Droplet generating device

Abstract

Disclosed is a droplet generating device for generating droplets using a prepared PCR reaction solution and oil. In particular, the droplet generating device comprises: a pump providing negative pressure; a movable rigid body having a cubic shaped body and including a plurality of pressure applying ports and at least one pump connecting port; a flexible tube connecting the pump and the at least one pump connecting port; a motor providing a mechanical force; and a mechanical mechanism for moving the movable rigid body along a first axis by using the mechanical force provided from the motor.

Description

droplet generating device

[0001] The present disclosure relates to a droplet generation device. More specifically, the disclosure relates to a droplet generation device capable of stably applying pressure to a droplet generation cartridge into which a PCR reaction solution and oil are dispensed to generate droplets.

[0002] Polymerase Chain Reaction (PCR) is a molecular biological technique that replicates and amplifies a desired portion of extracted DNA. PCR allows researchers to selectively amplify only specific DNA fragments of interest from solutions containing extremely complex and minute quantities of DNA, such as the human genome. Furthermore, PCR requires a short amplification time, is simple to perform, and can be performed using fully automated equipment. Consequently, PCR is widely used in a wide range of DNA-related fields, including molecular biology, medicine, forensic investigation, and biological classification.

[0003] Meanwhile, PCR can be divided into the first generation PCR, which qualitatively analyzes genes such as collected DNA through electrophoresis, the second generation real-time PCR, which can perform both qualitative and quantitative analysis of genes, and digital PCR (dPCR), which creates genes as tens of thousands of droplets, detects the fluorescence of the droplets, and can perform qualitative and quantitative analysis of genes.

[0004] Among these, digital PCR detects fluorescence from each amplified droplet and counts the number of positive and negative droplets, thereby allowing analysis of the status of the collected genes, the onset of a disease, or infection.

[0005] Meanwhile, accurately counting the number of positive and negative droplets in digital PCR is crucial. However, if the droplets produced are not uniform in size, the number of samples or fluorescent substances contained in each droplet may vary. If the number of fluorescent substances or samples in each droplet varies, the intensity of fluorescence emitted by each positive droplet may vary. This can reduce the reliability of distinguishing between positive and negative droplets.

[0006] Therefore, in order to accurately determine the number of positive and negative droplets, it is important that the size of the droplets created during droplet generation is as consistent as possible.

[0007] The present disclosure relates to a droplet generating device.

[0008] The problem to be solved by the present disclosure is to provide a droplet generation device capable of providing a stable pressure to a droplet generation cartridge so that the size of the droplets generated by mixing a PCR reaction solution and oil is constant.

[0009] The problems to be solved by the present disclosure are not limited to the problems described above, and problems not mentioned can be clearly understood by a person having ordinary skill in the technical field corresponding to the present disclosure from this specification and the attached drawings.

[0010] A droplet generating device for generating droplets using a prepared PCR reaction solution and oil according to the present disclosure may include: a pump for providing negative pressure; a movable rigid body having a cubic shaped body and including a plurality of pressure applying ports and at least one pump connecting port, wherein the plurality of pressure applying ports and the at least one pump connecting port are formed on a surface of the movable rigid body, and the movable rigid body further includes an airway network formed inside the movable rigid body and connecting the plurality of pressure applying ports and the at least one pump connecting port; a flexible tube for connecting the pump and the at least one pump connecting port; a motor for providing mechanical force; and a mechanical mechanism for moving the movable rigid body along a first axis using the mechanical force provided from the motor.

[0011] At this time, the flexibility of the flexible tube can maintain a fluid connection between the pump and the at least one pump connection port regardless of the position of the movable rigid body, and the rigidity of the air passage network can enable the negative pressure provided by the pump to be stably applied to a plurality of droplet generation reservoirs included in the droplet generation cartridge.

[0012] According to one embodiment of the present disclosure, the droplet generating device can generate droplets with a uniform size.

[0013] The effects according to the present disclosure are not limited to the effects described above, and effects not mentioned can be clearly understood by a person having ordinary skill in the art to which the present invention pertains from the present disclosure and the attached drawings.

[0014] Figure 1 illustrates a droplet generator cartridge used in an embodiment of the present disclosure.

[0015] Figure 2 is intended to explain the problems of conventional droplet generating devices.

[0016] FIGS. 3 to 5 are drawings for explaining a droplet generating device according to a first embodiment of the present disclosure.

[0017] FIGS. 6 and 7 are drawings for explaining a droplet generating device according to a second embodiment of the present disclosure.

[0018] FIGS. 8 and 9 are drawings for explaining a droplet generating device according to a third embodiment of the present disclosure.

[0019] FIGS. 10 to 13 are drawings for explaining components included in a droplet generating device according to an embodiment of the present disclosure.

[0020] FIGS. 14 and 15 are for explaining the operation of a droplet generating device according to an embodiment of the present disclosure.

[0021] Figures 16 and 17 illustrate an experimental design for comparing droplet generation results in embodiments of the present disclosure and a conventional droplet generation device.

[0022] Figures 18 to 30 show experimental results of generating droplets using embodiments of the present disclosure and a conventional droplet generating device.

[0023] The above-described purposes, features, and advantages of the present disclosure will become more apparent through the following detailed description taken in conjunction with the accompanying drawings. However, since the present disclosure is susceptible to various modifications and various embodiments, specific embodiments will be illustrated in the drawings and described in detail below.

[0024] Since the embodiments described in this specification are intended to clearly explain the spirit of the present disclosure to a person having ordinary skill in the art to which the present disclosure pertains, the present disclosure is not limited to the embodiments described in this specification, and the scope of the present disclosure should be interpreted to include modified or altered examples that do not depart from the spirit of the present disclosure.

[0025] The drawings attached to this specification are intended to facilitate explanation of the present disclosure, and the shapes depicted in the drawings may be exaggerated as necessary to help understanding of the present disclosure, and thus the present disclosure is not limited by the drawings.

[0026] If a detailed description of a known function or configuration related to this disclosure is deemed to unnecessarily obscure the gist of this disclosure, such detailed description will be omitted. Furthermore, numbers (e.g., "first," "second," etc.) used throughout the description of this specification are merely identifiers used to distinguish one component from another.

[0027] In addition, the suffixes "unit," "module," and "part" used for components in the description below are given or used interchangeably only for the convenience of writing the specification, and do not have distinct meanings or roles in themselves.

[0028]

[0029] A droplet generating device for generating droplets using a prepared PCR reaction solution and oil according to the present disclosure may include: a pump providing negative pressure; a movable rigid body having a cubic shaped body and including a plurality of pressure applying ports and at least one pump connecting port; a flexible tube connecting the pump and the at least one pump connecting port; a motor providing mechanical force; and a mechanical mechanism for moving the movable rigid body along a first axis using the mechanical force provided from the motor.

[0030] Here, the plurality of pressure application ports and the at least one pump connection port are formed on a surface of the movable rigid body, and the movable rigid body may further include an airway network formed inside the movable rigid body to connect the plurality of pressure application ports and the at least one pump connection port.

[0031] Additionally, the flexibility of the flexible tube allows a fluid connection between the pump and the at least one pump connection port to be maintained regardless of the position of the movable rigid body, and the rigidity of the air passage network allows a negative pressure generated by the pump to be stably applied to a plurality of droplet generation reservoirs included in the droplet generation cartridge.

[0032] Meanwhile, the droplet generation cartridge includes a plurality of PCR reaction solution reservoirs into which a PCR reaction solution is dispensed, a plurality of oil reservoirs into which oil is dispensed, and the plurality of droplet generation reservoirs, and a negative pressure stably applied to the plurality of droplet generation reservoirs can cause the PCR reaction solutions discharged from the plurality of PCR reaction solution reservoirs and the oils discharged from the plurality of oil reservoirs to meet and generate droplets, and the sizes of the droplets stored in the plurality of droplet generation reservoirs can be constant.

[0033] Additionally, the plurality of pressure application ports may be positioned on the lower surface of the movable rigid body to apply negative pressure to the plurality of droplet generation reservoirs.

[0034] Additionally, the first axis may be perpendicular to the surface on which the plurality of pressure application ports are formed.

[0035] In addition, the movable rigid body may further include a valve that allows negative pressure provided from the pump to be applied to the plurality of droplet generation reservoirs; and the air passage network may include an air conduit network connecting the valve and the plurality of pressure application ports and an air conduit connecting the valve and the at least one pump connection port.

[0036] In addition, the droplet generation device may further include a controller that controls the operation of the pump and the valve, and the controller may control the pump so that the negative pressure is started to be provided at a first time point, and open the valve so that the negative pressure provided from the pump is applied to the plurality of droplet generation reservoirs after a first time interval from the first time point. At this time, the first time interval may be preset.

[0037] In addition, the droplet generating device may further include a position sensor for sensing a position of the movable rigid body; and a controller for controlling the motor; wherein the controller controls the motor based on sensing of the position sensor so that the plurality of pressure application ports can be fluidly connected to the plurality of droplet generating reservoirs, thereby causing the mechanical mechanism to move the movable rigid body downward along the first axis.

[0038] Additionally, the controller may control the motor based on sensing of the position sensor to cause the mechanical mechanism to move the movable rigid body upward along the first axis such that the plurality of pressure application ports cannot be fluidly connected to the plurality of droplet generation reservoirs.

[0039]

[0040] [Digital PCR General]

[0041] Digital PCR boasts approximately 1,000 times higher sensitivity than real-time PCR, facilitating the analysis of target genes even in complex mixtures. The high sensitivity of digital PCR makes it more useful than real-time PCR in cases where the target concentration in the sample is very low.

[0042] Additionally, digital PCR is very advantageous in quantifying target DNA compared to real-time PCR.

[0043] To perform digital PCR, a process is fundamentally required: dividing a DNA-containing sample (solution) into predetermined volumes, and inducing an amplification reaction of the target DNA within the divided sample (solution). Specifically, digital PCR methods can be categorized in various ways depending on the method of dividing the sample into predetermined volumes. The present disclosure relates to droplet digital PCR, which utilizes a method of dividing the sample into droplets.

[0044]

[0045] To perform droplet digital PCR, there are three main steps:

[0046]

[0047] (1) Droplet generation stage

[0048] (a) A sample containing target DNA to be amplified and (b) a PCR reaction solution containing i) primers for amplifying the target DNA, ii) a fluorescent probe or fluorescent dye, iii) dNTP (deoxynucleoside triphosphate) used as a material for an amplicon, iv) a polymerase, etc. are prepared. For example, the fluorescent probe or fluorescent dye may be FAM, HEX, VIC, TAMRA, EvaGreen, etc. In addition, the sample refers to the result of pretreatment necessary for PCR analysis, such as blood, tissue, cells, saliva, etc. collected from a subject. In particular, the sample contains DNA to be analyzed.

[0049]

[0050] Fig. 1 illustrates a droplet generation cartridge (100) for generating droplets using a PCR reaction solution and oil. Fig. 1(a) illustrates a top view of the droplet generation cartridge (100), and Fig. 1(b) illustrates a bottom view of the droplet generation cartridge (100).

[0051] Referring to Fig. 1(a), the droplet generating cartridge (100) may include at least one droplet generating unit (110). The droplet generating cartridge (100) may include a plurality of droplet generating units (110).

[0052] The droplet generation unit (110) includes an oil reservoir (111), a PCR reaction solution reservoir (112), and a droplet reservoir (113). Oil is dispensed into at least one oil reservoir (111) included in the droplet generation cartridge, and PCR reaction solution is dispensed into at least one PCR reaction solution reservoir (112) included in the droplet generation cartridge.

[0053] Meanwhile, when negative pressure is applied to the droplet reservoir (113), the oil stored in the oil reservoir (111) and the PCR reaction liquid stored in the PCR reaction liquid reservoir (112) move to the droplet reservoir (113), thereby generating droplets.

[0054] Alternatively, when positive pressure is applied to the oil reservoir (111) and the PCR reaction liquid reservoir (112), the oil stored in the oil reservoir (111) and the PCR reaction liquid stored in the PCR reaction liquid reservoir (112) move to the droplet reservoir (113), thereby generating droplets.

[0055] Figure 1(b) is a bottom view of a droplet generating cartridge (100) for explaining in detail the process of generating droplets in a droplet generating unit (110).

[0056] Referring to FIG. 1(b), the droplet generation unit (110) further includes a droplet generation junction (130), a plurality of oil transfer channels (121) extending between an oil reservoir (111) and the droplet generation junction (130), a PCR reaction solution transfer channel (122) extending between a PCR reaction solution reservoir (112) and the droplet generation junction (130), and a droplet transfer channel (123) extending between the droplet reservoir (113) and the droplet generation junction (130). That is, the plurality of oil transfer channels (121), the PCR reaction solution transfer channels (122), and the droplet transfer channels (123) join at the droplet generation junction (130).

[0057] When negative pressure is applied to the droplet reservoir (113) or positive pressure is applied to the oil reservoir (111) and the PCR reaction solution reservoir (122), the oil stored in the oil reservoir (111) moves to the droplet generation junction (130) through the plurality of oil movement channels (121), and the PCR reaction solution stored in the PCR reaction solution reservoir (122) moves to the droplet generation junction (130) through the PCR reaction solution transfer channel (122). The PCR reaction solution and the oil that meet at the droplet generation junction (130) are combined so that the oil surrounds the PCR reaction solution, thereby generating droplets. The generated droplets are then transferred to the droplet reservoir (113).

[0058] That is, the force by which the PCR reaction solution and oil are transferred and meet at the liquid crystal generation junction (130) and transferred to the droplet reservoir (113) is generated by the negative pressure applied to the droplet reservoir (113) or the positive pressure applied to the oil reservoir (111) and the PCR reaction solution reservoir (122), and no other force is applied.

[0059] Meanwhile, the method of generating droplets using PCR reaction solution and oil is based on microfluidics technology, but is not limited thereto.

[0060]

[0061] (2) Droplet amplification stage

[0062] As described above, after the droplets are generated, they are repeatedly heat-treated according to a predetermined thermal cycle. This allows the target DNA to be amplified within the PCR reaction solution contained within each droplet. While DNA amplification occurs within droplets containing target DNA, it does not occur within droplets that do not contain target DNA. Basically, the positive droplets described above refer to droplets containing target DNA, and the negative droplets refer to droplets that do not contain target DNA.

[0063]

[0064] (3) Droplet detection step

[0065] Meanwhile, in general, when two or more target DNAs are to be detected using two or more fluorescent substances, PCR reagents can be designed so that each different fluorescence corresponds to a different target DNA. That is, when a first target DNA is amplified, fluorescence emitted from the amplified amplicon is formed in a first wavelength band, and when a second target DNA is amplified, fluorescence emitted from the amplified amplicon is formed in a second wavelength band that is distinct from the first wavelength band. Accordingly, a PCR device having a plurality of fluorescence detectors can detect a plurality of target DNAs from a single sample.

[0066] In digital PCR, since the PCR reaction occurs within a single droplet, detecting the type (wavelength) of fluorescence emitted from a completed amplification droplet provides information about the target DNA contained within that droplet. Therefore, in digital PCR equipment, it is crucial to accurately detect the type of fluorescence emitted from each droplet.

[0067] Fluorescence is detected from droplets containing amplified target DNA (positive droplets) using a fluorescence detector. That is, the fluorescence detector outputs light (e.g., a laser or LED) toward the droplets and can detect the wavelength of fluorescence emitted when the output light interacts with the fluorescent material contained in the droplets.

[0068] At this time, the device including the fluorescence detector can count the droplets for each fluorescent substance according to the wavelength of fluorescence corresponding to the fluorescent substance. For example, the device can individually count the number of droplets having FAM, the number of droplets having HEX, the number of droplets having VIC, the number of droplets having TAMRA, and the number of droplets having EvaGreen among the amplified droplets according to the wavelength detected by the fluorescence detector. For example, the detector of the fluorescence detector can detect only a specific wavelength. In other words, the detector can include a filter that passes only a specific wavelength, and thus, can detect only the wavelength emitted by a specific fluorescent substance.

[0069] Accordingly, each of the plurality of fluorescence detectors can detect a distinct wavelength, and based on the number of times each detector detects its corresponding wavelength, the device can individually count the number of droplets among the amplified droplets that have a fluorescent substance corresponding to the wavelength that each detector can detect.

[0070] As a specific example, when the intensity of fluorescence detected by the fluorescence detector exceeds a certain threshold, the number of counted droplets for the fluorescent substance corresponding to the wavelength that the fluorescence detector can detect can be increased.

[0071] The device can analyze biological properties of a sample based on the number of droplets per counted fluorescent substance.

[0072]

[0073] Meanwhile, the aforementioned droplet generation step, droplet amplification step, and droplet detection step may be performed separately in different devices. For example, the droplet generation step may be performed by a first device, the droplet amplification step may be performed by a second device, and the droplet detection step may be performed by a third device.

[0074] As another example, the above-described droplet generation step, droplet amplification step, and droplet detection step may be performed integratedly in one specific device.

[0075] As another example, two of the above-described droplet generation steps, droplet amplification steps, and droplet detection steps may be performed in one device, and the remaining one may be performed in another device. For example, the droplet generation step and the droplet amplification step may be performed in a fourth device, and the droplet detection step may be performed in the third device. As another example, the droplet generation step may be performed in the first device, and the droplet amplification step and the droplet detection step may be performed in a fifth device.

[0076]

[0077] Meanwhile, the droplet generating device described in the present disclosure is a device that performs a droplet generating step. However, this droplet generating device does not have to perform only the droplet generating step. That is, the droplet generating device may be the first device described above, a specific device, or a fourth device. Therefore, the droplet generating device may be a device that includes the components described in the present disclosure, which will be described later, and can perform the functions and operations. In other words, the droplet generating device of the present disclosure may include at least one other component other than the components described in the present disclosure, and may also perform other functions and operations other than the functions and operations described in the present disclosure.

[0078]

[0079] [Droplet generator]

[0080] As described above, in order for droplets to be generated in the droplet generation cartridge (100), pressure must be provided to the droplet generation cartridge (100) so that the PCR reaction solution stored in the PCR reaction solution reservoir (122) and the oil stored in the oil reservoir (111) can move to the droplet reservoir (113).

[0081] The droplet generating device described in the present disclosure is a device that functions to apply the pressure to the droplet generating cartridge so that droplets can be generated in the droplet generating cartridge (100).

[0082] For example, the droplet generating device is a device that applies pressure generated by a pump to a droplet generating cartridge (100) through a plurality of pressure application ports, thereby allowing droplets to be generated in a droplet reservoir (113) of the droplet generating cartridge (100).

[0083] As described above, the pressure applied to the droplet generating cartridge (100) may be a negative pressure applied to the droplet reservoir (113) or a positive pressure applied to the oil reservoir (111) and the PCR reaction solution reservoir (122). However, the following description assumes that the droplet generating cartridge (100) applies a negative pressure to the droplet reservoir (113). However, the following description does not mean to exclude an embodiment in which the droplet generating device applies a positive pressure provided to the oil reservoir (111) and the PCR reaction solution reservoir (122), but rather, it is clear that the subject matter described by the present disclosure can be applied to an embodiment in which the droplet generating device applies a positive pressure provided to the oil reservoir (111) and the PCR reaction solution reservoir (122).

[0084]

[0085] [Problems with conventional droplet generation devices]

[0086] Figure 2 shows a method for generating droplets in a conventional droplet generating device.

[0087] Referring to FIG. 2, the droplet generating device includes a pump (200) and a pressure applying structure. Here, the pressure applying structure includes a plurality of pressure applying ports (310) and a flexible tube. In addition, although not shown in FIG. 2, the droplet generating device may further include a motor. The droplet generating device may control one or more motors connected to the pressure applying structure to move the plurality of pressure applying ports (310) and / or the droplet generating cartridge (100).

[0088] In the following description, the meaning that the plurality of pressure application ports (310) and / or the droplet generation cartridge (100) move may mean that the droplet generation device controls one or more motors to move the plurality of pressure application ports (310) and / or the droplet generation cartridge (100). In addition, the meaning that the pump (200) generates pressure may mean that the droplet generation device controls the pump (200) to generate pressure.

[0089] The flexible tube transmits the pressure generated by the pump (200) to a plurality of pressure application ports (310). In addition, the plurality of pressure application ports (310) apply the transmitted pressure to the droplet generation cartridge (100). For example, the plurality of pressure application ports (310) may apply negative pressure to the droplet reservoir (113) or positive pressure to the oil reservoir (111) and the PCR reaction solution reservoir (1120).

[0090] Referring to Fig. 2 (a), the droplet generating cartridge (100) moves inside the droplet generating device. For example, the droplet generating cartridge (100) moves inside the droplet generating device along the direction (1).

[0091] Additionally, the plurality of pressure application ports (310) move to a first position on the top of the droplet generation cartridge (100). At this time, the flexible tube moves according to the movement of the plurality of pressure application ports (310). For example, the flexible tube may be deformed (e.g., the flexible tube gradually straightens) until the plurality of pressure application ports (310) move in the direction (2) and are positioned at the first position on the top of the droplet generation cartridge (100).

[0092] Referring to FIG. 2 (b), the plurality of pressure application ports (310) move downward to a second position and are fluidly connected to the droplet generation cartridge (100). At this time, the flexible tube moves according to the movement of the plurality of pressure application ports (310). For example, the flexible tube may move until the plurality of pressure application ports (310) move to a second position in which they are fluidly connected to the droplet generation cartridge (100) along the direction (3).

[0093] As described above, the plurality of pressure application ports (310) can move to a second position to be fluidly connected with the droplet generation cartridge (100). In addition, when droplet generation is completed, the plurality of pressure application ports (310) can move in the opposite direction of direction (3) to a third position and then move in the opposite direction of direction (2) to a fourth position to release the fluidic connection with the droplet generation cartridge (100).

[0094] When the plurality of pressure application ports (310) are moved to be fluidly connected to or disengaged from the droplet generation cartridge (100), the flexible tube facilitates smooth movement of the plurality of pressure application ports (310).

[0095] In other words, implementing a pathway network through which pressure is transmitted using a flexible tube is for smooth movement of multiple pressure application ports (310).

[0096]

[0097] Meanwhile, when a conventional droplet generating device generates droplets according to the above-described process, a phenomenon occurs in which the droplets are not generated uniformly in size. In particular, when vibration occurs in the droplet generating device due to a force provided from outside the droplet generating device, or when vibration occurs within the droplet generating device due to components included in the droplet generating device (e.g., a motor or a pump), a phenomenon in which the droplets are not generated uniformly in size occurs, and it is found that the greater the generated vibration, the greater the size difference between the droplets.

[0098] Non-uniformity in size between droplets may mean that the pressure applied to the droplet generating cartridge (100) is not stable.

[0099] Here, stable pressure means that the pressure remains constant during the time period during which pressure is applied to the droplet generating cartridge (100).

[0100] Here, the pressure being maintained constant means that the change in pressure during the time period in which pressure is applied to the droplet generating cartridge (100) changes within a first predetermined range and / or that the amount of change in pressure per unit time changes within a second predetermined range.

[0101] On the other hand, the pressure being unstable (i.e., the pressure is not maintained constant) means that the pressure applied to the droplet generating cartridge (100) changes over a first predetermined range during the time period and / or the amount of change in pressure per unit time changes over a second predetermined range.

[0102] When droplets are generated, if their size is not controlled to ensure uniformity, the number of samples or fluorescent substances contained in each droplet may vary. Furthermore, if the number of fluorescent substances or samples in each droplet varies, the intensity of the fluorescence emitted by each positive droplet may vary. This leads to unreliable digital PCR analysis results.

[0103] Accordingly, a structure of a droplet generation device capable of controlling the size of droplets to be uniform is required. In other words, a structure of a droplet generation device capable of controlling the pressure applied to a liquid crystal generation cartridge (100) to be stable is required.

[0104]

[0105] To address the above-described problems, the present disclosure proposes a method for implementing a pressure-transmitting pathway network (hereinafter, referred to as a pressure applying pathway network) between a plurality of pressure applying ports and a pump so that at least a portion thereof has rigidity. In other words, the present disclosure proposes a method for implementing a pressure-transmitting pathway network implemented as a flexible tube in FIG. 2 so that at least a portion thereof has rigidity.

[0106] In addition, through this, we intend to propose a method in which the pressure generated when the pump (200) generates pressure is stably applied to the droplet generating cartridge (100).

[0107]

[0108] Meanwhile, in the description described above and the description described below, the fact that the plurality of pressure application ports (310) and the droplet generation cartridge (100) are fluidly connected may mean that the plurality of pressure application ports (310) and the droplet reservoir (113) are fluidly connected. As described above, the plurality of pressure application ports (310) and the droplet reservoir (113) are fluidly connected when the pump (200) generates negative pressure.

[0109] On the other hand, the fact that the plurality of pressure application ports (310) and the droplet generation cartridge (100) are fluidly connected may mean that the plurality of pressure application ports (310) are fluidly connected to the PCR reaction solution reservoir (122) and the oil reservoir (111). As described above, the plurality of pressure application ports (310) are fluidly connected to the PCR reaction solution reservoir (122) and the oil reservoir (111) when the pump (200) generates positive pressure.

[0110]

[0111] [Example 1: Implementing the entire pressure applying pathway network as a rigid pipe]

[0112] Referring to FIGS. 3 to 5, in Example 1, the droplet generating device includes a pump (200) and a pressure applying structure. Here, the pressure applying structure includes a plurality of pressure applying ports (310) and a pressure applying path network (320) fluidly connecting the plurality of pressure applying ports (310) and the pump (200). In addition, although not shown in FIGS. 3 to 5, the droplet generating device may further include one or more motors. The droplet generating device may control one or more motors connected to the pressure applying structure to move the plurality of pressure applying ports (310) and / or the droplet generating cartridge (100).

[0113] In the following description, the meaning that the plurality of pressure application ports (310) and / or the droplet generation cartridge (100) move may mean that the droplet generation device controls one or more motors to move the plurality of pressure application ports (310) and / or the droplet generation cartridge (100). In addition, the meaning that the pump (200) generates pressure may mean that the droplet generation device controls the pump (200) to generate pressure.

[0114]

[0115] In Example 1, as shown in FIG. 3, a pressure application path network (320) fluidly connecting a plurality of pressure application ports (310) and a pump (200) is implemented entirely using rigid pipes. In this case, the pressure application path network (320) may be implemented using a single rigid pipe or may be implemented using two or more rigid pipes. For example, a portion of the pressure application path network (320) may be implemented using a first rigid pipe, and the remaining portion may be implemented using a second rigid pipe.

[0116] In Example 1, as shown in FIG. 4, the pressure application path network (320) is implemented entirely as a rigid pipe, and a part of the pressure application path network (320) is implemented as a length-fixed rigid pipe, and the remaining part is implemented as a length-variable rigid pipe. In addition, in Example 1, as shown in FIG. 5, the pressure application path network (320) is implemented entirely as a length-variable rigid pipe.

[0117] Fig. 4(a) shows one implementation example according to Embodiment 1. Referring to Fig. 4(a), in Embodiment 1, the pressure application path network (320) includes an air conduit network (321), an air conduit (322), and a valve (323).

[0118] The air pipe network (321) fluidly connects between the plurality of pressure application ports (310) and the valve (323), and the air pipe (322) fluidly connects between the valve (323) and the pump (200). The valve (323) is for opening and closing the pressure application path network (320). When the droplet generation device opens the valve (323), the pressure generated by the pump (200) is transmitted to the plurality of pressure application ports (310) through the pressure application path network (320). On the other hand, when the droplet generation device (323) closes the valve, the pressure generated by the pump (200) is not transmitted to the plurality of pressure application ports (310). That is, the valve (323) can allow the pressure generated from the pump (200) to be applied to the droplet generation cartridge (100) through the plurality of pressure application ports (310).

[0119] Meanwhile, in Example 1, the pressure generated by the pump (200) is transmitted to the plurality of pressure application ports (310), and the pressure generated by the pump (200) is applied to the droplet generation cartridge (100), which implies that the droplet generation device has opened the valve (323).

[0120]

[0121] In the implementation example of Example 1 according to FIG. 4(a), the air pipe network (321) may be implemented as a fixed-length rigid pipe, and the air pipe (322) may be implemented as a variable-length rigid pipe. In addition, the length of the first portion (322a) of the air pipe (322) may be variable along the y-axis, and the length of the second portion (322b) of the air pipe (322) may be variable along the x-axis.

[0122] Referring to Fig. 4(a), the droplet generating cartridge (100) can move inside the droplet generating device along the direction (1). In addition, the plurality of pressure application ports (310) can move to a first position on the upper portion of the droplet generating cartridge (100), and while the plurality of pressure application ports (310) move to the first position, the length of the second portion (322b) of the air tube (322) can increase along the direction (2).

[0123] Additionally, the length of the first portion (322a) of the air tube (322) may be lengthened in the direction (3) while the plurality of pressure application ports (310) move downward and move to a second position where the plurality of pressure application ports (310) are fluidly connected with the droplet generating cartridge (100).

[0124] As described above, the plurality of pressure application ports (310) can be moved to be fluidly connected with the droplet generation cartridge (100). In addition, the plurality of pressure application ports (310) can be moved in the opposite direction of the direction (3) to the first position and then moved in the opposite direction of the direction (2) to the third position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed.

[0125] While the plurality of pressure application ports (310) move in the opposite direction of direction (3) to the first position, the length of the first portion (322a) of the air tube (322) gradually becomes shorter. Also, while the plurality of pressure application ports (310) move in the opposite direction of direction (2) to the third position, the length of the second portion (322b) of the air tube (322) gradually becomes shorter.

[0126]

[0127] As another example, only the length of the first part (322a) of the air tube (322) implemented as a length-variable rigid pipe may be variable along the y-axis, and the length of the remaining part of the air tube (322) excluding the first part (322a) may be fixed.

[0128] In this case, the plurality of pressure application ports (310) may not move along the x-axis, but may only move along the y-axis. For example, in FIG. 4(a), when the droplet generation cartridge (100) moves into the droplet generation device along the direction (1), the plurality of pressure application ports (310) located at the first position move downward, and while the plurality of pressure application ports (310) move to the second position where they are fluidly connected to the droplet generation cartridge (100), the length of the first portion (322a) of the air tube (322) may increase along the direction (3).

[0129] Additionally, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed.

[0130] While the plurality of pressure application ports (310) move in the opposite direction of direction (3) to the first position, the length of the first portion (322a) of the air tube (322) gradually becomes shorter.

[0131]

[0132] In the implementation example of Example 1 according to FIG. 4(b), the air pipe network (321) may be implemented as a length-variable rigid pipe, and the air pipe (322) may be implemented as a length-fixed rigid pipe. In addition, the length of the first part (321a) of the air pipe network (321) may be variable along the y-axis, and the length of the second part (321b) of the air pipe network (321) may be variable along the x-axis.

[0133] Referring to Fig. 4(b), the droplet generating cartridge (100) can be moved inside the droplet generating device along the direction (1). In addition, a plurality of pressure application ports (310) can be moved to a first position on the upper portion of the droplet generating cartridge (100). While the plurality of pressure application ports (310) are moved to the first position, the length of the second portion (321b) of the air pipe network (321) can be lengthened along the direction (2).

[0134] Additionally, the length of the first portion (321a) of the air pipe network (321) can be lengthened in the direction (3) while moving downward until the plurality of pressure application ports (310) are positioned at a second position where they are fluidly connected to the droplet generating cartridge (100).

[0135] As described above, the plurality of pressure application ports (310) can move to be fluidly connected with the droplet generation cartridge (100). In addition, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position and then move in the opposite direction of the direction (2) to the third position to release the fluidic connection with the droplet generation cartridge (100) when droplet generation is completed. As the plurality of pressure application ports (310) move in the opposite direction of the direction (3) to the first position, the length of the first portion (321a) of the air pipe network (321) gradually becomes shorter. In addition, as the plurality of pressure application ports (310) move in the opposite direction of the direction (2) to the third position, the length of the second portion (321b) of the air pipe network (321) gradually becomes shorter.

[0136]

[0137] As another example, only the first part (321a) of the air tube (321) implemented as a length-variable rigid pipe may have a length that varies along the y-axis, and the remaining part of the air tube (321) excluding the first part (321a) may have a fixed length.

[0138] In this case, the plurality of pressure application ports (310) may not move along the x-axis, but may only move along the y-axis. For example, in FIG. 4(a), when the droplet generation cartridge (100) moves into the droplet generation device along the direction (1), the plurality of pressure application ports (310) located at the first position move downward, and while the plurality of pressure application ports (310) move to the second position where they are fluidly connected to the droplet generation cartridge (100), the length of the first portion (321a) of the air tube (321) may increase along the direction (3).

[0139] Additionally, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed.

[0140] While the plurality of pressure application ports (310) move in the opposite direction of direction (3) to the first position, the length of the first portion (321a) of the air tube (321) gradually becomes shorter.

[0141]

[0142] In the implementation example of Example 1 according to FIGS. 5(a) and (b), both the air pipe network (321) and the air pipe (322) can be implemented as length-variable rigid pipes. At this time, in FIG. 5(a), the length of a part (321b) of the air pipe network (321) can be varied on the x-axis, and the length of a part (322a) of the air pipe (322) can be varied on the y-axis. In FIG. 5(b), the length of a part (321a) of the air pipe network (321) can be varied on the y-axis, and the length of a part (322b) of the air pipe (322) can be varied on the x-axis.

[0143] Referring to FIGS. 5(a) and (b), the droplet generating cartridge (100) can move inside the droplet generating device along the direction (1). In addition, a plurality of pressure application ports (310) move to a first position on the top of the droplet generating cartridge (100), and while the plurality of pressure application ports (310) move to the first position on the top of the droplet generating cartridge (100), in FIG. 5(a), the length of a part (321b) of the air pipe network (321) can be lengthened along the direction (2), and in FIG. 5(b), the length of a part (322b) of the air pipe (322) can be lengthened along the direction (2).

[0144] Additionally, the plurality of pressure application ports (310) move downward until they are positioned at a second position where they are fluidly connected to the droplet generating cartridge (100), and while the plurality of pressure application ports (310) move to the second position, in FIG. 5(a) the length of the second portion (322a) of the air tube (322) may be lengthened along the direction (3), and in FIG. 5(b) the length of a portion (321a) of the air tube network (321) may be lengthened along the direction (3).

[0145] As described above, the plurality of pressure application ports (310) can move to be fluidly connected with the droplet generation cartridge (100). In addition, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position and then move in the opposite direction of the direction (2) to the third position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed. As the plurality of pressure application ports (310) move in the opposite direction of the direction (3) to the first position, the length of a portion (322a) of the air pipe (322) gradually becomes shorter in FIG. 5(a), and the length of a portion (321a) of the air pipe network (321) gradually becomes shorter in FIG. 5(b). In addition, as the plurality of pressure application ports (310) move in the opposite direction of direction (2) to the third position, the length of a part (321b) of the air pipe network (321) in FIG. 5(a) gradually becomes shorter, and the length of a part (322b) of the air pipe (322) in FIG. 5(b) gradually becomes shorter.

[0146]

[0147] Meanwhile, in Example 1, the length of the variable-length rigid pipe may be varied in units of a certain length according to the clock of the processor included in the droplet generating device and / or the unit angular velocity of one or more motors determined by the clock of the processor. That is, the length of the variable-length rigid pipe may not be varied to be shorter than a certain length unit.

[0148] Therefore, in order to implement Example 1, the first distance between the first position and the third position and / or the second distance between the first position and the second position may need to be determined according to the predetermined length unit.

[0149]

[0150] Meanwhile, in Example 1, the length of the variable-length rigid pipe changes as the plurality of pressure application ports (310) move. This change in length of the variable-length rigid pipe allows the fluid connection between the pump (200) and the plurality of pressure application ports (310) to be maintained by the change in length of the variable-length rigid pipe, regardless of the movement and position of the plurality of pressure application ports (310).

[0151]

[0152] [Example 2: Designing a portion of the pressure application path network as a rigid pipe]

[0153] Referring to FIGS. 6 and 7, in Example 2, the droplet generating device includes a pump (200) and a pressure applying structure. Here, the pressure applying structure includes a plurality of pressure applying ports (310) and a pressure applying path network (320) fluidly connecting the plurality of pressure applying ports (310) and the pump (200). In addition, although not shown in FIGS. 6 and 7, the droplet generating device may further include one or more motors. The droplet generating device may control one or more motors connected to the pressure applying structure to move the plurality of pressure applying ports (310) and / or the droplet generating cartridge (100).

[0154] In the following description, the meaning that the plurality of pressure application ports (310) and / or the droplet generation cartridge (100) move may mean that the droplet generation device controls one or more motors to move the plurality of pressure application ports (310) and / or the droplet generation cartridge (100). Additionally, the meaning that the pump (200) provides pressure may mean that the droplet generation device controls the pump (200) to provide pressure.

[0155]

[0156] In Example 2, as shown in FIG. 6, a part of a pressure application path network (320) fluidly connecting a plurality of pressure application ports (310) and a pump (200) is implemented as a rigid pipe, and the remaining part is implemented as a flexible tube.

[0157] Fig. 7 shows an implementation example according to Embodiment 2. Referring to Fig. 7, in Embodiment 2, the pressure application path network (320) includes an air conduit network (321), an air conduit (322), and a valve (323).

[0158] The air pipe network (321) fluidly connects between the plurality of pressure application ports (310) and the valve (323), and the air pipe (322) fluidly connects between the valve (323) and the pump (200). The description of the valve (323) is the same as the description of the valve in Example 1, and is therefore omitted.

[0159]

[0160] In the implementation example of Example 2 according to FIG. 7, the air pipe network (321) may be implemented as a rigid pipe, and the air pipe (322) may be implemented as a flexible tube.

[0161] Referring to FIG. 7, the droplet generating cartridge (100) can be moved inside the droplet generating device along the direction (1). In addition, the plurality of pressure applying ports (310) move to a first position on the top of the droplet generating cartridge (100). At this time, the air tube (322) (i.e., flexible tube) moves according to the movement of the plurality of pressure applying ports (310). For example, the air tube (322) can be deformed (e.g., the flexible tube gradually straightens) until the plurality of pressure applying ports (310) move along the direction (2) and are positioned at the first position on the top of the droplet generating cartridge (100).

[0162] Additionally, the plurality of pressure application ports (310) move downward in the direction (3) until they are positioned at a second position where they are fluidly connected to the droplet generating cartridge (100), and the air tube (322) (i.e., flexible tube) moves in accordance with the movement of the plurality of pressure application ports (310). In other words, the air tube (322) (i.e., flexible tube) is deformed in accordance with the movement of the plurality of pressure application ports (310).

[0163] As described above, the plurality of pressure application ports (310) can move to be fluidly connected with the droplet generation cartridge (100). In addition, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position and then in the opposite direction of the direction (2) to the third position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed. While the plurality of pressure application ports (310) move in the opposite direction of the direction (3) to the first position and then in the opposite direction of the direction (2) to the third position, the air tube (322) (i.e., the flexible tube) moves and deforms in accordance with the movement of the plurality of pressure application ports (310).

[0164]

[0165] As another example, referring to FIG. 7, when the droplet generating cartridge (100) moves inside the droplet generating device along the direction (1), the plurality of pressure application ports (310) located at the first position move downward along the direction (3) until they are located at the second position where they are fluidly connected to the droplet generating cartridge (100), and the air tube (322) (i.e., the flexible tube) moves along with the movement of the plurality of pressure application ports (310). In other words, the air tube (322) (i.e., the flexible tube) is deformed according to the movement of the plurality of pressure application ports (310).

[0166] As described above, when droplet generation is completed, the plurality of pressure application ports (310) can move in the opposite direction of the direction (3) to the first position to release the fluid connection with the droplet generation cartridge (100). While the plurality of pressure application ports (310) move in the opposite direction of the direction (3) to the first position, the air tube (322) (i.e., the flexible tube) moves and deforms according to the movement of the plurality of pressure application ports (310).

[0167]

[0168] Meanwhile, in Example 2, the flexible tube moves and deforms as the plurality of pressure application ports (310) move. This movement and deformation of the flexible tube ensures that the fluid connection between the pump (200) and the plurality of pressure application ports (310) is maintained regardless of the movement and position of the plurality of pressure application ports (310). In addition, the flexibility of the flexible tube helps the plurality of pressure application ports (310) move smoothly.

[0169]

[0170] [Example 3: Rigid Body]

[0171] Referring to FIGS. 8 and 9, in Example 3, the droplet generating device includes a pump (200) and a pressure applying structure (320), wherein a part of the pressure applying structure (320) is implemented as a rigid body (315) manufactured using a rigid material.

[0172] Additionally, although not shown in FIGS. 8 and 9, the droplet generating device may further include one or more motors. The droplet generating device may control one or more motors connected to the pressure applying structure to move the rigid body (315) and / or the droplet generating cartridge (100).

[0173] In the following description, the meaning that the rigid body (315) and / or the droplet generating cartridge (100) moves may mean that the droplet generating device controls one or more motors to move the rigid body (315) and / or the droplet generating cartridge (100). Additionally, the meaning that the pump (200) generates pressure may mean that the droplet generating device controls the pump (200) to generate pressure.

[0174]

[0175] Referring to FIGS. 8 (a) and (b), the rigid body (315) includes a plurality of pressure application ports (310), an airway network (325), and at least one pump connection port (322d) inside the rigid body (315). Referring to FIG. 8 (a), the airway network (325) includes an air pipe network (321), a valve (323), and a first air pipe (322c). The air pipe network (321) fluidly connects between the plurality of pressure application ports (310) and the valve (323), and the first air pipe (322c) fluidly connects between the valve and at least one pump connection port (324). The description of the valve (323) is omitted because it is the same as that described in Example 1.

[0176] Among the pressure application structure (300), the second air tube (322d), which is the remaining part excluding the rigid body, is implemented as a flexible tube, and the second air tube (322d) fluidly connects at least one pump connection port (324) and the pump (200).

[0177] Referring to FIG. 8(b), a plurality of pressure application ports (310) may be arranged on the lower surface of a rigid body (315) that can be fluidly connected to the droplet generating cartridge (100).

[0178]

[0179] FIG. 9 is for explaining that a plurality of pressure application ports (310) arranged on the lower surface of a rigid body (315) are fluidly connected to a droplet generating cartridge (100).

[0180] Referring to FIG. 9, the droplet generating cartridge (100) can move inside the droplet generating device along the direction (1). In addition, the rigid body (315) moves to a first position on the top of the droplet generating cartridge (100) so that the plurality of pressure application ports (310) are on the same axis as the droplet reservoir (113) (or the PCR reaction solution reservoir (112) and the oil reservoir (111)) of the droplet generating cartridge (100). At this time, the second air tube (322d) (i.e., the flexible tube) moves according to the movement of the rigid body (315). For example, the second air tube (322d) may be deformed (e.g., the flexible tube may be gradually straightened) until the rigid body (315) moves along the direction (2) and is positioned at the first position on the top of the droplet generating cartridge (100).

[0181] Additionally, the rigid body (315) moves downward in the direction (3) until it is positioned at a second position where it is fluidly connected to the droplet generating cartridge (100), and the second air tube (322d) (i.e., flexible tube) moves according to the movement of the rigid body (315). In other words, the second air tube (322d) (i.e., flexible tube) is deformed according to the movement of the rigid body (315).

[0182] At this time, the fact that the plurality of pressure application ports (310) are on the same axis as the droplet reservoir (113) (or the PCR reaction solution reservoir (112) and the oil reservoir (111)) of the droplet generation cartridge (100) may mean that the center of a specific pressure application port and the center of a droplet reservoir that must be fluidly connected to the specific pressure application port are located on the same axis.

[0183] As described above, the rigid body (315) can move so that the plurality of pressure application ports (310) are fluidly connected with the droplet generation cartridge (100). In addition, the rigid body (315) can move in the opposite direction of direction (3) to a first position and then in the opposite direction of direction (2) to a third position to release the fluid connection with the droplet generation cartridge (100) when droplet generation is completed. While the rigid body (315) moves in the opposite direction of direction (3) to the first position and in the opposite direction of direction (2) to the third position, the air tube (322) (i.e., the flexible tube) moves and deforms in accordance with the movement of the rigid body (315).

[0184]

[0185] As another example, referring to FIG. 8, when the droplet generating cartridge (100) moves inside the droplet generating device along the direction (1), the rigid body (315) located at the first position moves downward along the direction (3) until it is located at the second position fluidly connected to the droplet generating cartridge (100), and the air tube (322) (i.e., the flexible tube) moves along with the movement of the rigid body (315). In other words, the air tube (322) (i.e., the flexible tube) is deformed along with the movement of the rigid body (315).

[0186] When droplet generation is complete, the rigid body (315) can move in the opposite direction of direction (3) to the first position to release the fluid connection with the droplet generation cartridge (100). While the rigid body (315) moves in the opposite direction of direction (3) to the first position, the air tube (322) (i.e., the flexible tube) moves and deforms according to the movement of the rigid body (315).

[0187]

[0188] Meanwhile, as shown in Fig. 8, the exterior of the rigid body (315) may be a cubic shape. However, the exterior of the rigid body (315) is not limited to a cubic shape.

[0189] The rigid body (315) may include two sides that can be connected to a mechanical mechanism for movement of the rigid body (315), which is a component of the droplet generating device, and a lower surface that allows a plurality of pressure application ports (310) to be fluidly connected to the droplet generating cartridge (100).

[0190] In addition, among the multiple surfaces of the rigid body (315), at least one pump connection port (324) may be arranged on one surface (e.g., the rear surface, the side surface, the front surface, the upper surface, or the lower surface). For example, the pump connection port (324) may be arranged on the remaining surfaces except for the two sides to which the mechanical mechanism is connected and the lower surface on which the multiple pressure application ports (310) are formed, or may be arranged on the two side surfaces or the lower surface.

[0191]

[0192] Since the rigid body (315) has a three-dimensional shape including a plurality of surfaces, in order for the rigid body (315) to move smoothly in the direction (2) and / or the direction (3) of FIG. 9, it may be preferable that the second air pipe (322d) be implemented as a flexible tube. In addition, in order to maintain the fluid connection between the rigid body (315) and the pump (200) regardless of the position of the rigid body (315), it may be preferable that the second air pipe (322d) be a flexible tube. However, the present invention is not limited thereto, and as in Example 2, the second air pipe (322d) may also be implemented as a length-variable rigid pipe.

[0193] Meanwhile, the exterior of the rigid body (315) can be fabricated by processing a material having rigidity. In this case, the air pipe network (321) and the first air pipe (322c) inside the rigid body (315) can be created by drilling from one side of the rigid body (315) toward the opposite side. At this time, the air passage does not need to completely penetrate from one side to the opposite side. For example, the air pipe network (321) and the first air pipe (322c) can be formed in the form of a tunnel (or channel) inside the rigid body (315).

[0194] In this case, in order to fluidly connect the air pipe network (321) and the first air pipe (322c) and to place a component such as a valve (323) inside the rigid body (315), a part of the air pipe network (321) and / or a part of the first air pipe (322c) may be formed outside the rigid body (315). This will be described later.

[0195]

[0196] Meanwhile, when a rigid body (315) composed of a plurality of surfaces physically contacts a droplet generating cartridge (100) by moving along the direction (3) of FIG. 9, the plurality of pressure application ports (310) of the rigid body (315) and the droplet generating cartridge (100) can more stably maintain a fluidic coupling due to the pressing force of the rigid body (315). When the plurality of pressure application ports (310) and the droplet generating cartridge (100) stably maintain a fluidic coupling, the pressure applied to the plurality of pressure application ports (310) and the droplet generating cartridge (100) can be stable.

[0197] Accordingly, the material making up the rigid body (315) must have rigidity, and the mass of the rigid body (315) may be such that it can maintain a stable fluidic coupling with the droplet generating cartridge (100). For example, the material making up the rigid body (315) may have rigidity and a density that can maintain a stable fluidic coupling with the droplet generating cartridge (100) depending on the volume of the rigid body (315).

[0198] For example, the rigid body (315) can be created using a material having rigidity such as aluminum, alloy stell, steel, nickel alloys, ceramic, tungsten, copper, plastic (e.g., polyether ether ketone (PEEK), polyether ketone (PEK), polytetrafluoroethylene (PTFE)), or stainless steel.

[0199]

[0200] Additionally, the flexible tubes used in Examples 1 to 3 may be produced using materials having flexibility. For example, the flexible tubes may be produced using materials such as neoprene, silicone, nylon, polyurethane, PVC, or rubber.

[0201]

[0202] Meanwhile, the rigid pipe used in Examples 1 and 2 can be created using a material having rigidity, such as a rigid body (315).

[0203]

[0204] [Example 4: Droplet generating device for Examples 1 to 3]

[0205] FIGS. 10 to 11 are provided to explain examples of a droplet generating device (1000) to which any one of the above-described embodiments 1 to 3 can be applied.

[0206] Referring to FIGS. 10 and 11, the droplet generating device (1000) may include a pump (200), a pressure applying structure (300), a motor (400), a mechanical mechanism (500), a position sensor (600), a power source (700), and a controller (800).

[0207] The pressure applying structure (300) may be any one of the pressure applying structures of Embodiments 1 to 3 described above. Therefore, a detailed description of the pressure applying structure (300) will be omitted. Meanwhile, the valve (323) included in the pressure applying structure (300) may be a proportional valve or a solenoid valve. However, the present invention is not limited thereto, and any valve that operates to allow or not allow the pressure applied by the pump to be transmitted to a plurality of pressure applying ports (310) may be used as a valve included in the pressure applying structure (300) according to any one of Embodiments 1 to 3 of the present invention.

[0208] The motor (400) is mechanically connected to the mechanical mechanism (500) and can provide mechanical force to at least one mechanical component included in the mechanical mechanism (500) to move the pressure application structure (300) and / or the cartridge tray (510a). Here, the mechanical force provided by the motor (400) may be a rotational force. The rotational force provided by the motor (400) is converted into a linear force by the at least one mechanical component, and the converted linear force moves the pressure application structure (300) and / or the cartridge tray (510a) in a specific axial direction. At this time, the mechanical component that converts the rotational force of the motor (400) into a linear force is called a force conversion element.

[0209] For example, a linear force converted by the first force conversion element may move the pressure applying structure (300) in the y-axis direction (e.g., the up-and-down direction of the droplet generating device (1000)), and a linear force converted by the second force conversion element may move the cartridge tray (510a) in the x-axis direction (e.g., the front-back direction of the droplet generating device (1000)). At this time, the y-axis may be the up-and-down direction of the droplet generating device (1000), and the x-axis may be the front-and-back direction of the droplet generating device (1000). Accordingly, the y-axis may be an axis perpendicular to the x-axis. Meanwhile, the x-axis may be an axis parallel to the surface of the pressure applying structure (300) on which the plurality of pressure applying ports (310) are formed, and the y-axis may be an axis perpendicular to the surface of the pressure applying structure (300) on which the plurality of pressure applying ports (310) are formed.

[0210] For example, if a portion of the pressure applying structure (300) is formed of a rigid body (315) according to Example 3, the x-axis may be parallel to the lower surface of the rigid body (315) on which the plurality of pressure applying ports (310) are formed, and the y-axis may be perpendicular to the lower surface.

[0211] Additionally, the x-axis and y-axis of FIG. 11 may be the same as the x-axis and y-axis described in FIGS. 4, 5, 7, and 9. In other words, the direction (1) in Examples 1 to 3 may be one direction along the x-axis. For example, the direction (1) may be a direction from the outside to the inside of the droplet generating device (1000). Or, for example, the direction (1) may be a direction from the front to the rear of the droplet generating device (1000).

[0212] Additionally, the direction (2) in Examples 1 to 3 may be a direction opposite to one direction on the x-axis. For example, the direction (2) may be a direction from the inside to the outside of the droplet generating device (1000). Or, for example, the direction (2) may be a direction from the back to the front of the droplet generating device (1000).

[0213] Additionally, the direction (3) in Examples 1 to 3 may be one direction along the y-axis. For example, the direction (3) may be a direction from the top surface to the bottom surface of the droplet generating device (1000). Or, for example, the direction (3) may be a direction from the top cover of the droplet generating device (1000) to the bottom plate.

[0214] Meanwhile, the number of motors (400) according to the present disclosure may be one, but may also be two or more. For example, one motor may provide rotational force to the first force conversion element and the second force conversion element, or the first motor may provide rotational force to the first force conversion element and the second motor may provide rotational force to the second force conversion element.

[0215] The mechanical mechanism (500) may refer to a set of mechanical components for moving the pressure applying structure (300) along the y-axis and moving the droplet generating cartridge (100) along the x-axis using mechanical force provided from the motor (400).

[0216] The mechanical mechanism (500) may include a droplet generation movement mechanism (510) and a pressure application structure movement mechanism (520). The droplet generation movement mechanism (510) is a set of mechanical components for moving the droplet generation cartridge (100). For example, the droplet generation movement mechanism (510) may include a cartridge tray (510a), a cartridge tray rail (510b), and the second force conversion element (not shown).

[0217] The cartridge tray (510a) provides a plane on which the droplet generating cartridge (100) is placed. For example, the droplet generating cartridge (100) may be placed at a specific location on the upper surface of the cartridge tray (510).

[0218] The cartridge tray rail (510b) provides a path along which the cartridge tray (510a) can move. In other words, the cartridge tray rail (510b) helps the cartridge tray (510a) to move stably in the x-axis direction parallel to the rail path provided by the cartridge tray rail (510b).

[0219] The second force conversion element (not shown) converts the rotational force provided from the motor (400) into a linear force in the x-axis direction, as described above, to move the cartridge tray (510a). For example, the second force conversion element may be a lever, a screw, or a pulley.

[0220] The pressure application structure moving mechanism (530) is a set of mechanical components for moving the pressure application structure (300). For example, the pressure application structure moving mechanism (520) may include a side support (520a), a pressure application structure rail (520b), and the first force conversion element (not shown).

[0221] The side support (520a) includes an inner surface on which a pressure-applying structure rail (520b) is mounted. For example, the side support (520a) may include two supports, each of which is positioned opposite to the other. A pressure-applying structure rail (520b) is positioned on the inner surface of each support.

[0222] The pressure application structure rail (520b) is mechanically connected to both sides of the pressure application structure (300) to help the pressure application structure (300) move stably in a specific axial direction. For example, the pressure application structure rail (520b) included on each of the two inner sides of the side support (600) allows the pressure application structure (300) to move stably in the y-axis direction parallel to the rail path provided by the pressure application structure rail (520b). At this time, the pressure application structure rail (520b) may be mechanically connected to both sides of the pressure application structure (300).

[0223] For example, FIG. 12 illustrates the configurations of the rigid body (315) for connecting the rigid body (315) described in Example 3 with the pressure application structure rail (520b) and the motor (400). Referring to FIG. 12, the rigid body (315) may further include a rail coupling portion (316) for coupling the rigid body (315) with the pressure application structure rail (520b) for assisting the y-axis movement of the rigid body (315), and a motor coupling portion (317) for mechanically coupling the rigid body (315) with the motor (400). At this time, a first force conversion element may be mechanically coupled to the motor coupling portion (317). In other words, the motor coupling portion (317) may be mechanically coupled to the motor (400) through the first force conversion element.

[0224] Meanwhile, the pressure application structure (300) described in Examples 1 and 2 may further include a rail coupling portion (316) for connecting to the pressure application structure rail (520b) and a motor coupling portion (317) for mechanically coupling with the motor (400), like the rigid body (315) of Example 3, and may be mechanically coupled with the motor (400) and the pressure application structure rail (520b) according to the above-described description.

[0225] The first force conversion element (not shown) converts the rotational force provided from the motor (400) into a linear force in the y-axis direction, as described above, thereby moving the pressure application structure (300) in the y-axis direction. For example, the first force conversion element may be a lever, a screw, or a pulley.

[0226] Meanwhile, although not shown in FIGS. 10 and 11, the pressure application structure moving mechanism (530) may further include a side support rail and a third force conversion element. The side support rail may move the side support in the x-axis direction, thereby allowing the pressure application structure (300) to move in the x-axis direction. Meanwhile, the third force conversion element converts the rotational force provided from the motor (400) into a linear force in the x-axis direction, thereby providing a force capable of moving the side support (520a) in the x-axis direction. At this time, the third force conversion element may convert the rotational force provided from the first motor (400a) or the second motor (400b) into a linear force, or may convert the rotational force provided from a third motor different from the first motor (400a) and the second motor (400b) into a linear force.

[0227] The position sensor (600) can detect the position of the pressure application structure (300) on the y-axis.

[0228] The pump (200) generates pressure. In addition, the pump (200) is fluidly connected to a pressure application structure (300). The fluid connection between the pump (200) and the pressure application structure (300) allows pressure to be applied to the droplet generation cartridge (100) through a plurality of pressure application ports (310). In the present disclosure, the number of pumps (200) may be one or two or more. Meanwhile, the pump (200) according to an embodiment of the present disclosure may be a diaphragm pump. However, the present disclosure is not limited thereto, and any pump capable of providing negative or positive pressure to a plurality of pressure application ports (310) may be used as a pump according to an embodiment of the present disclosure.

[0229] The power source (700) can provide power to enable the motor (400), pump (200), valve (323), position sensor (600), and controller (800) to operate. For example, the power source (700) can be electrically connected to each of the motor (400), pump (200), position sensor (700), and controller (800) to provide power.

[0230] Alternatively, for example, the power source (700) may be electrically connected to the controller (800) to provide power to the controller (800), and may provide power to the motor (400), valve (323), pump (200), and position sensor (700) electrically connected to the controller (800) through the controller (900).

[0231] The controller (800) can control the operation of the valve (323), the motor (400), the pump (200), and the position sensor (600). Referring to FIG. 10, the controller (800) can be electrically connected to the pump (200), the motor (400), the valve (323), the position sensor (600), and the power source (700).

[0232] The controller (800) receives power from the power source (700) and controls the operation of the droplet generating device (1000) described in the present disclosure. In order to control the operation of the droplet generating device (1000), the controller (800) can control the operation of the pump (200), the motor (400), the valve (323), and the position sensor (600).

[0233]

[0234] FIG. 13 shows an example in which the pressure applying structure (300) of Example 3 is mounted on the droplet generating device (1000) described through FIGS. 10 and 11. However, FIG. 12 shows an example in which Example 3 among Examples 1 to 3 described above is mounted on the droplet generating device (1000) for the sake of convenience of understanding, and the pressure applying structure (300) mounted on the droplet generating device (1000) is not limited to Example 3. That is, it goes without saying that the pressure applying structures (300) of Examples 1 to 3 can be mounted on the droplet generating device (1000) and moved according to the description of FIGS. 10 and 11 described above.

[0235] Referring to FIG. 13, as described above, the air pipe network (321) and the first air pipe (322c) inside the rigid body (315) can be created by drilling from one side of the rigid body (315) toward the opposite side, and in this case, in order to fluidically connect the air pipe network (321) and the first air pipe (322c) and to place a component such as a valve (323) inside the rigid body (315), a part of the air pipe network (321) and / or a part of the first air pipe (322c) is shown formed outside the rigid body (315). 322c of FIG. 13(a) is a part of the first air pipe (322c) formed outside the rigid body (315), and 321 of FIG. 13(b) is a part of the air pipe network (321) formed outside the rigid body (315).

[0236]

[0237] Figure 14 is for explaining the operation process of the droplet generating device (1000).

[0238] It should be understood that the operation process of the droplet generation device (1000) described below is performed by the user inputting a command to activate the power source (700) (e.g., power ON), so that the power source (700) supplies power to the controller (800), and the controller (800) controls the pump (200), the valve (323), the position sensor (600), and the motor (400). Therefore, it should be understood that the operation of the droplet generation device (1000) in the description described below is performed under the control of the controller (800).

[0239] Referring to FIG. 14, a motor (400) (e.g., a second motor) moves a cartridge tray (510a) to the outside of a droplet generating device (1000) along the x-axis direction of FIG. 11 (S1401). A user places a droplet generating cartridge (100) in a designated area within the cartridge tray (510a) (S1403). At this time, a fluid is stored in the fluid reservoir (110) of the droplet generating cartridge (100), a PCR reaction solution is stored in the PCR reaction solution reservoir (120) of the droplet generating cartridge (100), and the droplet reservoir (130) of the droplet generating cartridge (100) is empty.

[0240] The motor (400) (e.g., the second motor) moves the cartridge tray (510a) into the interior of the droplet generating device (1000) along the x-axis direction of FIG. 11 (S1405). For example, when the droplet generating device (1000) receives a command that the cartridge tray (510a) moves into the interior of the droplet generating device (1000), or when the droplet generating device (1000) detects that the droplet generating device (1000) is placed on the cartridge tray (510a), or when the droplet generating device (1000) (i.e., the controller (800)) counts a preset time after the cartridge tray (510a) moves to the outside of the droplet generating cartridge (100) and when the preset time elapses, the motor (400) (e.g., the second motor) moves the cartridge tray (510a) into the interior of the droplet generating device (1000).

[0241] A motor (400) (e.g., a first motor) moves a plurality of pressure application ports (310) or a rigid body (315) to move the plurality of pressure application ports (310) to the upper surface of the droplet generation cartridge (100) (S1407). Accordingly, the plurality of pressure application ports (310) are fluidly connected to the droplet reservoir (113) (when negative pressure is provided) or the PCR reaction solution reservoir (112) and the oil reservoir (111) (when positive pressure is provided). Meanwhile, the movement process and operation for moving the plurality of pressure application ports (310) to the upper surface of the droplet generation cartridge (100) and fluidly connecting them to the droplet generation cartridge (100) have already been described in Embodiments 1 to 3, and thus, a detailed description thereof will be omitted.

[0242] When the droplet generating device (1000) detects through the position sensor (600) that the plurality of pressure application ports (310) or the rigid body (315) have moved to the second position on the y-axis, the motor (400) (e.g., the first motor) stops operating so that the plurality of pressure application ports (310) or the rigid body (315) are fixed to the second position. For example, when the controller (900) receives position information of the plurality of pressure application ports (310) or the rigid body (315) from the position sensor (600), and the received position information indicates that the plurality of pressure application ports (310) or the rigid body (315) have moved to the second position on the y-axis, the controller (900) stops operating the motor (400) (e.g., the first motor) so that the plurality of pressure application ports (310) or the rigid body (315) are fixed to the second position.

[0243] Here, the second position is the same as the second position described in Examples 1 to 3, and is a position that allows the plurality of pressure application ports (310) and the droplet generating cartridge (100) to be fluidly coupled.

[0244] The pump (200) generates pressure (negative or positive pressure). In addition, when the droplet generation device (1000) opens the valve (323), the pressure generated by the pump (200) is applied to the droplet generation cartridge (100) through a plurality of pressure application ports (310) (S1409).

[0245] For example, the pump (200) can apply negative pressure to the droplet reservoir (130) through a plurality of pressure application ports (310), or can apply positive pressure to the oil reservoir (110) and the PCR reaction liquid reservoir (120) through a plurality of pressure application ports (310).

[0246] Here, the specific operation process in which the pump (200) generates pressure and the droplet generating device (1000) opens the valve (323) will be described later.

[0247] When the pressure generated from the pump (200) is applied to the droplet generating cartridge (1000), as described in FIG. 1, the fluid and the PCR reaction liquid meet to generate droplets, and the generated droplets are stored in the droplet storage (113) (S1411).

[0248]

[0249] [Example 5: Method of applying pressure to a droplet generating cartridge (100)]

[0250] In Examples 1 to 3, a pressure applying structure (300) capable of stably applying pressure to a droplet generating cartridge (100) is proposed.

[0251] In addition, in the description of S1409 of the above-described FIG. 14, it was explained that the pump (200) generates pressure, and when the valve (323) opens, the pressure generated by the pump (200) is applied to the droplet generating cartridge (100) through the plurality of pressure application ports (310).

[0252] Meanwhile, in order for the droplet generation device (1000) to stably apply pressure to the droplet generation cartridge (100), an operation of controlling the pump (200) and valve (323) to stably apply pressure may be further required.

[0253] Therefore, in Example 5, a method for controlling the pump (200) and the valve (323) of the droplet generating device (1000) to stably apply pressure to the droplet generating cartridge (100) will be examined. As described above, the control of the pump (200) and the valve (323) can be performed by the controller (800) of the droplet generating device (1000).

[0254] Fig. 15 is intended to explain a control method of a pump (200) and a valve (323) for stably applying pressure. However, as described above, it is not limited thereto, and even when the pump (200) generates positive pressure, the operation control of the pump (200) and the valve (323) can be performed in the same manner.

[0255] As previously described in FIG. 14, when the plurality of pressure application ports (310) or the rigid body (315) move to the upper surface of the droplet generating cartridge (100) (see S1407), the valve (330) is closed, the pump (200) is turned OFF, and no pressure is generated (FIG. 15(a)).

[0256] When the plurality of pressure application ports (310) or the rigid body (315) are moved to the upper surface of the droplet generation cartridge (100), and the droplet generation cartridge (100) and the plurality of pressure application ports (310) are fluidly coupled, the pump (200) is turned on at time t0 to generate pressure. However, even at this time, the valve (330) is still closed (Fig. 15(b)). At this time, t0 may be a time point after a first time has elapsed from the time when the plurality of pressure application ports (310) are fluidly coupled with the droplet generation cartridge (100). For example, the first time may be 0 seconds or may be several seconds greater than 0.

[0257] At t1, when a second time has elapsed from t0, the valve (323) opens, and the pressure generated by the pump (200) can be applied to the droplet generating cartridge (100) through a plurality of pressure application ports (310) (Fig. 15(c)). Here, for example, the second time can be several seconds and can be a preset time.

[0258] The valve (323) may be closed at t2, a third time period after t1 (Fig. 15(d)), and may be opened again at t3, a fourth time period after t2 (Fig. 15(e)). Furthermore, the valve (323) may be closed again at t4, a fifth time period after t3, and the pump (200) may stop operating to stop pressure generation (Fig. 15(f)). Here, for example, each of the third to fifth times periods may be several seconds and may be a preset time period.

[0259] However, the process of Fig. 15 (d) and Fig. 15 (e) may be optional. That is, after proceeding to Fig. 15 (c), Figs. 15 (d) to 15 (e) may be omitted, and at t2, the valve (323) may be closed and the pump (200) may stop operating to stop pressure production.

[0260] After t1, the valve (323) is open, and the pump (200) can still generate a constant pressure. If the pump (200) generates a negative pressure, the air pipe network (321) between the plurality of pressure application ports (310) and the valve (323) can maintain a first pressure (e.g., atmospheric pressure) from t0 to t1. In addition, the second pressure of the air pipe (322) between the valve (323) and the pump (200) can decrease from t0 to t1. For example, the second pressure of the air pipe (322) can approach a vacuum from t0 to t1. When the valve (323) is opened at t1, the pressure of the air pipe network (321) and the pressure of the air pipe (322) reach equilibrium in the time interval between t1 and t2. At this time, the time required for the pressure of the air pipe network (321) and the pressure of the air pipe (322) to reach equilibrium at t1 when the valve (323) is opened may be a very short time of less than 2 seconds.

[0261] Meanwhile, by closing the valve (323) again at t2 and the pump (200) continuing to generate pressure, the pressure in the air pipe (322) may decrease again from t2 to t3. For example, the pressure in the air pipe (322) may become closer to a vacuum from t2 to t3. When the valve (323) is opened again at t3, the pressure in the air pipe network (321) and the pressure in the air pipe (322) reach equilibrium again. At this time, the pressure in the air pipe network (321) may have become lower than the first atmospheric pressure due to the negative pressure transmitted between t1 and t2, and thus may reach equilibrium in a shorter time than the time taken between t1 and t2.

[0262] On the other hand, if the pump (200) generates positive pressure, the air pipe network (321) between the plurality of pressure application ports (310) and the valve (323) can maintain a first pressure (e.g., atmospheric pressure) from t0 to t1. In addition, the second pressure of the air pipe (322) between the valve (323) and the pump (200) can increase from t0 to t1. For example, the second pressure of the air pipe (322) can become greater than the first pressure from t0 to t1. When the valve (323) is opened at t1, the pressure of the air pipe network (321) and the pressure of the air pipe (322) reach equilibrium in the time interval between t1 and t2. At this time, the time required for the pressure of the air pipe network (321) and the pressure of the air pipe (322) to reach equilibrium at t1 when the valve (323) is opened may be a very short time of less than 2 seconds.

[0263] Meanwhile, by closing the valve (323) again at t2 and allowing the pump (200) to continue generating pressure, the pressure in the air pipe (322) can increase again from t2 to t3. In addition, when the valve (323) is opened again at t3, the pressure in the air pipe network (321) and the pressure in the air pipe (322) reach equilibrium again. At this time, the pressure in the air pipe network (321) will have become higher than the first atmospheric pressure due to the negative pressure transmitted between t1 and t2, and thus can reach equilibrium in a shorter time than the time taken between t1 and t2.

[0264] Example 5 prevents vibrations and large pressure fluctuations that occur when the pump (200) starts to generate pressure (i.e., when the pump (200) is driven) from being transmitted to the droplet generating cartridge (100).

[0265] When the pump (200) begins to generate pressure, a significant amount of vibration occurs in the pump (200), and thus a relatively large and unstable pressure may be applied to the droplet generating cartridge (100). However, after a certain period of time has passed since the pump (200) began to generate pressure, the operation of the pump (200) becomes stable, the vibration decreases, and a stable pressure may be applied.

[0266] Therefore, if the valve (323) is opened before the operation of the pump (200) becomes stable, pressure may not be stably applied to the droplet generation cartridge (100) before and after the operation of the pump (200) becomes stable, and thus the sizes of the generated droplets may not be uniform. In addition, due to the large pressure generated when the pump (200) begins to generate pressure, the PCR reaction solution and oil may move too quickly to the droplet reservoir (113), and thus the droplets may not be properly generated.

[0267] Therefore, to prevent vibration and unstable high pressure from being provided to the droplet generation cartridge (300) when the pump (200) begins to apply pressure, the droplet generation device (1000) keeps the valve (323) closed until t1, and then opens the valve (323) after t1 (i.e., after the operation of the pump (200) has stabilized). This prevents droplets from being generated before the pump (200) has stabilized, thereby ensuring uniformity in size between droplets.

[0268]

[0269] Hereinafter, examples of experiments are described in which the inventors of the present application used pressure applying structures (300) designed by the inventors to determine whether the size of droplets is uniformly generated when using each pressure applying structure (300).

[0270]

[0271] [Design Example 1]

[0272] The present disclosure describes design example 1 with reference to FIG. 16.

[0273] The pump (200) is a diaphragm pump. The negative pressure generated by the diaphragm pump is 0.56 bar.

[0274] Additionally, a portion of the air pipe network (321a) is a 10 mm stainless steel (SUS304) pipe. Additionally, the air pipe (322) is a 15 mm stainless steel (SUS304) pipe.

[0275] The valve (323) is a proportional valve. Eight pressure application ports (310) are formed by drilling eight holes in the lower surface of a plastic plate, and the remaining portion (321b) of the air pipe network is formed within the plate. A portion (321a) of the air pipe network and the remaining portion (321b) are connected via a connection port.

[0276]

[0277] [Design Example 2]

[0278] The present disclosure describes design example 2 with reference to FIG. 16.

[0279] It is the same as design example 1 except that a part of the air pipe network (321a) is a 15 mm stainless steel (SUS304) pipe.

[0280]

[0281] [Design Example 3]

[0282] The present disclosure describes design example 3 with reference to FIG. 16.

[0283] It is the same as design example 1 except that a part of the air pipe network (321a) is a 20 mm stainless steel (SUS304) pipe.

[0284]

[0285] [Design Example 4]

[0286] The present disclosure describes design example 4 with reference to FIG. 16.

[0287] The design is the same as Example 1 except that the air tube (322a) is a 15 mm flexible urethane tube.

[0288]

[0289] [Design Example 5]

[0290] The present disclosure describes design example 5 with reference to FIG. 16.

[0291] It is the same as design example 1 except that a part of the air pipe network (321a) is a 15 mm stainless steel (SUS304) pipe and the air pipe (322a) is a 15 mm flexible urethane tube.

[0292]

[0293] [Design Example 6]

[0294] The present disclosure describes design example 6 with reference to FIG. 16.

[0295] It is the same as design example 1 except that a part of the air pipe network (321a) is a 20 mm stainless steel (SUS304) pipe and the air pipe (322a) is a 15 mm flexible urethane tube.

[0296]

[0297] [Design Example 7]

[0298] The present disclosure describes design example 7 with reference to FIG. 17.

[0299] The pump (200) is a syringe pump. The negative pressure generated by the syringe pump is XX Pa. The rigid body (315) is made of aluminum. The length of the air pipe network (321) formed inside the rigid body (315) between the pressure application ports and the valve is 61 mm, and the length of the first air pipe (322c) between the valve and the pump connection port is 30 mm.

[0300] The valve (323) is a PP valve (Pilot-Operated Proportional valve). The number of pressure application ports formed on the lower surface of the rigid body (315) is eight. The second air tube (322d) is a 21 mm flexible urethane tube.

[0301]

[0302] [Comparison Example 1]

[0303] The present disclosure describes Comparative Example 1 with reference to FIG. 16.

[0304] The design is the same as Example 1 except that a portion of the air pipe network (321a) is a 10 mm flexible urethane tube and the air pipe (322a) is a 15 mm flexible urethane tube.

[0305]

[0306] [Comparison Example 2]

[0307] The present disclosure describes Comparative Example 2 with reference to FIG. 16.

[0308] The design is the same as Example 1 except that a portion of the air pipe network (321a) is a 15 mm flexible urethane tube and the air pipe (322a) is a 15 mm flexible urethane tube.

[0309]

[0310] [Comparison Example 3]

[0311] The present disclosure describes Comparative Example 3 with reference to FIG. 16.

[0312] The design is the same as Example 1 except that a portion of the air pipe network (321a) is a 20 mm flexible urethane tube and the air pipe (322a) is a 15 mm flexible urethane tube.

[0313]

[0314] [Experimental Method]

[0315] The experiment was conducted through the following process.

[0316] (1) A mixture of water (DI water) and PCR premix was dispensed into eight PCR reaction reservoirs of the droplet generation cartridge, and oil was dispensed into eight oil reservoirs (Well).

[0317] (2) Eight pressure application ports were physically brought into contact with eight droplet reservoirs (Wells) of the droplet generation cartridge.

[0318] (3) Voltage was supplied to the pump through the power supply to cause the pump to generate negative pressure.

[0319] (4) 10 seconds after supplying voltage to the pump, voltage was supplied to the valve through the power supply to open the valve.

[0320] (5) The valve was opened and, after 6 seconds, the voltage supply to the valve was cut off and the valve was closed again. At this time, the voltage supply to the pump was not cut off, and thus the negative pressure generation of the pump continued.

[0321] (6) 10 seconds after closing the valve, voltage was supplied to the valve again to open the valve.

[0322] (7) After 6 seconds of opening the valve again, the voltage supply to the pump and the voltage supply to the valve were cut off.

[0323] (8) The size of the droplets generated in eight oil reservoirs (Wells) was observed under a microscope, and images of the droplets were captured. At this time, the magnification of the images used in each experiment was the same.

[0324]

[0325] [Experimental Results]

[0326] [1] Experimental results of design example 1 (Fig. 18)

[0327] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1 1 13.76 1 13.54 1 19.35 1 18.5 1 1 11.67 1 15.86 1 15.45 3.02 2.61 Well 2 1 0 9.01 1 0 6.60 1 0 7.13 1 0 3.92 1 0 5.53 1 0 7.16 1 0 6.56 1 7.21

[0328] [2] Experimental results of design example 2 (Fig. 19)

[0329] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1104.16107.21105.28106.41109.27106.16106.421.741.64Well 2105.92108.14104.11105.11109.21107.75106.711.971.84

[0330] [3] Experimental results of design example 3 (Fig. 20)

[0331] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1108.76 114.37 115.44 110.22 106.41 109.21 110.74 3.48 3.14 Well 2108.26 108.13 107.63 109.46 110.45 108.01 108.66 1.07 0.99

[0332] [4] Experimental results of design example 4 (Fig. 21)

[0333] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1109.74109.64112.66111.40114.84107.41110.952.612.35Well 2108.72108.84109.90110.56106.02111.29109.221.851.70

[0334] [5] Experimental results of design example 5 (Fig. 22)

[0335] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1 1 17.0 1 1 14.26 1 1 1.85 1 10.57 1 12.08 1 12.66 1 13.07 2.27 2.01 Well 2 1 15.49 1 1 1.14 1 14.33 1 08.34 1 15.52 1 10.79 1 12.60 2.95 2.62

[0336] [6] Experimental results of design example 6 (Fig. 23)

[0337] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1 1 1 2.15 1 1 2.6 1 1 1 3.1 1 1 1 1.43 1 1 5.37 1 1 2.72 1 1 2.90 1.34 1.19 Well 2 1 1 2.70 1 1 4.73 1 1 8.88 1 1 4.23 1 1 0.58 1 1 8.07 1 1 4.87 3.16 2.75

[0338] [7] Experimental results of design example 7 (Figs. 24 to 27)

[0339] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1105.73 102.34 103.38 103.47 104.39 106.93 104.37 1.69 1.62 Well 2104.91 105.95 104.91 102.14 100.84 103.94 103.78 1.93 1.86 Well 3108.23 102.39 107.88 106.08 105.18 108.03 106.30 2.27 2.14 Well 4102.05105.89103.74103.47107.78106.61104.922.182.08Well 5107.71105.89106.06105.03105.62103.74105.681.301.23Well 6105.56107.69107.69107.88107.69105.89107.071.050.98Well 7100.4103.47103.38103.07105.01105.03103.391.691.64Well 8109.76108.65107.69105.54102.39105.62106.612.652.49

[0340] [8] Experimental results of comparative example 1 (Fig. 28)

[0341] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1 1 3 1.38 100.12 1 19.14 100.22 1 36.90 134.38 120.36 16.78 13.94 Well 2 1 0 3.40 107.30 118.48 132.14 126.19 131.38 119.82 12.28 10.25

[0342] [9] Experimental results of comparative example 2 (Fig. 29)

[0343] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1 1 2 4.9 2 1 0 1.3 2 1 3 7.8 4 1 0 1.8 0 9 6.9 4 1 3 1.4 0 1 5.7 0 17.7 4 1 5.3 3 Well 2 1 2 7.5 1 16 4.5 5 2 2 9.4 1 16 5 8 170.8 2 1 7 9.5 4 1 6 4.7 4 0.3 6 2 4.5 0

[0344]

[0010] Experimental results of comparative example 3 (Fig. 30)

[0345] Droplet 1 (um) Droplet 2 (um) Droplet 3 (um) Droplet 4 (um) Droplet 5 (um) Droplet 6 (um) Size Mean Standard Deviation CV (%) Well 1113.66110.87111.19119.16137.39115.99118.049.978.45Well 2135.52147.05146.45126.81219.84139.72152.5733.8022.16

[0346] [Analysis of Experimental Results]

[0347] [1] Analysis of experimental results of design examples 1 to 6 and comparative examples 1 to 3

[0348] Comparing the experimental results of Design Examples 1 to 3, it can be seen that the standard deviation of the droplet size does not decrease as the length of the stainless steel pipe between the eight pressure application ports and the valve decreases, and the standard deviation of the droplet size does not increase as the length of the stainless steel pipe increases. In other words, there appears to be no correlation between the length of the stainless steel pipe and the uniformity of the droplet size.

[0349] This is also true when comparing the experimental results of Design Examples 4 to 6. That is, the length of the stainless steel pipe does not affect the uniformity of the droplet size. The same holds true when comparing the experimental results of Comparative Examples 1 to 3. The length of the urethane tube does not affect the uniformity of the droplet size.

[0350] Accordingly, it can be seen that the length of the pipe or tube between the pressure application ports and the valve does not affect the droplet size.

[0351]

[0352] [2] Comparative analysis of the experimental results of design examples 1 to 3 and comparative examples 1 to 3.

[0353] Comparing the experimental results of Design Examples 1 to 3 and Comparative Examples 1 to 3, it can be seen that the standard deviation of the droplet sizes generated when the pressure application ports and the pump are connected through a stainless steel pipe is significantly smaller than the standard deviation of the droplet sizes generated when the pressure application ports and the pump are connected through a urethane tube. That is, the standard deviation of the droplet sizes in Comparative Examples 1 to 3 was approximately 10 to 40, and the standard deviation of the droplet sizes in Design Examples 1 to 3 was approximately 1 to 3.

[0354] This means that the standard deviation of the droplet size when the pressure application ports and the pump are stainless steel pipes is smaller in absolute value than the standard deviation of the droplet size when the pressure application ports and the pump are urethane tubes, but the range of standard deviation that can occur is also smaller.

[0355] This indicates that the size of the droplets generated is constant when the pressure application ports and the pump are stainless steel pipes, and the probability of the droplets being generated unevenly is also much lower.

[0356] Therefore, it was found that implementing a stainless steel pipe between the pressure application ports and the pump could produce a more stable and uniform droplet size than implementing a urethane tube.

[0357]

[0358] [3] Comparative analysis of the experimental results of design examples 3 to 6 and comparative examples 1 to 3.

[0359] Even when only some of the paths between the pressure application ports and the pump (i.e., the paths between the pressure application ports and the valve) were made of stainless steel pipes and the remaining paths were made of urethane tubes, the standard deviation of the sizes of the generated droplets was approximately 1 to 3. This is a much lower value than the standard deviation of approximately 10 to 40 of the sizes of the generated droplets when all paths between the pressure application ports and the pump were made of urethane tubes.

[0360] That is, it can be seen that even if only some of the paths between the pressure application ports and the pump are made of stainless steel pipes and the remaining paths are made of urethane tubes, a more uniform droplet size can be generated than if the entire path between the pressure application ports and the pump is made of urethane tubes.

[0361]

[0362] [4] Comparative analysis of experimental results of design examples 1 to 3 and design examples 4 to 6.

[0363] The standard deviation of the droplet sizes when the droplets were generated according to design examples 1 to 3 was approximately 1 to 3, and the standard deviation of the droplet sizes when the droplets were generated according to design examples 4 to 6 was also approximately 1 to 3. In other words, the standard deviation of the droplet sizes of design examples 1 to 3 and the standard deviation of the droplet sizes of design examples 4 to 6 are judged to be almost similar and not significantly different.

[0364] Accordingly, it was found that droplets can be generated sufficiently uniformly even if only some of the paths between the pressure application ports and the pump are made of stainless steel pipes and the remaining paths are made of urethane tubes, and it is not necessary to make all the paths of the paths of the pressure application ports and the pump of stainless steel pipes.

[0365] In particular, it can be seen that by configuring some of the paths between the pressure application ports and the pump, which are located closer to the pressure application ports than the pump (i.e., at least some of the paths between the pressure application ports and the valve), with stainless steel pipes, the droplet size can be generated uniformly.

[0366] Meanwhile, configuring some of the paths between the pressure application ports and the pump that are closer to the pump than the pressure application ports (i.e., at least some of the paths between the valve and the pump) with urethane tubes can be more helpful in facilitating the movement of the multiple pressure application ports.

[0367] Therefore, considering the mobility of multiple pressure application ports and the uniformity of the size of the generated droplets, it is considered desirable to configure some of the paths between the pressure application ports and the pump, which are closer to the pressure application ports than the pump, with rigid pipes, and to configure the remaining paths with flexible tubes.

[0368]

[0369] [5] Comparative analysis of the experimental results of Design Example 7 and Comparative Examples 1 to 3

[0370] The standard deviation of the droplet size when the droplets were generated according to Design Example 7 was approximately 1 to 2.5, and the standard deviation of the droplet size when the droplets were generated according to Comparative Examples 1 to 3 was approximately 10 to 40.

[0371] When droplets are generated using a rigid body of aluminum according to Design Example 7, it can be seen that the standard deviation of the droplet size is much smaller than when droplets are generated according to Comparative Examples 1 to 3.

[0372] That is, it can be seen that using a rigid body to generate droplets produces droplets with much more uniform size than constructing the path between the pressure application ports and the pump with urethane tubing.

[0373]

[0374] [6] Comparative analysis of the experimental results of Design Example 7 and Design Examples 1 to 6.

[0375] The standard deviation of the droplet size when the droplets were generated according to Design Example 7 was approximately 1 to 2.5, and the standard deviation of the droplet size when the droplets were generated according to Design Examples 1 to 6 was approximately 1 to 3.

[0376] In particular, it was found that the standard deviation of each Well in Design Example 7 was more uniform than the standard deviation of each Well in Design Examples 1 to 6.

[0377] That is, it can be seen that when a rigid body is used as in Design Example 7, the size of the generated droplets is more uniform than when at least part of the path between the pressure application ports and the pump is used as a rigid pipe.

[0378] This is believed to be because the mass of the rigid body is greater than the mass of the rigid pipe, and in Design Example 7, the pressure application ports are formed inside the rigid body, so the force with which the pressure application ports press the droplet generation cartridge is greater in Design Example 7 than in Design Examples 1 to 6.

[0379]

[0380] Although the embodiments described above have been described by way of limited examples and drawings, those skilled in the art will appreciate that various modifications and variations can be made based on the above teachings. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0381] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

In a droplet generating device that generates droplets using prepared PCR reaction solution and oil, A pump that provides negative pressure; A movable rigid body having a cubic shaped body and including a plurality of pressure applying ports and at least one pump connecting port, - Here, the plurality of pressure application ports and the at least one pump connection port are formed on the surface of the movable rigid body, The movable rigid body further includes an airway network formed inside the movable rigid body and connecting the plurality of pressure application ports and the at least one pump connection port; A flexible tube connecting the pump and at least one pump connection port; a motor that provides mechanical power; and A mechanical mechanism for moving the movable rigid body along the first axis using mechanical force provided from the motor; The flexibility of the flexible tube allows a fluid connection between the pump and the at least one pump connection port to be maintained regardless of the position of the movable rigid body, The rigidity of the above air passage network allows the negative pressure provided by the pump to be stably applied to the plurality of droplet generation reservoirs included in the droplet generation cartridge. Droplet generating device. In the first paragraph, The above droplet generating cartridge, It comprises a plurality of PCR reaction solution reservoirs into which PCR reaction solution is dispensed, a plurality of oil reservoirs into which oil is dispensed, and a plurality of droplet generation reservoirs, The negative pressure stably applied to the above multiple droplet generation reservoirs is: The PCR reaction solutions discharged from the plurality of PCR reaction solution reservoirs and the oils discharged from the plurality of oil reservoirs meet to create a liquid droplet, and the size of the droplets stored in the plurality of droplet generation reservoirs is kept constant. Droplet generating device. In the first paragraph, The plurality of pressure application ports are located on the lower surface of the movable rigid body so that negative pressure is applied to the plurality of droplet generation reservoirs. Droplet generating device. In the first paragraph, The above first axis is, Perpendicular to the surface on which the above multiple pressure application ports are formed, Droplet generating device. In the first paragraph, The above movable rigid body is, further comprising a valve that allows the negative pressure provided from the pump to be applied to the plurality of droplet generation reservoirs; The air passage network includes an air conduit network connecting the valve and the plurality of pressure application ports and an air conduit connecting the valve and the at least one pump connection port. Droplet generating device. In paragraph 5, The above droplet generating device, Further comprising a controller that controls the operation of the pump and the valve; The above controller, Control the pump so that the provision of the negative pressure begins at the first point in time, Opening the valve so that the negative pressure provided from the pump is applied to the plurality of droplet generation reservoirs after a first time interval from the first time point; Droplet generating device. In paragraph 6, The above first time interval is, preset, Droplet generating device. In the first paragraph, a position sensor for sensing the position of the movable rigid body; and Further comprising a controller for controlling the above motor; The above controller, Controlling the motor based on sensing of the position sensor so that the mechanical mechanism moves the movable rigid body downward along the first axis, such that the plurality of pressure application ports can be fluidly connected to the plurality of droplet generation reservoirs. Droplet generating device. In the first paragraph, a position sensor for sensing the position of the movable rigid body; and Further comprising a controller for controlling the above mechanical mechanism; The above controller, Controlling the motor based on sensing of the position sensor to cause the mechanical mechanism to move the movable rigid body upward along the first axis so that the fluid connection between the plurality of pressure application ports and the plurality of droplet generation reservoirs is released. Droplet generating device.

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