High-throughput living cell imaging analysis system

Abstract

The utility model discloses a high-throughput living cell imaging analysis system, which comprises a bottom plate and a sample table above the bottom plate, a first platform is slidably arranged at the top of the bottom plate along the X direction, the first platform is driven to slide by a first lead screw motor, a second platform is slidably arranged on the first platform along the Y direction, and the second platform is driven to slide by a second lead screw motor. A vertical sliding block is arranged at the top of the second platform in a sliding mode in the Z direction, the vertical sliding block is driven by a precise lead screw motor to slide, and a fluorescence imaging mechanism is arranged on the vertical sliding block. The whole set of instrument can be placed in a CO2 culture box, so that long-time living cell imaging of cells can be realized in a proper culture state. Different experiment conditions can be independently set for each cell culture container, and various experiments can be simultaneously realized in the same environment.

Description

Technical Field

[0001] This invention belongs to the field of cell technology, specifically a high-throughput live cell imaging analysis system. Background Technology

[0002] Cells, as the fundamental research subject in life sciences, are widely used in life sciences, drug development, and disease research. Researchers need to observe and study the growth status, internal mechanisms, drug responses, and gene transfection efficiency of cultured cells over long periods and in large quantities.

[0003] In life sciences and drug development, long-term, high-throughput observation of cell growth and drug responses is required. Traditional methods use microscopes to manually take periodic photographs for analysis, which is inefficient and cannot achieve high-throughput cell analysis. While live-cell imaging systems capable of continuous imaging exist, they typically only photograph one sample container at a time, failing to enable ultra-high-throughput sample analysis of multiple containers. Furthermore, existing such devices often only have one or two objectives, limiting the magnification for cell analysis. Utility Model Content

[0004] The purpose of this invention is to provide a high-throughput live cell imaging analysis system to solve the problems mentioned in the background art.

[0005] The technical solution adopted in this utility model is as follows:

[0006] A high-throughput live-cell imaging analysis system includes a base plate and a sample stage above it. A first platform is slidably mounted on the top of the base plate along the X direction and is driven to slide by a first lead screw motor. A second platform is slidably mounted on the first platform along the Y direction and is driven to slide by a second lead screw motor. A vertical slider is slidably mounted on the top of the second platform along the Z direction and is driven to slide by a precision lead screw motor. A fluorescence imaging mechanism is mounted on the vertical slider.

[0007] A gantry-shaped cantilever is provided on the base plate along the Y direction. One end of the gantry-shaped cantilever slides through the sample stage. A third platform is provided on the top of the gantry-shaped cantilever along the Y direction. The third platform is driven to slide by a third lead screw motor. A phase difference imaging mechanism is provided on the third platform.

[0008] Preferably, the sample stage is provided with a sample container adapter, and the sample container is disposed inside the sample container adapter.

[0009] Preferably, the fluorescence imaging mechanism includes an optical imaging component and a four-color LED excitation light source mounted on a vertical slider. The optical imaging component includes a tube lens, an electro-optical microscope turntable, and a CMOS camera.

[0010] Preferably, the phase difference imaging mechanism includes a phase difference ring disposed on one side of the third platform. The phase difference ring is slidably disposed on the third platform along the Y direction. The phase difference ring is driven to slide by a fourth lead screw motor. A lens group and a bright field light source are sequentially disposed on the phase difference ring.

[0011] Preferably, the base plate is provided with a first slide rail on both sides, and the bottom of both ends of the first platform is respectively engaged and slidably connected with the two first slide rails. The first platform is provided with a second slide rail on both ends, and the bottom of both ends of the second platform is respectively engaged and slidably connected with the two second slide rails.

[0012] Preferably, the top of the gantry cantilever is provided with a No. 3 slide rail, the bottom of the No. 3 platform is engaged and slidably connected with the No. 3 slide rail, the top of the No. 3 platform is provided with a No. 4 slide rail, and the bottom of the phase difference ring is engaged and slidably connected with the No. 4 slide rail.

[0013] Preferably, a sliding groove is provided on the sample stage, and one end of the gantry-shaped cantilever is located on the side of the sample stage, while the other end slides through the sliding groove.

[0014] Preferably, the base plate and the sample stage are connected by support legs to form a frame structure, and a circuit board is provided on the frame structure.

[0015] In summary, due to the adoption of the above technical solution, the beneficial effects of this utility model are:

[0016] In this invention, the sample stage is fixed, and a motor drives the light source and objective lens to scan, photograph, and analyze multiple cell culture containers. The entire instrument can be placed in a CO2 incubator, allowing cells to achieve long-term live-cell imaging under suitable culture conditions. Each cell culture container can be independently set with different experimental conditions, enabling multiple experiments to be conducted simultaneously in the same environment. Furthermore, the motorized phase contrast switching device, combined with a bright-field illumination source, can achieve high-resolution phase contrast imaging. The integrated excitation source can provide four wavelengths of excitation light to achieve fluorescence imaging of different colors. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the overall structure of this utility model;

[0018] Figure 2 This is the front view of the present invention;

[0019] Figure 3 This is the left view of the present invention;

[0020] Figure 4 This is the right view of the present invention;

[0021] Figure 5 This is a top view of the present invention;

[0022] In the diagram: 1. Base plate; 2. Slide rail 1; 3. Lead screw motor 1; 4. Platform 1; 5. Slide rail 2; 6. Lead screw motor 2; 7. Platform 2; 8. Four-color LED excitation light source; 9. Electro-optical microscope turntable; 10. Tube lens; 11. CMOS camera; 12. Optical imaging components; 13. Vertical slider; 14. Precision lead screw motor; 15. Sample stage; 16. Sample container adapter; 17. Gantry cantilever; 18. Sliding groove; 19. Slide rail 3; 20. Lead screw motor 3; 21. Platform 3; 22. Slide rail 4; 23. Lead screw motor 4; 24. Phase difference ring; 25. Bright field light source; 26. Lens group; 27. Circuit board. Detailed Implementation

[0023] The specific embodiments of this utility model are described in detail below.

[0024] The "range" disclosed in this utility model is defined by a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, which define the boundaries of a particular range. Ranges defined in this way can include or exclude endpoints and can be arbitrarily combined; that is, any lower limit can be combined with any upper limit to form a range. For example, if a range of 10–50 is listed for a specific parameter, it is also expected that ranges of 10–40 and 20–50 are also included. Furthermore, if the minimum range values ​​are listed as 1 and 2, and the maximum range values ​​are listed as 3, 4, and 5, then the following ranges are all expected: 1–3, 1–4, 1–5, 2–3, 2–4, and 2–5. In this application, unless otherwise stated, the numerical range "a–b" represents a shortened representation of any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0–5" means that all real numbers between "0–5" have been listed herein; "0–5" is merely a shortened representation of these numerical combinations.

[0025] Unless otherwise specified, all embodiments and optional embodiments of this application can be combined to form new technical solutions.

[0026] Unless otherwise specified, all technical features and optional technical features of this application may be combined to form new technical solutions.

[0027] Unless otherwise specified, all steps in this application may be performed sequentially or randomly, preferably sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the mention that the method may also include step (c) indicates that step (c) may be added to the method in any order. For example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0028] Unless otherwise specified, the terms "comprising" and "including" as used in this application can be open-ended or closed-ended. For example, "comprising" and "including" can mean that other components not listed may also be included, or that only the listed components may be included.

[0029] Unless otherwise specified, the reaction will proceed under normal temperature and pressure conditions.

[0030] Unless otherwise specified, all parts or percentages are by weight or by weight percentage.

[0031] In this invention, all the substances used are known substances that can be purchased or synthesized by known methods.

[0032] In this invention, all the devices or equipment used are conventional devices or equipment known in the art and are readily available.

[0033] To make the objectives, technical solutions, and advantages of this utility model clearer, the present utility model will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain this utility model and are not intended to limit this utility model.

[0034] Example:

[0035] A high-throughput live-cell imaging analysis system, such as Figure 1-5 As shown, the sample includes a base plate 1 and a sample stage 15 above it. A first platform 4 is slidably mounted on the top of the base plate 1 along the X direction. The first platform 4 is driven to slide by a first lead screw motor 3. A second platform 7 is slidably mounted on the first platform 4 along the Y direction. The second platform 7 is driven to slide by a second lead screw motor 6. A vertical slider 13 is slidably mounted on the top of the second platform 7 along the Z direction. The vertical slider 13 is driven to slide by a precision lead screw motor 14. A fluorescence imaging mechanism is mounted on the vertical slider 13.

[0036] A gantry-shaped cantilever 17 is provided on the base plate 1 along the Y direction. One end of the gantry-shaped cantilever 17 slides through the sample stage 15. A third platform 21 is provided on the top of the gantry-shaped cantilever 17 along the Y direction. The third platform 21 is driven to slide by a third lead screw motor 20. A phase difference imaging mechanism is provided on the third platform 21.

[0037] In one possible embodiment, the sample stage 15 is provided with a sample container adapter 16, and the sample container is disposed in the sample container adapter 16.

[0038] In one possible embodiment, the fluorescence imaging mechanism includes an optical imaging component 12 and a four-color LED excitation light source 8 disposed on a vertical slider 13. The optical imaging component 12 includes a tube lens 10, an electro-optical microscope turntable 9, and a CMOS camera 11.

[0039] In one possible embodiment, the phase contrast imaging mechanism includes a phase contrast ring 24 disposed on one side of the third platform 21. The phase contrast ring 24 is slidably disposed on the third platform 21 along the Y direction. The phase contrast ring 24 is driven to slide by the fourth lead screw motor 23. A lens group 26 and a bright field light source 25 are sequentially disposed on the phase contrast ring 24.

[0040] In one possible embodiment, the base plate 1 is provided with a first slide rail 2 on both sides, the bottom ends of the first platform 4 are respectively engaged and slidably connected with the two first slide rails 2, the bottom ends of the first platform 4 are provided with a second slide rail 5, and the bottom ends of the second platform 7 are respectively engaged and slidably connected with the two second slide rails 5.

[0041] In one possible embodiment, the top of the gantry cantilever 17 is provided with a third slide rail 19, the bottom of the third platform 21 is engaged and slidably connected with the third slide rail 19, the top of the third platform 21 is provided with a fourth slide rail 22, and the bottom of the phase difference ring 24 is engaged and slidably connected with the fourth slide rail 22.

[0042] In one possible embodiment, a sliding groove 18 is provided on the sample stage 15, and one end of the gantry-shaped cantilever 17 is located on the side of the sample stage 15, while the other end slides through the sliding groove 18.

[0043] In one possible embodiment, the base plate 1 and the sample stage 15 are connected by legs to form a frame structure, and a circuit board 27 is provided on the frame structure.

[0044] By adopting the above technical solution:

[0045] Two slide rails 2 and a lead screw motor 3 are fixed on the base plate 1 of the instrument. The platform 4 can slide back and forth on the slide rails 2 with high precision and high repeatability under the drive of the lead screw motor 3 (movement in the Y-axis direction).

[0046] Platform 4 is also fixed with two slide rails 5 and a lead screw motor 6. Platform 7 moves in a high-precision, high-repeatability reciprocating motion (X-axis direction) on slide rails 5 under the drive of lead screw motor 6.

[0047] Platform 7 is equipped with an integrated four-color LED excitation light source 8 and an optical imaging component 12 consisting of a four-position electro-acoustic microscope turntable 9, a tube lens 10, and a research-grade CMOS camera 11. The electro-acoustic microscope turntable is fixed on a vertical slider 13, which is in turn fixed on a precision lead screw motor 14. The precision lead screw motor 14 can be programmed to automatically focus the microscope objectives on the electro-acoustic microscope turntable to achieve optimal imaging results. Depending on experimental requirements, the electro-acoustic microscope turntable 9 can automatically switch between four different microscope objectives, and the excitation light source 8 can also activate different wavelengths of excitation LEDs as needed to achieve clear fluorescence imaging.

[0048] According to different sample containers and different imaging point requirements, the first lead screw motor 3 drives the first platform 4 to realize the precise reciprocating motion of the optical imaging component 12 on the X-axis, the second lead screw motor 6 drives the second platform 7 to realize the precise reciprocating motion of the optical imaging component 12 on the Y-axis, and the precision lead screw motor 14 drives the vertical slider 13 to realize the precise focusing motion on the Z-axis.

[0049] The open sample stage 15 can accommodate up to 6 sample containers at the same time via the sample container adapter 16, including cell culture plates, dishes and culture flasks, etc.

[0050] The gantry-type cantilever 17 is fixed at both ends of the first platform 4, with one end on the outside of the sample stage 15 and the other end passing through the sliding groove 18 on the sample stage 15, and can reciprocate along the sliding groove 18. The gantry-type cantilever 17 moves synchronously with the first platform 4 under the drive of the first motor 3.

[0051] The gantry-type cantilever 17 is fixed with a No. 3 slide rail 19 and a No. 3 lead screw motor 20. The No. 3 platform 21 can reciprocate along the No. 3 slide rail 19 under the drive of the No. 3 lead screw motor 20.

[0052] A bright-field light source 25 and a lens group 26 are mounted on the outer side of the third platform 21, which can provide bright-field imaging. A fourth slide rail 22 and a fourth motor 23 are fixed on the third platform 21. The phase difference ring 24 can reciprocate along the fourth slide rail 22 under the drive of the motor 23. When the phase difference ring 24 moves between the bright-field light source 25 and the lens group 26, it can provide phase difference imaging.

[0053] Through a precise control algorithm, the imaging objective on the electro-optical mirror turntable on platform 7 and the bright field light source 25 on the gantry cantilever 17 are moved synchronously, ensuring that the imaging objective and the illumination source are synchronized and collimated, thus ensuring clear imaging.

[0054] Circuit board 26 is fixed at the front of the instrument and is connected to all motors and light sources via wire number transmission lines. It controls the movement of the motors and the opening and closing of the light sources according to the instructions of the operating system.

[0055] All program controls are performed through a navigation-style operating system installed on the computer, enabling automatic shooting.

[0056] The captured images are analyzed and processed using the applicant's self-developed image algorithm to meet various application analysis requirements of the instrument design.

[0057] In summary, this application discloses an ultra-high throughput live-cell imaging analysis system, including a top beam and multiple slide rails, a bright-field light source, an electric phase-contrast switching ring and drive motors, a sample stage that can accommodate up to six cell culture containers, a multi-position (at least four-position) electro-acoustic microscope turntable, a telescope, a camera, an integrated four-color fluorescence excitation light source, X, Y, and Z precision motion axes and three drive motors, a base, and a fan. Considering that the entire machine needs to be placed in a CO2 incubator and withstand high temperature and humidity environments, this design incorporates meticulous tri-proof design in the control system, overall machine layout, and ventilation. The self-developed image analysis algorithm can accurately analyze the captured sample images.

[0058] This application allows for real-time imaging of live cells in up to six or more microplates / dishes / flasks under computer control. The software automatically analyzes the images, providing experimental data and results, achieving ultra-high-throughput cell analysis. This significantly improves cell analysis efficiency and data accuracy. It can image multiple cell culture vessels with the same parameters or set individual imaging parameters for each vessel. The motorized multi-position objective turret automatically switches microscope objectives according to preset programs, enabling multi-magnification cell image analysis. An integrated multi-color fluorescence excitation source allows for simultaneous excitation analysis of different fluorescent labels without the need for filter switching.

[0059] The above are merely preferred embodiments of the present utility model and are not intended to limit the present utility model. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present utility model should be included within the protection scope of the present utility model.

Claims

1. A high-throughput live-cell imaging analysis system, characterized in that: Includes a base plate (1) and a sample stage (15) above it. The top of the base plate (1) is provided with a first platform (4) that slides along the X direction. The first platform (4) is driven to slide by a first lead screw motor (3). The first platform (4) is provided with a second platform (7) that slides along the Y direction. The second platform (7) is driven to slide by a second lead screw motor (6). The top of the second platform (7) is provided with a vertical slider (13) that slides along the Z direction. The vertical slider (13) is driven to slide by a precision lead screw motor (14). The vertical slider (13) is provided with a fluorescence imaging mechanism. A gantry-shaped cantilever (17) is provided on the base plate (1) along the Y direction. One end of the gantry-shaped cantilever (17) slides through the sample stage (15). A third platform (21) is provided on the top of the gantry-shaped cantilever (17) along the Y direction. The third platform (21) is driven to slide by a third lead screw motor (20). A phase difference imaging mechanism is provided on the third platform (21).

2. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The sample stage (15) is provided with a sample container adapter (16), and the sample container is located inside the sample container adapter (16).

3. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The fluorescence imaging mechanism includes an optical imaging component (12) mounted on a vertical slider (13) and a four-color LED excitation light source (8). The optical imaging component (12) includes a tube lens (10), an electro-optical microscope turntable (9), and a CMOS camera (11).

4. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The phase difference imaging mechanism includes a phase difference ring (24) disposed on one side of the third platform (21). The phase difference ring (24) is slidably disposed on the third platform (21) along the Y direction. The phase difference ring (24) is driven to slide by the fourth lead screw motor (23). A lens group (26) and a bright field light source (25) are sequentially disposed on the phase difference ring (24).

5. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The base plate (1) is provided with a first slide rail (2) on both sides. The bottom of the first platform (4) is engaged and slidably connected with the two first slide rails (2) respectively. The first platform (4) is provided with a second slide rail (5) on both sides. The bottom of the second platform (7) is engaged and slidably connected with the two second slide rails (5) respectively.

6. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The top of the gantry cantilever (17) is provided with a No. 3 slide rail (19), the bottom of the No. 3 platform (21) is engaged and slidably connected with the No. 3 slide rail (19), the top of the No. 3 platform (21) is provided with a No. 4 slide rail (22), and the bottom of the phase difference ring (24) is engaged and slidably connected with the No. 4 slide rail (22).

7. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The sample stage (15) is provided with a sliding groove (18), and one end of the gantry cantilever (17) is located on the side of the sample stage (15), while the other end slides through the sliding groove (18).

8. The high-throughput live-cell imaging analysis system as described in claim 1, characterized in that: The base plate (1) and the sample stage (15) are connected by support legs to form a frame structure, and a circuit board (27) is provided on the frame structure.

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