A single cell capture system based on image analysis

Abstract

The application discloses a single cell capturing system based on image analysis, which comprises a liquid storage unit, an optical imaging unit, an image analysis unit, a sampling unit and a control unit connected with the sampling unit and the image analysis unit respectively; the optical imaging unit is used for acquiring image information in the liquid storage unit and comprises a light source and an image acquisition element; the sampling unit comprises a sampling suction head and a sampling pipeline in communication; the sampling suction head sucks target objects from the liquid storage unit and transfers the target objects through the sampling pipeline in communication; the image analysis unit is connected with the image acquisition element to receive and output image information; the control unit comprises a first displacement control module used for controlling the sampling suction head; the first displacement control module is connected with the image analysis unit; and the first displacement control module controls displacement of the sampling suction head according to information fed back by the image analysis unit. The single cell capturing system provided by the application can efficiently and specifically capture single cells and single cell analogs.

Description

Technical Field

[0001] This invention relates to the field of biomedical engineering technology, and in particular to a single-cell capture system based on image analysis. Background Technology

[0002] Single-cell sequencing technology refers to the sequencing and quantification of information such as the genome and transcriptome of a single cell, thereby revealing differences in cell populations and cell evolutionary relationships. High-throughput sequencing technology, also known as next-generation sequencing (NGS), can perform comprehensive and detailed analysis of the genome and transcriptome of a species. It can sequence hundreds of thousands to millions of DNA molecules in parallel at the same time, characterized by short read lengths, by reading multiple short DNA fragments and assembling them into complete sequence information. It has significant advantages in processing large-scale samples and is currently a core technology in omics research.

[0003] In 1964, Robert Holley, a professor at Cornell University in the United States, invented the earliest sequencing technology. Subsequently, the development of molecular cloning, gel electrophoresis, and autoradiography led to the development of direct reading of DNA sequences. In 1971, Wu Rui, a Chinese biologist at Cornell University, applied primer extension to DNA sequencing, providing the technological foundation for the later Sanger sequencing method. In 1975, British biochemist Frederick Sanger, building on Wu Rui's sequencing method, invented the epoch-making Sanger sequencing method in the life sciences, and in 1977, successfully sequenced the genome of bacteriophage ΦX174 using this technology—the first complete genome sequence of an organism deciphered by humans. The main characteristics of first-generation sequencing technology were read lengths up to 1000 bp and accuracy up to 99.999%, but its high cost and low throughput severely hampered its large-scale application.

[0004] With the launch of the Human Genome Project (HGP) in 1985, sequencing rapidly evolved towards larger sample sizes, more data volumes, and more species, rendering first-generation sequencing technologies inadequate. Through continuous technological development and improvement, high-throughput sequencing technologies, such as Roche's 454, Illumina's Solexa and HiSeq technologies, and ABI's Solid technology, emerged. These technologies significantly reduced sequencing costs while greatly increasing sequencing speed and maintaining high accuracy, although their read lengths were considerably shorter than those of first-generation sequencing technologies.

[0005] With the rapid development of high-throughput sequencing technology, it has been widely applied to various biological problems. At the genome level, de novo sequencing is performed on species for which no reference sequence exists, obtaining the reference sequence for subsequent research and molecular breeding. For species with reference sequences, whole-genome resequencing is performed to scan and detect mutation sites at the whole-genome level, revealing the molecular basis of individual differences. It can also be used at the metagenomic level for the identification of microbial diversity and function. At the transcriptome level, whole transcriptome resequencing is performed to conduct studies on alternative splicing, coding sequence single nucleotide polymorphisms (cSNPs), and small RNA sequencing, which involves sequencing RNA molecules of specific sizes to discover new microRNA molecules. At the epigenome level, it is combined with chromatin immunoprecipitation (ChIP) and methylated DNA immunoprecipitation (MeDIP) technologies to detect DNA regions that bind to specific transcription factors and methylation sites on the genome. At the proteomic level, sequencing is used to identify proteins, characterize post-translational modifications, and analyze the relationship between protein primary structure and function. At the metabolomic level, it enables quantitative analysis of metabolites in organisms and identifies the relative relationships between metabolites and physiological and pathological changes. Furthermore, there are targeted resequencing technologies derived from high-throughput sequencing combined with microarray technology, such as whole-exome capture sequencing. With technological advancements, sequencing technology has further progressed to the single-cell level, making it easier to explore early embryonic development, cellular heterogeneity, and analyze lineage pathways. Moreover, gene expression exhibits time and spatial specificity; new sequencing technologies can further elucidate cell types and gene expression patterns across time and space.

[0006] Single-cell sequencing technology refers to the quantification of genome, transcriptome, and other information in a single cell, thereby revealing differences in cell populations and cell evolutionary relationships. In recent years, with the rapid development of high-throughput sequencing technology, the technology for omics sequencing of single cells has become increasingly mature, and the cost of single-cell sequencing has been decreasing. As research continues to deepen, various single-cell sequencing technologies have been developed, and the latest research results both domestically and internationally have achieved comprehensive sequencing and exploration of the whole genome, transcriptome, epigenome, and proteomics at the single-cell level. Currently, single-cell technology is widely used in basic and clinical research, and single-cell-based omics sequencing has become an essential tool for scientific research in many disciplines such as oncology, microbiology, neurobiology, reproductive medicine, and immunology.

[0007] The rise of single-cell sequencing technology can be traced back to 2006. Using multiple substitution amplification, researchers amplified femtogram-level DNA to the microgram level, reaching the threshold for machine capture and sequencing, opening the door to single-cell omics detection. In the short term that followed, various sequencing technologies emerged rapidly. In 2009, Tang Fuchou et al., based on single-cell genome amplification methods, established the first single-cell transcriptome sequencing method, laying the foundation for subsequent single-cell transcriptome research. In the following years, single-cell transcriptome sequencing technology continued to improve, with methods such as STRT-seq, Smart-seq, and Smart-seq2 being published in top journals. Besides transcriptomics, epigenetic sequencing technologies have also made significant progress. Between 2012 and 2013, the emergence of PBAT and scBS-seq methods filled the gap in single-cell methylation sequencing. In 2015, utilizing the ability of DNase I to cleave and digest chromosomes not covered by nucleosomes, the first method for sequencing the open state of chromosomes in single cells was published in *Nature*, marking a new stage in epigenetics research. With increasing research demands, single-gene sequencing methods are no longer sufficient for many scientific explorations. In 2015, researchers at the Sanger Institute pioneered the separation of DNA and RNA using magnetic beads, enabling simultaneous detection of the transcriptome and genome. The scM&T-seq technology developed by Wolf Reik's team at the University of Cambridge, by combining Smart-seq2 and scBS-seq, provides a reliable method for simultaneously detecting single-cell transcriptomes and DNA methylation. The scNOME-seq technology, introduced in 2017, can simultaneously measure the open state of chromosomes and DNA methylation at the single-cell level. The CITE-seq and ECCITE-seq technologies developed by the Satiji laboratory, by conjugating specific base tags to antibodies, enable the simultaneous measurement of certain cell surface proteins and the transcriptome in single cells. In addition, technologies such as scTrio-seq and scCOOL-seq, developed by Professor Tang Fuchou's team at Peking University, can simultaneously detect single-cell genomes, DNA methylation, and transcriptome / chromosome open states. To explore spatial connections between cells, transcriptome sequencing solutions such as Slide-seq, capable of spatially distributed sequencing, have also emerged.

[0008] Currently, the mainstream single-cell capture technologies include the following two:

[0009] 1.10X Genomics' microfluidic droplet capture technology: Using microfluidics, single cells are sorted, and beads are passed one by one through channels under pressure. Cells and enzymes are in a separate vertical channel, where one cell is aspirated onto one bead, then mixed with an oil phase. The oil droplets encapsulate the single cell and bead, forming a water-in-oil structure, creating a microenvironment where one bead can adsorb the mRNA from one cell. This achieves the goal of capturing single cells and creating the microenvironment for subsequent library construction.

[0010] 2. BD Rhapsody Single-Cell Capture System: BD Rhapsody utilizes microwells for single-cell isolation, belonging to the Microwell-seq category. The 200,000 microwells are designed so that each well can precisely accommodate a single bead (magnetic bead) attached to a reverse transcription primer. During capture, the single-cell suspension is diluted to an appropriate concentration, and single cells are randomly captured by the microwells. The single-cell capture rate is 80%, the multi-cell ratio of capturing 1K cells is approximately 0.2%, and the multi-cell ratio of capturing 10K cells is approximately 2.4%. Rhapsody recommends a throughput of 100-10000.

[0011] Single-cell sequencing technology has evolved to the point where dozens of different applications have been developed globally. However, broadly speaking, single-cell sequencing technology consists of three stages: single-cell capture, library construction, and high-throughput next-generation sequencing (NGS). Currently, different library construction methods exist depending on the application, and high-throughput NGS is a very mature platform. The most critical single-cell capture technology is the core factor limiting its application. Currently, mainstream single-cell capture technology platforms include products based on microfluidics. Using microfluidic devices, barcode-labeled microbeads, cells, and necessary enzymes and primers are loaded into tiny droplets, and the reaction occurs within hundreds or thousands of droplets, resulting in thousands of single-cell transcriptome libraries in a short time. Representative methods include the Drop-seq technology from 10x Genomics mentioned above. Another single-cell capture technology platform is the Micorowell technology, which captures high-throughput cells through natural sedimentation. This technology allows single cells to settle into microwells that can only hold one cell, forming a micro-reaction system for library construction. Currently, these mainstream single-cell capture technologies still face many limitations, such as:

[0012] 1) Microfluidic-based single-cell technology has strict limitations on cell size. 10x Genomics' microfluidic chips generally require cells with a diameter of less than 40 μm. Single-cell samples must be filtered using a mesh screen before being processed to remove cells larger than 40 μm, which can lead to sample bias and loss of sample information. Furthermore, it is difficult to capture excessively small cells or cell-like structures, such as organelles, thus limiting the technology's application scope.

[0013] 2) Since the current mainstream method uses water-in-oil droplets to form a micro-reaction system for nucleic acid amplification, this method has a certain preference for cell types. For example, adipocytes are less efficient in forming water-in-oil droplets, which affects library construction and amplification efficiency.

[0014] 3) Currently, mainstream microfluidic and Microwell technologies involve directly lysing cells in droplets or microwells, followed by capturing mRNA information using a microbead coupled with a primer sequence barcode within the droplet or microwell. Because nucleocytoplasmic separation of single cells cannot be achieved, single-cell multi-omics library construction and sequencing are not feasible.

[0015] 4) Due to limitations in capture technology and sequencing depth, the mainstream microfluidics and Microwell technologies currently available have a limited number of genes that can be detected per cell on average. This may result in the loss of some sample information and hinder in-depth data mining and analysis in the later stages.

[0016] 5) Currently, mainstream microfluidics and Microwell technologies capture cells indiscriminately. In order to improve the capture efficiency of live cells and produce high-quality sequencing data, current capture technology systems require the single-cell suspension loaded with samples to first perform cell viability testing. The cell viability in the suspension must reach a certain standard, such as a cell viability of over 85%, before indiscriminate capture. For captured apoptotic cells, subsequent filtering can only be performed through bioinformatics algorithms.

[0017] 6) The current mainstream microfluidics and Microwell technology systems adopt the strategy of capturing cells and mixing them together for library construction. Due to the limitations of capture efficiency, they require a large number of cells to be loaded, at least several hundred cells, and generally tens of thousands of cells. Therefore, they are not suitable for samples with few cells, such as single-cell samples from single embryos. Summary of the Invention

[0018] To address the aforementioned technical problems, this invention provides a single-cell capture system based on image analysis. The capture principle of the single-cell capture system provided by this invention differs from current mainstream single-cell capture systems. Based on image analysis, it performs image analysis on the distribution, activity, or other characteristics of single cells or single-cell analogs and transmits their two-dimensional location information. Then, it sequentially captures target single cells or single-cell analogs within the target area. The captured single cells or single-cell analogs can be transferred and collected through sampling tubes for subsequent library construction operations for various purposes.

[0019] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0020] On one hand, the present invention provides a single-cell capture system based on image analysis, including a liquid storage unit, an optical imaging unit, an image analysis unit, a sampling unit, and a control unit connected to the sampling unit and the image analysis unit respectively;

[0021] The optical imaging unit is used to acquire image information within the liquid storage unit, including a light source and an image acquisition element;

[0022] The sampling unit includes a connected sampling tip and a sampling tube; the sampling tip draws the target substance from the liquid storage unit and transfers the target substance through the connected sampling tube;

[0023] The image analysis unit is connected to the image acquisition element to receive and output image information;

[0024] The control unit includes a first displacement control module for controlling the sampling tip; the first displacement control module is connected to the image analysis unit; the first displacement control module controls the displacement of the sampling tip according to the information fed back by the image analysis unit.

[0025] Preferably, the sampling unit includes multiple sets of isolated and connected sampling tips and sampling pipes; the sampling tips are detachably connected to the sampling pipes.

[0026] Preferably, the liquid storage unit is provided with positioning marks, which divide the liquid storage unit into multiple liquid storage areas, and the multiple sets of isolated and connected sampling tips and sampling pipes correspond one-to-one with the multiple liquid storage areas.

[0027] Preferably, the light source and the image acquisition element are respectively disposed on the upper and lower sides of the liquid storage unit.

[0028] Preferably, the system further includes a sample injection unit connected to the liquid storage unit. The sample injection unit includes a sample injection tube and a sample spreader connected thereto. The sample spreader includes a sample chamber and a sample outlet. The sample outlet is located at the lower end of the sample chamber. The cross-section of the sample chamber converges from the upper end to the sample outlet. One end of the sample injection tube has a sample inlet, and the other end is connected to the sample chamber.

[0029] Preferably, the system further includes a collection unit connected to the sampling conduit and the control unit; the collection unit includes a multi-well culture plate and a sample dispensing needle; the control unit further includes a second displacement control module; the sample dispensing needle is connected to the second displacement control module; the sample dispensing needle is in communication with the sampling conduit; the second displacement control module positions the sample dispensing needle to the target area of ​​the multi-well culture plate by controlling the displacement of the sample dispensing needle on the multi-well culture plate.

[0030] Preferably, the width of the sample chamber and the sample outlet is the same as the width of the liquid storage unit.

[0031] In another aspect, the present invention provides the use of the above-described single-cell capture system in capturing single cells and single-cell analogues.

[0032] In the technical solution of the present invention, the single-cell analogue includes the cell nucleus, organelles, etc.

[0033] The above technical solution has the following advantages or beneficial effects:

[0034] 1. The single-cell capture system provided by the present invention has no limit on the viability of single-cell samples. After image analysis, localization and identification of cells, the present invention uses a pipette tip to capture them one by one. Combined with trypan blue staining and other related technologies, after image identification, only live cells in the target area can be captured. Therefore, there are no restrictions or requirements on the viability of cell samples.

[0035] 2. The single-cell capture system provided by this invention has no preference for cell size or cell type; the single-cell capture system provided by this invention adopts a method of aspirating single cells one by one, so there are no special requirements for cell size, and it can capture large cells to extremely small cells and even organelles; at the same time, since the aspirated single cells are collected through a collection unit that can be directly sampled, without the need to pass through water-in-oil droplets, it has no preference for cell type.

[0036] 3. The single-cell capture system provided by the present invention can identify and capture individual cells or cell analogs one by one. Therefore, theoretically there is no lower limit to the number of cells in the sample. As long as the image is recognizable, it can be captured. Therefore, it can be used for single-cell capture of a very small number of cell samples.

[0037] 4. In existing technologies, after capturing single cells, sample tagging technology that mixes single-cell samples with beads is used to capture the poly A tail of mRNA. This allows for the simultaneous library construction and sequencing of thousands of cells, resulting in high throughput for library construction. However, due to limitations in sequencing data volume, the sequencing depth is relatively low, and the number of genes that can be sequenced per cell is relatively low. In contrast, the single-cell capture system provided by this invention allows for the individual collection of captured single cells for subsequent library construction, facilitating nucleocytoplasmic separation for single-cell multi-omics library construction. Furthermore, because individual cells are used for library construction and sequencing, the sequencing depth is higher. Theoretically, the number of genes that can be sequenced per cell in the obtained data is 2-8 times higher than the average number of genes sequenced per cell in mainstream technologies, enabling high-depth single-cell sequencing for multi-omics.

[0038] 5. In the single-cell capture system provided by the present invention, after the light source in the optical imaging unit is replaced with a laser emitter, it can be used to capture cells labeled with specific fluorescent signals;

[0039] 6. The single-cell capture system provided by the present invention can improve capture accuracy by changing the suction tip with different inner diameter specifications and improving the resolution of the image analysis unit. It can be used not only for capturing single cells, but also for capturing smaller single-cell analogues such as cell nuclei and subcellular organelles. Attached Figure Description

[0040] The invention, its features, shape, and advantages will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings. Like reference numerals denote like parts throughout the drawings. The drawings are not intentionally drawn to scale; the focus is on illustrating the spirit of the invention.

[0041] Figure 1 This is a schematic diagram of the single-cell capture system in Embodiment 1 of the present invention.

[0042] Figure 2 This is a schematic diagram of the sample introduction unit of the single-cell capture system in Embodiment 1 of the present invention.

[0043] Figure 3 This is a schematic diagram of the sample introduction unit of the single-cell capture system in Embodiment 1 of the present invention. Detailed Implementation

[0044] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Therefore, the detailed description of the embodiments of the present 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 present invention without inventive effort are within the protection scope of the present invention.

[0045] In the description of this invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. 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. Therefore, they should not be construed as limitations on this invention.

[0046] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; or they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0047] In this invention, unless otherwise specified, all equipment and raw materials are commercially available or commonly used in the industry. The methods described in the following embodiments are conventional methods in the art, unless otherwise specified.

[0048] Example 1:

[0049] This embodiment 1 provides a single-cell capture system based on image analysis, such as... Figure 1 As shown, it includes a liquid storage unit 1, an optical imaging unit 2, an image analysis unit 7, a sampling unit 3, and a control unit 4 that is connected to the sampling unit 3 and the image analysis unit 7 respectively;

[0050] The optical imaging unit 2 is used to acquire image information within the liquid storage unit 1, including a light source 21 and an image acquisition element 22;

[0051] The sampling unit 3 includes a sampling pipette 31 and a sampling tube 32 connected together; the sampling pipette 31 draws the target object from the liquid storage unit 1 and transfers the target object through the sampling tube 32 connected to it;

[0052] The image analysis unit 7 is connected to the image acquisition element 22 to receive and output image information;

[0053] The control unit 4 includes a first displacement control module for controlling the sampling tip 31; the first displacement control module is connected to the image analysis unit 7; the first displacement control module controls the displacement of the sampling tip 31 on the liquid storage unit 1 according to the information fed back by the image analysis unit 7.

[0054] In the single-cell capture system provided in this embodiment, during use, the liquid storage unit 1 is used to hold a suspension of dissociated single cells or single-cell analogs (such as cell nuclei, organelles, etc.) (hereinafter referred to as cell suspension); the optical imaging unit 2 acquires the image information of the cell suspension held in the liquid storage unit 1 and transmits it to the image analysis unit 7; the image analysis unit 7 analyzes the received image information and feeds it back to the control unit 4, and controls the displacement of the sampling pipette 31 through the first displacement module to position it to the target area of ​​the liquid storage unit 1, and draws the target single cell or single-cell analog from the liquid storage unit 1 and transfers it through the sampling pipe 32.

[0055] To further improve the ease of operation of the single-cell capture system, the sampling unit 3 includes multiple sets of isolated and interconnected sampling tips 31 and sampling tubing 32; the sampling tips 31 are detachably connected to the sampling tubing 32. By setting multiple sets of isolated and interconnected sampling tips 31 and sampling tubing 32, multiple tips can work simultaneously to aspirate target single cells or single-cell analogues. The first displacement module independently controls the displacement of each sampling tip, moving multiple sampling tips to different areas of the storage unit 1 for simultaneous aspiration, further improving sampling efficiency. Furthermore, the detachable connection of the sampling tips 31 to the sampling tubing 32 allows for convenient tip replacement, enabling not only the replacement of new tips but also the replacement of tips with different inner diameter specifications, further improving the ease of operation of the single-cell capture system.

[0056] To further improve the practicality of the single-cell capture system, positioning marks are provided on the liquid storage unit 1. These markings divide the liquid storage unit 1 into multiple liquid storage areas, with multiple sets of isolated and interconnected sampling tips 31 and sampling pipes 32 corresponding one-to-one with each of these areas. Dividing the liquid storage unit 1 into multiple areas corresponding to different sampling tips using positioning marks clearly displays the working area of ​​each tip, resulting in more accurate image information acquired by the optical imaging unit 2. This improves the accuracy and throughput of the single-cell capture system. The positioning marks can take the form of positioning lines or positioning grooves.

[0057] To further improve the practicality of the single-cell capture system, the light source 21 and the image acquisition element 22 are respectively disposed on the upper and lower sides of the liquid storage unit 1. In this embodiment, the upper and lower sides of the liquid storage unit 1 do not have a fixed orientation relative to the surface of the liquid storage unit that holds the cell suspension; when the light source 21 is above, the image acquisition element 22 is below, and vice versa. In the single-cell capture system provided by this invention, the light emitted by the light source 21 illuminates the liquid storage unit 1, generating optical image information; the image acquisition element 22 observes and collects optical image information from multiple liquid storage areas of the liquid storage unit 1.

[0058] To further improve the operability of the single-cell capture system, the single-cell capture system also includes a sample introduction unit 5 connected to the reservoir unit 1, such as... Figure 2 As shown, the sample injection unit 5 includes an injection tube 51 and a spreader 52 connected to it; the spreader 52 includes a sample chamber and a sample outlet; the sample outlet is located at the lower end of the sample chamber; the cross-section of the sample chamber converges from the upper end to the sample outlet; one end of the injection tube has an injection port, and the other end is connected to the sample chamber. The single-cell capture system provided by this invention spreads cell suspension onto the storage unit 1 through the sample injection unit 5. During use, the cell suspension flows into the sample chamber through the injection tube 51, and the spreader 52 moves horizontally on the storage unit 1. Under the action of gravity or pressure, the cell suspension flows onto the storage unit 1 through the sample outlet located at the lower end of the sample chamber. The cross-section of the sample chamber is designed to converge, which allows the cell suspension to drip evenly, ensuring the uniformity of the cell suspension on the storage unit 1. The direction of movement of the spreader 52 on the storage unit 1 can be arbitrarily set and is independent of the direction in which the cell suspension flows into the injection tube 51.

[0059] To further improve the practicality of the single-cell capture system, the system also includes a collection unit 6 connected to the sampling pipette 32 and the control unit 4. The collection unit 6 includes a multi-well culture plate 61 and a sample dispensing needle 62. The control unit 4 further includes a second displacement control module. The sample dispensing needle 62 is connected to the second displacement control module and communicates with the sampling pipette 32. The second displacement control module positions the sample dispensing needle 62 to the target area of ​​the multi-well culture plate 61 by controlling its displacement on the plate. In the single-cell capture system provided by this invention, during use, single cells or single-cell analogs drawn by the sampling pipette 31 flow into the sample dispensing needle 62 through the sampling pipette 32. The second displacement control module controls the displacement of the sample dispensing needle 62, moving it to the target position on the multi-well culture plate 61 and releasing the single cells or single-cell analogs from the needle 62. In this invention, when the sampling unit 3 is equipped with multiple isolated and connected sampling nozzles 31 and sampling channels 32, each sampling channel is isolated and connected to a sampling needle tube. The second displacement control module independently controls the displacement of multiple sampling needle tubes, and simultaneously realizes the synchronous sampling of multiple sampling needle tubes, thereby improving the sampling efficiency.

[0060] To further improve the uniformity of the cell suspension on the reservoir unit, the width of the sample chamber and sample outlet is the same as the width of reservoir unit 1. For example... Figure 3 As shown in the diagram, in this embodiment, the movement direction of the spreader 5 on the storage unit 1 is indicated by the arrow. The width of the storage unit 1 is its length perpendicular to the arrow direction on the plane where the cell suspension is spread. The widths of the sample chamber and the sample outlet are the same as this length, which is perpendicular to the arrow. In the spreader provided by this invention, during the spreader process, the cell suspension drips from the sample outlet as the spreader moves. By setting the width of the sample outlet to be consistent with the width of the storage unit 1, the cell suspension can be dripped in rows. When the moving speed of the spreader is kept within a reasonable range, the cell suspension is uniformly and completely spread on the storage unit as the spreader moves from one end to the other, eliminating the need for repeated movement operations and ensuring the uniformity of the cell suspension distribution on the storage unit.

[0061] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An image analysis-based single cell capture system, characterized by, The system comprises a liquid storage unit, an optical imaging unit, an image analysis unit, a sampling unit, and a control unit connected to the sampling unit and the image analysis unit respectively; The optical imaging unit is used to acquire image information in the liquid storage unit, and comprises a light source and an image acquisition element; The sampling unit comprises a sampling pipette and a sampling pipeline in communication; the sampling pipette sucks target objects from the liquid storage unit and transfers the target objects through the sampling pipeline in communication therewith; The image analysis unit is connected to the image acquisition element to receive and output image information; The control unit comprises a first displacement control module for controlling the sampling pipette; the first displacement control module is connected to the image analysis unit; the first displacement control module controls displacement of the sampling pipette according to information fed back by the image analysis unit; The system further comprises a sample feeding unit connected to the liquid storage unit; the sample feeding unit comprises a sample feeding pipeline and a sample spreader in communication therewith; the sample spreader comprises a sample chamber and a sample outlet; the sample outlet is arranged at a lower end of the sample chamber; a cross section of the sample chamber is convergent from an upper end to the sample outlet; one end of the sample feeding pipeline is provided with a sample inlet, and the other end is in communication with the sample chamber.

2. The single cell capture system of claim 1, wherein, The sampling unit comprises a plurality of groups of sampling pipettes and sampling pipelines in isolated communication; the sampling pipettes are detachably in communication with the sampling pipelines.

3. The single cell capture system of claim 2, wherein, The liquid storage unit is provided with positioning marks; the positioning marks divide the liquid storage unit into a plurality of liquid storage areas; the plurality of groups of sampling pipettes and sampling pipelines correspond to the plurality of liquid storage areas one by one.

4. The single cell capture system of claim 1, wherein, The light source and the image acquisition element are arranged on upper and lower sides of the liquid storage unit respectively.

5. The single cell capture system of claim 1, wherein, The system further comprises a collection unit connected to the sampling pipeline and the control unit; the collection unit comprises a multi-well culture plate and a sample outlet needle tube; the control unit further comprises a second displacement control module; the sample outlet needle tube is connected to the second displacement control module; the sample outlet needle tube is in communication with the sampling pipeline; the second displacement control module positions the sample outlet needle tube to a target area of the multi-well culture plate by controlling displacement of the sample outlet needle tube on the multi-well culture plate.

6. The single cell capture system of claim 1, wherein, The width of the sample chamber and the sample outlet is consistent with the width of the liquid storage unit.

7. Use of the single cell capturing system according to any one of claims 1-6 in capturing single cells and single cell analogs.

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