CN104178423B - A kind of active somatic cell culture apparatus based on microflow control technique - Google Patents
A kind of active somatic cell culture apparatus based on microflow control technique Download PDFInfo
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Abstract
The invention discloses a kind of active somatic cell culture apparatus based on microflow control technique, comprise micro-fluidic cultivation chipset, swivel joint, rotating machine, micrurgy module and Flow Control pump, described swivel joint butts up against below described micro-fluidic cultivation chipset, described rotating machine is positioned at below described swivel joint, described rotating machine is provided with rotation axis, described rotation axis passes and stretches out described swivel joint, described rotating machine is fixedly connected with described micro-fluidic cultivation chipset by described rotation axis, described micrurgy module is connected with described micro-fluidic cultivation chipset, described Flow Control pump is connected with described micro-fluidic cultivation chipset by described swivel joint.The external sequence of passing through that apparatus of the present invention can carry out dozens of cell/embryo is simultaneously cultivated and to target surrounding environment and intravital micro-intervention operation, and can realize changing nutrient solution when not changing culture dish, apparatus of the present invention are rational in infrastructure, practical.
Description
Technical field
The present invention relates to a kind of active somatic cell culture apparatus based on microflow control technique.
Background technology
Cell is minimum vital movement unit, difficult to the viviperception of cell, but meaning is huge.The many multidate informations in life generation and growth course can be obtained to the vivo observation of embryo, be significant equally.Had to 2012 and be born all over the world more than 4,000,000 tube babies, and with the speed per year over 100000 in increase; The whole world has every year carried out more than in vitro fertilization (IVF) cycle of 1,000,000 examples; In the U.S., in ooecium slurry, the cycle life of microinjection (ICSI) accounts for 57.5% of full IVF cycle life.Latest survey shows, China's Infertile couples has also accounted for 15% of couple at child-bearing age's sum, at present, the medical expense in a conventional IVF cycle is about the tens thousand of left and right of Renminbi, if introduce ICSI to treat, expense also can increase, and wherein main cost all concentrates on reagent, the consumptive material consumption of (general 3-5 days) in culturing process.And at present, this field mainly adopts traditional manual manipulation method, need at different times, target to be proceeded in multiple different culture dish, technological operation very complicated, require high to operator's state of the art, repeatedly jump operation causes embryo's residence time in external environment longer, the disadvantageous effect such as easily to pollute.Based on above-mentioned present situation and drawback, develop a integrated, automatization, the multichannel manipulation in vitro for cell and body early embryo and sequential culture, micrurgy experimental system, there is society, economy, scientific research and ethical value widely.
Summary of the invention
Goal of the invention: technical problem to be solved by this invention is to provide a kind of active somatic cell culture apparatus based on microflow control technique, the external sequence of passing through that this device can realize dozens of cell/embryo is simultaneously cultivated, micro-intervention and Real-Time Monitoring operation, and can realize changing nutrient solution when not changing culture dish.
Summary of the invention: for solving the problems of the technologies described above, the technology used in the present invention means are:
A kind of active somatic cell culture apparatus based on microflow control technique, comprise micro-fluidic cultivation chipset, swivel joint, rotating machine, micrurgy module and Flow Control pump, described swivel joint butts up against below described micro-fluidic cultivation chipset, described rotating machine is positioned at below described swivel joint, described rotating machine is provided with rotation axis, described rotation axis passes and stretches out described swivel joint, described rotating machine is fixedly connected with described micro-fluidic cultivation chipset by described rotation axis, described micrurgy module is connected with described micro-fluidic cultivation chipset, described Flow Control pump is connected with described micro-fluidic cultivation chipset by described swivel joint,
Wherein, described micro-fluidic cultivation chipset comprises top cover, culture layer group, Flow Control layer I, Flow Control layer II and sealing bottom successively, described top cover is provided with multiple passage mounting, the edge of described top cover is provided with draw-in groove I, the center of described culture layer group is circular area of knockout, the edge of described circular area of knockout is provided with the main culture channel that multiple and circular area of knockout is communicated with, the described main culture channel the other end with catch runner and be connected, described in catch runner away from the port of center of circle end be runner mouth; Described Flow Control layer I lower surface center is provided with cylindrical protrusions, cylindrical protrusions is hollow structure, the internal surface of cylindrical protrusions is provided with internal thread, described Flow Control layer I lower surface is provided with annular channel I around cylindrical protrusions excircle, described annular channel I is communicated with multiple Flow Control runner I, the end of described Flow Control runner I is runner mouth I, and described runner mouth I is communicated with the main culture channel of culture layer group, and the edge of described Flow Control layer I is provided with multiple circle and gets through hole; Described Flow Control layer II center is provided with downgoing flow passage I, described Flow Control layer II lower surface is provided with annular channel II around downgoing flow passage I excircle, described annular channel II is communicated with many Flow Control runner II, the end of described Flow Control runner II is runner mouth II, described runner mouth II and the circle of Flow Control layer I get through that the runner mouth of hole and culture layer group is mutually corresponding to be communicated with, and described cylindrical protrusions stretches in downgoing flow passage I; Described sealing bottom edge is provided with draw-in groove II, and described draw-in groove II connects with the draw-in groove I of top cover is mutually corresponding, and described sealing bottom center is provided with downgoing flow passage II, and described downgoing flow passage I is inserted in downgoing flow passage II;
Wherein, described swivel joint is provided with three layers of concentric tube wall from outside to inside successively, be respectively tube wall I, tube wall II and tube wall III, described rotating machine is provided with the externally threaded rotation axis of band, described rotation axis passes swivel joint and stretches out swivel joint, the described cylindrical protrusions of micro-fluidic cultivation chip cooperatively interacts with the rotation axis projection of described rotating machine and is rotatably connected, the outer wall of described tube wall I and downgoing flow passage II connects, the outer wall of described tube wall II and downgoing flow passage I connects, the outer wall of described tube wall III and cylindrical protrusions connects, described tube wall I, two annular channels are formed successively from outside to inside between tube wall II and tube wall III, the Flow Control runner II of described micro-fluidic cultivation chipset is communicated with the annular channel between described swivel joint tube wall I and tube wall II, the Flow Control runner I of described micro-fluidic cultivation chipset is communicated with the annular channel between described swivel joint tube wall II and tube wall III, described swivel joint is also provided with the outflow keyholed back plate road of both ends open, one of them outflow keyholed back plate road is communicated with the annular channel between tube wall I and tube wall II, another outflow keyholed back plate road is communicated with the annular channel between tube wall II and tube wall III, described outflow keyholed back plate road is connected with described Flow Control pump,
Wherein, described micrurgy module comprises stepper-motor I, stepper-motor II, guide rail I, guide rail II, steady arm, needle holder, microneedle and micro-injection pump, described stepper-motor I is positioned on described guide rail I, described steady arm is driven by stepper-motor I and moves vertically, described guide rail II is fixedly connected with described stepper-motor I, described stepper-motor II is positioned on described guide rail II, described needle holder is driven by stepper-motor II and moves vertically, one end of described needle holder is connected with micro-injection pump by flexible pipe, its relative the other end is provided with pin permanent seat, described microneedle is sealedly connected on pin permanent seat, described steady arm is positioned at the syringe needle place of described microneedle, the syringe needle of described microneedle can move forward and backward in steady arm, the front end of described steady arm is provided with projection II.
Further preferably, described top cover also comprises micrurgy lid, and the edge of described micrurgy lid is provided with draw-in groove, and the center of described micrurgy lid is circular area of knockout, and the outer edge of circular area of knockout is a circle long strip shape area of knockout
Further preferably, described passage mounting is circular arc.
Further preferably, described micro-fluidic cultivation chipset adopts the casting of PMMA or PDMS material or etching to be prepared from.
Further preferably, described tube wall I, tube wall II and tube wall III are stainless material and are prepared from.
Further preferably, be equipped with application of anti-leakage layers in described tube wall I, tube wall II and tube wall III, described application of anti-leakage layers annular is affixed on inside tube wall.
Further preferably, described application of anti-leakage layers adopts graphite carbon, silicon carbide, wolfram varbide or stupalith to be prepared from.
Beneficial effect: compared to prior art, the external sequence of passing through that active somatic cell culture apparatus based on microflow control technique of the present invention can carry out dozens of cell/embryo is simultaneously cultivated and to target surrounding environment and intravital micro-intervention operation, as cell (ooecium slurry) interior microinjection, embryonic cell biopsy, foreign gene cell transfecting, cell paracrine intracellular signaling is studied, embryonic cell differentiation mechanism research etc., and can realize changing nutrient solution when not changing culture dish, apparatus of the present invention are rational in infrastructure, practical, overcome and need target to be proceeded to the loaded down with trivial details technological operation in multiple different culture dish at different times, and then avoid and cause embryo's residence time in external environment longer due to jump operation repeatedly thus situation about polluting embryo occurs.
Accompanying drawing explanation
Fig. 1 is the structural representation of micro-fluidic cultivation chipset entirety in apparatus of the present invention;
Fig. 2 is the structural representation of swivel joint in apparatus of the present invention;
The structural representation that in Fig. 3 apparatus of the present invention, rotating machine and swivel joint cooperatively interact;
Fig. 4 is 20 the micro-fluidic culture channel schematic diagram formed after apparatus of the present invention micro-fluidic cultivation chipset assembling bonding;
Fig. 5 is the structural representation of each micro-fluidic culture channel;
Fig. 6 is the structural representation that difference cultivates top cover;
Fig. 7 is the structural representation of micrurgy top cover;
Fig. 8 is the structural representation of culture layer I;
Fig. 9 is the structural representation of culture layer II;
Figure 10 is the structural representation of Flow Control layer I;
Figure 11 is the structural representation of Flow Control layer II;
Figure 12 is the structural representation of end sealing cover;
Figure 13 is the structural representation I of micrurgy module;
Figure 14 is the structural representation II of micrurgy module;
Figure 15 is the structural representation of apparatus of the present invention;
Figure 16 is the structural representation of micro-fluidic cultivation chipset split in apparatus of the present invention.
Embodiment
As shown in Fig. 1 ~ 16, active somatic cell culture apparatus based on microflow control technique of the present invention, comprise micro-fluidic cultivation chipset 1, swivel joint 2, rotating machine 3 and micrurgy module 4, rotating machine 3 is provided with the externally threaded rotation axis 5 of band, through hole is provided with in the middle of bottom swivel joint 2, rotation axis 5 passes this through hole and stretches out swivel joint 2, bottom swivel joint 2, rest part is sealed structure, the top rotary moveable of swivel joint 2 butts up against micro-fluidic cultivation chipset 1 bottom, rotating machine 3 is positioned at below swivel joint 2, rotating machine 3 is fixedly connected with micro-fluidic cultivation chipset 1 by rotation axis 5, micrurgy module 4 is connected with micro-fluidic cultivation chipset 1, apparatus of the present invention also comprise Flow Control pump 6, Flow Control pump 6 is connected with micro-fluidic cultivation chipset 1 by swivel joint 2,
As shown in figure 16, wherein, micro-fluidic cultivation chipset 1 is transparent by PMMA or PDMS etc., have certain flexible material to be formed by casting or lithography, and micro-fluidic cultivation chipset 1 comprises top cover 7, culture layer I8, culture layer II9, Flow Control layer I10, Flow Control layer II11 and sealing bottom 12 successively; Top cover 7 divides again difference to cultivate lid 601 and 602 two kinds, micrurgy lid, and difference is cultivated lid 601 and cultivated for the difference of different runner, and micrurgy lid 602 is for micrurgy;
As shown in Figure 6, difference cultivates the circular arc passage mounting 701 that lid 601 is provided with circle 20 decile, the center of circle that the center of circle and the difference of circular arc passage mounting 701 cultivate lid 601 overlaps, circular arc passage mounting 701 is corresponding with 20 main culture channel of culture layer group, difference is cultivated lid 601 and combine rear salable main culture channel with culture layer group, the edge that difference cultivates lid 601 is provided with fixed card slot I702, fixed card slot I702 and fixed card slot II1201 cooperatively interacts, and is combined by micro-fluidic cultivation chipset 1;
As shown in Figure 7, micrurgy lid 602 covers the circular area of knockout 801 that there are one and lid concentric in center, the outer edge of circular area of knockout 801 is the long strip shape area of knockout of circle 20 decile, long strip shape area of knockout overlaps with main culture channel is mutually corresponding, for providing the space of micrurgy needle movement, micrurgy lid 602 is horizontally rotated 9 °, can again seal main culture channel; The edge of micrurgy lid 602 is also provided with fixed card slot 802, and fixed card slot 802 and fixed card slot II1201 cooperatively interact, and are combined by micro-fluidic cultivation chipset 1;
Culture layer group is made up of culture layer I8 and culture layer II9, and one deck tips upside down on another layer, encapsulates, and forms 20 Flow Controls and cultivates unit;
The center of culture layer group is for being communicated with pond 902, being communicated with pond 902 is a circular area of knockout, top can be cultivated lid 601 by difference and be sealed, be communicated with the edge in pond 902 and be provided with multiple main culture channel 901 with being communicated with pond 902 and being communicated with, main culture channel 901 is divided in two parts, larger near the part being communicated with pond 902, less away from the part being communicated with pond 902, away from the port being communicated with pond 902 with catch runner 903 and be connected, the size of catching runner 903 can carry out processing adjustment according to the size of target cell/embryo, catching runner 903 away from center of circle end is runner mouth 904, runner mouth 904 is that a circle gets through hole, for being communicated with lower one deck chip,
As shown in Figure 5, main culture channel 901 is runner mouth 101 with the junction being communicated with pond 902, and runner mouth 101 for target cell/embryo being put into, or shifts out chip (docking with microinjection mouth); Main culture channel 901 is communicated with the runner mouth I1001 of Flow Control layer I, and runner mouth I1001 is for removing the granulosa cell around ovum or embryo; Main culture channel 901 is runner mouth 103 with the junction of catching runner 903, and runner mouth 103, for absorption, is caught and fixing Object of Development, to carry out micrurgy to it;
As shown in Figure 8, the center of culture layer I8 is circular area of knockout I, the edge of circular area of knockout I is provided with the long strip shape area of knockout I that multiple and circular area of knockout I is communicated with, long strip shape area of knockout I is divided in two parts, part near circular area of knockout I is larger, part away from circular area of knockout I is less, and the edge of culture layer I8 is provided with multiple circle and gets through hole 905;
As shown in Figure 9, the center of culture layer II9 is circular area of knockout II, the edge of circular area of knockout II connects the long strip shape area of knockout II that multiple and circular area of knockout II is communicated with, long strip shape area of knockout II is divided in two parts, part near circular area of knockout II is larger, part away from circular area of knockout II is less, long strip shape area of knockout II holds away from circular area of knockout II and catches runner 903 and be connected, and catches runner 903 and is provided with circle away from center of circle end and gets through hole 904 (namely catching the runner mouth 904 of runner); The circle of culture layer I8 is got through hole 905 and is got through with the circle of culture layer II9 that hole 904 is mutually corresponding to be communicated with;
As shown in Figure 10, Flow Control layer I10 chip monolithic molding, the below of culture layer group is positioned at during encapsulation, Flow Control layer I10 is provided with circle and gets through hole 1010 (runner mouth), this circle is got through hole 1010 and is got through hole 904 with the circle of culture layer group, 905 are connected, the circle of Flow Control layer I10 is got through hole 1010 and is also got through hole 1110 (runner mouth II) with the circle of Flow Control layer II11 and be connected, Flow Control layer I10 center is provided with the cylindrical protrusions 1004 of stretching out downwards, cylindrical protrusions 1004 is hollow structure, the internal surface of cylindrical protrusions 1004 is provided with internal thread, for being fixedly connected with the rotation that cooperatively interacts of the rotation axis 5 of below rotating machine 3, Flow Control layer I10 lower surface is provided with annular channel I1003 around cylindrical protrusions 1004 excircle, Flow Control runner I1002 is positioned at Flow Control layer I lower surface, size can as required---and mainly the size of target cell/embryo is carried out processing and is regulated, Flow Control runner I1002 one end is connected with annular channel I1003, the other end is runner mouth I1001, runner mouth I1001 is positioned at below the main culture channel 901 of culture layer group, and is communicated with main culture channel 901,
As shown in figure 11, Flow Control layer II11 chip monolithic molding, be positioned at below Flow Control layer I10 during encapsulation, Flow Control layer II11 center is provided with the downgoing flow passage I1101 stretched out downwards, downgoing flow passage I1101 adopts hollow structure, the cylindrical protrusions 1004 of Flow Control layer I10 can be inserted in it, and the runner formed between cylindrical protrusions 1004 with downgoing flow passage I1101 is connected with the annular channel between tube wall II1303 with the tube wall II1302 of swivel joint 2; Flow Control layer II11 lower surface is provided with annular channel II1102 around downgoing flow passage I1101 excircle, annular channel II1102 is communicated with many Flow Control runner II1103, the end away from center of circle end of Flow Control runner II1103 is that runner mouth II1110, Flow Control runner II1103 are communicated with runner mouth II1110 and annular channel II1102;
As shown in figure 12, sealing diagonal angle, bottom 12 edge is provided with two draw-in groove II1201, the draw-in groove (difference cultivates the draw-in groove I702 of lid 601 or the draw-in groove 802 of micrurgy lid 602) of draw-in groove II1201 and top cover 7 is mutually corresponding to be connected, the draw-in groove of draw-in groove II1201 and top cover 7 be combined with each other rotate stuck for whole micro-fluidic cultivation chipset 1 is tightly fixed together; Sealing bottom 12 center is provided with downgoing flow passage II1202, downgoing flow passage II1202 adopts hollow structure, downgoing flow passage I1101 is inserted in downgoing flow passage II1202, and the runner formed between downgoing flow passage I1101 with downgoing flow passage II1202 is connected with the annular channel between tube wall II1302 with the tube wall I1301 of swivel joint 2;
As shown in figures 2-3, swivel joint 2 location butts up against bottom micro-fluidic cultivation chipset 1, micro-fluidic cultivation chipset 1 rotary movable on swivel joint 2, and do not link with swivel joint 2, namely when swivel joint 2 is fixed time, micro-fluidic cultivation chipset 1 still can thereon by external force driven rotary, and swivel joint 2 is connected with Flow Control pump 6 with outflow keyholed back plate road 1306 by outflow keyholed back plate road 1305; Rotating machine 3 is positioned swivel joint 2 bottom, and rotating machine 3 is fixedly connected with micro-fluidic cultivation chipset 1 by rotation axis, thus micro-fluidic cultivation chipset 1 rotates together with rotating machine 3;
Swivel joint 2 is provided with three layers of concentric tube wall from outside to inside successively, be respectively tube wall I1301, tube wall II1302 and tube wall III1303, tube wall I1301, be positioned at swivel joint 2 outermost, enlarged area, top connects with the outer wall of sealing bottom 12 downgoing flow passage II1202, and be provided with application of anti-leakage layers in tube wall I1301, application of anti-leakage layers ring attaching is loaded on inside tube wall I1301, bottom is slightly narrow, and sidewall opening is connected with outflow keyholed back plate road 1306, tube wall II1302, be positioned at joint slightly inner side, the downgoing flow passage I1101 outer wall of enlarged area, top and Flow Control layer II11 connects, and have application of anti-leakage layers in tube wall II1302, application of anti-leakage layers ring attaching is loaded on inside tube wall, tube wall II1302 bottom is slightly narrow, and sidewall opening is connected with outflow keyholed back plate road 1305, have application of anti-leakage layers in tube wall III1303, application of anti-leakage layers ring attaching is loaded on inside tube wall III1303 and connects with cylindrical protrusions 1004 outer wall of Flow Control layer I10, rotating machine 3 is provided with the externally threaded rotation axis 5 of band, rotation axis 5 passes swivel joint 2 and stretches out swivel joint 2, the rotation that is used for cooperatively interacting with the cylindrical protrusions 1004 of micro-fluidic cultivation chipset 1 is fixedly connected with, thus by micro-fluidic cultivation chipset 1 together with rotating machine 3 secure bond, make micro-fluidic cultivation chipset 1 rotating machine 3 drive next play rotation, the outer wall of tube wall I1301 and downgoing flow passage II1202 connects, the outer wall of tube wall II1302 and downgoing flow passage I1101 connects, the outer wall of tube wall III1303 and cylindrical protrusions 1004 connects, tube wall I1301, two annular channels are formed successively from outside to inside between tube wall II1302 and tube wall III1303, the Flow Control runner II1103 of micro-fluidic cultivation chipset 1 is communicated with the annular channel between tube wall I1301 and tube wall II1302, the Flow Control runner I1002 of micro-fluidic cultivation chipset 1 is communicated with the annular channel between tube wall II1302 and tube wall III1303, swivel joint 2 is also provided with the outflow keyholed back plate road of both ends open, one of them outflow keyholed back plate road 1306 is communicated with the annular channel between tube wall I1301 and tube wall II1302, another outflow keyholed back plate road 1305 is communicated with the annular channel between tube wall II1302 and tube wall III1303, outflow keyholed back plate road 1305 is all connected with Flow Control pump 6 with outflow keyholed back plate road 1306, swivel joint 2 tube wall adopts stainless material, and application of anti-leakage layers can adopt the materials such as graphite carbon, silicon carbide, wolfram varbide, pottery to carry out leakproof, swabbing,
As shown in Figure 13 ~ 14, micrurgy module 4 comprises stepper-motor I1501, stepper-motor II1504, guide rail I1502, guide rail II1503, steady arm 1508, needle holder 1505, microneedle 1507 and syringe pump 1509, stepper-motor I1501 is positioned on guide rail I1502, guide rail II1503 is fixedly connected with stepper-motor I1501, stepper-motor II1504 is positioned on guide rail II1503, micrurgy module 4 entirety is driven by stepper-motor I1501, along guide rail I1502 tangential movement on an axis (Z axis), stepper-motor II1504 drives needle holder 1505 to do tangential movement along guide rail II1503 together, one end of needle holder 1505 is connected with syringe pump 1509 by flexible pipe, the relative the other end of needle holder 1505 is provided with pin permanent seat 1506, microneedle 1507 is sealedly connected on pin permanent seat 1506, needle holder 1505 and microneedle 1507 are interconnected, microneedle 1507 can disassemble from pin permanent seat 1506, change other entry needle, steady arm 1508 is positioned at the syringe needle place of microneedle 1507, the syringe needle of microneedle 1507 can move forward and backward in steady arm 1508, the front end of steady arm 1508 is provided with projection II, microneedle 1507 is combined by sleeve with steady arm 1508, cylinder inwall has lubricating substance, microneedle 1507 can seesaw along Z axis in sleeve, complete the puncture procedure of microinjection, steady arm 1508 docks (i.e. the projection II of steady arm 1508 insert main culture channel 901 front end) by front projection II with the main culture channel 901 of culture layer group in micro-fluidic cultivation chipset 1, complete location, drive steady arm 1508 to carry out axial motion by stepper-motor I1501 again, realize steady arm 1508 retreat operation.
Apparatus of the present invention carry out micro-manipulation by the fluid in Flow Control pump 6 pairs of runners in culturing process, to realize changing liquid and goal displacement in micro-fluidic cultivation chipset 1; In conjunction with rotating machine 3 and swivel joint 2, the flow-control operation in the rotary motion of micro-fluidic cultivation chipset 1 and rotary course can be realized; The micro-intervention operation to target can be realized again in conjunction with micrurgy module 4.
Obviously, above-described embodiment is only for example of the present invention is clearly described, and is not the restriction to embodiments of the present invention.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And these belong to spirit institute's apparent change of extending out of the present invention or change and are still among protection scope of the present invention.
Claims (7)
1. based on an active somatic cell culture apparatus for microflow control technique, it is characterized in that: comprise micro-fluidic cultivation chipset, swivel joint, rotating machine, micrurgy module and Flow Control pump; Described swivel joint butts up against below described micro-fluidic cultivation chipset, described rotating machine is positioned at below described swivel joint, described rotating machine is provided with rotation axis, described rotation axis passes and stretches out described swivel joint, described rotating machine is fixedly connected with described micro-fluidic cultivation chipset by described rotation axis, described micrurgy module is connected with described micro-fluidic cultivation chipset, and described Flow Control pump is connected with described micro-fluidic cultivation chipset by described swivel joint;
Wherein, described micro-fluidic cultivation chipset comprises top cover, culture layer group, Flow Control layer I, Flow Control layer II and sealing bottom successively, described top cover is provided with multiple passage mounting, the edge of described top cover is provided with draw-in groove I, the center of described culture layer group is circular area of knockout, the edge of described circular area of knockout is provided with the main culture channel that multiple and circular area of knockout is communicated with, the described main culture channel the other end with catch runner and be connected, described in catch runner away from the port of center of circle end be runner mouth; Described Flow Control layer I lower surface center is provided with cylindrical protrusions, cylindrical protrusions is hollow structure, the internal surface of cylindrical protrusions is provided with internal thread, described Flow Control layer I lower surface is provided with annular channel I around cylindrical protrusions excircle, described annular channel I is communicated with multiple Flow Control runner I, the end of described Flow Control runner I is runner mouth I, and described runner mouth I is communicated with the main culture channel of culture layer group, and the edge of described Flow Control layer I is provided with multiple circle and gets through hole; Described Flow Control layer II center is provided with downgoing flow passage I, described Flow Control layer II lower surface is provided with annular channel II around downgoing flow passage I excircle, described annular channel II is communicated with many Flow Control runner II, the end of described Flow Control runner II is runner mouth II, described runner mouth II and the circle of Flow Control layer I get through that the runner mouth of hole and culture layer group is mutually corresponding to be communicated with, and described cylindrical protrusions stretches in downgoing flow passage I; Described sealing bottom edge is provided with draw-in groove II, and described draw-in groove II connects with the draw-in groove I of top cover is mutually corresponding, and described sealing bottom center is provided with downgoing flow passage II, and described downgoing flow passage I is inserted in downgoing flow passage II;
Wherein, described swivel joint is provided with three layers of concentric tube wall from outside to inside successively, be respectively tube wall I, tube wall II and tube wall III, described rotating machine is provided with the externally threaded rotation axis of band, described rotation axis passes swivel joint and stretches out described swivel joint, the described cylindrical protrusions of micro-fluidic cultivation chip cooperatively interacts with the rotation axis projection of described rotating machine and is rotatably connected, the outer wall of described tube wall I and downgoing flow passage II connects, the outer wall of described tube wall II and downgoing flow passage I connects, the outer wall of described tube wall III and cylindrical protrusions connects, described tube wall I, two annular channels are formed successively from outside to inside between tube wall II and tube wall III, the Flow Control runner II of described micro-fluidic cultivation chipset is communicated with the annular channel between described swivel joint tube wall I and tube wall II, the Flow Control runner I of described micro-fluidic cultivation chipset is communicated with the annular channel between described swivel joint tube wall II and tube wall III, described swivel joint is also provided with the outflow keyholed back plate road of both ends open, one of them outflow keyholed back plate road is communicated with the annular channel between tube wall I and tube wall II, another outflow keyholed back plate road is communicated with the annular channel between tube wall II and tube wall III, described outflow keyholed back plate road is all connected with described Flow Control pump,
Wherein, described micrurgy module comprises stepper-motor I, stepper-motor II, guide rail I, guide rail II, steady arm, needle holder, microneedle and micro-injection pump, described stepper-motor I is positioned on described guide rail I, described steady arm is driven by stepper-motor I and moves vertically, described guide rail II is fixedly connected with described stepper-motor I, described stepper-motor II is positioned on described guide rail II, described needle holder is driven by stepper-motor II and moves vertically, one end of described needle holder is connected with micro-injection pump by flexible pipe, its relative the other end is provided with pin permanent seat, described microneedle is sealedly connected on pin permanent seat, described steady arm is positioned at the syringe needle place of described microneedle, the syringe needle of described microneedle can move forward and backward in steady arm, the front end of described steady arm is provided with projection II.
2. the active somatic cell culture apparatus based on microflow control technique according to claim 1, it is characterized in that: described top cover also comprises micrurgy lid, the edge of described micrurgy lid is provided with draw-in groove, the center of described micrurgy lid is circular area of knockout, and the outer edge of circular area of knockout is a circle long strip shape area of knockout.
3. the active somatic cell culture apparatus based on microflow control technique according to claim 1, is characterized in that: described passage mounting is circular arc.
4. the active somatic cell culture apparatus based on microflow control technique according to claim 1, is characterized in that: described micro-fluidic cultivation chipset adopts the casting of PMMA or PDMS material or etching to be prepared from.
5. the active somatic cell culture apparatus based on microflow control technique according to claim 1, is characterized in that: tube wall I, the tube wall II of described swivel joint and tube wall III are stainless material and are prepared from.
6. the active somatic cell culture apparatus based on microflow control technique according to claim 1, it is characterized in that: be equipped with application of anti-leakage layers in tube wall I, the tube wall II of described swivel joint and tube wall III, described application of anti-leakage layers annular is affixed on inside tube wall.
7. the active somatic cell culture apparatus based on microflow control technique according to claim 6, is characterized in that: described application of anti-leakage layers adopts graphite carbon, silicon carbide, wolfram varbide or stupalith to be prepared from.
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| CN105802832B (en) * | 2016-03-03 | 2018-06-15 | 河海大学常州校区 | The microinjection device and its operating method of batch microinjection can be carried out to cell |
| US11390836B2 (en) * | 2016-11-17 | 2022-07-19 | Cleveland State University | Chip platforms for microarray 3D bioprinting |
| CN108690805A (en) * | 2017-04-07 | 2018-10-23 | 清华大学 | A kind of automatic carry out cell culture metabolism and the online device and method collected or detect |
| CN107904167B (en) * | 2017-11-15 | 2023-06-06 | 中国航天员科研训练中心 | Automatic cell expansion culture equipment |
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