Droplet analysis device and method for cleaning droplet movement channel using droplet analysis device

Abstract

Disclosed is a droplet analysis device for analyzing droplets. Particularly, the droplet analysis device may comprise: a detection channel having a detection inlet and a detection outlet; a first valve connected to the detection outlet; a droplet suction unit; an X junction component; a hydrophobic fluid reservoir; a hydrophilic fluid reservoir; a first path adjustment unit for selectively connecting the droplet suction unit and the junction central inlet; a second path adjustment unit for selectively connecting the hydrophilic fluid reservoir and the at least two junction side inlets; a third path adjustment unit for selectively connecting the hydrophobic fluid reservoir and the at least two junction side inlets; and a hydraulic pressure supply unit for selectively supplying hydraulic pressure to the first to third path adjustment units.

Description

Droplet analyzer and method for the droplet analyzer to clean the droplet transport channel

[0001] The present disclosure relates to a droplet analysis device. More specifically, the present disclosure relates to a method for cleaning a channel through which a droplet travels within a droplet analysis device and a structure of a droplet analysis device for the same.

[0002] Polymerase Chain Reaction (PCR) is a molecular biological technique that replicates and amplifies desired portions of collected DNA. PCR allows researchers to selectively amplify specific DNA fragments from solutions containing extremely complex and trace amounts of DNA, such as the human genome. Additionally, it offers the advantages of a short amplification time, a simple experimental procedure, and the ability to perform amplification using fully automated machines. Consequently, PCR is widely used across various fields involving DNA processing, including molecular biology, medicine, forensic science, and biological classification.

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

[0004] Among these, digital PCR can analyze the status of the collected genes, the onset of disease, or infection by detecting fluorescence from each of the amplified droplets and counting the number of positive and negative droplets.

[0005] Meanwhile, among digital PCRs, there is droplet digital PCR (ddPCR), which detects the fluorescence of droplets contained in each of multiple wells by transferring them to a common droplet transfer channel. Since ddPCR transfers droplets contained in each of multiple wells to a single common droplet transfer channel, it causes a contamination problem in which droplets transferred from previous wells remain in the common droplet transfer channel. This contamination causes a problem where droplets remaining in the common droplet transfer channel are detected when detecting the fluorescence of droplets transferred from the next well, so a method to minimize this contamination problem is required.

[0006] The present disclosure relates to a droplet analysis device and a method for the droplet analysis device to clean a droplet movement channel.

[0007] The problem that the present disclosure aims to solve is to provide a droplet analysis device capable of solving the problem of contamination in a common droplet transport channel through which droplets contained in a plurality of wells all move.

[0008] In addition, this provides a droplet analysis device capable of ensuring the reliability of analysis results by resolving the problem of false positives and / or false negatives caused by droplets remaining in a common droplet transport channel.

[0009] The problems that the present disclosure aims to solve are not limited to those described above, and problems not mentioned will be clearly understood by those skilled in the art corresponding to the present disclosure from the present specification and the attached drawings.

[0010] A droplet analysis device for analyzing a droplet according to the present disclosure comprises: a detection channel having a detection inlet and a detection outlet; a first valve connected to the detection outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the detection channel; an X junction component having a junction center inlet, at least two junction side inlets and a junction outlet—wherein the junction outlet is connected to the detection inlet—; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first path control unit for selectively connecting the droplet suction device and the junction center inlet; a second path control unit for selectively connecting the hydrophilic fluid reservoir and the at least two junction side inlets; and a third path control unit for selectively connecting the hydrophobic fluid reservoir and the at least two junction side inlets. It may include a hydraulic supply unit that selectively supplies hydraulic pressure to the first to third path control units; and a controller that controls the operation of the first valve, the droplet suction unit, the first path control unit, the second path control unit, the third path control unit, and the hydraulic supply unit.

[0011] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0012] Additionally, the controller may close the first valve, control the second path control unit so that the hydrophilic fluid reservoir and the at least two junction side inlets are connected, control the first path control unit so that the droplet suction unit and the junction center inlet are connected, and control the hydraulic supply unit so that hydraulic pressure is supplied to the second path control unit, thereby performing the hydrophilic fluid cleaning operation.

[0013] Additionally, the controller may close the first valve, control the third path control unit so that the hydrophobic fluid reservoir and the at least two junction side inlets are connected, control the first path control unit so that the droplet suction unit and the junction center inlet are connected, and control the hydraulic supply unit so that hydraulic pressure is supplied to the third path control unit, thereby performing the first hydrophobic fluid cleaning operation.

[0014] A droplet analysis device for analyzing a droplet according to the present disclosure may include: a detection channel having a detection inlet and a detection outlet; a first valve connected to the detection outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the detection channel; an X junction component having a junction center inlet, at least two junction side inlets and a junction outlet—wherein the junction outlet is connected to the detection inlet—; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first pump providing hydraulic pressure to the junction center inlet; a second pump providing hydraulic pressure to the at least two junction side inlets; a second valve allowing the junction center inlet to be selectively connected to the droplet suction device or the first pump; and a controller for controlling the first valve, the second valve, the droplet suction device, the first pump, and the second pump.

[0015] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0016] Additionally, the controller may perform the hydrophilic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophilic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the junction center inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0017] Additionally, the controller may perform the first hydrophobic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the junction center inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0018] A droplet analysis device for analyzing a droplet according to the present disclosure comprises: a plate having a central inlet, at least two side inlets and an outlet—wherein the plate has an X junction, and the plate comprises: (i) a central channel extending between the central inlet and the X junction, (ii) at least two side channels extending between each of the at least two side inlets and the X junction, and (iii) a central outlet channel extending between the outlet and the X junction—; a first valve connected to the outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the plate; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first pump for providing hydraulic pressure to the central inlet; a second pump for providing hydraulic pressure to the at least two side inlets; and a second valve for allowing the central inlet to be optionally connected to the droplet suction device or the first pump. and a controller that controls the first valve, the second valve, the droplet suction device, the first pump, and the second pump; may be included.

[0019] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the plate and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the plate and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0020] Additionally, the controller may perform the hydrophilic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophilic fluid reservoir is connected to the second pump, controlling the second valve so that the droplet suction device is connected to the central inlet, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0021] Additionally, the controller may perform the hydrophobic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the central inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0022] According to embodiments of the present disclosure, the problem of contamination of a common droplet transport channel through which droplets contained in a plurality of wells all move can be solved.

[0023] Accordingly, the accuracy of counting positive and negative droplets can be improved, allowing for accurate analysis of the state of genes, the onset of disease, or infection.

[0024] The effects according to the present disclosure are not limited to the effects described above, and unmentioned effects will be clearly understood by those skilled in the art from the present disclosure and the attached drawings.

[0025] FIG. 1 is intended to explain the configuration of a droplet analysis device according to an embodiment of the present disclosure.

[0026] FIGS. 2 to 4 are intended to schematically illustrate the droplet transfer and washing operations in a droplet analysis device.

[0027] FIGS. 5 and 6 are intended to explain the contamination problem and the cause thereof resulting from the droplet transport and washing operations of a conventional droplet analysis device.

[0028] FIG. 7 is intended to explain the operation of a droplet analysis device according to an embodiment of the present disclosure.

[0029] FIGS. 8 to 17 are for explaining the operation of a droplet analysis device according to a first embodiment of the present disclosure.

[0030] FIGS. 18 to 27 are for explaining the operation of a droplet analysis device according to a second embodiment of the present disclosure.

[0031] FIGS. 28 to 41 are for explaining the operation of a droplet analysis device according to a third embodiment of the present disclosure.

[0032] FIGS. 42 to 54 are for explaining the operation of a droplet analysis device according to a fourth embodiment of the present disclosure.

[0033] FIGS. 55 to 67 are for explaining the operation of a droplet analysis device according to a fifth embodiment of the present disclosure.

[0034] FIGS. 68 to 80 are for explaining the operation of a droplet analysis device according to the sixth embodiment of the present disclosure.

[0035] FIGS. 81 to 84 are intended to explain the effect of cleaning a common droplet transport channel according to an embodiment of the present disclosure.

[0036] The aforementioned objects, features, and advantages of the present disclosure will become more apparent from the following detailed description in conjunction with the accompanying drawings. However, as the present disclosure is subject to various modifications and may have various embodiments, specific embodiments are illustrated in the drawings and described in detail below.

[0037] The embodiments described in this specification are intended to clearly explain the concept of this disclosure to those skilled in the art to which this disclosure belongs. Therefore, this disclosure is not limited by the embodiments described in this specification, and the scope of this disclosure should be interpreted to include modifications or variations that do not deviate from the concept of this disclosure.

[0038] The drawings attached to this specification are intended to facilitate the explanation of the present disclosure, and the shapes depicted in the drawings may be exaggerated as necessary to aid in understanding the present disclosure; therefore, the present disclosure is not limited by the drawings.

[0039] If it is determined that a detailed description of known functions or configurations related to the present disclosure could unnecessarily obscure the essence of the present disclosure, such detailed description is omitted. Furthermore, numbers used in the description of this specification (e.g., First, Second, etc.) are merely identifiers to distinguish one component from another.

[0040] Furthermore, the suffixes "unit," "module," and "part" used for components in the following description are assigned or used interchangeably solely for the ease of drafting the specification, and do not inherently possess distinct meanings or roles.

[0041]

[0042] A droplet analysis device for analyzing a droplet according to the present disclosure comprises: a detection channel having a detection inlet and a detection outlet; a first valve connected to the detection outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the detection channel; an X junction component having a junction center inlet, at least two junction side inlets and a junction outlet—wherein the junction outlet is connected to the detection inlet—; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first path control unit for selectively connecting the droplet suction device and the junction center inlet; a second path control unit for selectively connecting the hydrophilic fluid reservoir and the at least two junction side inlets; and a third path control unit for selectively connecting the hydrophobic fluid reservoir and the at least two junction side inlets. It may include a hydraulic supply unit that selectively supplies hydraulic pressure to the first to third path control units; and a controller that controls the operation of the first valve, the droplet suction unit, the first path control unit, the second path control unit, the third path control unit, and the hydraulic supply unit.

[0043] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0044] Additionally, the controller may close the first valve, control the second path control unit so that the hydrophilic fluid reservoir and the at least two junction side inlets are connected, control the first path control unit so that the droplet suction unit and the junction center inlet are connected, and control the hydraulic supply unit so that hydraulic pressure is supplied to the second path control unit, thereby performing the hydrophilic fluid cleaning operation.

[0045] Additionally, the controller may close the first valve, control the third path control unit so that the hydrophobic fluid reservoir and the at least two junction side inlets are connected, control the first path control unit so that the droplet suction unit and the junction center inlet are connected, and control the hydraulic supply unit so that hydraulic pressure is supplied to the third path control unit, thereby performing the first hydrophobic fluid cleaning operation.

[0046] Additionally, the controller performs a second hydrophobic fluid washing operation that allows the hydrophilic fluid filled between the X junction member and the second path control unit to move to the detection channel during the hydrophilic fluid washing operation; wherein the second hydrophobic fluid washing operation may be performed between the hydrophilic fluid washing operation and the first hydrophobic fluid washing operation.

[0047] In addition, the controller can perform the second hydrophobic fluid cleaning operation by opening the first valve, controlling the first path control unit so that the hydrophobic fluid reservoir and the junction center inlet are connected, and controlling the hydraulic supply unit to provide hydraulic pressure to the first path control unit.

[0048] Additionally, the controller may perform a first droplet transfer operation of transferring droplets contained in the designated well between the first path control unit and the X junction member; and a second droplet transfer operation of transferring droplets between the first path control unit and the X junction member to the detection channel.

[0049] Additionally, the controller may perform the first droplet transfer operation by moving the droplet suction device to the designated well, closing the first valve, controlling the third path control unit so that the at least two junction side inlets are connected, controlling the first path control unit so that the droplet suction device and the junction center inlet are connected, and controlling the hydraulic supply unit to provide hydraulic pressure to the third path control unit, and may perform the second droplet transfer operation by opening the first valve, controlling the first path control unit so that the droplet suction device and the junction center inlet are not connected, and controlling the hydraulic supply unit to provide hydraulic pressure to the first path control unit.

[0050] In addition, the controller may perform the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, and may also perform the first droplet transfer operation and the second droplet transfer operation after the hydrophobic fluid washing operation.

[0051] Additionally, prior to the hydrophilic fluid washing operation, the controller can move the droplet suction device to the first well during the first droplet transfer operation to transfer the droplets contained in the first well between the first path control unit and the X junction member.

[0052] Additionally, after the hydrophobic fluid washing operation, the controller can move the droplet suction device to a second well different from the first well during the first droplet transfer operation, thereby transferring the droplets contained in the second well between the first path control unit and the X junction member.

[0053] Additionally, the controller controls the droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, wherein the first position may be a position where a droplet storage unit including the designated well is not placed.

[0054] A droplet analysis device for analyzing a droplet according to the present disclosure may include: a detection channel having a detection inlet and a detection outlet; a first valve connected to the detection outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the detection channel; an X junction component having a junction center inlet, at least two junction side inlets and a junction outlet—wherein the junction outlet is connected to the detection inlet—; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first pump providing hydraulic pressure to the junction center inlet; a second pump providing hydraulic pressure to the at least two junction side inlets; a second valve allowing the junction center inlet to be selectively connected to the droplet suction device or the first pump; and a controller for controlling the first valve, the second valve, the droplet suction device, the first pump, and the second pump.

[0055] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0056] Additionally, the controller may perform the hydrophilic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophilic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the junction center inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0057] Additionally, the controller may perform the first hydrophobic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the junction center inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0058] Additionally, the controller performs a second hydrophobic fluid washing operation that allows the hydrophilic fluid filled between the X junction member and the second valve to move to the detection channel during the hydrophilic fluid washing operation; wherein the second hydrophobic fluid washing operation may be performed between the hydrophilic fluid washing operation and the first hydrophobic fluid washing operation.

[0059] Additionally, the controller can perform the second hydrophobic fluid cleaning operation by opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the junction center inlet and the first pump are connected, and controlling the first pump to provide hydraulic pressure to the junction center inlet.

[0060] Additionally, the controller may perform a first droplet transfer operation of transferring droplets contained in the designated well between the second valve and the X junction member; and a second droplet transfer operation of transferring droplets between the second valve and the X junction member to the detection channel.

[0061] Additionally, the controller can perform the first droplet transfer operation by moving the droplet suction device to the designated well, closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the junction center inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two junction side inlets.

[0062] Additionally, the controller can perform the second droplet transfer operation by opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the first pump and the junction center inlet are connected, and controlling the first pump so that the first pump provides hydraulic pressure to the junction center inlet.

[0063] In addition, the controller may perform the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, and may also perform the first droplet transfer operation and the second droplet transfer operation after the hydrophobic fluid washing operation.

[0064] Additionally, prior to the hydrophilic fluid washing operation, the controller may move the droplet suction device to the first well during the first droplet transfer operation to transfer the droplets contained in the first well between the second valve and the X junction member.

[0065] Additionally, after the hydrophobic fluid washing operation, the controller can move the droplet suction device in the first droplet transfer operation to a second well different from the first well, and transfer the droplets contained in the second well between the second valve and the X junction member.

[0066] Additionally, the controller controls the droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, wherein the first position may be a position where a droplet storage unit including the designated well is not placed.

[0067]

[0068] A droplet analysis device for analyzing a droplet according to the present disclosure comprises: a plate having a central inlet, at least two side inlets and an outlet—wherein the plate has an X junction, and the plate comprises: (i) a central channel extending between the central inlet and the X junction, (ii) at least two side channels extending between each of the at least two side inlets and the X junction, and (iii) an outlet channel extending between the outlet and the X junction—; a first valve connected to the outlet; a droplet suction device for drawing droplets contained in a designated well to transfer the droplets to the plate; a hydrophobic fluid reservoir for storing a hydrophobic fluid; a hydrophilic fluid reservoir for storing a hydrophilic fluid; a first pump for providing hydraulic pressure to the central inlet; a second pump for providing hydraulic pressure to the at least two side inlets; and a second valve for allowing the central inlet to be optionally connected to the droplet suction device or the first pump. and a controller that controls the first valve, the second valve, the droplet suction device, the first pump, and the second pump; may be included.

[0069] Here, the controller performs a hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the plate and the droplet suction device; and a first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the plate and the droplet suction device; wherein the hydrophilic fluid washing operation may be performed before the first hydrophobic fluid washing operation.

[0070] Additionally, the controller may perform the hydrophilic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophilic fluid reservoir is connected to the second pump, controlling the second valve so that the droplet suction device is connected to the central inlet, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0071] Additionally, the controller may perform the hydrophobic fluid cleaning operation by closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the central inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0072] Additionally, the controller performs a second hydrophobic fluid washing operation that allows the hydrophilic fluid filled between the X junction and the second valve to move to the plate during the hydrophilic fluid washing operation; wherein the second hydrophobic fluid washing operation may be performed between the hydrophilic fluid washing operation and the first hydrophobic fluid washing operation.

[0073] Additionally, the controller can perform the second hydrophobic fluid cleaning operation by opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the central inlet and the first pump are connected, and controlling the first pump to provide hydraulic pressure to the central inlet.

[0074] Additionally, the controller may perform a first droplet transfer operation of transferring droplets contained in the designated well between the second valve and the plate; and a second droplet transfer operation of transferring droplets between the second valve and the plate to the plate.

[0075] Additionally, the controller can perform the first droplet transfer operation by moving the droplet suction device to the designated well, closing the first valve, controlling the second pump so that the hydrophobic fluid reservoir and the second pump are connected, controlling the second valve so that the droplet suction device and the central inlet are connected, and controlling the second pump so that the second pump provides hydraulic pressure to the at least two side inlets.

[0076] Additionally, the controller can perform the second droplet transfer operation by opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the first pump and the central inlet are connected, and controlling the first pump so that the first pump provides hydraulic pressure to the central inlet.

[0077] In addition, the controller may perform the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, and may also perform the first droplet transfer operation and the second droplet transfer operation after the hydrophobic fluid washing operation.

[0078] Additionally, prior to the hydrophilic fluid washing operation, the controller may move the droplet suction device to the first well during the first droplet transfer operation to transfer the droplets contained in the first well between the second valve and the plate.

[0079] Additionally, after the hydrophobic fluid washing operation, the controller can move the droplet suction device in the first droplet transfer operation to a second well different from the first well, thereby transferring the droplets contained in the second well between the second valve and the plate.

[0080] Additionally, the controller controls the droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, wherein the first position may be a position where a droplet storage unit including the designated well is not placed.

[0081]

[0082] [General Digital PCR]

[0083] Digital PCR exhibits a sensitivity approximately 1,000 times higher than real-time PCR and is suitable for analyzing target genes even in complex mixtures. The high sensitivity of digital PCR allows it to be more effective than the real-time PCR method in cases where the concentration of the target in the sample is very low.

[0084] In addition, digital PCR is highly advantageous for the quantification of target DNA compared to real-time PCR.

[0085] To perform digital PCR, processes are fundamentally required to divide an analyte sample (solution) containing DNA into predetermined volumes and to allow an amplification reaction of the target DNA to occur within the divided sample (solution). In particular, digital PCR methods can be classified in various ways depending on the method of dividing the sample into predetermined volumes. The present disclosure relates to droplet digital PCR, which uses a method of dividing the sample into microdroplets.

[0086]

[0087] To perform droplet digital PCR, the following three processes are largely followed.

[0088]

[0089] (1) Droplet generation step

[0090] (a) a sample containing target DNA to be amplified and (b) a PCR reaction solution comprising i) primers for amplifying the target DNA, ii) a fluorescent probe or fluorescent dye, iii) dNTP (deoxynucleoside triphosphate) used as a material for the amplicon, iv) polymerase, etc. are prepared. For example, fluorescent probes or fluorescent dyes may include FAM, HEX, VIC, TAMRA, EvaGreen, etc. Additionally, the sample refers to the result of pretreatment necessary for PCR analysis of blood, tissue, cells, saliva, etc. collected from a subject. In particular, the sample contains DNA to be analyzed.

[0091] PCR reaction solution and oil are injected into the droplet generation cartridge. For example, the droplet generation cartridge may be divided into a first space where the PCR reaction solution is injected, a second space where the oil is injected, and a third space where the generated droplets are stored.

[0092] When pulling pressure is applied to the third space of the droplet generation cartridge, the PCR reaction solution injected into the first space and the oil injected into the second space move into the third space, generating droplets. The method of generating droplets using the PCR reaction solution and oil is based on microfluidics technology, but is not limited to this.

[0093]

[0094] (2) Droplet amplification step

[0095] As described above, after the droplets are generated, repetitive heat treatment is performed on the generated droplets according to a predetermined thermal cycle. Accordingly, the target DNA can be amplified within the PCR reaction solution contained in each droplet. Of course, DNA amplification occurs within droplets containing the target DNA, but not within droplets that do not contain the target DNA. Basically, the aforementioned positive droplets refer to droplets containing the target DNA, and negative droplets refer to droplets that do not contain the target DNA.

[0096]

[0097] (3) Droplet detection step

[0098] Meanwhile, generally, when attempting to detect two or more target DNAs using two or more fluorescent materials, PCR reagents can be designed so that each of the different fluorescences corresponds to a different target DNA. That is, when the first target DNA is amplified, the fluorescence emitted from the amplified amplicon is formed in a first wavelength band, and when the second target DNA is amplified, the fluorescence emitted from the amplified amplicon is formed in a second wavelength band distinct from the first wavelength band. Thus, a PCR device having multiple fluorescence detectors can detect multiple target DNAs from a single sample.

[0099] In the case of digital PCR, since the PCR reaction takes place within a single droplet, detecting the type (wavelength) of fluorescence emitted from the droplet after the amplification reaction is complete provides information about the target DNA contained within that droplet. Therefore, for digital PCR equipment, it is important to accurately detect the type of fluorescence emitted from each specific droplet.

[0100] Fluorescence is detected from droplets (positive droplets) containing amplified target DNA 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 the fluorescence radiated as the output light interacts with the fluorescent material possessed by the droplets.

[0101] In this case, a device including a fluorescence detector can count droplets by fluorescent material according to the wavelength of fluorescence corresponding to the fluorescent material. 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 specific wavelengths. In other words, the detector may include a filter that passes only specific wavelengths, and accordingly, can detect only the wavelength emitted by a specific fluorescent material.

[0102] Accordingly, each of the multiple fluorescence detectors can detect distinct wavelengths, and the device can individually count the number of droplets having a fluorescent material corresponding to the wavelength detectable by each detector among the amplified droplets, based on the number of times each detector detects the wavelength corresponding to it.

[0103] As a specific example, if the intensity of fluorescence detected by the fluorescence detector exceeds a certain threshold, the counting number of droplets of fluorescent material corresponding to the wavelength detectable by the fluorescence detector can be increased.

[0104] The device can analyze the biological characteristics of a sample based on the number of droplets counted by fluorescent material.

[0105]

[0106] Meanwhile, the aforementioned droplet generation step, droplet amplification step, and droplet detection step may each be performed separately in different devices. For example, the droplet generation step may be performed by a first device, the droplet amplification step by a second device, and the droplet detection step by a third device.

[0107] As another example, the aforementioned droplet generation step, droplet amplification step, and droplet detection step may be performed integrally in a single specific device.

[0108] As another example, two of the aforementioned droplet generation step, droplet amplification step, and droplet detection step 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 a fifth device. As another example, the droplet generation step may be performed in a sixth device, and the droplet amplification step and the droplet detection step may be performed in a seventh device.

[0109]

[0110] Meanwhile, the droplet analysis device described in this disclosure is a device that performs a droplet detection step. However, such a droplet analysis device is not required to perform only the droplet detection step. That is, the droplet analysis device may be the third device described above, a specific device, a fifth device, or a seventh device. Therefore, the droplet analysis device is sufficient if it includes the configurations described in the present disclosure below and is capable of performing functions and operations. In other words, the droplet analysis device of this disclosure may include at least one other configuration other than the configurations described in this disclosure, and may also perform other functions and operations other than those described in this disclosure.

[0111]

[0112] [Droplet Analysis Device General]

[0113] FIG. 1 is a block diagram showing the configurations of a droplet analysis device (10) according to an embodiment of the present disclosure. In FIG. 1, the configurations connected by solid lines are electrically connected to each other. In addition, the configurations connected by dotted lines in FIG. 1 are fluidly connected to each other.

[0114] Referring to FIG. 1, the droplet analysis device (10) may include a controller (1000), a fluorescence detector (2000), a hydraulic supply unit (3000), a droplet suction device (4000), a path control unit (5000), a droplet detection channel (6000), a droplet processing unit (7000), a fluid storage unit (8000), and a droplet storage unit (9000).

[0115] Meanwhile, some components within the droplet analysis device (10) may be connected by channels. For example, a hydraulic supply unit (3000), a droplet suction unit (4000), a path control unit (5000), a droplet detection channel (6000), a droplet processing unit (7000), a fluid storage unit (8000), and a droplet storage unit (9000) are connected by channels, and fluid and / or droplets may move between some components through the channels. That is, the channels refer to paths through which droplets and / or fluids move within the droplet analysis device (10).

[0116] The controller (1000) can control the fluorescence detector (2000), the hydraulic supply unit (3000), the droplet suction unit (4000), and the path control unit (5000). For example, the controller (1000) can control the fluorescence detector (2000) to inject light into the droplet detection channel (6000) and to detect fluorescence emitted from the droplet. Additionally, the controller (1000) can control the hydraulic supply unit (3000) to provide positive pressure or negative pressure to at least one of the droplet detection channel (6000), the path control unit (5000), the droplet suction unit (4000), the fluid storage unit (8000), and the droplet storage unit (9000). Additionally, the controller (1000) can control the droplet suction device (4000) to move onto a designated well among the wells included in the droplet storage unit (9000) or onto a droplet processing unit (7000) or a channel fluidly connected to the droplet processing unit (7000). Additionally, the controller (1000) can control the path control unit (5000) to adjust the path of the droplet or fluid.

[0117] The fluorescence detection unit (2000) is for detecting fluorescence contained in droplets flowing in a droplet transport channel and may include a plurality of fluorescence detectors. At this time, the plurality of fluorescence detectors may be arranged along the longitudinal direction of the droplet detection channel (6000) (or the direction of movement of the droplets and / or fluid within the droplet detection channel). The fluorescence detector includes a light source for outputting light toward the droplet detection channel (6000) and a detector for detecting fluorescence emitted when the output light interacts with the fluorescent material of the droplet.

[0118] Multiple fluorescence detectors may be arranged in a line on one side of the droplet detection channel (6000) to detect the type of fluorescence emitted from droplets flowing through the droplet detection channel (6000). For example, when fluorescence is detected from droplets, each of the light sources (e.g., laser or LED) output through each of the fluorescence detectors (2000) has a different wavelength, and each of the fluorescence detectors (2000) detects the corresponding wavelength to determine the type of detected fluorescence.

[0119] The fluid storage unit (8000) is fluidically connected to the hydraulic supply unit (3000) and stores the fluid provided to the channel by the hydraulic supply unit (3000).

[0120] The hydraulic supply unit (3000) is for providing positive or negative pressure and may include at least one pump. The hydraulic supply unit (3000) may provide positive or negative pressure to at least one of the droplet detection channel (6000), path control unit (5000), droplet suction device (4000), fluid storage unit (8000), and droplet storage unit (9000).

[0121] Here, positive pressure refers to hydraulic pressure generated when the hydraulic supply unit (3000) pushes fluid into at least one of the droplet detection channel (6000), path control unit (5000), droplet suction device (4000), fluid storage unit (8000), and droplet storage unit (9000).

[0122] On the other hand, negative pressure refers to hydraulic pressure generated when the hydraulic supply unit (3000) draws fluid from at least one of the droplet detection channel (6000), path control unit (5000), droplet suction device (4000), fluid storage unit (8000), and droplet storage unit (9000).

[0123] In other words, the hydraulic pressure applied by the hydraulic supply unit (3000) to cause fluid to flow out from the hydraulic supply unit (3000) is positive pressure, and the hydraulic pressure applied by the hydraulic supply unit (3000) to cause fluid to flow into the hydraulic supply unit (3000) is negative pressure.

[0124] The operation of the hydraulic supply unit (3000) may be performed by a single pump (e.g., a first pump (3100)) or by two or more pumps (e.g., a first pump (3100) and a second pump (3200)). The operation of each hydraulic supply unit (3000) providing positive or negative pressure using one or more pumps will be specifically explained through the embodiments described below.

[0125] The droplet storage unit (9000) is a place for storing droplets and may include a well plate having a plurality of wells. The droplet storage unit (9000) may refer to the well plate itself, or to the plurality of wells or each of the plurality of wells. However, the droplet storage unit (9000) is not necessarily limited to the well plate or the plurality of wells having the well plate, as long as it can provide a space for storing already formed droplets. Droplets may be stored in at least some of the plurality of wells.

[0126] The droplet suction device (4000) is for drawing droplets stored in the droplet storage unit (9000) and may be a needle having a channel that provides a path for the droplets to move. The droplet suction device (4000) may move onto a designated well among a plurality of wells included in the droplet storage unit (9000). Alternatively, the droplet suction device (4000) may move to allow fluid and / or droplets to flow to the droplet processing unit (7000). Meanwhile, the droplet suction device (4000) may have an inner channel and an outer channel. The inner channel serves as a passage for drawing droplets stored in the droplet storage unit (9000) or for flowing fluid and / or droplets. The outer channel supports the inner channel and provides a passage for fluid to flow between the droplet suction device (4000) and the inner channel. The passage provided by the external channel allows fluid to be supplied to the tip portion of the internal channel through the external channel so that air is not introduced into the internal channel when a droplet is drawn into the internal channel.

[0127] The path control unit (5000) is intended to control the path through which fluid or droplets provided from the fluid storage unit (8000) or the droplet storage unit (9000) move. The path control unit (5000) may include at least one valve and / or at least one fitting. For example, the valve is a device that opens some of the ports among the plurality of ports of the valve and closes some of the ports so that fluid and / or droplets can flow through the open ports. At this time, the operation of the valve opening and closing the ports can be performed by the control of the controller (1000). That is, the valve is electrically connected to the controller (1000) and can be opened and closed by the control of the controller (1000).

[0128] Additionally, for example, the fitting may be a fitting with an X junction or a fitting with a T junction. The fitting may branch fluid and / or droplets entering through a port so that they exit through multiple ports, or allow fluid and / or droplets entering through multiple ports to merge and exit through a single port. In this case, such a fitting may be an X-shaped or T-shaped pipe and may not be electrically connected to the controller (1000). Therefore, the fitting may not be controlled by the controller (1000).

[0129] In other words, the path control unit (5000) electrically connected to the controller (1000) in FIG. 1 may be a valve, but the fitting included in the path control unit (5000) may not be electrically connected to the controller (1000).

[0130] In the description of the present disclosure, the term "port" may be replaced with the terms "inlet" and "outlet." An inlet refers to a port among a plurality of ports of a fitting or valve into which droplets and / or fluid flow when a hydraulic providing unit (3000) applies positive pressure to the port. Additionally, an outlet refers to a port among a plurality of ports of a fitting or valve into which droplets and / or fluid flow out when a hydraulic providing unit (3000) applies positive pressure to the port.

[0131] The droplet detection channel (6000) may be a tube or pipe that provides a path for droplets to flow. Alternatively, as described below, the droplet detection channel (6000) may be manufactured as a plate. Light from a fluorescence detection unit (2000) may be irradiated onto the droplet detection channel (6000). In other words, the droplet detection channel (6000) is part of the channel through which droplets move within the droplet analysis device (10), and the fluorescence detection unit (2000) can detect the fluorescence of the droplets passing through the droplet detection channel (6000). Here, the droplet detection channel (6000) may have a detection inlet through which droplets enter and a detection outlet through which droplets exit. That is, droplets may move from the detection inlet to the detection outlet and pass through the droplet detection channel (6000).

[0132] The droplet processing unit (7000) is a place where droplets or fluids that have passed through the droplet detection channel (6000) or the droplet suction device (4000) move. For example, the droplet processing unit (7000) is fluidically connected to the outlet of the droplet detection channel (6000) so that the detected droplets can be stored, recovered, or disposed of. Additionally, the droplet processing unit (7000) can store, recover, or dispose of the fluid discharged through the droplet suction device (4000) during the cleaning process.

[0133]

[0134] Meanwhile, in dPCR, droplets are stored in multiple wells of a droplet storage unit, and while the droplets are stored in each of the multiple wells, a fluorescence detector moves through each of the multiple wells to detect the fluorescence emitted from the droplets. Therefore, the droplets do not move from the multiple wells, and since the detection of droplets proceeds in different locations, the droplets stored in different wells do not mix with each other.

[0135] Therefore, droplets stored in a specific well are not detected during the detection process of droplets stored in other wells, and are relatively free from the problem of contamination of the droplet analysis device (10). However, in the case of dPCR, since fluorescence measurement is performed while the droplets are stored in the well, it is difficult to clearly separate and observe the fluorescence emitted by the droplets within the well, so the accuracy of the measurement may be reduced.

[0136] In contrast, in ddPCR, droplets stored in multiple wells of the droplet storage unit (9000) all move to the droplet detection channel (6000) through a common path. Accordingly, as the droplets move through the droplet detection channel (6000), the fluorescence emitted by the droplets is observed. Therefore, the fluorescence emitted by the droplets can be detected by relatively clearly separating them, so the accuracy of the measurement can be high. Hereinafter, the common path through which droplets move from the droplet aspirator (4000) to the droplet detection channel (6000) is referred to as the common movement channel of the droplets. For example, the common movement channel may be from the tip (e.g., the tip of a needle) of the droplet aspirator (4000) to the outlet of the droplet detection channel (6000). Here, the tip of the droplet aspirator (4000) is the tip of the internal channel of the droplet aspirator (4000).

[0137] That is, the common movement channel is a common channel through which all droplets stored in multiple wells of the droplet storage unit (9000) move from the droplet suction device (4000) to the outlet of the droplet detection channel (6000).

[0138] However, in ddPCR, since droplets stored in multiple wells of the droplet storage unit (9000) all pass through the common transport channel, some of the droplets from the previous well may remain due to reasons such as sticking to the inner wall of the common transport channel. That is, the common transport channel may be contaminated by droplets that did not pass through the common transport channel.

[0139] When droplets from the next well share a common transport channel, these remaining droplets detach from the inner wall and pass through the channel together with the droplets from the next well; consequently, the fluorescence from these remaining droplets may be measured together with that of the droplets in the next well. If target DNA is present in the remaining droplets, it may be detected as present in the droplets of the next well even if they are not, leading to a false positive. Conversely, if target DNA is not present in the remaining droplets, the amount of droplets containing target DNA may be judged to be relatively small due to the presence of the remaining droplets, resulting in a false negative where the target DNA level is judged to be below the threshold even though it is present in the droplets of the next well.

[0140] In other words, in ddPCR, since droplets from all wells all pass through a single common transport channel, there is a very high possibility of contamination problems occurring, and such contamination problems result in unreliable measurement accuracy of ddPCR.

[0141]

[0142] Therefore, a method is required to remove residual droplets in the common transport channel after fluorescence measurements have been performed on droplets in the previous well and before fluorescence measurements are performed on droplets in the next well.

[0143] Hereinafter, the present disclosure outlines the droplet transport operation in ddPCR and the conventional operation for removing residual droplets within the common transport channel.

[0144]

[0145] [Droplet transfer operation and cleaning operation of the droplet analysis device (10)]

[0146] FIGS. 2 to 4 are diagrams for schematically explaining the droplet transfer operation and the conventional cleaning operation of the droplet analysis device (10).

[0147] Referring to FIG. 2, a droplet analysis device (10), schematically illustrated to explain droplet transfer and cleaning operations, may include a fluorescence detection unit (2000), a hydraulic supply unit (3000), a droplet suction unit (4000), a path control unit (5000), a droplet detection channel (6000), a fluid processing unit (7000), a fluid storage unit (8000), and a droplet storage unit (9000). Meanwhile, although not illustrated in the drawing, the operation of the hydraulic supply unit (3000), the droplet suction unit (4000), and the path control unit (5000) as described above may be controlled by a controller (1000).

[0148] The path control unit (500) can selectively connect the hydraulic supply unit (3000) to the droplet suction unit (4000) or the droplet detection channel. Additionally, the droplet suction unit (4000) can be selectively positioned on the droplet storage unit (9000) or positioned to discharge fluid to the fluid processing unit (7000). That is, the droplet analysis device (10) is designed so that the droplet suction unit (4000) can move.

[0149] [1] Droplet transfer operation

[0150] FIG. 3 is a diagram illustrating an example of a droplet transfer operation. Referring to FIG. 3(a), the controller (1000) can control the path control unit (5000) to fluidly connect the hydraulic supply unit (3000) and the droplet suction unit (4000). Additionally, the controller (1000) can position the droplet suction unit (4000) in the first well of the droplet storage unit (9000) and control the hydraulic supply unit (3000) to provide negative pressure, thereby drawing the droplet stored in the first well to the path control unit (5000) and the hydraulic supply unit (3000) within the first channel between the droplet suction unit (4000) and the hydraulic supply unit (3000). At this time, the first channel between the droplet suction unit (4000) and the hydraulic supply unit (3000) is filled with oil, and as the hydraulic supply unit (3000) draws in the filled oil, the droplet stored in the first well can move together with the oil to the channel between the path control unit (5000) and the hydraulic supply unit (3000).

[0151] Referring to FIG. 3(b), the controller (1000) controls the path control unit (5000) to fluidly connect the hydraulic supply unit (3000) and the droplet detection channel (6000), and controls the hydraulic supply unit (3000) to provide positive pressure. The positive pressure provided by the hydraulic supply unit (3000) can push the droplet to pass through the second channel between the hydraulic supply unit (3000) and the droplet detection channel (6000). At this time, the second channel between the hydraulic supply unit (3000) and the droplet detection channel (6000) is filled with oil, and as the hydraulic supply unit (3000) pushes the oil filled in the second channel, the droplet can pass through the second channel together with the oil.

[0152] Meanwhile, when the droplet stored in the first well passes through the second channel between the hydraulic supply unit (3000) and the droplet detection channel (6000) according to the droplet transfer operation described above, the controller (1000) positions the droplet suction device (4000) at a second well different from the first well of the droplet storage unit (9000) and can repeat the droplet transfer operation described above. Accordingly, the droplets stored in the second well also move between the path control unit (5000) and the hydraulic supply unit (3000) within the first channel in the same way, and then pass through the second channel between the hydraulic supply unit (3000) and the droplet detection channel (6000). Therefore, the droplets of the first well and the droplets of the second well move through the first channel and the second channel in common. That is, the first channel and the second channel are common movement channels through which all the droplets stored in each well move.

[0153] Meanwhile, the number of wells included in the droplet storage unit (9000) may be very large. For example, the droplet storage unit (9000) may include a very large number of wells, such as 24, 96, 384, or 1536. Accordingly, the droplet transfer operation described above can be performed repeatedly as many times as the number of wells. That is, the droplet transfer operation can be performed repeatedly from tens to thousands of times.

[0154] [2] Common moving channel contamination problem

[0155] Meanwhile, in the droplet transfer operation described above, some of the droplets from the first well may not completely pass through the common transfer channel and remain. Additionally, the remaining portion of droplets from the first well may pass through together with the droplets from the second well when they pass through the common transfer channel and be measured. Furthermore, other portions of the remaining droplets from the first well may still remain within the common transfer channel.

[0156] However, the above-described droplet transfer operation is repeated from tens to thousands of times depending on the number of wells in the droplet storage unit (9000). Therefore, as the above-described droplet transfer operation is repeated, the degree of contamination of the common transfer channel may also increase.

[0157] If the degree of contamination in the common transport channel increases, the likelihood of false positives and / or false negatives being measured when measuring a droplet in a specific well may increase accordingly. That is, in the example described above, a measurement of the droplet in the first well may be incorrectly detected as a measurement result for the second well.

[0158]

[0159] To solve the above-mentioned problem, common transfer channels within the droplet analysis device (10) can be replaced between droplet transfers of each well or after transferring droplets of dozens or hundreds of wells. That is, a method of frequently replacing them using disposable common transfer channels can be considered.

[0160] This may be the most perfect method for removing residual droplets within the common moving channel. However, it is not realistic because the cost and time wasted in replacing the common moving channel are significant, and the difficulty of replacing the common moving channel within the droplet analysis device (10) is high.

[0161] Therefore, in order to solve the problem of contamination of the common transport channel in ddPCR, a "washing operation" to wash the common transport channel can be performed between the droplet transfer operation of the first well and the droplet transfer operation of the second well.

[0162]

[0163] [3] Conventional cleaning operation

[0164] Meanwhile, in order for droplets to pass through the common transfer channel, a hydrophobic fluid (e.g., oil), which is an immiscible fluid that does not mix with the droplets, was supplied to the common transfer channel. Accordingly, during the "cleaning" operation, the common transfer channel was cleaned by pushing out the remaining droplets in the common transfer channel with the hydrophobic fluid.

[0165] Figure 4 is intended to schematically illustrate this "washing operation".

[0166] Referring to FIG. 4(a), the controller (1000) can control the hydraulic supply unit (3000) to load a hydrophobic fluid (e.g., oil) from the fluid storage unit (8000), and the path control unit (5000) can control the hydraulic supply unit (3000) and the droplet suction unit (4000) to fluidly connect. Additionally, the controller (1000) can move the position of the droplet suction unit (4000) so that the fluid discharged from the droplet suction unit (4000) moves to the fluid processor (7000). The controller (1000) controls the hydraulic supply unit (3000) to provide positive pressure. By doing so, the loaded hydrophobic fluid moves through the first channel between the droplet suction unit (4000) and the hydraulic supply unit (300) by the positive pressure, and can push out the droplets remaining in the first channel by physical force. The remaining droplets pushed by the physical force of the hydrophobic fluid are moved to the fluid processing unit (7000).

[0167] Referring to FIG. 4(b), the controller (1000) can control the path control unit (5000) to fluidly connect the hydraulic supply unit (3000) and the droplet detection channel (6000). Additionally, the controller (1000) controls the hydraulic supply unit (3000) to provide positive pressure. As a result, the loaded hydrophobic fluid moves through the second channel between the hydraulic supply unit (3000) and the droplet detection channel (6000) by the positive pressure, and can push out the droplets remaining in the second channel by physical force. The remaining droplets pushed out by the physical force of the hydrophobic fluid move to the fluid processing unit (7000).

[0168]

[0169] [Discovery of problems with cleaning operations using hydrophobic fluids]

[0170] As described above, the applicant discovered a problem in which false positives and / or false negatives are still found even when a cleaning operation (hereinafter referred to as a hydrophobic fluid cleaning operation) is performed using a hydrophobic fluid.

[0171] Accordingly, to determine how thoroughly the common transport channel is cleaned when a hydrophobic fluid washing operation is performed, the applicant measured a high-concentration droplet, performed the aforementioned hydrophobic fluid washing operation, and conducted an NTC (No Template Control) measurement. At this time, the droplet was measured with a buffer capable of replacing actual DNA added.

[0172] Figure 5 shows the measurement results described above. For the measurement in Figure 5, high-concentration sample droplets and NTC droplets were prepared. Here, high-concentration sample droplets are droplets obtained by heat-treating and amplifying a droplet generated by mixing sample DNA, primers, and a premix. Additionally, NTC droplets are droplets obtained by heat-treating and amplifying a droplet generated by mixing primers and a premix without sample DNA.

[0173] Meanwhile, a single dot indicated in Fig. 5 represents the intensity of fluorescence measured from the sample. Since the NTC droplet has no target (i.e., DNA) to bind to, such as a primer, DNA amplification does not occur, and thus almost no fluorescence signal is generated, resulting in a very small fluorescence intensity value. In contrast, the sample droplet binds to DNA, such as a primer, and DNA amplification occurs, so a relatively strong fluorescence signal intensity value is generated.

[0174] Fig. 5(a) shows the results of measuring the fluorescence of sample droplets by moving high-concentration sample droplets to the droplet detection channel (6000) first. It also shows the fluorescence measurement results when measuring the fluorescence of NTC droplets after a washing operation using a hydrophobic fluid. As can be seen in the box in Fig. 5(a), noise, which consists of dots that cannot be measured when measuring NTC droplets, is observed. This noise can be seen as high-concentration sample droplets remaining unwashed within the common moving channel and then moving to the droplet detection channel (6000) to be measured when NTC droplets are measured.

[0175] Figure 5(b) shows the result of measuring the fluorescence of the NTC droplets of Figure 5(a), performing a washing operation using a hydrophobic fluid, measuring the fluorescence of high-concentration sample droplets once again, and measuring the fluorescence of the NTC droplets once again.

[0176] Referring to Fig. 5(b), when the fluorescence of the second NTC droplets is measured and the measurement results are examined, it can be seen that the noise has increased significantly compared to the measurement results of the fluorescence of the first NTC droplets in Fig. 5(a). This indicates that as the measurement of high-concentration droplets progresses, residual droplets continue to accumulate within the common transport channel, leading to severe contamination within the common transport channel.

[0177] In other words, this means that cleaning the common transport channel using conventional hydrophobic fluids cannot completely solve the contamination problem that must be critically handled in ddPCR. Furthermore, the experimental results in Figure 5 show that the hydrophobic fluid cleaning operation cannot prevent the continuous increase in contamination of the common transport channel.

[0178]

[0179] Meanwhile, to solve this problem, the applicant considered repeating the hydrophobic fluid washing operation several times between the droplet transfer operation of the first well and the droplet transfer operation of the second well, thereby repeatedly applying a force to the common transfer channel to push out the remaining droplets. However, repeating the washing operation using the hydrophobic fluid several times is undesirable because it increases the total time required to measure the droplets.

[0180] In addition, a method was considered to increase the amount of physical force pushing out residual droplets by increasing the positive pressure provided by the hydraulic supply unit (3000) during the hydrophobic fluid cleaning operation. However, if too much positive pressure is provided by the hydraulic supply unit (3000), there was a risk that the common movement channel would be damaged.

[0181] In addition, it was found that repeating the hydrophobic fluid cleaning operation multiple times or increasing the positive pressure of the hydraulic supply unit (3000) also ultimately cannot prevent the occurrence of contamination in the common moving channel and the increase in contamination due to repeated droplet measurements, as shown in the experimental results of FIG. 5.

[0182]

[0183] Accordingly, the applicant investigated the reason why residual droplets are not completely washed away by the hydrophobic fluid washing operation.

[0184] [Why hydrophobic fluid cleaning alone cannot solve the common transport channel contamination problem]

[0185]

[0186] As described above, in the conventional hydrophobic fluid cleaning operation, the hydrophobic fluid, to which hydraulic pressure is applied by the hydraulic supply unit (3000), pushes the residual droplets within the common movement channel to move to the fluid processing unit (7000). In other words, when the hydrophobic fluid to which hydraulic pressure is applied by the hydraulic supply unit (3000) comes close enough to meet the residual droplets, the residual droplets are hydrophilic, so there is insufficient attractive force to attract each other, and they do not mix with each other, and a boundary is formed between the hydrophobic fluid and the residual droplets. That is, due to the phase separation phenomenon in which the residual droplets do not want to mix with the hydrophobic fluid moving according to the hydraulic pressure of the hydraulic supply unit (3000), they continue to try to separate from the hydrophobic fluid and move in the direction of movement of the hydrophobic fluid, thereby moving to the fluid processing unit (7000). As the residual droplets move to the fluid processing unit (7000), the common movement channel is cleaned.

[0187] However, referring to Fig. 6(a), if we look at the path through which the droplet is transported, there are connecting parts (e.g., ports) that connect each part. Since the diameter of these connecting parts does not exactly match the diameter of the common transport channel, a step difference occurs between the channel within the common transport channel and the connecting parts.

[0188] The applicant confirmed that residual liquid droplets exist in the step difference between this connection part and the channel, and that the residual liquid droplets are not cleaned by utilizing only the property of separation between the hydrophobic fluid and the residual liquid droplets.

[0189] Unlike cases where the diameter of a channel through which a fluid moves remains constant or changes constantly along the length of the channel, when the diameter changes abruptly, such as with a step, a vortex is formed as the fluid moves near the step. As described above, the applicant presumes that the reason the residual droplet is not cleaned by utilizing only the property of separation between the hydrophobic fluid and the residual droplet at the step is due to the step.

[0190] Another reason why residual liquid droplets are not washed away is presumed to be due to surface tension. As shown in Fig. 6(b), when residual liquid droplets adhere to the inner wall of the valve or common moving channel, the surface tension of the residual liquid droplets is greater than the surface tension of the hydrophobic fluid, so the hydrophobic fluid cannot move the residual liquid droplets and flows around the residual liquid droplets, making it difficult for the residual liquid droplets to detach from the inner wall.

[0191] In addition, it was observed that as the droplets from the next well move through the common movement channel, they meet with residual droplets attached to the step or the inner wall of the common movement channel due to surface tension, dissolve each other, and move together with the residual droplets to the droplet detection channel (6000). Furthermore, it was observed that this results in false positive and / or false negative results.

[0192] Accordingly, the applicant devised a method to effectively clean residual liquid droplets attached to the inner wall of a step or common moving channel.

[0193]

[0194] [Method for washing by mixing residual droplets]

[0195] As mentioned above, the hydrophobic fluid cleaning operation cleans residual droplets by pushing them away with the physical pushing force of the hydrophobic fluid, but it was found that this is insufficient to completely clean the residual droplets.

[0196] Accordingly, the applicant has devised a method for dissolving (mixing) residual droplets in the common moving channel and proposes this in the present application. Specifically, the applicant proposes a method for washing by dissolving residual droplets in the common moving channel with a hydrophilic fluid that can be mixed with the droplets (hereinafter referred to as the hydrophilic fluid washing operation). Since droplets are hydrophilic, using a hydrophilic fluid causes the droplets and the hydrophilic fluid to dissolve, thereby enabling the washing of all residual droplets adhering to the common moving channel or existing on the steps.

[0197] In addition, since the hydrophilic fluid cleaning operation does not use physical force to clean residual droplets, there is no need to increase the number of cleaning operations or increase the positive pressure provided by the hydraulic supply unit (3000) to clean residual droplets. Therefore, the measurement time of the droplets is not increased, and no damage is caused to the common moving channel or the components of the droplet analysis device (10).

[0198]

[0199] [Problems when changing hydrophobic fluid cleaning operation to hydrophilic fluid cleaning operation]

[0200] Meanwhile, considering the solution described above, one can consider changing the existing hydrophobic fluid washing operation to a hydrophilic fluid washing operation. That is, residual droplets can be sufficiently removed by performing a hydrophilic fluid washing operation instead of a hydrophobic fluid washing operation. Therefore, one can consider a method of eliminating the hydrophobic fluid washing operation and performing only the hydrophilic fluid washing operation.

[0201] However, if only a hydrophilic fluid cleaning operation is performed without performing a hydrophobic fluid cleaning operation, the inside of the common transfer channel is filled with the hydrophilic fluid.

[0202] When the inside of the common transport channel is filled with a hydrophilic fluid and the droplet transport operation described in Fig. 3 is performed, the droplet moves through the common transport channel and comes into contact with the hydrophilic fluid, causing it to dissolve. Since the dissolved droplet is decomposed by the hydrophilic fluid, fluorescence detection becomes impossible. In other words, if a hydrophilic fluid washing operation is performed instead of a hydrophobic fluid washing operation, the droplet cannot be completely transported to the droplet detection channel. Consequently, fluorescence cannot be detected from the droplet.

[0203]

[0204] Accordingly, the present disclosure proposes a method for cleaning residual droplets by performing a hydrophilic fluid cleaning operation and a hydrophobic fluid cleaning operation together.

[0205] [Operation of the droplet analysis device (10) proposed for washing residual droplets]

[0206] FIG. 7 shows the operation of a droplet analysis device (10) for cleaning residual droplets proposed in the present disclosure.

[0207] According to FIG. 7, the droplet analysis device (10) performs a droplet transfer operation (S710), performs a hydrophilic fluid washing operation (S730), and then performs a hydrophobic fluid washing operation (S750). Then, the droplet analysis device (10) can perform a droplet transfer operation (S710). For example, the droplet analysis device (10) can perform a droplet transfer operation (S710) to transfer droplets from a first well to a droplet detection channel, perform a hydrophilic fluid washing operation (S730), and perform a hydrophobic fluid washing operation (S750). After that, the droplet analysis device (10) can perform a droplet transfer operation (S710) to transfer droplets from a second well, which is different from the first well, to a droplet detection channel, perform a hydrophilic fluid washing operation (S730), and then perform a hydrophobic fluid washing operation (S750). In this way, the droplet analysis device (10) can repeatedly perform a droplet transfer operation (S710) for transferring droplets from each well to a droplet detection channel, a hydrophilic fluid washing operation (S730), and a hydrophobic fluid washing operation (S750) until the droplet transfer operation (S710) of all wells included in the droplet storage unit is performed.

[0208] Here, the hydrophilic fluid washing operation (S730) must be performed between the hydrophobic fluid washing operation (S750) and the droplet transfer operation (S710). That is, the hydrophilic fluid washing operation (S730) must be performed before the hydrophobic fluid washing operation (S750). In other words, the hydrophobic fluid washing operation (S730) must not be performed between the hydrophobic fluid washing operation (S750) and the droplet transfer operation (S710) of the next well.

[0209] This is to allow the hydrophilic fluid filling the common transfer channel according to the hydrophilic fluid washing operation (S730) to be discharged to the fluid treatment plant by the hydrophobic fluid according to the hydrophobic fluid washing operation (S750). This is to prevent the droplet of the target well, which is the subject of fluorescence measurement, from being decomposed by the hydrophilic fluid filling the common transfer channel.

[0210] In other words, the purpose is to ensure that fluorescence measurements are performed properly by filling the common transport channel with a hydrophobic fluid through a hydrophobic fluid washing operation after washing the common transport channel with a hydrophilic fluid, thereby allowing the droplet in the target well to move through the common transport channel while maintaining its shape.

[0211] Meanwhile, the hydrophobic fluid washing operation (S750) requires supplying hydrophobic fluid to all channels through which the hydrophobic fluid and droplets can move within the droplet analysis device (10) for the droplet transfer operation (S710) of the next well. This is because if a hydrophilic fluid or air present in some of the channels through which the hydrophobic fluid and droplets must move within the droplet analysis device (10) flows into the common movement channel through which the droplets move, it interferes with the fluorescence detection of the droplets. Therefore, the hydrophobic fluid washing operation (S750) can control the droplet analysis device (10) to change the channel paths several times to supply hydrophobic fluid to each of the changed channel paths in order to supply hydrophobic fluid to all channels through which the hydrophobic fluid and droplets can move within the droplet analysis device (10).

[0212] On the other hand, the hydrophilic fluid washing operation (S730) is intended to remove residual droplets. Therefore, a hydrophilic fluid must be supplied to the common movement channel through which the droplets move. However, since droplets do not move through channels other than the common movement channel, no residual droplets remain. Therefore, supplying a hydrophilic fluid to channels other than the common movement channel results in wasted control operations of the droplet analysis device (10) and only increases the total time required for fluorescence measurement of the droplets.

[0213] Therefore, the hydrophilic fluid washing operation (S730) is preferably performed to minimize the operation of the droplet analysis device (10) such that the hydrophilic fluid is supplied to the common moving channel, but the supply of the hydrophilic fluid to channels other than the common channel is minimized.

[0214] Meanwhile, according to the operation of the droplet analysis device (10) proposed in the present disclosure, both a hydrophilic fluid washing operation and a hydrophobic fluid washing operation must be performed. Accordingly, the droplet analysis device (10) according to the present disclosure may additionally provide a hydrophilic fluid storage tank (e.g., water tank) for storing a hydrophilic fluid in addition to a hydrophobic fluid storage tank (e.g., oil tank) for storing a hydrophobic fluid.

[0215]

[0216] Hereinafter, in the present disclosure, various embodiments of a droplet analysis device (10) that performs the operations described in FIG. 7 and their operations will be examined.

[0217] [Example of a droplet analysis device (10) performing the operation of FIG. 7]

[0218] [1] Example 1

[0219] FIG. 8 shows the configuration of a droplet analysis device (10) according to Example 1. The droplet analysis device (10) according to Example 1 includes a controller (1000), a droplet suction device (4000), a droplet storage unit (9000), a hydrophilic fluid storage unit (8100), a hydrophobic fluid storage unit (8200), an X junction member (5400), a droplet detection channel (6000), a valve (5500), a first path control unit (5100), a second path control unit (5200), a third path control unit (5300), and a hydraulic supply unit (not shown). Additionally, the dotted lines in FIG. 8 represent channels to which the components of the droplet analysis device (10) are fluidly connected.

[0220] The controller (1000) can control the operation of the first path control unit (5100), the second path control unit (5200), the third path control unit (5300), the valve (5500), the droplet suction unit (4000), and the hydraulic supply unit (not shown).

[0221] Here, the X junction member (5400) may be a fitting having an X junction. Additionally, the X junction member (5400) may have a junction outlet, a junction center inlet, and at least two junction side inlets. The junction outlet of the X junction member (5400) may be fluidly connected to the detection inlet of the droplet detection channel (6000).

[0222] The first path control unit (5100) can selectively connect the droplet suction device (4000) and the junction center inlet of the X junction member. The second path control unit (5200) can selectively connect the hydrophilic fluid reservoir (8100) and at least two junction side inlets of the X junction member. The third path control unit (5300) can selectively connect the hydrophobic fluid reservoir (8200) and at least two junction side inlets of the X junction member. The valve (5500) can be fluidly connected to the detection outlet of the droplet detection channel (6000).

[0223] Meanwhile, the droplet analysis device (10) according to Example 1 may include a configuration not disclosed in FIG. 8. For example, at least one fluorescence detector (2000) may be arranged on one side of the droplet detection channel (6000).

[0224] FIG. 9 shows the droplet analysis device (10) according to Example 1 performing the droplet transfer operation (S710) of FIG. 7. Prior to the droplet transfer operation (S710) of the droplet analysis device (10) according to Example 1, all channels indicated by the dotted line in FIG. 8 are filled with a hydrophobic fluid.

[0225] Referring to FIG. 9, the controller (1000) positions the droplet suction device (4000) in the first well of the droplet storage unit (9000) and closes the valve (5500) (OFF). The controller (1000) controls the first path control unit (5100) so that the droplet suction device (4000) and the junction center inlet of the X-junction member (5400) are fluidly connected. The controller (1000) controls the third path control unit (5300) so that at least two junction side inlets of the X-junction member (5400) are fluidly connected to the third path control unit (5300). The controller (1000) controls the hydraulic supply unit (not shown) to apply negative pressure to the third path control unit (5300). Under the control of the controller (1000) described above, the droplet moves from the first well through the droplet suction device (4000) to the first channel between the first path control unit (5100) and the junction center inlet of the X junction member (5400). The operation of the controller (1000) described above is called the first droplet transfer operation.

[0226] Referring to FIG. 10, the controller (1000) opens the valve (5500) (ON). The controller (1000) controls the first path control unit (5100) so that the junction center inlet of the droplet suction unit (4000) and the X-junction member (5400) are not fluidly connected. The controller (1000) controls the hydraulic supply unit (not shown) to apply positive pressure to the first path control unit (5100). Additionally, the controller (1000) may control the third path control unit (5300) so that at least two junction side inlets of the X-junction member (5400) are not fluidly connected to the third path control unit (5300).

[0227] According to the control of the controller (1000) described above, a droplet located in the first channel between the first path control unit (5100) and the junction center inlet of the X junction member (5400) can pass through the droplet detection channel (6000) via the outlet of the X junction member (5400) and the detection inlet of the droplet detection channel (6000). The operation of the controller (1000) described above is called the second droplet transfer operation.

[0228] In FIGS. 9 and 10, droplets moved through channels connecting the droplet suction device (4000), the first path control unit (5100), the junction center inlet and junction outlet of the X junction member (5400), and the droplet detection channel (6000). Therefore, in Example 1, the channels connecting the droplet suction device (4000), the first path control unit (5100), the junction center inlet and junction outlet of the X junction member (5400), and the droplet detection channel (6000) become common movement channels.

[0229] FIGS. 11 and 12 illustrate a hydrophilic fluid washing operation for washing residual droplets remaining in the common moving channel according to Example 1.

[0230] Referring to FIGS. 11 and 12, the controller (1000) can move the droplet suction device (4000) to a designated location. For example, the designated location is a location spaced apart from the droplet storage unit (9000), and when the droplet suction device (4000) is located at the designated location, the tip of the droplet suction device (4000) may be directed toward a place other than the droplet storage unit (9000). Meanwhile, the controller (1000) may also move the droplet suction device (4000) to a designated location during a droplet transfer operation. For example, the controller (1000) may move the droplet suction device (4000) to the designated location after the first droplet transfer operation is performed and before the hydrophilic fluid cleaning operation begins. This is to prevent residual droplets and / or fluid discharged from the droplet suction device (4000) during the cleaning operation from entering the droplet storage unit (9000).

[0231] The controller (1000) can close (OFF) the valve (5500). For example, the controller (1000) may keep the valve (5500) closed while performing a hydrophilic fluid cleaning operation. Alternatively, the controller (1000) may close the valve (5500) only during a portion of the time interval during which the hydrophilic fluid cleaning operation is performed, and keep it open during the remainder of the time interval. For example, the controller (1000) may close the valve (5500) while starting the hydrophilic fluid cleaning operation and open the valve (5500) after a first time has elapsed, or it may open the valve (5500) when starting the hydrophilic fluid cleaning operation and close the valve (5500) after a second time has elapsed. Here, the first time and the second time may be the same or different.

[0232] The controller (1000) controls the first path control unit (5100) so that the junction center inlet of the droplet suction unit (4000) and the X-junction member (5400) is fluidically connected. The controller (1000) controls the second path control unit (5200) so that at least two junction side inlets of the hydrophilic fluid reservoir (8100) and the X-junction member (5400) are fluidly connected. The controller (1000) controls the hydraulic supply unit (not shown) to apply positive pressure to the second path control unit (5200). During the time interval when the valve (5500) is closed, as can be seen in FIG. 11, the hydrophilic fluid is introduced into at least two junction side inlets of the X-junction member (5400) under the control of the controller (1000) described above, and can pass through the X-junction member (5400) and the droplet suction device (4000). Additionally, during the time interval when the valve (5500) is open, as can be seen in FIG. 12, the hydrophilic fluid is introduced into at least two junction side inlets of the X-junction member (5400) under the control of the controller (1000) described above, and can pass through the droplet detection channel (6000) while passing through the X-junction member (5400) and the droplet suction device (4000).

[0233] Accordingly, residual droplets remaining in the channel between the X-junction member (5400) and the droplet suction device (4000) in the common moving channel can be dissolved in a hydrophilic fluid and washed. The operation of the controller (1000) described above is called a hydrophilic fluid washing operation.

[0234] FIGS. 13 to 15 illustrate a hydrophobic fluid washing operation for washing residual droplets remaining in the common moving channel. The hydrophobic fluid washing operation includes a first hydrophobic fluid washing operation and a second hydrophobic fluid washing operation.

[0235] The first hydrophobic fluid cleaning operation may be either hydrophobic fluid cleaning operation 'A' as in FIG. 13 or hydrophobic fluid cleaning operation 'B' as in FIG. 14.

[0236] Referring to FIG. 13, to examine hydrophobic fluid cleaning operation 'A', the controller (1000) opens (ON) the valve (5500). The controller (1000) controls the third path control unit (5300) so that at least two junction side inlets of the hydrophobic fluid reservoir (8200) and the X-junction member (5400) are fluidly connected. The controller (1000) controls the hydraulic supply unit (not shown) to apply positive pressure to the third path control unit (5300). Under the control of the controller (1000) described above, the hydrophobic fluid passes through the droplet detection channel (6000) via at least two junction side inlets of the X-junction member (5400). Accordingly, the X-junction member (5400) to the droplet detection channel (6000) can be cleaned by the hydrophobic fluid. Additionally, during hydrophobic fluid cleaning operation 'A', the hydrophilic fluid filled from the third path control unit (5300) to the X-junction member (5400) passes through the droplet detection channel (6000) by the positive pressure applied to the third path control unit (5300) by the hydraulic supply unit (not shown), and the remaining droplet from the X-junction member (5400) to the droplet detection channel (6000) can be dissolved and cleaned.

[0237] Meanwhile, hydrophobic fluid cleaning operation 'B' may be performed instead of hydrophobic fluid cleaning operation 'A'. Referring to FIG. 14, the controller (1000) opens (ON) the valve (5500). The controller (1000) controls the first path control unit (5100) so that the hydrophobic fluid reservoir (8200) and the junction center inlet of the X-junction member (5400) are fluidly connected. The controller (1000) controls the hydraulic supply unit (not shown) to apply positive pressure to the first path control unit (5100). Under the control of the controller (1000) described above, the hydrophobic fluid passes through the junction center inlet of the X-junction member (5400) and through the droplet detection channel (6000). At this time, the hydrophilic fluid filled from the first path control unit (5100) to the X-junction member (5400) passes through the droplet detection channel (6000) by the positive pressure applied to the first path control unit (5300) by the hydraulic supply unit (not shown), and the remaining droplet from the X-junction member (5400) to the droplet detection channel (6000) can be dissolved and washed.

[0238] After hydrophobic fluid cleaning operation 'A' or hydrophobic fluid cleaning operation 'B' is performed, a second hydrophobic fluid cleaning operation may be performed. Referring to FIG. 15, to perform the second hydrophobic fluid cleaning operation, the controller (1000) may close the valve (5500) (OFF). However, as mentioned in the description of the hydrophilic fluid cleaning operation above, the controller (1000) may close the valve (5500) only during a portion of the time interval in which the second hydrophobic fluid cleaning operation is performed, and open it during the remaining interval. The controller (1000) controls the first path control unit (5100) so that the junction center inlet of the droplet suction device (4000) and the X-junction member (5400) are fluidly connected. The controller (1000) controls the third path control unit (5300) so that at least two junction side inlets of the hydrophobic fluid reservoir (8100) and the X-junction member (5400) are fluidly connected. The controller (1000) controls the hydraulic supply unit (not shown) to apply positive pressure to the third path control unit (5300). Under the control of the controller (1000) described above, the hydrophobic fluid flows into at least two junction side inlets of the X-junction member (5400), passes through the junction center inlet of the X-junction member (5400), and passes through the droplet suction unit (4000).

[0239] According to the first hydrophobic fluid washing operation (i.e., hydrophobic fluid washing operation 'A' or hydrophobic fluid washing operation 'B') and the second hydrophobic fluid washing operation, the hydrophobic fluid passes through and fills all channels through which the hydrophobic fluid and droplets can move.

[0240] FIGS. 16 and 17 illustrate a droplet transfer operation in which a droplet is transferred again after a hydrophobic fluid washing operation. The controller (1000) positions the droplet suction device (4000) in the second well of the droplet storage unit (9000). At this time, the second well is a different well from the first well where the droplet suction device (4000) is located in FIGS. 9 and 10. Subsequently, the controller (1000) performs the first droplet transfer operation and the second droplet transfer operation described through FIGS. 9 and 10 in the same manner.

[0241] Accordingly, the droplets of the second well are transported along the same common transport channel as the droplets of the first well, and some of the droplets of the second well remain in the common transport channel. Additionally, the controller (1000) performs the hydrophilic fluid washing operation and the hydrophobic fluid washing operation described above, and the droplets remaining in the common transport channel can be washed.

[0242] [2] Example 2

[0243] FIG. 18 shows the configuration of a droplet analysis device (10) according to Example 2. The droplet analysis device (10) according to Example 2 does not include the X junction member (5400) among the configurations of the droplet analysis device according to Example 1. On the other hand, the droplet detection channel (6000) may have a central inlet, at least two side inlets, and an outlet. Additionally, the droplet detection channel (6000) may have an X junction and may include a central inlet channel extended between the central inlet and the X junction, at least two side channels extended between each of the at least two side inlets and the X junction, and an outlet channel extended between the outlet and the X junction.

[0244] Meanwhile, for example, the droplet detection channel (6000) may be configured in the form of a plate. For example, a central channel, at least two side channels, and an outlet channel may be formed within an optically transparent plate, and a central inlet, at least two side inlets, and an outlet may be formed on the outer surface of the plate.

[0245] Except for the above-described points, the droplet analysis device (10) according to Embodiment 2 may have the same components and fluid connection relationships between components as the droplet analysis device according to Embodiment 1. Meanwhile, accordingly, in Embodiment 2, the first path control unit (5100) selectively connects the central inlet of the droplet detection channel (6000) to the droplet suction device (4000), and the second path control unit (5200) selectively connects the hydrophilic fluid reservoir (8100) to at least two side inlets of the droplet detection channel (6000). Additionally, the third path control unit (5300) selectively connects the hydrophobic fluid reservoir (8200) to at least two side inlets of the droplet detection channel (6000).

[0246] In Example 2 as well, all channels indicated by the dotted line in FIG. 18 are filled with a hydrophobic fluid prior to the droplet transfer operation (S710) of the droplet analysis device (10).

[0247] FIGS. 19 and 20 illustrate a droplet analysis device (10) according to Example 2 performing a droplet transfer operation (S710). FIG. 19 illustrates a droplet analysis device (10) according to Example 2 performing a first droplet transfer operation. Referring to FIG. 19, the controller (1000) controls the droplet suction device (4000), valve (5500), first path control unit (5100), third path control unit (5300), and hydraulic supply unit (not shown) in the same manner as described in FIG. 9 and the first droplet transfer operation of Example 1. However, the difference between the first droplet transfer operation of Example 1 and the first droplet transfer operation of Example 2 is, with reference to FIG. 19, that the controller (1000) controls the first path control unit (5100) so that the droplet suction device (4000) and the central inlet of the droplet detection channel (6000) are fluidly connected, and the controller (1000) controls the third path control unit (5300) so that at least two side inlets of the droplet detection channel (6000) are fluidly connected to the third path control unit (5300). Additionally, accordingly, the droplet moves from the first well through the droplet suction device (4000) to the first channel between the first path control unit (5100) and the central inlet of the droplet detection channel (6000).

[0248] FIG. 20 shows a droplet analysis device (10) according to Example 2 performing a second droplet transfer operation. Referring to FIG. 20, the controller (1000) can control the droplet suction device (4000), valve (5500), first path control unit (5100), third path control unit (5300), and hydraulic supply unit (not shown) in the same way as described in FIG. 10 and the second droplet transfer operation of Example 1. However, the difference between the second droplet transfer operation of Example 1 and the second droplet transfer operation of Example 2 is, with reference to FIG. 20, that the controller (1000) controls the first path control unit (5100) so that the central inlet of the droplet suction device (4000) and the droplet detection channel (6000) are fluidly connected, and controls the third path control unit (5300) so that at least two side inlets of the droplet detection channel (6000) are not fluidly connected to the third path control unit (5300). Additionally, accordingly, the droplet passes through the droplet detection channel (6000).

[0249] As described above, in Example 2, the central inlet channel and outlet channel of the droplet suction device (4000), the first path control unit (5100), and the droplet detection channel (6000) become a common moving channel.

[0250] FIGS. 21 and 22 illustrate a hydrophilic fluid washing operation for washing residual droplets remaining in the common moving channel according to Example 2.

[0251] Referring to FIGS. 21 and 22, the controller (1000) can control the liquid dropper (4000), valve (5500), first path control unit (5100), and second path control unit (5200) in the same manner as described in FIGS. 11 and 12 as well as the hydrophilic fluid washing operation according to Example 1.

[0252] However, the difference between the hydrophilic fluid washing operation in Example 2 and the hydrophilic fluid washing operation in Example 1 is that the controller (1000) controls the first path control unit (5100) so that the central inlet of the droplet suction device (4000) and the droplet detection channel (6000) are fluidly connected, and controls the second path control unit (5200) so that at least two side inlets of the hydrophilic fluid reservoir (8100) and the droplet detection channel (6000) are fluidly connected. Accordingly, residual droplets remaining in the channel between the central inlet channel of the droplet detection channel (6000) and the droplet suction device (4000) among the common moving channels can be dissolved in the hydrophilic fluid and washed.

[0253] FIGS. 23 to 25 illustrate a hydrophobic fluid cleaning operation. FIG. 23 illustrates a hydrophobic fluid cleaning operation 'A'. Referring to FIG. 23, the controller (1000) can control the valve (5500) and the third path control unit (5300) in the same way as the hydrophobic fluid cleaning operation 'A' according to Embodiment 1. However, the difference between the hydrophobic fluid cleaning operation 'A' of Embodiment 1 and the hydrophobic fluid cleaning operation 'A' of Embodiment 2 is that the controller (1000) controls the third path control unit (5300) so that at least two side inlets of the hydrophobic fluid reservoir (8200) and the droplet detection channel (6000) are fluidly connected. Additionally, during hydrophobic fluid cleaning operation 'A', the hydrophilic fluid that was filled from the third path control unit (5300) to the X junction of the droplet detection channel (6000) passes through the droplet detection channel (6000) by the positive pressure applied to the third path control unit (5300) by the hydraulic supply unit (not shown), and the remaining droplet in the outlet channel of the droplet detection channel (6000) can be dissolved and cleaned.

[0254] Meanwhile, in Example 2, hydrophobic fluid cleaning operation 'B' may be performed instead of hydrophobic fluid cleaning operation 'A'. Referring to FIG. 24, the controller (1000) can control the valve (5500) and the first path control unit (5100) in the same way as the hydrophobic fluid cleaning operation 'B' of Example 1. However, the difference between the hydrophobic fluid cleaning operation 'B' of Example 1 and the hydrophobic fluid cleaning operation 'B' of Example 2 is that the controller (1000) controls the first path control unit (5100) so that the hydrophobic fluid reservoir (8200) and the central inlet of the droplet detection channel (6000) are fluidly connected. At this time, the hydrophilic fluid that was filled from the first path control unit (5100) to the X-junction of the droplet detection channel (6000) passes through the droplet detection channel (6000) by the positive pressure applied to the first path control unit (5300) by the hydraulic supply unit (not shown), and the remaining droplet up to the outlet channel of the droplet detection channel (6000) can be dissolved and washed.

[0255] After hydrophobic fluid cleaning operation 'A' or hydrophobic fluid cleaning operation 'B' is performed, a second hydrophobic fluid cleaning operation may be performed. Referring to FIG. 25, the controller (1000) can control the valve (5500), the first path control unit (5100), and the third path control unit in the same way as the second hydrophobic fluid cleaning operation of Embodiment 1. However, the difference between the second hydrophobic fluid cleaning operation of Embodiment 1 and the second hydrophobic fluid cleaning operation of Embodiment 2 is that the controller (1000) controls the first path control unit (5100) so that the central inlet of the droplet suction device (4000) and the droplet detection channel (6000) are fluidly connected, and the controller (1000) controls the third path control unit (5300) so that at least two side inlets of the hydrophobic fluid reservoir (8200) and the droplet detection channel (6000) are fluidly connected.

[0256] Under the control of the controller (1000) described above, a hydrophobic fluid flows into at least two side inlets of the droplet detection channel (6000), passes through the central inlet of the droplet detection channel (6000), and passes through the droplet suction device (4000).

[0257] According to Example 2, the hydrophobic fluid passes through and fills all channels through which the hydrophobic fluid and droplets can move according to the first hydrophobic fluid washing operation (i.e., hydrophobic fluid washing operation 'A' or hydrophobic fluid washing operation 'B') and the second hydrophobic fluid washing operation.

[0258] FIGS. 26 and 27 illustrate a droplet transfer operation in which a droplet is transferred again after a hydrophobic fluid washing operation. The controller (1000) positions the droplet suction device (4000) in the second well of the droplet storage unit (9000). At this time, the second well is a different well from the first well where the droplet suction device (4000) is located in FIGS. 19 and 20. Subsequently, the controller (1000) performs the first droplet transfer operation and the second droplet transfer operation described through FIGS. 19 and 20 in the same manner.

[0259] Accordingly, the droplets of the second well are transported along the same common transport channel as the droplets of the first well, and some of the droplets of the second well remain in the common transport channel. Additionally, the controller (1000) performs the hydrophilic fluid washing operation and the hydrophobic fluid washing operation described above, and the droplets remaining in the common transport channel can be washed.

[0260] [3] Example 3

[0261] FIG. 28 shows the configuration of a droplet analysis device (10) according to Example 3. The droplet analysis device (10) according to Example 3 is identical in configuration to the droplet analysis device according to Example 1, except that a syringe pump (3100) is used as the hydraulic supply unit (3000). Referring to FIG. 18, the syringe pump is optionally connected to a hydrophilic fluid reservoir (8100) or a hydrophobic fluid reservoir (8200). That is, the syringe pump (3100) can load a hydrophilic fluid from the hydrophilic fluid reservoir (8100) or load a hydrophobic fluid from the hydrophobic fluid reservoir (8200). Additionally, depending on the operation of a valve mounted on or fluidly connected to the syringe pump, the hydrophilic fluid or hydrophobic fluid loaded into the syringe pump may be selectively provided to any one of the first path control unit (5100), the second path control unit (5200), and the third path control unit (5300).

[0262] In Example 3, an additional operation in which the cylinder pump (3100) loads a hydrophilic fluid or a hydrophobic fluid may be added to each operation described in Example 1.

[0263] FIGS. 29 to 31 show that a droplet analysis device (10) according to Example 3 performs a droplet transfer operation (S710).

[0264] Referring to FIG. 29, the controller (1000) controls the syringe pump (3100) to load hydrophobic fluid from the hydrophobic fluid reservoir (8200) before the first droplet transfer operation is performed. The controller (1000) then controls the syringe pump (3100) to provide hydraulic pressure so that the loaded hydrophobic fluid fills all channels indicated by the dotted line in FIG. 28. Referring to FIG. 30, the controller (1000) performs the first droplet transfer operation by controlling the valve (5500), the first path control unit (5100), and the third path control unit (5300) in the same manner as the first droplet transfer operation according to Embodiment 1. However, the difference between the first droplet transfer operation according to Example 1 and the first droplet transfer operation according to Example 3 is that the controller (1000) controls the third path control unit (5300) so that at least two junction side inlets of the X-junction member (5400) are fluidly connected to the syringe pump (3100), and the controller (1000) controls the syringe pump (3100) to apply negative pressure to the third path control unit (5300).

[0265] Referring to FIG. 31, the controller (1000) performs a second droplet transfer operation by controlling the valve (5500) and the first path control unit (5100) in the same manner as the second droplet transfer operation according to Embodiment 1. However, the difference between the second droplet transfer operation according to Embodiment 1 and the second droplet transfer operation according to Embodiment 3 is that the controller (1000) controls the first path control unit (5300) so that the junction center inlet of the X-junction member (5400) is fluidically connected to the syringe pump (3100), and the controller (1000) controls the syringe pump (3100) to apply positive pressure to the first path control unit (5100).

[0266] Except for the differences mentioned above, the path along which the droplet moves and the common movement channel are defined in the same way as in Example 1.

[0267] The controller (1000) controls the syringe pump (3100) to load the hydrophilic fluid from the hydrophilic fluid reservoir (8100) before performing the hydrophilic fluid washing operation as can be seen in FIG. 32.

[0268] Referring to FIGS. 33 to 34, the controller (1000) can perform a hydrophilic fluid cleaning operation in the same way as the hydrophilic fluid cleaning operation according to Example 1. However, the difference between the hydrophilic fluid cleaning operation according to Example 1 and the hydrophilic fluid cleaning operation according to Example 3 is that the controller (1000) controls the first path control unit (5300) so that the junction center inlet of the X-junction member (5400) is fluidly connected to the syringe pump (3000), the controller (1000) controls the second path control unit (5200) so that at least two junction side inlets of the syringe pump (3100) and the X-junction member (5400) are fluidly connected, and the controller (1000) controls the syringe pump (3000) so that the syringe pump (3100) applies positive pressure to the second path control unit (5200).

[0269] The controller (1000) controls the syringe pump (3100) to load hydrophobic fluid from the hydrophobic fluid reservoir (8200) before performing the first hydrophobic fluid washing operation as can be seen in FIG. 35.

[0270] Referring to FIG. 36, the controller (1000) can perform hydrophobic fluid cleaning operation 'A' in the same manner as the hydrophobic fluid cleaning operation 'A' according to Embodiment 1. However, the difference between the hydrophobic fluid cleaning operation 'A' according to Embodiment 1 and the hydrophobic fluid cleaning operation 'A' according to Embodiment 3 is that the controller (1000) controls the third path control unit (5300) so that at least two junction side inlets of the syringe pump (3100) and the X junction member (5400) are fluidly connected, and the controller (1000) controls the syringe pump (3000) so that the syringe pump (3000) applies positive pressure to the third path control unit (5300).

[0271] Referring to FIG. 37, the controller (1000) may perform hydrophobic fluid cleaning operation 'B' instead of hydrophobic fluid cleaning operation 'A'. Additionally, the controller (1000) may perform hydrophobic fluid cleaning operation 'B' in the same manner as hydrophobic fluid cleaning operation 'B' according to Embodiment 1. However, the difference between the hydrophobic fluid cleaning operation 'B' according to Embodiment 1 and the hydrophobic fluid cleaning operation 'B' according to Embodiment 3 is that the controller (1000) controls the first path control unit (5100) so that the syringe pump (3100) and the junction center inlet of the X junction member (5400) are fluidly connected, and the controller (1000) controls the syringe pump (3000) so that the syringe pump (3000) applies positive pressure to the first path control unit (5100).

[0272] The principle of the hydrophilic fluid washing the remaining droplets of the droplet detection channel (6000) according to the above-described hydrophobic fluid washing operation 'A' or hydrophobic fluid washing operation 'B' is the same as that described in Example 1.

[0273] Meanwhile, after hydrophobic fluid cleaning operation 'A' or hydrophobic fluid cleaning operation 'B' is performed, a second hydrophobic fluid cleaning operation may be performed. Referring to FIG. 38, the controller (1000) may perform the second hydrophobic fluid cleaning operation in the same manner as the second hydrophobic fluid cleaning operation according to Embodiment 1. However, the difference between the second hydrophobic fluid cleaning operation according to Embodiment 1 and the second hydrophobic fluid cleaning operation according to Embodiment 3 is that the controller (1000) controls the first path control unit (5100) so that the junction center inlet of the droplet suction device (4000) and the X junction member (5400) is fluidly connected, and the controller (1000) controls the third path control unit (5100) so that at least two junction side inlets of the syringe pump (3100) and the X junction member (5400) are fluidly connected. In addition, there is a difference in that the controller (1000) controls the syringe pump (3000) so that the syringe pump (3000) applies positive pressure to the third path control unit (5100).

[0274] According to the first hydrophobic fluid washing operation (i.e., hydrophobic fluid washing operation 'A' or hydrophobic fluid washing operation 'B') and the second hydrophobic fluid washing operation, the hydrophobic fluid passes through and fills all channels through which the hydrophobic fluid and droplets can move.

[0275] Referring to FIG. 39, the controller (1000) can control the syringe pump (3100) to load hydrophobic fluid from the hydrophobic fluid reservoir (8200) before performing a droplet transfer operation to measure the droplet of the second well.

[0276] After that, the controller (1000) can perform the first droplet transfer operation of FIG. 40 and the second droplet transfer operation of FIG. 41. The controller (1000) can position the droplet suction device (4000) in the second well of the droplet storage unit (9000) and then perform the first droplet transfer operation and the second droplet transfer operation in the same manner as FIG. 30 and FIG. 31.

[0277] Accordingly, the droplets of the second well are transported along the same common transport channel as the droplets of the first well, and some of the droplets of the second well remain in the common transport channel. Additionally, the controller (1000) performs the hydrophilic fluid washing operation and the hydrophobic fluid washing operation described above, and the droplets remaining in the common transport channel can be washed.

[0278] [4] Example 4

[0279] FIG. 42 shows the configuration of a droplet analysis device (10) according to Example 4. The droplet analysis device (10) according to Example 4 includes a controller (1000), a droplet suction device (4000), a droplet storage unit (9000), a hydrophilic fluid storage unit (8100), a hydrophobic fluid storage unit (8200), an X junction member (5400), a droplet detection channel (6000), a valve (5500), a T junction member (5700), a droplet processing unit (7000), a multi-port valve (5600), a first syringe pump (3100), and a second syringe pump (3200). Additionally, the dotted lines in FIG. 42 represent channels to which the components of the droplet analysis device (10) are fluidly connected.

[0280] The controller (1000) can control the operation of the first syringe pump (3100), the second syringe pump (3200), the valve (5500), the multi-port valve (5600), and the droplet suction device (4000). Additionally, the X junction member (5400) is a fitting having an X junction and may have a junction outlet, a junction center inlet, and at least two junction side inlets. The junction outlet of the X junction member (5400) may be fluidly connected to the detection inlet of the droplet detection channel (6000).

[0281] Additionally, the T junction member (5700) is a fitting having a T junction and may have a junction inlet and at least two junction side outlets. Here, each of the at least two junction side outlets may be fluidly connected to each of the at least two junction side inlets of the X junction member (5400).

[0282] The multi-port valve (5600) optionally connects the junction center inlet of the X junction member (5400) to the droplet suction device (4000) or the first syringe pump (3100). Additionally, when the multi-port valve (5600) connects the junction center inlet of the X junction member (5400) to the droplet suction device (4000), the multi-port valve (5600) fluidly connects the junction center inlet of the X junction member to the internal channel of the droplet suction device (4000), while fluidly connecting the first syringe pump (3100) to the external channel of the droplet suction device (4000).

[0283] The first syringe pump (3100) may be connected to a hydrophobic fluid storage unit (8200). Additionally, the second syringe pump (3200) may be optionally connected to either the hydrophobic fluid storage unit (8200) or the hydrophilic fluid storage unit (8100). Furthermore, the second syringe pump (3200) is fluidically connected to at least two junction side inlets of the X junction member (5400) via the T junction member (5700).

[0284] The descriptions of the other components are the same as those above, so they are omitted.

[0285] Additionally, the droplet analysis device (10) according to Example 4 may include a configuration not disclosed in FIG. 42. For example, at least one fluorescence detector (2000) may be arranged on one side of the droplet detection channel (6000). At this time, since the operation and function of the at least one fluorescence detector (2000) are the same as described above, a description will be omitted.

[0286]

[0287] FIGS. 43 to 45 are for explaining the droplet transfer operation of the droplet analysis device (10) according to Example 4. Referring to FIG. 43, the controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that both the first syringe pump (3100) and the second syringe pump (3200) are connected to the hydrophobic fluid storage unit (8200), and controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200). And the controller (1000) controls the syringe pump (3100) so that the syringe pump (3100) provides hydraulic pressure so that the loaded hydrophobic fluid fills all the channels indicated by the dotted line in FIG. 42.

[0288] Referring to FIG. 44, the controller (1000) performs a first droplet transfer operation. For example, the controller (1000) closes (OFF) the valve (5500) and positions the droplet suction device (4000) in the first well of the droplet storage unit (9000). Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the junction center inlet of the X-electric member (5400) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0289] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that hydrophobic fluid is transferred to the external channel of the droplet suction device (4000). The controller (1000) controls the second syringe pump (3200) to apply negative pressure so that droplets of the first well are positioned between the multi-port valve (5600) and the junction center inlet of the X junction member (5400).

[0290] Referring to FIG. 45, after performing the first droplet transfer operation, the controller (1000) performs the second droplet transfer operation. For example, the controller (1000) opens the valve (5500) and controls the multi-port valve (5600) so that the junction center inlet of the first syringe pump (3100) and the X-junction member (5400) is fluidly connected. At this time, the controller (1000) can move the droplet suction device (4000) to a designated position. At this time, the designated position is a position where the droplet or fluid emitted from the internal channel of the droplet suction device (4000) is transferred to the droplet processing unit (7000). However, the controller (1000) does not necessarily move the droplet suction device (4000) to a designated position during the second droplet transfer operation, and the controller (1000) may move the droplet suction device (4000) to a designated position before the second droplet transfer operation and the hydrophilic fluid cleaning of FIGS. 47 to 48 described later.

[0291] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that droplets located between the multi-port valve (5600) and the junction center inlet of the X junction member (5400) pass through the droplet detection channel (6000). At this time, the controller (1000) controls the second syringe pump (3200) to apply positive pressure so that droplets passing through the X junction member (5400) can meet the hydrophobic fluid injected into at least two side inlets of the X junction member (5400). Droplets that meet the hydrophobic fluid in the X junction member (5400) are sufficiently separated from each other.

[0292] While droplets pass through the droplet detection channel (6000), the controller (1000) controls at least one fluorescence detector to detect fluorescence from the droplets. The droplets that have passed through the droplet detection channel (6000) are transferred to the droplet processing unit (7000) via the valve (5500).

[0293] Referring to the first droplet transfer operation and the second droplet transfer operation, the droplets of the first well pass through the internal channel of the droplet suction device (4000), the multi-port valve (5600), the X-junction member (5400), and the droplet detection channel (6000). The path along which the droplets of the first well have traveled is called the common travel channel, and some of the droplets of the first well remain in the common travel channel.

[0294] Referring to FIG. 46, after the first and second droplet transfer operations for the first well are performed, the controller (1000) controls the second syringe pump (3100) so that the second syringe pump (3100) and the hydrophilic fluid storage unit (8100) are fluidly connected. Additionally, the controller (1000) controls the first syringe pump (3100) so that the first syringe pump (3100) loads hydrophobic fluid from the hydrophobic fluid storage unit (8200), and controls the second syringe pump (3200) so that the second syringe pump (3200) loads hydrophilic fluid from the hydrophilic fluid storage unit (8100).

[0295] Referring to FIGS. 47 and 48, the controller (1000) can perform a hydrophilic fluid cleaning operation. For example, the controller (1000) can close (OFF) the valve (5500). For example, the controller (1000) may keep the valve (5500) closed while performing the hydrophilic fluid cleaning operation. Alternatively, the controller (1000) may close the valve (5500) only during a portion of the time interval in which the hydrophilic fluid cleaning operation is performed, and open it during the remaining interval. For example, the controller (1000) may close the valve (5500) while starting the hydrophilic fluid cleaning operation and open the valve (5500) after a first time has elapsed, or it may open the valve (5500) when starting the hydrophilic fluid cleaning operation and close the valve (5500) after a second time has elapsed. Here, the first time and the second time may be the same or different. At this time, the droplet suction device (4000) is positioned at the designated location. Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the junction center inlet of the X-electric member (5400) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0296] The controller (1000) can control the first syringe pump (3100) to apply positive pressure so that the hydrophobic fluid is transferred to the external channel of the droplet suction device (4000), but this may be omitted. During the time interval when the valve (5500) is closed, as can be seen in FIG. 47, the controller (1000) can control the second syringe pump (3200) to apply positive pressure so that the hydrophilic fluid is transferred to the droplet processing unit (7000) through the X junction member (5400), the multi-port valve (5600), and the internal channel of the droplet suction device (4000). Additionally, during the time interval when the valve (5500) is open, as can be seen in FIG. 48, the controller (1000) described above controls the second syringe pump (3200) to apply positive pressure, so that a hydrophilic fluid flows into at least two junction side inlets of the X-junction member (5400), passes through the X-junction member (5400), passes through the droplet suction device (4000), and also passes through the droplet detection channel (6000).

[0297] Accordingly, residual droplets in a portion of the common moving channel leading to the internal channel of the X junction member (5400), multi-port valve (5600), and droplet suction device (4000) are washed by a hydrophilic fluid.

[0298] After the hydrophilic fluid washing operation, the controller (1000) controls the second syringe pump (3200) so that the second syringe pump (3200) and the hydrophobic fluid storage unit (8200) are connected as shown in FIG. 49.

[0299] Additionally, the controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200).

[0300] Referring to FIG. 50, the controller (1000) performs a first hydrophobic fluid cleaning operation. For example, the controller (1000) opens the valve (5500) and controls the multiport valve (5600) so that the first syringe pump (3100) and the junction center inlet of the X-junction member (5400) are fluidly connected. The controller (1000) controls the first syringe pump (3100) so that the first syringe pump (3100) applies positive pressure to the junction center inlet of the X-junction member (5400). The application of positive pressure by the first syringe pump (3100) causes the hydrophilic fluid, which was filled between the multi-port valve (5600) and the junction center inlet of the X junction member (5400) during the hydrophilic fluid washing operation, to pass through the droplet detection channel (6000) and move to the droplet processing unit (7000). Accordingly, the remaining droplets in the droplet detection channel (6000) that were not washed during the hydrophilic fluid washing operation are washed by the hydrophilic fluid.

[0301] Additionally, after the remaining droplets are washed by a hydrophilic fluid, the channel connecting the first syringe pump (3100), the multi-port valve (5600), the X-junction member (5400), and the droplet detection channel (6000) is washed once more by a hydrophobic fluid. Meanwhile, the controller (1000) controls the second syringe pump (3200) so that the second syringe pump (3200) applies positive pressure to at least two junction-side inlets of the X-junction member (5400). Accordingly, the channel leading to the second syringe pump (3200), the X-junction member (5400), and the droplet detection channel (6000) is washed by a hydrophobic fluid.

[0302] Referring to FIG. 51, the controller (1000) performs a second hydrophobic fluid cleaning operation. For example, the controller (1000) can close the valve (5500) (OFF). However, as mentioned in the description of the hydrophilic fluid cleaning operation above, the controller (1000) may close the valve (5500) only during a portion of the time interval in which the second hydrophobic fluid cleaning operation is performed, and open it during the remaining interval. At this time, the droplet suction device (4000) is located at the designated position. Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the junction center inlet of the X-electric member (5400) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0303] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that hydrophobic fluids are transferred to the external channel of the droplet suction device (4000). The controller (1000) controls the second syringe pump (3200) to apply positive pressure so that hydrophobic fluids can be transferred to the droplet processing unit (7000) through the X junction member (5400), the multi-port valve (5600), and the internal channel of the droplet suction device (4000).

[0304] When the hydrophobic fluid washing operation is completed, the controller (1000) performs a droplet transfer operation to measure droplets in the second well that are different from the first well. Referring to FIG. 52, the controller (1000) positions the droplet suction device (4000) in the second well. The controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that both the first syringe pump (3100) and the second syringe pump (3200) are connected to the hydrophobic fluid storage unit (8200), and controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200).

[0305] As can be seen in FIGS. 53 and 54, the controller (1000) performs a first droplet transfer operation and a second droplet transfer operation to transfer droplets of the second well through a common transfer channel. Since the operation of the controller (1000) in the first droplet transfer operation and the second droplet transfer operation is the same as in FIGS. 44 and 45, respectively, a detailed description will be omitted.

[0306] [5] Example 5

[0307] FIG. 55 shows the configuration of a droplet analysis device (10) according to Example 5. The droplet analysis device (10) according to Example 5 includes a controller (1000), a droplet suction device (4000), a droplet storage unit (9000), a hydrophilic fluid storage unit (8100), a hydrophobic fluid storage unit (8200), a droplet detection channel (6000), a valve (5500), a T-junction member (5700), a droplet processing unit (7000), a multi-port valve (5600), a first syringe pump (3100), and a second syringe pump (3200). Additionally, the dotted lines in FIG. 55 represent channels to which the components of the droplet analysis device (10) are fluidly connected.

[0308] The controller (1000) can control the operation of the first syringe pump (3100), the second syringe pump (3200), the valve (5500), the multi-port valve (5600), and the droplet suction device (4000).

[0309] The droplet detection channel (6000) may have a central inlet, at least two side inlets, and an outlet. Additionally, the droplet detection channel (6000) may have an X junction and may include a central inlet channel extending between the central inlet and the X junction, at least two side channels extending between each of the at least two side inlets and the X junction, and an outlet channel extending between the outlet and the X junction.

[0310] Additionally, the T junction member (5700) is a fitting having a T junction and may have a junction inlet and at least two junction side outlets. Here, each of the at least two junction side outlets may be fluidly connected to each of the at least two side inlets of the droplet detection channel (6000).

[0311] The multi-port valve (5600) selectively connects the central inlet of the droplet detection channel (6000) to the droplet suction device (4000) or the first syringe pump (3100). Additionally, when the multi-port valve (5600) connects the central inlet of the droplet detection channel (6000) to the droplet suction device (4000), the multi-port valve (5600) fluidically connects the central inlet of the droplet detection channel (6000) to the internal channel of the droplet suction device (4000), while fluidly connecting the first syringe pump (3100) to the external channel of the droplet suction device (4000).

[0312] The first syringe pump (3100) can be connected to a hydrophobic fluid storage unit (8200). Additionally, the second syringe pump (3200) can be optionally connected to either the hydrophobic fluid storage unit (8200) or the hydrophilic fluid storage unit (8100). Additionally, the second syringe pump (3200) is fluidically connected to at least two side inlets of the droplet detection channel (6000) via a T-junction member (5700).

[0313] The descriptions of the other components are the same as those above, so they are omitted.

[0314] Additionally, the droplet analysis device (10) according to Example 5 may include a configuration not disclosed in FIG. 28. For example, at least one fluorescence detector (2000) may be arranged on one side of the outlet channel of the droplet detection channel (6000). At this time, since the operation and function of the at least one fluorescence detector (2000) are the same as described above, a description will be omitted.

[0315]

[0316] FIGS. 56 to 58 are for explaining the droplet transfer operation of the droplet analysis device (10) according to Example 5. Referring to FIG. 56, the controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that both the first syringe pump (3100) and the second syringe pump (3200) are connected to the hydrophobic fluid storage unit (8200), and controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200). And the controller (1000) controls the syringe pump (3100) so that the syringe pump (3100) provides hydraulic pressure so that the loaded hydrophobic fluid fills all the channels indicated by the dotted line in FIG. 55.

[0317] Referring to FIG. 57, the controller (1000) performs a first droplet transfer operation. For example, the controller (1000) closes (OFF) the valve (5500) and positions the droplet suction device (4000) in the first well of the droplet storage unit (9000). Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the central inlet of the droplet detection channel (6000) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0318] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that hydrophobic fluid is transferred to the external channel of the droplet suction device (4000). The controller (1000) controls the second syringe pump (3200) to apply negative pressure so that droplets of the first well are positioned between the multi-port valve (5600) and the central inlet of the droplet detection channel (6000).

[0319] Referring to FIG. 58, after performing the first droplet transfer operation, the controller (1000) performs the second droplet transfer operation. For example, the controller (1000) opens the valve (5500) and controls the multi-port valve (5600) so that the first syringe pump (3100) and the central inlet of the droplet detection channel (6000) are fluidly connected. At this time, the controller (1000) can move the droplet suction device (4000) to a designated position. At this time, the designated position is a position where the droplet or fluid emitted from the internal channel of the droplet suction device (4000) is transferred to the droplet processing unit (7000). However, the controller (1000) does not necessarily move the droplet suction device (4000) to a designated position during the second droplet transfer operation, and the controller (1000) may move the droplet suction device (4000) to a designated position before the second droplet transfer operation and the hydrophilic fluid cleaning of FIGS. 60 to 61 described later.

[0320] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that droplets located between the multi-port valve (5600) and the central inlet of the droplet detection channel (6000) pass through the droplet detection channel (6000). At this time, the controller (1000) controls the second syringe pump (3200) to apply positive pressure so that droplets moving from the central inlet channel to the outlet channel of the droplet detection channel (6000) can meet the hydrophobic fluid injected into at least two side channels of the droplet detection channel (6000) at the X-junction. Droplets that meet the hydrophobic fluid at the X-junction are sufficiently separated from each other.

[0321] While droplets pass through the outlet channel of the droplet detection channel (6000), the controller (1000) controls at least one fluorescence detector to detect fluorescence from the droplets. The droplets that have passed through the droplet detection channel (6000) are transferred to the droplet processing unit (7000) via the valve (5500).

[0322] Referring to the first droplet transfer operation and the second droplet transfer operation, the droplets of the first well pass through the internal channel of the droplet suction device (4000), the multi-port valve (5600), and the central inlet channel and outlet channel of the droplet detection channel (6000). The path along which the droplets of the first well have traveled is called the common movement channel, and some of the droplets of the first well remain in the common movement channel.

[0323] Referring to FIG. 59, after the first and second droplet transfer operations for the first well are performed, the controller (1000) controls the second syringe pump (3100) so that the second syringe pump (3100) and the hydrophilic fluid storage unit (8100) are fluidly connected. Additionally, the controller (1000) controls the first syringe pump (3100) so that the first syringe pump (3100) loads hydrophobic fluid from the hydrophobic fluid storage unit (8200), and controls the second syringe pump (3200) so that the second syringe pump (3200) loads hydrophilic fluid from the hydrophilic fluid storage unit (8100).

[0324] Referring to FIGS. 60 to 61, the controller (1000) can perform a hydrophilic fluid cleaning operation. For example, the controller (1000) can close (OFF) the valve (5500). For example, the controller (1000) may keep the valve (5500) closed while performing the hydrophilic fluid cleaning operation. Alternatively, the controller (1000) may close the valve (5500) only during a portion of the time interval in which the hydrophilic fluid cleaning operation is performed, and open it during the remaining interval. For example, the controller (1000) may close the valve (5500) while starting the hydrophilic fluid cleaning operation and open the valve (5500) after a first time has elapsed, or it may open the valve (5500) when starting the hydrophilic fluid cleaning operation and close the valve (5500) after a second time has elapsed. Here, the first time and the second time may be the same or different. At this time, the droplet suction device (4000) is positioned at the designated location. Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the central inlet of the droplet detection channel (6000) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0325] The controller (1000) can control the first syringe pump (3100) to apply positive pressure so that hydrophobic fluids are transferred to the external channel of the droplet suction device (4000), but this may be omitted. During the time interval when the valve (5500) is closed, as can be seen in FIG. 60, the controller (1000) can control the second syringe pump (3200) to apply positive pressure so that hydrophilic fluids can be transferred to the droplet processing unit (7000) through the X-junction of the droplet detection channel (6000), the multi-port valve (5600), and the internal channel of the droplet suction device (4000). Additionally, during the time interval when the valve (5500) is open, as can be seen in FIG. 61, the controller (1000) described above controls the second syringe pump (3200) to apply positive pressure, so that a hydrophilic fluid flows into at least two junction side inlets of the X-junction member (5400), passes through the X-junction member (5400), passes through the droplet suction device (4000), and also passes through the droplet detection channel (6000).

[0326] Accordingly, residual droplets in a portion of the common moving channel leading to the X-junction of the droplet detection channel (6000), the multi-port valve (5600), and the internal channel of the droplet suction device (4000) are washed by a hydrophilic fluid.

[0327] After the hydrophilic fluid washing operation, the controller (1000) controls the second syringe pump (3200) so that the second syringe pump (3200) and the hydrophobic fluid storage unit (8200) are connected as shown in FIG. 62.

[0328] Additionally, the controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200).

[0329] Referring to FIG. 63, the controller (1000) performs a first hydrophobic fluid cleaning operation. For example, the controller (1000) opens the valve (5500) and controls the multi-port valve (5600) so that the first syringe pump (3100) and the central inlet of the droplet detection channel (6000) are fluidly connected. The controller (1000) controls the first syringe pump (3100) so that the first syringe pump (3100) applies positive pressure to the central inlet of the droplet detection channel (6000). The application of positive pressure by the first syringe pump (3100) causes the hydrophilic fluid, which was filled between the multi-port valve (5600) and the central inlet of the droplet detection channel (6000) during the hydrophilic fluid washing operation, to move to the droplet processing unit (7000) by passing through the central inlet channel and outlet channel of the droplet detection channel (6000). Accordingly, the remaining droplets in the droplet detection channel (6000) that were not washed during the hydrophilic fluid washing operation are washed by the hydrophilic fluid.

[0330] Additionally, after the remaining droplets are washed by a hydrophilic fluid, the channel connecting the first syringe pump (3100), the multi-port valve (5600), the X-junction member (5400), and the droplet detection channel (6000) is washed once more by a hydrophobic fluid. Meanwhile, the controller (1000) controls the second syringe pump (3200) so that the second syringe pump (3200) applies positive pressure to at least two side inlets of the droplet detection channel (6000). Accordingly, the channel leading to the second syringe pump (3200) and the droplet detection channel (6000) is washed by a hydrophobic fluid.

[0331] Referring to FIG. 64, the controller (1000) performs a second hydrophobic fluid cleaning operation. For example, the controller (1000) can close the valve (5500) (OFF). However, as mentioned in the description of the hydrophilic fluid cleaning operation above, the controller (1000) may close the valve (5500) only during a portion of the time interval in which the second hydrophobic fluid cleaning operation is performed, and open it during the remaining interval. At this time, the droplet suction device (4000) is located at the designated position. Additionally, the controller (1000) controls the multi-port valve (5600) so that the internal channel of the droplet suction device (4000) and the central inlet of the droplet detection channel (6000) are fluidly connected, and the external channel of the first syringe pump (3100) and the droplet suction device (4000) are fluidly connected.

[0332] The controller (1000) controls the first syringe pump (3100) to apply positive pressure so that hydrophobic fluids are transferred to the external channel of the droplet suction device (4000). The controller (1000) controls the second syringe pump (3200) to apply positive pressure so that hydrophobic fluids can be transferred to the droplet processing unit (7000) through the droplet detection channel (6000), the multi-port valve (5600), and the internal channel of the droplet suction device (4000).

[0333] When the hydrophobic fluid washing operation is completed, the controller (1000) performs a droplet transfer operation to measure droplets in the second well that are different from the first well. Referring to FIG. 65, the controller (1000) positions the droplet suction device (4000) in the second well. The controller (1000) controls the first syringe pump (3100) and the second syringe pump (3200) so that both the first syringe pump (3100) and the second syringe pump (3200) are connected to the hydrophobic fluid storage unit (8200), and controls the first syringe pump (3100) and the second syringe pump (3200) so that the first syringe pump (3100) and the second syringe pump (3200) load hydrophobic fluid from the hydrophobic fluid storage unit (8200).

[0334] As can be seen in FIGS. 66 and 67, the controller (1000) performs a first droplet transfer operation and a second droplet transfer operation to transfer droplets of the second well through a common transfer channel. Since the operation of the controller (1000) in the first droplet transfer operation and the second droplet transfer operation is the same as that in FIGS. 57 and 58, respectively, a detailed description will be omitted.

[0335]

[0336] [6] Example 6

[0337] FIG. 68 shows the configuration of a droplet analysis device (10) according to Example 6. The droplet analysis device (10) according to Example 6 has the same configuration as the droplet analysis device (10) according to Example 5. In addition, the controller (1000) of the droplet analysis device (10) according to Example 6 operates in the same way as the controller of the droplet analysis device according to Example 5.

[0338] Referring to FIGS. 69 to 71, the controller (1000) of the droplet analysis device (1000) according to Embodiment 6 performs first and second droplet transfer operations for the droplets of the first well. At this time, the operation of the controller (1000) is the same as the operation described in FIGS. 56 to 58 in Embodiment 5.

[0339] Referring to FIGS. 72 to 74, the controller (1000) of the droplet analysis device (1000) according to Embodiment 6 performs a hydrophilic fluid washing operation. At this time, the operation of the controller (1000) is the same as the operation described in FIGS. 59 to 61 in Embodiment 5.

[0340] Referring to FIGS. 75 to 77, the controller (1000) of the droplet analysis device (1000) according to Embodiment 6 performs first and second hydrophobic fluid washing operations. At this time, the operation of the controller (1000) is the same as the operation described in FIGS. 62 to 64 in Embodiment 5.

[0341] Referring to FIGS. 78 to 80, the controller (1000) of the droplet analysis device (1000) according to Embodiment 6 performs first and second droplet transfer operations for the droplets of the second well. At this time, the operation of the controller (1000) is the same as the operation described in FIGS. 65 to 67 in Embodiment 5.

[0342] However, the structure of the droplet detection channel (6000) in Example 6 is different from that of Example 5. Referring to FIGS. 68 through 80, the droplet detection channel (6000) in Example 6 may have a central inlet, at least two side inlets, and an outlet, just like the droplet detection channel in Example 5. Additionally, the droplet detection channel (6000) may have an X junction and include a central inlet channel extended between the central inlet and the X junction, at least two side channels extended between each of the at least two side inlets and the X junction, and an outlet channel extended between the outlet and the X junction. However, the droplet detection channel (6000) in Example 6 has at least two T junctions (6200) disposed in each of the at least two side channels. Additionally, the droplet detection channel (6000) has at least one cross-junction (6300) disposed in the outlet channel. Additionally, the droplet detection channel (6000) has at least one branch channel extending between one of at least two T-junctions (6200) and one of at least one cross-junction (6300).

[0343] Referring to FIGS. 70, 77 and 79, in the first droplet transfer operation and the second hydrophobic fluid washing operation, when the controller (1000) controls the second syringe pump (3200) to apply negative pressure to at least two side junctions of the droplet detection channel (6000), the hydrophobic fluid moves from the droplet detection channel (6000) to at least one cross junction and to at least two T junctions.

[0344] Referring to FIG. 73, in a hydrophilic fluid cleaning operation, when the controller (1000) controls the second syringe pump (3200) to apply negative pressure to at least two side junctions of the droplet detection channel (6000), the hydrophilic fluid moves from the droplet detection channel (6000) to at least one cross junction and to at least two T junctions. Accordingly, the remaining droplets in the outlet channel up to the cross junction closest to the outlet of the droplet detection channel (6000) are also cleaned in the hydrophilic fluid cleaning operation, so the common movement channel can be cleaned more than in FIG. 60 of Embodiment 5. In other words, in FIG. 60 of Embodiment 5, during the hydrophilic fluid cleaning operation, the hydrophilic fluid reaches only up to the X junction of the droplet detection channel (6000), so the outlet channel of the droplet detection channel (6000) is not cleaned in the hydrophilic fluid cleaning operation.

[0345] However, in FIG. 73 of Example 6, since the hydrophilic fluid reaches the cross junction, the outlet channel of the droplet detection channel (6000) is also cleaned in the hydrophilic fluid cleaning operation.

[0346]

[0347] [Experiment on cleaning effect when adding hydrophilic cleaning action]

[0348] [Preparation of High-Concentration Sample Droplets and NTC Droplets for Experiment]

[0349] Here, high-concentration sample droplets refer to droplets obtained by heat-treating and amplifying a droplet generated by mixing sample DNA, primers, and a premix. Additionally, NTC droplets are droplets obtained by heat-treating and amplifying a droplet generated by mixing primers and a premix without sample DNA.

[0350] To prepare a high-concentration sample droplet, the applicant mixed sample DNA containing a pre-prepared target DNA sequence, primers capable of amplifying the target DNA, and other required premixes. Here, the sample DNA is not actual DNA but artificial DNA created for the experiment. Accordingly, primers capable of binding to and amplifying the artificially created DNA were used. Here, the concentration of the sample DNA was 10 pmol / ul.

[0351] Subsequently, a sample containing a mixture of sample DNA, primers, and a premix was placed in a BioTNS cartridge, and the cartridge containing the sample was loaded into a BioTNS TGR-100. The TGR-100 was then operated to generate sample droplets. High-concentration sample droplets with amplified sample DNA were prepared by heat-treating the generated sample droplets using a BioRad T100 THERMAL CYCLER.

[0352] Meanwhile, to prepare the NTC sample droplet, the applicant mixed the same primers and premixes used to prepare the high-concentration sample droplet. At this time, the sample DNA was not mixed, and the amounts of primers and premixes used for mixing were identical to the amounts used to generate the high-concentration sample droplet.

[0353] Subsequently, sample droplets were generated using BioTNS’s TGR-100 as described above, and NTC sample droplets were prepared using RioRad’s T100 THERMAL CYCLER.

[0354]

[0355] *[Preparation of Well Plate]

[0356] A first well plate and a second well plate were prepared to perform comparative and experimental examples. High-concentration sample droplets were placed in two wells of each of the first well plate and the second well plate, and NTC sample droplets were placed in two wells.

[0357] [Comparison Example]

[0358] In the comparative example, the fluorescence intensity of high-concentration sample droplets and NTC sample droplets contained in a first-well plate was measured using BioTNS’s TAS-100. Specifically, the fluorescence intensity of high-concentration sample droplets contained in one of the two wells of the first-well plate containing high-concentration sample droplets was measured. Subsequently, a hydrophobic fluid wash operation was performed, and the fluorescence intensity of NTC sample droplets contained in one of the two wells of the first-well plate containing NTC sample droplets was measured. Subsequently, a hydrophobic fluid wash operation was performed, and the fluorescence intensity of high-concentration sample droplets contained in the remaining well of the two wells of the first-well plate containing high-concentration sample droplets was measured. Subsequently, a hydrophobic fluid wash operation was performed, and the fluorescence intensity of NTC sample droplets contained in the remaining well of the two wells of the first-well plate containing NTC sample droplets was measured.

[0359] [Experimental Example]

[0360] In the experimental example, the fluorescence intensity of high-concentration sample droplets and NTC sample droplets contained in the second well plate was measured using the droplet analysis device disclosed in FIG. 55. Specifically, the fluorescence intensity of high-concentration sample droplets contained in one of the two wells of the second well plate containing high-concentration sample droplets was measured. Subsequently, a hydrophilic fluid washing operation and a hydrophobic fluid washing operation were performed, and the fluorescence intensity of NTC sample droplets contained in one of the two wells of the second well plate containing NTC sample droplets was measured. Subsequently, a hydrophilic fluid washing operation and a hydrophobic fluid washing operation were performed, and the fluorescence intensity of high-concentration sample droplets contained in the remaining well of the two wells of the second well plate containing high-concentration sample droplets was measured. Subsequently, a hydrophilic fluid washing operation and a hydrophobic fluid washing operation were performed, and the fluorescence intensity of NTC sample droplets contained in the remaining well of the two wells of the second well plate containing NTC sample droplets was measured.

[0361] [Experimental Results]

[0362] FIGS. 81 and 82 show the results of measuring fluorescence intensity of a comparative example, and FIGS. 83 and 84 show the results of measuring fluorescence intensity of an experimental example. In other words, FIG. 81 is the result of measuring the fluorescence intensity of high-concentration sample droplets contained in one of the two wells containing high-concentration sample droplets of the first plate and NTC sample droplets contained in one of the two wells containing high-concentration sample droplets of the first plate, and FIG. 82 is the result of measuring the fluorescence intensity of high-concentration sample droplets contained in the remaining well of the two wells containing high-concentration sample droplets of the first plate and NTC sample droplets contained in the remaining well of the two wells containing high-concentration sample droplets of the first plate.

[0363] FIG. 83 is the result of measuring the fluorescence intensity of high-concentration sample droplets contained in one of the two wells containing high-concentration sample droplets of the second plate and NTC sample droplets contained in one of the two wells containing high-concentration sample droplets of the second plate, and FIG. 84 is the result of measuring the fluorescence intensity of high-concentration sample droplets contained in the remaining well of the two wells containing high-concentration sample droplets of the second plate and NTC sample droplets contained in the remaining well of the two wells containing high-concentration sample droplets of the second plate.

[0364] In FIGS. 81 to 84, one dot represents one fluorescence intensity value, the vertical axis represents the fluorescence intensity value, and the horizontal axis represents the number of droplets.

[0365] In the case of the comparison example, when the first NTC droplet is measured, it can be seen that significant noise, consisting of dots with fluorescence intensity values ​​that cannot be measured, is detected. Additionally, it can be seen that more noise is detected in the second NTC droplet measurement compared to the first NTC droplet measurement.

[0366] In the case of the experimental example, it can be seen that almost no noise was detected when the first NTC droplet was measured. In addition, although the noise increased in the second NTC droplet measurement compared to the first NTC droplet measurement, it can be seen that the noise was significantly reduced compared to when only the hydrophobic fluid washing operation was performed.

[0367] [Analysis of Experimental Results]

[0368] Referring to FIGS. 81 to 84, it can be seen that when a hydrophilic fluid washing operation is performed before a hydrophobic fluid washing operation, the remaining droplets in the droplet detection channel that detects the fluorescence of the droplets are significantly reduced.

[0369] As explained above, when a hydrophilic fluid is used, residual droplets adhering to the inner walls of channels due to surface tension or to stepped areas such as valves are dissolved and washed away, indicating that the cleaning performance of residual droplets is improved compared to when only a hydrophobic fluid cleaning operation is performed.

[0370]

[0371] Although the embodiments have been described above with reference to limited examples and drawings, those skilled in the art can make various modifications and variations from the description above. For example, suitable results can be achieved even if the described techniques are performed in a different order than described, and / or the components of the described system, structure, device, circuit, etc. are combined or assembled in a form different from described, or replaced or substituted by other components or equivalents.

[0372] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims set forth below.

Claims

1. In a droplet analysis device for analyzing droplets, A detection channel having a detection inlet and a detection outlet; A first valve connected to the above detection outlet; A droplet suction device for drawing in droplets to transfer droplets contained in a designated well to the detection channel; X junction component having a junction center inlet, at least two junction side inlets, and a junction outlet - wherein the junction outlet is connected to the detection inlet -; Hydrophobic fluid reservoir for storing hydrophobic fluid; A hydrophilic fluid reservoir for storing hydrophilic fluids; A first path control unit that selectively connects the above droplet suction unit and the above junction central inlet; A second path control unit that selectively connects the above hydrophilic fluid reservoir and the above at least two junction side inlets; A third path control unit that selectively connects the above hydrophobic fluid reservoir and the above at least two junction side inlets; A hydraulic supply unit that selectively supplies hydraulic pressure to the first to third path control units; and A controller that controls the operation of the first valve, the droplet suction device, the first path control unit, the second path control unit, the third path control unit, and the hydraulic pressure supply unit; comprising The above controller is, A hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and A first hydrophobic fluid washing operation is performed to allow the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; The above hydrophilic fluid washing operation is performed before the above first hydrophobic fluid washing operation, and The above controller is, The first valve is closed, the second path controller is controlled so that the hydrophilic fluid reservoir and the at least two junction side inlets are connected, the first path controller is controlled so that the droplet suction unit and the junction center inlet are connected, and the hydraulic supply unit is controlled to provide hydraulic pressure to the second path controller, thereby performing the hydrophilic fluid cleaning operation. The above controller is, Close the first valve, control the third path controller so that the hydrophobic fluid reservoir and the at least two junction side inlets are connected, control the first path controller so that the droplet suction unit and the junction center inlet are connected, and control the hydraulic supply unit to provide hydraulic pressure to the third path controller to perform the first hydrophobic fluid cleaning operation. Droplet analysis device.

2. In Paragraph 1, The above controller is, A second hydrophobic fluid washing operation is performed to allow the hydrophilic fluid filled between the X junction member and the second path control member to move to the detection channel in the above hydrophilic fluid washing operation; The above second hydrophobic fluid washing operation is performed between the above hydrophilic fluid washing operation and the above first hydrophobic fluid washing operation, and The above controller is, Opening the first valve, controlling the first path control unit so that the hydrophobic fluid reservoir and the junction central inlet are connected, and controlling the hydraulic supply unit to provide hydraulic pressure to the first path control unit to perform the second hydrophobic fluid cleaning operation. Droplet analysis device.

3. In Paragraph 1, The above controller is, A first droplet transfer operation for transferring droplets contained in the designated well between the first path control unit and the X junction member; and A second droplet transfer operation is performed to transfer droplets between the first path control unit and the X junction member to the detection channel; and The above controller is, The above droplet suction device is moved to the above-designated well, the first valve is closed, the third path control unit is controlled so that the at least two junction side inlets are connected, the first path control unit is controlled so that the droplet suction device and the junction center inlet are connected, and the hydraulic supply unit is controlled to provide hydraulic pressure to the third path control unit, thereby performing the first droplet transfer operation. Opening the first valve, controlling the first path control unit so that the droplet suction unit and the junction center inlet are not connected, and controlling the hydraulic supply unit to provide hydraulic pressure to the first path control unit to perform the second droplet transfer operation. Droplet analysis device.

4. In Paragraph 3, The above controller is, While performing the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, the first droplet transfer operation and the second droplet transfer operation are also performed after the hydrophobic fluid washing operation, Prior to the above hydrophilic fluid washing operation, The controller moves the droplet suction device to the first well in the first droplet transfer operation, transfers the droplets contained in the first well between the first path control unit and the X junction member, and After the above hydrophobic fluid washing operation, The controller moves the droplet suction device to a second well different from the first well in the first droplet transfer operation, thereby transferring the droplets contained in the second well between the first path control unit and the X junction member. Droplet analysis device.

5. In Paragraph 1, The above controller is, Control the above droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, The first position is a position where a droplet storage unit containing the designated well is not placed, Droplet analysis device.

6. In a droplet analysis device for analyzing droplets, A detection channel having a detection inlet and a detection outlet; A first valve connected to the above detection outlet; A droplet suction device for drawing in droplets to transfer droplets contained in a designated well to the detection channel; X junction component having a junction center inlet, at least two junction side inlets, and a junction outlet - wherein the junction outlet is connected to the detection inlet -; Hydrophobic fluid reservoir for storing hydrophobic fluid; A hydrophilic fluid reservoir for storing hydrophilic fluids; A first pump that provides hydraulic pressure to the central inlet of the above-mentioned junction; A second pump that provides hydraulic pressure to the above at least two junction side inlets; A second valve that allows the central inlet of the junction to be optionally connected to the droplet suction device or the first pump; and A controller controlling the first valve, the second valve, the droplet suction device, the first pump, and the second pump; comprising The above controller is, A hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the X junction member and the droplet suction device; and A first hydrophobic fluid washing operation is performed to allow the hydrophobic fluid to move from the hydrophobic fluid reservoir to the X junction member and the droplet suction device; The above hydrophilic fluid washing operation is performed before the above first hydrophobic fluid washing operation, and The above controller is, Close the first valve, control the second pump so that the hydrophilic fluid reservoir and the second pump are connected, control the second valve so that the droplet suction device and the junction center inlet are connected, and control the second pump so that the second pump provides hydraulic pressure to the at least two side inlets to perform the hydrophilic fluid cleaning operation. The above controller is, Close the first valve, control the second pump so that the hydrophobic fluid reservoir and the second pump are connected, control the second valve so that the droplet suction device and the junction center inlet are connected, and control the second pump so that the second pump provides hydraulic pressure to the at least two side inlets to perform the first hydrophobic fluid cleaning operation. Droplet analysis device.

7. In Paragraph 6, The above controller is, A second hydrophobic fluid washing operation is performed to allow the hydrophilic fluid filled between the X junction member and the second valve in the above hydrophilic fluid washing operation to move to the detection channel; The above second hydrophobic fluid washing operation is performed between the above hydrophilic fluid washing operation and the above first hydrophobic fluid washing operation, and The above controller is, Opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the junction center inlet and the first pump are connected, and controlling the first pump to provide hydraulic pressure to the junction center inlet to perform the second hydrophobic fluid cleaning operation. Droplet analysis device.

8. In Paragraph 6, The above controller is, A first droplet transfer operation for transferring droplets contained in the designated well between the second valve and the X junction member; and A second droplet transfer operation is performed to transfer droplets between the second valve and the X junction member to the detection channel; and The above controller is, The droplet suction device is moved to the designated well, the first valve is closed, the second pump is controlled so that the hydrophobic fluid reservoir and the second pump are connected, the second valve is controlled so that the droplet suction device and the junction center inlet are connected, and the second pump is controlled so that the second pump provides hydraulic pressure to the at least two junction side inlets, thereby performing the first droplet transfer operation. The above controller is, Opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the first pump and the junction center inlet are connected, and controlling the first pump so that the first pump provides hydraulic pressure to the junction center inlet, thereby performing the second droplet transfer operation. Droplet analysis device.

9. In Paragraph 8, The above controller is, While performing the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, the first droplet transfer operation and the second droplet transfer operation are also performed after the hydrophobic fluid washing operation, Prior to the above hydrophilic fluid washing operation, The controller moves the droplet suction device to the first well in the first droplet transfer operation, transfers the droplets contained in the first well between the second valve and the X junction member, and After the above hydrophobic fluid washing operation, The controller moves the droplet suction device to a second well different from the first well in the first droplet transfer operation, thereby transferring the droplets contained in the second well between the second valve and the X junction member. Droplet analysis device.

10. In Paragraph 6, The above controller is, Control the above droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, The first position is a position where a droplet storage unit containing the designated well is not placed, Droplet analysis device.

11. In a droplet analysis device for analyzing droplets, A plate having a central inlet, at least two side inlets and an outlet - wherein the plate has an X junction, and the plate comprises (i) a central channel extending between the central inlet and the X junction, (ii) at least two side channels extending between each of the at least two side inlets and the X junction, and (iii) an outlet channel extending between the outlet and the X junction - ; A first valve connected to the above outlet; A droplet suction device for drawing in droplets to transfer droplets contained in a designated well to the plate; Hydrophobic fluid reservoir for storing hydrophobic fluid; A hydrophilic fluid reservoir for storing hydrophilic fluids; A first pump that provides hydraulic pressure to the central inlet above; A second pump that provides hydraulic pressure to the above at least two side inlets; A second valve that allows the central inlet to be optionally connected to the droplet suction device or the first pump; and A controller controlling the first valve, the second valve, the droplet suction device, the first pump, and the second pump; comprising The above controller is, A hydrophilic fluid washing operation that allows the hydrophilic fluid to move from the hydrophilic fluid reservoir to the plate and the droplet suction device; and A first hydrophobic fluid washing operation that allows the hydrophobic fluid to move from the hydrophobic fluid reservoir to the plate and the droplet suction device; wherein The above hydrophilic fluid washing operation is performed before the above first hydrophobic fluid washing operation, and The above controller is, Close the first valve, control the second pump so that the hydrophilic fluid reservoir and the second pump are connected, control the second valve so that the droplet suction device and the central inlet are connected, and control the second pump so that the second pump provides hydraulic pressure to the at least two side inlets to perform the hydrophilic fluid cleaning operation. The above controller is, Close the first valve, control the second pump so that the hydrophobic fluid reservoir and the second pump are connected, control the second valve so that the droplet suction device and the central inlet are connected, and control the second pump so that the second pump provides hydraulic pressure to the at least two side inlets to perform the hydrophobic fluid cleaning operation. Droplet analysis device.

12. In Paragraph 11, The above controller is, A second hydrophobic fluid washing operation is performed to allow the hydrophilic fluid filled between the X junction and the second valve to move to the plate in the above hydrophilic fluid washing operation; The above second hydrophobic fluid washing operation is performed between the above hydrophilic fluid washing operation and the above first hydrophobic fluid washing operation, and The above controller is, Opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the central inlet and the first pump are connected, and controlling the first pump to provide hydraulic pressure to the central inlet to perform the second hydrophobic fluid cleaning operation. Droplet analysis device.

13. In Paragraph 11, The above controller is, A first droplet transfer operation for transferring droplets contained in the designated well between the second valve and the plate; and A second droplet transfer operation is performed to transfer droplets between the second valve and the plate to the plate; and The above controller is, The droplet suction device is moved to the designated well, the first valve is closed, the second pump is controlled so that the hydrophobic fluid reservoir and the second pump are connected, the second valve is controlled so that the droplet suction device and the central inlet are connected, and the second pump is controlled so that the second pump provides hydraulic pressure to the at least two side inlets, thereby performing the first droplet transfer operation. Opening the first valve, controlling the first pump so that the hydrophobic fluid reservoir and the first pump are connected, controlling the second valve so that the first pump and the central inlet are connected, and controlling the first pump so that the first pump provides hydraulic pressure to the central inlet to perform the second droplet transfer operation. Droplet analysis device.

14. In Paragraph 13, The above controller is, While performing the first droplet transfer operation and the second droplet transfer operation before the hydrophilic fluid washing operation, the first droplet transfer operation and the second droplet transfer operation are also performed after the hydrophobic fluid washing operation, Prior to the above hydrophilic fluid washing operation, The controller moves the droplet suction device to the first well in the first droplet transfer operation, transfers the droplets contained in the first well between the second valve and the plate, and After the above hydrophobic fluid washing operation, The controller moves the droplet suction device to a second well different from the first well in the first droplet transfer operation, thereby transferring the droplets contained in the second well between the second valve and the plate. Droplet analysis device.

15. In Paragraph 11, The above controller is, Control the above droplet suction device to be positioned at a first position to perform the hydrophilic fluid washing operation and the hydrophobic fluid washing operation, The first position is a position where a droplet storage unit containing the designated well is not placed, Droplet analysis device.

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