CN217425226U - Single cell capture system based on image analysis - Google Patents

Abstract

The utility model discloses a unicell 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 which is respectively connected with the sampling unit and the image analysis unit; 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 sucker and a sampling pipeline which are communicated; the sampling suction head sucks a target object from the liquid storage unit and transfers the target object through a sampling pipeline communicated with the sampling suction head; 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 for controlling the sampling sucker; the first displacement control module is connected with the image analysis unit; the first displacement control module controls the displacement of the sampling sucker according to the information fed back by the image analysis unit. The utility model provides a system is caught to unicell can high efficiency, high specificity realize the catch to unicell and analog.

Description

Single cell capture system based on image analysis

Technical Field

The utility model relates to a biomedical engineering technical field especially relates to a unicellular system of catching based on image analysis.

Background

The single cell sequencing technology is used for sequencing and quantifying information such as single cell genome, transcriptome and the like, so as to reveal cell population difference and cell evolution relation. The high-throughput Sequencing technology, that is, Next Generation Sequencing (NGS), can comprehensively and finely analyze genomes and transcriptomes of one species, can perform Sequencing on hundreds of thousands to millions of DNA molecules at one time in parallel, takes the reading length as a mark, and splices a plurality of short DNA fragments into complete sequence information; the method has remarkable advantages in processing large-scale samples and is a core technology in omics research at present.

In 1964, professor Robert Holley (Robert Holley) of cornell university, usa, invented the earliest sequencing technology. Thereafter, the development of molecular cloning, gel electrophoresis and autoradiography techniques has enabled the direct reading of DNA sequences to take place; wurui, a Chinese biologist of the university of Kannell, USA, used a primer extension method for DNA sequencing in 1971, and provides a technical basis for a Sanger sequencing method in the future; in 1975, the British biochemist Friderlike Mulger (Frederick Sanger) invented an epoch-making sequencing technology Sanger sequencing method in the field of life sciences on the basis of the Wurui sequencing method, and in 1977, the genome sequence of the phi X174 phage, which is the first complete genome complete sequence read by human, was successfully sequenced by using the technology. The first generation sequencing technology has the main characteristics that the sequencing reading length can reach 1000bp, the accuracy is as high as 99.999%, but the defects of high sequencing cost, low flux and the like seriously influence the real large-scale application of the technology.

With the proposal of Human Genome Project (HGP) in 1985, sequencing is rapidly developing towards larger sample size, more data size and more species, and the first generation sequencing technology cannot become an ideal sequencing method. Through continuous technical development and improvement, a high-throughput sequencing technology which takes 454 of Roche, Solexa and Hiseq technologies of illumina and Solid technologies of ABI as markers is produced, so that the sequencing cost is greatly reduced, the sequencing speed is greatly improved, the high accuracy is kept, and the sequence reading length is much shorter than that of the first generation sequencing technology.

With the rapid development of high-throughput sequencing technology, it is widely applied to various biological problems. De novo sequencing (de novo sequencing) is carried out on a species without a reference sequence on a genome level to obtain the reference sequence of the species, thereby laying a foundation for subsequent research and molecular breeding; carrying out whole genome re-sequencing (resequencing) on species with reference sequences, scanning and detecting mutation sites on the whole genome level, and finding out the molecular basis of individual difference; and the microbial diversity and the function identification can be carried out at the metagenome level. Performing whole transcriptome sequencing (white transcriptome sequencing) on a transcriptome level, so as to perform researches on alternative splicing, coding sequence single nucleotide polymorphism (cNPD) and the like; or small-molecule RNA sequencing (small RNA sequencing), and isolating RNA molecules of a specific size for sequencing, thereby discovering new microRNA molecules, and the like. At the apparent group level, combined with chromatin co-immunoprecipitation (ChIP) and methylated DNA co-immunoprecipitation (MeDIP) techniques, DNA regions binding to specific transcription factors and methylated sites on the genome, etc. are detected. At the proteome level, the method is used for identifying proteins, characterizing posttranslational modifications of the proteins, analyzing the relationship between the primary structure and the function of the proteins and the like. On the level of the metabolome, the metabolites in the organism are quantitatively analyzed, and the relative relationship between the metabolites and the physiological and pathological changes is searched. In addition, target sequencing technologies (Targeted sequencing) derived based on high throughput sequencing combined with microarray technology, such as whole exome capture sequencing, etc., are available. With the development of the technology, the sequencing technology is further advanced to the single cell level, so that the research on early embryonic development, cell heterogeneity, lineage path analysis and the like is facilitated. In addition, gene expression is both time-and space-specific, and new sequencing technologies can further resolve cell types and gene expression patterns in both the temporal and spatial dimensions.

The single cell sequencing technology is used for sequencing and quantifying information such as single cell genome, transcriptome and the like, so as to reveal cell population difference and cell evolution relation. In recent years, with the rapid development of high-throughput sequencing technology, the technology of omics sequencing on single cells is becoming mature, and the cost of single cell sequencing is decreasing. With the continuous and deep research, various single cell sequencing technologies have been developed, and the latest domestic and foreign research results have realized the comprehensive sequencing and exploration of whole genomes, transcriptomes, epigenetics, proteomics and the like at the single cell level. At present, single cell technology has been widely applied to basic and clinical research, and single cell-based omics sequencing has become one of the necessary means for research and exploration in many disciplines such as oncology, microbiology, neurobiology, reproductive medicine, immunology, and the like.

The inception of single cell sequencing technology dates back to 2006 at the earliest. By utilizing a multiple displacement amplification method, scientific research personnel amplify the Feike-level DNA to the microgram level, so that the threshold value of machine capture and sequencing is reached, and the door is opened for the detection of single cell level omics. In a short period of time thereafter, various sequencing techniques appeared as spring shoots after rain. In 2009, Tangfuyu et al established a single cell transcriptome sequencing method for the first time on the basis of the single cell genome amplification method, and laid the foundation for the subsequent single cell transcriptome research. In the next few years, single cell transcriptome sequencing technologies, such as STRT-seq, Smart-seq2, etc., were continuously improved in top-level journals. In addition to transcriptomes, epigenetic sequencing techniques have also been developed. The PBAT method and the scBS-seq method appear between 2012 and 2013, and the blank of single cell methylation sequencing is filled. In 2015, the first method for sequencing the open state of the chromosome of a single cell was published in Nature by utilizing the characteristic that DNase I can cut and digest the chromosome which is not coated by a nucleosome, which also indicates that the first research on epigenetics enters a brand new stage. Under the increasing demand of scientific research, the scheme of sequencing only on the simplex group cannot complete many scientific research explorations. In 2015, researchers at sanger institute took the lead to separate DNA and RNA by magnetic beads, and realized simultaneous detection of transcriptome and genome. The scM & T-seq technique developed by Wolf Reik team, Cambridge university, provides a reliable protocol for simultaneous detection of single cell transcriptomes and DNA methylation by combining Smart-seq2 and scBS-seq. The 17-year-old scNOME-seq can simultaneously determine the chromosome opening state and DNA methylation at the single cell level. CITE-seq and ECCITE-seq technologies developed by Satiji laboratories achieve simultaneous determination of a portion of cell surface proteins and transcriptomes of individual cells by a method of coupling a specific base tag to an antibody. In addition, techniques such as scTrio-seq, scCOOL-seq, developed by professor boreale et al, can simultaneously detect the genomic, DNA methylation, transcriptome/chromosome patency status of a single cell. In order to explore the spatial relationship of cells, transcript sequencing protocols such as Slide-seq, etc., can be developed in the spatial domain.

The currently mainstream single cell capture techniques include the following two:

1.10X Genomics based droplet capture technology: the individual cells were sorted by microfluidic technology, the beads were passed through the tube one by one under pressure, and the cells and enzymes were in another vertical tube, 1 cell was knocked onto 1 bead, which was then mixed into the oil phase. Single cells and beads are wrapped by oil drops to form a water-in-oil structure, and a microenvironment for adsorbing mRNA in 1 cell by 1 bead is created. Thereby achieving the microenvironment for capturing single cells and constructing subsequent libraries.

BD Rhapsody single cell capture system: BD Rhapbody utilizes microwells for single cell separation, belonging to Microwell-seq. The size of 20 ten thousand microwells is designed such that one microwell can exactly accommodate one bead to which a reverse transcription primer is attached. During capture, the single cell suspension is diluted to an appropriate concentration, the single cells are randomly captured by the micropores, the single cell capture rate is 80%, the multi-cell rate of 1K captured cells is about 0.2%, and the multi-cell rate of 10K captured cells is about 2.4%. Rhapbody suggests a detection flux of 100-.

The single cell sequencing technology is developed to the present, and dozens of single cell sequencing technologies with different application scenes have been developed in the global scope, but generally, the single cell sequencing technology is divided into three links: single cell capture, library construction, and high throughput next generation sequencing. At present, different library construction methods exist according to application scenes, high-throughput second-generation sequencing is also a very mature platform, and the most critical single-cell capture technology is the core for limiting the application of the high-throughput second-generation sequencing. The currently mainstream single cell capture technology platform includes related products based on microfluidics technology, and by using a microfluidics device, microbeads with barcodes, cells, and enzymes, primers and the like required by the reaction can be put into tiny droplets together, and the reaction is carried out in hundreds of thousands of droplets, so that thousands of single cell transcriptome libraries can be obtained in a short time, and a representative method includes the technology based on Drop-seq of the above-mentioned 10x Genomics company. The other single cell capturing technology platform is the Micorowell technology for capturing the high-flux cells through natural sedimentation, and single cells are settled into micropores capable of accommodating only one cell, so that a micro reaction system is formed for constructing the library. These mainstream single cell capture technologies still face more limitations, such as:

1) the single cell technology based on microfluidics has definite limitation on the cell size, the microfluidic chip of 10x Genomics generally requires that the cell diameter is less than 40um, and before a single cell sample is loaded on a machine, the cell must be filtered by a mesh screen to remove the cell with the diameter more than 40um, so that the problems of sample preference, sample information loss and the like can be caused. Meanwhile, too small cells or cell-like cells such as organelles and the like are difficult to capture, so that the application range of the technology is limited;

2) because the current mainstream method adopts a water-in-oil droplet form to form a micro reaction system for nucleic acid amplification, the method has certain preference on cell types, for example, the fat cells have low efficiency in the process of forming the water-in-oil droplets, and further influence the library building and amplification efficiency and the like.

3) At present, the mainstream microfluidic and Microwell technical systems directly crack cells in liquid drops or micropores, and then capture mRNA information by using a microbead coupled with a primer sequence barcode in the liquid drops or the micropores. Because the nucleoplasm separation of single cells cannot be realized, the single cell multi-omics library establishment and sequencing cannot be realized;

4) at present, the mainstream microfluidic and Microwell technology systems are limited in capture technology and sequencing depth, so that the number of genes which can be detected by finally obtained average single cells is small, partial sample information can be lost, and the deep data mining and analysis work at the later stage is not facilitated.

5) At present, the mainstream microfluidic and Microwell technology systems capture cells indiscriminately, and in order to improve the capture efficiency of living cells and produce high-quality sequencing data, the current capture technology systems require that the cell viability of the loaded single-cell suspension is detected firstly, the cell viability in the suspension needs to reach a certain standard, if the cell viability is more than 85%, indiscriminate capture is carried out later, and the captured apoptotic cells can be filtered only by a bioinformatics algorithm subsequently.

6) At present, the mainstream microfluidic and Microwell technology system adopts a strategy of mixing and integrally building a library of captured cells, and is limited by capture efficiency, so that the number of the loaded cells is large, at least more than hundreds of cells are needed, and generally tens of thousands of cells are needed, and the microfluidic and Microwell technology system is not suitable for samples with few cells, such as single-cell samples of single embryos.

SUMMERY OF THE UTILITY MODEL

To the technical problem mentioned above, the utility model provides a unicellular system of catching based on image analysis. The utility model provides a single cell capture system's capture principle is different from the single cell capture system of present mainstream, based on image analysis ization, carries out image analysis and with its two-dimensional positional information transmission with single cell or unicellular analogue distribution, activity or other characteristics, then absorbs one by one target unicellular or unicellular analogue in the target area, and the unicellular or unicellular analogue accessible sampling tube of absorption shifts and collects, and the follow-up storehouse operation of building that is used for multiple different usage.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

the utility model provides a single cell capture 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 respectively connected with the sampling unit and the image analysis unit;

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 sucker and a sampling pipeline which are communicated; the sampling sucker sucks a target object from the liquid storage unit and transfers the target object through a sampling pipeline communicated with the sampling sucker;

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 for controlling the sampling sucker; the first displacement control module is connected with the image analysis unit; and the first displacement control module controls the displacement of the sampling suction head according to the information fed back by the image analysis unit.

Preferably, the sampling unit comprises a plurality of groups of sampling suckers and sampling pipelines which are communicated in an isolated mode; the sampling suction head is detachably communicated with the sampling pipeline.

Preferably, the liquid storage unit is provided with a positioning mark, the positioning mark divides the liquid storage unit into a plurality of liquid storage areas, and the plurality of groups of sampling suction heads and sampling pipelines which are communicated in an isolated mode correspond to the plurality of liquid storage areas one to one.

Preferably, the light source and the image acquisition element are respectively arranged on the upper side and the lower side of the liquid storage unit.

Preferably, the device also comprises a sample introduction unit connected with the liquid storage unit, wherein the sample introduction unit comprises a sample introduction pipe and a sample spreader communicated with the sample introduction pipe; the sample applicator comprises a sample bin and a sample outlet; the sample outlet is arranged at the lower end of the sample bin; the section of the sample bin is convergent from the upper end to the sample outlet; and one end of the sample inlet pipe is provided with a sample inlet, and the other end of the sample inlet pipe is communicated with the sample bin.

Preferably, the device further comprises a collecting unit connected with the sampling pipeline and the control unit; the collecting unit comprises a porous 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 with the second displacement control module; the sample outlet needle tube is communicated with the sampling pipeline; the second displacement control module positions the sample outlet needle tubes to the target areas of the multi-hole culture plates by controlling the displacement of the sample outlet needle tubes on the multi-hole culture plates.

Preferably, the width of the sample compartment and the sample outlet corresponds to the width of the reservoir unit.

In the technical scheme of the present invention, the single cell analogue includes a cell nucleus, a cell organelle, and the like.

The technical scheme has the following advantages or beneficial effects:

1. the utility model provides a single cell capture system is unrestricted to the survival rate of last cell sample, the utility model discloses after carrying out image analysis location and discernment to the cell, absorb one by one with the suction head in order to catch, combine relevant technology such as trypan blue staining, can only absorb the live cell in the target area after image identification, therefore unrestricted and the requirement of the survival rate to last cell sample;

2. the single cell capturing system provided by the utility model has no preference for the size of the cell and no preference for the type of the cell; the single cell capturing system provided by the utility model adopts a single one-by-one single cell absorbing mode, so that no special requirements on the size of the cell are required, and the capturing from a large cell to a very small cell or even an organelle can be realized; meanwhile, the sucked single cells are collected by the collecting unit capable of directly separating samples without water-in-oil droplets, so that preference on cell types is avoided;

3. the utility model provides a single cell capture system can be to single cell or cell analogue image recognition one by one and catch, therefore theoretically do not have the lower limit to the cell quantity of computer sample, can catch as long as image recognition can, consequently can be used to the unicellular of few cell sample and catch;

4. in the prior art, a single cell is captured, a sample label technology of mixing a single cell sample and beads is used for capturing poly A tail of mRNA, thousands of cells are integrally built and sequenced at one time, the library construction flux is high, but the sequencing data is limited, the sequencing depth is low, and the number of genes measurable by a single cell is low; the single cell capturing system provided by the utility model can collect the captured single cells one by one and carry out the subsequent warehouse building treatment, thus being convenient for realizing the nuclear and cytoplasmic separation to carry out the single cell multi-omics warehouse building; meanwhile, because single cells are independently subjected to library building and sequencing, the sequencing depth is higher, the measurable basic factor of the single cells in the obtained data is 2-8 times higher than the average measured gene number of the single cells in the mainstream technology theoretically, and the high-depth single cell sequencing of multiple groups can be realized;

5. in the single-cell capturing system provided by the utility model, after the light source in the optical imaging unit is replaced by the laser emitter, the single-cell capturing system can be used for capturing specific fluorescent signal marked cells;

6. the utility model provides a unicellular system of catching through the suction head of changing different internal diameter specifications and the resolution ratio that improves the image analysis unit, can improve and catch the precision, not only can be used to the capture of unicellular, also can realize the capture of unicellular analogs such as littleer cell nucleus, subcellular organelle.

Drawings

The invention and its features, aspects and advantages will become more apparent from a reading of the following detailed description of non-limiting embodiments with reference to the attached drawings. Like reference symbols in the various drawings indicate like elements. The drawings are not intended to be drawn to scale, emphasis instead being placed upon illustrating the principles of the invention.

FIG. 1 is a schematic diagram of a single-cell trapping system according to example 1 of the present invention.

Fig. 2 is a schematic structural diagram of a sample injection unit of the single cell capture system in embodiment 1 of the present invention.

Fig. 3 is a schematic structural diagram of a sample injection unit of the single cell capture system in embodiment 1 of the present invention.

Detailed Description

In the following, the technical solutions in the embodiments of the present invention are clearly and completely described with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts belong to the protection scope of the present invention.

In the description of the present invention, it should be noted that, as the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. appear, the indicated orientation or positional relationship thereof is based on the orientation or positional relationship shown in the drawings, and is only for convenience of description and simplification of description, but does not indicate or imply that the indicated device or element must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.

In the description of the present invention, it is to be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" should be interpreted broadly, e.g., as being either fixedly connected, detachably connected, or integrally connected; they may be mechanically coupled, directly coupled, indirectly coupled through intervening media, or may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood as a specific case by those skilled in the art.

In the utility model, all the equipment and raw materials can be purchased from the market or commonly used in the industry if not specified. The methods in the following examples are conventional in the art unless otherwise specified.

Example 1:

this embodiment 1 provides a single-cell capturing system based on image analysis, as shown in fig. 1, including a liquid storage unit 1, an optical imaging unit 2, an image analysis unit 7, a sampling unit 3, and a control unit 4 respectively connected to the sampling unit 3 and the image analysis unit 7;

the optical imaging unit 2 is used for acquiring image information in the liquid storage unit 1 and comprises a light source 21 and an image acquisition element 22;

the sampling unit 3 comprises a sampling sucker 31 and a sampling pipeline 32 which are communicated; the sampling suction head 31 sucks the target object from the liquid storage unit 1, and transfers the target object through a sampling pipeline 32 communicated with the sampling suction head;

the image analysis unit 7 is connected with the image acquisition element 22 to receive and output image information;

the control unit 4 comprises a first displacement control module for controlling the sampling tip 31; the first displacement control module is connected with 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.

In the use process of the single cell capturing system provided in this embodiment, the liquid storage unit 1 is used to carry dissociated single cells or single cell analogs (such as nuclei, organelles, etc.) suspension (hereinafter referred to as cell suspension); the optical imaging unit 2 obtains image information of the cell suspension carried by the liquid storage unit 1 and then transmits the image information to the image analysis unit 7; the image analysis unit 7 analyzes the received image information and feeds the image information back to the control unit 4, controls the displacement of the sampling suction head 31 through the first displacement module, positions the sampling suction head to a target area of the liquid storage unit 1, sucks target single cells or single cell analogs from the liquid storage unit 1, and transfers the target single cells or single cell analogs through the sampling pipeline 32.

In order to further improve the operation convenience of the single cell capturing system, the sampling unit 3 comprises a plurality of groups of sampling suction heads 31 and sampling pipelines 32 which are communicated in an isolated way; the sampling tip 31 is detachably in communication with the sampling pipe 32. A plurality of groups of sampling suction heads 31 and sampling pipelines 32 which are communicated in an isolated way are arranged, so that a plurality of suction heads can synchronously work to suck a target single cell or a single cell analogue, the first displacement module independently controls the displacement of each sampling suction head, and the plurality of sampling suction heads are moved to different areas of the liquid storage unit 1 to synchronously suck, thereby further improving the sampling efficiency; and sampling suction head 31 detachably communicates with sampling pipe 32, can realize the convenient change of suction head, not only can change new suction head, also can change the suction head of different internal diameter specifications, has further improved the simple operation nature of unicellular capture system.

In order to further improve the practicability of the single cell capturing system, the liquid storage unit 1 is provided with positioning marks, the liquid storage unit 1 is divided into a plurality of liquid storage areas by the positioning marks, and the sampling suction heads 31 and the sampling pipelines 32 which are communicated in a multi-group isolated mode correspond to the plurality of liquid storage areas one to one. The liquid storage unit 1 is divided into a plurality of areas by positioning marks corresponding to different sampling suction heads, working areas of different sampling suction heads can be clearly displayed, and image information acquired by the optical imaging unit 2 is more accurate, so that the accuracy and the flux of the single cell capturing system can be improved. The positioning mark can adopt a positioning line, a positioning groove and the like.

In order to further improve the practicability of the single-cell capturing system, the light source 21 and the image capturing element 22 are respectively arranged at the upper side and the lower side of the liquid storage unit 1. In this embodiment, the upper and lower sides of the liquid storage unit 1 are not fixed with respect to the surface of the liquid storage unit carrying the cell suspension, the light source 21 and the image capturing element 22 are not fixed, when the light source 21 is located above, the image capturing element 22 is located below, and when the light source 21 is located below, the image capturing element 22 is located above. In the single cell capturing system provided by the utility model, the light emitted by the light source 21 irradiates the liquid storage unit 1 to generate optical image information; the image pickup element 22 observes and collects optical image information in a plurality of liquid storage regions of the liquid storage unit 1.

In order to further improve the operability of the single-cell capture system, the single-cell capture system further comprises a sample injection unit 5 connected to the stock solution unit 1, as shown in fig. 2, the sample injection unit 5 comprises a sample injection tube 51 and a sample applicator 52 communicated therewith; the sample applicator 52 includes a sample chamber and a sample outlet; the sample outlet is arranged at the lower end of the sample bin; the section of the sample bin is convergent from the upper end to the sample outlet; one end of the sample inlet pipe is provided with a sample inlet, and the other end of the sample inlet pipe is communicated with the sample bin. The utility model provides a unicellular capture system lays cell suspension in stock solution unit 1 through advance kind unit 5, in the use, cell suspension flows in the sample storehouse through the appearance pipe 51 of advancing, and sample laying ware 52 translates on stock solution unit 1, and the cell suspension flows stock solution unit 1 through the sample outlet that sample storehouse lower extreme set up under gravity or pressure, and the cross-section in sample storehouse sets up to the convergence form, can make cell suspension evenly drip, guarantees the homogeneity of cell suspension on the stock solution unit 1. The direction of movement of the sample applicator 52 on the reservoir unit 1 can be set arbitrarily, regardless of the direction of flow of the cell suspension into the sample inlet tube 51.

In order to further improve the practicability of the single cell capture system, the single cell capture system further comprises a collection unit 6 connected with the sampling pipeline 32 and the control unit 4; the collecting unit 6 comprises a porous culture plate 61 and a sample outlet needle tube 62; the control unit 4 further comprises a second displacement control module; the sample outlet needle tube 62 is connected with the second displacement control module; the sample outlet needle tube 62 is communicated with the sampling pipeline 32; the second displacement control module positions the sample-out needle 62 to a target area of the multi-well plate 61 by controlling the displacement of the sample-out needle 62 on the multi-well plate 61. The utility model provides a unicellular capture system, in the use, the unicellular or unicellular analog that sampling suction head 31 absorb flows into out appearance needle tubing 62 through sampling pipe 32, and the displacement of second displacement control module control appearance needle tubing 62 removes it to porous culture plate 61 target location to release unicellular or unicellular analog in the appearance needle tubing 62. The utility model discloses in, when sampling unit 3 set up the sampling suction head 31 and the sampling pipeline 32 of the isolated intercommunication of multiunit, a play appearance needle tubing of the isolated intercommunication of each sampling pipeline, the displacement of a plurality of play appearance needle tubing of second displacement control module independent control realizes a plurality of synchronous play appearance of play appearance needle tubing simultaneously, improves a kind efficiency.

To further improve the uniformity of the cell suspension on the reservoir unit, the width of the sample compartment and the sample outlet corresponds to the width of the reservoir unit 1. As shown in FIG. 3, in this embodiment, the movement direction of the sample applicator 5 on the reservoir unit 1 is shown by an arrow, the width of the reservoir unit 1 is the length perpendicular to the arrow on the plane on which the cell suspension is spread, and the widths of the sample chamber and the sample outlet, i.e., the length perpendicular to the arrow, are consistent with the length. The utility model provides a spread the appearance ware, spread the appearance in-process, along with the removal of spread the appearance ware, the cell suspension is followed the sample outlet and is dripped, the width that sets up the sample outlet is unanimous with the width of stock solution unit 1, can realize the cell suspension in bank and drip, the removal rate of spread the appearance ware keeps when reasonable within range, when the in-process of stock solution unit's one end translation to the other end is followed to the spread the appearance ware, the cell suspension evenly lays in the stock solution unit completely promptly, need not the repetitive movement operation, the homogeneity that the cell suspension distributes on the stock solution unit has been guaranteed.

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

Claims (7)

1. A single cell capturing system based on image analysis is characterized by comprising a liquid storage unit, an optical imaging unit, an image analysis unit, a sampling unit and a control unit which is respectively connected with the sampling unit and the image analysis unit;

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 sucker and a sampling pipeline which are communicated; the sampling suction head sucks a target object from the liquid storage unit and transfers the target object through a sampling pipeline communicated with the sampling suction head;

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 for controlling the sampling sucker; the first displacement control module is connected with the image analysis unit; and the first displacement control module controls the displacement of the sampling suction head according to the information fed back by the image analysis unit.

2. The single-cell capture system of claim 1, wherein the sampling unit comprises a plurality of sets of isolated communicating sampling tips and sampling conduits; the sampling suction head is detachably communicated with the sampling pipeline.

3. The single-cell capture system of claim 2, wherein the reservoir unit is provided with positioning marks, the positioning marks divide the reservoir unit into a plurality of reservoir regions, and the plurality of sets of isolated and communicated sampling tips and sampling conduits are in one-to-one correspondence with the plurality of reservoir regions.

4. The single-cell capture system of claim 1, wherein the light source and the image capture element are disposed on upper and lower sides of the reservoir unit, respectively.

5. The single-cell capture system of claim 1, further comprising a sample injection unit connected to the reservoir unit, the sample injection unit comprising a sample injection tube and a sample applicator in communication therewith; the sample applicator comprises a sample bin and a sample outlet; the sample outlet is arranged at the lower end of the sample bin; the cross section of the sample bin is convergent from the upper end to the sample outlet; one end of the sample inlet pipe is provided with a sample inlet, and the other end of the sample inlet pipe is communicated with the sample bin.

6. The single-cell capture system of claim 5, further comprising a collection unit coupled to the sampling conduit and the control unit; the collecting unit comprises a porous 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 with the second displacement control module; the sample outlet needle tube is communicated with the sampling pipeline; the second displacement control module positions the sample-discharging needle tubes to the target areas of the multi-hole culture plates by controlling the displacement of the sample-discharging needle tubes on the multi-hole culture plates.

7. The single-cell capture system of claim 6, wherein the width of the sample compartment and the sample outlet is the same as the width of the reservoir unit.

Images