Liquid Droplet Quality Control Method, Device, Storage Medium and Program Product Based on Laser Detection

Abstract

The present invention discloses a droplet quality control method, device, storage medium and program product based on laser detection. The method includes: acquiring droplet microchannel signal data collected by a laser device, and monitoring the state of droplets in the droplet microchannel based on the droplet microchannel signal data; wherein the droplet microchannel includes each single microchannel and / or a main microchannel, the droplets in the droplet microchannel include the droplets in each single microchannel and / or the fused droplets in the main microchannel, the droplet microchannel signal data includes single microchannel signal data and / or main microchannel signal data. After the solutions corresponding to the respective single microchannels form the droplets in the respective single microchannels, the droplets in the respective single microchannels flow into the main microchannel and fuse in the main microchannel to form fused droplets.

Description

Technical Field

[0001] The present invention relates to the field of gene technology, and particularly to a droplet quality control method, device, storage medium and program product based on laser detection. Background Art

[0002] Droplet microfluidics technology, as an important bioanalysis tool, has been widely applied in fields such as single-cell analysis, digital polymerase chain reaction (PCR), and immunoassay. By encapsulating single cells or reaction components in tiny oil droplets, droplet technology provides unprecedented possibilities for high-throughput parallel processing. However, with the in-depth development of life science research and the increasing application requirements, the simple droplet encapsulation technology has gradually shown its limitations and is difficult to meet the increasingly complex experimental needs. The main manifestations are as follows: (1) Increasing demand for complex reactions: The development of modern biotechnology not only requires encapsulating samples in droplets for simple independent reactions but also more complex operations such as fusing, separating, exchanging contents between droplets, and multi-step reaction chains. These operations require precise manipulation inside the droplets to achieve more complex and delicate experimental designs. (2) Balancing high throughput and diversity: In large-scale bioanalysis and drug screening, not only high throughput is required, but also content exchange and mixing between multiple droplets are required to simulate complex biological reaction environments. This demand poses higher requirements on droplet technology, which needs to process and fuse a large number of droplets with different components in a short time and ensure the consistency and reliability of each operation. (3) Multi-omics joint analysis: In single-cell multi-omics analysis, such as the joint analysis of transcriptome, proteome, and metabolome, it is necessary to separately extract, react, and analyze different contents from the same cell source. This demand requires droplet technology to flexibly achieve the separation and recombination of contents to ensure the integrity and relevance of data.

[0003] Therefore, the droplet fusion technology in the prior art faces many challenges: Unstable fusion success rate: The instability during the droplet fusion process remains a major problem. If the fusion efficiency is low or the fusion is incomplete, it will lead to sample loss and data deviation, affecting the reliability of experimental results. Lack of real-time monitoring means: Currently, the success or failure of droplet fusion often depends on post hoc analysis, lacking effective real-time monitoring means to ensure the success of the fusion process. In this case, errors in the experiment are difficult to be discovered and corrected in time, increasing the failure rate of the experiment. Complicated operating conditions: To achieve efficient droplet fusion, precise requirements are imposed on operating conditions such as droplet generation frequency, fusion electric field intensity, droplet position, and size. The control and optimization of these conditions are a major obstacle to the commercialization of droplet fusion technology. Summary of the Invention

[0004] To solve the existing technical problems, the embodiments of the present invention provide a droplet quality control method based on laser detection, a droplet quality control device based on laser detection, a single-cell sequencing library construction device, a computer-readable storage medium, and a computer program product, which can monitor the abnormal states in the whole process of droplet generation, pairing, and fusion in real time, facilitating maintenance personnel to detect and eliminate abnormalities in a timely manner.

[0005] In a first aspect, a droplet quality control method based on laser detection is provided, including: obtaining droplet microchannel signal data collected by a laser device, and monitoring the state of droplets in the droplet microchannel based on the droplet microchannel signal data; wherein the droplet microchannel includes each single microchannel and / or the main microchannel, the droplets in the droplet microchannel include the droplets in each single microchannel and / or the fused droplets in the main microchannel, the droplet microchannel signal data includes single microchannel signal data and / or main microchannel signal data, after the solutions corresponding to each single microchannel form the droplets in their respective single microchannels, the droplets in each single microchannel flow into the main microchannel and fuse in the main microchannel to form fused droplets.

[0006] In a second aspect, a droplet quality control device based on laser detection is provided, including a memory and a controller. When the computer program stored in the memory is executed by the controller, the controller is caused to execute the steps of the droplet quality control method provided by the embodiments of the present application.

[0007] In a third aspect, a single-cell sequencing library construction device is provided, including the droplet quality control device based on laser detection provided by the embodiments of the present application.

[0008] In a fourth aspect, a computer-readable storage medium is provided, on which a computer program is stored, and the computer program is executed by a controller to perform the steps of the droplet quality control method based on laser detection.

[0009] In a fifth aspect, a computer program product includes a computer program, and when the computer program is executed by a controller, it implements the steps of the droplet quality control method based on laser detection provided in the first aspect.

[0010] In the embodiments of the present application, the laser device is used to collect the real-time droplet microchannel signal data during the droplet monitoring process. The droplet microchannel signal data includes at least one of the following: single microchannel signal data, main microchannel signal data. By analyzing the single microchannel signal data and / or the main microchannel signal data, the droplet conditions in each single microchannel and / or the conditions of the fused droplets generated in the main microchannel can be monitored, so that the abnormal states in the whole process of droplet generation, pairing, and fusion can be monitored in real time, facilitating maintenance personnel to detect and eliminate abnormalities in a timely manner. Description of the Drawings

[0011] Figure 1 Schematic block diagram of a droplet quality control device based on laser detection in an embodiment;

[0012] Figure 2 Schematic diagram of a microchannel for generating droplets in a droplet quality control device based on laser detection in an embodiment;

[0013] Figure 3 Schematic diagram of droplet fusion in a droplet quality control device based on laser detection in an embodiment;

[0014] Figure 4 Flowchart of a droplet quality control method based on laser detection in an embodiment;

[0015] Figure 5 Schematic diagram of signal data of a single microchannel in an embodiment;

[0016] Figure 6 Schematic diagram of microchannel signal data corresponding to a standard object in an embodiment;

[0017] Figure 7 Schematic diagram of the pulse signal of a fused droplet with a target in an embodiment;

[0018] Figure 8 Schematic diagram of a droplet quality control device based on laser detection in an embodiment;

[0019] Figure 9 Schematic structural diagram of a droplet quality control device based on laser detection in an embodiment. Detailed implementation manners

[0020] The technical solution of the present invention will be further elaborated in detail below in conjunction with the accompanying drawings of the specification and specific embodiments.

[0021] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this invention belongs. The terms used in the specification of this invention herein are only for the purpose of describing specific embodiments and are not intended to limit the protection scope of this invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0022] In the following description, the expression "some embodiments" describes a subset of all possible embodiments. However, it should be understood that "some embodiments" can be the same subset or different subsets of all possible embodiments and can be combined with each other without conflict.

[0023] Microfluidics refers to a technology for manipulating fluids in a micron-scale space. This technology can miniaturize the basic functions of chemical, biological, and other laboratories onto a chip of a few square centimeters, so it is also known as Lab-on-a-chip. The microfluidic chip system can manipulate the flow of fluids in tiny channels or components with dimensions ranging from dozens to hundreds of microns, and the volume of the fluid being manipulated can be as small as 10 -9 ~10 -18 L. It integrates the basic operations of biochemical experiments such as sample reaction, preparation, separation, and detection onto a very small chip. A network of microchannels formed by controllable fluids runs through the microfluidic system, realizing the various functions of a conventional biochemical laboratory while reducing the cost of analysis and detection, accelerating the reaction speed, improving the reaction efficiency, and making the experiment more controllable.

[0024] Generating droplets on a microfluidic chip is a process in which one-phase fluid is dispersed in another immiscible or partially immiscible fluid. For two immiscible liquids, one is used as the continuous phase and the other as the dispersed phase. The dispersed phase is dispersed in the continuous phase in the form of tiny volume units (10 -15 ~10 -9 L) to form droplets. Currently, the methods for forming droplets can be divided into two categories: passive methods and active methods. Passive methods refer to controlling the generation of droplets by controlling the microchannel structure and the flow rate ratio of the two phases, while active methods generally drive and control the generation of droplets by applying an external force.

[0025] After the droplets are generated, the cells or cell nuclei in each droplet release their nucleic acids (DNA or RNA) to prepare for amplification. Specific amplification techniques, such as multiple displacement amplification (MDA) or PCR amplification through droplets, are used to amplify the nucleic acids in the droplets.

[0026] See Figure 1, is a schematic block diagram of a droplet quality control device based on laser detection in an embodiment. The droplet quality control device based on laser detection includes, but is not limited to, the following devices or components: a controller 10, a gas generation device 11, an adjustment device 12, a laser device 13, a pressure detection device 14, a gas purification device 15, a pressure output device 16, and a display device 17. Among them, the controller 10 is respectively connected to each of the other devices. As the core of the droplet quality control device based on laser detection, the controller is responsible for coordinating and managing the functions of each component. The controller receives data from each device, processes and analyzes the data of each device, and generates different commands to control each device. The gas generation device 11 is used to provide a stable and adjustable air pressure for the entire experimental process. The gas generation device 11 is a key device for maintaining the required pressure environment for the experiment, ensuring that the experimental conditions meet the preset standards. The gas purification device 15 is used to remove moisture in the gas to prevent water vapor from affecting the experimental accuracy and equipment performance. Dry gas helps to maintain the stability of the system and extend the service life of the equipment. The air pressure generated by the gas generation device 11 is output to each single microchannel through the pressure output device 16 to maintain the pressure in each single microchannel. The pressure detection device 14 is used to detect the pressure level of each single microchannel in real time. Through the pressure detection device 14, the minute changes in the pressure in each single microchannel can be accurately measured and recorded, providing key data for the experiment. The adjustment device 12 is used to control the rate and amount of gas flowing into each single microchannel to ensure the uniformity and stability of the experimental conditions. The adjustment of the adjustment device 12 can be automatically adjusted according to the pressure data fed back by the pressure detection device 14 to respond to the changes in requirements during the experimental process. Among them, the adjustment device 12 is connected to the gas generation device 11. The laser device 13 is used to project a laser signal onto the detection area of the droplet microchannel. The laser signal encounters the droplet microchannel to form a detection signal, thereby obtaining droplet microchannel signal data, where the detection signal includes, but is not limited to, the signal formed by the reflection and / or refraction of the laser signal when it encounters the droplet microchannel. Through the laser device 13, the detection signal during the droplet monitoring process can be captured, facilitating the analysis of whether there are faults during the droplet monitoring process or whether the generated droplets are qualified based on the signals during the droplet monitoring process. The display device 17 can be a display component integrated on the droplet quality control device based on laser detection, or an external display device connected to the droplet quality control device based on laser detection, etc. The display device 17 is used to display the display data sent by the controller 10 for the user to view intuitively.

[0027] In an alternative implementation, for each individual microchannel, at least one laser device 13 can be configured to collect microchannel signal data corresponding to each individual microchannel. For the main microchannel, at least one laser device 13 can also be provided to collect main microchannel signal data corresponding to the main microchannel. It can be understood that the laser device 13 for the individual microchannel and the laser device 13 for the main microchannel can be configured simultaneously, or only one of them can be configured separately. Each laser device 13 corresponds to a laser detection area. When a droplet passes through the laser detection area, the signal data in the droplet microchannel at this time can be collected.

[0028] In some embodiments, the droplet quality control device based on laser detection may further include an image acquisition device. The image acquisition device is used to collect real-time images of the droplet microchannel during the droplet monitoring process and transmit the captured images and / or videos to the display device 17 in real time. Through the image acquisition device, detailed images of each instant during the droplet monitoring process can be captured, facilitating the analysis of whether there are faults during the droplet monitoring process or whether the generated droplets are qualified based on the images during the droplet monitoring process. The image acquisition device may include, but is not limited to, the following components: a high-frame-rate camera, a collimating lens, a fill light, etc. Among them, the high-frame-rate camera has high definition and high-speed image capture capabilities to capture rapidly changing experimental situations. The collimating lens is used to focus and direct light to precisely irradiate the photosensitive element to ensure image quality. The fill light is used to provide illumination in an environment with weak light so that clear images can be obtained even in a weak environment. The image acquisition device can be one or more. The adjustment device 12 can be one or more proportional valves.

[0029] In some embodiments, the droplet quality control device based on laser detection may further include a fluorescence detection device. The position of the fluorescence detection device corresponds to the detection area in the main microchannel and is used to detect whether a target substance exists in the fused droplets in the main microchannel. The target substance is a substance pre-fluorescently labeled, such as nucleic acid, etc.

[0030] Figure 2 is a schematic diagram of the microchannel for generating droplets in a droplet quality control device based on laser detection in an embodiment, and Figure 3 is a schematic diagram of droplet fusion in a droplet quality control device based on laser detection in an embodiment. The schematic diagram shown in this embodiment includes two individual microchannels, the first individual microchannel and the second individual microchannel. The air pressure output by the pressure output device 16 maintains the pressure of the two individual microchannels. Under the action of the pressure, different solutions in each individual microchannel flow in their respective channels and form droplets in their respective individual microchannels. The droplets in each individual microchannel flow to the main microchannel and fuse with each other in the main microchannel to form fused droplets. The fused droplets are as Figure 3 shown.

[0031] Please refer toFigure 4 is a flowchart of a droplet quality control method based on laser detection provided by an embodiment of the present application. The droplet quality control method based on laser detection is applied to a gene sequencer. The droplet quality control method based on laser detection includes the following steps:

[0032] S11. Obtain droplet microchannel signal data collected by a laser device.

[0033] In this embodiment, the laser device can be used to detect a single microchannel or a main microchannel. Therefore, the droplet microchannel includes each single microchannel and / or the main microchannel. The laser device includes a transmitting device and a receiving device. The transmitting device is used to send a laser signal to the laser detection area, and the receiving device receives the signal detected in the laser detection area. Since there are droplets flowing in the droplet microchannel, data such as the size, interval, and flow rate of the droplets will affect the detected signal received. For the laser device used for the single microchannel, the collected signal is the single microchannel signal data; for the laser device used for the main microchannel, the collected signal is the main microchannel signal data. The droplet microchannel signal data includes the single microchannel signal data and / or the main microchannel signal data. If the droplet microchannel includes a single microchannel, the droplets in the droplet microchannel include the droplets in the single microchannel; if the droplet microchannel includes the main microchannel, the droplets in the droplet microchannel include the fused droplets in the single microchannel. Among them, the droplet microchannel signal data can be the signal data collected within a period of time or the signal data collected once.

[0034] S12. Monitor the state of the droplets in the droplet microchannel based on the droplet microchannel signal data.

[0035] In this embodiment, the laser device 13 can collect real-time droplet microchannel signal data during the droplet monitoring process, and can analyze the collected droplet microchannel signal data to monitor the droplet state. The droplet state includes at least one of the following: the droplet conditions in each single microchannel, the fused droplet conditions in the main microchannel, and the generation frequency ratio of each single microchannel. The droplet conditions include but are not limited to droplet size data, droplet interval, and droplet flow rate. The fused droplet conditions include but are not limited to any one of the following: the fusion rate of the droplets, the size data of the fused droplets, the interval of the fused droplets, the flow rate of the fused droplets, the fusion rate of the fused droplets, the encapsulation rate of the fused droplets, whether there is a target substance in the fused droplets, the successful fusion rate of the fused droplets, etc. By determining whether the droplet conditions in each single microchannel and / or the fused droplet conditions generated in the main microchannel meet the preset conditions, it is judged whether there are abnormal problems during the droplet monitoring process.

[0036] In some embodiments, when the droplet conditions in each individual microchannel and / or the conditions of the fused droplets generated in the main microchannel do not meet the preset conditions, it is determined that the state of the droplets in the droplet microchannel is abnormal, and a warning prompt is executed. In an alternative implementation, the ways to execute the warning prompt include, but are not limited to, one or a combination of the following: performing an abnormality prompt on the user interface, sending an abnormality prompt to the terminal devices of relevant personnel. When performing an abnormality prompt on the user interface, images, pressure data, etc. corresponding to the abnormal situation can be displayed.

[0037] In the above embodiments, during the droplet monitoring process, the signal data of the droplet microchannel is collected in real time by a laser device. The signal data of the droplet microchannel includes at least one of the following: the signal data of the individual microchannel, the signal data of the main microchannel. By analyzing the signal data of the individual microchannel and / or the signal data of the main microchannel, the droplet conditions in each individual microchannel and / or the conditions of the fused droplets generated in the main microchannel can be monitored, so that the abnormal states during the entire process of droplet generation, pairing, and fusion can be monitored in real time, facilitating maintenance personnel to promptly discover and eliminate abnormalities.

[0038] In some embodiments, the state of the droplets in the droplet microchannel includes at least one or a combination of the following: the generation frequency of the droplets in each individual microchannel, the current droplet generation frequency relationship corresponding to all individual microchannels, the size data of the droplets in the droplet microchannel, the droplet interval in the droplet microchannel, the flow rate of the droplets in the droplet microchannel, the fusion rate of the droplets in the main microchannel, whether there is a target substance in the fused droplets in the main microchannel, the encapsulation rate of the fused droplets in the main microchannel, whether the fused droplets in the main microchannel are successfully fused droplets, the successful fusion rate of the fused droplets in the main microchannel, the content data of the target substance in the successfully fused droplets.

[0039] In this embodiment, the current droplet generation frequency relationship can be represented by the ratio of the generation frequencies of all individual microchannels. The generation frequency represents the generation speed of the droplets; the fusion rate represents the proportion of the droplets in each individual microchannel that are successfully fused together to form fused droplets; the encapsulation rate indicates the proportion of the fused droplets containing the target substance in the fused droplets; the successfully fused droplets refer to the fused droplets containing the fluorescently labeled target substance; the successful fusion rate represents the proportion of the successfully fused droplets in the fused droplets. The target substance can be a specified substance or a carrier carrying the specified substance. The specified substance includes, but is not limited to, one or a combination of the following: nucleic acids, proteins, carbohydrates, lipids, or other organic or inorganic solvents. For example, the target substance is a gel bead with a specified DNA label.

[0040] In the above embodiments, the process of droplet generation, pairing, and fusion is monitored from multiple angles to promptly discover abnormalities during the monitoring process and ensure the success rate of droplet fusion.

[0041] In some embodiments, monitoring the state of droplets in the droplet microchannel based on the droplet microchannel signal data includes:

[0042] Based on the signal data of each single microchannel, obtaining the pulse signal interval corresponding to each single microchannel, and calculating the generation frequency of droplets in each single microchannel based on the pulse signal intervals corresponding to each single microchannel;

[0043] Calculating the current droplet generation frequency relationship corresponding to all single microchannels according to the generation frequencies of droplets in each single microchannel;

[0044] Controlling the pressure data of each single microchannel based on the current droplet generation frequency relationship.

[0045] In this embodiment, for a single microchannel, in the laser detection area of the laser device, when a droplet passes through the single microchannel, there will be pulse signals in the collected single microchannel signal data. One droplet corresponds to one pulse signal. As Figure 5 shown, Figure 5 is a schematic diagram of the single microchannel signal data in an embodiment. In a single microchannel, when two adjacent droplets pass through the laser detection area, the formed single microchannel signal data, each droplet corresponds to one pulse signal. As Figure 5 shown, the T is a pulse signal interval, and the pulse signal interval indicates the time interval between two adjacent pulse signals. In an alternative implementation, for a single microchannel, obtaining multiple pulse signal intervals, calculating the average value of the pulse signal intervals based on the multiple pulse signal intervals, and taking the average value of the pulse signal intervals as the pulse signal interval corresponding to the single microchannel. The generation frequency of droplets in the single microchannel = 1 / the pulse signal interval corresponding to the single microchannel. The same method can be used to calculate for other single microchannels, so as to obtain the generation frequency of droplets in each single microchannel. The generation frequency represents the generation speed of droplets.

[0046] The current droplet generation frequency relationship represents the ratio of the generation frequencies of all single microchannels. For example, in the case of two single microchannels, the current droplet generation frequency relationship is the ratio of the generation frequency of the first single microchannel to the generation frequency of the second single microchannel. The current droplet generation frequency relationship is related to the fusion rate of droplets. If the droplet generation frequency of a certain single microchannel is too fast or too slow, it will cause waste of reagents and reduce the fusion rate of droplets. Therefore, comparing the current droplet generation frequency relationship with the preset generation frequency ratio to determine whether the current droplet generation frequency relationship meets the preset generation frequency condition, so as to control the pressure data of each single microchannel.

[0047] In an alternative implementation, controlling the pressure data of each single microchannel based on the current droplet generation frequency relationship includes at least one of the following:

[0048] Compare the current droplet generation frequency relationship with a preset generation frequency ratio to obtain single microfluidic channels with a generation frequency lower than a first generation frequency value, and increase the pressure data of the single microfluidic channels with a generation frequency lower than the first generation frequency value;

[0049] Compare the current droplet generation frequency relationship with a preset generation frequency ratio to obtain single microfluidic channels with a generation frequency higher than a second generation frequency value, and reduce the pressure data of the single microfluidic channels with a generation frequency higher than the second generation frequency value.

[0050] In this embodiment, the first generation frequency value and the second generation frequency value are calculated based on a preset generation frequency ratio. By obtaining single microfluidic channels with a generation frequency lower than the first generation frequency value or single microfluidic channels with a generation frequency higher than the second generation frequency value and adjusting the pressure data of these single microfluidic channels, the generation frequencies of these single microfluidic channels can be adjusted, so that the current droplet generation frequency relationship is within the error range of the preset generation frequency ratio, thus ensuring the efficiency of droplet fusion.

[0051] In the above embodiment, the pulse signal intervals corresponding to each single microfluidic channel are obtained. Based on the pulse signal intervals corresponding to each single microfluidic channel, the generation frequencies of the droplets in each single microfluidic channel are calculated, and the current droplet generation frequency relationship corresponding to all single microfluidic channels is calculated. The current droplet generation frequency relationship is compared with the preset generation frequency ratio. When the current droplet generation frequency relationship does not meet the preset generation frequency condition, single microfluidic channels with a generation frequency lower than the first generation frequency value or single microfluidic channels with a generation frequency higher than the second generation frequency value are obtained, and the pressure data of these single microfluidic channels are adjusted, so that the generation frequencies of these single microfluidic channels can be adjusted, and the current droplet generation frequency relationship is within the error range of the preset generation frequency ratio, thus ensuring the success rate of droplet fusion and reducing the waste of reagents.

[0052] In some embodiments, before monitoring the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data, the method further includes:

[0053] Obtain a standard pulse width, which is obtained by placing a standard object with a preset diameter in the droplet microfluidic channel to flow and according to the received signal of the laser device.

[0054] In this embodiment, the standard object can be a standard rubber ball. Placing the standard object in the droplet microfluidic channel to flow, so that when the standard object passes through the laser detection area of the laser device, the corresponding microfluidic channel signal data can be obtained. As Figure 6 shown, Figure 6Schematic diagram of the microchannel signal data corresponding to the standard object in an embodiment. The pulse signals corresponding to the standard rubber balls are collected twice continuously, and one pulse width is represented by T4. In an alternative implementation, multiple pulse signals can be extracted from the microchannel signal data, and the pulse width of each pulse signal can be calculated. The average value after accumulating the pulse widths of each pulse signal is used as the standard pulse width. Since the standard object has a preset diameter, the interval between droplets, size data, etc. can be estimated based on the standard pulse width. The standard pulse width indicates the average width of the pulse signal corresponding to the standard object.

[0055] In the above embodiment, by obtaining the standard pulse width corresponding to the standard object with a preset diameter, the relevant data of the droplets can be accurately estimated subsequently, so as to accurately monitor the state during the droplet monitoring process and detect abnormalities in a timely manner.

[0056] In some embodiments, the monitoring of the droplets in the droplet microchannel based on the droplet microchannel signal data includes:

[0057] Based on the droplet microchannel signal data, obtain at least one pulse width corresponding to the droplet microchannel;

[0058] Based on the multiple pulse widths, the preset diameter, and the standard pulse width, calculate the size data of the droplets in the droplet microchannel;

[0059] Based on the size data of the droplets, perform an abnormal warning operation.

[0060] In this embodiment, this step can be used to calculate the size data of the droplets in a single microchannel or the size data of the fused droplets in the main microchannel. For a single microchannel or the main microchannel, at least one pulse width can be obtained from the single microchannel signal data or the main microchannel signal data. For each pulse width, based on the preset diameter and the standard pulse width, a droplet size can be obtained, that is, the size data of the droplet. If one pulse width is T1, the preset diameter is D, and the standard pulse width is T5, then a droplet size d = D * T1 / T5. Therefore, each pulse width corresponds to a droplet size. Based on the size data of the droplets, determine whether the droplets in the single microchannel and / or the main microchannel meet the preset conditions. When it is determined based on the size data of the droplets that the droplets in the single microchannel and / or the main microchannel do not meet the preset conditions, perform an abnormal warning operation.

[0061] In an alternative implementation, performing an abnormal warning operation based on the size data of the droplets includes:

[0062] For a single microchannel, obtain size data of multiple droplets; calculate the average droplet size based on the size data of the multiple droplets, and when the average droplet size does not meet the preset average droplet size, perform an abnormal warning operation; or

[0063] For the main microchannel, screen out unmerged target droplets according to the size data of the droplets, calculate the number of target droplets, and perform an abnormal warning operation based on the number of target droplets.

[0064] In the above embodiments, during the monitoring of droplets, the size data of droplets in the single microchannel and / or the main microchannel can be monitored. Based on the size data of the droplets, when it is determined that the droplets in the single microchannel and / or the main microchannel do not meet the preset conditions, an abnormal warning operation is performed, so as to accurately monitor the abnormalities during the droplet monitoring process, improve the success rate of droplet fusion, and reduce the waste of reagent resources.

[0065] In some embodiments, the monitoring of the state of droplets in the droplet microchannel based on the droplet microchannel signal data includes:

[0066] Based on the droplet microchannel signal data, obtain the pulse signal interval corresponding to the droplet microchannel;

[0067] Based on the pulse signal interval corresponding to the droplet microchannel, the preset diameter, and the standard pulse width, calculate the droplet interval in the droplet microchannel;

[0068] Based on the droplet interval in the droplet microchannel, perform an abnormal warning operation.

[0069] In this embodiment, this step can be used to calculate the size data of droplets in a single microchannel or the size data of the fused droplets in the main microchannel. For a single microchannel or the main microchannel, calculating the pulse signal interval corresponding to the single microchannel or the main microchannel is similar to the method of obtaining the pulse signal interval corresponding to the single microchannel described in the above embodiment, and will not be elaborated here. If the pulse signal interval corresponding to the single microchannel or the main microchannel is T2, the preset diameter is D, and the standard pulse width is T5, then the droplet interval d3 in the single microchannel or the main microchannel = D * T2 / T5. In an alternative implementation, the pulse signal interval corresponding to the droplet microchannel may not be the average pulse signal interval, but represent multiple pulse signal intervals corresponding to the droplet microchannel. For one pulse signal interval, using the above formula, the droplet interval corresponding to this pulse signal interval can be calculated, so as to obtain the droplet intervals corresponding to multiple pulse signal intervals. Based on the droplet intervals corresponding to multiple pulse signal intervals, the average droplet interval can be obtained. In an alternative implementation, based on the droplet interval in the droplet microchannel, performing the abnormal warning operation includes at least one of the following: when the average droplet interval in the single microchannel or the main microchannel is not within the error range of the preset droplet interval, performing the abnormal warning operation; or when the number of droplet intervals in the single microchannel or the main microchannel that are not within the error range of the preset droplet interval exceeds the preset number, performing the abnormal warning operation. When the droplet intervals in the single microchannel or the main microchannel are uneven, it indicates that the pressure in the single microchannel or the main microchannel is unstable, and the cause needs to be checked in time.

[0070] In the above embodiment, during the monitoring of droplets, the droplet intervals in the single microchannel and / or the main microchannel can be monitored. Based on the droplet intervals, when it is determined that the droplet intervals in the single microchannel and / or the main microchannel do not meet the preset droplet interval conditions, an abnormal warning operation is performed, so as to accurately monitor the abnormalities during the droplet monitoring process, improve the success rate of droplet fusion, and reduce the waste of reagent resources.

[0071] In some embodiments, the monitoring of the state of droplets in the droplet microchannel based on the droplet microchannel signal data includes:

[0072] Calculating the flow rate of droplets in the droplet microchannel according to the preset diameter and the standard pulse width;

[0073] When the flow rate of droplets in the droplet microchannel does not meet the preset flow rate conditions, an abnormal warning operation is performed.

[0074] In this embodiment, since the various parameter conditions of the standard object are the same during the droplet microchannel and droplet monitoring processes, the flow rate of the droplet can be represented by the flow rate of the standard object. The droplet flow rate S = preset diameter / standard pulse width. The droplet flow rate cannot be too fast or too slow, as being too fast or too slow will affect the droplet fusion rate in the main microchannel. In an alternative implementation, when the flow rate of the droplet in the droplet microchannel does not meet the preset flow rate condition, the abnormal warning operation includes one of the following: when the flow rate of the droplet is lower than the first preset flow rate value, an abnormal warning operation is performed to indicate that the flow rate is too slow; when the flow rate of the droplet is higher than the second preset flow rate value, an abnormal warning operation is performed to indicate that the flow rate is too fast; where the second preset flow rate value is greater than the first preset flow rate value.

[0075] In the above embodiment, during the monitoring of the droplet, the flow rate of the droplet in the droplet microchannel can be monitored. Based on the droplet flow rate, when it is determined that the flow rate of the droplet in the droplet microchannel does not meet the preset flow rate condition, an abnormal warning operation is performed, thereby accurately monitoring the abnormality during the droplet monitoring process, improving the success rate of droplet fusion, and reducing the waste of reagent resources.

[0076] In some embodiments, the monitoring of the state of the droplet in the droplet microchannel based on the droplet microchannel signal data includes:

[0077] Obtaining the total number of droplets flowing in the target single microchannel within the target time period, and obtaining the target droplets in the main microchannel whose droplet sizes are within a preset range of the droplet sizes in the target single microchannel during the monitoring time period corresponding to the target time period, and calculating the number of target droplets;

[0078] Calculating the fusion rate based on the number of target droplets and the total number of droplets;

[0079] When the fusion rate is lower than the preset fusion rate, an abnormal warning operation is performed.

[0080] In this embodiment, the target single microchannel can be any single microchannel. The monitoring time period is used for the time period in the main microchannel. Since the droplets in the single microchannel need to flow into the main microchannel first and then the droplets are fused, the monitoring time period will be later than the target time period. The monitoring time period is used to monitor the time period of the main microchannel, and the target time period is used to monitor the time period of the target single microchannel. In an alternative implementation, the total number of droplets in the target time period can be determined from the number of pulse signals in the signal data of the target single microchannel in the target time period. Generally, the size data of the fused droplets is larger than the size data of the droplets in the single microchannel. Therefore, target droplets with droplet sizes within a preset range between the fused channel in the main microchannel and the droplets in the target single microchannel can be obtained, and the target droplets are the unfused droplets. For example, if the total number of droplets in the target time period is 100 and the number of target droplets in the monitoring time period is 20, then the number of droplets participating in fusion is 80, and the fusion rate is 80 / 100. That is, the fusion rate is (total number of droplets - number of target droplets) / total number of droplets. The fusion rate represents the proportion of droplets in each single microchannel that are successfully fused together to form fused droplets.

[0081] In the above embodiment, during the monitoring of droplets, the fusion rate of the droplets in the main microchannel can be monitored. When the fusion rate is lower than the preset fusion rate, an abnormal warning operation is performed, so as to accurately monitor the abnormalities during the droplet monitoring process, improve the success rate of droplet fusion, and reduce the waste of reagent resources.

[0082] In some embodiments, the monitoring of the state of droplets in the droplet microchannel based on the droplet microchannel signal data includes:

[0083] Obtain the main microchannel signal data, and extract the pulse signals corresponding to each droplet in the main microchannel from the main microchannel signal data;

[0084] Determine the fused droplets with target substances as the droplets whose signal attributes of the corresponding pulse signals in the main microchannel meet the preset signal attribute conditions; where the signal attributes include at least one of the following: there is a reverse trough between the positions of two wave peaks in the pulse signal, and the pulse width is greater than the pulse width of the pulse signal corresponding to the droplets in the single microchannel.

[0085] In this embodiment, the signal data corresponding to the fused droplets can be collected through the laser detection area corresponding to the main microchannel. Since it is possible that after droplet fusion, there is no DNA tag microsphere encapsulated in the droplet. If there is no DNA tag microsphere encapsulated in the droplet, accurate gene data cannot be detected subsequently. Therefore, it is necessary to monitor whether the fused droplets in the main microchannel are droplets encapsulated with tags. As Figure 7 shown, Figure 7 is a schematic diagram of the pulse signal of the fused droplets with target substances in an embodiment, and the reverse trough is asFigure 7 as shown; relative to Figure 5 as shown, an inverted peak will appear in the middle of the pulse waveform of the label-wrapped fusion droplet, and the waveform becomes wider, and the peak of the label-wrapped fusion droplet is higher than the peak of the pulse signal corresponding to the droplet in the single microchannel.

[0086] In an alternative embodiment, the monitoring of the state of the droplets in the droplet microchannel based on the droplet microchannel signal data further includes:

[0087] Obtaining the total number of fusion droplets in the main microchannel and obtaining the number of fusion droplets with the target;

[0088] Calculating the encapsulation rate according to the total number of fusion droplets in the main microchannel and the number of fusion droplets with the target;

[0089] When the encapsulation rate is lower than the preset encapsulation value, an abnormal warning operation is performed.

[0090] In this embodiment, the total number of fusion droplets in the main microchannel within a preset time period can be monitored. The signal data of the main microchannel within the preset time period can be preset. According to the signal data of the main microchannel, the size data of each droplet in the main microchannel is obtained. According to the size data of each droplet in the main microchannel, the fusion droplets are determined. For example, the droplets with size data higher than the preset droplet size data are determined as fusion droplets, and then the total number of fusion droplets is calculated. The encapsulation rate is the number of fusion droplets with the target / the total number of fusion droplets. The encapsulation rate indicates the proportion of fusion droplets with the target in the fusion droplets.

[0091] In the above embodiment, during the monitoring of the droplets, the encapsulation rate of the droplets in the main microchannel can be monitored. When the fusion rate is lower than the preset encapsulation value, an abnormal warning operation is performed, so as to accurately monitor the abnormality during the droplet monitoring process, improve the success rate of droplet fusion, and reduce the waste of reagent resources.

[0092] In some embodiments, the monitoring of the state of the droplets in the droplet microchannel based on the droplet microchannel signal data includes at least one of:

[0093] Obtaining the signals corresponding to each fusion droplet in the main microchannel through a fluorescence detection device, determining the fusion droplets corresponding to the signals with fluorescence signals as successfully fused droplets, obtaining the total number of the successfully fused droplets and obtaining the number of fusion droplets in the main microchannel, calculating the successful fusion rate according to the total number of the successfully fused droplets and the number of fusion droplets in the main microchannel, and performing an abnormal warning operation when the successful fusion rate is lower than the preset successful fusion rate; or

[0094] Determine the content data of the target substance labeled with a fluorescent dye in the successfully fused droplet according to the fluorescence signal intensity of the successfully fused droplet.

[0095] In this embodiment, after the droplets in each single microchannel in the main microchannel are fused, fused droplets are obtained. However, there may be some droplets that are fused but not successfully fused. Certain substances can be pre-labeled with fluorescence in advance, and whether there is a fluorescence signal in the signal of the fused droplet is detected by a fluorescence detection device. If there is a fluorescence signal, it indicates a successfully fused droplet. Therefore, fused droplets can be screened out first through the size data of the droplets, and then whether the corresponding signal of the fused droplet has a fluorescence signal is detected. Specifically, the fluorescence detection device emits light of a certain wavelength to the fluorescence detection area of the main microchannel. The fused droplet with a fluorescence label is excited to emit a fluorescence signal of a specific wavelength. If a fluorescence signal of a specific wavelength is detected, it indicates that there is a fluorescence signal in the signal of the fused droplet, that is, the droplet is successfully fused. The successful fusion rate represents the proportion of successfully fused droplets in the fused droplets. In some embodiments, the content data of the target substance in the successfully fused droplet can also be determined according to the intensity value of the fluorescence signal, where the content data includes but is not limited to concentration, quantity, proportion, etc.

[0096] In the above embodiment, during the monitoring process of the droplets, the successful fusion rate of the droplets in the main microchannel can be monitored. When the fusion rate is lower than the preset successful fusion rate, an abnormal warning operation is performed. The content of the target substance in the successfully fused droplet can also be monitored to ensure that the droplets entering the subsequent process are reliable droplets.

[0097] In some embodiments, the method further includes:

[0098] Obtain droplet microchannel image information, where the droplet microchannel image information includes single microchannel image information and / or main microchannel image information;

[0099] Based on the droplet microchannel image information, monitor the state of the droplets in the droplet microchannel.

[0100] In this embodiment, the single microchannel image information and / or the main microchannel image information can be analyzed to monitor the size data, interval, morphology, etc. of the droplets in the single microchannel. In combination with one or more of the above embodiments, the state of the droplets in the droplet microchannel is monitored. When the state of the droplets monitored based on the droplet microchannel signal data is inconsistent with the state of the droplets monitored based on the droplet microchannel image information, an abnormal warning operation can be performed, and the obtained droplet microchannel image is sent to the terminal device for the user to make a final confirmation.

[0101] In the above embodiments, by combining real-time image analysis and laser detection, the processes of droplet generation, pairing, fusion, etc. are monitored in real time from different angles, thereby improving the accuracy and success rate of the experiment.

[0102] On the other hand, the present application provides a computer program product, including a computer program, which when executed by a processor implements the droplet quality control method based on laser detection according to any embodiment of the present application.

[0103] Among them, in the computer program product, an optional implementation form of the program module architecture of the computer program for implementing each step of the method can be a droplet quality control device based on laser detection. Please refer to Figure 8 , an embodiment of the present application provides a droplet quality control device based on laser detection, including: an acquisition module 81 for acquiring droplet microchannel signal data collected by a laser device, and a monitoring module 82 for monitoring the state of droplets in the droplet microchannel based on the droplet microchannel signal data; wherein the droplet microchannel includes each single microchannel and / or the main microchannel, the droplets in the droplet microchannel include the droplets in each single microchannel and / or the fused droplets in the main microchannel, the droplet microchannel signal data includes single microchannel signal data and / or main microchannel signal data, after the solutions corresponding to each single microchannel form the droplets in each single microchannel in their respective corresponding single microchannels, the droplets in each single microchannel flow to the main microchannel and fuse in the main microchannel to form fused droplets.

[0104] Optionally, the state of the droplets in the droplet microchannel includes at least one or a combination of the following: the generation frequency of droplets in each single microchannel, the current droplet generation frequency relationship corresponding to all single microchannels, the size data of the droplets in the droplet microchannel, the droplet interval in the droplet microchannel, the flow rate of the droplets in the droplet microchannel, the fusion rate of droplets in the main microchannel, whether there is a target substance in the fused droplets in the main microchannel, the encapsulation rate of the fused droplets in the main microchannel, whether the fused droplets in the main microchannel are successfully fused droplets, the successful fusion rate of the fused droplets in the main microchannel, and the content data of the target substance in the successfully fused droplets.

[0105] Optionally, the monitoring module 82 is further configured to:

[0106] Based on the single microchannel signal data of each single microchannel, obtain the pulse signal interval corresponding to each single microchannel, and calculate the generation frequency of the droplets in each single microchannel based on the pulse signal interval corresponding to each single microchannel;

[0107] Calculate the current droplet generation frequency relationship corresponding to all single microchannels according to the generation frequency of the droplets in each single microchannel;

[0108] Based on the current droplet generation frequency relationship, control the pressure data of each single microchannel.

[0109] Optionally, the monitoring module 82 is further configured to:

[0110] Compare the current droplet generation frequency relationship with a preset generation frequency ratio, obtain the single microchannels with a generation frequency lower than the first generation frequency value, and increase the pressure data of the single microchannels with a generation frequency lower than the first generation frequency value;

[0111] Compare the current droplet generation frequency relationship with a preset generation frequency ratio, obtain the single microchannels with a generation frequency higher than the second generation frequency value, and reduce the pressure data of the single microchannels with a generation frequency higher than the second generation frequency value.

[0112] Optionally, the monitoring module 82 is further configured to:

[0113] Obtain a standard pulse width, where the standard pulse width is obtained by placing a standard object with a preset diameter in the droplet microchannel to flow and according to the received signal of the obtained laser device.

[0114] Optionally, the monitoring module 82 is further configured to:

[0115] Based on the droplet microchannel signal data, obtain at least one pulse width corresponding to the droplet microchannel;

[0116] Based on the multiple pulse widths, the preset diameter, and the standard pulse width, calculate the size data of the droplets in the droplet microchannel;

[0117] Based on the size data of the droplets, perform an abnormal warning operation.

[0118] Optionally, the monitoring module 82 is further configured to:

[0119] Based on the droplet microchannel signal data, obtain the pulse signal interval corresponding to the droplet microchannel;

[0120] Based on the pulse signal interval corresponding to the droplet microchannel, the preset diameter, and the standard pulse width, calculate the droplet interval in the droplet microchannel;

[0121] Based on the droplet interval in the droplet microchannel, perform an abnormal warning operation.

[0122] Optionally, the monitoring module 82 is further configured to:

[0123] According to the preset diameter and the standard pulse width, calculate the flow rate of the droplets in the droplet microchannel;

[0124] When the flow rate of the droplets in the droplet microchannel does not meet the preset flow rate condition, perform an abnormal warning operation.

[0125] Optionally, the monitoring module 82 is further configured to:

[0126] Obtain the total number of droplets flowing in the target single microchannel within the target time period, and obtain target droplets in the main microchannel whose droplet sizes are within a preset range from the droplet sizes in the target single microchannel during the monitoring time period corresponding to the target time period, and calculate the number of target droplets;

[0127] Calculate a fusion rate based on the number of target droplets and the total number of droplets;

[0128] When the fusion rate is lower than a preset fusion rate, perform an abnormal warning operation.

[0129] Optionally, the monitoring module 82 is further configured to:

[0130] Obtain main microchannel signal data, and extract pulse signals corresponding to each droplet in the main microchannel from the main microchannel signal data;

[0131] Determine droplets whose signal attributes of the corresponding pulse signals in the main microchannel meet preset signal attribute conditions as fusion droplets with a target substance; where the signal attributes include at least one of the following: there is a reverse trough between the positions of two wave peaks in the pulse signal, and the pulse width is greater than the pulse width of the pulse signal corresponding to the droplet in the single microchannel.

[0132] Optionally, the monitoring module 82 is further configured to:

[0133] Obtain the total number of fusion droplets in the main microchannel, and obtain the number of fusion droplets with a target substance;

[0134] Calculate a wrapping rate based on the total number of fusion droplets in the main microchannel and the number of fusion droplets with a target substance;

[0135] When the wrapping rate is lower than a preset wrapping value, perform an abnormal warning operation.

[0136] Optionally, the monitoring module 82 is further configured to:

[0137] Obtain signals corresponding to each fusion droplet in the main microchannel through a fluorescence detection device, determine the fusion droplets corresponding to the signals with fluorescence signals as successfully fused droplets, obtain the total number of the successfully fused droplets and obtain the number of fusion droplets in the main microchannel, calculate a successful fusion rate based on the total number of the successfully fused droplets and the number of fusion droplets in the main microchannel, and when the successful fusion rate is lower than a preset successful fusion rate, perform an abnormal warning operation; or

[0138] Based on the fluorescence signal intensity of the successfully fused droplets, determine the content data of the target substance labeled with the fluorescent dye in the successfully fused droplets.

[0139] Optionally, the monitoring module 82 is further configured to:

[0140] Obtain droplet microchannel image information, where the droplet microchannel image information includes single microchannel image information and / or main microchannel image information;

[0141] Based on the droplet microchannel signal data and the droplet microchannel image information, monitor the state of the droplets in the droplet microchannel.

[0142] Those skilled in the art can understand that Figure 8 the structure of the droplet quality control device based on laser detection does not constitute a limitation to the droplet quality control device based on laser detection, and the various modules can be implemented in whole or in part by software, hardware, and their combinations. The above-mentioned modules can be embedded in the controller of the computer device in hardware form or be independent of it, or can be stored in the memory of the computer device in software form, so that the controller can call and execute the operations corresponding to the above-mentioned modules. In other embodiments, the droplet quality control device based on laser detection may include more or fewer modules than shown in the figure.

[0143] Please refer to Figure 9 , on the other hand, an embodiment of the present application further provides a droplet quality control device 200 based on laser detection, including a controller 10 and a memory 18. The memory 18 stores a computer program, and when the computer program is executed by the controller, the controller 10 executes the steps of the droplet quality control method provided in any one of the above embodiments of the present application.

[0144] Among them, the controller 10 is the control center, which connects various parts of the entire computer device through various interfaces and lines. By running or executing the software programs and / or modules stored in the memory 18, and by calling the data stored in the memory 18, it executes various functions of the computer device and processes data. Optionally, the controller 10 may include one or more processing cores; preferably, the controller 10 may integrate an application controller and a modulation and demodulation controller. Among them, the application controller mainly processes the operating system, user interfaces, and application programs, etc., and the modulation and demodulation controller mainly processes wireless communications. It can be understood that the above-mentioned modulation and demodulation controller may not be integrated into the controller 10.

[0145] The memory 18 can be used to store software programs and modules. By running the software programs and modules stored in the memory 18, the controller 10 can execute various functional applications and data processing. The memory 18 may mainly include a program storage area and a data storage area. Among them, the program storage area can store an operating system, application programs required for at least one function (such as a sound playback function, an image playback function, etc.); the data storage area can store data created according to the use of the computer device. In addition, the memory 18 can include a high-speed random access memory, and can also include a non-volatile memory, such as at least one magnetic disk storage device, a flash memory device, or other non-volatile solid-state storage devices. Accordingly, the memory 18 can also include a memory controller to provide the controller 10 with access to the memory 18.

[0146] In another aspect of the embodiments of the present application, a storage medium is further provided, storing a computer program, which, when executed by the controller, causes the controller to execute the steps of the droplet quality control method based on laser detection provided in any one of the above embodiments of the present application.

[0147] In another aspect of the embodiments of the present application, a single-cell sequencing device is further provided, including the droplet quality control device based on laser detection provided in any one of the above embodiments of the present application.

[0148] In another aspect of the embodiments of the present application, a single-cell sequencing library construction device is further provided, including the droplet quality control device based on laser detection provided in any one of the above embodiments of the present application.

[0149] Those of ordinary skill in the art can understand that all or part of the processes in the methods provided in the above embodiments can be completed by instructing relevant hardware through a computer program. The program can be stored in a non-volatile computer-readable storage medium. When the program is executed, it can include the processes of the embodiments of the above methods. Among them, any reference to a memory, storage, database, or other medium used in the embodiments provided in the present application can include non-volatile and / or volatile memories. Non-volatile memories can include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or flash memory. Volatile memories can include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in various forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDR SDRAM), enhanced SDRAM (ESDRAM), synchronous link DRAM (SLDRAM), Rambus direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and Rambus dynamic RAM (RDRAM), etc.

[0150] As described above, the above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention can easily think of changes or substitutions, which should all be covered by the protection scope of the present invention. The protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. A droplet quality control method based on laser detection, characterized in that: include: Acquire the droplet microfluidic signal data collected by the laser device, Based on the droplet microfluidic channel signal data, the state of droplets in the droplet microfluidic channel is monitored; wherein the droplet microfluidic channel includes each single microfluidic channel and a main microfluidic channel, the droplets in the droplet microfluidic channel include droplets in each single microfluidic channel and fused droplets in the main microfluidic channel, the droplet microfluidic channel signal data includes each single microfluidic channel signal data and the main microfluidic channel signal data, after the solutions corresponding to each single microfluidic channel form droplets in each single microfluidic channel in their respective corresponding single microfluidic channels, the droplets in each single microfluidic channel flow to the main microfluidic channel and merge in the main microfluidic channel to form fused droplets, wherein the state of droplets in the droplet microfluidic channel includes whether there is a target object in the fused droplets in the main microfluidic channel and the encapsulation rate of the fused droplets in the main microfluidic channel; wherein based on the droplet microfluidic channel signal data, the droplets in the droplet microfluidic channel are monitored. The state of droplets in the droplet microchannel includes: obtaining main microchannel signal data, and extracting pulse signals corresponding to each droplet in the main microchannel from the main microchannel signal data; determining droplets whose signal attributes of the corresponding pulse signal in the main microchannel meet preset signal attribute conditions as fused droplets with target objects; wherein the signal attributes include at least one of the following: there is a reverse trough between two peak positions in the pulse signal, and the pulse width is greater than the pulse width of the pulse signal corresponding to the droplets in a single microchannel; obtaining the total number of fused droplets in the main microchannel, and obtaining the number of fused droplets with target objects; calculating the encapsulation rate according to the total number of fused droplets in the main microchannel and the number of fused droplets with target objects; when the encapsulation rate is lower than the preset encapsulation value, performing an abnormal warning operation.

2. The droplet quality control method based on laser detection according to claim 1, characterized in that: The state of droplets in the droplet microfluidic channel includes at least one of the following: the generation frequency of droplets in each single microfluidic channel, the relationship between the current droplet generation frequencies corresponding to all single microfluidic channels, the size data of droplets in the droplet microfluidic channel, the droplet spacing in the droplet microfluidic channel, the flow rate of droplets in the droplet microfluidic channel, the fusion rate of droplets in the main microfluidic channel, whether the fused droplets in the main microfluidic channel are successfully fused droplets, the successful fusion rate of fused droplets in the main microfluidic channel, and the content data of the target substance in the successfully fused droplets.

3. The droplet quality control method based on laser detection according to claim 2, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data comprises: Based on the signal data of each single microchannel, the pulse signal interval corresponding to each single microchannel is obtained, and based on the pulse signal interval corresponding to each single microchannel, the generation frequency of droplets in each single microchannel is calculated; According to the generation frequency of droplets in each single microfluidic channel, the current droplet generation frequency relationship corresponding to all single microfluidic channels is calculated; Based on the current droplet generation frequency relationship, the pressure data of each single microfluidic channel is controlled.

4. The droplet quality control method based on laser detection according to claim 3, characterized in that: The controlling of the pressure data of each single microchannel based on the current droplet generation frequency relationship includes at least one of the following: Compare the current droplet generation frequency relationship with a preset generation frequency ratio, obtain a single microchannel with a generation frequency lower than a first generation frequency value, and increase the pressure data of the single microchannel with a generation frequency lower than the first generation frequency value; The current droplet generation frequency relationship is compared with a preset generation frequency ratio, a single microchannel having a generation frequency higher than a second generation frequency value is obtained, and pressure data of the single microchannel having a generation frequency higher than the second generation frequency value is reduced.

5. The droplet quality control method based on laser detection according to claim 2, characterized in that: Before monitoring the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data, the method further includes: A standard pulse width is obtained, where the standard pulse width is obtained by placing a standard object with a preset diameter in the droplet microchannel and flowing the standard pulse width according to a received signal of the laser device.

6. The droplet quality control method based on laser detection according to claim 5, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data comprises: Based on the droplet microfluidic channel signal data, acquiring at least one pulse width corresponding to the droplet microfluidic channel; Calculating size data of a droplet in the droplet microfluidic channel based on a plurality of pulse widths, the preset diameter, and the standard pulse width; Based on the droplet size data, abnormal warning operations are performed.

7. The droplet quality control method based on laser detection according to claim 5, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data comprises: Based on the droplet microfluidic channel signal data, obtaining a pulse signal interval corresponding to the droplet microfluidic channel; Calculating the droplet interval in the droplet microchannel based on the pulse signal interval corresponding to the droplet microchannel, the preset diameter and the standard pulse width; Based on the droplet interval in the droplet micro-channel, an abnormality warning operation is performed.

8. The droplet quality control method based on laser detection according to claim 5, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data comprises: Calculating the flow rate of the droplet in the droplet microchannel according to the preset diameter and the standard pulse width; When the flow rate of the droplets in the droplet microchannel does not meet the preset flow rate condition, an abnormal warning operation is performed.

9. The droplet quality control method based on laser detection according to claim 2, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data comprises: Obtaining the total amount of droplets flowing in the target single microfluidic channel within a target time period, and obtaining target droplets whose droplet sizes in the main microfluidic channel and the droplet sizes in the target single microfluidic channel are within a preset range within a monitoring time period corresponding to the target time period, and calculating the number of target droplets; Calculating a fusion rate based on the target number of droplets and the total amount of droplets; When the fusion rate is lower than the preset fusion rate, an abnormal warning operation is performed.

10. The droplet quality control method based on laser detection according to claim 2, characterized in that: The monitoring of the state of the droplets in the droplet microfluidic channel based on the droplet microfluidic channel signal data includes at least one of: Acquire signals corresponding to each fused droplet in the main microfluidic channel by means of a fluorescence detection device, determine the fused droplet corresponding to the signal with the fluorescence signal as a successfully fused droplet, acquire the total number of the successfully fused droplets and the number of fused droplets in the main microfluidic channel, calculate the successful fusion rate according to the total number of the successfully fused droplets and the number of fused droplets in the main microfluidic channel, and perform an abnormal warning operation when the successful fusion rate is lower than a preset successful fusion rate; or According to the fluorescence signal intensity of the successfully fused droplet, the content data of the target substance labeled with the fluorescent dye in the successfully fused droplet is determined.

11. The droplet quality control method based on laser detection according to any one of claims 1 to 10, characterized in that: The method further comprises: Acquiring droplet microfluidic channel image information, wherein the droplet microfluidic channel image information includes single microfluidic channel image information and / or main microfluidic channel image information; Based on the droplet microfluidic channel signal data and the droplet microfluidic channel image information, the state of the droplets in the droplet microfluidic channel is monitored.

12. A droplet quality control device based on laser detection, characterized in that: The method comprises a memory and a controller, wherein the memory stores a computer program, and when the computer program is executed by the controller, the controller executes the steps of the method according to any one of claims 1 to 11.

13. A single-cell sequencing library construction device, characterized in that: Including a droplet quality control device based on laser detection as described in claim 12.

14. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a computer program, and when the computer program is executed by the controller, the droplet quality control method based on laser detection as described in any one of claims 1 to 11 is implemented.

15. A computer program product comprising a computer program, characterized in that When the computer program is executed by the controller, the droplet quality control method based on laser detection as described in any one of claims 1 to 11 is implemented.

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