Living organism screening device and method based on fluorescence microscope

Abstract

The invention relates to the technical field of biological screening, and discloses a living organism screening device and method based on a fluorescence microscope. The injection pump is communicated with the sample introduction pool through a first conveying pipe; the buffer pool is communicated with the third electromagnetic valve; the stereoscope is communicated with the second electromagnetic valve through a second conveying pipe; the two-dimensional electric sorting table is communicated with the stereoscope through a third conveying pipe; the control module controls the flow speed of the first conveying pipe, the second conveying pipe and the third conveying pipe according to detection results of the first detector, the second detector and the third detector and controls opening and closing of the first electromagnetic valve, the second electromagnetic valve and the third electromagnetic valve. Sample images of the sight glass are collected, and the two-dimensional electric sorting table is controlled to classify and place the samples according to the sample images. The device provided by the invention realizes efficient and accurate screening of living biological samples through flow control design, control strategy and image analysis.

Description

Technical Field

[0001] The present invention relates to the field of biological screening technology, and in particular to a living organism screening device and method based on a fluorescence microscope. Background Art

[0002] The heterogeneity of biological samples often requires the collection of experimental results from a large number of samples for statistical analysis, as achieving rapid, large-scale imaging has become a key issue for microscopic imaging. Millimeter-scale biological samples, such as cell clusters, organoids, and micro-model organisms, are particularly vulnerable to high-throughput imaging due to their unique advantages in genetic development and drug screening research. For example, the zebrafish, a typical model animal, is widely used in developmental biology and drug screening due to its 87% genetic homology with humans, making it a top choice for high-throughput imaging.

[0003] The most direct approach to two-dimensional, high-throughput imaging of large, millimeter-sized samples, including zebrafish, is to embed the embryos in multi-well plates and image specific regions of the sample within each well using a scanning stage. This so-called high-content imaging approach has already been commercialized in several mature products. However, for samples with posture-oriented structures, such as nematodes and zebrafish, batch imaging of specific regions requires embryo alignment before the imaging experiment. While initial alignment of typical postures can be achieved using gel-pressing molds, meticulous manual manipulation is still required. To a certain extent, this imaging approach is still semi-automated, and sample recovery after imaging is difficult. The most prominent example of high-throughput technology in biological research tools is flow cytometry. While traditional flow cytometers can only detect fluorescence intensity, advances in imaging technology and their integration with fluidics have led to the emergence of high-throughput flow cytometry systems capable of two-dimensional imaging. These new flow cytometry systems enable rapid, large-scale imaging and high-throughput detection, providing more precise and accurate technical tools for biological research. The Yanik group at MIT expanded flow cytometry to image samples at the millimeter scale, reporting a high-throughput screening system that fully automates operations such as loading, positioning, and rotating zebrafish larvae, as well as image acquisition and data processing. Loading, imaging, and sorting can be completed in just 19 seconds. Subsequently, upgraded equipment was developed, but challenges arose in terms of sample size and cost, making it unsuitable for laboratory use.

[0004] Therefore, it is necessary to provide a living organism screening device and method based on a fluorescence microscope to solve the problem of low screening efficiency in the prior art. Summary of the Invention

[0005] In view of this, the present invention proposes a living organism screening device and method based on a fluorescence microscope, aiming to solve the problem of low screening efficiency in the prior art.

[0006] In one aspect, the present invention provides a living organism screening device based on a fluorescence microscope, comprising:

[0007] A sampling pool is provided for placing the sample to be screened, and the sampling pool is further connected to an air inlet pipe, on which an air pump and a first solenoid valve are provided. The air pump ventilates the sampling pool to prevent sample deposition in the sampling pool;

[0008] A syringe pump is connected to the sample injection pool through a first delivery tube, wherein the first delivery tube is sequentially provided with a second solenoid valve, a first detector and a third solenoid valve from the sample injection pool to the syringe pump, and the syringe pump is used to draw the sample from the sample injection pool;

[0009] a buffer tank, connected to the third solenoid valve, wherein a buffer solution is placed in the buffer tank;

[0010] a stereoscope, connected to the second solenoid valve via a second delivery pipe, wherein the second delivery pipe is further provided with a second detector;

[0011] a two-dimensional electric sorting platform, connected to the stereoscope via a third conveying pipe, wherein the third conveying pipe is provided with a third detector;

[0012] The control module is respectively connected to the sample injection pool, air pump, first solenoid valve, injection pump, first delivery pipe, second solenoid valve, first detector, third solenoid valve, buffer pool, stereoscope, second delivery pipe, second detector, two-dimensional electric sorting platform, third delivery pipe, and third detector. The control module controls the flow rate of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, and controls the opening and closing of the first solenoid valve, the second solenoid valve, and the third solenoid valve; the control module is also used to collect sample images of the stereoscope, and control the two-dimensional electric sorting platform to classify and place the samples according to the sample images.

[0013] Furthermore, when the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, it includes:

[0014] The syringe pump draws the sample into the flow channel, and the liquid flow rate in the flow channel is set to a first preset flow rate V1;

[0015] After the first detector detects the sample, the syringe pump is controlled to switch the flow rate in the flow channel to a second preset flow rate V2;

[0016] After the second detector detects the sample, the syringe pump is controlled to switch the flow rate in the switching flow channel to a third preset flow rate V3;

[0017] When the sample reaches the stereoscope, the syringe pump stops operating and the light source of the third detector is turned off, and the stereoscope collects the sample image;

[0018] Among them, the first preset flow rate V1> the second preset flow rate V2> the third preset flow rate V3 in the preset flow rates at different stages; the flow channel includes a first delivery pipe, a second delivery pipe and a third delivery pipe, and the flow rate of the flow channel is controlled by the injection pump.

[0019] Furthermore, when the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, it further includes:

[0020] The real-time flow rate in the flow channel is collected, and the real-time flow rate is compared with the preset flow rate of the stage, and it is determined whether the working state of the injection pump should be adjusted according to the comparison result.

[0021] Furthermore, the real-time flow rate in the acquisition flow channel is compared with the preset flow rate in the stage, and when determining whether to adjust the working state of the injection pump according to the comparison result, it includes:

[0022] Setting a preset flow rate threshold, if the absolute value of the difference between the real-time flow rate and the preset flow rate at this stage is less than the flow rate threshold, then determining not to adjust the working state of the injection pump;

[0023] If the absolute value of the difference between the real-time flow rate and the preset flow rate for this stage is greater than or equal to the flow rate threshold, it is determined that the working state of the injection pump is to be adjusted.

[0024] Furthermore, if the absolute value of the difference between the real-time flow rate and the preset flow rate in this stage is greater than or equal to the flow rate threshold, then the determination of adjusting the working state of the injection pump includes:

[0025] Calculate the difference between the real-time flow rate and the preset flow rate for the stage, and if the difference is greater than zero, record it as the first difference;

[0026] If the difference is less than zero, it is recorded as the second difference;

[0027] The working state of the injection pump is adjusted according to the magnitude of the first difference and the second difference.

[0028] Furthermore, the adjusting the working state of the injection pump according to the magnitude of the first difference and the second difference includes:

[0029] Setting a first difference limit and a second difference limit, wherein the first difference limit is smaller than the second difference limit;

[0030] If the first difference is less than or equal to the first difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient;

[0031] If the first difference is greater than the first difference limit and less than or equal to the second difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient;

[0032] If the first difference is greater than the second difference limit, adjusting the current flow rate of the injection pump by a third adjustment coefficient;

[0033] The value range of the adjustment coefficient is 1>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>0.5, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

[0034] Furthermore, when adjusting the working state of the injection pump according to the magnitude of the first difference and the second difference, the method further includes:

[0035] Setting a third difference limit and a fourth difference limit, wherein the third difference limit is smaller than the fourth difference limit;

[0036] If the second difference is less than or equal to the third difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient;

[0037] If the second difference is greater than the third difference limit and less than or equal to the fourth difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient;

[0038] If the second difference is greater than the fourth difference limit, adjusting the current flow rate of the syringe pump by a third adjustment coefficient;

[0039] The adjustment coefficient has a value range of 1.5>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>1, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

[0040] Furthermore, the control module is also used to collect sample images from a stereoscope, and control the two-dimensional electric sorting table to classify and place samples according to the sample images, including:

[0041] labeling fluorescent and non-fluorescent images according to sample images;

[0042] The samples corresponding to the fluorescent images and non-fluorescent images were classified and placed.

[0043] Furthermore, the sample concentration in the injection pool is 1 to 2 samples / ml;

[0044] The first detector, the second detector and the third detector are all optical fiber sensors.

[0045] Compared with the prior art, the present invention offers the following advantages: First, the device utilizes a fully automated fluidic system design. Through the coordinated operation of the sample injection reservoir, syringe pump, buffer reservoir, and stereoscope, it achieves automatic sample delivery and positioning, significantly improving screening efficiency. The air pump and first solenoid valve in the sample injection reservoir effectively prevent sample sedimentation, ensuring uniform sample suspension and providing reliable assurance for subsequent accurate detection. Second, the device utilizes three detectors (first, second, and third) combined with a multi-stage flow rate control strategy to precisely control the sample's movement within the delivery tube, ensuring smooth sample delivery to the detection position and avoiding the common problems of inaccurate positioning and sample damage associated with traditional manual operation. Specifically, by setting different preset flow rates, combined with real-time flow rate monitoring and dynamic adjustment, the sample delivery speed can be flexibly adjusted according to actual needs, ensuring both high-throughput screening efficiency and detection accuracy. Furthermore, the control module not only precisely controls the opening and closing of the solenoid valve and the flow rate of the syringe pump based on detector signals, but also enables automatic sample classification through image acquisition and analysis. This intelligent sorting based on fluorescence signals greatly reduces manual intervention, reduces human errors, and improves the reliability and repeatability of experimental results. The introduction of a two-dimensional electric sorting table further enhances the practicality of the device, enabling samples of different categories to be quickly and accurately sorted to designated locations. Finally, the modular design of the device gives it good scalability and adaptability, making it compatible with different types of fluorescence microscopes and biological samples. In general, the device achieves efficient and accurate screening of living biological samples through fluidic design, control strategies, and image analysis technology.

[0046] On the other hand, the present application also provides a living organism screening method based on fluorescence microscopy, comprising:

[0047] Place the sample to be screened in the injection pool and turn on the air pump to prevent sedimentation;

[0048] The syringe pump controls the flow rate of the flow channel to absorb the sample at a first preset flow rate V1. After the first detector detects the sample, the flow rate of the flow channel is reduced to a second preset flow rate V2. After the second detector detects the sample, the flow rate of the flow channel is reduced to a third preset flow rate V3.

[0049] Mark the fluorescent image and the non-fluorescent image according to the sample image, and classify and place the samples corresponding to the fluorescent image and the non-fluorescent image.

[0050] It is understandable that the living organism screening device and method based on fluorescence microscopy provided in this application have the same beneficial effects and will not be described in detail here. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Various other advantages and benefits will become apparent to those skilled in the art upon reading the detailed description of the preferred embodiment below. The accompanying drawings are for illustration purposes only and are not to be considered as limiting the present invention. The same reference symbols are used throughout the drawings to represent the same components. In the drawings:

[0052] Figure 1 A structural diagram of a living organism screening device based on a fluorescence microscope provided in an embodiment of the present invention;

[0053] Figure 2 A flow chart of a living organism screening method based on fluorescence microscopy provided in an embodiment of the present invention;

[0054] Figure 3 This is a timing diagram of synchronous control of the control module provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0055] Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present disclosure are shown in the accompanying drawings, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to enable a more thorough understanding of the present disclosure and to fully convey the scope of the present disclosure to those skilled in the art. It should be noted that, unless there is a conflict, the embodiments of the present disclosure and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in conjunction with the embodiments.

[0056] In some embodiments of the present application, see Figure 1 As shown, this embodiment provides a living organism screening device based on a fluorescence microscope, comprising:

[0057] A sampling pool is provided for placing the sample to be screened, and the sampling pool is further connected to an air inlet pipe, on which an air pump and a first solenoid valve are provided. The air pump ventilates the sampling pool to prevent sample deposition in the sampling pool;

[0058] A syringe pump is connected to the sample injection pool through a first delivery tube, wherein the first delivery tube is sequentially provided with a second solenoid valve, a first detector and a third solenoid valve from the sample injection pool to the syringe pump, and the syringe pump is used to draw the sample from the sample injection pool;

[0059] a buffer tank, connected to the third solenoid valve, wherein a buffer solution is placed in the buffer tank;

[0060] a stereoscope, connected to the second solenoid valve via a second delivery pipe, wherein the second delivery pipe is further provided with a second detector;

[0061] a two-dimensional electric sorting platform, connected to the stereoscope via a third conveying pipe, wherein the third conveying pipe is provided with a third detector;

[0062] The control module is respectively connected to the sample injection pool, air pump, first solenoid valve, injection pump, first delivery pipe, second solenoid valve, first detector, third solenoid valve, buffer pool, stereoscope, second delivery pipe, second detector, two-dimensional electric sorting platform, third delivery pipe, and third detector. The control module controls the flow rate of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, and controls the opening and closing of the first solenoid valve, the second solenoid valve, and the third solenoid valve; the control module is also used to collect sample images of the stereoscope, and control the two-dimensional electric sorting platform to classify and place the samples according to the sample images.

[0063] In some embodiments of the present application, when the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, the control module includes:

[0064] The syringe pump draws the sample into the flow channel, and the liquid flow rate in the flow channel is set to a first preset flow rate V1;

[0065] After the first detector detects the sample, the syringe pump is controlled to switch the flow rate in the flow channel to a second preset flow rate V2;

[0066] After the second detector detects the sample, the syringe pump is controlled to switch the flow rate in the switching flow channel to a third preset flow rate V3;

[0067] When the sample reaches the stereoscope, the syringe pump stops operating and the light source of the third detector is turned off, and the stereoscope collects the sample image;

[0068] Among them, the first preset flow rate V1> the second preset flow rate V2> the third preset flow rate V3 in the preset flow rates at different stages; the flow channel includes a first delivery pipe, a second delivery pipe and a third delivery pipe, and the flow rate of the flow channel is controlled by the injection pump.

[0069] In some embodiments of the present application, when the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, the control module further includes:

[0070] The real-time flow rate in the flow channel is collected, and the real-time flow rate is compared with the preset flow rate of the stage, and it is determined whether the working state of the injection pump should be adjusted according to the comparison result.

[0071] In some embodiments of the present application, the real-time flow rate in the flow channel is collected, and the real-time flow rate is compared with the preset flow rate in the stage, and whether to adjust the working state of the injection pump according to the comparison result includes:

[0072] Setting a preset flow rate threshold, if the absolute value of the difference between the real-time flow rate and the preset flow rate at this stage is less than the flow rate threshold, then determining not to adjust the working state of the injection pump;

[0073] If the absolute value of the difference between the real-time flow rate and the preset flow rate for this stage is greater than or equal to the flow rate threshold, it is determined that the working state of the injection pump is to be adjusted.

[0074] In some embodiments of the present application, if the absolute value of the difference between the real-time flow rate and the preset flow rate for the stage is greater than or equal to the flow rate threshold, then the determination of adjusting the working state of the syringe pump includes:

[0075] Calculate the difference between the real-time flow rate and the preset flow rate for the stage, and if the difference is greater than zero, record it as the first difference;

[0076] If the difference is less than zero, it is recorded as the second difference;

[0077] The working state of the injection pump is adjusted according to the magnitude of the first difference and the second difference.

[0078] In some embodiments of the present application, the adjusting the working state of the injection pump according to the magnitude of the first difference and the second difference includes:

[0079] Setting a first difference limit and a second difference limit, wherein the first difference limit is smaller than the second difference limit;

[0080] If the first difference is less than or equal to the first difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient;

[0081] If the first difference is greater than the first difference limit and less than or equal to the second difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient;

[0082] If the first difference is greater than the second difference limit, adjusting the current flow rate of the injection pump by a third adjustment coefficient;

[0083] The value range of the adjustment coefficient is 1>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>0.5, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

[0084] In some embodiments of the present application, when adjusting the working state of the injection pump according to the magnitude of the first difference and the second difference, the method further includes:

[0085] Setting a third difference limit and a fourth difference limit, wherein the third difference limit is smaller than the fourth difference limit;

[0086] If the second difference is less than or equal to the third difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient;

[0087] If the second difference is greater than the third difference limit and less than or equal to the fourth difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient;

[0088] If the second difference is greater than the fourth difference limit, adjusting the current flow rate of the syringe pump by a third adjustment coefficient;

[0089] The adjustment coefficient has a value range of 1.5>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>1, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

[0090] In some embodiments of the present application, the control module is further configured to collect a sample image from a stereoscope, and control the two-dimensional electric sorting table to classify and place the samples according to the sample image, including:

[0091] labeling fluorescent and non-fluorescent images according to sample images;

[0092] The samples corresponding to the fluorescent images and non-fluorescent images were classified and placed.

[0093] In some embodiments of the present application, the sample concentration in the injection pool is 1 to 2 samples / ml;

[0094] The first detector, the second detector and the third detector are all optical fiber sensors.

[0095] As can be understood, the device utilizes a fully automated fluidics system. Through the coordinated operation of the sample inlet reservoir, syringe pump, buffer reservoir, and stereoscope, it achieves automatic sample delivery and positioning, significantly improving screening efficiency. The air pump and first solenoid valve in the sample inlet reservoir effectively prevent sample sedimentation, ensuring uniform sample suspension and ensuring reliable subsequent detection. Secondly, the device utilizes three detectors (first, second, and third) coupled with a multi-stage flow rate control strategy to precisely control sample movement within the delivery tube, ensuring smooth sample delivery to the detection position and avoiding the positioning errors and sample damage common in traditional manual operations. Specifically, by setting different preset flow rates, combined with real-time flow rate monitoring and dynamic adjustment, the sample delivery speed can be flexibly adjusted according to actual needs, ensuring both high-throughput screening efficiency and detection accuracy. Furthermore, the control module not only precisely controls the opening and closing of the solenoid valve and the flow rate of the syringe pump based on detector signals, but also enables automatic sample classification through image acquisition and analysis. This intelligent sorting based on fluorescence signals greatly reduces manual intervention, reduces human errors, and improves the reliability and repeatability of experimental results. The introduction of a two-dimensional electric sorting table further enhances the practicality of the device, enabling samples of different categories to be quickly and accurately sorted to designated locations. Finally, the modular design of the device gives it good scalability and adaptability, making it compatible with different types of fluorescence microscopes and biological samples. In general, the device achieves efficient and accurate screening of living biological samples through fluidic design, control strategies, and image analysis technology.

[0096] Specifically, a zebrafish sample is placed into a sample injection pool at a concentration of approximately 1 sample / ml. An air pump is used to ventilate the sample to prevent sedimentation. A syringe pump draws the sample from the sample pool. When PD1 (the first detector) detects the sample, the direction of the second solenoid valve is changed, pushing the sample into the imaging pipeline (the second and third delivery tubes). The two detectors (the second and third detectors) are used for secondary deceleration. Once the sample is in place, images are captured. The samples are then sorted into the corresponding wells based on the results.

[0097] Specifically, taking zebrafish as an example, see Figure 3 , Figure 3 This is the timing diagram of the control module synchronization control.

[0098] (1) Initialize the control module, monitor the detector feedback signal in real time, and send instructions to the computer to control the liquid in the flow channel to be injected at a speed of V1, so that the sample passes through the flow channel at a uniform speed;

[0099] (2) When the first detector (PD1) detects the sample, the signal is fed back to the control module. After receiving the feedback signal, the control module switches the liquid in the flow channel to a speed of V2;

[0100] (3) When the second detector (PD2) detects the sample, the liquid in the flow channel is switched to V3 operation, and the control module performs a timer delay;

[0101] (4) The third detector (PD3) is used to monitor the accurate position of the sample. This position coincides with the center of the field of view. That is, after PD3 detects the sample, it is considered to have reached the final imaging area.

[0102] (5) When the sample reaches the focal plane of the objective lens (PD3 detects the sample), the syringe pump stops operating, the control module timer channel sends a trigger signal, and the camera starts image acquisition. Since the detection light emitted by the detector will affect the fluorescence imaging, the main control system will use the level signal to cooperate with the relay to realize the timely shutdown of the PD3 detector.

[0103] (6) The sample type is determined based on the collected fluorescence information. The stage is moved to the corresponding position based on the determined type, and the sample is ejected, thus completing a sample cycle.

[0104] On the other hand, see Figure 2 As shown, the present application also provides a living organism screening method based on a fluorescence microscope, which is applied to the above-mentioned living organism screening device based on a fluorescence microscope, comprising the following steps:

[0105] S100, placing the sample to be screened in the sampling pool and turning on the air pump to prevent sedimentation;

[0106] S200, the syringe pump controls the flow rate of the flow channel to aspirate the sample at a first preset flow rate V1, and after the first detector detects the sample, the flow rate of the flow channel is reduced to a second preset flow rate V2, and after the second detector detects the sample, the flow rate of the flow channel is reduced to a third preset flow rate V3;

[0107] S300 , marking fluorescent images and non-fluorescent images according to sample images, and placing samples corresponding to the fluorescent images and non-fluorescent images by classification.

[0108] It is understandable that the smooth transportation and precise positioning of samples are achieved through the multi-stage flow rate control strategy (V1>V2>V3), which not only ensures the efficiency of high-throughput screening, but also avoids damage or deviation of samples during high-speed flow. Secondly, the intelligent image recognition and classification system can accurately distinguish between fluorescent and non-fluorescent samples, greatly improving the accuracy and reliability of screening, and significantly improving work efficiency compared to traditional manual screening. The fully automated process adopted by this method reduces human intervention, which not only reduces operational errors, but also ensures the repeatability and consistency of experimental results. It is particularly worth mentioning that this method works perfectly with the aforementioned device to form a complete set of in vivo biological screening solutions, which achieves seamless connection from sample preparation to final sorting, and provides efficient and accurate technical support for developmental biology research, drug screening and other fields. In addition, the operating process of this method is simple and clear, easy to standardize and promote, and has great practical value and broad application prospects.

[0109] Those skilled in the art will appreciate that the embodiments of the present application may be provided as methods, systems, or computer program products. Therefore, the present application may take the form of an entirely hardware embodiment, an entirely software embodiment, or an embodiment combining software and hardware. Furthermore, the present application may take the form of a computer program product implemented on one or more computer-usable storage media (including but not limited to magnetic disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.

[0110] The present application is described with reference to the flowcharts and / or block diagrams of the methods, devices (systems) and computer program products according to the embodiments of the present application. It should be understood that each process and / or box in the flowchart and / or block diagram, as well as the combination of processes and / or boxes in the flowchart and / or block diagram, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, a special-purpose computer, an embedded processor or other programmable data processing device to produce a machine, so that the instructions executed by the processor of the computer or other programmable data processing device generate instructions for implementing the processes in the flowchart and / or block diagram. Figure 1 a process or multiple processes and / or boxes Figure 1 A device that provides the functions specified in a block or multiple blocks.

[0111] These computer program instructions may also be stored in a computer readable memory that can direct a computer or other programmable data processing device to work in a specific manner, so that the instructions stored in the computer readable memory produce an article of manufacture comprising an instruction device, which implements the process Figure 1 a process or multiple processes and / or boxes Figure 1 The function specified in one or more boxes.

[0112] These computer program instructions can also be loaded onto a computer or other programmable data processing device so that a series of operational steps are executed on the computer or other programmable device to produce a computer-implemented process, thereby providing the instructions executed on the computer or other programmable device for implementing the process. Figure 1 a process or multiple processes and / or boxes Figure 1 A step that specifies a function in one or more boxes.

[0113] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered by the scope of protection of the claims of the present invention.

Claims

1. A living organism screening device based on a fluorescence microscope, characterized in that: include: A sampling pool is provided for placing the sample to be screened, and the sampling pool is further connected to an air inlet pipe, on which an air pump and a first solenoid valve are provided. The air pump ventilates the sampling pool to prevent sample deposition in the sampling pool; A syringe pump is connected to the sample injection pool through a first delivery tube, wherein the first delivery tube is sequentially provided with a second solenoid valve, a first detector and a third solenoid valve from the sample injection pool to the syringe pump, and the syringe pump is used to draw the sample from the sample injection pool; a buffer tank, connected to the third solenoid valve, wherein a buffer solution is placed in the buffer tank; a stereoscope, connected to the second solenoid valve via a second delivery pipe, wherein the second delivery pipe is further provided with a second detector; a two-dimensional electric sorting platform, connected to the stereoscope via a third conveying pipe, wherein the third conveying pipe is provided with a third detector; The control module is respectively connected to the sample injection pool, air pump, first solenoid valve, injection pump, first delivery pipe, second solenoid valve, first detector, third solenoid valve, buffer pool, stereoscope, second delivery pipe, second detector, two-dimensional electric sorting platform, third delivery pipe, and third detector. The control module controls the flow rate of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, and controls the opening and closing of the first solenoid valve, the second solenoid valve, and the third solenoid valve; the control module is also used to collect sample images of the stereoscope, and control the two-dimensional electric sorting platform to classify and place the samples according to the sample images.

2. The living organism screening device based on fluorescence microscope according to claim 1, characterized in that: When the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, the control module includes: The syringe pump draws the sample into the flow channel, and the liquid flow rate in the flow channel is set to a first preset flow rate V1; After the first detector detects the sample, the syringe pump is controlled to switch the flow rate in the flow channel to a second preset flow rate V2; After the second detector detects the sample, the syringe pump is controlled to switch the flow rate in the switching flow channel to a third preset flow rate V3; When the sample reaches the stereoscope, the syringe pump stops operating and the light source of the third detector is turned off, and the stereoscope collects the sample image; Among them, the first preset flow rate V1> the second preset flow rate V2> the third preset flow rate V3 in the preset flow rates at different stages; the flow channel includes a first delivery pipe, a second delivery pipe and a third delivery pipe, and the flow rate of the flow channel is controlled by the injection pump.

3. The living organism screening device based on fluorescence microscope according to claim 2, characterized in that: When the control module controls the flow rates of the first delivery pipe, the second delivery pipe, and the third delivery pipe according to the detection results of the first detector, the second detector, and the third detector, the control module further includes: The real-time flow rate in the flow channel is collected, and the real-time flow rate is compared with the preset flow rate of the stage, and it is determined whether the working state of the injection pump should be adjusted according to the comparison result.

4. The living organism screening device based on fluorescence microscope according to claim 3, characterized in that: The real-time flow rate in the collecting flow channel is compared with the preset flow rate in the stage, and whether to adjust the working state of the injection pump according to the comparison result is determined, including: Setting a preset flow rate threshold, if the absolute value of the difference between the real-time flow rate and the preset flow rate at this stage is less than the flow rate threshold, then determining not to adjust the working state of the injection pump; If the absolute value of the difference between the real-time flow rate and the preset flow rate for this stage is greater than or equal to the flow rate threshold, it is determined that the working state of the injection pump is to be adjusted.

5. The living organism screening device based on fluorescence microscope according to claim 4, characterized in that: If the absolute value of the difference between the real-time flow rate and the preset flow rate for the stage is greater than or equal to the flow rate threshold, then the operating state of the injection pump is adjusted, including: Calculate the difference between the real-time flow rate and the preset flow rate for the stage, and if the difference is greater than zero, record it as the first difference; If the difference is less than zero, it is recorded as the second difference; The working state of the injection pump is adjusted according to the magnitude of the first difference and the second difference.

6. The living organism screening device based on fluorescence microscope according to claim 5, characterized in that: The adjusting of the working state of the injection pump according to the magnitude of the first difference and the second difference includes: Setting a first difference limit and a second difference limit, wherein the first difference limit is smaller than the second difference limit; If the first difference is less than or equal to the first difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient; If the first difference is greater than the first difference limit and less than or equal to the second difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient; If the first difference is greater than the second difference limit, adjusting the current flow rate of the injection pump by a third adjustment coefficient; The value range of the adjustment coefficient is 1>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>0.5, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

7. The living organism screening device based on fluorescence microscope according to claim 6, characterized in that: When the working state of the injection pump is adjusted according to the magnitude of the first difference and the second difference, the method further includes: Setting a third difference limit and a fourth difference limit, wherein the third difference limit is smaller than the fourth difference limit; If the second difference is less than or equal to the third difference limit, adjusting the current flow rate of the syringe pump by a first adjustment coefficient; If the second difference is greater than the third difference limit and less than or equal to the fourth difference limit, adjusting the current flow rate of the syringe pump by a second adjustment coefficient; If the second difference is greater than the fourth difference limit, adjusting the current flow rate of the syringe pump by a third adjustment coefficient; The adjustment coefficient has a value range of 1.5>first adjustment coefficient>second adjustment coefficient>third adjustment coefficient>1, and the adjusted injection pump flow rate is the product of the injection pump flow rate before adjustment and the adjustment coefficient.

8. The living organism screening device based on fluorescence microscope according to claim 1, characterized in that: The control module is also used to collect sample images from the stereoscope, and control the two-dimensional electric sorting table to classify and place samples according to the sample images, including: labeling fluorescent and non-fluorescent images according to sample images; The samples corresponding to the fluorescent images and non-fluorescent images were classified and placed.

9. The living organism screening device based on fluorescence microscope according to claim 1, characterized in that: The sample concentration in the injection pool is 1-2 samples / ml; The first detector, the second detector and the third detector are all optical fiber sensors.

10. A method for screening living organisms based on a fluorescence microscope, applied to the living organism screening device based on a fluorescence microscope as claimed in any one of claims 1 to 9, characterized in that: include: Place the sample to be screened in the injection pool and turn on the air pump to prevent sedimentation; The syringe pump controls the flow rate of the flow channel to absorb the sample at a first preset flow rate V1. After the first detector detects the sample, the flow rate of the flow channel is reduced to a second preset flow rate V2. After the second detector detects the sample, the flow rate of the flow channel is reduced to a third preset flow rate V3. Mark the fluorescent image and the non-fluorescent image according to the sample image, and classify and place the samples corresponding to the fluorescent image and the non-fluorescent image.

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