Weak fluorescence signal acquisition method

Abstract

The invention relates to the technical field of living body fluorescence tomography, and particularly discloses a weak fluorescence signal acquisition method which comprises the following steps: S1, firstly, fixing each structure in an axis limiting groove; then an electric push rod is started to push a center plate to move upwards to the highest position, anesthetic and fluoro-18 deoxyglucose injection is injected into the test mouse to be fixed to the center position of the center plate, and then the positions of a fluorescent CCD, an optical filter, an optical CCD and a laser are changed to be aligned to the test mouse; a side fixing plate, a top plate and a connecting film are all made of light-proof materials and are designed to be in a shading structure, a rotating motor is matched to drive a rotating table, an electric push rod, a guide rod and a center plate to rotate intermittently, a fluorescent CCD and an optical CCD are matched to take 36 pictures in a surrounding mode, all-directional fluorescent signals of a test mouse can be accurately captured, and the detection accuracy is improved. A three-dimensional model is formed by matching with modeling software, so that fluorescence signals on a test mouse body can be captured in all directions, and three-dimensional modeling recording is carried out.

Description

Technical Field

[0001] This invention relates to the field of in vivo fluorescence tomography technology, specifically a method for acquiring weak fluorescence signals. Background Technology

[0002] In biofluorescence tomography, the acquisition of weak fluorescence signals is a crucial step. Only by accurately acquiring weak fluorescence data can three-dimensional models be fluorescently labeled, thereby enabling direct monitoring of cellular activities and gene behavior in living organisms. Through this in vivo imaging system, biological processes such as tumor growth and metastasis, the development of infectious diseases, and the expression of specific genes in living animals can be observed.

[0003] Chinese invention patent CN116626000A, published on August 22, 2023, discloses a microlens-based fluorescence imaging device. The device includes an expandable stage, a micro-dark chamber, an imaging cylinder, and an incident light source. The micro-dark chamber has a hole at its top center and a light-shielding cover movably mounted on its top. It also includes a cylindrical tube, microlenses, and a filter, an image sensor encapsulation cover, and an image sensor photosensitive array arranged sequentially from the bottom of the cylindrical tube downwards. This invention solves the problem of low imaging resolution in traditional lensless fluorescence imaging systems. The invention also discloses a fluorescence image acquisition method. Using the fluorescence imaging device of this invention, the method includes sequential fluorescence processing, turning on the incident light source, and adjusting the position of the microlens to acquire the fluorescence image. This invention achieves efficient fluorescence image acquisition.

[0004] However, the aforementioned publicly available solutions have the following shortcomings: the existing fluorescence acquisition devices have a single acquisition angle, resulting in low acquisition accuracy. They cannot cross-verify fluorescence images from multiple angles, leading to low detection accuracy for weak fluorescence and low overall detection performance. Summary of the Invention

[0005] The purpose of this invention is to solve the above-mentioned technical problems and provide a method for acquiring weak fluorescence signals.

[0006] To achieve the above objectives, the present invention employs the following technical solutions;

[0007] The technical solution adopted by this invention to solve its technical problem is: a weak fluorescence signal acquisition method according to this invention, which includes the following steps:

[0008] S1: First, fix each structure in the axis limiting groove; then start the electric push rod to push the center plate upward to the highest position, then inject the test mouse with anesthesia and fix it in the center position of the center plate, then change the position of the fluorescent CCD, filter, optical CCD and laser to align with the internal organs of the test mouse.

[0009] S2: Insert the top plate into the lifting shaft, rotate the side movable plate to close the top plate and the side fixed plate to form a dark chamber, thereby reducing the influence of visible light on fluorescence collection and improving the accuracy of fluorescence collection;

[0010] S3: Connect the fluorescent CCD, optical CCD, and laser to the computer using a data cable, start the fluorescent CCD, optical CCD, laser, and rotary motor. The rotary motor rotates intermittently 36 times, pausing for two seconds each time. Within the two seconds, the optical CCD takes one picture first, and the fluorescent CCD takes one picture afterward.

[0011] S4: Using computer software, thirty-six photos taken by a fluorescent CCD are combined to form a fluorescent three-dimensional model. Finite element analysis software is used to perform finite element analysis on the three-dimensional model. Through the Boltzmann particle transport equation, finite element method and modern optimization calculation method, the three-dimensional distribution of fluorescent substances in the organism is reconstructed and quantified. The fluorescence intensity is displayed by color gradation and the fluorescent labeling parameters are also analyzed.

[0012] S5: Using a computer and software, combine the thirty-six photos taken by the optical CCD to form a solid three-dimensional model, and align it with the fluorescent three-dimensional model.

[0013] Furthermore, the axis limiting groove is formed on the upper surface of the middle layer plate; a protective cover is fixedly connected to the lower surface of the middle layer plate; a bottom plate is detachably fixedly connected inside the protective cover; a rotary motor is fixedly connected to the upper surface of the bottom plate; a rotary table is fixedly connected to the output shaft of the rotary motor; an electric actuator and a guide rod are fixedly connected to the upper surface of the rotary table; a center plate is fixedly connected to the upper end of the electric actuator and the guide rod; the center plate is located in a center groove; the center groove is formed inside the middle layer plate.

[0014] Furthermore, multiple support units are inserted into the axis limiting groove; a fluorescent CCD, a filter, an optical CCD, and a laser are fixedly connected to the support unit; the filter is symmetrically located on both sides of the central plate.

[0015] Furthermore, the support unit includes a limiting base and a triple square sleeve; the limiting base is inserted into the axis limiting groove; the triple square sleeve is fixedly connected to the upper end of the limiting base; and bolts are threaded into the side openings of the triple square sleeve.

[0016] Furthermore, a lifting shaft is inserted into the middle layer plate; a top layer plate is inserted into the upper end of the lifting shaft; side movable plates are fixed to both sides of the top layer plate; a connecting membrane is fixed to the outer surface of the side movable plates; and side movable plates are fixed to the connecting membrane.

[0017] Furthermore, the side fixing plate, top plate, and connecting membrane are all made of opaque materials.

[0018] The present invention provides a method for acquiring weak fluorescence signals, which has the following beneficial effects:

[0019] 1. This invention utilizes a light-blocking structural design where the side fixing plates, top plate, and connecting membrane are all made of opaque materials. A rotating motor drives the rotating table, electric push rod, guide rod, and center plate to perform intermittent rotational movements. Combined with a fluorescent CCD and an optical CCD to capture thirty-six images in a 3D pattern, the invention can accurately capture the fluorescence signals of the experimental mice from all directions. This, along with modeling software, creates a three-dimensional model, enabling precise recording of the fluorescence signals on the experimental mice. Attached Figure Description

[0020] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings;

[0021] Figure 1 A schematic diagram of an axonometric view of a weak fluorescence signal acquisition device;

[0022] Figure 2 A schematic diagram of the first internal structure of a weak fluorescence signal acquisition device;

[0023] Figure 3 This is a schematic diagram of the second internal structure of a weak fluorescence signal acquisition device;

[0024] Figure 4 This is a schematic diagram of the third internal structure of a weak fluorescence signal acquisition device;

[0025] Figure 5 This is a partial structural diagram of a weak fluorescence signal acquisition device.

[0026] The following are the labeling instructions in the diagram: 1. Bottom layer plate; 2. Protective cover; 3. Middle layer plate; 4. Center groove; 5. Center plate; 6. Electric actuator; 7. Guide rod; 8. Computer; 9. Axis limiting groove; 10. Lifting shaft; 11. Side fixing plate; 12. Top layer plate; 13. Connecting membrane; 14. Side movable plate; 15. Limiting base; 16. Triple square sleeve; 17. Fluorescent CCD; 18. Filter; 19. Optical CCD; 20. Laser; 21. Rotary stage; 22. Rotary motor. Detailed Implementation

[0027] It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention.

[0028] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0029] It should be noted that in the embodiments of the present invention, all directional indications (such as up-down-left-right-forward-backward...) are only used to explain the relative positional relationship and movement between the components in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indication will also change accordingly. The connection can be a direct connection or an indirect connection.

[0030] Please see Figure 1-5 As shown, a method for acquiring weak fluorescence signals includes the following steps:

[0031] S1: First, fix each structure in the axis limiting groove 9; then start the electric push rod 6 to push the center plate 5 upward to the highest position, then inject the test mouse with anesthesia and fix the fluorine-18 deoxyglucose injection solution in the center position of the center plate 5, then change the position of the fluorescent CCD 17, filter 18, optical CCD 19 and laser 20 to align with the internal organs of the test mouse.

[0032] S2: Insert the top plate 12 into the lifting shaft 10, rotate the side movable plate 14 to close the top plate 12 and the side fixed plate 11 to form a dark chamber, so that the influence of visible light on fluorescence collection is reduced and the accuracy of fluorescence collection is improved.

[0033] S3: Connect the fluorescent CCD17, optical CCD19 and laser 20 to the computer 8 using a data cable, start the fluorescent CCD17, optical CCD19, laser 20 and rotary motor 22, the rotary motor 22 rotates intermittently thirty-six times, pausing for two seconds each time, within two seconds, the optical CCD19 takes one picture first, and the fluorescent CCD17 takes one picture afterward.

[0034] S4: Using computer 8 and software, thirty-six photos taken by fluorescent CCD17 are combined to form a fluorescent three-dimensional model. Finite element analysis software is used to perform finite element analysis on the three-dimensional model. Through Boltzmann particle transport equation, finite element and modern optimization calculation methods, the three-dimensional distribution of fluorescent substances in the organism is reconstructed and quantified. Fluorescence intensity is displayed by color scale and fluorescent labeling parameters are also analyzed.

[0035] S5: Using computer 8 and software, combine the thirty-six photos taken by the optical CCD19 to form a solid three-dimensional model, and align it with the fluorescent three-dimensional model.

[0036] The axis limiting groove 9 is formed on the upper surface of the middle layer plate 3; a protective cover 2 is fixedly connected to the lower surface of the middle layer plate 3; a bottom plate 1 is detachably fixed inside the protective cover 2; a rotary motor 22 is fixedly connected to the upper surface of the bottom plate 1; a rotary table 21 is fixedly connected to the output shaft of the rotary motor 22; an electric push rod 6 and a guide rod 7 are fixedly connected to the upper surface of the rotary table 21; a center plate 5 is fixedly connected to the upper end of the electric push rod 6 and the guide rod 7; the center plate 5 is located in the center groove 4; the center groove 4 is formed in the middle layer plate 3; during operation, the rotary motor 22 can drive the rotary table 21, the center plate 5, the electric push rod 6, the guide rod 7 and the experimental mouse to rotate precisely, solving the problem of low rotation accuracy in existing systems.

[0037] Multiple support units are inserted into the axial limiting groove 9; a fluorescent CCD 17, a filter 18, an optical CCD 19, and a laser 20 are fixedly connected to the support unit; the filter 18 is symmetrically located on both sides of the central plate 5; during operation, the fluorescent CCD 17, filter 18, optical CCD 19, and laser 20 are supported by the support unit, so that the height of the fluorescent CCD 17, filter 18, optical CCD 19, and laser 20 can be adjusted to align with the test mouse by adjusting the height of the support unit, thereby obtaining the filter 18, fluorescent CCD 17, and laser 20 symmetrically arranged with the test mouse as the center, and the optical CCD 19 arranged rotationally symmetrically, to obtain the best alignment accuracy, so as to accurately capture the weak fluorescence on the test mouse.

[0038] The support unit includes a limiting base 15 and a triple square sleeve 16; the limiting base 15 is inserted into the axis limiting groove 9; the triple square sleeve 16 is fixed to the upper end of the limiting base 15; the triple square sleeve 16 has a side opening with a threaded bolt inserted into it; during operation, the triple square sleeve 16, composed of three square tubes that fit together, allows the height of the upper fixed structure to be adjusted by changing the overlap distance, thereby adapting to the vertical height of the center plate 5, so that the experimental mouse fixed to the center of the upper surface of the center plate 5 can be aligned.

[0039] A lifting shaft 10 is inserted into the middle layer plate 3; a top layer plate 12 is inserted into the upper end of the lifting shaft 10; side movable plates 14 are fixed to both sides of the top layer plate 12; a connecting membrane 13 is fixed to the outer surface of the side movable plates 14; and the side movable plates 14 are fixed to the connecting membrane 13. During operation, a dark chamber is formed by the side fixed plate 11, the top layer plate 12, the connecting membrane 13, and the side movable plates 14, thereby reducing the influence of indoor lighting on the weak fluorescence on the experimental mice collected by the fluorescence CCD 17 and the optical CCD 19, thus improving the accuracy of the collection.

[0040] The side fixing plate 11, top plate 12, connecting membrane 13 and side movable plate 14 are all made of opaque material. During operation, a dark chamber is formed by the side fixing plate 11, top plate 12, connecting membrane 13 and side movable plate 14, which can reduce the influence of indoor light on the weak fluorescence on the experimental mice collected by the fluorescent CCD 17 and optical CCD 19, thereby improving the accuracy of the collection.

[0041] Using the above scheme, when in use, the side fixing plate 11, top plate 12 and connecting film 13 are all made of opaque materials to block light. With the help of the rotary motor 22, the rotary table 21, electric push rod 6, guide rod 7 and center plate 5 are driven to rotate intermittently. With the help of the fluorescent CCD 17 and optical CCD 19, thirty-six photos are taken around the mouse, which can accurately capture the fluorescence signal of the experimental mouse in all directions. With the help of modeling software, a three-dimensional model is formed, so that the fluorescence signal on the experimental mouse can be captured in all directions and recorded in three dimensions, thus realizing the accurate recording of the fluorescence signal on the experimental mouse.

[0042] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0043] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.

Claims

1. A method for collecting a weak fluorescent (highlight) signal, characterized by; The method comprises the following steps: S1: first, fix each structure in the axis limiting groove (9); then, start the electric push rod (6) to push the center plate (5) to move upward to the highest position, then inject anesthetic and fluorine 18 deoxy glucose injection into the test mouse and fix it in the center position of the center plate (5), then change the positions of the fluorescent CCD (17), the optical filter (18), the optical CCD (19) and the laser (20) to align the internal organ position of the test mouse; S2: insert the top plate (12) into the lifting shaft (10), rotate the side movable plate (14) to close the top plate (12) and the side fixed plate (11) to form a darkroom, so that the influence of visible light on fluorescence collection is reduced, and the accuracy of fluorescence collection is improved; S3: connect the fluorescent CCD (17), the optical CCD (19) and the laser (20) by using the data line through the computer (8), start the fluorescent CCD (17), the optical CCD (19), the laser (20) and the rotating motor (22), the rotating motor (22) rotates intermittently thirty-six times, each time stops for two seconds, and in the two seconds, the optical CCD (19) first takes a picture, and the fluorescent CCD (17) takes a picture; S4: use the computer (8) to combine the thirty-six pictures taken by the fluorescent CCD (17) to form a fluorescent three-dimensional model, use finite element analysis software to perform finite element analysis on the three-dimensional model, through the Boltzmann particle transport equation, finite element and modern optimization calculation method, reconstruct the three-dimensional distribution of the fluorescent substance in the biological body and realize quantification, display the fluorescence intensity through the color scale, and the fluorescence labeling parameters; S5: use the computer (8) to combine the thirty-six pictures taken by the optical CCD (19) to form a solid three-dimensional model, and align the fluorescent three-dimensional model.

2. The method of claim 1, wherein: The axis limiting groove (9) is arranged on the upper surface of the middle layer plate (3); the lower surface of the middle layer plate (3) is fixedly connected with the protective cover (2); the protective cover (2) is detachably fixedly connected with the bottom layer plate (1); the upper surface of the bottom layer plate (1) is fixedly connected with the rotating motor (22); the output shaft of the rotating motor (22) is fixedly connected with the rotating table (21); the upper surface of the rotating table (21) is fixedly connected with the electric push rod (6) and the guide rod (7); the upper ends of the electric push rod (6) and the guide rod (7) are fixedly connected with the center plate (5); the center plate (5) is located in the center groove (4); and the center groove (4) is arranged in the middle layer plate (3).

3. The method of claim 2, wherein: A plurality of support units are inserted into the axis limiting groove (9); the support units are fixedly connected with the fluorescent CCD (17), the optical filter (18), the optical CCD (19) and the laser (20); and the optical filter (18) is symmetrically located on both sides of the center plate (5).

4. The method of claim 3, wherein: The support unit comprises a limiting base (15) and a triple square sleeve (16); the limiting base (15) is inserted into the axis limiting groove (9); the upper end of the limiting base (15) is fixedly connected with the triple square sleeve (16); and the side of the triple square sleeve (16) is threadedly connected with the bolt.

5. The method of claim 4, wherein: The middle layer board (3) is inserted with a lifting shaft (10); the upper end of the lifting shaft (10) is inserted with a top layer board (12); the two sides of the top layer board (12) are fixedly connected with side movable boards (14); the outer surface of the side movable boards (14) is fixedly connected with connecting films (13); the connecting films (13) are fixedly connected with the side movable boards (14).

6. The method of claim 5, wherein: The side fixed boards (11), the top layer board (12) and the connecting films (13) are all made of light-proof material.

Images