Separation device and separation method

Abstract

The separation device (100) includes a separation chip (10, 10'), a vibration system (70), a differential pressure driving system (50), a frequency conversion module (40), and a controller (60). The separation chip (10, 10') includes a sample pool (11), a first chamber (12, 12') and a second chamber (13, 13') located on both sides of the sample pool (11). The vibration system (70) includes a compression assembly (20) and two ultrasonic generators (30), and the compression assembly (20) drives the two ultrasonic generators (30) to move toward the separation chip so that they are coupled to the outer surface of the first chamber (12, 12') and the outer surface of the second chamber (13, 13'), respectively. The frequency conversion module (40) controls the differential pressure driving system (50) to alternately generate differential pressure in the first chamber (12, 12') and the second chamber (13, 13'). The controller (60) controls the vibration of the two ultrasonic generators (30) when the differential pressure driving system (50) stops generating differential pressure in the first chamber (12, 12') or the second chamber (13, 13'). The separation device (100) allows for convenient assembly of the ultrasonic generator (30) and the separation chip (10, 10'), has low cost for the separation chip (10, 10'), high separation efficiency, and can effectively prevent clogging of the filter membrane. Additionally, a separation method is further provided.

Description

Technology Field

[0001] The present invention relates to the field of exosome separation technology, and specifically to a separation device and a separation method for separating target particles from a liquid sample. Background Technology

[0002] Exosomes are small vesicles with a double phospholipid membrane structure and a diameter of 30–150 nm that are continuously secreted in large quantities by living cells. As carriers for intercellular communication, they transport specific components such as proteins, nucleic acids, and metabolic small molecules derived from parent cells. Numerous studies indicate that exosomes are involved in various events of tumor development, including immune evasion, angiogenesis, tumor metastasis, and tumor resistance. Exosomes can be continuously released from cancer cells early on and enter the patient's bloodstream, and their lipid bilayer structure effectively protects the transported proteins and encapsulated nucleic acid materials. Exosomes are widely and stably present in various clinical samples, including blood, urine, ascites fluid, tissue fluid, tears, saliva, and cerebrospinal fluid. Among these, blood and urine are easy to clinically sample due to their high exosome counts. Therefore, exosomes are considered a key research subject in the fields of in vitro diagnostic research and oncology clinical trials, and are expected to demonstrate great clinical value in terms of early diagnosis of tumors, evaluation of tumor metastasis and recurrence, evaluation of tumor heterogeneity, dynamic detection of tumor development and therapeutic effects, detection of drug resistance mutations, and personalized medicine.

[0003] Currently, the main obstacle to realizing the clinical application of exosomes is how to separate exosomes and their subgroups from complex biological fluids. Existing separation processes have disadvantages such as being cumbersome, taking a long time to separate, having low separation efficiency, easily clogging filtration membranes, low separation flux, and low purity.

[0004] It is necessary to propose a separation device to resolve the disadvantages of the prior art as described above.

[0005] In addition, the present invention further provides a separation method for separating exosomes using the separation device.

[0006] According to the first embodiment, a separation device for separating and purifying target particles from a liquid sample is provided, said separation device comprises a separation chip, a vibration system, a differential pressure driving system, a frequency conversion module, and a controller. The separation chip comprises a sample pool, at least one first chamber located on one side of the sample pool, and at least one second chamber located on the other side of the sample pool away from the at least one first chamber, and the two adjacent first chambers and the first chamber adjacent to the sample pool and the sample pool are all connected through a first filter membrane, and the two adjacent second chambers and the second chamber adjacent to the sample pool and the sample pool are all connected through a second filter membrane, and the diameter of each of the first filter membranes and each of the second filter membranes is smaller than the particle diameter of the target particles. The vibration system comprises a compression assembly and two ultrasonic generators located in the compression assembly. The compression assembly drives the ultrasonic generator to move toward the separation chip, thereby causing the two ultrasonic generators to be coupled to the outer surface of the outermost first chamber and the outer surface of the outermost second chamber, respectively, in close proximity to the cross-section of the separation chip. The frequency conversion module is connected to the first chamber and the second chamber, respectively, through the differential pressure driving system. The frequency conversion module controls the differential pressure driving system to alternately generate differential pressure in the first chamber and the second chamber. When the differential pressure driving system stops generating differential pressure in the first chamber or the second chamber, the controller controls the vibration of the two ultrasonic generators to generate ultrasonic vibration waves, respectively.

[0007] In an embodiment of the present invention, the vibration frequencies of the two ultrasonic vibration waves are both 15 KHz to 80 KHz; and within one cycle of the ultrasonic vibration waves, the duty ratio of the two ultrasonic vibration waves is both 10% to 90%.

[0008] In an embodiment of the present invention, the difference between the vibration frequency and the duty ratio of the two ultrasonic vibration waves is 30% or less.

[0009] In an embodiment of the present invention, within one cycle of turning on or off of the two ultrasonic generators, the ratio of turning on is 10% to 100%.

[0010] In an embodiment of the present invention, the two ultrasonic generators are located on the same horizontal plane.

[0011] In an embodiment of the present invention, the ultrasonic generator comprises an amplitude transformer installed in close proximity to the separation chip, and a piezoelectric ceramic assembly connected to the amplitude transformer.

[0012] In an embodiment of the present invention, the amplitude transformer comprises a first amplitude variation part, a second amplitude variation part, a third amplitude variation part, and a connecting part that are sequentially connected. The connecting part is connected to the piezoelectric ceramic assembly, and the first amplitude variation part is coupled to the outer surface of the first chamber or the outer surface of the second chamber, separated from the cross-section of the second amplitude variation part. Along a direction perpendicular to the extension direction of the amplitude transformer, the size of the second amplitude variation part is made smaller than the size of the first amplitude variation part and the third amplitude variation part.

[0013] In an embodiment of the present invention, the piezoelectric ceramic assembly comprises a plurality of piezoelectric ceramic sheets installed in a stacked manner, a plurality of electrode pieces installed spaced apart from the plurality of piezoelectric ceramic sheets, an insulating sleeve, and a connecting member, wherein the plurality of piezoelectric ceramic sheets and the plurality of electrode pieces are covered and installed on the insulating sleeve, and the connecting member penetrates the insulating sleeve and is detachably connected to the amplitude transformer.

[0014] In an embodiment of the present invention, the ultrasonic generator further comprises a control block, the control block is installed at one end of the piezoelectric ceramic assembly away from the amplitude transformer, and the connecting member is connected to the control block.

[0015] According to the second aspect, a separation method for separating and purifying target particles from a liquid sample is provided, said method,

[0016] A step of providing a separation device as described above and providing a liquid sample to the sample pool;

[0017] A step of controlling the compression assembly to drive the two ultrasonic generators to move toward the separation chip, so that the two ultrasonic generators are coupled to the outer surface of the outermost first chamber and the outer surface of the outermost second chamber, respectively, in close proximity to the cross-section of the separation chip;

[0018] A step of generating a differential pressure in at least one first chamber so that a component of the liquid sample in the sample pool, having a particle size smaller than the diameter of the first filter membrane, enters the at least one first chamber under the action of the differential pressure;

[0019] A step of stopping the generation of differential pressure in at least one first chamber and controlling the vibration of two ultrasonic generators to generate ultrasonic vibration waves;

[0020] A step of generating a differential pressure in at least one second chamber so that a component of the liquid sample in the sample pool, having a particle size smaller than the diameter of the second filter membrane, enters the at least one second chamber under the action of the differential pressure; and

[0021] The method includes the step of stopping the generation of differential pressure in at least one second chamber and controlling the vibration of two ultrasonic generators to generate ultrasonic vibration waves.

[0022] The separation device of the present invention separately installs a separation chip and an ultrasonic generator, moves two ultrasonic generators to be coupled to the separation chip through the interlocking of a compression assembly during use, and provides ultrasonic vibration waves to the separation chip to vibrate the separation chip, thereby preventing clogging of the first filter membrane and the second filter membrane, and improving separation efficiency and separation purity; and since there is no need to install a vibration member on the separation chip, the difficulty and cost of manufacturing the separation chip can be reduced.

[0023] The ultrasonic generator has a specific structure and parameters that match the separation chip, and the generated ultrasonic vibration waves can ensure that particles attached to the first and second filtration membranes are removed, thereby preventing clogging of the filtration membranes and improving filtration efficiency and filtration purity.

[0024] The compression assembly can implement the interlocking of two ultrasonic generators. This allows the two ultrasonic generators to be coupled to or separated from the separation chip, and the interlocking process is simple to operate. Since the ultrasonic generators and the separation chip are tightly coupled, it is possible to ensure that ultrasonic vibration waves are transmitted to the first filter membrane and the second filter membrane without damaging the separation chip. Brief explanation of the drawing

[0025] To more clearly explain the technical solution means of the embodiments of the present invention, the drawings to be used in the embodiments of the present invention are briefly described below; however, the drawings in the description below are merely some embodiments of the present invention, and it is obvious to those skilled in the art that other drawings can be obtained based on these drawings without the need to strive to create inventive step. Figure 1 is a schematic diagram of the structure of a separation chip provided by the prior art. FIG. 2 is a schematic diagram of a separation device provided by one embodiment of the present invention. FIG. 3 is a schematic diagram of a structure in which a separation chip, a vibration system, and a differential pressure driving system are connected, provided by one embodiment of the present invention. FIG. 4 is a schematic diagram of a structure combining a separation chip and an ultrasonic generator provided by one embodiment of the present invention. FIG. 5 is a schematic diagram of the structure of an ultrasonic generator provided by one embodiment of the present invention. FIG. 6 is a cross-sectional view of an ultrasonic generator provided by one embodiment of the present invention. FIGS. 7a to 7d are drawings showing photographs (images) of the vibration wave generation process of an ultrasonic generator provided by an embodiment of the present invention. FIG. 8 is a schematic diagram of the structure of a vibration system and a differential pressure driving system provided by one embodiment of the present invention. FIG. 9 is a schematic diagram of the structure of a separation chip provided by another embodiment of the present invention. The present invention is further explained by combining the specific embodiments below with the drawings above. Specific details for implementing the invention

[0026] Hereinafter, the technical solution means of the embodiments of the present invention will be clearly and completely explained with reference to the drawings of the embodiments of the present invention. Of course, the described embodiments are only some embodiments of the present invention and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by a person skilled in the art without needing to strive to create inventive step are all within the scope of protection of the present invention.

[0027] It should be noted that when a component is described as being "fixed" or "mounted" to another component, it may be located directly on the other component or an intermediate component may exist. When one component is considered to be "installed" on another component, it may be installed directly on the other component or an intermediate component may exist simultaneously. As used herein, the term "and / or" includes all combinations and any combination of one or more related listed items.

[0028] As illustrated in FIG. 1, the separation chip (10') comprises a sample pool (11'), and a first chamber (12') and a second chamber (13') located on opposite sides of the sample pool (11'). The first chamber (12') and the sample pool (11') are connected through a first filter membrane (14'), and the second chamber (13') and the sample pool (11') are connected through a second filter membrane (15'). The sample pool (11') accommodates a liquid sample. The diameters of the first filter membrane (14') and the second filter membrane (15') are smaller than the diameter of the target particle. A first opening (121') is installed in the first chamber (12'), and the first opening (121') is connected to the outside of the first chamber (12'). A second opening (131') is installed in the second chamber (13'), and the second opening (131') communicates with the outside of the second chamber (13'). In order to solve the problem of low filtration efficiency and easy clogging of the filter membrane as described above, a vibrating member (16') is fixed to the outer surface of the first chamber (12') and the second chamber (13') of the separation chip (10'), and to the first filter membrane (14') and the second filter membrane (15'), and clogging of the filter membrane is prevented through the vibration of the vibrating member (16'), thereby achieving the purpose of improving filtration efficiency, flux, purity, etc. However, mounting a vibration member (16') on the separation chip (10'), especially mounting a vibration member (16') on the first filter membrane (14') and the second filter membrane (15'), is very difficult, so circuit wiring control is difficult, there is a risk of short circuit, and the vibration member (16') is prone to falling off during the vibration process, affecting usage and making mass production difficult, and at the same time, the cost of the disposable separation chip (10') also increases.

[0029] Referring to FIGS. 2 through 4, in order to solve the above-mentioned problem of a separation chip (10'), one embodiment of the present invention provides a separation device (100), said separation device (100) includes a separation chip (10), a vibration system (70), a differential pressure driving system (50), a frequency conversion module (40), and a controller (60). The separation chip (10) includes a sample pool (11), at least one first chamber (12) located on one side of the sample pool (11), and at least one second chamber (13) located on the other side of the sample pool (11) away from the at least one first chamber (12). The two adjacent first chambers (12) and the first chamber (12) adjacent to the sample pool (11) and the sample pool (11) are all connected through a first filter membrane (14), and the two adjacent second chambers (13) and the second chamber (13) adjacent to the sample pool (11) and the sample pool (11) are all connected through a second filter membrane (15). The diameter of each of the first filter membrane (14) and each of the second filter membranes (15) is smaller than the diameter of the target particle. The vibration system (70) includes a compression assembly (20) and two ultrasonic generators (30) located in the compression assembly (20). The compression assembly (20) drives the two ultrasonic generators (30) to move toward the separation chip (10), thereby causing the two ultrasonic generators (30) to be coupled to the outer surface of the outermost first chamber (12) and the outer surface of the outermost second chamber (13), respectively, in close proximity to the cross-section of the separation chip (10). The frequency conversion module (40) is connected to the first chamber (12) and the second chamber (13), respectively, through the differential pressure driving system (50). The frequency conversion module (40) controls the differential pressure driving system (50) to alternately generate differential pressure in the first chamber (12) and the second chamber (13).The controller (60) controls the vibration of the two ultrasonic generators (30) to generate ultrasonic vibration waves when the differential pressure driving system (50) stops generating differential pressure in the first chamber (12) or the second chamber (13). The present invention allows the separation chip (10) and the ultrasonic generator (30) to be installed separately, and when in use, the two ultrasonic generators (30) are moved to be coupled to the separation chip (10). By providing ultrasonic vibration waves to the separation chip (10) to vibrate the separation chip (10), the first filter membrane (14) and the second filter membrane (15) can be prevented from clogging, and the separation efficiency and separation purity can be improved. In addition, since there is no need to install a vibration member in the separation chip (10), the difficulty and cost of manufacturing the separation chip (10) can be reduced.

[0030] Referring to FIGS. 4 to 6, the ultrasonic generator (30) includes an amplitude transformer (31) installed in close proximity to the separation chip (10), and a piezoelectric ceramic assembly (32) connected to the amplitude transformer (31). The controller (60) is connected to the piezoelectric ceramic assembly (32) to enable signal transmission and controls the opening and closing of the piezoelectric ceramic assembly (32) to generate ultrasonic vibration waves in the transverse direction (i.e., the longitudinal direction of the amplitude transformer (31). The ultrasonic vibration waves in the transverse direction are transmitted to the entire separation chip (10) through the outer walls of the first chamber (12) and the second chamber (13), causing the separation chip (10) to vibrate at a high frequency, thereby causing the first filter membrane (14) and the second filter membrane (15) to vibrate at a high frequency. Two ultrasonic vibration waves together disturb the liquid sample to generate acoustic streaming, thereby preventing the pores of the filter membrane from being clogged by target particles or the target particles from aggregating. Additionally, by allowing target particles adsorbed to the pores of the filter membrane to be rapidly separated from the pores and resuspended in the refluxed liquid sample, the clogging of the pores of the filter membrane can be prevented and high-efficiency separation can be achieved.

[0031] In one embodiment, to achieve the above purpose, the vibration frequency of the ultrasonic vibration waves generated by two ultrasonic generators (30) is 15 KHz to 80 KHz, and within one cycle of the ultrasonic vibration waves, the duty ratio of the two ultrasonic vibration waves is 10% to 90%. Controlling the vibration frequency and duty ratio of the two ultrasonic vibration waves within the above range can directly affect the magnitude of the total power actually output, and accordingly, affect the overall vibration effect and the degree of damage to the target particles. The total power of the ultrasonic vibration waves actually output within this frequency range and duty ratio range is suitable for penetrating the shell of the separation chip (10) and being transmitted to the first filter membrane (14) and the second filter membrane (15) inside. Thus, by vibrating the filter membrane with a constant amplitude, the target particles adsorbed on the filter membrane can be separated more quickly and returned to the liquid sample, and at the same time, by disturbing the liquid sample by the ultrasonic vibration waves, acoustic flow can be generated to disperse the aggregated particles. As a result, clogging of the filter membrane can be prevented and filtration efficiency can be improved. In addition, since the total power of the ultrasonic vibration waves output within the above frequency and duty cycle range is appropriate, the separation and purification quality can be guaranteed without causing damage to the target particles.

[0032] In one embodiment, the difference between the vibration frequency and the duty cycle of the two ultrasonic vibration waves is 30% or less, and since two ultrasonic generators (30) are installed on opposite sides of the separation chip (10), the direction of transmission of the generated ultrasonic vibration waves is reversed. By making the vibration frequency and duty cycle of the two ultrasonic vibration waves similar, the separation chip (10) can be made more stable during the vibration process, and by making the acoustic flow generated by the liquid flow in the first chamber (12) and the second chamber (13) similar, the energy of the two ultrasonic vibration waves can be maximized to form resonance on the target particles. By doing so, clogging of the filter membrane can be prevented, and separation efficiency can be further improved.

[0033] In one embodiment, two ultrasonic generators (30) are located on the same horizontal plane. That is, two ultrasonic vibration waves propagate in opposite directions and can overlap each other. Thus, clogging of the filter membrane can be prevented and separation efficiency can be further improved.

[0034] In one embodiment, within one cycle of turning on or off of the two ultrasonic generators (30), the turning ratio is 10% to 100%. Controlling the turning ratio of the two ultrasonic generators (30) can directly affect the magnitude of the total power output, and accordingly, can satisfy the power output required to detach different target particles (generally meaning target particles of different size or mass) from the filter membrane, thereby preventing the filter membrane from being clogged by different target particles and further improving filtration efficiency.

[0035] Referring again to FIGS. 4 to 6, the amplitude transformer (31) includes a first amplitude variation part (311), a second amplitude variation part (312), a third amplitude variation part (313), and a connecting part (314) that are sequentially connected. The connecting part (314) is connected to the piezoelectric ceramic assembly (32), and the first amplitude variation part (311) is coupled to the outer surface of the first chamber (12) or the outer surface of the second chamber (13) apart from the cross-section of the second amplitude variation part (312). In order to obtain an ultrasonic vibration wave that satisfies the aforementioned indicator, the size (h2) of the second amplitude variation part (312) is made smaller than the size (h1) of the first amplitude variation part (311) and the size (h3) of the third amplitude variation part (313) along a direction perpendicular to the extension direction of the amplitude transformer (31). In this embodiment, the first amplitude fluctuation section (311), the second amplitude fluctuation section (312), and the third amplitude fluctuation section (313) are all cylindrical structures, and the length of the second amplitude fluctuation section (312) is about 10 to 18 mm, preferably 15 mm, and the diameter is about 3 to 5 mm, preferably 4.5 mm; the length of the first amplitude fluctuation section (311) is about 4 to 7 mm, preferably 5 mm, and the diameter is about 5.5 to 6.5 mm, preferably 6 mm; and the length of the third amplitude fluctuation section (313) is about 13 to 17 mm, preferably 15 mm, and the diameter is about 5.5 to 6.5 mm, preferably 6 mm. FIGS. 7a to 7d show simulation photos (images) of the ultrasonic generator (30) generating ultrasonic vibration waves, and the amplitude transformer (31) of the aforementioned size can cause fine deformation in the horizontal direction, and the amount of deformation is about 0.04 nm.In this case, the ultrasonic vibration waves can achieve the purpose of disturbing the liquid sample inside the separation chip (10) by satisfying the aforementioned requirements, and can cause target particles attached to the first filter membrane (14) and the second filter membrane (15) to be effectively detached from the filter membrane.

[0036] Referring to FIGS. 5 and 6, the piezoelectric ceramic assembly (32) comprises a plurality of piezoelectric ceramic sheets (321) installed in a stacked manner, a plurality of electrode pieces (322) installed spaced apart from the plurality of piezoelectric ceramic sheets (321), an insulating sleeve (323), and a connecting member (324). The plurality of piezoelectric ceramic sheets (321) and the plurality of electrode pieces (322) are installed by covering the insulating sleeve (323), and the connecting member (324) penetrates the insulating sleeve (323) and is detachably connected to the amplitude transformer (31). By stacking and using multiple groups of piezoelectric ceramic sheets and electrode pieces (322), the frequency of the ultrasonic vibration waves is adjusted so that the frequency of the generated ultrasonic vibration waves is within the aforementioned range, thereby preventing clogging of the filter membrane and improving separation efficiency.

[0037] In one embodiment, the number of piezoelectric ceramic sheets (321) and electrode pieces (322) is each 4, and two electrode pieces (322) spaced apart from each other among the four electrode pieces (322) are each connected to the positive electrode of the power source, and the other two electrode pieces (322) are each connected to the negative electrode of the power source.

[0038] Referring again to FIGS. 5 and 6, the ultrasonic generator (30) further includes a control block (33) installed on the piezoelectric ceramic assembly (32) away from the amplitude transformer (31), and the connecting member (324) is also connected to the control block (33). The control block (33) primarily serves to balance and control the vibration frequency, and the weight of the control block (33) can be designed so that each ultrasonic generator (30) reaches the vibration frequency designed above.

[0039] Referring to FIG. 4, a first chamber (12) and a second chamber (13) are respectively installed on opposite sides of the sample pool (11) of the separation chip (10). The diameters of the first filter membrane (14) and the second filter membrane (15) may be the same or different from each other, and are determined according to the size of the target particles that actually need to be separated.

[0040] Referring to FIGS. 3 and FIGS. 8, the compression assembly (20) includes a base (21), a moving mechanism (22), and a driving mechanism (23). The moving mechanism (22) is installed on the driving shaft (231) of the driving mechanism (23). The driving shaft (231) of the driving mechanism (23) is rotated to drive the moving mechanism (22) to move along the driving shaft (231), thereby causing each of the ultrasonic generators (30) to move toward the separation chip (10) and be coupled to the outer surface of the first chamber (12) and the second chamber (13). Since the present invention can implement the interlocking of two ultrasonic generators (30) using a single compression assembly (20), unnecessary positioning mechanisms can be reduced and the structure of the equipment can be simplified.

[0041] The above movement mechanism (22) includes a first slide rail (221) installed on the base (21), two first slide blocks (222) installed on the first slide rail (221), a first mounting plate (223) installed on each of the first slide blocks (222), a second slide rail (224) installed on each of the first mounting plates (223), a second slide block (225) slidably installed on the second slide rail (224), a second mounting plate (226) installed on the second slide block (225), and an elastic member (227). One of the ultrasonic generators (30) is installed on one of the second mounting plates (226), and both ends of the elastic member (227) are connected to the first mounting plate (223) and the second mounting plate (226), respectively.

[0042] Each of the first mounting plates (223) is equipped with one ultrasonic generator (30). The driving mechanism (23) rotates the driving shaft (231) to cause the two first mounting plates (223) and the ultrasonic generators (30) installed thereon to move toward each other and come into contact with the surface of the separation chip (10). After contact, the ultrasonic generator (30) moves away from the separation chip (10) under the driving force of the separation chip (10), thereby compressing the elastic member (227). When the driving mechanism (23) is additionally driven to move the first mounting plate (223) toward the separation chip (10), the elastic force of the elastic member (227) propels the ultrasonic generator (30) to compress the separation chip (10). To separate the ultrasonic generator (30) from the separation chip (10), the driving shaft (231) is rotated in the opposite direction. By designing an elastic member (227) so that the ultrasonic generator (30) and the separation chip (10) are elastically connected, a rigid connection to the separation chip (10) can be prevented, and the risk of damage to the ultrasonic generator (30) to the separation chip (10) during the clamping process can be reduced.

[0043] Additionally, the elastic member (227) is a spring, and the clamping force of the ultrasonic generator (30) on the separation chip (10) is controlled by adjusting the hook elasticity coefficient of the elastic member (227) and the distance the ultrasonic generators (30) move toward each other. In this embodiment, if the distance the two ultrasonic generators (30) move toward each other is about 6 mm and the spring hook coefficient of the elastic member (227) is 1200 N / m, the clamping force on one side of the separation chip (10) is about 3.2 N.

[0044] Referring to FIG. 9, in another embodiment, it will be understood that the number of the first chamber (12) and the second chamber (13) in the separation chip (10a) may all be multiple, and the number of the first chamber (12) and the second chamber (13) can be designed according to actual needs. In addition, depending on the size and structural requirements of the actual separation chip (10a), multiple first chambers (12) or multiple second chambers (13) may be installed in parallel (as shown in FIG. 9) or installed side by side. The two adjacent first chambers (12) and the first chamber adjacent to the sample pool (11) and the sample pool (11) are connected through a single first filter membrane (14), and the diameter of each first filter membrane (14) may be designed differently to achieve the purpose of separating target particles of various sizes simultaneously. Likewise, the diameters of multiple second filter membranes (15) may also be different. Since the separation chip (10a) of multiple first chambers (12) and multiple second chambers (13) has a large number of filter membranes inside, filter membranes with small diameters are particularly prone to clogging. By installing multiple first filter membranes (14) and multiple second filter membranes (15) inside, the aforementioned parameters of the two ultrasonic generators (30) can be adjusted to reach a total output power requirement that prevents clogging of each filter membrane. In this way, filtration efficiency can be improved and purification quality can be guaranteed. Since there is no need to install a vibrating member on the filter membrane, the structure of the separation chip (10) can be simplified, and the difficulty and cost of manufacturing the separation chip (10a) can be reduced. In addition, since the above ultrasonic generator (30) is highly versatile, it can be applied to separation chips (10, 10a) of various structures or shapes, and by installing the ultrasonic generator (30) and the separation chip (10, 10a) separately, when ultrasonic separation is required, the outer surfaces of the ultrasonic generator (30) and the separation chip (10, 10a) can be combined, making operation simple.

[0045] Referring to FIGS. 1, 3, and 4, the differential pressure drive system (50) alternately generates a differential pressure in the first chamber (12) and the second chamber (13) of the separation chip (10). The differential pressure drive system (50) may be two independent differential pressure drive systems or a single differential pressure drive system designed. Here, a suitable differential pressure is selected according to the characteristics of the liquid sample, and the differential pressure of the differential pressure drive system (50) is selected within the range of -10 KPa to -50 KPa according to the liquid sample to be separated in the separation chip (10) of the present invention. That is, by alternately generating a differential pressure within the above range between the first chamber (12) or the second chamber (13) and the sample pool (11), the separation efficiency of the target particle material can be improved, and the filtration membrane can be effectively prevented from being clogged by particles. The differential pressure drive system (50) may include equipment such as a micro vacuum pump or a micro air pump. It will be understood that the differential pressure drive system (50) and the separation chip (10) can be connected via an adapter tube (53) with good airtightness. In one embodiment, the differential pressure drive system (50) includes a first vacuum pump (51) and a second vacuum pump (52), and the first vacuum pump (51) and the first opening (121) of the separation chip (10) are connected via one adapter tube (53), and the second vacuum pump (52) and the second opening (131) of the separation chip (10) are connected via another adapter tube (53).

[0046] In the process of implementing the connection between the differential pressure drive system (50) and the separation chip (10), two adapter tubes (53) may be installed in the compression assembly (20), and in the process of linking the compression assembly (20), the adapter tubes (53) and the ultrasonic generator (30) are simultaneously connected to the separation chip (10). In one embodiment, one of the adapter tubes (53) is installed on one of the first mounting plates (223) and is located below the corresponding ultrasonic generator (30). The drive mechanism (23) rotates the drive shaft (231) to cause the two adapter tubes (53) to move toward each other, so that the two adapter tubes (53) are respectively connected to the first opening (121) and the second opening (131) of the separation chip (10), and furthermore, the separation chip (10) is connected to the first vacuum pump (51) and the second vacuum pump (52), respectively.

[0047] In one embodiment, an elastic sleeve (54) is installed at one end of the adapter tube (53) that is close to the separation chip (10). In the process of connecting the separation chip (10) and the vacuum equipment, the adapter tube (53) must be connected to the opening of the separation chip (10) in a sealable manner; therefore, by adding the elastic sleeve (54), sealing at the opening side can be achieved after the adapter tube (53) and the separation chip (10) are connected. In this embodiment, the material of the elastic sleeve (54) is a variable soft material (e.g., rubber), so a sealing effect can be achieved by adjusting the compression amount of the elastic sleeve (54). The specific compression amount of the elastic sleeve (54) is determined by the Shore hardness of the elastic sleeve (54) and the driving force of the driving mechanism (23).

[0048] Referring to FIG. 3, the compression assembly (20) further includes a displacement sensing assembly (24), and the displacement sensing assembly (24) controls the pressure of the ultrasonic generator (30) and the adapter tube (53) on the separation chip (10) by measuring the distance traveled during the movement of the ultrasonic generator (30) and the adapter tube (53). The displacement sensing assembly (24) includes the first connecting piece (241), the second connecting piece (242), and the optical coupling member (243) installed on the first connecting piece (241), the first connecting piece (241) and the second connecting piece (242) are each fixed to two first mounting plates (223), and the optical coupling member (243) is slidably connected to one end of the second connecting piece (242) that is close to the first connecting piece (241). The above driving mechanism (23) also drives the first connecting piece (241) and the second connecting piece (242) to move along the driving shaft (231), and the optical coupling member (243) detects the relative distance of movement of the first connecting piece (241) and the second connecting piece (242). By installing the displacement sensing assembly (24) to detect the distance of movement of the ultrasonic generator (30) and the adapter tube (53), the connection between the ultrasonic generator (30) and the adapter tube (53) and the separation chip (10) can be better controlled, thereby enabling the realization of an optimal connection effect.

[0049] Referring to FIGS. 2 and FIGS. 4, the frequency conversion module (40) is electrically connected to the differential pressure drive system (50). The frequency conversion module (40) generates differential pressure alternately within the first chamber (12) and the second chamber (13) by controlling the power pressure provided to the differential pressure drive system (50). In one embodiment, the frequency conversion module (40) includes a frequency converter (41) and a control valve (42) connected to the frequency converter (41). The control valve (42) may be a liquid path converter, including but not limited to a solenoid valve or a rotary valve. The control valve (42) communicates with one of the first vacuum pump (51) and the second vacuum pump (52), respectively, thereby causing the first vacuum pump (51) and the second vacuum pump (52) to operate alternately and repeatedly. For example, the control valve (42) is connected to the first vacuum pump (51) so that the frequency converter (41) controls the operation of the first vacuum pump (51) and pumps air through the first opening (121) to generate a differential pressure within the first chamber (12), and causes the liquid in the liquid sample in the sample pool (11) and the component whose size is smaller than the diameter of the first filter membrane (14) to enter the first chamber (12) through the first filter membrane (14) under the action of the differential pressure. Next, the frequency converter (41) controls the first vacuum pump (51) to stop operating. Next, the control valve (42) is switched to communicate with the second vacuum pump (52) so that the frequency converter (41) controls the operation of the second vacuum pump (52), and air is pumped through the second opening (131) to generate a differential pressure within the second chamber (13), and the liquid in the liquid sample in the sample pool (11) and the component whose size is smaller than the diameter of the second filter membrane (15) are allowed to enter the second chamber (13) through the second filter membrane (15) under the action of the differential pressure. Then, the frequency converter (41) controls the second vacuum pump (52) to stop operating. The above steps are repeated several times.

[0050] The controller (60) controls the vibration of two ultrasonic generators (30) to generate two ultrasonic vibration waves when the first chamber (12) stops pumping. The controller (60) also controls the vibration of two ultrasonic generators (30) to generate two ultrasonic vibration waves when the second chamber (13) stops pumping. Here, the controller (60) can be electrically connected to the first vacuum pump (51) and the second vacuum pump (52), and when the first vacuum pump (51) or the second vacuum pump (52) stops operating, the controller (60) determines that the first vacuum pump (51) has stopped pumping in the first chamber (12) or that the second vacuum pump (52) has stopped pumping in the second chamber (13), and at this time, the controller can notify the two ultrasonic generators (30) to start vibrating.

[0051] An embodiment of the present invention further provides a method for separating target particles from a liquid sample applied to the separation chip (10), and the method comprises the following steps.

[0052] In step S1, the separation chip (10) of the present invention is provided, and a liquid sample is provided to the sample pool (11) of the separation chip (10).

[0053] In step S2, the compression assembly (20) is controlled to drive the two ultrasonic generators (30) toward the separation chip (10), so that the two ultrasonic generators (30) are connected to the outer surface of the outermost first chamber (12) and the outer surface of the outermost second chamber (13), respectively, in close proximity to the cross-section of the separation chip (10).

[0054] At the same time as the ultrasonic generator (30) is connected, the compression assembly (20) also drives two adapter tubes (53) to communicate with the first opening (121) and the second opening (131), respectively, thereby allowing the first opening (121) and the second opening (131) to communicate with the differential pressure drive system (50), respectively.

[0055] In step S3, the first chamber (12) is pumped through the first opening (121) to generate a differential pressure within the first chamber (12).

[0056] The differential pressure drive system (50) pumps the at least one first chamber (12) through the first opening (121) to generate a differential pressure within the at least one first chamber (12). The liquid and components of the liquid sample (11) in the sample pool (11) and components that are smaller in size than the diameter of the first filter membrane (14) move toward the first filter membrane (14) under the action of the differential pressure and enter the at least one first chamber (12) through the first filter membrane (14).

[0057] In step S4, the pumping of the at least one first chamber (12) is stopped, and the vibration of the two ultrasonic generators (30) is controlled to generate ultrasonic vibration waves. At the same time, the second chamber (13) is pumped through the second opening (131) to generate a differential pressure within the at least one second chamber (13).

[0058] Here, the ultrasonic vibration wave causes the liquid sample, the first filter membrane (14), and the second filter membrane (15) to vibrate at high frequency, thereby allowing target particles adsorbed in the pores of the filter membrane to be rapidly separated from the pores of the filter membrane and resuspended in the refluxed liquid sample, and preventing the target particles from aggregating. At the same time, the differential pressure driving system (50) pumps the second chamber (13) through the second opening (131) to generate a differential pressure within the second chamber (13). Components attached to the surface of the first filter membrane (14) may flow into the sample pool (11) according to the gas flow and / or liquid flow, and components of the liquid sample in the sample pool (11) and components whose size is smaller than the diameter of the second filter membrane (15) move toward the second filter membrane (15) under the action of the differential pressure and enter the at least one second chamber (13) through the second filter membrane (15).

[0059] In step S5, the pumping of at least one second chamber (13) is stopped, and the vibration of the two ultrasonic generators (30) is controlled to generate ultrasonic vibration waves.

[0060] Next, steps S3 to S5 can be repeated several times, so that components smaller than the diameter of the filter membrane in the liquid sample are removed and components larger than the diameter of the filter membrane remain in the sample pool (11), thereby achieving a superior separation and purification effect.

[0061] Compared to the prior art, the separation device (100) of the present invention separately installs a separation chip (10) and an ultrasonic generator (30), and when in use, moves two ultrasonic generators (30) through the linkage of a compression assembly (20) to connect to the separation chip (10), and provides ultrasonic vibration waves to the separation chip (10) to vibrate the separation chip (10), thereby preventing clogging of the first filter membrane (14) and the second filter membrane (15), and improving separation efficiency and separation purity; and since there is no need to install a vibration member on the separation chip (10), the difficulty and cost of manufacturing the separation chip (10) can be reduced.

[0062] The ultrasonic generator (30) has a specific structure and parameters that match the separation chip (10), and the generated ultrasonic vibration waves can ensure that particles attached to the first filter membrane (14) and the second filter membrane (15) are removed, thereby preventing clogging of the filter membrane and achieving the purpose of improving filtration efficiency and filtration purity.

[0063] The compression assembly (20) implements the interlocking of two ultrasonic generators (30) so that the two ultrasonic generators (30) are each connected to or separated from the separation chip (10), and the interlocking process is simple to operate and the ultrasonic generators (30) and the separation chip (10) are closely connected, so that ultrasonic vibration waves can be transmitted to the first filter membrane (14) and the second filter membrane (15) without damaging the separation chip (10).

[0064] By installing two adapter tubes (53) in the compression assembly (20), the connection of the ultrasonic generator (30), the differential pressure drive system (50), and the separation chip (10) can be simultaneously implemented during the interlocking process, the connection structure can be simplified, and the complexity of the structure and the difficulty of operation can be reduced. Explanation of the symbols

[0065] 100: Separation device 10, 10': Separation chip 11, 11': Sample pool 12, 12': First chamber 121: First opening 13, 13': Second chamber 131: Second opening 14, 14': First filtration membrane 15, 15': Second filtration membrane 16': Vibration member 70: Vibration System 20: Compression Assembly 21: Bass 22: Movement Mechanism 221: 1st slide rail 222: 1st Slide Block 223: 1st mounting plate 224: Second slide rail 225: 2nd slide block 226: Second mounting plate 227: Elastic member 23: Driving Mechanism 231: Drive shaft 24: Displacement sensing assembly 241: First Connection 242: Second Connection 243: Optical coupling member 30: Ultrasonic generator 31: Amplitude Transformers 311: First amplitude variation section 312: Second amplitude variation section 313: Third Amplitude Variation Section 314: Connection 32: Piezoelectric ceramic assembly 321: Piezoelectric ceramic sheet 322: Electrode 323: Insulation Sleeve 324: Connecting member 33: Control Block 40: Frequency conversion module 41: Frequency converter 42: Control valve 50: Differential pressure drive system 51: First vacuum pump 52: Second vacuum pump 53: Adapter tube 54: Elastic Sleeve 60: Controller h1, h2, h3: Size

Claims

Claim 1 A separation device for separating and purifying target particles from a liquid sample, wherein the separation device comprises a sample pool, at least one first chamber located on one side of the sample pool, and at least one second chamber located on the other side of the sample pool away from the at least one first chamber, wherein the two adjacent first chambers and the first chamber adjacent to the sample pool and the sample pool are all connected through a first filter membrane, and the two adjacent second chambers and the second chamber adjacent to the sample pool and the sample pool are all connected through a second filter membrane, and wherein the diameter of each of the first filter membrane and each of the second filter membranes is smaller than the particle diameter of the target particles; a separation chip comprising a compression assembly and two ultrasonic generators located in the compression assembly, wherein the compression assembly drives the ultrasonic generators to move toward the separation chip, thereby causing the two ultrasonic generators to be connected to the outer surface of the outermost first chamber and the outer surface of the outermost second chamber, respectively, in close proximity to the cross-section of the separation chip. A separation device comprising: a system; a differential pressure driving system; a frequency conversion module each connected to the first chamber and the second chamber through the differential pressure driving system and controlling the differential pressure driving system to alternately generate differential pressure in the first chamber and the second chamber; and a controller that controls the vibration of two ultrasonic generators to generate ultrasonic vibration waves each when the differential pressure driving system stops generating differential pressure in the first chamber or the second chamber, wherein the vibration frequencies of the two ultrasonic vibration waves are all 15 KHz to 80 KHz; and within one cycle of the ultrasonic vibration waves, the duty ratio of the two ultrasonic vibration waves is all 10% to 90%. Claim 2 delete Claim 3 A separation device according to claim 1, characterized in that the difference between the vibration frequency and the duty ratio of the two ultrasonic vibration waves is 30% or less. Claim 4 A separation device according to claim 1, characterized in that, within one cycle of turning on or off of the two ultrasonic generators, the ratio of turning on is 10% to 100%. Claim 5 A separation device according to claim 1, characterized in that the two ultrasonic generators are located on the same horizontal plane. Claim 6 A separation device according to claim 1, characterized in that the ultrasonic generator comprises an amplitude transformer installed in close proximity to the separation chip and a piezoelectric ceramic assembly connected to the amplitude transformer. Claim 7 A separation device according to claim 6, wherein the amplitude transformer comprises a first amplitude variation part, a second amplitude variation part, a third amplitude variation part and a connecting part connected sequentially, and the connecting part is connected to the piezoelectric ceramic assembly, and the first amplitude variation part is coupled to the outer surface of the first chamber or the outer surface of the second chamber apart from the cross-section of the second amplitude variation part, and the size of the second amplitude variation part is smaller than the size of the first amplitude variation part and the third amplitude variation part along a direction perpendicular to the extension direction of the amplitude transformer. Claim 8 A separation device according to claim 6, wherein the piezoelectric ceramic assembly comprises a plurality of piezoelectric ceramic sheets installed in a stacked manner, a plurality of electrode pieces installed spaced apart from the plurality of piezoelectric ceramic sheets, an insulating sleeve, and a connecting member, wherein the plurality of piezoelectric ceramic sheets and the plurality of electrode pieces are installed by covering the insulating sleeve, and the connecting member penetrates the insulating sleeve and is detachably connected to the amplitude transformer. Claim 9 A separation device according to claim 8, wherein the ultrasonic generator further comprises a control block, the control block is installed at one end of the piezoelectric ceramic assembly separated from the amplitude transformer, and the connecting member is connected to the control block. Claim 10 A separation method for separating and purifying target particles from a liquid sample, the method comprising: providing a separation device according to any one of claims 1, 3 to 9, and providing a liquid sample to the sample pool; controlling the compression assembly to drive two ultrasonic generators to move toward the separation chip so that two ultrasonic generators are respectively coupled to the outer surface of the outermost first chamber and the outer surface of the outermost second chamber in close proximity to the cross-section of the separation chip; generating a differential pressure in the at least one first chamber so that a component of the liquid sample in the sample pool whose particle size is smaller than the diameter of the first filter membrane enters the at least one first chamber under the action of the differential pressure; stopping the generation of the differential pressure in the at least one first chamber and controlling the vibration of the two ultrasonic generators to generate an ultrasonic vibration wave; generating a differential pressure in the at least one second chamber so that a component of the liquid sample in the sample pool whose particle size is smaller than the diameter of the second filter membrane enters the at least one second chamber under the action of the differential pressure. A separation method characterized by comprising the steps of: stopping the generation of differential pressure in at least one second chamber and controlling the vibration of two ultrasonic generators to generate ultrasonic vibration waves.

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