Signal synchronous acquisition method, microfluidic cell sorter and storage medium

Abstract

The invention relates to the technical field of droplet microfluidics, and discloses a signal synchronous acquisition method, a microfluidic cell sorter and a storage medium wherein the signal synchronous acquisition method comprises the following steps: obtaining a position offset delta X of a fluorescence detection area relative to an image detection area at a preset droplet flow rate, the position offset delta X is related to the image processing time of the image data; adjusting the position of a fluorescence detection area; image data and fluorescence signals of the liquid drops at the preset liquid drop flow speed are synchronously collected. The microfluidic cell sorter comprises a camera, an adjustable excitation light spot module, a PMT and a control unit, wherein the control unit controls starting of the camera and the PMT. A computer program is stored in the storage medium. When a computer program is operated by a processor, the signal synchronous acquisition method is executed. The technical effect of synchronously collecting image data and fluorescence signals is achieved.

Description

Technical Field

[0001] This invention belongs to the field of droplet microfluidics technology, specifically relating to a signal synchronous acquisition method, a microfluidic cell sorter, and a storage medium. Background Technology

[0002] Droplet microfluidics is a technology that uses microreactors formed by encapsulating the sample or reaction system within tiny droplets as independent operating units. It achieves droplet generation, manipulation, detection, and sorting by precisely controlling the behavior of fluids within microchannels. Microfluidic cell sorters are novel analytical instruments based on droplet microfluidics. They generate monodisperse droplets using a microfluidic chip, encapsulate the sample within the droplets, and drive the droplets along microchannels, performing qualitative and quantitative analysis of the sample in the process.

[0003] Currently, microfluidic cell sorters use cameras to acquire images of cells or particles within droplets and collect fluorescence signals via PMTs for multi-parameter phenotypic analysis. To address the issue of asynchronous image data acquisition and fluorescence signal acquisition, microfluidic cell sorters typically employ a single hardware clock to synchronize camera frame rate and fluorescence acquisition.

[0004] However, cameras acquire data frame by frame, while PMTs acquire data in continuous waveform units, leading to a difference in clock resolution. Furthermore, after capturing images, the camera requires image processing time, while the PMT's acquisition and processing time for fluorescence signals is extremely short. This creates a discrepancy between the image data processing time and the fluorescence signal processing time, which is further amplified by changes in droplet flow rate. Therefore, image data and fluorescence data cannot perfectly correspond within the same droplet, easily leading to mismatches and misjudgments, ultimately reducing the accuracy and reliability of the microfluidic cell sorter. Summary of the Invention

[0005] To address the shortcomings of the prior art, this invention provides a signal synchronous acquisition method, a microfluidic cell sorter, and a storage medium. By adjusting the position of the fluorescence detection area, the image acquisition and fluorescence signal acquisition can be aligned in time, ensuring that the image data and fluorescence signal correspond completely within the same droplet, thereby improving the resolution and analysis accuracy of the microfluidic cell sorter.

[0006] The technical effects to be achieved by this invention are realized through the following technical aspects:

[0007] In a first aspect, the present invention provides a signal synchronous acquisition method, comprising the following steps: an image detection area for acquiring image data and a fluorescence detection area for exciting and acquiring fluorescence signals are provided on a microfluidic chip; the positional offset ΔX of the fluorescence detection area relative to the image detection area at a preset droplet flow rate is obtained, wherein the positional offset ΔX is related to the image processing time of the image data; the position of the fluorescence detection area is adjusted such that the distance between the fluorescence detection area and the image detection area is the positional offset ΔX; a droplet flows through the image detection area and the fluorescence detection area, and image data and fluorescence signals of the droplet are synchronously acquired at the preset droplet flow rate.

[0008] In some implementations, the offset ΔX is calculated using the formula: ΔX = Vdroplet Tproc, where Vdroplet is the droplet flow rate and Tproc is the image processing time.

[0009] In some implementations, the droplet velocity Vdroplet is obtained by processing image data of the droplets within the image detection area.

[0010] In some implementations, the formula for calculating the droplet velocity Vdroplet is:

[0011] Where S is the relative displacement of the droplet in the first frame image and the droplet in the second frame image acquired from the image detection area, and Tcam is the sampling period of each frame image.

[0012] In some implementations, the sampling period Tcam of each frame of the camera is calculated using the following formula:

[0013] Where F is the frame rate, and image data of droplets in the image detection area are acquired based on the frame rate F.

[0014] In a second aspect, the present invention provides a microfluidic cell sorting instrument for simultaneously acquiring image data and fluorescence signals of droplets on a microfluidic chip, wherein the microfluidic chip is provided with an image detection area and a fluorescence detection area, and the microfluidic cell sorting instrument includes: A camera is used to acquire images of droplets flowing through the image detection area; An adjustable excitation spot module is used to project an excitation spot within the fluorescence detection area and adjust the projection position of the excitation spot. PMT, used to acquire the fluorescence signal of the droplets within the fluorescence detection area; and The control unit is electrically connected to the camera and the PMT respectively. The control unit executes the signal synchronization acquisition method as described above so that the image data of the droplet and the fluorescence signal are acquired synchronously.

[0015] In some implementations, the adjustable excitation spot module includes a spatial light modulator that adjusts the position of the excitation spot according to an offset ΔX.

[0016] In some implementations, a feedback subsystem is included, which measures the droplet flow velocity Vdroplet in real time after the camera acquires an image of the droplet in order to dynamically adjust the offset ΔX of the excitation spot.

[0017] In some implementations, the microfluidic chip includes a channel for droplet flow, with the image detection area and fluorescence detection area located within the channel. The channel is provided with a droplet generation area and a transport channel, where the droplet is generated in the droplet generation area and flows along the transport channel to the image detection area and the fluorescence detection area.

[0018] Thirdly, the present invention provides a computer-readable storage medium on which a computer program is stored; the computer program is executed by a processor to perform the signal synchronization acquisition method described above.

[0019] In summary, the present invention has at least the following advantages: 1. The signal synchronization acquisition method provided by the present invention adjusts the position of the fluorescence detection area so that the fluorescence detection area is offset by a certain distance relative to the image detection area. The time it takes for the droplet to flow through this distance at a preset flow rate can provide response time for the acquisition and processing of image data. This allows the fluorescence signal in the fluorescence detection area to be processed synchronously when the camera finishes processing the image data. The time window for image data acquisition coincides with the time window for fluorescence signal acquisition, ensuring data synchronization.

[0020] During calibration, the positional offset ΔX of the fluorescence detection area relative to the image detection area is first determined. This offset ΔX is related to the image processing time of the image data. After determining ΔX, the position of the fluorescence detection area is adjusted accordingly. During signal acquisition, a droplet flows on the microfluidic chip, sequentially passing through the image detection area and the fluorescence detection area. Image data is acquired within the image detection area, while in the fluorescence detection area, the droplet excites and generates a fluorescence signal, which is then acquired. By controlling the positional offset of the fluorescence detection area relative to the image detection area, image data acquisition and fluorescence signal acquisition are synchronized, ensuring that the image data and fluorescence signal correspond to the same droplet.

[0021] 2. The microfluidic cell sorting instrument provided by this invention allows droplets to flow on a microfluidic chip, acquiring image data and fluorescence signals. Before acquisition, the position of the fluorescence detection area is adjusted, and an adjustable excitation spot module projects an excitation spot within the fluorescence detection area. During acquisition, the camera captures images of the droplets within the image detection area, while the PMT acquires fluorescence signals within the fluorescence detection area. The control unit controls the camera and PMT to synchronously acquire image data and fluorescence signals.

[0022] Compared to traditional microfluidic cell sorters, this system overcomes the mismatch between droplet velocity, camera frame rate, and PMT acquisition rate by aligning image and fluorescence acquisition in time. This reduces the probability of mismatch between droplet data, image data, and fluorescence signals, thereby improving the accuracy and reliability of the microfluidic cell sorter. Simultaneously, the adjustable excitation spot module adapts to different camera frame rates and processing delays, enhancing the device's versatility.

[0023] 3. The storage medium provided by the present invention enables the synchronous acquisition of image data and fluorescence data by executing the signal synchronous acquisition method described above when the computer program is run by the processor. Attached Figure Description

[0024] Figure 1 This is a flowchart of a signal synchronization acquisition method according to a specific embodiment of the present invention.

[0025] Figure 2 This is a schematic diagram of the channels of a microfluidic chip according to a specific embodiment of the present invention.

[0026] Figure 3 This is a structural block diagram of a microfluidic cell sorter according to a specific embodiment of the present invention.

[0027] Marked in the image: 1. Droplet generation area; 2. Transport channel; 3. Image detection area; 4. Fluorescence detection area; 5. Excitation spot; 6. Camera; 7. Adjustable excitation spot module; 8. PMT; 9. Control unit. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. The described embodiments are some, but not all, of the embodiments of the present invention.

[0029] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0030] Example 1: The signal synchronous acquisition method of the present invention can ensure that the fluorescence signal acquisition window and the image frame capture window coincide in droplet microfluidics technology, and the synchronous calibration operation of signal acquisition is convenient.

[0031] For the signal synchronization acquisition method of this invention, please refer to [link / reference]. Figure 1 and Figure 2 This includes the following steps: The microfluidic chip is equipped with an image detection area 3 for image data acquisition and a fluorescence detection area 4 for excitation and acquisition of fluorescence signals. Specifically, the image detection area 3 and the fluorescence detection area 4 are distributed along the flow direction of the droplet within the channel of the microfluidic chip. An excitation spot 5 is projected into the fluorescence detection area 4, which excites the droplet to generate a fluorescence signal. The droplet completes the excitation and acquisition of the fluorescence signal within the fluorescence detection area 4. 101. Obtain the positional offset ΔX of the fluorescence detection area 4 relative to the image detection area 3 at a preset flow rate. The positional offset ΔX is related to the image processing time of the image data. Specifically, the formula for calculating the positional offset ΔX is: ΔX =Vdroplet Tproc Where Vdroplet is the droplet velocity, Tproc is the image processing time, and the image processing time Tproc is the time required from image acquisition to image processing completion. Specifically, it can be calculated by subtracting the camera 6 trigger timestamp from the image processing completion timestamp.

[0032] During droplet detection, the droplet velocity Vdroplet is fixed. When the droplet velocity Vdroplet needs to be adjusted, the position offset ΔX needs to be redefined, and then the position of the fluorescence detection area 4 is adjusted according to the position offset ΔX. Specifically, the droplet velocity Vdroplet is obtained by processing the image data of the droplets within the image detection area. The droplets flow into the image detection area 3, and the camera 6 acquires the image data of the droplets within the image detection area 3. The specific formula for calculating the droplet velocity Vdroplet is as follows:

[0033] Where S represents the relative displacement of the droplet in the first frame image and the droplet in the second frame image acquired from the image detection area, and Tcam is the sampling period for each frame image. Specifically, the formula for calculating the sampling period Tcam for each frame image is:

[0034] Where F is the frame rate, the camera 6 acquires image data of the droplets in the image detection area 3 according to the frame rate F, obtains the image frame sequence, and compares the positional difference between the droplets in the first frame image and the droplets in the second frame image through the image frame sequence, or determines the distance S according to the reference mark of known size.

[0035] 102. Adjust the position of the fluorescence detection area 4 so that the distance between the fluorescence detection area 4 and the image detection area 3 is the position offset ΔX; 103. The droplet flows through the image detection area 3 and the fluorescence detection area 4 to simultaneously acquire image data and fluorescence signals of the droplet at a preset flow rate.

[0036] Specifically, the droplet flows within the channel of the microfluidic chip and passes sequentially through the image detection area 3 and the fluorescence detection area 4. When the droplet flows into the image detection area 3, it captures an image. When it flows into the fluorescence detection area 4, it generates a fluorescence signal, which is then collected in the fluorescence detection area 4.

[0037] Because the image data processing time for the droplet is longer than the fluorescence signal acquisition time, there is a delay. Offsetting the position of the fluorescence detection area 4 along the droplet's flow direction by a certain distance allows the fluorescence signal acquisition and processing to be completed within the droplet's image processing time Tproc. The image data sampling time window coincides with the fluorescence signal sampling time window, ensuring synchronous data acquisition. Compared to the traditional method of calibration using a single hardware clock, this method helps ensure that the image data and fluorescence data correspond to the same droplet, reducing the risk of data mismatch and improving analytical accuracy. Furthermore, synchronous data acquisition can be achieved with only a single adjustment of the fluorescence detection area 4, simplifying the calibration process.

[0038] Example 2: The difference from Example 1 is that the frame rate of camera 6 in this example is F=100fps, that is, 10ms / frame. According to the calculation formula of the sampling period Tcam of each frame of camera 6, the sampling period Tcam of each frame of camera 6 is 10ms. A droplet is generated on the microfluidic chip. The position offset ΔX of the fluorescence detection area 4 is determined by measuring the droplet, and the fluorescence detection area 4 is calibrated and adjusted.

[0039] When the droplet flows through the image detection area 3, the image data of the droplet is collected. Based on the comparison results of the first frame image and the second frame image of the camera 6, the relative displacement S=0.06mm is obtained. Then, the droplet velocity Vdroplet is calculated according to the formula for calculating the droplet velocity Vdroplet. The droplet velocity Vdroplet=6mm / s. Determine the image processing delay time, i.e., the image processing time Tproc. t =8ms; According to the formula for calculating the position offset ΔX, the offset ΔX = 6mm / s × 0.08s = 48µm; The position of the fluorescence detection area 4 is adjusted according to the position offset ΔX. The position of the fluorescence detection area 4 is 48µm behind the image detection area 3. Specifically, after determining the position of the fluorescence detection area 4, an excitation spot 5 can be projected into the fluorescence detection area 4 using a focusing lens and a dichroic mirror. The diameter of the excitation spot 5 is adjusted to 200µm. The flow rate of the liquid phase inside the microfluidic chip is fixed, and droplets are generated again on the microfluidic chip. The liquid phase drives the droplets to flow through the image detection area 3 and the fluorescence detection area 4. The droplets realize the synchronous acquisition of image data and fluorescence signals, and the droplets are matched with the image data and fluorescence signals. When it is necessary to adjust the liquid flow rate, repeat the above operation to obtain the corresponding position offset ΔX. The fluorescence detection area 4 and the excitation spot 5 are translated and adjusted in position within the microfluidic chip according to the offset ΔX.

[0040] Example 3: Based on the above embodiments, this embodiment provides a microfluidic cell sorter. By adjusting the position of the excitation spot 5, the image acquisition and fluorescence acquisition are aligned in time, which helps to improve the resolution and analysis accuracy of the microfluidic cell sorter.

[0041] The microfluidic cell sorting instrument of the present invention is used to simultaneously acquire image data and fluorescence signals of droplets on a microfluidic chip. The microfluidic chip is provided with an image detection area 3 and a fluorescence detection area 4. Specifically, the microfluidic chip includes channels for droplet flow, with the image detection area 3 and fluorescence detection area 4 located within the channels. A droplet generation area 1 and a transport channel 2 are provided within the channels. Droplets are generated in the droplet generation area 1 and flow along the transport channel to the image detection area 3 and fluorescence detection area 4. Specifically, multiple channels are provided, and multiple channels can simultaneously perform droplet analysis and detection.

[0042] The microfluidic cell sorting system includes: An adjustable excitation spot module 7 projects an excitation spot 5 into the fluorescence detection area 4 and adjusts the projection position of the excitation spot 5 according to the positional offset of the fluorescence detection area 4. In some specific embodiments, the adjustable excitation spot module 7 includes an excitation generator and a spatial light modulator. The laser generator emits the excitation spot 5, and the spatial light modulator quickly adjusts the position of the excitation spot 5 so that the excitation light 5 is projected into the fluorescence detection area 4.

[0043] In other specific embodiments, the adjustable excitation spot module 7 includes a rotatable focusing mirror and a driving source. The driving source is connected to the rotatable focusing mirror to drive the rotatable focusing mirror to move within the channel along the flow direction of the droplet, thereby adjusting the position of the excitation spot 5. The driving source is preferably, but not limited to, a micro motor or a stepper motor. The excitation spot 5 is finely adjusted by the adjustable excitation spot module 7, and its position in the channel can be accurately positioned within ±10µm. In other specific embodiments, the excitation spot 5 is adjusted manually by adjusting the laser generator.

[0044] Furthermore, the adjustable excitation spot module 7 can simultaneously project multiple wavelengths of excitation spot 5 in the same channel, realizing the synchronous acquisition of fluorescence signals of different wavelengths.

[0045] PMT8 collects the fluorescence signal of the droplet in the fluorescence detection area 4. After the excitation spot 5 excites the droplet to generate a fluorescence signal, PMT8 collects the fluorescence signal. While the camera 6 captures each frame of the image, PMT8 collects the fluorescence signal corresponding to the position of the excitation spot 5, so that the image data and fluorescence signal of the same droplet are synchronized.

[0046] Camera 6 and PMT8 are electrically connected to control unit 9. Control unit 9 executes the above-mentioned signal synchronization acquisition method, and controls the start of camera 6 and PMT8 to synchronize the acquisition of droplet image data and fluorescence signal.

[0047] Specifically, the control unit 9 may include a processor and a memory. These components communicate via one or more buses. Those skilled in the art will understand that the structure of the microfluidic cell sorter shown in the figure does not constitute a limitation of this application. It may be a bus-shaped structure or a star-shaped structure, and may include more or fewer components than shown, or combine certain components, or have different component arrangements.

[0048] The processor is the control center of the microfluidic cell sorter. It connects to various parts of the electronic positioning device via various interfaces and lines. By running or executing software programs and / or modules stored in the memory, and by calling data stored in the memory, it performs various functions of the electronic positioning device and / or processes data. The processor can be composed of integrated circuits (ICs), such as a single packaged IC or multiple packaged ICs with the same or different functions connected together. For example, the processor may consist only of a Central Processing Unit (CPU). In the embodiments of this application, the CPU may have a single processing core or include multiple processing cores.

[0049] The memory can be used to store the processor's execution instructions. The memory can be implemented by any type of volatile or non-volatile storage location device or a combination thereof, such as static random access memory (SRAM), electrically erasable programmable read-only memory (EEPROM), erasable programmable read-only memory (EPROM), programmable read-only memory (PROM), read-only memory (ROM), magnetic storage, flash memory, magnetic disk or optical disk.

[0050] When the execution instructions in the memory are executed by the processor, the control unit is able to perform some or all of the steps in the above-described signal synchronization acquisition method embodiment. For example, when adjusting the position of the fluorescence detection area 4, the processor processes the image data acquired by the camera 6 to obtain the droplet velocity Vdroplet, obtains the image processing time Tproc, and calculates the offset ΔX based on the droplet velocity Vdroplet. Based on the position offset ΔX, the processor drives the PMT8 and the adjustable excitation spot module 7 to move a certain distance relative to the image detection area 3 to adjust the positions of the fluorescence detection area 4 and the excitation spot 5. The processor then controls the camera 6, the laser generator, and the PMT8 to start, achieving synchronous sampling of the image and fluorescence.

[0051] In some specific embodiments, a feedback subsystem is included. This subsystem employs a closed-loop control algorithm. After the camera 6 acquires an image of the droplet, it measures the droplet velocity Vdroplet in real time and feeds it back to the control unit 9 to dynamically adjust the position offset ΔX. The fluorescence detection area 4 and the excitation spot 5 are optimized in real time to meet the synchronization requirements of multi-channel, multi-wavelength, and other operating modes.

[0052] Specifically, it also includes a data processing platform that can correlate synchronized images with fluorescence signals by droplet and provide a real-time visualization and multi-parameter cell analysis interface, applicable to large-scale cell screening and clinical applications. It is understood that the methods by which the data processing platform correlates and stores data are known to those skilled in the art and are achievable; therefore, this embodiment will not elaborate further.

[0053] Example 4: This embodiment, based on the above embodiments, provides a computer storage medium that can store a program. When run by a processor, this program can execute some or all of the steps provided in the embodiments of this application, for example: The positional offset ΔX of the fluorescence detection area relative to the image detection area at a preset flow rate is obtained. The positional offset ΔX is related to the image processing time of the image data. The position of the fluorescence detection area is adjusted so that the distance between the fluorescence detection area and the image detection area is the positional offset ΔX. The droplet flows through the image detection area and the fluorescence detection area, and the image data and fluorescence signal of the droplet at the preset flow rate are collected simultaneously.

[0054] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0055] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship commonly used when the product of this invention is in use. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this invention. In addition, the terms "first," "second," "third," etc., are only used to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0056] Furthermore, terms such as "horizontal," "vertical," and "sag" do not imply that components must be absolutely horizontal or suspended, but rather that they can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal relative to "vertical," and does not mean that the structure must be completely horizontal, but can be slightly tilted.

[0057] In this invention, unless otherwise expressly specified and limited, "above or below" a first feature may include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on" the first feature includes the first feature directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the first feature includes the first feature directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0058] Although the description of the invention has been given in conjunction with the specific embodiments described above, it will be apparent to those skilled in the art that many substitutions, modifications, and variations can be made based on the foregoing. Therefore, all such substitutions, modifications, and variations are included within the spirit and scope of the appended claims.

Claims

1. A method for synchronous signal acquisition, characterized in that, Includes the following steps: The microfluidic chip has an image detection area for acquiring image data and a fluorescence detection area for exciting and acquiring fluorescence signals; The positional offset ΔX of the fluorescence detection area relative to the image detection area at a preset droplet flow rate is obtained, and the positional offset ΔX is related to the image processing time of the image data; Adjust the position of the fluorescence detection area so that the distance between the fluorescence detection area and the image detection area is the position offset ΔX; The droplet flows through the image detection area and the fluorescence detection area, and image data and fluorescence signals of the droplet at a preset droplet flow rate are collected simultaneously.

2. The signal synchronization acquisition method according to claim 1, characterized in that, The formula for calculating the offset ΔX is: ΔX =Vdroplet Tproc Where Vdroplet is the droplet flow rate and Tproc is the image processing time.

3. The signal synchronization acquisition method according to claim 2, characterized in that, The droplet velocity Vdroplet is obtained by processing image data of droplets within the image detection area.

4. The signal synchronization acquisition method according to claim 3, characterized in that, The formula for calculating the droplet velocity Vdroplet is: Where S is the relative displacement of the droplet in the first frame image and the droplet in the second frame image acquired from the image detection area, and Tcam is the sampling period of each frame image.

5. The signal synchronization acquisition method according to claim 4, characterized in that, The formula for calculating the sampling period Tcam of each frame image is: Where F is the frame rate, and image data of droplets in the image detection area are acquired based on the frame rate F.

6. A microfluidic cell sorting device, characterized in that, The microfluidic cell sorting instrument is used for synchronous acquisition of image data and fluorescence signals of droplets on a microfluidic chip. The microfluidic chip is provided with an image detection area and a fluorescence detection area. The microfluidic cell sorting instrument includes: A camera is used to acquire images of droplets flowing through the image detection area; An adjustable excitation spot module is used to project an excitation spot within the fluorescence detection area and adjust the projection position of the excitation spot. PMT, used to acquire the fluorescence signal of the droplets within the fluorescence detection area; and The control unit is electrically connected to the camera and the PMT respectively. The control unit performs the signal synchronization acquisition method as described in any one of claims 1-5 so that the image data of the droplet and the fluorescence signal are acquired synchronously.

7. The microfluidic cell sorting device according to claim 6, characterized in that, The adjustable excitation spot module includes a spatial light modulator, which adjusts the position of the excitation spot according to the offset ΔX.

8. The microfluidic cell sorting device according to claim 6, characterized in that, It includes a feedback subsystem that measures the droplet flow rate Vdroplet in real time after the camera acquires an image of the droplet in order to dynamically adjust the offset ΔX of the excitation spot.

9. The microfluidic cell sorting device according to claim 6, characterized in that, The microfluidic chip includes a channel for droplet flow, with the image detection area and fluorescence detection area located within the channel. The channel is provided with a droplet generation area and a transport channel. The droplets are generated in the droplet generation area and flow along the transport channel to the image detection area and the fluorescence detection area.

10. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a computer program; The computer program is executed by the processor to perform the signal synchronization acquisition method as described in any one of claims 1-5.

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