Micro-fluidic chip runner and single-cell paired micro-fluidic chip

Abstract

The utility model discloses a micro-fluidic chip flow channel and a single-cell paired micro-fluidic chip, the micro-fluidic chip flow channel comprises a microsphere flow channel and at least one separation device, the microsphere flow channel is provided with a microsphere inlet and a microsphere outlet, the separation device is arranged in the microsphere flow channel, microspheres flow through the separation device in the process of flowing from the microsphere inlet to the microsphere outlet, and the microspheres flow through the single-cell paired micro-fluidic chip. The separation device is used for separating the microspheres, so that the microspheres which are relatively close to each other can be separated, gaps are formed among the microspheres, the microspheres can enter the liquid drop singly, and the phenomenon of empty package or multiple package of the liquid drop is avoided. And the separation device separates the microspheres through physical action, so that the stability is good, and the separation effect is good.

Description

Technical Field

[0001] This application relates to the field of microfluidic chip technology, and in particular to microfluidic chip channels and single-cell paired microfluidic chips. Background Technology

[0002] The cross-shaped structure is the most common channel structure in commercial applications of microfluidic single-cell sequencing, allowing microspheres and cells to be encapsulated one-to-one within droplets. High single-encapsulation rates can be achieved by adjusting the microsphere entry rate and droplet formation rate. During this encapsulation process, the hydrogel enters the channel at nearly 100% volume fraction due to its elasticity and deformability. However, fluctuations in input pressure, systematic errors, or differences in hydrogel size can lead to empty or multiple encapsulations of microspheres when they enter the droplet. Utility Model Content

[0003] This application aims to address at least one of the technical problems existing in the prior art. To this end, this application proposes a microfluidic chip channel that can separate microspheres, allowing individual microspheres to enter the droplet, thereby improving the phenomenon of empty or multiple microspheres in the droplet.

[0004] The microfluidic chip flow channel described in the first aspect embodiment of this application includes:

[0005] A microsphere flow channel is provided with a microsphere inlet and a microsphere outlet, and multiple microspheres flow from the microsphere inlet to the microsphere outlet;

[0006] At least one separating device is disposed within the microsphere channel, the separating device being used to separate the microspheres so that gaps appear between the microspheres.

[0007] The microfluidic chip channel according to the first aspect of this application has at least the following beneficial effects: The microfluidic chip channel of this application includes a microsphere channel and at least one separating device. The microsphere channel is provided with a microsphere inlet and a microsphere outlet. The separating device is disposed within the microsphere channel. As the microspheres flow from the microsphere inlet to the microsphere outlet, they pass through the separating device. The separating device separates the microspheres, separating microspheres that are close together, creating gaps between each microsphere. This allows individual microspheres to enter the droplet, avoiding empty or multiple droplet inclusions. The separating device separates the microspheres through physical action, resulting in good stability and excellent separation effect.

[0008] According to some embodiments of this application, the microsphere channel is provided with a shaft for mounting the separating device, and the separating device is rotatably mounted on the shaft.

[0009] According to some embodiments of this application, two separating devices are provided, and the two separating devices are disposed on both sides of the microsphere channel.

[0010] According to some embodiments of this application, the separating device includes a plurality of separating teeth, which are arranged circumferentially around the shaft, and the angles between each separating tooth are equal.

[0011] According to some embodiments of this application, the separating device includes two separating teeth, which are disposed opposite to each other in the radial direction of the shaft.

[0012] According to some embodiments of this application, the separating device is configured as a pendulum rod capable of reciprocating within the microsphere channel, with one end of the pendulum rod disposed on the shaft.

[0013] According to some embodiments of this application, at least two separating devices are provided, and the at least two separating devices are arranged sequentially in the direction from the microsphere inlet to the microsphere outlet.

[0014] According to some embodiments of this application, the microsphere channel is provided with a receiving cavity, and the separating device is disposed within the receiving cavity.

[0015] According to some embodiments of this application, the width of the microsphere channel is 0-999 μm.

[0016] The single-cell paired microfluidic chip described in the second aspect of this application includes the microfluidic chip channel of the first aspect embodiment.

[0017] The single-cell paired microfluidic chip according to the second aspect of this application has at least the following beneficial effects: The single-cell paired microfluidic chip of this application can separate microspheres by using microfluidic chip channels, separating microspheres that are close together, so that there are gaps between each microsphere, and microspheres can enter the droplet individually, avoiding the phenomenon of empty droplet or multiple droplet packs, and ensuring the success rate of single-cell sequencing.

[0018] According to some embodiments of this application, the single-cell paired microfluidic chip includes a dispersed phase channel, a continuous phase channel, and a converging channel. The dispersed phase channel is connected to the microsphere channel, the continuous phase channel is connected to the microsphere channel, and the converging channel is arranged on the same axis as the microsphere channel.

[0019] According to some embodiments of this application, the dispersed phase channel is configured as a cell channel, the continuous phase channel is configured as an oil channel, and the cell channel and the oil channel are arranged sequentially in the direction from the microsphere inlet to the microsphere outlet.

[0020] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0021] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0022] Figure 1 This is a schematic diagram of a structure in the prior art where multiple packages occur;

[0023] Figure 2 This is a schematic diagram of the structure of a microfluidic chip channel according to an embodiment of this application;

[0024] Figure 3 yes Figure 2 The diagram shows a top view of the microsphere flow channel in the microfluidic chip.

[0025] Figure 4 yes Figure 2 The diagram shows a side view of the microsphere flow channel in the microfluidic chip.

[0026] Figure 5 This is a schematic diagram of the structure of a microfluidic chip channel according to another embodiment of this application;

[0027] Figure 6 This is a schematic diagram of the structure of a microfluidic chip channel according to another embodiment of this application;

[0028] Figure 7 This is a schematic diagram of the structure of a microfluidic chip channel according to another embodiment of this application;

[0029] Figure 8 This is a schematic diagram of the structure of a microfluidic chip channel according to another embodiment of this application;

[0030] Figure 9 This is a schematic diagram of the structure of a microfluidic chip channel according to another embodiment of this application;

[0031] Figure 10 This is a schematic diagram of the structure of a single-cell paired microfluidic chip according to an embodiment of this application.

[0032] Figure label:

[0033] Microsphere channel 100, microsphere 110, shaft 120, microsphere inlet 121, microsphere outlet 122, receiving cavity 130;

[0034] Cell flow channel 200, cell 210;

[0035] Oil flow channel 300;

[0036] Flow channel 400;

[0037] Separating device 500, separating teeth 510, swing rod 520. Detailed Implementation

[0038] The embodiments of this application are described in detail below. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this application, and should not be construed as limiting this application.

[0039] In the description of this application, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, inside, outside, etc., are based on the orientation or positional relationship shown in the accompanying drawings and are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application.

[0040] In the description of this application, "several" means one or more, "more than" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0041] In the description of this application, unless otherwise expressly defined, terms such as setting, installing, connecting, assembling, and cooperating should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this application in conjunction with the specific content of the technical solution.

[0042] Reference Figure 1 In related technologies, fluctuations in input pressure, system errors, or differences in hydrogel size can cause empty or multiple inclusions when microspheres enter droplets.

[0043] The following reference Figures 2 to 9 This application describes the microfluidic chip channel of an embodiment.

[0044] The microfluidic chip channel in this embodiment of the application refers to... Figures 2 to 9The system includes a microsphere channel 100 and at least one separator 500. The microsphere channel 100 is provided with a microsphere inlet 121 and a microsphere outlet 122. Multiple microspheres 110 enter the microsphere channel 100 through the microsphere inlet 121 and flow out through the microsphere outlet 122. In related technologies, the phenomenon of empty or multiple microspheres 110 is avoided by adjusting the entry rate of the microspheres 110. However, due to fluctuations in input pressure, system errors, or differences in hydrogel size, multiple microspheres 110 may still enter the microsphere channel 100 together with a relatively close spacing. A separator 500 is disposed within the microsphere channel 100. As the microspheres 110 flow from the microsphere inlet 121 to the microsphere outlet 122, they pass through the separator 500. The separator 500 separates the microspheres 110, separating closely spaced microspheres and creating gaps between them. This allows individual microspheres 110 to enter the droplet, preventing empty or multiple droplet inclusions. The separator 500 separates the microspheres 110 through physical action, resulting in good stability and excellent separation effect.

[0045] It should be noted that in some embodiments, the microsphere channel 100 is used to transport hydrogel microspheres. In other embodiments, the microsphere channel 100 can also be used to transport other microspheres such as protein microspheres and composite microspheres.

[0046] Reference Figure 2 In some embodiments, a shaft 120 for mounting a separator 500 is provided on the microsphere channel 100, and the separator 500 is rotatably mounted on the shaft 120. Specifically, the separator 500 is provided on the microsphere channel 100, and the separator 500 is rotatably mounted on the shaft 120. When the microspheres 110 flow through the separator 500, they can drive the separator 500 to rotate. During rotation, the rotatable separator 500 can impede the movement of the next adjacent microsphere 110, creating a gap between adjacent microspheres 110, thus separating the microspheres 110 and allowing each microsphere 110 to enter the droplet individually, improving the single-packing rate. It should be noted that, since the microspheres 110 are arranged in a row and move unidirectionally within the microsphere flow channel 100, the microsphere 110 that detaches from the separating device 500 first will travel a greater distance than its adjacent, obstructed microsphere 110, thus creating a gap between adjacent microspheres 110. In some embodiments, the shaft 120 and the microsphere flow channel 100 are designed as a single unit. In other embodiments, the shaft 120 and the microsphere flow channel 100 are designed as separate units.

[0047] Reference Figure 2In some embodiments, two separators 500 are provided, and the two separators 500 are arranged on both sides of the microsphere channel 100. The length direction of the microsphere channel 100 is defined as the direction from the microsphere inlet 121 to the microsphere outlet 122 (refer to...). Figure 2 , Figure 2 The X-axis direction is the length direction, and the width direction is set perpendicular to the length direction (refer to...). Figure 2 , Figure 2 (The Y-axis direction is the width direction). Two separating devices 500 are arranged on both sides of the microsphere channel 100 in the width direction. During the hydrogel delivery process, the two separating devices 500 separate the microspheres 110 from both sides in the width direction, resulting in better separation of the microspheres 110. It should be noted that in some embodiments, the two separating devices 500 are arranged opposite to each other in the width direction, and the two oppositely arranged separating devices 500 can achieve synchronous separation of the microspheres 110. It can be understood that the separation of the microspheres 110 by the two separating devices 500 can be performed synchronously or asynchronously. Refer to Figure 7 In some embodiments, the two separating devices 500 move synchronously under the action of the microsphere 110, at which point the two separating devices 500 simultaneously impede the movement of one microsphere 110. (Refer to...) Figure 8 In some other embodiments, the two separating devices 500 move asynchronously, with one separating device 500 hindering the movement of the microsphere 110 and the other separating device 500 hindering the movement of the adjacent microsphere 110, thus achieving the separation of the microsphere 110.

[0048] Reference Figure 2 and Figure 5 In some embodiments, the separating device 500 includes a plurality of separating teeth 510, which are arranged circumferentially around the shaft 120, with equal angular spacing between each separating tooth 510. The separating device 500 is rotatably mounted on the shaft 120. When the microsphere 110 flows through the separating device 500, it drives the separating teeth 510 of the separating device 500 to rotate. Since there are multiple separating teeth 510, when a microsphere 110 abuts against the first separating tooth 510 and drives the separating device 500 to rotate, the other rotating separating teeth 510 will obstruct the next microsphere 110, creating a gap between adjacent microspheres 110, thus separating the microspheres 110 and allowing each microsphere 110 to enter the droplet individually, improving the single-packing rate. Multiple separating teeth 510 are arranged circumferentially around the shaft 120, with equal angular spacing between each separating tooth 510. These teeth 510 can rotate cyclically around the shaft 120 under the influence of multiple microspheres 110, continuously separating the microspheres 110. In some embodiments, the number of separating teeth 510 can be three, four, five, or more, all of which can achieve the function of separating the microspheres 110.

[0049] Reference Figure 7 and Figure 8 In some embodiments, the separating device 500 includes two separating teeth 510, which are arranged opposite each other in the radial direction of the shaft 120. The two separating teeth 510 are rotatable about the shaft 120. A microsphere 110 abuts against one end of a separating tooth 510 and drives the tooth 510 to rotate. The other end of the rotating separating tooth 510 obstructs the next microsphere 110, creating a gap between adjacent microspheres 110, thus separating the microspheres 110 and allowing each microsphere 110 to enter the droplet individually, improving the single-packing rate. In some embodiments, the two separating teeth 510 are designed separately. In other embodiments, the two separating teeth 510 are designed as a single unit.

[0050] Reference Figure 9 It is understood that in some other embodiments, the separating device 500 can also be configured as a swing arm 520 that can reciprocate within the microsphere channel 100. One end of the swing arm 520 is mounted on a shaft. When the microsphere 110 passes by, the swing arm 520 obstructs the microsphere 110. The microsphere 110 abuts against and drives the swing arm 520 to move. After the microsphere 110 passes the position of the swing arm 520, when the swing arm 520 returns to its lowest point, it blocks the subsequent microsphere 110, creating a gap between two adjacent microspheres 110, thus achieving the separation of the microspheres 110. It is understood that the reciprocating motion of the swing arm 520 within the microsphere channel 100 can be a reciprocating swing between high and low points, or a reciprocating rotation within the microsphere channel 100.

[0051] Reference Figure 6 In some embodiments, two separators 500 are provided, and the two separators 500 are arranged sequentially in the direction from the microsphere inlet 121 to the microsphere outlet 122. The direction from the microsphere inlet 121 to the microsphere outlet 122 is defined as the length direction of the microsphere flow channel 100 (refer to...). Figure 6 , Figure 6 (The X-axis direction is the length direction). Two separating devices 500 are sequentially arranged along the length of the microsphere channel 100. When multiple microspheres 110 move from the glue inlet to the glue outlet, the microspheres 110 pass through the two separating devices 500 in sequence. The two sequentially arranged separating devices 500 can perform secondary separation of the microspheres 110, preventing adjacent microspheres 110 from not being separated in the first separation, ensuring the separation effect and improving the single-pack yield. It is understood that in some other embodiments, the number of separating devices 500 can also be set to multiple, and multiple separating devices 500 can separate the microspheres 110 multiple times, resulting in a better separation effect.

[0052] Reference Figure 5 In some embodiments, the microsphere channel 100 is provided with a receiving cavity 130, and the separating device 500 is disposed within the receiving cavity 130. The direction from the microsphere inlet 121 to the microsphere outlet 122 is defined as the length direction of the microsphere channel 100 (refer to...). Figure 5 , Figure 5 The X-axis direction is the length direction, and the width direction is set perpendicular to the length direction (refer to...). Figure 5 , Figure 5 (The Y-axis direction is the width direction). The receiving cavity 130 is located in the width direction of the microsphere flow channel 100. The receiving cavity 130 is used to accommodate the separating device 500, providing space for the separating device 500 to move within the microsphere flow channel 100 and separate the microspheres 110 flowing in the microsphere flow channel 100. It can be understood that, referring to... Figure 2 , Figure 3 and Figure 4 In some embodiments, two separators 500 are provided, and two corresponding receiving cavities 130 are provided. The two receiving cavities 130 are arranged opposite each other in the width direction of the microsphere channel 100 to accommodate the two separators 500.

[0053] In some embodiments, the width of the microsphere channel 100 is 0-999 μm. The direction from the microsphere inlet 121 to the microsphere outlet 122 is defined as the length direction of the microsphere channel 100 (refer to...). Figure 5 , Figure 5 The X-axis direction is the length direction, and the width direction is set perpendicular to the length direction (refer to...). Figure 5 , Figure 5 The width of the microsphere channel 100 (with the Y-axis as the width direction) is 0-999μm, which can transport micron-sized microspheres 110.

[0054] Reference Figure 10 The single-cell paired microfluidic chip of the second aspect of this application includes the microfluidic chip channel of the first aspect embodiment. The single-cell paired microfluidic chip of this application, by using the microfluidic chip channel, can separate microspheres 110, separating closely spaced microspheres 110, creating gaps between each microsphere 110. This allows each microsphere 110 to enter the droplet individually, avoiding empty or multiple droplet packets and ensuring the success rate of single-cell sequencing.

[0055] In some embodiments, the single-cell paired microfluidic chip includes a dispersed phase channel, a continuous phase channel, and a collecting channel 400. The dispersed phase channel is connected to the microsphere channel 100, and the continuous phase channel is also connected to the microsphere channel 100. The collecting channel 400 and the microsphere channel 100 are arranged on the same axis. A dispersed phase fluid carrying cells 210 is transported in the dispersed phase channel. By adjusting the size of the dispersed phase channel and the flow rate of the fluid to suitable parameters, cells 210 can be inserted into droplets one-to-one. Because the droplets are independent of each other, each droplet can provide a relatively closed microreaction environment, and there is no cross-contamination between the cells 210. Therefore, the droplets can be used as an ideal microreactor. In addition to cells 210, droplets can also be used for target substances such as proteins and nucleic acids. The dispersed phase fluid and the continuous phase fluid are immiscible or partially miscible. In some embodiments, a single-cell pairing microfluidic chip is used to construct single-cell droplets with microspheres 110. The dispersed phase channel is configured as a cell channel 200, and the continuous phase channel is configured as an oil channel 300. The cell channel 200 and the oil channel 300 are arranged sequentially in the direction from the microsphere inlet 121 to the microsphere outlet 122. The cell channel 200 of the single-cell pairing microfluidic chip carries cells 210, and the microsphere channel 100 carries microspheres 110 separated by the separator 500. The continuous phase channel is the oil channel 300. The cell channel 200 and the oil channel 300 are arranged sequentially in the direction from the microsphere inlet 121 to the microsphere outlet 122. When cells 210 and microspheres 110 converge, they combine together, with one cell 210 matching one microsphere 110. The matched cell 210-microsphere 110 combination continues to move through the oil channel 300, forming a cell 210-microsphere 110 droplet wrapped in the continuous phase, which enters the converging channel 400.

[0056] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of this application.

Claims

1. A microfluidic chip flow channel, characterized in that, include: A microsphere flow channel is provided with a microsphere inlet and a microsphere outlet, and multiple microspheres flow from the microsphere inlet to the microsphere outlet; At least one separating device is disposed within the microsphere channel, the separating device being used to separate the microspheres so that gaps appear between the microspheres.

2. The microfluidic chip channel according to claim 1, characterized in that, The microsphere channel is provided with a shaft for mounting the separator, and the separator is rotatably mounted on the shaft.

3. The microfluidic chip channel according to claim 1, characterized in that, Two separation devices are provided, and the two separation devices are located on both sides of the microsphere channel.

4. The microfluidic chip channel according to claim 2, characterized in that, The separating device includes a plurality of separating teeth, which are arranged circumferentially around the shaft, and the angles between each separating tooth are equal.

5. The microfluidic chip channel according to claim 2, characterized in that, The separating device includes two separating teeth, which are arranged opposite each other in the radial direction of the shaft.

6. The microfluidic chip channel according to claim 2, characterized in that, The separating device is configured as a pendulum rod that can reciprocate within the microsphere channel, with one end of the pendulum rod mounted on the shaft.

7. The microfluidic chip channel according to claim 1, characterized in that, At least two separation devices are provided, and the at least two separation devices are arranged sequentially in the direction from the microsphere inlet to the microsphere outlet.

8. The microfluidic chip channel according to claim 1, characterized in that, The microsphere channel is provided with a receiving cavity, and the separating device is disposed within the receiving cavity.

9. The microfluidic chip channel according to claim 1, characterized in that, The width of the microsphere channel is 0-999 μm.

10. A single-cell paired microfluidic chip, characterized in that, Includes the microfluidic chip channel as described in any one of claims 1 to 9.

11. The single-cell paired microfluidic chip according to claim 10, characterized in that, The single-cell paired microfluidic chip includes a dispersed phase channel, a continuous phase channel, and a converging channel. The dispersed phase channel is connected to the microsphere channel, the continuous phase channel is connected to the microsphere channel, and the converging channel is arranged on the same axis as the microsphere channel.

12. The single-cell paired microfluidic chip according to claim 11, characterized in that, The dispersed phase channel is configured as a cell channel, and the continuous phase channel is configured as an oil channel. The cell channel and the oil channel are arranged sequentially in the direction from the microsphere inlet to the microsphere outlet.

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