Liquid drop micro-fluidic chip

Abstract

The utility model relates to the technical field of microfluidics, in particular to a droplet micro-fluidic chip. Comprising a main pipeline, a first pipeline and a second pipeline, the first pipeline and the second pipeline are respectively communicated with the main pipeline, and the first pipeline and the second pipeline are respectively positioned on two opposite sides of the main pipeline; the same liquid flows in the first pipeline and the second pipeline. The hydrogel microspheres loaded with the hiPSCs not only are beneficial to transfer and control of the hydrogel microspheres loaded with the hiPSCs, but also can load different types of cells in the shell and core space of the microspheres, thereby being beneficial to research on the interaction relationship between cells or organoid derived from the cells.

Description

Technical Field

[0001] This application relates to the field of microfluidics, and more particularly to a droplet microfluidic chip. Background Technology

[0002] Currently, matrix gels and alginates are widely used in organoid culture due to their high biocompatibility. Matrix gels are the most commonly used extracellular matrix material in organoid culture; they are liquid at 4°C and gradually solidify as the temperature rises. Due to their relatively low mechanical strength, matrix gels pose difficulties for the manipulation and transfer of microspheres. Alginates are widely used to support the growth of cell clumps / organoids because they are readily available and inexpensive, but they cannot provide attachment sites for single-cell growth, making them unsuitable for the culture of stem cell-derived organoids. However, in another strategy, this material can be used as a shell for hydrogels, and its concentration can be adjusted to regulate the shell's mechanical properties, facilitating automated operations such as microsphere transfer.

[0003] Because these two types of hydrogels have different cross-linking mechanisms, the preparation of core-shell structured hydrogel microspheres based on these two types of hydrogels has not yet been discovered. Utility Model Content

[0004] In this application, we propose a simple, rapid, and high-throughput method for preparing alginate-matrix-oil dual hydrogel microdroplets, thereby achieving high-throughput preparation of shell-core structured composite hydrogel microspheres. The preparation of these microspheres not only facilitates subsequent high-throughput culture and automated operations (such as microsphere transfer and oil phase removal) of stem cell-derived organoids, but also enables the study of cell-cell or cell-derived organoid interactions, providing reliable technical support for research in drug development, regenerative medicine, precision medicine, and other fields.

[0005] High-throughput generation of hydrogel microspheres has opened up opportunities for high-throughput, homogeneous culture of organoids. Currently, most hydrogel microspheres used for cell loading are solid microspheres prepared from a single hydrogel material or hollow hydrogel microcapsules formed via droplet microfluidics. These two forms of hydrogel microspheres / capsules are not universally suitable for most organoid cultures requiring 3D scaffold support or for the co-culture of different cell / organoid types. Currently, various organoids have been cultured using matrix gels; however, matrix gels have relatively low mechanical strength and are prone to depolymerization and collapse during long-term culture. To provide a stable 3D growth environment for organoids, we designed a droplet microfluidic chip and, for the first time, proposed the preparation of alginate-matrix-oil dual hydrogel microdroplets. Through curing under specific conditions, high-throughput fabrication of core-shell composite hydrogel microspheres was achieved.

[0006] To address the problems existing in the prior art, this application provides a droplet microfluidic chip.

[0007] The specific technical solution of this application is as follows:

[0008] A droplet microfluidic chip includes a main channel through which liquid flows, a first channel, and a second channel; the first channel and the second channel are respectively connected to the main channel and are located on opposite sides of the main channel; the same liquid flows in the first channel and the second channel.

[0009] A third and / or fourth pipe intersects and connects with the main pipe;

[0010] The third and / or fourth pipes are capable of dividing the continuously flowing liquid in the main pipe into droplets;

[0011] The third pipe and / or the fourth pipe are located downstream of the first pipe and the second pipe.

[0012] In one specific embodiment, a first connection is formed at the intersection of the first pipe and the main pipe, and a second connection is formed at the intersection of the second pipe and the main pipe; preferably, the first connection and the second connection coincide.

[0013] In one specific embodiment, the diameter of the main pipeline gradually increases from upstream to downstream.

[0014] In one specific embodiment, the diameters of the first pipe and the second pipe are equal or similar.

[0015] In one specific embodiment, a core liquid flows in the main pipe, a first liquid flows in the first pipe, and a second liquid flows in the second pipe; preferably, both the first liquid and the second liquid flow toward the interior of the main pipe; more preferably, the core liquid, the first liquid, and the second liquid merge at the first connection and / or the second connection to form a fifth liquid;

[0016] More preferably, the flow rate of the core liquid in the main pipeline is 1~20 μL / min; more preferably, the flow rate of the core liquid in the main pipeline is 4~10 μL / min.

[0017] In one specific embodiment, the fifth liquid flows in the main pipe.

[0018] In one specific embodiment, the flow rate of the first liquid is equal to or similar to the flow rate of the second liquid; preferably, the cross-sectional diameter and flow rate of the first liquid and the second liquid are equal to or similar.

[0019] More preferably, the flow rate of the first liquid in the first pipe is 1~10 μL / min; more preferably, the flow rate of the first liquid in the first pipe is 2~5 μL / min;

[0020] More preferably, the flow rate of the second liquid in the second pipe is 1~10 μL / min; more preferably, the flow rate of the second liquid in the second pipe is 2~5 μL / min.

[0021] In one specific embodiment, a third pipe and a fourth pipe are intersecting and connected on the main pipe;

[0022] Preferably, the third pipe and the fourth pipe are positioned opposite each other on both sides of the main pipe.

[0023] In one specific embodiment, the diameter of the third pipe is equal to or similar to the diameter of the fourth pipe; preferably, the diameters of both the third pipe and the fourth pipe are less than or equal to the diameter of the main pipe.

[0024] In one specific embodiment, the intersection of the third pipe and the main pipe forms a third connection, and the intersection of the fourth pipe and the main pipe forms a fourth connection; preferably, the third connection and the fourth connection coincide.

[0025] In one specific embodiment, a third liquid flows inside the third pipe, and a fourth liquid flows inside the fourth pipe. The third liquid and the fourth liquid are capable of shearing the liquid in the main pipe into droplets.

[0026] More preferably, the flow rate of the third liquid in the third pipe is 1~5 μL / min;

[0027] More preferably, the flow rate of the fourth liquid in the fourth pipe is 1~5 μL / min.

[0028] In one specific embodiment, a fifth pipe and / or a sixth pipe are connected to the main pipe;

[0029] Preferably, a fifth pipe and a sixth pipe are connected in an intersecting manner on the main pipe;

[0030] Preferably, the fifth pipe and the sixth pipe are arranged opposite each other on both sides of the main pipe;

[0031] More preferably, the fifth pipe and the sixth pipe are both located downstream of the third pipe and the fourth pipe.

[0032] In one specific embodiment, a sixth liquid flows inside both the fifth and sixth pipes.

[0033] In one specific embodiment, the chip further includes a first inlet pipe disposed upstream of the main pipe, through which liquid flows into the main pipe;

[0034] Preferably, the chip further includes a second inlet channel, which is connected to the first channel and the second channel;

[0035] Preferably, the chip further includes a third inlet channel, which is connected to the third channel and the fourth channel;

[0036] Preferably, the chip further includes a fourth inlet channel, which is connected to the fifth channel and the sixth channel.

[0037] Beneficial effects

[0038] The droplet microfluidic chip of this application consists of three cross-shaped channels. The core phase of the chip is a mixture of matrix gel and hiPSCs, and the outer shell phase is a mixture of sodium alginate and Ca-EDTA. Due to the low Reynolds number of the two hydrogel mixtures flowing through the chip channels, the core and outer shell phases form an immiscible laminar flow. Subsequently, at the second cross-shaped channel, the droplet is dissected by the oil phase to form a monodisperse droplet composed of an aqueous core (the matrix gel is liquid at 4°C) and a hydrogel shell. When the droplet passes through the third cross-shaped channel, the acidic oil phase induces the release of calcium ions from the chelate in the sodium alginate shell; subsequently, divalent Ca... 2+ The two carboxyl groups of alginate combine to form a cross-linked 3D network shell. Subsequently, by raising the temperature from 4°C to 37°C, the matrix gel of the core phase solidifies into a 3D network structure.

[0039] This method not only facilitates the transfer and manipulation of hydrogel microspheres loaded with hiPSCs, but also allows for the loading of different types of cells into the shell and core space of the microspheres, thus enabling the study of interactions between cells or cell-derived organoids. Attached Figure Description

[0040] Figure 1 This is a schematic diagram of the chip in this application;

[0041] Figure 2 The structural diagram of the chip in this application.

[0042] In the diagram, 1 is the first pipe; 2 is the second pipe; 3 is the third pipe; 4 is the fourth pipe; 5 is the fifth pipe; 6 is the sixth pipe; and 7 is the main pipe. Detailed Implementation

[0043] The present application will now be described in detail. While specific embodiments of the present application are shown, it should be understood that the present application can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to enable a more thorough understanding of the present application and to fully convey the scope of the present application to those skilled in the art.

[0044] It should be noted that certain terms are used in the specification and claims to refer to specific components. Those skilled in the art will understand that different terms may be used to refer to the same component. This specification and claims do not distinguish components based on differences in terminology, but rather on differences in function. The terms "comprising" or "including" used throughout the specification and claims are open-ended and should be interpreted as "comprising but not limited to." The following descriptions in the specification are preferred embodiments for carrying out this application; however, these descriptions are for the purpose of understanding the general principles of the specification and are not intended to limit the scope of this application. The scope of protection of this application shall be determined by the appended claims.

[0045] refer to Figure 1 This application provides a droplet microfluidic chip, including a main channel 7 through which liquid flows, and a first channel 1 and a second channel 2; the first channel 1 and the second channel 2 are respectively connected to the main channel 7, and the first channel 1 and the second channel 2 are respectively located on opposite sides of the main channel 7; the same liquid flows in the first channel 1 and the second channel 2.

[0046] A third pipe 3 and / or a fourth pipe 4 are intersecting and connected on the main pipe 7;

[0047] The third pipe 3 and / or the fourth pipe 4 can divide the continuously flowing liquid in the main pipe 7 into droplets.

[0048] The third pipe 3 and / or the fourth pipe 4 are located downstream of the first pipe 1 and the second pipe 2.

[0049] refer to Figure 1 The main pipe 7 contains a continuously flowing liquid. Then, the first pipe 1, the second pipe 2, the third pipe 3 and the fourth pipe 4, which are connected to the main pipe 7 respectively, work together to encapsulate, cut and solidify the liquid in the main pipe 7 by inputting liquid into the main pipe 7.

[0050] In this application, the first pipe 1 and the second pipe 2 are connected to the main pipe 7, allowing the liquid in the first pipe 1 and the second pipe 2 to be transported into the main pipe 7. The first pipe 1 and the second pipe 2 are located on opposite sides of the main pipe 7, thus enabling the liquid in the first pipe 1 and the second pipe 2 to completely encapsulate the liquid in the main pipe 7. Furthermore, when the liquid in the first pipe 1 and the second pipe 2 encapsulates the liquid in the main pipe 7, it can be that the liquid in the first pipe 1 first encapsulates a portion of the liquid in the main pipe 7, and then the remaining portion is encapsulated by the liquid in the second pipe 2. Alternatively, the liquid in the first pipe 1 and the second pipe 2 can simultaneously encapsulate the liquid in the main pipe 7.

[0051] The third pipe 3 and the fourth pipe 4 are also connected to the main pipe 7. When the liquid in the main pipe 7 flows to the position of the third pipe 3 and the fourth pipe 4, the liquid in the third pipe 3 and the fourth pipe 4 also flows into the main pipe 7. Then, when the liquid in the third pipe 3 and the fourth pipe 4 flows into the main pipe 7, it will generate a shearing force on the liquid in the main pipe 7, thereby cutting the liquid in the main pipe 7 into droplets.

[0052] In this application, by connecting the first pipe 1 and the second pipe 2 to the main pipe 7, the liquid in the main pipe 7 can be enveloped by the liquids in the first pipe 1 and the second pipe 2. Then, with the addition and action of the liquids in the third pipe 3 and the fourth pipe 4, the liquid in the main pipe 7 is sheared, thereby realizing the inclusion and cutting of the liquid flowing inside the main pipe 7 into droplets. This ensures that the liquid in the main pipe 7 meets the requirements for subsequent processing.

[0053] The first connection point is formed at the intersection of the first pipe 1 and the main pipe 7, and the second connection point is formed at the intersection of the second pipe 2 and the main pipe 7; preferably, the first connection point and the second connection point coincide.

[0054] In one specific embodiment, the first connection and the second connection are offset from each other, and the first connection is located upstream of the second connection.

[0055] In another specific embodiment, the first connection and the second connection are offset, and the first connection is located downstream of the second connection.

[0056] In one specific embodiment, the first connection point and the second connection point are arranged to overlap.

[0057] The liquid in the first pipe 1 and the second pipe 2 is used to wrap the liquid in the main pipe 7. Therefore, the first connection and the second connection can be staggered so that the liquid in the first pipe 1 and the second pipe 2 can wrap the liquid inside the main pipe 7 on both sides.

[0058] In one specific embodiment, the diameter of the main pipe 7 gradually increases from upstream to downstream.

[0059] Because the liquids inside the first pipe 1 and the second pipe 2 are connected to and flow into the main pipe 7, the amount of liquid in the main pipe 7 increases, causing the diameter of the main pipe 7 to gradually increase. As the liquid flows through the main pipe 7, the flow rate decreases accordingly with the increased diameter. This allows the flow rate of the liquid in the main pipe 7 to be controlled within a reasonable range, making it easier to encapsulate or cut the liquid.

[0060] The diameters of the first pipe 1 and the second pipe 2 are equal or similar.

[0061] Since the first pipe 1 and the second pipe 2 are located on opposite sides of the main pipe 7 and respectively input liquid into the main pipe 7, in order to ensure that the liquid in the main pipe 7 is evenly enveloped by the first pipe 1 and the second pipe 2, their diameters are set to be equal or similar. This allows for a more uniform envelopment of the liquid in the main pipe 7 by the liquid in the first pipe 1 and the second pipe 2, while also achieving a complete containment of the liquid in the main pipe 7.

[0062] The first or second pipe has a diameter of 50 μm to 150 μm.

[0063] Specifically, the diameter of the first pipe or the second pipe is 50 μm, 51 μm, 52 μm, 53 μm, 54 μm, 55 μm, 56 μm, 57 μm, 58 μm, 59 μm, 60 μm, 61 μm, 62 μm, 63 μm, 64 μm, 65 μm, 66 μm, 67 μm, 68 μm, 69 μm, 70 μm, 71 μm, 72 μm, 73 μm, 74 μm, 75 μm, 76 μm, 77 μm, 78 μm, 79 μm, 80 μm, 81 μm, 82 μm, 83 μm, 84 μm, 85 μm, 86 μm, 87 μm, 88 μm, 89 μm, 90 μm, 91 μm, 92 μm, 93 μm, 94μm, 95 μm, 96 μm, 97 μm, 98 μm, 99 μm, 100 μm, 101 μm, 102 μm, 103 μm, 104 μm, 105 μm, 106 μm, 107 μm, 108 μm, 109 μm, 110 μm, 111 μm, 112 μm, 113 μm, 114 μm, 115 μm, 116 μm, 117 μm, 118 μm, 119 μm, 130 μm, 131 μm, 132 μm, 133 μm, 134 μm, 135 μm, 136 μm, 137 μm, 138 μm, 139 μm, 140 μm, 141 μm, 142 μm, 143 μm, 144 μm, 145 μm, 146 μm, 147 μm, 148 μm, 149 μm, 150 μm.

[0064] The main pipe 7 contains a core liquid, the first pipe 1 contains a first liquid, and the second pipe 2 contains a second liquid. Preferably, both the first liquid and the second liquid flow toward the interior of the main pipe 7. More preferably, the core liquid, the first liquid, and the second liquid merge at the first connection and / or the second connection to form a fifth liquid.

[0065] The core liquid flows upstream of the first pipe 1 and the second pipe 2 in the main pipe 7. After passing through the first pipe 1 and the second pipe 2, the core liquid is enveloped by the first liquid and the second liquid, thus forming the fifth liquid.

[0066] In one specific embodiment, the first liquid and the second liquid are the same liquid.

[0067] In another specific embodiment, the first liquid and the second liquid can be different liquids.

[0068] Both the first liquid and the second liquid are liquids that are not easily soluble in the core liquid, so the first liquid and the second liquid can simultaneously encapsulate the core liquid.

[0069] The flow rate of the core liquid in the main pipeline is 1~20 μL / min; preferably, the flow rate of the core liquid in the main pipeline is 2~10 μL / min.

[0070] Specifically, the flow rates of the core fluid in the main pipeline are: 1 μL / min, 2 μL / min, 3 μL / min, 4 μL / min, 5 μL / min, 6 μL / min, 7 μL / min, 8 μL / min, 9 μL / min, 10 μL / min, 11 μL / min, 12 μL / min, 13 μL / min, 14 μL / min, 15 μL / min, 16 μL / min, 17 μL / min, 18 μL / min, 19 μL / min, and 20 μL / min.

[0071] The flow rate of the first liquid in the first pipe is 1~10 μL / min; more preferably, the flow rate of the first liquid in the first pipe is 2~5 μL / min.

[0072] Specifically, the flow rates of the first liquid in the first pipe are 1 μL / min, 2 μL / min, 3 μL / min, 4 μL / min, 5 μL / min, 6 μL / min, 7 μL / min, 8 μL / min, 9 μL / min, and 10 μL / min.

[0073] The flow rate of the second liquid in the second pipe is 1~10 μL / min; more preferably, the flow rate of the second liquid in the second pipe is 2~5 μL / min.

[0074] Specifically, the flow rates of the second liquid in the second pipe are 1 μL / min, 2 μL / min, 3 μL / min, 4 μL / min, 5 μL / min, 6 μL / min, 7 μL / min, 8 μL / min, 9 μL / min, and 10 μL / min.

[0075] When preparing droplet microspheres at different flow rates using the core liquid and the first and second liquids, a relatively large flow rate of the core liquid results in a larger core phase volume and a smaller outer shell phase thickness for the prepared droplet microspheres. Therefore, in this application, the flow rates of the core liquid, as well as the first and second liquids, are controlled within a reasonable range to ensure that the core and outer shell phases of the prepared droplet microspheres have suitable dimensions.

[0076] The fifth liquid flows in the main pipe 7.

[0077] After the core liquid is encapsulated to form the fifth liquid, it continues to flow downstream in the main pipe 7. Then, the third pipe 3 and / or the fourth pipe 4 connected to the main pipe 7 can cut the fifth liquid downstream, so that the fifth liquid can form droplets (i.e. microspheres).

[0078] The flow rate of the first liquid is equal to or similar to that of the second liquid; preferably, the cross-sectional diameter and flow rate of the first liquid and the second liquid are equal to or similar.

[0079] To ensure a more uniform coating of the core liquid by the first and second liquids, their flow diameters are controlled to be equal or similar. This way, when the first and second liquids flow into the main pipe 7, they will diffuse evenly to the outer periphery of the core liquid, uniformly coating it.

[0080] A third pipe 3 and a fourth pipe 4 are intersecting and connected on the main pipe 7;

[0081] Preferably, the third pipe 3 and the fourth pipe 4 are positioned opposite each other on both sides of the main pipe 7.

[0082] In one specific embodiment, the main pipe 7 is connected only to the third pipe 3.

[0083] In one specific embodiment, the main pipe 7 is connected only to the fourth pipe 4.

[0084] In one specific embodiment, the main pipe 7 is connected to a third pipe 3 and a fourth pipe 4.

[0085] Since the third pipe 3 and the fourth pipe 4 are used to cut the fifth liquid in the main pipe 7, the cutting effect can be achieved by only setting either the third pipe 3 or the fourth pipe 4. Preferably, the third pipe 3 and the fourth pipe 4 are set on the main pipe 7 simultaneously, and the third pipe 3 and the fourth pipe 4 are arranged opposite each other on both sides of the main pipe 7. The third pipe 3 and the fourth pipe 4 are used to cut the liquid in the main pipe 7 at the same time, which improves the cutting efficiency. Furthermore, the flow of the liquid in the third pipe 3 and the fourth pipe 4 can be controlled to achieve control over the state such as the interval or size of the droplets.

[0086] The diameter of the third pipe 3 is equal to or similar to the diameter of the fourth pipe 4; preferably, the diameters of both the third pipe 3 and the fourth pipe 4 are less than or equal to the diameter of the main pipe 7.

[0087] The intersection of the third pipe 3 and the main pipe 7 forms a third connection, and the intersection of the fourth pipe 4 and the main pipe 7 forms a fourth connection; preferably, the third connection and the fourth connection coincide.

[0088] A third liquid flows inside the third pipe 3, and a fourth liquid flows inside the fourth pipe 4. The third liquid and the fourth liquid can shear the liquid in the main pipe 7 into droplets.

[0089] The diameter of the third or fourth pipe is 50 μm to 200 μm.

[0090] Specifically, the diameters of the third pipe or the fourth pipe are: 50 μm, 51 μm, 52 μm, 53 μm, 54 μm, 55 μm, 56 μm, 57 μm, 58 μm, 59 μm, 60 μm, 61 μm, 62 μm, 63 μm, 64 μm, 65 μm, 66 μm, 67 μm, 68 μm, 69 μm, 70 μm, 71 μm, 72 μm, 73 μm, 74 μm, 75 μm, 76 μm, 77 μm, 78 μm, 79 μm, 80 μm, 81 μm, 82 μm, 83 μm, 84 μm, 85 μm, 86 μm, 87 μm, 88 μm, 89 μm, 90 μm, 91 μm, 92 μm, 93 μm, 94 μm, 95 μm, 96 μm, 97 μm, 98 μm, 99 μm, 100 μm, 101 μm, 102 μm, 103 μm, 104 μm, 105 μm, 106 μm, 107 μm, 108 μm, 109 μm, 110 μm, 111 μm, 112 μm, 113 μm, 114 μm, 115 μm, 116 μm, 117 μm, 118 μm, 119 μm, 130 μm, 131 μm, 132 μm, 133 μm, 134 μm, 135 μm, 136 μm, 137 μm, 138 μm, 139 μm, 140 μm, 141 μm, 142 μm, 143 μm, 144 μm, 145 μm, 146 μm, 147 μm, 148μm, 149μm, 150μm, 151μm, 152μm, 153μm, 154μm, 155μm, 156μm, 157μm, 158μm, 159μm, 160μm, 161μm, 162μm, 163μm, 164μm, 165μm, 166μm, 167μm, 168μm, 169μm, 170μm, 171μm, 172μm, 173μm, 174μm, 175μm, 176μm, 177μm, 178μm, 179μm, 180μm, 181μm, 182μm, 183μm, 184μm, 185μm, 186μm, 187μm, 188 μm, 189 μm, 190 μm, 191 μm, 192 μm, 193 μm, 194 μm, 195 μm, 196 μm, 197 μm, 198 μm, 199 μm, 200 μm.

[0091] In one specific embodiment, the diameters of the third and fourth pipes are larger than the diameters of the first and second pipes.

[0092] The third pipe 3 and the fourth pipe 4 are formed by introducing liquid into the main pipe 7, which causes the fifth liquid in the main pipe 7 to be cut by the impact of the third and fourth liquids, thus forming droplets.

[0093] To improve the cutting effect of the fifth liquid, the diameters of the third pipe 3 and the fourth pipe 4 are set to be equal or similar, ensuring that the liquids in the third pipe 3 and the fourth pipe 4 flow into the main pipe 7 at equal flow rates and exert an equal cutting effect on the fifth liquid. This reduces the possibility that the fifth liquid in the main pipe 7 might be scattered by the impact of the third or fourth liquid due to differences in the flow rates of the liquids in the third pipe 3 or the fourth pipe 4.

[0094] The distance from the first pipe to the third or fourth pipe is 500 μm to 1500 μm; more preferably, the distance from the first pipe to the third or fourth pipe is 800 μm to 1200 μm.

[0095] Specifically, the distances from the first pipe to the third or fourth pipe are: 500 μm, 550 μm, 600 μm, 650 μm, 700 μm, 750 μm, 800 μm, 850 μm, 900 μm, 950 μm, 1000 μm, 1050 μm, 1100 μm, 1150 μm, 1200 μm, 1250 μm, 1300 μm, 1350 μm, 1400 μm, 1450 μm, and 1500 μm.

[0096] The flow rate of the third liquid in the third pipe is 1~5 μL / min;

[0097] Specifically, the flow rates of the third liquid in the third pipe are: 1 μL / min, 1.5 μL / min, 2 μL / min, 2.5 μL / min, 3 μL / min, 3.5 μL / min, 4 μL / min, 4.5 μL / min, and 5 μL / min.

[0098] The flow rate of the fourth liquid in the fourth pipe is 1~5 μL / min.

[0099] Specifically, the flow rates of the fourth liquid in the fourth pipe are 1 μL / min, 1.5 μL / min, 2 μL / min, 2.5 μL / min, 3 μL / min, 3.5 μL / min, 4 μL / min, 4.5 μL / min, and 5 μL / min.

[0100] A fifth pipe 5 and / or a sixth pipe 6 are connected to the main pipe 7;

[0101] Preferably, a fifth pipe 5 and a sixth pipe 6 are connected in an intersecting state on the main pipe 7;

[0102] Preferably, the fifth pipe 5 and the sixth pipe 6 are arranged opposite each other on both sides of the main pipe 7;

[0103] More preferably, the fifth pipe 5 and the sixth pipe 6 are both located downstream of the third pipe 3 and the fourth pipe 4.

[0104] The sixth liquid flows inside both the fifth pipe 5 and the sixth pipe 6.

[0105] In one specific embodiment, the main pipe 7 is connected only to the fifth pipe 5.

[0106] In one specific embodiment, the main pipe 7 is connected only to the sixth pipe 6.

[0107] In one specific embodiment, the main pipe 7 is simultaneously connected to a fifth pipe 5 and a sixth pipe 6.

[0108] After the fifth liquid is cut into droplets by the third and fourth liquids, a sixth liquid can be placed downstream of the fifth liquid to process the droplets of the fourth liquid, thus subjecting the liquid containing the core liquid to secondary processing by the first and second liquids. This ensures that the droplets of the fifth liquid achieve the desired effect. Simultaneously, to reduce the damaging effect of the sixth liquid on the droplets in the fifth and sixth pipes 5 and 6, the fifth and sixth pipes 5 and 6 are respectively located on opposite sides of the main pipe 7, reducing the impact of the sixth liquid on the liquid.

[0109] The diameters of the fifth and sixth pipes are 200~400 μm.

[0110] Specifically, the diameters of the fifth and sixth pipes are: 200 μm, 201 μm, 202 μm, 203 μm, 204 μm, 205 μm, 206 μm, 207 μm, 208 μm, 209 μm, 210 μm, 211 μm, 212 μm, 213 μm, 214 μm, 215 μm, 216 μm, 217 μm, 218 μm, 219 μm, 230 μm, 231 μm, 232 μm, 233 μm, 234 μm, 235 μm, 236 μm, 237 μm, 238 μm, 239 μm, 240 μm, 241 μm, 242 μm, 243 μm, 244 μm, 245 μm, 246 μm, 247 μm, 248 μm, 249 μm, 250 μm, 251 μm, 252 μm, 253 μm, 254 μm, 255 μm, 256 μm, 257 μm, 258 μm, 259 μm, 260 μm, 261 μm, 262 μm, 263 μm, 264 μm, 265 μm, 266 μm, 267 μm, 268 μm, 269 μm, 270 μm, 271 μm, 272 μm, 273 μm, 274 μm, 275 μm, 276 μm, 277 μm, 278 μm, 279 μm, 280 μm, 281 μm, 282 μm, 283 μm, 284 μm, 285 μm, 286 μm, 287 μm, 288 μm, 289 μm, 290 μm, 291 μm, 292 μm, 293 μm, 294 μm, 295 μm, 296 μm, 297 μm, 298 μm, 299 μm, 300 μm, 301 μm, 302 μm, 303 μm, 304 μm, 305 μm, 306 μm, 307 μm, 308 μm, 309 μm, 310 μm, 311 μm, 312 μm, 313 μm, 314 μm, 315 μm, 316 μm, 317 μm, 318 μm, 319 μm, 320 μm, 321 μm, 322 μm, 323 μm, 324 μm, 325 μm, 326 μm, 327 μm, 328 μm, 329 μm, 330 μm, 331 μm, 332 μm, 333 μm, 334 μm, 335 μm, 336 μm, 337 μm, 338 μm, 339 μm, 340 μm, 341 μm, 342 μm, 343 μm, 344 μm, 345 μm, 346 μm, 347 μm, 348 μm, 349μm, 350 μm, 351 μm, 352 μm, 353 μm, 354 μm, 355 μm, 356 μm, 357 μm, 358 μm, 359 μm, 360 μm, 361 μm, 362 μm, 363 μm, 364 μm, 365 μm, 366 μm, 367 μm, 368 μm, 369 μm, 370 μm, 371 μm, 372 μm, 373 μm, 374 μm, 375 μm, 376 μm, 377 μm, 378 μm, 379 μm, 380 μm, 381 μm, 382 μm, 383 μm, 384 μm, 385 μm, 386 μm, 387 μm, 388 μm, 389 μm, 390 μm, 391 μm, 392 μm, 393 μm, 394 μm, 395 μm, 396 μm, 397 μm, 398 μm, 399 μm, 400 μm.

[0111] In one specific embodiment, the diameters of the fifth and sixth pipes are greater than the diameters of the third and fourth pipes.

[0112] The distance between the third pipe and the fifth or sixth pipe is 500 μm to 1500 μm; more preferably, the distance between the first pipe and the third or fourth pipe is 800 μm to 1200 μm.

[0113] Specifically, the distances from the first pipe to the third or fourth pipe are: 500 μm, 550 μm, 600 μm, 650 μm, 700 μm, 750 μm, 800 μm, 850 μm, 900 μm, 950 μm, 1000 μm, 1050 μm, 1100 μm, 1150 μm, 1200 μm, 1250 μm, 1300 μm, 1350 μm, 1400 μm, 1450 μm, and 1500 μm.

[0114] The flow rate (or flow rate) of the fifth liquid in the fifth pipe is 1~5 μL / min;

[0115] Specifically, the flow rates of the fifth liquid in the fifth pipe are: 1 μL / min, 1.5 μL / min, 2 μL / min, 2.5 μL / min, 3 μL / min, 3.5 μL / min, 4 μL / min, 4.5 μL / min, and 5 μL / min.

[0116] The flow rate (or flow rate) of the sixth liquid in the sixth pipe is 1~5 μL / min;

[0117] Specifically, the flow rates of the sixth liquid in the sixth pipe are: 1 μL / min, 1.5 μL / min, 2 μL / min, 2.5 μL / min, 3 μL / min, 3.5 μL / min, 4 μL / min, 4.5 μL / min, and 5 μL / min.

[0118] The chip also includes a first inlet pipe disposed upstream of the main pipe 7, through which liquid flows into the main pipe 7;

[0119] Preferably, the chip further includes a second inlet channel, which is connected to the first channel 1 and the second channel 2;

[0120] Preferably, the chip further includes a third inlet channel, which is connected to the third channel 3 and the fourth channel 4;

[0121] Preferably, the chip further includes a fourth inlet channel, which is connected to the fifth channel 5 and the sixth channel 6.

[0122] The first, second, third, and fourth inlet pipes are used to introduce liquid into the internal channels of the chip, allowing the core liquid to smoothly enter the main pipe 7. The first liquid can enter the first pipe 1, the second liquid can enter the second pipe 2, the third liquid can enter the third pipe 3, the fourth liquid can enter the fourth pipe 4, and the sixth liquid can enter the fifth pipe 5 and the sixth pipe 6.

[0123] In one specific embodiment, the first liquid and the second liquid are the same liquid, so they can be introduced simultaneously using a second inlet pipe. The third liquid and the fourth liquid are the same liquid, so they can be introduced simultaneously using a third inlet pipe. This further reduces the differences between the first and second liquids and between the third and fourth liquids. It also improves the purity of the manufactured liquid and reduces impurities.

[0124] The diameter of the main pipeline gradually increases along the direction of the first, third, and fifth pipelines.

[0125] Specifically, the diameter of the main pipeline between the first pipeline and the third pipeline is greater than the diameter of the main pipeline located at the position of the first pipeline away from the third pipeline (i.e., the position of the main pipeline upstream of the first pipeline).

[0126] Similarly, the diameter of the main pipe between the third and fifth pipes is greater than the diameter of the main pipe between the first and third pipes. The diameter of the main pipe downstream of the fifth pipe is greater than the diameter of the main pipe between the third and fourth pipes.

[0127] The diameter of the main pipeline at the upstream position of the first pipeline is 200 μm to 250 μm;

[0128] Specifically, the diameter of the main pipeline at the upstream position of the first pipeline is: 200 μm, 201 μm, 202 μm, 203 μm, 204 μm, 205 μm, 206 μm, 207 μm, 208 μm, 209 μm, 210 μm, 211 μm, 212 μm, 213 μm, 214 μm, 215 μm, 216 μm, 217 μm, 218 μm, 219 μm, 230 μm, 231 μm, 232 μm, 233 μm, 234 μm, 235 μm, 236 μm, 237 μm, 238 μm, 239 μm, 240 μm, 241 μm, 242 μm, 243 μm, 244 μm, 245 μm, 246 μm, 247 μm, 248 μm, 249 μm, 250 μm.

[0129] The diameter of the main pipeline between the first and third pipelines is 200 μm ~ 300 μm;

[0130] Specifically, the diameters of the main pipe between the first pipe and the third pipe are: 200 μm, 201 μm, 202 μm, 203 μm, 204 μm, 205 μm, 206 μm, 207 μm, 208 μm, 209 μm, 210 μm, 211 μm, 212 μm, 213 μm, 214 μm, 215 μm, 216 μm, 217 μm, 218 μm, 219 μm, 230 μm, 231 μm, 232 μm, 233 μm, 234 μm, 235 μm, 236 μm, 237 μm, 238 μm, 239 μm, 240 μm, 241 μm, 242 μm, 243 μm, 244 μm, 245 μm, 246 μm, 247 μm, 248 μm, 249 μm, 250 μm, 251 μm, 252 μm, 253 μm, 254 μm, 255 μm, 256 μm, 257 μm, 258 μm, 259 μm, 260 μm, 261 μm, 262 μm, 263 μm, 264 μm, 265 μm, 266 μm, 267 μm, 268 μm, 269 μm, 270 μm, 271 μm, 272 μm, 273 μm, 274 μm, 275 μm, 276 μm, 277 μm, 278 μm, 279 μm, 280 μm, 281 μm, 282 μm, 283 μm, 284 μm, 285 μm, 286 μm, 287 μm, 288 μm, 289 μm, 290 μm, 291 μm, 292 μm, 293 μm, 294 μm, 295 μm, 296 μm, 297 μm, 298 μm, 299 μm, 300 μm.

[0131] The diameter of the main pipe between the third pipe and the fifth pipe is: 250 µm~350 µm.

[0132] Specifically, the diameter of the main pipe between the third pipe and the fifth pipe is 251 μm、252 μm、253 μm、254 μm、255 μm、256 μm、257 μm、258 μm、259 μm、260 μm、261 μm、262 μm、263 μm、264 μm、265 μm、266 μm、267 μm、268 μm、269 μm、270 μm、271 μm、272 μm、273 μm、274 μm、275 μm、276μm、277 μm、278 μm、279 μm、280 μm、281 μm、282 μm、283 μm、284 μm、285 μm、286 μm、287μm、288 μm、289 μm, 290 μm, 291 μm, 292 μm, 293 μm, 294 μm, 295 μm, 296 μm, 297 μm, 298 μm, 299 μm, 300 μm, 301 μm, 302 μm, 303 μm, 304 μm, 305 μm, 306 μm, 307 μm, 308 μm, 309 μm, 310 μm, 311 μm, 312 μm, 313 μm, 314 μm, 315 μm, 316 μm, 317 μm, 318 μm, 319 μm, 320 μm, 321 μm, 322 μm, 323 μm, 324 μm, 325 μm, 326 μm, 327 μm, 328 μm, 329 μm, 330 μm, 331μm, 332 μm, 333 μm, 334 μm, 335 μm, 336 μm, 337 μm, 338 μm, 339 μm, 340 μm, 341 μm, 342 μm, 343 μm, 344 μm, 345 μm, 346 μm, 347 μm, 348 μm, 349 μm, 350 μm.

[0133] The diameter of the main pipe at the lower end of the fifth pipe is 400 µm~600 µm.

[0134] Specifically, the diameter of the main pipeline downstream of the fifth pipeline is: 400 μm, 405 μm, 410 μm, 415 μm, 420 μm, 425 μm, 430 μm, 435 μm, 440 μm, 445 μm, 450 μm, 455 μm, 460 μm, 465 μm, 470 μm, 475 μm, 480 μm, 485 μm, 490 μm, 495 μm, 500 μm, 505 μm, 510 μm, 515 μm, 520 μm, 525 μm, 530 μm, 535 μm, 540 μm, 545 μm, 550 μm, 555 μm, 560 μm, 565 μm, 570 μm, 575 μm, 580 μm, 585 μm, 590 μm. μm, 595 μm, 600 μm.

[0135] In summary, this application provides a droplet microfluidic chip. In use, the core liquid to be encapsulated is input into the main channel 7. Then, the first and second liquids to form the outer shell are delivered to the second and third channels 2 and 3, respectively. At the first connection point, the first and second liquids encapsulate the core liquid, forming a fifth liquid encapsulating the core liquid. The fifth liquid continues to flow in the main channel 7 until it reaches the third and fourth channels 3 and 4. The third and fourth liquids flowing into the third and fourth channels 3 and 4 flow into the main channel 7, creating a shearing effect that cuts the fifth liquid in the main channel 7 into droplet shapes. As the third and fourth liquids continue to flow, the sixth liquid flowing in the fifth and sixth channels 5 and 6 enters the main channel 7, encapsulating the fifth liquid and achieving the desired droplet effect.

[0136] Example

[0137] The main channel, first channel, second channel, third channel, fourth channel, fifth channel, and sixth channel are channels formed within the chip substrate. The first and second channels are coaxial, the third and fourth channels are coaxial, and the fifth and sixth channels are coaxial. The chip substrate dimensions are 3 cm * 4 cm; the substrate thickness is 1~10 mm, and the material used for the chip substrate is PDMS.

[0138] The main channels formed within the chip substrate are circular or horseshoe-shaped channels with a cross-sectional area gradually increasing from 225 to 500 μm. The first and second channels formed within the chip substrate are also circular or horseshoe-shaped channels with a cross-sectional diameter of 100 μm. The third and fourth channels formed within the chip substrate are also circular or horseshoe-shaped channels with a cross-sectional diameter of 125 μm. The fifth and sixth channels formed within the chip substrate are also circular or horseshoe-shaped channels with a cross-sectional area of ​​280 μm.

[0139] The flow rate of the core liquid in the main pipeline is 4~10 μL / min; the flow rate of the first and second liquids is 4 μL / min; the flow rate of the third and fourth liquids is 2 μL / min; and the flow rate of the fifth and sixth liquids is 2 μL / min.

[0140] The main pipeline is connected to the first, second, third, fourth, fifth, and sixth pipelines respectively. The first and second pipelines are located upstream of the third and fourth pipelines, and the third and fourth pipelines are located upstream of the fifth and sixth pipelines. The distance between the first and second pipelines and the third and fourth pipelines is 950 μm. The distance between the third and fourth pipelines and the fifth and sixth pipelines is 1050 μm.

[0141] The diameter of the main pipeline upstream of the first pipeline is 225 μm; the diameter of the main pipeline between the first and third pipelines is 250 μm; the diameter of the main pipeline between the third and fifth pipelines is 300 μm; and the diameter of the main pipeline downstream of the fifth pipeline is 500 μm.

[0142] The first inlet pipe is connected to peristaltic pump A, the second inlet pipe is connected to peristaltic pump B, the third inlet pipe is connected to peristaltic pump C, and the fourth inlet pipe is connected to peristaltic pump D. Furthermore, peristaltic pumps A through D are each connected to their respective liquid solutions.

[0143] Peristaltic pump A is connected to a tank containing matrix adhesive and a tank containing hiPSCs.

[0144] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A droplet microfluidic chip, characterized by, The chip comprises a main pipe in which liquid flows, and a first pipe and a second pipe; the first pipe and the second pipe are respectively communicated with the main pipe, and the first pipe and the second pipe are respectively located at opposite sides of the main pipe; the first pipe and the second pipe flow with the same liquid; A third pipe and / or a fourth pipe are communicated with the main pipe in an intersecting manner; The third pipe and / or the fourth pipe can divide the continuously flowing liquid in the main pipe into liquid drops; The third pipe and / or the fourth pipe are arranged at a downstream position of the first pipe and the second pipe; A fifth pipe and a sixth pipe are communicated with the main pipe in an intersecting manner.

2. The chip according to claim 1, characterized in that, The first pipe and the main pipe form a first connection at the intersection, and the second pipe and the main pipe form a second connection at the intersection.

3. The chip of claim 2, wherein, The first connection and the second connection coincide.

4. The chip of claim 1, wherein The diameter of the main pipe gradually increases from an upstream to a downstream direction.

5. The chip of claim 1, wherein, The diameters of the first pipe and the second pipe are equal or similar.

6. The chip of claim 2, wherein, The main pipe flows with core liquid, the first pipe flows with first liquid, and the second pipe flows with second liquid.

7. The chip of claim 6, wherein The first liquid and the second liquid both flow towards the inside of the main pipe.

8. The chip of claim 6, wherein, The core liquid, the first liquid and the second liquid converge to form fifth liquid at the first connection and / or the second connection.

9. The chip of claim 8, wherein, The fifth liquid flows in the main pipe.

10. The chip of claim 6, wherein, The flow rate of the first liquid is equal or similar to that of the second liquid.

11. The chip of claim 10, wherein, The cross-sectional diameters and flow rates of the first liquid and the second liquid are equal or similar.

12. The chip according to any one of claims 1 to 11, wherein A third pipe and a fourth pipe are communicated with the main pipe in an intersecting manner.

13. The chip of claim 12, wherein, The third pipe and the fourth pipe are oppositely arranged at two sides of the main pipe.

14. The chip of claim 12, wherein, The diameter of the third pipe is equal or similar to that of the fourth pipe.

15. The chip of claim 12, wherein, The diameters of the third pipe and the fourth pipe are equal to or less than that of the main pipe.

16. The chip of claim 12, wherein, The third pipe and the main pipe form a third connection at the intersection, and the fourth pipe and the main pipe form a fourth connection at the intersection.

17. The chip of claim 16, wherein, The third connection and the fourth connection coincide.

18. The chip of claim 12, wherein, The third pipe flows with third liquid, and the fourth pipe flows with fourth liquid, and the third liquid and the fourth liquid can shear the liquid in the main pipe into liquid drops.

19. The chip of claim 1, wherein, The fifth pipe and the sixth pipe are oppositely arranged at two sides of the main pipe.

20. The chip of claim 19, wherein, The fifth pipe and the sixth pipe are both located at a downstream position of the third pipe and the fourth pipe.

21. The chip of claim 19, wherein, The fifth pipe and the sixth pipe both flow with sixth liquid.

22. The chip of claim 1, wherein, The chip further comprises a first introduction pipe arranged at an upstream of the main pipe, through which liquid flows into the main pipe.

23. The chip of claim 22, wherein, The chip further comprises a second introduction pipe, which is communicated with the first pipe and the second pipe.

24. The chip of claim 23, wherein, The chip further comprises a third introduction pipe, which is communicated with the third pipe and the fourth pipe.

25. The chip of claim 24, wherein, The chip further comprises a fourth introduction conduit, which communicates with the fifth conduit and the sixth conduit. The chip further comprises a fourth introduction conduit, which communicates with the fifth conduit and the sixth conduit.

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