An acoustofluidic chip based on an asymmetric multi-cusp structure

Abstract

The invention discloses an acoustic fluidic chip based on an asymmetric multi-pointed angle structure, comprising an open acoustic fluidic chip or a closed acoustic fluidic chip based on gapless acoustic fluidic units or gapped acoustic fluidic units; the two types of units have right-angled triangle-shaped sharp corners and right-angled edges, with a cell capture flow channel in the middle; the cell capture flow channel is connected to the main flow channel above, and the left side of the right-angled triangle sharp corner and the right side of the right-angled edge are the connecting parts between the units; the gapless right-angled corner is the same thickness as the right-angled edge and the connecting part, and the gapped right-angled corner is thinner than the right-angled edge and the connecting part; an array of asymmetric multi-pointed angle acoustic fluidic units with or without gaps and a pressure-guided channel form an open acoustic fluidic chip, and an array of asymmetric multi-pointed angle acoustic fluidic units with or without gaps symmetrically arranged along the center line form a closed acoustic fluidic chip with the pressure-guided channel, thereby realizing three-dimensional rotation, arrayed capture and manipulation of micro-scale particles such as cells.

Description

Technical Field

[0001] The present invention belongs to the field of microfluidics technology, and in particular relates to an acoustofluidic chip based on an asymmetric multi-cusp structure. Background Art

[0002] Biological cells regulate a range of fundamental life activities, including growth, proliferation, differentiation, and apoptosis. Modern medicine has shown that the onset and progression of many human diseases are closely linked to changes in cellular physiological and physical properties. Research has found that the mechanical and physical properties of cells exhibit significant anisotropy, meaning that cell stiffness, viscosity, and other properties measured at different test locations vary significantly. Other studies have shown that structural instability and functional defects in cells are directly related to the three-dimensional morphology of internal components, such as the nucleus, mitochondria, and microtubules. By varying the spatial orientation of cells and securing target cells in an array, micro-nano force sensors can precisely control the three-dimensional test locations for cell mechanical properties. Microscopic optical imaging systems can sample images of organelles at multiple rotational angles and reconstruct these images in three dimensions. The resulting analysis of cellular physiological and mechanical properties has important theoretical and practical implications for rapid disease detection and diagnosis, drug screening, and evaluation.

[0003] The complex biological culture environment and the submicron scale of manipulated objects often make efficient, high-throughput, and low-invasive cell rotation difficult to achieve. Some current studies utilize the load force generated by the contact between microneedles and target cells to rotate cells. However, this method can only treat one cell at a time, resulting in low efficiency. Furthermore, the contact force between the microneedle and the cell is difficult to measure and control, which can easily cause irreversible damage to the cells, ultimately leading to cell breakage or even death. Some scholars have also carried out cell movement operations based on microfluidic technology in existing research. However, in order to generate fluid vortex streamlines for cell rotation, most current studies design complex bypass channels around the main channel to generate bypass jets (HARADA H, KANEKOM, ITO H. Rotational manipulation of a microscopic object inside a microfluidicchannel Rotational manipulation of a microscopic object inside a microfluidicchannel[J / OL]. 2020, 054106(May). https: / / doi.org / 10.1063 / 5.0013309). However, this method is difficult to achieve closed fluid vortexes, so the cell rotation rate of this type of device is low; at the same time, due to the need for complex bypass channels, the structural design of this type of device is complex, and chip processing and preparation are difficult.

[0004] Using an external sound source with controllable frequency and amplitude, acoustofluidics can convert external sound source vibrations into closed acoustic flows within microchannels, which can drive cells at high speeds. Traditional acoustofluidics chips (NAMA N, HUANG PH, HUANG TJ et al. Investigation of acoustic streaming patterns around oscillating sharp edges [J / OL]. Lab on a Chip, 2014, 14(15): 2824-2836.

[0005] https: / / doi.org / 10.1039 / c4lc00191e; OZCELIK A, NAMA N, HUANG PH, et al. Acoustofluidic Rotational Manipulation of Cells and Organisms UsingOscillating Solid Structures[J / OL]. Small, 2016, 12(37):5120-5125.

[0006] https: / / onlinelibrary.wiley.com / doi / 10.1002 / smll.201601760) Based on the sharp corners of an equilateral triangle, two symmetrical closed streamlines are generated around the sharp corners. Cells and other particles flowing near this structure are attracted and produce planar rotational motion. However, this type of chip cannot achieve three-dimensional rotation of cells, making it difficult to achieve three-dimensional imaging of cells and internal components. This type of chip often requires a large-amplitude external sound source, and the rate of cell rotation driven by a low-energy sound source needs to be further improved. Furthermore, this type of device cannot capture target cells in an array, and it is even more difficult to maintain cell stability during subsequent cell mechanical property testing and other micromanipulation processes that require fixation. Therefore, there is an urgent need to design a cell micromanipulation acoustofluidic chip that integrates three-dimensional rotation, array capture, and fixation. Summary of the Invention

[0007] In order to overcome the shortcomings of the above-mentioned prior art, the purpose of the present invention is to propose an acoustofluidic chip based on an asymmetric multi-pointed structure, which can be used for three-dimensional rotation, arrayed capture and fixed manipulation of micro-scale particles such as cells.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] An acoustofluidic chip based on an asymmetric multi-cusp structure, including an open acoustofluidic chip or a closed acoustofluidic chip based on a gapless acoustofluidic unit 10 or a gapped acoustofluidic unit 20;

[0010] The gapless acoustic fluidic unit 10 includes a first right-angled triangle-shaped corner 101 and a first right-angled edge 102, with a first cell capture channel 103 located between the two. The first cell capture channel 103 has a first capture channel inlet 1031 at its top, a first sidewall 1032 below the left-hand corner 101, a second sidewall 1033 and a first capture channel outlet 1034 below the right-hand right-angled edge, and a first capture channel bottom edge 1035 at its bottom. The first cell capture channel 103 is connected to the first main channel 104 above. The left side of the first right-angled triangle corner 101 and the right side of the first right-angled edge 102 form a first inter-unit connection 105. The two right-angled sides of the first right-angled triangle corner 101 are located near the first main channel 104 and the first connection 105, respectively. The first right-angled triangle corner 101 has the same thickness as the first right-angled edge 102 and the first connection 105.

[0011] The first capture channel outlet 1034 in the gapless acoustic fluidic unit 10 is arranged as one or more as needed, and is arranged below the first side wall 1032 of the first cell capture channel 103, below the second side wall 1033 and at the bottom edge 1035 of the first capture channel.

[0012] The gapless acoustic fluidic unit 10 is prepared into a microfluidic chip using a reverse molding process. When the material is poured into the unit pattern arranged in an array, the pattern will be covered, and an upper wall of a certain thickness will be formed above the unit. This thickness is used to lift the chip wafer with the unit pattern characteristics from the mold and divide it into independent chip devices.

[0013] The first open acoustic fluidic chip 100 based on the gapless acoustic fluidic unit 10: includes the gapless acoustic fluidic units 10 distributed in an array, the first capture flow channel outlet 1034 of each gapless acoustic fluidic unit 10 is connected through the first pressure guide channel 1001, and accumulated through the first pressure outlet 1002; negative pressure is applied at the first pressure outlet 1002, which can generate negative pressure in the first cell capture flow channel 103 of each gapless acoustic fluidic unit 10 through the first pressure guide channel 1001, thereby fixing the captured cells; the outside of the gapless acoustic fluidic unit 10 is the first main flow channel 104, and above the first main flow channel 104 there is a first main flow channel upper Wall 1041, the outside of the first main channel 104 is the first open fluid environment 1010, and the target cells are placed in the first open fluid environment 1010 using a pipetting device. When the sound waves are transmitted to the sharp corner 101 of the first right-angled triangle and the first cell capture channel 103, high-speed vortices can be generated, causing cells and other micro-scale particles to produce three-dimensional rotational motion. When they are rotated to a suitable angle, the capture function of the chip can be used to fix the target cells in the first cell capture channel 103. The external operating tool moves from the first open fluid environment 1010 to the gap below the upper wall 1041 of the first main channel to contact the captured and fixed cells and carry out subsequent operations.

[0014] The first closed acoustic fluidic chip 200 based on the gapless acoustic fluidic unit 10 includes an array of gapless acoustic fluidic units 10 symmetrically distributed along the midline, wherein the first cell capture flow channel 103 of each gapless acoustic fluidic unit 10 is connected through a second pressure guide channel 2001 and accumulated through a second pressure outlet 2002; negative pressure is applied at the second pressure outlet 2002, which can generate negative pressure in the first cell capture flow channel 103 of each gapless acoustic fluidic unit 10 through the second pressure guide channel 2001, thereby fixing the captured cells; two gapless acoustic fluidic units 10 symmetrically distributed above and below The upper wall 1041 of the first main flow channel outside the array of acoustic fluidic control units 10 forms a first closed operating environment 2010. The target cells are input into the first closed operating environment 2010 through the first material input port 2004 via the first input flow channel 2003. When the sound waves are transmitted to the sharp corner 101 of the first right-angled triangle and the first cell capture flow channel 103, high-speed vortices are generated, causing micro-scale particles such as cells to produce three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the cells are discharged from the first material output port 2006 through the first output flow channel 2005 to the outside of the chip and collected.

[0015] The acoustic fluidic unit 20 with a gap comprises a second right-angled triangle-shaped sharp corner 201 and a second right-angled edge 202, with a second cell capture flow channel 203 in the middle; the second cell capture flow channel 203 has a second capture flow channel inlet 2031 at the top, a third side wall 2032 below the left second right-angled triangle-shaped sharp corner 201, a fourth side wall 2033 and a second capture flow channel outlet 2034 below the right lower right-angled edge, a second capture flow channel bottom edge 2035 at the bottom, and a second cell capture flow channel. 203 is connected to the second main channel 204 above, the left side of the sharp corner 201 of the second right-angled triangle and the right side of the second right-angled edge 202 are the second connecting part 205 between units, the two right-angled sides of the sharp corner 201 of the second right-angled triangle are respectively located on the side close to the second main channel 204 and the side of the second connecting part 205, the thickness of the sharp corner 201 of the second right-angled triangle is smaller than the second right-angled edge 202 and the second connecting part 205 between units, and there is a movement gap 2011 at the bottom of the sharp corner 201 of the second right-angled triangle.

[0016] The second capture channel outlet 2034 of the second cell capture channel 203 in the gap acoustic fluidic unit 20 is arranged as one or more as needed, and is arranged respectively below the third side wall 2032 of the second cell capture channel 203, below the fourth side wall 2033 and at the bottom edge 2035 of the second capture channel.

[0017] The gap-type acoustic fluidic unit 20 is prepared into a microfluidic chip using a reverse molding process. When the material is poured into the unit pattern arranged in an array, the pattern will be covered, and an upper wall of a certain thickness will be formed above the unit. This thickness is used to lift the chip wafer with the unit pattern characteristics from the mold and divide it into independent chip devices.

[0018] The second open acoustic fluidic chip 300 based on the acoustic fluidic unit 20 with gaps includes the acoustic fluidic units 20 with gaps distributed in an array, the second capture flow channel outlet 2034 of each acoustic fluidic unit 20 with gaps is connected through the third pressure guide channel 3001, and is accumulated through the third pressure outlet 3002; negative pressure is applied at the third pressure outlet 3002, and negative pressure can be generated in the second cell capture flow channel 203 of each acoustic fluidic unit 20 with gaps through the third pressure guide channel 3001, thereby fixing the captured cells; the outer side of the acoustic fluidic unit 20 with gaps is the second main flow channel 204, and there is a second main flow channel 204 above the second main flow channel 204. The upper wall 2041 of the channel, and the outside of the second main channel is the second open fluid environment 3010; the target cells are placed in the second open fluid environment 3010 using a pipetting device, and when the sound waves are transmitted to the sharp corner 201 of the second right-angled triangle and the vicinity of the second cell capture channel 203, high-speed vortexes can be generated, causing micro-scale particles such as cells to rotate. When they rotate to a suitable angle, the capture function of the chip can be used to fix the target cells in the second cell capture channel 203, and the external operating tool moves in the second open fluid environment 3010 to the gap below the upper wall 2041 of the second main channel to contact the captured and fixed cells and carry out subsequent operations.

[0019] The second closed acoustic fluidic chip 400 based on the acoustic fluidic unit 20 with gaps includes the acoustic fluidic units 20 with gaps symmetrically distributed along the center line, the second capture flow channel outlet 2034 of each acoustic fluidic unit 20 with gaps is connected through the fourth pressure guide channel 4001, and is accumulated through the fourth pressure outlet 4002; applying negative pressure at the fourth pressure outlet 4002 can generate negative pressure in the second cell capture flow channel 203 of each acoustic fluidic unit 20 with gaps through the fourth pressure guide channel 4001, thereby fixing the captured cells; two acoustic fluidic units with gaps symmetrically distributed above and below The upper wall 2041 of the second main flow channel outside the fluidic unit 20 array forms a second closed operating environment 4010. The target cells are input into the second closed operating environment 4010 through the second material input port 4004 and the first input flow channel 2003. When the sound waves are transmitted to the sharp corner 201 of the second right-angled triangle and the vicinity of the second cell capture flow channel 203, high-speed eddy currents are generated, causing micro-scale particles such as cells to produce three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the cells are discharged from the second material output port 4006 through the second output flow channel 4005 to the outside of the chip and collected.

[0020] A chip system 500 having an asymmetric multi-angle acoustic fluidic unit, wherein the acoustic fluidic chip is selected from four types according to demand: a first open acoustic fluidic chip 100 based on a gapless acoustic fluidic unit 10, a first closed acoustic fluidic chip 200 based on a gapless acoustic fluidic unit 10, a second open acoustic fluidic chip 300 based on a gapped acoustic fluidic unit 20, and a second closed acoustic fluidic chip 400 based on a gapped acoustic fluidic unit 20; the chip is fixed on a substrate 50, and an acoustic vibration source 60 is provided outside the chip.

[0021] The beneficial effects of the present invention are:

[0022] 1) Compared with the jet generated by the bypass flow channel of the traditional microfluidic chip, the present invention adopts the principle of acoustic fluidic drive, which uses sound waves transmitted to a specific structure to generate high-frequency acoustic fluidic vortex. It has a simple structure, low preparation process difficulty, and faster cell rotation speed.

[0023] 2) Compared with the symmetrical triangle sharp corner structure adopted by the traditional acoustic fluidic chip, the present invention proposes a combination structure of an asymmetric right triangle sharp corner and a right-angled edge, which can produce a strong acoustic flow concentration phenomenon at the flow channel opening between the two. In addition, the present invention proposes a combination structure of an asymmetric right triangle sharp corner and a right-angled edge with a gap at the bottom, which can produce a stronger acoustic flow vortex when the sound wave is transmitted to the suspended asymmetric sharp corner. The gapless acoustic fluidic unit 10 and the gapped acoustic fluidic unit 20 proposed by the present invention will produce a higher cell rotation rate than the traditional acoustic fluidic cell rotation chip.

[0024] 3) The gapless acoustic fluidic unit 10 and the gapped acoustic fluidic unit 20 proposed in the present invention have cell capture and fixation functions that other chips do not have: the cell capture flow channel in the unit has interconnected pressure channels, and the cells captured and rotated by the sharp-angle unit can be fixed in situ by inputting negative pressure at the pressure input port, thereby facilitating subsequent cell manipulation.

[0025] 4) The open acoustofluidic chip proposed in this invention, featuring asymmetric multi-cornered cells, allows target cells to be released into an open environment and immobilized using the chip's capture function. Compared to traditional closed chips, external manipulation tools can access captured and immobilized cells within the open fluid environment, enabling a wider range of cell manipulation applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the gapless acoustic fluidic unit structure 10 of the present invention, (a) is a top view of the structure, and (b) is a schematic diagram of the A0-A0 cross-section of the sharp corner 101 of the right triangle shape and the main channel 104 in (a).

[0027] Figure 2This is a schematic diagram of an open acoustofluidic chip 100 based on a gapless acoustofluidic unit 10 of the present invention, wherein (a) is a top view of the structure, and (b) is a schematic diagram of the main channel 104, the upper wall 1041 of the main channel and the open environment 1010 in the A1-A1 section in (a).

[0028] Figure 3 This is a schematic diagram of a closed acoustofluidic chip 200 based on a gapless acoustofluidic unit 10 of the present invention, wherein (a) is a top view of the structure, and (b) is a schematic diagram of the main channel 104, the upper wall 1041 of the main channel and the closed environment 2010 in the A2-A2 section in (a).

[0029] Figure 4 Schematic diagram of the acoustic fluidic unit structure 20 with a gap according to the present invention, (a) is a top view of the structure, and (b) is a schematic diagram of the sharp corner 201 of the right triangle shape in (a), the moving gap 2011, and the main flow channel 204 at the B0-B0 section.

[0030] Figure 5 This is a schematic diagram of an open acoustic fluidic chip 300 based on a gap acoustic fluidic unit 20 according to the present invention, wherein (a) is a top view of the structure, and (b) is a schematic diagram of the main channel 204, the upper wall 2041 of the main channel and the open environment 3010 in the B1-B1 section in (a).

[0031] Figure 6 This is a schematic diagram of a closed acoustofluidic chip 400 based on a gap acoustofluidic unit 20 according to the present invention, wherein (a) is a top view of the structure, and (b) is a schematic diagram of the main channel 204, the upper wall 2041 of the main channel and the open environment 4010 in the B2-B2 section in (a).

[0032] Figure 7 Schematic diagram of the asymmetric multi-corner acoustofluidic chip system of the present invention. DETAILED DESCRIPTION

[0033] The present invention is described in detail below with reference to the accompanying drawings and embodiments.

[0034] An acoustofluidic chip based on an asymmetric multi-cusp structure, including an open acoustofluidic chip or a closed acoustofluidic chip based on a gapless acoustofluidic unit 10 or a gapped acoustofluidic unit 20;

[0035] Reference Figure 1The gapless acoustic fluidic unit 10 includes a first right-angled triangle-shaped sharp corner 101 and a first right-angled edge 102, with a first cell capture channel 103 located between the two. The first cell capture channel 103 has a first capture channel inlet 1031 at its top, a first sidewall 1032 below the left side of the first right-angled triangle-shaped sharp corner 101, a second sidewall 1033 and a first capture channel outlet 1034 below the right right-angled edge, and a first capture channel bottom edge 1035 at its bottom. The first cell capture channel 103 is connected to the first main channel 104 above. The left side of the first right-angled triangle sharp corner 101 and the right side of the first right-angled edge 102 form a first inter-unit connection portion 105. The two right-angled sides of the first right-angled triangle sharp corner 101 are located on the side close to the first main channel 104 and the side close to the first connection portion 105, respectively. The first right-angled triangle sharp corner 101 has the same thickness as the first right-angled edge 102 and the first connection portion 105.

[0036] The first capture channel outlet 1034 in the gapless acoustic fluidic unit 10 is arranged as one or more as needed, and is arranged below the first side wall 1032 of the first cell capture channel 103, below the second side wall 1033 and at the bottom edge 1035 of the first capture channel.

[0037] The gapless acoustic fluidic unit 10 is prepared into a microfluidic chip using a process including but not limited to a reverse molding process. The material poured into the unit pattern arranged in an array will cover the pattern and form an upper wall of a certain thickness above the unit. This thickness can be used to lift the chip wafer with the unit pattern characteristics from the mold and divide it into independent chip devices.

[0038] Reference Figure 2The first open acoustic fluidic chip 100 based on the gapless acoustic fluidic unit 10 includes the gapless acoustic fluidic units 10 distributed in an array, wherein the first capture flow channel outlets 1034 of each gapless acoustic fluidic unit 10 are connected through a first pressure guide channel 1001 and accumulated through a first pressure outlet 1002; negative pressure is applied at the first pressure outlet 1002, which can generate negative pressure in the first cell capture flow channel 103 of each gapless acoustic fluidic unit 10 through the first pressure guide channel 1001, thereby fixing the captured cells; the outer side of the gapless acoustic fluidic unit 10 is a first main flow channel 104, and above the first main flow channel 104 there is a first main flow channel upper wall 10 41. The outside of the first main channel 104 is a first open fluid environment 1010. Target cells can be placed in the first open fluid environment 1010 using a pipette or other pipetting equipment. When the sound waves are transmitted to the sharp corner 101 of the first right-angled triangle and the first cell capture channel 103, high-speed vortices can be generated, causing micro-scale particles such as cells to produce three-dimensional rotational motion. When the target cells are rotated to a suitable angle, the capture function of the chip can be used to fix them in the first cell capture channel 103. External operating tools can be moved from the first open fluid environment 1010 to the gap below the upper wall 1041 of the first main channel to contact the captured and fixed cells and carry out subsequent operations.

[0039] Reference Figure 3 The first closed acoustofluidic chip 200 based on the gapless acoustofluidic unit 10 includes an array of gapless acoustofluidic units 10 symmetrically distributed along the midline, wherein the first cell capture flow channel 103 of each gapless acoustofluidic unit 10 is connected through a second pressure guide channel 2001 and accumulated through a second pressure outlet 2002; negative pressure is applied at the second pressure outlet 2002, which can generate negative pressure in the first cell capture flow channel 103 of each gapless acoustofluidic unit 10 through the second pressure guide channel 2001, thereby fixing the captured cells; two gapless acoustofluidic units 10 symmetrically distributed above and below The upper wall 1041 of the first main flow channel outside the array of acoustic fluidic control units 10 forms a first closed operating environment 2010. The target cells are input into the first closed operating environment 2010 through the first material input port 2004 via the first input flow channel 2003. When the sound waves are transmitted to the sharp corner 101 of the first right-angled triangle and the first cell capture flow channel 103, high-speed vortices are generated, causing micro-scale particles such as cells to produce three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the cells are discharged from the first material output port 2006 through the first output flow channel 2005 to the outside of the chip and collected.

[0040] Reference Figure 4The acoustic fluidic unit 20 with a gap comprises a second right-angled triangle-shaped sharp corner 201 and a second right-angled edge 202, with a second cell capture flow channel 203 in the middle; the second cell capture flow channel 203 has a second capture flow channel inlet 2031 at the top, a third side wall 2032 below the left second right-angled triangle-shaped sharp corner 201, a fourth side wall 2033 and a second capture flow channel outlet 2034 below the right lower right-angled edge, a second capture flow channel bottom edge 2035 at the bottom, and a second cell capture flow channel inlet 2031 at the top. Channel 203 is connected to the second main channel 204 above. The left side of the sharp corner 201 of the second right-angled triangle and the right side of the second right-angled edge 202 are the second connecting part 205 between units. The two right-angled sides of the sharp corner 201 of the second right-angled triangle are respectively located on the side close to the second main channel 204 and the side of the second connecting part 205. The thickness of the sharp corner 201 of the second right-angled triangle is smaller than the second right-angled edge 202 and the second connecting part 205 between units. There is a movement gap 2011 at the bottom of the sharp corner 201 of the second right-angled triangle.

[0041] The second capture channel outlet 2034 of the second cell capture channel 203 in the gap acoustic fluidic unit 20 can be arranged as one or more as needed, and respectively arranged below the third side wall 2032 of the second cell capture channel 203, below the fourth side wall 2033 and at the bottom edge 2035 of the second capture channel.

[0042] The gap-type acoustic fluidic unit 20 can be prepared into a microfluidic chip using a process including but not limited to a reverse molding process. The material poured into the unit pattern arranged in an array will cover the pattern and form an upper wall of a certain thickness above the unit. This thickness can be used to lift the chip wafer with the unit pattern characteristics from the mold and divide it into independent chip devices.

[0043] Reference Figure 5, a second open acoustic fluidic chip 300 based on the acoustic fluidic unit 20 with gaps: including the acoustic fluidic units 20 with gaps distributed in an array, the second capture flow channel outlet 2034 of each acoustic fluidic unit 20 with gaps is connected through a third pressure guide channel 3001, and accumulated through the third pressure outlet 3002; applying negative pressure at the third pressure outlet 3002 can generate negative pressure in the second cell capture flow channel 203 of each acoustic fluidic unit 20 with gaps through the third pressure guide channel 3001, thereby fixing the captured cells; the outer side of the acoustic fluidic unit 20 with gaps is the second main flow channel 204, and there is a second main flow channel upper wall above the second main flow channel 204. 2041, the outside of the second main channel is a second open fluid environment 3010; target cells can be placed in the second open fluid environment 3010 using a pipette or other pipetting equipment, and when the sound waves are transmitted to the sharp corner 201 of the second right-angled triangle and the vicinity of the second cell capture channel 203, high-speed vortexes can be generated, causing micro-scale particles such as cells to rotate. When they rotate to a suitable angle, the capture function of the chip can be used to fix the target cells in the second cell capture channel 203, and external operating tools can be moved in the second open fluid environment 3010 to the gap below the upper wall 2041 of the second main channel to contact the captured and fixed cells and carry out subsequent operations.

[0044] Reference Figure 6 , a second closed acoustic fluidic chip 400 based on the acoustic fluidic unit 20 with gaps: comprising acoustic fluidic units 20 with gaps symmetrically distributed along the midline, wherein the second capture flow channel outlets 2034 of each acoustic fluidic unit 20 with gaps are connected through a fourth pressure guide channel 4001 and accumulated through a fourth pressure outlet 4002; applying negative pressure at the fourth pressure outlet 4002 can generate negative pressure in the second cell capture flow channel 203 of each acoustic fluidic unit 20 with gaps through the fourth pressure guide channel 4001, thereby fixing the captured cells; two acoustic fluidic units with gaps symmetrically distributed above and below The upper wall 2041 of the second main flow channel outside the fluidic unit 20 array forms a second closed operating environment 4010. The target cells are input into the second closed operating environment 4010 through the second material input port 4004 and the first input flow channel 2003. When the sound waves are transmitted to the sharp corner 201 of the second right-angled triangle and the vicinity of the second cell capture flow channel 203, high-speed eddy currents are generated, causing micro-scale particles such as cells to produce three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the cells are discharged from the second material output port 4006 through the second output flow channel 4005 to the outside of the chip and collected.

[0045] The number of gapless acoustic fluidic units 10 and gapped acoustic fluidic units 20 used in the first open acoustic fluidic chip 100 based on the gapless acoustic fluidic unit 10, the first closed acoustic fluidic chip 200 based on the gapless acoustic fluidic unit 10, the second open acoustic fluidic chip 300 based on the gapped acoustic fluidic unit 20, and the second closed acoustic fluidic chip 400 based on the gapped acoustic fluidic unit 20 can be changed according to needs, and the shape of the open fluid environment or closed fluid environment outside the unit can use, but is not limited to, the rectangle used in the figure, and its shape and size can be changed according to application requirements; the main flow channel, pressure channel, raw material input channel and input port, raw material output channel and output port in the chip can use, but is not limited to, the rectangle used in the figure, and its shape and size can be changed according to application requirements.

[0046] Reference Figure 7 A chip system 500 with an asymmetric multi-angle acoustic fluidic unit, wherein the acoustic fluidic chip can be selected from four types according to needs: a first open acoustic fluidic chip 100 based on a gapless acoustic fluidic unit 10, a first closed acoustic fluidic chip 200 based on a gapless acoustic fluidic unit 10, a second open acoustic fluidic chip 300 based on a gapped acoustic fluidic unit 20, and a second closed acoustic fluidic chip 400 based on a gapped acoustic fluidic unit 20; the chip is fixed on a substrate 50, and an acoustic vibration source 60 is provided outside the chip.

[0047] The first open acoustic fluidic chip 100 based on the gapless acoustic fluidic unit 10, the first closed acoustic fluidic chip 200 based on the gapless acoustic fluidic unit 10, the second open acoustic fluidic chip 300 based on the gapped acoustic fluidic unit 20, and the second closed acoustic fluidic chip 400 based on the gapped acoustic fluidic unit 20 can be prepared using materials including but not limited to PDMS, and the chips can be stably fixed on the substrate 50 using a bonding process.

[0048] The substrate 50 may be made of, but not limited to, a glass slide, a culture dish, or any other flat surface base material made of glass, plastic, or other piezoelectric materials.

[0049] The acoustic vibration source 60 includes but is not limited to any excitation device that can generate surface acoustic waves, such as an interdigital transducer electrode (IDT) or a piezoelectric ceramic resonator. Applying alternating signals of different frequencies and amplitudes on such a device can generate specific acoustic wave vibration signals in the substrate. When the sound source of specific frequency and amplitude is transmitted to each gapless acoustic fluidic unit 10 and gapped acoustic fluidic unit 20 inside the chip, an enhanced rotating acoustic flow can be generated at the first capture flow channel inlet 1031 and the second capture flow channel inlet 2031, thereby realizing the three-dimensional rotation of micro-scale particles such as cells.

[0050] One or more acoustic vibration sources 60 can be arranged around the microfluidic chip as needed.

Claims

1. An acoustofluidic chip based on an asymmetric multi-cusp structure, comprising an open acoustofluidic chip or a closed acoustofluidic chip based on a gapless acoustofluidic unit (10) or a gapped acoustofluidic unit (20); The gapless acoustic fluidic unit (10) comprises a first right-angled triangle-shaped sharp corner (101) and a first right-angled edge (102), with a first cell capture flow channel (103) located between the two. The first cell capture flow channel (103) has a first capture flow channel inlet (1031) at the top, a first side wall (1032) below the left first right-angled triangle-shaped sharp corner (101), a second side wall (1033) and a first capture flow channel outlet (1034) below the right lower right-angled edge, and a first capture flow channel bottom edge (1031) at the bottom. 35); the first cell capture flow channel (103) is connected to the first main flow channel (104) above, the left side of the sharp corner (101) of the first right-angled triangle and the right side of the first right-angled edge (102) are the first connecting portion (105) between the units, and the two right-angled sides of the sharp corner (101) of the first right-angled triangle are respectively located on the side close to the first main flow channel (104) and the side of the first connecting portion (105); the sharp corner (101) of the first right-angled triangle has the same thickness as the first right-angled edge (102) and the first connecting portion (105); One or more first capture flow channel outlets (1034) in the gapless acoustic fluidic unit (10) are arranged as needed, and are respectively arranged below the first side wall (1032) and below the second side wall (1033) of the first cell capture flow channel (103) and at the bottom edge (1035) of the first capture flow channel; The acoustic fluidic unit (20) with a gap comprises a second right-angled triangle-shaped sharp corner (201) and a second right-angled edge (202), wherein a second cell capture flow channel (203) is located between the two; the second cell capture flow channel (203) has a second capture flow channel inlet (2031) at the upper portion, a third side wall (2032) below the left second right-angled triangle-shaped sharp corner (201), a fourth side wall (2033) and a second capture flow channel outlet (2034) below the right lower right-angled edge, a second capture flow channel bottom edge (2035) at the bottom, and a second cell capture flow channel (2031) at the lower portion. 03) is connected to the second main flow channel (204) above, the left side of the sharp corner (201) of the second right-angled triangle and the right side of the second right-angled edge (202) are the second connecting part (205) between units, the two right-angled sides of the sharp corner (201) of the second right-angled triangle are respectively located on the side close to the second main flow channel (204) and the side of the second connecting part (205), the thickness of the sharp corner (201) of the second right-angled triangle is smaller than the second right-angled edge (202) and the second connecting part (205) between units, and there is a movement gap (2011) at the bottom of the sharp corner (201) of the second right-angled triangle.

2. The acoustofluidic chip according to claim 1, characterized in that: The gapless acoustic fluidic unit (10) is prepared into a microfluidic chip using a reverse molding process. When the material is poured into the unit pattern arranged in an array, the pattern is covered and an upper wall with a certain thickness is formed above the unit. The chip wafer with the unit pattern characteristics is lifted from the mold using the thickness and divided into independent chip devices.

3. The acoustofluidic chip according to claim 1, characterized in that: A first open acoustic fluidic chip (100) based on a gapless acoustic fluidic unit (10) comprises an array of gapless acoustic fluidic units (10), wherein the first capture flow channel outlets (1034) of each gapless acoustic fluidic unit (10) are connected via a first pressure guide channel (1001) and accumulated via a first pressure outlet (1002); negative pressure is applied at the first pressure outlet (1002), thereby generating negative pressure in the first cell capture flow channel (103) of each gapless acoustic fluidic unit (10) via the first pressure guide channel (1001), thereby fixing the captured cells; the outer side of the gapless acoustic fluidic unit (10) is a first main flow channel (104), and a first main flow channel upper wall (1041) is provided above the first main flow channel (104), and the first main flow channel upper wall (1041) is provided above the first main flow channel (104). The outside of a main channel (104) is a first open fluid environment (1010), and target cells are placed in the first open fluid environment (1010) using a pipetting device. When the sound wave is transmitted to the sharp corner (101) of the first right-angled triangle and the vicinity of the first cell capture channel (103), a high-speed vortex is generated, causing micro-scale particles containing cells to produce three-dimensional rotational motion. When the target cells are rotated to the corresponding angle, the capture function of the chip is used to fix them at the first cell capture channel (103). An external operating tool moves in the first open fluid environment (1010) to the gap below the upper wall (1041) of the first main channel to contact the captured and fixed cells and carry out subsequent operations. The number of arrays of gapless acoustic fluidic control units (10) in the chip is set according to application requirements.

4. The acoustofluidic chip according to claim 1, wherein: A first closed acoustofluidic chip (200) based on a gapless acoustofluidic unit (10) comprises an array of gapless acoustofluidic units (10) symmetrically distributed along a midline, wherein the first cell capture flow channel (103) of each gapless acoustofluidic unit (10) is connected via a second pressure guide channel (2001) and accumulated via a second pressure outlet (2002); negative pressure is applied at the second pressure outlet (2002), thereby generating negative pressure in the first cell capture flow channel (103) of each gapless acoustofluidic unit (10) via the second pressure guide channel (2001), thereby fixing the captured cells; two gapless acoustofluidic units (10) symmetrically distributed above and below The upper wall (1041) of the first main flow channel outside the array of the gap acoustic fluidic control unit (10) forms a first closed operating environment (2010). The target cells are input into the first closed operating environment (2010) through the first material input port (2004) via the first input flow channel (2003). When the sound waves are transmitted to the sharp corner (101) of the first right-angled triangle and the vicinity of the first cell capture flow channel (103), high-speed eddy currents are generated, causing micro-scale particles containing cells to generate three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the particles are discharged from the first material output port (2006) through the first output flow channel (2005) to the outside of the chip and collected. The number of arrays of gapless acoustofluidic units (10) within the chip is set according to application requirements.

5. The acoustofluidic chip according to claim 1, characterized in that: The second capture flow channel outlet (2034) of the second cell capture flow channel (203) in the gap acoustic fluidic unit (20) is arranged one or more as needed, and is respectively arranged below the third side wall (2032) of the second cell capture flow channel (203), below the fourth side wall (2033) and at the bottom edge (2035) of the second capture flow channel.

6. The acoustofluidic chip according to claim 1, characterized in that: The gapped acoustic fluidic unit (20) is prepared into a microfluidic chip using a reverse molding process. When the material is poured into the unit pattern arranged in an array, the pattern is covered and an upper wall with a certain thickness is formed above the unit. The chip wafer with the unit pattern characteristics is lifted from the mold using the thickness and divided into independent chip devices.

7. The acoustofluidic chip according to claim 1, characterized in that: A second open acoustic fluidic chip (300) based on an acoustic fluidic unit with gaps (20) comprises acoustic fluidic units (20) with gaps distributed in an array, wherein the second capture flow channel outlets (2034) of each acoustic fluidic unit with gaps (20) are connected via a third pressure guide channel (3001) and accumulated via a third pressure outlet (3002); negative pressure is applied at the third pressure outlet (3002), and negative pressure can be generated in the second cell capture flow channel (203) of each acoustic fluidic unit with gaps (20) via the third pressure guide channel (3001), thereby fixing the captured cells; the outer side of the acoustic fluidic unit with gaps (20) is a second main flow channel (204), and above the second main flow channel (204) there is a second main flow channel upper wall (204) 1), the outside of the second main channel is a second open fluid environment (3010); target cells are placed in the second open fluid environment (3010) using a pipetting device, and when the sound wave is transmitted to the sharp corner (201) of the second right-angled triangle and the vicinity of the second cell capture channel (203), a high-speed vortex is generated, causing the micro-scale particles containing the cells to produce a rotational motion, and when the target cells are rotated to the corresponding angle, the capture function of the chip is used to fix the target cells at the second cell capture channel (203), and an external operating tool moves in the second open fluid environment (3010) to the gap below the upper wall (2041) of the second main channel to contact the captured and fixed cells and carry out subsequent operations; the number of arrays of gap acoustic fluidic control units (20) in the chip is set according to application requirements.

8. The acoustofluidic chip according to claim 1, characterized in that: The second closed acoustofluidic chip (400) based on the acoustofluidic unit (20) with gaps comprises the acoustofluidic units (20) with gaps symmetrically distributed along the midline, wherein the second capture flow channel outlets (2034) of each acoustofluidic unit (20) with gaps are connected via a fourth pressure guide channel (4001) and accumulated via a fourth pressure outlet (4002); negative pressure is applied at the fourth pressure outlet (4002), thereby generating negative pressure in the second cell capture flow channel (203) of each acoustofluidic unit (20) with gaps through the fourth pressure guide channel (4001), thereby fixing the captured cells; two acoustofluidic units (20) with gaps symmetrically distributed above and below The upper wall (2041) of the second main flow channel outside the array of the acoustofluidic unit (20) forms a second closed operating environment (4010). The target cells are input into the second closed operating environment (4010) through the second material input port (4004) via the first input flow channel (2003). When the sound waves are transmitted to the sharp corner (201) of the second right-angled triangle and the vicinity of the second cell capture flow channel (203), high-speed eddy currents are generated, causing micro-scale particles containing cells to generate three-dimensional rotational motion and perform subsequent operations. After the operation is completed, the particles are discharged from the second material output port (4006) through the second output flow channel (4005) to the outside of the chip and collected. The number of arrays of gap acoustofluidic units (20) in the chip is set according to application requirements.

9. A chip system (500) having an asymmetric multi-cusp acoustofluidic unit, characterized in that: The acoustic fluidic chip based on any one of claims 1 to 8 is selected according to demand from four types: a first open acoustic fluidic chip (100) based on a gapless acoustic fluidic unit (10), a first closed acoustic fluidic chip (200) based on a gapless acoustic fluidic unit (10), a second open acoustic fluidic chip (300) based on a gapped acoustic fluidic unit (20), and a second closed acoustic fluidic chip (400) based on a gapped acoustic fluidic unit (20); the chip is fixed on a substrate (50), and an acoustic vibration source (60) is provided outside the chip.

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