Particle printing method, system and device based on fluorescent field and bright field time sequence imaging

Abstract

The invention discloses a particle printing method, system and device based on fluorescent field and bright field time sequence imaging, and the method comprises the steps: monitoring a micro-fluidic chip sorting region under the condition of continuous illumination of a multi-color fluorescent field, and collecting a fluorescent image of the micro-fluidic chip sorting region in real time; when the target fluorescent particles are recognized in the fluorescent image, bright field illumination is automatically triggered, and a bright field image is collected; carrying out fusion analysis processing on the fluorescence image and the bright field image, and extracting multi-dimensional feature parameters; judging whether the multi-dimensional characteristic parameters meet printing conditions or not according to a preset multi-modal judgment rule; when the multi-dimensional characteristic parameters meet printing conditions, a printing instruction is sent; and in response to the printing instruction, controlling the pressure and motion control unit to drive the printing channel of the micro-fluidic chip to print the target fluorescent particles to the specified hole site of the microwell plate. According to the particle printing technology combining the advantages of fluorescence imaging and bright field imaging, comprehensive feature analysis and high-precision sorting of particles are achieved.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and microbiology research technology, and in particular to a method, system and apparatus for microparticle printing based on fluorescence field and bright field time-series imaging. Background Technology

[0002] Fluorescent sample sorting and printing technology has significant application value in fields such as biomedicine and microbiology research. However, current fluorescent particle printing methods typically rely solely on the fluorescence signal generated by laser excitation as the sorting and printing criterion, failing to identify and exclude non-fluorescent particles, and unable to simultaneously identify and distinguish between fluorescent and non-fluorescent particles. This single signal discrimination mechanism makes it difficult to achieve accurate screening and printing of mixed samples, severely limiting its application in complex biological samples.

[0003] Furthermore, current fluorescent microparticle printing methods only acquire fluorescence signals and cannot provide bright-field images. This makes it difficult to achieve unified imaging and synchronous analysis of bright-field and multicolor fluorescence. Consequently, it is impossible to analyze the morphology and target features of particles or cells. The lack of morphology and target feature information of fluorescent microparticles makes it difficult to achieve multidimensional identification and quality judgment of target fluorescent microparticles. It is also impossible to exclude non-target components such as fragments and aggregates, making it difficult to perform accurate identification and quality control.

[0004] In addition, existing methods for detecting fluorescent particles acquire multicolor fluorescence signals by configuring multiple photomultiplier tubes or photodiodes. However, each detection channel only outputs light intensity values, not spatial imaging information. Furthermore, the signals between different channels are independent, making spatial alignment and morphology analysis impossible. They also cannot achieve rapid switching and joint acquisition of fluorescence and bright field illumination in high-speed flowing samples. Therefore, they cannot simultaneously utilize fluorescence information and particle morphology characteristics for comprehensive judgment. Currently, these methods have significant shortcomings in the sorting of multicolor labeled samples, morphology verification, and identification of complex samples. Summary of the Invention

[0005] This invention provides a microparticle printing method, system, and apparatus based on fluorescence field and bright field time-series imaging, aiming to solve at least one of the technical problems existing in the prior art.

[0006] The technical solution of this invention is a microparticle printing method based on fluorescence field and bright field temporal imaging, which includes: Under continuous illumination of a multicolor fluorescence field, the sorting area of ​​the microfluidic chip is monitored, and fluorescence images of the sorting area of ​​the microfluidic chip are acquired in real time. When the target fluorescent particles are identified in the fluorescence image, bright-field illumination is automatically triggered, and a bright-field image is acquired. The fluorescence image and the bright field image are fused and analyzed, and multidimensional feature parameters are extracted. The multidimensional feature parameters are determined according to the preset multimodal judgment rules to see if they meet the printing conditions. When the multidimensional feature parameters meet the printing conditions, a printing command is sent. In response to the printing command, the pressure and motion control unit drives the printing channel of the microfluidic chip to print the target fluorescent microparticles onto the designated wells of the microplate.

[0007] According to some embodiments of the present invention, the fusion analysis of the fluorescence image and the bright field image, and the extraction of multidimensional feature parameters, includes: The fluorescence image is analyzed and processed by the image processing unit to obtain the target feature parameters of the fluorescence image, including the fluorescence intensity and fluorescence color of the target fluorescent particles; The bright-field image is analyzed and processed by the image processing unit to obtain the target morphology parameters of the bright-field image, including the size of the target fluorescent particles and the roundness of the target fluorescent particles.

[0008] According to some embodiments of the present invention, determining whether the multidimensional feature parameters meet the printing conditions based on preset multimodal judgment rules includes: When the fluorescence intensity of the target fluorescent particles is within a preset range and the fluorescence color of the target fluorescent particles is consistent with the set color, it is determined that the target feature parameters of the fluorescence image satisfy the preset multimodal judgment rule; When the size of the target fluorescent microparticle is within a preset range and the roundness of the target fluorescent microparticle is within a preset range, it is determined that the target morphology parameters of the bright field image satisfy the preset multimodal judgment rule; When the target feature parameters of the fluorescence image and the target morphology parameters of the bright field image simultaneously satisfy the preset multimodal judgment rule, it is determined whether the multidimensional feature parameters meet the printing conditions.

[0009] According to some embodiments of the present invention, it further includes: The fluorescence image is uploaded to the image processing unit; The image processing unit performs real-time processing on the fluorescence image based on a recognition algorithm to identify potential targets; Determine whether the potential target meets the preset verification conditions; When the potential target meets the preset verification conditions, the potential target is identified as the target fluorescent microparticle.

[0010] According to some embodiments of the present invention, the multicolor fluorescence field adopts a multicolor LED array and filter wheel scheme, and intelligent switching of multicolor fluorescence is achieved through timing control and spectral matching; The multi-color LED array includes a three-color LED array composed of ultraviolet, blue, and green LEDs. The filter wheel is driven by a stepper motor and completes the filter switching within a preset time to uniformly collect fluorescence signals under multi-wavelength excitation.

[0011] According to some embodiments of the present invention, it further includes: If the multidimensional feature parameters do not meet the printing conditions, immediately turn off the bright field illumination, restore the multicolor fluorescence field monitoring state, and wait for the next potential target to appear.

[0012] According to some embodiments of the present invention, it further includes: After determining whether the multidimensional feature parameters meet the printing conditions, it is determined whether the number of target fluorescent particles printed meets the set threshold. If the number of printed target fluorescent particles does not meet the set threshold, the bright field illumination is turned off, and the system returns to the multicolor fluorescence field monitoring state, waiting for the next potential target to appear. If the number of target fluorescent particles printed meets the set threshold, the printing operation ends.

[0013] According to some embodiments of the present invention, it further includes: Once the printing operation is complete, turn off the bright field illumination and return to the multicolor fluorescence field monitoring state, waiting for the next potential target to appear.

[0014] The technical solution of the present invention also includes a microparticle printing system based on fluorescence field and bright field temporal imaging, used to perform the microparticle printing method based on fluorescence field and bright field temporal imaging as described in the above embodiments, including: A microfluidic chip includes a sorting area and a printing channel. The sorting area is used to hold a microparticle sample to be sorted, and the printing channel is used to provide a flow path for the target fluorescent microparticles to perform the printing operation. An optical imaging unit includes a multi-wavelength LED light source and a camera. The multi-wavelength LED light source is used to provide multi-color fluorescence field illumination. The camera is used to monitor the sorting area of ​​the microfluidic chip under continuous multi-color fluorescence field illumination and to acquire fluorescence images of the sorting area of ​​the microfluidic chip in real time. When a target fluorescent particle is identified in the fluorescence image, bright field illumination is automatically triggered and a bright field image is acquired. The image processing unit is used to perform fusion analysis on the fluorescence image and the bright field image, extract multi-dimensional feature parameters, determine whether the multi-dimensional feature parameters meet the printing conditions according to the preset multi-modal judgment rules, and send a printing command when the multi-dimensional feature parameters meet the printing conditions. A pressure and motion control unit is used to drive the printing channel of the microfluidic chip to print the target fluorescent microparticles to the designated wells of the microplate in response to the printing command.

[0015] The present invention also relates to a computer device, including a memory and a processor, wherein the processor performs the above-described method when executing a computer program stored in the memory.

[0016] The present invention also relates to a computer-readable storage medium storing computer program instructions thereon, which, when executed by a processor, implement the above-described method.

[0017] The microparticle printing method, system, and apparatus based on fluorescence field and bright field temporal imaging provided in this invention have at least one of the following advantages or beneficial effects: By real-time monitoring of the sorting area of ​​the microfluidic chip, when a target fluorescent microparticle is identified, bright field illumination is automatically triggered and the discrimination stage begins. The bright field illumination remains on throughout the discrimination process of the target fluorescent microparticle, tracking the position of the target fluorescent microparticle in the microfluidic chip's selection area in real time. After the discrimination is completed, the illumination is immediately turned off, and the system returns to the fluorescence monitoring state, waiting for the next trigger event. This achieves on-demand switching and efficient management of the light field, thereby achieving a balance between recognition efficiency and target activity.

[0018] By establishing multimodal judgment rules, the printing command is triggered only when all characteristic parameters of the target fluorescent microparticles meet the judgment benchmark thresholds, significantly improving the accuracy and reliability of sorting. Utilizing the synergistic effect of the microfluidic chip printing channel and the pressure and motion control unit, the pressure in the printing area and the movement of the microplate are synchronously adjusted to ensure that each qualified target fluorescent microparticle is accurately assigned to the designated well in the microplate, achieving fully automated operation from target fluorescent microparticle identification to positioning and printing.

[0019] Furthermore, additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0020] Figure 1 This is a flowchart of the overall process of the microparticle printing method based on fluorescence field and bright field temporal imaging provided in the embodiments of the present invention; Figure 2 This is a structural diagram of the microfluidic chip provided in the embodiments of the present invention; Figure 3 This is a schematic diagram of the flow trajectory of the target fluorescent particles provided in an embodiment of the present invention; Figure 4 This is a detailed flowchart of step S300 in the microparticle printing method based on fluorescence field and bright field temporal imaging provided in the embodiments of the present invention; Figure 5 This is a detailed flowchart of step S400 in the microparticle printing method based on fluorescence field and bright field temporal imaging provided in the embodiments of the present invention; Figure 6 This is a detailed flowchart of the first method for microparticle printing based on fluorescence field and bright field temporal imaging provided in the embodiments of the present invention; Figure 7 This is a schematic diagram of cell recognition verification based on fluorescence field and bright field timing switching provided in an embodiment of the present invention; Figure 8 This is a schematic diagram of the imaging effect of microparticles under original fluorescent illumination provided in an embodiment of the present invention; Figure 9 This is a schematic diagram of the imaging effect of fluorescently illuminated microparticles provided in an embodiment of the present invention; Figure 10 This is a second detailed flowchart of the microparticle printing method based on fluorescence field and bright field temporal imaging provided in the embodiments of the present invention; Figure 11 This is a schematic diagram of the microparticle printing system based on fluorescence field and bright field temporal imaging provided in an embodiment of the present invention. Detailed Implementation

[0021] The following will provide a clear and complete description of the concept, specific structure, and technical effects of the present invention in conjunction with the embodiments and accompanying drawings, so as to fully understand the purpose, solution, and effects of the present invention.

[0022] It should be noted that, unless otherwise specified, when a feature is referred to as "fixed" or "connected" to another feature, it can be directly fixed or connected to the other feature, or indirectly fixed or connected to the other feature. The singular forms "a," "described," and "the" used herein are also intended to include the plural forms, unless the context clearly indicates otherwise. Furthermore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used in this specification is for the purpose of describing particular embodiments only and not for limiting the invention. The term "and / or" as used herein includes any combination of one or more of the associated listed items.

[0023] It should be understood that although the terms first, second, third, etc., may be used to describe various elements in this disclosure, these elements should not be limited to these terms. These terms are used only to distinguish elements of the same type from one another. For example, a first element may also be referred to as a second element without departing from the scope of this disclosure, and similarly, a second element may also be referred to as a first element. Any and all instances or exemplary language (“e.g.,” “such as,” etc.) provided herein are intended only to better illustrate embodiments of the invention and, unless otherwise required, do not impose a limitation on the scope of the invention.

[0024] This invention provides a microparticle printing method, system, and device based on fluorescence field and bright field time-series imaging. This microparticle printing technology combines the advantages of fluorescence imaging and bright field imaging to achieve comprehensive feature analysis and high-precision sorting of target fluorescent microparticles, meeting the high requirements for microparticle sorting and printing in the fields of biomedicine and microbiology research.

[0025] Please refer to the following. Figures 1 to 11 The present invention provides a further description of the microparticle printing method, system, and apparatus based on fluorescence field and bright field temporal imaging.

[0026] Reference Figure 1 As shown, Figure 1 This is a general flowchart of the microparticle printing method based on fluorescence field and bright field temporal imaging provided in this embodiment of the invention. The microparticle printing method based on fluorescence field and bright field temporal imaging includes, but is not limited to, steps S100 to S600. Specifically, S100: Under continuous illumination of a multicolor fluorescence field, the sorting area of ​​the microfluidic chip is monitored, and fluorescence images of the sorting area of ​​the microfluidic chip are acquired in real time. S200: When target fluorescent particles are identified in a fluorescence image, bright-field illumination is automatically triggered and a bright-field image is acquired; S300: Performs fusion analysis on fluorescence and bright-field images and extracts multidimensional feature parameters; S400: Determines whether the multi-dimensional feature parameters meet the printing conditions according to the preset multi-modal judgment rules; S500: When the multidimensional feature parameters meet the printing conditions, send a printing command; S600: In response to a printing command, the pressure and motion control unit drives the printing channel of the microfluidic chip to print target fluorescent microparticles onto the designated wells of the microplate.

[0027] In some embodiments of the present invention, the microparticle printing method based on fluorescence field and bright field time-series imaging includes: continuously illuminating the sorting area of ​​a microfluidic chip using multiple light sources of different wavelengths. This multicolor illumination can excite different types of fluorescent microparticles to emit fluorescence at specific wavelengths, thereby enabling simultaneous monitoring of multiple fluorescent microparticles. Fluorescence images of the sorting area are acquired in real time using a high-sensitivity imaging device (such as a fluorescence microscope or a high-resolution camera).

[0028] Leveraging the parallel computing capabilities of the graphics processing unit (GPU), fluorescence images are processed in real-time using image processing algorithms (such as dynamic target recognition and noise reduction) to identify fluorescent particles that meet preset conditions, including features such as fluorescence intensity and color. When a target fluorescent particle is identified in the fluorescence image, the system automatically switches to and remains in bright-field illumination mode, simultaneously acquiring and analyzing bright-field images. Bright-field illumination refers to illuminating the microfluidic chip with ordinary white light and acquiring bright-field images. These images provide information such as particle morphology and size, complementing the fluorescence images.

[0029] By employing a high-resolution camera and multiple light sources of different wavelengths, embodiments of the present invention can achieve synchronous or temporal acquisition of multicolor fluorescence signals (target fluorescent particles) and bright-field images under a single optical path. Bright-field imaging is triggered immediately upon the appearance of the target fluorescent particles, and the target fluorescent particles are verified by combining the fluorescence image and the bright-field image. This overcomes the limitations of existing laser sorting systems that rely on multiple PMT detectors, lack bright-field information, and have spatial alignment capabilities, thus achieving efficient fusion sorting of multicolor fluorescence and bright-field information.

[0030] By leveraging the parallel computing capabilities of the graphics processing unit, fluorescence images and bright-field images are fused. Fusion methods can include simple overlay, feature-based fusion algorithms (such as wavelet transform fusion), etc., to quickly extract multidimensional feature parameters of the target fluorescent particles in the fused image. These multidimensional feature parameters may include fluorescence intensity, fluorescence color, particle size, shape, and position. These multidimensional feature parameters are used for subsequent judgment and analysis.

[0031] The extracted multidimensional feature parameters are comprehensively judged according to the preset multimodal judgment rules. The preset multimodal judgment rules set multiple judgment benchmark thresholds for feature parameters. The extracted multidimensional feature parameters are judged against the set judgment benchmark thresholds to determine whether the target fluorescent microparticles meet the printing conditions. If the multidimensional feature parameters meet the preset printing conditions, the printing command is triggered.

[0032] Responding to the printing command, the pressure and motion control unit precisely drives the printing channel of the microfluidic chip, accurately printing a single target fluorescent particle into the designated well of the microplate, achieving efficient and reliable sorting of "one target fluorescent particle per well".

[0033] Understandably, the microparticle printing method based on fluorescence and bright-field temporal imaging monitors the sorting area of ​​the microfluidic chip in real time. When a target fluorescent microparticle is detected, bright-field illumination is automatically triggered, and the process enters the discrimination phase. The bright-field illumination remains on throughout the discrimination process, and is immediately turned off after the discrimination is completed, then returns to the fluorescence monitoring state, awaiting the next trigger event. This printing method achieves on-demand switching and efficient management of the light field, thus striking a balance between recognition efficiency and target activity. By establishing multimodal judgment rules, the printing command is triggered only when all characteristic parameters of the target fluorescent microparticle meet the judgment benchmark threshold, significantly improving the accuracy and reliability of sorting.

[0034] Based on the precise flow channel and functional area design of microfluidic chips (such as...) Figure 2 As shown), by integrating fluorescence images, bright-field images, and printing instructions, the position of target fluorescent particles in the selected area of ​​the microfluidic chip can be tracked in real time (e.g., ...). Figure 3 As shown, by utilizing the synergistic effect of the microfluidic chip printing channel and the pressure and motion control unit, the pressure in the printing area and the displacement of the collection platform containing the microplate are synchronously adjusted to achieve fully automated operation of the target fluorescent microparticle from identification to positioning and printing. This closed-loop control system ensures that each qualified target fluorescent microparticle is accurately assigned to the designated well in the microplate, achieving high-precision sorting at the single-target level.

[0035] Reference Figure 4 As shown, Figure 4 This is a detailed flowchart of step S300 in the microparticle printing method based on fluorescence field and bright field temporal imaging provided in this embodiment of the invention. Step S300 includes, but is not limited to, steps S310 to S320. Specifically, S310: The fluorescence image is analyzed and processed by the image processing unit to obtain the target feature parameters of the fluorescence image. The target feature parameters include the fluorescence intensity and fluorescence color of the target fluorescent particles. S320: The image processing unit analyzes and processes the bright-field image to obtain the target morphology parameters of the bright-field image, including the size of the target fluorescent particles and the roundness of the target fluorescent particles.

[0036] In some embodiments of the present invention, the fluorescence image and the bright field image need to be preprocessed before extracting the multidimensional feature parameters of the fluorescence image and the bright field image in order to improve the image quality and the accuracy of the analysis.

[0037] The preprocessing steps for fluorescence images include: The fluorescence image was binarized. A dynamic target recognition algorithm is used to capture targets in fluorescence images, which facilitates subsequent processing. Morphological algorithms were used to further denoise the fluorescence images, resulting in high-quality fluorescence images. The location of the target fluorescent microparticles was confirmed using a contour-finding algorithm.

[0038] Subsequently, the image processing unit performs intensity statistics on the target region of the fluorescence image, calculating the average fluorescence intensity, maximum fluorescence intensity, or total fluorescence intensity, and analyzes the fluorescence image to obtain the fluorescence intensity information of the target fluorescent particles. It is understandable that fluorescence intensity can reflect the concentration, activity, or labeling efficiency of fluorescent particles. For example, in biological experiments, fluorescence intensity can be used to quantitatively analyze cell activity or protein expression levels. It is also understandable that fluorescence color can be used to distinguish different types of fluorescent particles or labels. In multi-labeling experiments, different colors of fluorescence can be used simultaneously to label different biomolecules or cell types.

[0039] The preprocessing steps for brightfield images include: Gaussian blurring was applied to the bright field image to initially eliminate contour spikes; The dynamic target recognition algorithm is used to remove the background from the bright field image, leaving only the dynamic target. At the same time, the bright field image is normalized by the dynamic target recognition algorithm. Morphological algorithms are used to further denoise the bright field image; A contour-finding algorithm is used to determine the location of the target fluorescent microparticles; The location of the target fluorescent particles is captured in a screenshot, and the relevant parameters of the target fluorescent particles in the screenshot are calculated.

[0040] Understandably, the size of a target fluorescent particle can reflect its physical properties. For example, in biological experiments, cell size can be used to distinguish different types of cells or assess the growth status of cells.

[0041] In some embodiments of the present invention, the target morphology parameters also include the roundness of the target fluorescent microparticles. The roundness of the target fluorescent microparticles can reflect whether the shape of the cell meets the requirements. When the roundness of the target fluorescent microparticles is within a preset range, it is confirmed that the target fluorescent microparticles meet the printing conditions, and then the target fluorescent microparticles are selected for subsequent experiments to improve the accuracy and success rate of the experiments.

[0042] Understandably, particle size can be used to assess morphological changes in target fluorescent particles. In biological experiments, this parameter can be used to analyze morphological changes in cells, such as cell spreading and contraction.

[0043] By analyzing and processing fluorescence and bright-field images, and combining their feature parameters, multidimensional feature parameters such as fluorescence intensity, fluorescence color, particle size, and particle roundness can be extracted. This allows for a more comprehensive description of the characteristics of fluorescent particles. These multidimensional feature parameters are of great significance in multimodal analysis, providing important basis for subsequent multimodal judgment and printing operations. This multimodal analysis method has broad application prospects in biomedical research, drug screening, and biochip manufacturing, and can improve the accuracy and efficiency of experiments.

[0044] Reference Figure 5 As shown, Figure 5 This is a detailed flowchart of step S400 in the microparticle printing method based on fluorescence field and bright field temporal imaging provided in this embodiment of the invention. Step S400 includes, but is not limited to, steps S410 to S430. Specifically, S410: When the fluorescence intensity of the target fluorescent particles is within a preset range and the fluorescence color of the target fluorescent particles is consistent with the set color, determine that the target feature parameters of the fluorescence image meet the preset multimodal judgment rules. S420: When the size of the target fluorescent particles is within a preset range and the roundness of the target fluorescent particles is within a preset range, the target morphology parameters of the bright field image are determined to meet the preset multimodal judgment rules. S430: When the target feature parameters of the fluorescence image and the target morphology parameters of the bright field image simultaneously satisfy the preset multimodal judgment rules, determine whether the multidimensional feature parameters meet the printing conditions.

[0045] In some embodiments of the present invention, determining whether the multidimensional feature parameters meet the printing conditions according to preset multimodal judgment rules includes: Determining whether the fluorescence intensity and color of a target fluorescent particle meet preset multimodal judgment rules can be understood as follows: the fluorescence intensity threshold is used to filter out target fluorescent particles with fluorescence intensity that fully meets the requirements, avoiding misjudgment or inaccurate detection due to weak signals. Other modalities are only judged when the fluorescence intensity is within the preset range, improving recognition efficiency and accuracy.

[0046] Consistency in fluorescence color is used to distinguish different types of fluorescent particles or labels. For example, in multi-label experiments, different colors of fluorescence may represent different biomolecules or cell types. In multicolor fluorescence imaging, green fluorescence may label the cell membrane, while red fluorescence may label the cell nucleus. Only when the fluorescence color matches the preset color is the particle considered the target fluorescent particle.

[0047] Therefore, when the fluorescence intensity of the target fluorescent particles is within a preset range and the fluorescence color of the target fluorescent particles is consistent with the set color, the target feature parameters of the fluorescence image are determined to meet the preset multimodal judgment rules.

[0048] Determining whether the size and roundness of target fluorescent microparticles meet preset multimodal judgment rules can be understood as follows: the size range of target fluorescent microparticles is used to screen out microparticles that meet specific physical dimensions. The roundness range of target fluorescent microparticles is used to further verify whether the particle morphology meets the requirements. The roundness of target fluorescent microparticles can reflect the two-dimensional morphological characteristics of the microparticles, such as the spreading state of cells. In cell morphology analysis, a cell is considered to have a normal morphology only when both cell size and roundness are within the normal range.

[0049] Therefore, when the size of the target fluorescent particles is within a preset range and the roundness of the target fluorescent particles is within a preset range, the target morphology parameters of the bright field image are determined to meet the preset multimodal judgment rules.

[0050] Subsequently, a comprehensive judgment of multidimensional feature parameters is performed. Only when the target feature parameters (fluorescence intensity and fluorescence color) of the fluorescence image and the target morphology parameters (particle size and particle roundness) of the bright-field image simultaneously meet the preset multimodal judgment rules are the multidimensional feature parameters considered to meet the printing conditions. This comprehensive judgment method ensures that the target fluorescent particles meet the requirements in both fluorescence and morphology, thereby improving the accuracy and reliability of screening.

[0051] In one embodiment of the present invention, cell sorting and printing includes setting fluorescence conditions and bright-field conditions, and determining whether the target characteristic parameters and target morphology parameters of the target fluorescent microparticles meet the printing conditions. Specifically, Fluorescence conditions: Set the preset range of fluorescence intensity to 100 to 260 (imaging fluorescence intensity), and set the fluorescence color to green (wavelength 550nm).

[0052] Bright field conditions: Set the cell size to a preset range of 10 to 20 μm and the roundness to a range of 0.7 to 1.

[0053] Judgment process: If a cell has a fluorescence intensity of 120 (within the preset range), a fluorescence color of green (consistent with the set fluorescence color), a size of 15 μm (within the preset range), and a roundness of 0.8 (within the preset range), then the cell meets all the conditions and can be printed into the specified well of the microplate.

[0054] If a cell has a fluorescence intensity of 80 (outside the preset range), the cell will not meet the printing requirements, even if other conditions are met.

[0055] This multimodal judgment rule, by integrating the feature parameters of fluorescence and bright-field images, ensures that the target fluorescent microparticles meet preset conditions in both fluorescence signal and morphology. This method has significant application value in biomedical research, cell sorting, drug screening, and biochip manufacturing, and can significantly improve the accuracy and reliability of experiments.

[0056] This invention utilizes an image processing unit (GPU accelerator) to extract multi-dimensional features from continuously acquired fluorescence and bright-field images, including parameters such as fluorescence intensity, fluorescence color, particle size, and roundness. By establishing a comprehensive discrimination process that combines fluorescence intensity and color in the fluorescence field with bright-field morphology features and threshold judgment, intelligent identification and screening of fluorescent particles, non-fluorescent particles, and targets with abnormal shapes is achieved. This significantly improves sorting accuracy and recognition reliability, ensuring that each target fluorescent particle is precisely printed to a preset position within the microfluidic chip, achieving high-precision sorting of one target per aperture.

[0057] Reference Figure 6 As shown, Figure 6 This is a detailed flowchart of the first embodiment of the microparticle printing method based on fluorescence field and bright field temporal imaging provided by the present invention. The microparticle printing method based on fluorescence field and bright field temporal imaging also includes, but is not limited to, steps S110 to S140. Specifically, S110: Upload the fluorescence image to the image processing unit; S120: The image processing unit performs real-time processing of the fluorescence image based on the recognition algorithm to identify potential targets; S130: Determine whether the potential target meets the preset verification conditions; S140: When a potential target meets the preset verification conditions, the potential target is identified as the target fluorescent microparticle.

[0058] In some embodiments of the present invention, the microparticle printing method based on fluorescence field and bright field time-series imaging further includes: acquiring fluorescence images using a high-resolution camera; firstly, the fluorescence images are acquired using a high-resolution camera, and these fluorescence images contain fluorescence signal information of the fluorescent microparticles; then, the fluorescence images are transmitted to an image processing unit. The image processing unit is a GPU accelerator; after receiving the fluorescence images, the image processing unit stores them in memory or on a hard disk for subsequent processing and analysis.

[0059] The image processing unit uses a preset recognition algorithm to process the fluorescence image in real time, with the aim of identifying potential targets from the image. The recognition algorithm may include the following: Threshold segmentation: By setting a fluorescence intensity threshold, regions in the fluorescence image that fall within the threshold range are identified as potential targets.

[0060] Target segmentation: Using a dynamic target recognition algorithm, we find dynamic potential fluorescent particles and obtain screenshots of the fluorescent particles.

[0061] Color matching: After determining that the fluorescent microparticle meets the preset fluorescence intensity, the color of the fluorescent microparticle is determined according to the color channel to see if it matches the color requirement. If they match, it is determined to be the target fluorescent microparticle.

[0062] After identifying potential targets, it is necessary to further verify whether these targets meet the preset conditions, referring to... Figure 7 As shown, Figure 7 This is a schematic diagram illustrating cell recognition verification using fluorescence field / bright field timing switching; specifically, it determines whether the following parameters meet the preset conditions: Fluorescence intensity range: Whether the fluorescence intensity of the potential target is within the preset range.

[0063] Fluorescence color consistency: Whether the fluorescence color of the potential target is consistent with the set color.

[0064] Morphological characteristics: Whether the size, shape, roundness, etc. of the potential target meet the preset morphological requirements.

[0065] Location information: Whether a potential target appears in a specific region of the image (e.g., some experiments may only focus on particles in a specific location).

[0066] The characteristic parameters of potential targets are verified through a series of logical judgments or algorithms. Once a potential target passes all preset verification conditions, it is officially identified as a target fluorescent microparticle. This means that the microparticle meets the experimental requirements and can proceed to subsequent processing steps (such as printing and sorting). Once the target fluorescent microparticle is confirmed, the corresponding printing operation is triggered to print the target fluorescent microparticle to the designated location.

[0067] Afterwards, relevant information about the target fluorescent microparticles (such as location, characteristic parameters, etc.) is recorded for subsequent data analysis and experimental recording. The relevant information about the target fluorescent microparticles is also fed back to the user for manual intervention or further analysis.

[0068] By uploading fluorescence images to an image processing unit and processing them in real time based on a recognition algorithm, potential targets can be quickly identified. Further verification of these potential targets ensures that only qualified fluorescent microparticles are identified and proceed to subsequent processing steps. This method has significant application value in biomedical research, cell sorting, and biochip manufacturing, significantly improving experimental efficiency and accuracy.

[0069] In some embodiments of the present invention, the multicolor fluorescence field adopts a multicolor LED array and filter wheel scheme, and realizes intelligent switching of multicolor fluorescence through timing control and spectral matching; wherein, the multicolor LED array includes a three-color LED array composed of ultraviolet, blue and green, and the filter wheel is driven by a stepper motor to complete the filter switching within a preset time to uniformly collect fluorescence signals under multi-wavelength excitation.

[0070] Understandably, the multicolor fluorescence field employs a multicolor LED array, which consists of LEDs of three different wavelengths: ultraviolet, blue, and green. These three types of LEDs emit light of different wavelengths to excite different types of fluorescent markers.

[0071] The filter wheel consists of multiple filters, each corresponding to a specific wavelength range. Driven by a stepper motor, the filter wheel can switch filters within a preset time. The filter wheel is used to select excitation light of a specific wavelength, ensuring that only light of that specific wavelength can pass through and excite the fluorescent label. Simultaneously, the filter wheel can also be used to select emission light of a specific wavelength to collect specific fluorescence signals. By switching filters, the excitation and emission spectra of different fluorescent labels can be precisely matched. Users can flexibly configure filters according to experimental needs, supporting imaging of various fluorescent labels.

[0072] Timing control allows for precise control of the switching on and off of the multi-color LED array and the switching of the filter wheel. This precise control ensures that each acquired fluorescence signal corresponds to a specific wavelength of excitation light. This avoids interference between excitation light of different wavelengths, improving the accuracy and reliability of imaging. Through precise timing control and rapid switching of the filter wheel, fluorescence signals under multi-wavelength excitation can be acquired uniformly within a preset time period. This means that fluorescence signals of each wavelength can be acquired within the same time interval, thus guaranteeing data consistency and comparability.

[0073] In one embodiment, a high-sensitivity camera and a precisely optimized fluorescence optical path are employed. The fluorescence optical path utilizes a multi-color LED array and a filter wheel scheme, achieving intelligent switching of multi-color fluorescence through timing control and spectral matching. Specifically, the system integrates a three-color LED array (UV, 378nm), green (550nm), and blue (470nm), along with corresponding excitation and emission filters. The filter wheel is driven by a stepper motor, enabling filter switching within 300ms, ensuring signal uniformity and acquisition efficiency under multi-wavelength excitation. For example, using a blue LED fluorescence light source to excite K562 cells results in a green image on the camera. Bright-field illumination undergoes simultaneous optical improvements, employing an LED light source and uniformity control design to enhance imaging contrast. In camera parameter settings, systematic adjustments to parameters such as exposure time ensure that both fluorescence and bright-field channels can rapidly acquire stable images with clear details under different experimental conditions (e.g., ...). Figure 8 , Figure 9 (as shown) Figure 8 The imaging effect of particles illuminated by the original fluorescence is linear, which is not conducive to subsequent processing. Figure 9 The microparticles illuminated by fluorescence appear spherical in image, making feature discrimination easier and providing a high-quality raw data foundation for subsequent analysis. The image processing stage will be adapted to this hardware upgrade, utilizing a GPU accelerator to achieve multimodal feature fusion and real-time determination based on morphology and fluorescence.

[0074] By combining a multicolor LED array and a filter wheel, along with timing control and spectral matching techniques, intelligent switching of multicolor fluorescence can be achieved, and fluorescence signals under multi-wavelength excitation can be uniformly acquired. This approach has significant application value in fields such as biomedical imaging, cell analysis, and drug screening, and can significantly improve experimental efficiency and accuracy.

[0075] Furthermore, it should be noted that this invention uses an LED light source to provide low-power-density, uniform illumination, avoiding laser optical interference and the phototoxicity and photobleaching effects of high-power light on the samples. Simultaneously, the single-path, single-camera structure achieves miniaturization and high integration of the device, providing technical support for building a high-throughput, low-damage, multimodal single-sample sorting and printing platform.

[0076] In some embodiments of the present invention, the microparticle printing method based on fluorescence field and bright field time-series imaging further includes step S610, specifically: S610: When the multidimensional feature parameters do not meet the printing conditions, immediately turn off the bright field illumination, restore the multicolor fluorescence field monitoring state, and wait for the next potential target to appear.

[0077] By analyzing the feature parameters extracted from fluorescence and bright-field images (such as fluorescence intensity, fluorescence color, particle size, and roundness), the system determines whether these parameters meet the preset printing conditions. If any one or more feature parameters fail to meet the preset requirements, the system will determine that the current target does not meet the printing conditions. For example, if the fluorescence intensity is not within the preset range; the fluorescence color is inconsistent with the set color; or the particle size or roundness exceeds the preset range, the system will immediately turn off the bright-field imaging to prepare for the next fluorescence signal capture.

[0078] When the multidimensional feature parameters do not meet the printing conditions, the bright field illumination is immediately turned off. This operation is to save resources (such as light source lifespan and system energy consumption) and reduce unnecessary exposure time, so as to avoid unnecessary light damage to the sample.

[0079] After turning off the bright field illumination, immediately resume the multicolor fluorescence field monitoring state, continue to perform fluorescence imaging on the sorting area of ​​the microfluidic chip, resume monitoring state to continue searching for the next potential target fluorescent particles, and ensure continuous operation and efficient screening of targets that meet the requirements.

[0080] Under multicolor fluorescence field monitoring, fluorescence images are continuously acquired to search for new potential target fluorescent microparticles. The newly acquired fluorescence images are analyzed in real time by an image processing unit to identify potential targets and verify whether their characteristic parameters meet the printing conditions. This process is repeated until a target fluorescent microparticle meeting the printing conditions is found. This mechanism ensures the system's high-throughput screening capability, improves experimental efficiency, and, moreover, the highly automated process reduces manual intervention, improving the system's stability and reliability.

[0081] In some embodiments of the present invention, the microparticle printing method based on fluorescence field and bright field time-series imaging further includes step S620, specifically: S620: After the printing operation is completed, turn off the bright field illumination and return to the multicolor fluorescence field monitoring state, waiting for the next potential target to appear.

[0082] Once the multidimensional characteristic parameters of the target fluorescent microparticles meet the printing conditions, a printing command is triggered, controlling the printing channel of the microfluidic chip to print the target fluorescent microparticles onto the designated wells of the microplate. After the printing operation is completed, a confirmation signal (such as a flag indicating that the printing command has been completed) will be received, indicating that the printing task has been successfully executed.

[0083] After the printing operation is completed, immediately turn off the bright-field illumination to reduce the usage time of the light source, extend its lifespan, and reduce energy consumption. This avoids unnecessary light damage to the sample or particles in the microfluidic chip. After turning off the bright-field illumination, immediately resume the multicolor fluorescence field monitoring state and continue fluorescence imaging of the sorting area of ​​the microfluidic chip. Resuming the monitoring state is to continue searching for the next potential target fluorescent particles, preparing for the next monitoring cycle, ensuring that the system can quickly respond to the emergence of the next potential target, and ensuring that the system can continue to operate and efficiently screen targets that meet the requirements.

[0084] Reference Figure 10 As shown, Figure 10 This is a detailed flowchart of the second embodiment of the microparticle printing method based on fluorescence field and bright field temporal imaging provided by the present invention. The microparticle printing method based on fluorescence field and bright field temporal imaging also includes, but is not limited to, steps S630 to S650, specifically: S630: After determining whether the multidimensional feature parameters meet the printing conditions, determine whether the number of target fluorescent particles printed meets the set threshold. S640: If the number of target fluorescent particles printed does not meet the set threshold, turn off the bright field illumination, restore the multicolor fluorescence field monitoring state, and wait for the next potential target to appear; S650: If the number of target fluorescent particles printed meets the set threshold, the printing operation ends.

[0085] Particle printing methods based on fluorescence field and bright field time-series imaging also include setting a threshold, which refers to the number of target fluorescent particles that need to be printed in the experiment or application. For example, in single-cell sorting experiments, it may be necessary to print a specific number of cells into microplates for subsequent analysis.

[0086] The system counts the number of printed target fluorescent particles in real time during the printing process, and updates the print count counter each time a target particle is successfully printed.

[0087] When the number of prints falls short of the set threshold, the system shuts off the bright-field illumination to conserve resources and reduce light damage. After shutting off the bright-field illumination, it immediately resumes multicolor fluorescence field monitoring, continuing fluorescence imaging of the microfluidic chip sorting area to search for the next potential target fluorescent microparticle to make up for the shortfall in prints.

[0088] The newly acquired fluorescence images are analyzed in real time by the image processing unit to identify potential targets and verify whether their characteristic parameters meet the printing conditions. This process is repeated cyclically until the number of prints reaches a set threshold. When the number of prints reaches the set threshold, the system confirms that the printing task is complete and ends the printing operation. A command is sent to stop the operation of the printing channel and shut down all related equipment (such as the pressure and motion control unit).

[0089] This mechanism not only improves the system's operational efficiency but also ensures precise control over the number of prints, meeting the specific needs of experiments or applications. This efficient screening and printing process has significant application value in fields such as biomedical research, cell analysis, and biochip manufacturing.

[0090] This invention also provides a microparticle printing system based on fluorescence field and bright field temporal imaging, used to execute the microparticle printing method based on fluorescence field and bright field temporal imaging described above. (Refer to...) Figure 11 As shown, the microparticle printing system based on fluorescence field and bright field temporal imaging includes a microfluidic chip, an optical imaging unit, an image processing unit, and a pressure and motion control unit. The microfluidic chip includes a sorting area and a printing channel. The sorting area holds the microparticle sample to be sorted, and the printing channel provides a flow path for the target fluorescent microparticles to be printed. The optical imaging unit includes a multi-wavelength LED light source and a camera. The multi-wavelength LED light source provides multi-color fluorescence field illumination, and the camera monitors the sorting area of ​​the microfluidic chip under continuous multi-color fluorescence field illumination and acquires fluorescence images of the sorting area in real time. When a target fluorescent microparticle is identified in the fluorescence image, bright field illumination is automatically triggered, and a bright field image is acquired. The image processing unit performs fusion analysis on the fluorescence image and the bright field image, extracts multi-dimensional feature parameters, and determines whether the multi-dimensional feature parameters meet the printing conditions according to preset multi-modal judgment rules. When the multi-dimensional feature parameters meet the printing conditions, a printing command is sent. The pressure and motion control unit responds to the printing command by driving the printing channel of the microfluidic chip to print the target fluorescent microparticles to the designated wells of the microplate.

[0091] Understandably, the microfluidic chip is the core component of the entire system, containing a sorting area and a printing channel. The sorting area holds the particle samples to be sorted, while the printing channel provides a flow path for target fluorescent particles that meet the printing conditions, allowing them to be accurately printed to the designated location. The optical imaging unit is responsible for providing illumination and image acquisition. A multi-wavelength LED light source provides multi-color fluorescence field illumination, capable of meeting the excitation requirements of different fluorescent particles. The camera monitors the sorting area and acquires fluorescence images in real time under continuous fluorescence field illumination. When a target fluorescent particle is identified, bright-field illumination is triggered and a bright-field image is acquired. This time-series imaging method can obtain both fluorescence and bright-field information of the particles.

[0092] The image processing unit fuses and analyzes the acquired fluorescence and bright-field images to extract multidimensional feature parameters, which may include the fluorescence intensity, color, shape, and size of the particles. Based on preset multimodal judgment rules, it determines whether the multidimensional feature parameters meet the printing conditions. If the conditions are met, a printing command is sent. Through sequential imaging of the fluorescence and bright-field fields, the target fluorescent particles can be accurately identified. Combined with the analysis of multidimensional feature parameters, this ensures that only particles meeting the conditions are printed, improving the accuracy and reliability of the printing process.

[0093] The pressure and motion control unit responds to the printing command and controls the printing channel of the microfluidic chip to accurately print the target fluorescent microparticles to the designated wells of the microplate.

[0094] This system combines advanced optical imaging and microfluidic technologies to provide an efficient and reliable method for the precise sorting and printing of microparticles, with broad application prospects. From particle identification and feature parameter extraction to the sending and execution of printing commands, the entire process can be automated, improving work efficiency and reducing human error.

[0095] It should be understood that the method steps in the embodiments of the present invention can be implemented or carried out by computer hardware, a combination of hardware and software, or by computer instructions stored in a non-transitory computer-readable storage medium. The method can use standard programming techniques. Each program can be implemented in a high-level procedural or object-oriented programming language to communicate with the computer system. However, if necessary, the program can be implemented in assembly or machine language. In any case, the language can be a compiled or interpreted language. Furthermore, for this purpose, the program can run on a programmed application-specific integrated circuit (ASIC).

[0096] Furthermore, the procedures described herein may be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by the context. The procedures described herein (or variations and / or combinations thereof) may be executed under the control of one or more computer systems configured with executable instructions, and may be implemented by hardware or a combination thereof as code (e.g., executable instructions, one or more computer programs, or one or more applications) that commonly executes on one or more processors. The computer program comprises a plurality of instructions executable by one or more processors.

[0097] Furthermore, the method can be implemented in any suitable type of computing platform, including but not limited to personal computers, minicomputers, mainframes, workstations, networked or distributed computing environments, standalone or integrated computer platforms, or in communication with charged particle tools or other imaging devices, etc. Aspects of the invention can be implemented as machine-readable code stored on a non-transitory storage medium or device, whether removable or integrated into a computing platform, such as a hard disk, optical read and / or write storage medium, RAM, ROM, etc., such that it is readable by a programmable computer, and when the storage medium or device is read by the computer, it can be used to configure and operate the computer to perform the processes described herein. Furthermore, the machine-readable code, or portions thereof, can be transmitted via wired or wireless networks. The invention described herein includes these and other different types of non-transitory computer-readable storage media when such media comprises instructions or programs that implement the steps described above in conjunction with a microprocessor or other data processor. When programmed according to the methods and techniques described in the invention, the invention may also include the computer itself.

[0098] A computer program can be applied to input data to perform the functions described herein, thereby transforming the input data to generate output data stored in non-volatile memory. The output information can also be applied to one or more output devices, such as a display. In a preferred embodiment of the invention, the transformed data represents physical and tangible objects, including specific visual depictions of physical and tangible objects generated on the display.

[0099] The above description is merely a preferred embodiment of the present invention. The present invention is not limited to the above-described embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention, as long as they achieve the technical effects of the present invention by the same means, should be included within the scope of protection of the present invention. Within the scope of protection of the present invention, the technical solutions and / or implementation methods can have various modifications and variations.

Claims

1. A microparticle printing method based on fluorescence field and bright field temporal imaging, characterized in that, include: Under continuous illumination of a multicolor fluorescence field, the sorting area of ​​the microfluidic chip is monitored, and fluorescence images of the sorting area of ​​the microfluidic chip are acquired in real time. When the target fluorescent particles are identified in the fluorescence image, bright-field illumination is automatically triggered, and a bright-field image is acquired. The fluorescence image and the bright field image are fused and analyzed, and multidimensional feature parameters are extracted. The multidimensional feature parameters are determined according to the preset multimodal judgment rules to see if they meet the printing conditions. When the multidimensional feature parameters meet the printing conditions, a printing command is sent. In response to the printing command, the pressure and motion control unit drives the printing channel of the microfluidic chip to print the target fluorescent microparticles onto the designated wells of the microplate.

2. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 1, characterized in that, The process of fusing and analyzing the fluorescence image and the bright-field image, and extracting multidimensional feature parameters, includes: The fluorescence image is analyzed and processed by the image processing unit to obtain the target feature parameters of the fluorescence image, including the fluorescence intensity and fluorescence color of the target fluorescent particles; The bright-field image is analyzed and processed by the image processing unit to obtain the target morphology parameters of the bright-field image, including the size of the target fluorescent particles and the roundness of the target fluorescent particles.

3. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 2, characterized in that, The step of determining whether the multidimensional feature parameters meet the printing conditions according to the preset multimodal judgment rules includes: When the fluorescence intensity of the target fluorescent particles is within a preset range and the fluorescence color of the target fluorescent particles is consistent with the set color, it is determined that the target feature parameters of the fluorescence image satisfy the preset multimodal judgment rule; When the size of the target fluorescent microparticle is within a preset range and the roundness of the target fluorescent microparticle is within a preset range, it is determined that the target morphology parameters of the bright field image satisfy the preset multimodal judgment rule; When the target feature parameters of the fluorescence image and the target morphology parameters of the bright field image simultaneously satisfy the preset multimodal judgment rule, it is determined whether the multidimensional feature parameters meet the printing conditions.

4. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 1, characterized in that, Also includes: The fluorescence image is uploaded to the image processing unit; The image processing unit performs real-time processing on the fluorescence image based on a recognition algorithm to identify potential targets; Determine whether the potential target meets the preset verification conditions; When the potential target meets the preset verification conditions, the potential target is identified as the target fluorescent microparticle.

5. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 1, characterized in that, The multicolor fluorescence field adopts a multicolor LED array and filter wheel scheme, and realizes intelligent switching of multicolor fluorescence through timing control and spectral matching; The multi-color LED array includes a three-color LED array composed of ultraviolet, blue, and green LEDs. The filter wheel is driven by a stepper motor and completes the filter switching within a preset time to uniformly collect fluorescence signals under multi-wavelength excitation.

6. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 1, characterized in that, Also includes: If the multidimensional feature parameters do not meet the printing conditions, immediately turn off the bright field illumination, restore the multicolor fluorescence field monitoring state, and wait for the next potential target to appear.

7. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 3, characterized in that, Also includes: After determining whether the multidimensional feature parameters meet the printing conditions, it is determined whether the number of target fluorescent particles printed meets the set threshold. If the number of printed target fluorescent particles does not meet the set threshold, the bright field illumination is turned off, and the system returns to the multicolor fluorescence field monitoring state, waiting for the next potential target to appear. If the number of target fluorescent particles printed meets the set threshold, the printing operation ends.

8. The microparticle printing method based on fluorescence field and bright field temporal imaging according to claim 1, characterized in that, Also includes: Once the printing operation is complete, turn off the bright field illumination and return to the multicolor fluorescence field monitoring state, waiting for the next potential target to appear.

9. A microparticle printing system based on fluorescence field and bright field temporal imaging, used to perform the microparticle printing method based on fluorescence field and bright field temporal imaging as described in any one of claims 1 to 8, characterized in that, include: A microfluidic chip includes a sorting area and a printing channel. The sorting area is used to hold a microparticle sample to be sorted, and the printing channel is used to provide a flow path for the target fluorescent microparticles to perform the printing operation. An optical imaging unit includes a multi-wavelength LED light source and a camera. The multi-wavelength LED light source is used to provide multi-color fluorescence field illumination. The camera is used to monitor the sorting area of ​​the microfluidic chip under continuous multi-color fluorescence field illumination and to acquire fluorescence images of the sorting area of ​​the microfluidic chip in real time. When a target fluorescent particle is identified in the fluorescence image, bright field illumination is automatically triggered and a bright field image is acquired. The image processing unit is used to perform fusion analysis on the fluorescence image and the bright field image, extract multi-dimensional feature parameters, determine whether the multi-dimensional feature parameters meet the printing conditions according to the preset multi-modal judgment rules, and send a printing command when the multi-dimensional feature parameters meet the printing conditions. A pressure and motion control unit is used to drive the printing channel of the microfluidic chip to print the target fluorescent microparticles to the designated wells of the microplate in response to the printing command.

10. A computer device comprising a memory and a processor, characterized in that, When the processor executes a computer program stored in the memory, it performs the method as described in any one of claims 1 to 8.

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