH and dissolved oxygen content in each cavity.Although this chip is capable of microbial suspension culture, and detects pH and oxyty in culture fluid, because the volume of single reaction chamber is larger, approximate number hectolambda, simultaneously need to process miniature stirring paddle, integrated patch sensor, complex manufacturing technology, is difficult to chip unit quantity is greatly improved.
Chinese Patent No. CN201110316751.0 and No. CN201110142095.7 individually disclose a kind of micro-fluidic cell suspension culture chip, and the chip referring in this two pieces patent can realize the microorganism batch parallelization suspension culture of passages up to a hundred.However, this chip is only capable of determining the cell quantity in culture fluid by Cytometric method, lack culture fluid detection structure it is impossible to be measured to the concentration of special component in culture fluid.
In view of this it is necessary to provide one kind can obtain specific gradient concentration, and realize suspension culture and the micro-fluidic chip of analysis detection simultaneously.
Content of the invention
It is an object of the invention to provide a kind of micro-fluidic solution concentration occurs and culture detection chip, specific gradient concentration can be obtained, disclosure satisfy that various gradient concentration demand, and realize the suspension culture of microorganism or cell and the detection of special component in culture fluid simultaneously.
For achieving the above object, the present invention provides following technical scheme：
A kind of micro-fluidic solution concentration occurs and culture detection chip, including the culture layer being stacked, elastic diaphragm layer and driving layer, described elastic diaphragm layer is located at described culture layer and drives between layer, more than one culture detector unit is distributed with described culture layer, each culture detector unit includes the culture raceway groove of annular and the detection raceway groove being connected with described culture raceway groove, circulation is distributed with described driving layer drives raceway groove and detection to drive raceway groove, wherein, described circulation drives raceway groove to be located at the top of described culture raceway groove and become to intersect with described culture channel shape, and drive the culture fluid in described culture raceway groove to circulate；Described detection drives raceway groove to include at least two driving raceway grooves and be respectively positioned on the top of described detection raceway groove and become to intersect with described detection channel shape, 9th driving raceway groove is also distributed with described driving layer, described 9th driving raceway groove is formed in primary importance with described culture raceway groove and intersects, and this primary importance is divided into upper culture unit and lower culture unit by cultivating raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described culture detector unit also includes nitrite ion injection channel, this nitrite ion injection channel is communicated in described culture raceway groove or detection raceway groove, and described detection drives raceway groove to be located at described circulation and drives between raceway groove and nitrite ion injection channel.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described nitrite ion injection channel is Z-shape.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the 3rd driving raceway groove is also distributed with described driving layer, the 3rd driving raceway groove is formed with described nitrite ion injection channel and sense channel respectively and intersects.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the nitrite ion injection channel between described adjacent culture detector unit is connected.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the 5th driving raceway groove is also distributed with described driving layer, the nitrite ion injection channel formation between adjacent culture detector unit intersects the 5th driving raceway groove respectively.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, 4th driving raceway groove is also distributed with described driving layer, 4th driving raceway groove is located at the top of described culture raceway groove and becomes to intersect with described culture channel shape, and described 4th driving raceway groove is located at described circulation and drives between raceway groove and detection driving raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described culture detector unit also includes cleanout fluid injection channel, and this cleanout fluid injection channel is communicated in described culture raceway groove, and described cleanout fluid injection channel is located at the described 4th and drives between raceway groove and detection raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described cleanout fluid injection channel is communicated in the joint of described culture raceway groove and detection raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described culture detector unit also includes cleanout fluid injection channel, and this cleanout fluid injection channel is communicated in described detection raceway groove, and is located between adjacent two driving raceway grooves in described detection raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the cleanout fluid injection channel between described adjacent culture detector unit is connected.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the 7th driving raceway groove is also distributed with described driving layer, the cleanout fluid injection channel formation between adjacent culture detector unit intersects the 7th driving raceway groove respectively.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, the volume of the lower culture unit of each culture raceway groove described is different.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, between described culture raceway groove, be communicated with fluid pipe, described primary importance is pressed close in the junction of described fluid pipe and described culture raceway groove.
Preferably, occur in above-mentioned micro-fluidic solution concentration and cultivate in detection chip, described driving layer is additionally provided with the tenth driving raceway groove, the tenth driving raceway groove is in order to control the on or off of fluid pipe between adjacent culture raceway groove.
Compared with prior art, it is an advantage of the current invention that：The micro-fluidic solution concentration of the present invention occurs and concentration by culture detection chip occurs, suspension culture raceway groove and detection raceway groove are integrated on same chip, specific gradient concentration can be obtained on demand, disclosure satisfy that various gradient concentration demand, gradient generating unit area occupied is little, easily realize array, flow velocity precise control or long-time balance need not be carried out, sample introduction is simple；And realize suspension culture and the detection of microorganism simultaneously；Additionally, detection drives raceway groove to include at least two driving raceway grooves, may be implemented in different time sections and repeated detection is carried out to culture fluid.
Brief description
In order to be illustrated more clearly that the embodiment of the present application or technical scheme of the prior art, the accompanying drawing of required use in embodiment or description of the prior art will be briefly described below, apparently, drawings in the following description are only some embodiments described in the application, for those of ordinary skill in the art, on the premise of not paying creative work, can also obtain other accompanying drawings according to these accompanying drawings.
Fig. 1 show micro-fluidic solution concentration in preferred embodiment and the top view with culture detection chip occurs；
Fig. 2 show the sectional view in Fig. 1 along 1D；
Fig. 3 show micro-fluidic solution concentration in second embodiment of the invention and the top view with culture detection chip occurs；
Fig. 4 show micro-fluidic solution concentration in third embodiment of the invention and the top view with culture detection chip occurs；
Fig. 5 show micro-fluidic solution concentration in fourth embodiment of the invention and the top view with culture detection chip occurs.
Specific embodiment
The embodiment of the invention discloses a kind of micro-fluidic solution concentration occurs and culture detection chip, including the culture layer being stacked, elastic diaphragm layer and driving layer, described elastic diaphragm layer is located at described culture layer and drives between layer, more than one culture detector unit is distributed with described culture layer, each culture detector unit includes the culture raceway groove of annular and the detection raceway groove being connected with described culture raceway groove, circulation is distributed with described driving layer drives raceway groove and detection to drive raceway groove, wherein, described circulation drives raceway groove to be located at the top of described culture raceway groove and become to intersect with described culture channel shape, and drive the culture fluid in described culture raceway groove to circulate；Described detection drives raceway groove to include at least two driving raceway grooves and be respectively positioned on the top of described detection raceway groove and become to intersect with described detection channel shape, 9th driving raceway groove is also distributed with described driving layer, described 9th driving raceway groove is formed in primary importance with described culture raceway groove and intersects, and this primary importance is divided into upper culture unit and lower culture unit by cultivating raceway groove.
The micro-fluidic solution concentration of the present embodiment occurs chip can quickly produce multiple solution concentrations, and carries out microorganism culturing wherein.Chip need not add pyramid lateral network, and gradient generating unit area occupied is little, easily realizes array, flow velocity precise control or long-time balance need not be carried out, sample introduction is simple, can obtain specific gradient concentration on demand, disclosure satisfy that various gradient concentration demand.
The microfluidic microbe of the application is cultivated detection chip and can also be realized the microbial suspension culture of mass and the detection of culture fluid special component.Integrated chip element number is many, and up to up to a hundred units, and processing and fabricating is simple, need not install stirring paddle or integrated detection paster, be highly suitable for the rapid screening of a large amount of bacterial strains.
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is described in detail it is clear that described embodiment is only a part of embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, the every other embodiment that those of ordinary skill in the art are obtained on the premise of not making creative work, broadly fall into the scope of protection of the invention.
Fig. 1 show micro-fluidic solution concentration in first embodiment of the invention and the top view with culture detection chip occurs；Fig. 2 show the sectional view in Fig. 1 along 1D.
Shown in ginseng Fig. 1 and Fig. 2, micro-fluidic solution concentration occurs to include culture layer 11 with culture detection chip 10, culture layer 11 is at least formed by the combination of any one or more in high molecular polymer, hydrogel, silicon chip, quartz, glass and metal material, preferably, culture layer 11 is made up of polydimethylsiloxane.
Multiple culture detector units 111 are distributed with culture layer 11, culture detector unit 111 is shape, the unit of volume all same, and be arranged in parallel between unit side by side.Detection raceway groove 1112 and nitrite ion injection channel 1113 that the culture raceway groove 1111 of culture raceway groove 1111 and annular that culture detector unit 111 includes annular is connected, nitrite ion injection channel 1113 is communicated in detection raceway groove 1112.
Nitrite ion injection channel 1113 is arranged to Z-shaped that is to say, that nitrite ion injection channel 1113 is connected with detection raceway groove 1112 after bending twice, and the nitrite ion injection channel 1113 between adjacent culture detector unit 111 is connected.
It is communicated with fluid pipe 112 between culture detector unit 111, fluid pipe 112 includes multiple fluid pipe units 1121, each fluid pipe unit 1121 is communicated between adjacent culture raceway groove 1111, fluid pipe unit 1121 extend transversely and with culture raceway groove 1111 perpendicular, multiple fluid pipe units 1121 be stepped formula arrangement.
Culture detector unit 111 also includes cleanout fluid injection channel 1114, and this cleanout fluid injection channel 1114 is communicated in culture raceway groove 1111 and the joint of detection raceway groove 1112.
The lower end of all culture detector units 111 is communicated in fluid pipe 113, fluid pipe 113 and ft connection jointly.Solution can be injected simultaneously into in all culture detector units 111 by fluid pipe 113.
Will be readily apparent, all culture detector units 111 can be each independent unit, that is, cultivate the lower end of detector unit 111 and do not connect altogether, have independent import respectively, so can inject different solution in different culture detector units 111.
The top stacking of culture layer 11 is provided with elastic diaphragm layer 12, and elastic diaphragm layer 12 is formed by elastopolymer material it is preferred that elastic diaphragm layer 12 is made up of polydimethylsiloxane.
The top stacking of elastic diaphragm layer 12 is provided with driving layer 13, layer 13 is driven at least to be formed by the combination of any one or more in high molecular polymer, hydrogel, silicon chip, quartz, glass and metal material, preferably, layer 13 is driven to be made up of polydimethylsiloxane.
Drive and circulation driving raceway groove 131 is distributed with layer 13, circulation drives raceway groove 131 to be located at the top of culture raceway groove 1111 and intersect with cultivating raceway groove 1111 and being formed.
Circulation drives the two ends of raceway groove 131 to be connected with the external world, gases at high pressure are injected when driving to circulation in raceway groove 131, circulation drives the elastic diaphragm layer 12 under raceway groove 131 can bend downwards, block the culture raceway groove 1111 of elastic diaphragm layer 12 lower section, when removing gases at high pressure, elastic diaphragm layer 12 recovers, and the culture raceway groove 1111 of lower section connects, and this is micro-valve known to microfluidic art.
It is two or three parallel pipelines that circulation drives raceway groove 131, by pressurizeing successively by specific time sequence, can extrude the liquid one-way flowing in lower section culture raceway groove 1111, this is Micropump known to microfluidic art.
Circulation drives raceway groove 131 and cultivates circulation driving structure and its principle that raceway groove 1111 is formed, and discloses, the present embodiment repeats no more in Chinese Patent No. CN201110316751.0 and No. CN201110142095.7.
Drive and detection driving raceway groove 132 is also distributed with layer 13, detection drives raceway groove 132 to include two driving raceway grooves, it is respectively first and drive raceway groove 1321 and the second driving raceway groove 1322, first driving raceway groove 1321 and the second driving raceway groove 1322 are respectively positioned on the top of detection raceway groove 1112 and are intersected with detection raceway groove 1112 formation, drive raceway groove 131 principle the same with circulation, first drives raceway groove 1321 and second to drive raceway groove 1322 to form " micro-valve " in the infall with detection raceway groove 1112.
Drive and the 6th driving raceway groove 133 is also distributed with layer 13, the 6th drives raceway groove 133 to intersect the bottom end opening being located at culture detector unit 111（Culture fluid entrance）Place, intersects the top being located at fluid pipe 113, after culture fluid injects culture detector unit 111, can be passed through gases at high pressure, to realize the closing to culture detector unit 111 culture fluid porch in the 6th driving raceway groove 133.Will be readily apparent, in order to realize the closing to culture detector unit 111 entrance, also can pass through the other modes such as sealing and realize closure.
Drive and the 3rd driving raceway groove 134 is also distributed with layer 13, the 3rd driving raceway groove 134 is formed with nitrite ion injection channel 1113 and sense channel 1112 respectively and intersects.
In prior art, the separation between conventional each culture detector unit employs interdigitated pneumatic micro valve, drives the overlapping between layer and culture layer to need to be accurately positioned in two dimensions of horizontal plane.Preferably positioning is in longitudinal direction to interdigitated pneumatic micro valve（Culture unit bearing of trend）Upper two interdigitated micro-valves are located at the both sides of horizontal fluid pipeline respectively, longitudinal close-packed arrays but not overlapping with liquid channel, transversely interdigitated pneumatic micro valve will be intersected with longitudinal fluid pipeline, and interdigital length is as far as possible short simultaneously, reduce chip occupying area, improve integrated level.However, because, in chip manufacture manufacturing process, chip two-layer is directed at overlapping in two dimensions of horizontal plane and can there is error.In order to prevent processing the error impact chip use producing, need to increase the spacing of interdigitated pneumatic micro valve and interdigital length, this thereby necessarily increases the chip area of micro-valve occupancy, reduce the integrated level of chip.In addition, when there is slight difference because base material shrinkage factor is different in layers of chips figure, even can accurately be aligned in certain region, dimensional discrepancy between two layer patterns also can increase with alignment point distance and increase, lead to the volume of each reaction chamber inconsistent, impact detection accuracy, even unit are not used to detect completely.
Drive raceway groove 134 equally can realize the function of separating between unit using Z-shaped nitrite ion injection channel 1113 with reference to linear type the 3rd, and the required precision of each interlayer assembling is lower, levels overlapping only needs keeping parallelism in a dimension, need not in two dimensions of horizontal plane accurate alignment stack, reduce chip manufacturing difficulty, be favorably improved the quantity of chip unit.
In certain embodiments, drive the 5th driving raceway groove 135 that interdigitated can also be distributed with layer 13, nitrite ion injection channel 1113 formation between adjacent culture detector unit 111 intersects the 5th driving raceway groove 135 respectively.Specifically, 5th driving raceway groove 135 includes lateral extensions 1351 and longitudinal extension part 1352, wherein, lateral extensions 1351 horizontal expansion and with culture detector unit 111 square crossing, the sectional area of lateral extensions 1351 is smaller, will not form " micro-valve " with culture detector unit 111 in infall；Longitudinal extension part 1352 extends longitudinally and is communicated in lateral extensions 1351, longitudinal extension part 1352 is formed between adjacent culture detector unit 111 and is formed with nitrite ion injection channel 1113 and intersects, realize " micro-valve " function, when being passed through gases at high pressure in the 5th driving raceway groove 135, can achieve to the separation between culture detector unit 111, reduce the probability of fluid cross-contamination between unit.Longitudinal extension part 1352 forms intersection also at the entrance and exit of nitrite ion injection channel 1113, to realize " micro-valve " function it is easy to it is contemplated that also can realize sealing by modes such as sealings at the entrance and exit of nitrite ion injection channel 1113.
4th driving raceway groove 136 is also distributed with driving layer 13, the 4th driving raceway groove 136 intersects positioned at the top of culture raceway groove 1111 and with cultivating raceway groove 1111 and being formed, and the 4th drives raceway groove 136 to drive between raceway groove 132 positioned at circulation driving raceway groove 131 and detection.
In other embodiments, cleanout fluid injection channel 1114 also can be communicated in detection raceway groove 1112, and is located between adjacent two driving raceway grooves in detection raceway groove 1112.Specifically, cleanout fluid injection channel 1114 is located at first and drives between raceway groove 1321 and the second driving raceway groove 1322.In order to realize more preferable cleaning action, cleanout fluid injection channel 1114 is it can also be provided that multiple, for example drive between raceway groove 1322 in the first driving raceway groove 1321 and second and the joint of culture raceway groove 1111 and detection raceway groove 1112 is equipped with cleanout fluid injection channel 1114, the present invention is not intended to limit to this.
Cleanout fluid injection channel 1114 between adjacent culture detector unit 111 is connected.Drive and the 7th driving raceway groove 137 is also distributed with layer 13, cleanout fluid injection channel 1114 formation between adjacent culture detector unit 111 intersects the 7th driving raceway groove 137 respectively.The structure of the 7th driving raceway groove 137 drives raceway groove 135 identical with the 5th, all in order to realize cultivating the separation between detector unit 111.
Under normal circumstances, to in nitrite ion injection channel 1113, the reactant in nitrite ion injection channel 1113, detection raceway groove 1112 can be gone out pipeline by injection cleanout fluid, complete to clean, but, when detecting that the reactant in raceway groove 1112 is still difficult to rinse well, then need to be filled with gases at high pressure to the 4th driving raceway groove 136, inject cleanout fluid in cleanout fluid injection channel 1114, cleanout fluid is unidirectional to flow through detection raceway groove 1112, completes entirely to detect the cleaning of raceway groove 1112.As microbial suspension culture need to be proceeded after the completion of cleaning operation, then should drive in raceway groove 1321 to the 7th driving raceway groove 137, first and be filled with gases at high pressure, remove the 4th driving raceway groove 136 mesohigh gas, open Micropump and drive culture fluid to circulate, carry out suspension culture.
Drive and the 8th driving raceway groove 138 is also distributed with layer 13,8th drives raceway groove 138 to intersect at the end outlet being located at culture detector unit 111, after culture fluid injects culture detector unit 111, gases at high pressure can be passed through in the 8th driving raceway groove 138, to realize the closing to culture detector unit 111 exit.Will be readily apparent, in order to realize the closing to culture detector unit 111 outlet, also can pass through the other modes such as sealing and realize closure.
Drive and the 9th driving raceway groove 139 is also distributed with layer 13, the 9th driving raceway groove 139 is formed in primary importance A with culture raceway groove 1111 and intersects, this primary importance A is divided into upper culture unit 1115 and lower culture unit 1116 by cultivating raceway groove 1111.9th driving raceway groove 139 includes multiple driving trench cells 1391, each drives trench cells 1391 to intersect the top of close two race being located at adjacent culture raceway groove 1111, drive trench cells 1391 extend transversely and with culture raceway groove 1111 perpendicular, multiple driving trench cells 1391 be stepped formula arrangement.Try one's best primary importance A in the junction of fluid pipe unit 1121 and culture raceway groove 1111.
Drive and the tenth driving raceway groove 140 is also distributed with layer 13, fluid pipe unit 1121 formation between adjacent culture raceway groove 1111 intersects the tenth driving raceway groove 140 respectively, and forms micro-valve in infall, in order to control the on or off of fluid pipe unit 1121.Tenth driving raceway groove 140 is also formed with fluid pipe 113 and intersects, and form micro-valve in infall, when being passed through gases at high pressure in the tenth driving raceway groove 140, tenth drives raceway groove 140 to can achieve to the closing cultivating detector unit 111 lower ending opening, realizes the separation between culture detector unit 111 lower end simultaneously.
There is the operation logic with culture detection chip 10 in micro-fluidic solution concentration：First to injection solution A in all culture detector units 111 until being full of, then to the 6th driving raceway groove 133, 9th driving raceway groove 139 pressurizes, close fluid pipe 113 and the culture raceway groove 1111 that it passes through, and inject solution B in fluid pipe 112, solution B enters the pipeline being located between the 6th driving raceway groove 133 and the 9th driving raceway groove 139 in the culture raceway groove 1111 of annular closed, the solution A being simultaneously originally present in this section of tubing is rushed out, repressurization closes the tenth micro-valve driving raceway groove 140 control, there is solution A and solution B in so same culture detector unit 111 simultaneously, two kinds of solution are driven the micro-valve that raceway groove 139 controls to separate by the 9th.The part that the volume of solution B is closed by the micro-valve of the tenth driving raceway groove 140 and the 9th driving raceway groove 139 control cultivates the volume of detector unit 111.The volume of solution A is that the volume of entirely culture detector unit 111 deducts the volume shared by solution B.The first driving raceway groove 1321 is closed in pressurization.Open the micro-valve that the 9th driving raceway groove 139 controls, after starting the Micropump that circulation drives raceway groove 131 composition, solution A and solution B are cultivated in raceway groove 1111 in ring-type and circulated mixing.Because fluid pipe 112 can be arranged on diverse location, two neighboring culture raceway groove 1111 is connected, tenth driving raceway groove 140 and the 9th driving raceway groove 139 control the volume that the part of micro-valve closing cultivates unit 111 can change on request, therefore can produce multiple specific Concentraton gradient.Solution A, solution B can be the true solutions such as glucose solution, peptone solution, water or the suspension containing granules such as microorganisms.But solution A and solution B can be different solution or the different solution of same solution concentration.Microorganism with circulating current, carries out suspension growth in culture raceway groove 1111, and wherein part culture fluid can be deposited in detection raceway groove 1112.Before detection, nitrite ion is injected by nitrite ion injection channel 1113, pressurize the 3rd driving raceway groove 134, then it is passed through gas-pressurized in the second driving raceway groove 1322, and remove the gas-pressurized in the first driving raceway groove 1321, culture fluid is reacted with the nitrite ion in detection raceway groove 1112, and the optical signalling of generation is collected by extraneous optical detector, thus carrying out quantitation to the predetermined substance in culture fluid such as Phos, glucose etc..After the completion of reaction, it is passed through cleanout fluid, detergent line in cleanout fluid injection channel 1114 or nitrite ion injection channel 1113, in case detect next time.By above method, in different time sections, the culture fluid in culture detector unit 111 can be detected.
In second embodiment of the invention, micro-fluidic solution concentration occurs also can only arrange a culture detector unit with culture detection chip, shown in ginseng Fig. 3.
Shown in ginseng Fig. 4, in third embodiment of the invention, position in culture raceway groove for the adjustment fluid pipe 312, adjustment simultaneously the 9th drives raceway groove 339 and the position of the tenth driving raceway groove 340, it is possible to obtain isocratic concentration distribution.
Shown in ginseng Fig. 5, in fourth embodiment of the invention, position in culture raceway groove for the adjustment fluid pipe 412, adjustment simultaneously the 9th drives raceway groove 439 and the position of the tenth driving raceway groove 440, it is possible to obtain random concentration distribution.
It should be noted that, herein, such as first and second or the like relational terms are used merely to make a distinction an entity or operation with another entity or operation, and not necessarily require or imply there is any this actual relation or order between these entities or operation.And, term " inclusion ", "comprising" or its any other variant are intended to comprising of nonexcludability, so that including a series of process of key elements, method, article or equipment not only include those key elements, but also include other key elements of being not expressly set out, or also include for this process, method, article or the intrinsic key element of equipment.In the absence of more restrictions, it is not excluded that also there is other identical element in process, method, article or the equipment including described key element in the key element being limited by sentence "including a ...".
The above is only the specific embodiment of the application; it should be pointed out that for those skilled in the art, on the premise of without departing from the application principle; some improvements and modifications can also be made, these improvements and modifications also should be regarded as the protection domain of the application.
Claims (15)
1. a kind of micro-fluidic solution concentration occurs and culture detection chip, includes the culture layer being stacked, elastic diaphragm layer and drives layer, described elastic diaphragm layer be located between described culture layer and driving layer it is characterised in that：More than one culture detector unit is distributed with described culture layer, each culture detector unit includes the culture raceway groove of annular and the detection raceway groove being connected with described culture raceway groove, circulation is distributed with described driving layer drives raceway groove and detection to drive raceway groove, wherein, described circulation drives raceway groove to be located at the top of described culture raceway groove and become to intersect with described culture channel shape, and drives the culture fluid in described culture raceway groove to circulate；Described detection drives raceway groove to include at least two driving raceway grooves and be respectively positioned on the top of described detection raceway groove and become to intersect with described detection channel shape, 9th driving raceway groove is also distributed with described driving layer, described 9th driving raceway groove is formed in primary importance with described culture raceway groove and intersects, and this primary importance is divided into upper culture unit and lower culture unit by cultivating raceway groove.
2. micro-fluidic solution concentration according to claim 1 occur with culture detection chip it is characterised in that：Described culture detector unit also includes nitrite ion injection channel, and this nitrite ion injection channel is communicated in described culture raceway groove or detection raceway groove, and described detection drives raceway groove to be located at described circulation and drives between raceway groove and nitrite ion injection channel.
3. micro-fluidic solution concentration according to claim 2 occur with culture detection chip it is characterised in that：Described nitrite ion injection channel is Z-shape.
4. micro-fluidic solution concentration according to claim 3 occur with culture detection chip it is characterised in that：3rd driving raceway groove is also distributed with described driving layer, the 3rd driving raceway groove is formed with described nitrite ion injection channel and sense channel respectively and intersects.
5. the micro-fluidic solution concentration according to Claims 2 or 3 occur with culture detection chip it is characterised in that：Nitrite ion injection channel between described adjacent culture detector unit is connected.
6. micro-fluidic solution concentration according to claim 5 occur with culture detection chip it is characterised in that：5th driving raceway groove is also distributed with described driving layer, the nitrite ion injection channel formation between adjacent culture detector unit intersects the 5th driving raceway groove respectively.
7. micro-fluidic solution concentration according to claim 1 occur with culture detection chip it is characterised in that：4th driving raceway groove is also distributed with described driving layer, the 4th drives raceway groove to be located at the top of described culture raceway groove and become to intersect with described culture channel shape, and the described 4th drives raceway groove to drive raceway groove and detection to drive between raceway groove positioned at described circulation.
8. micro-fluidic solution concentration according to claim 7 occur with culture detection chip it is characterised in that：Described culture detector unit also includes cleanout fluid injection channel, and this cleanout fluid injection channel is communicated in described culture raceway groove, and described cleanout fluid injection channel is located at the described 4th and drives between raceway groove and detection raceway groove.
9. micro-fluidic solution concentration according to claim 8 occur with culture detection chip it is characterised in that：Described cleanout fluid injection channel is communicated in the joint of described culture raceway groove and detection raceway groove.
10. micro-fluidic solution concentration according to claim 7 occur with culture detection chip it is characterised in that：Described culture detector unit also includes cleanout fluid injection channel, and this cleanout fluid injection channel is communicated in described detection raceway groove, and is located between adjacent two driving raceway grooves in described detection raceway groove.
11. micro-fluidic solution concentrations according to claim 8 or claim 9 occur with culture detection chip it is characterised in that：Cleanout fluid injection channel between described adjacent culture detector unit is connected.
12. micro-fluidic solution concentrations according to claim 11 occur with culture detection chip it is characterised in that：7th driving raceway groove is also distributed with described driving layer, the cleanout fluid injection channel formation between adjacent culture detector unit intersects the 7th driving raceway groove respectively.
13. micro-fluidic solution concentrations according to claim 1 occur with culture detection chip it is characterised in that：The volume of the lower culture unit of each culture raceway groove described is different.
14. micro-fluidic solution concentrations according to claim 13 occur with culture detection chip it is characterised in that：It is communicated with fluid pipe, described fluid pipe presses close to described primary importance with the described junction cultivating raceway groove between described culture raceway groove.
15. micro-fluidic solution concentrations according to claim 14 occur with culture detection chip it is characterised in that：Tenth driving raceway groove is additionally provided with described driving layer, the tenth driving raceway groove is in order to control the on or off of fluid pipe between adjacent culture raceway groove.
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| CN201310547371.7A CN104630061B (en) | 2013-11-06 | 2013-11-06 | Micro-fluidic solution concentration generation and culture detection chip |
| PCT/CN2013/001451 WO2014082377A1 (en) | 2012-11-28 | 2013-11-27 | Microfluidic chip |
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| CN201310547371.7A CN104630061B (en) | 2013-11-06 | 2013-11-06 | Micro-fluidic solution concentration generation and culture detection chip |
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| WO2009139898A2 (en) * | 2008-05-16 | 2009-11-19 | President And Fellows Of Harvard College | Valves and other flow control in fluidic systems including microfluidic systems |
| CN102234614A (en) * | 2011-05-30 | 2011-11-09 | 中国科学院苏州纳米技术与纳米仿生研究所 | Micro-fluidic cell suspension culture chip and application thereof |
| CN102286358A (en) * | 2011-05-31 | 2011-12-21 | 中国科学院合肥物质科学研究院 | Microfluidic control chip for realizing PCR (Polymerase Chain Reaction) and real-time PCR virus quick detection device |
| CN102337207A (en) * | 2011-10-18 | 2012-02-01 | 中国科学院苏州纳米技术与纳米仿生研究所 | Microfluidic microbe two-dimensional suspension culture chip |
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| WO2009139898A2 (en) * | 2008-05-16 | 2009-11-19 | President And Fellows Of Harvard College | Valves and other flow control in fluidic systems including microfluidic systems |
| CN102234614A (en) * | 2011-05-30 | 2011-11-09 | 中国科学院苏州纳米技术与纳米仿生研究所 | Micro-fluidic cell suspension culture chip and application thereof |
| CN102286358A (en) * | 2011-05-31 | 2011-12-21 | 中国科学院合肥物质科学研究院 | Microfluidic control chip for realizing PCR (Polymerase Chain Reaction) and real-time PCR virus quick detection device |
| CN102337207A (en) * | 2011-10-18 | 2012-02-01 | 中国科学院苏州纳米技术与纳米仿生研究所 | Microfluidic microbe two-dimensional suspension culture chip |
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