Living cell microbeam directional quantitative irradiation imaging device and method

Abstract

The present invention relates to a living cell microbeam directional quantitative irradiation imaging device, belonging to the field of the intersection of radiation biology and photoelectric detection technology. It solves the problem that cell irradiation technology can only perform qualitative irradiation on biological living cells and qualitatively analyze the mechanism of action of biological cells being irradiated, but cannot accurately study the mechanism of action of different irradiation dosages on biological cells. It includes a vertical microbeam terminal, a living cell directional irradiation module, a wide-field microscopy module, a mode switching module and a single proton counting and radiation synchronization control module. The vertical microbeam terminal, the living cell directional irradiation module, the mode switching module and the wide-field microscopy module are coordinated in sequence. The mode switching module is connected to the single proton counting and radiation synchronization control module, and the vertical microbeam terminal is coordinated with the single proton counting and radiation synchronization control module. It realizes accurate quantitative and directional irradiation of biological living cells, thereby better studying the quantitative effect of radiation on cells, and can be used for accurate research on the mechanism of action of different irradiation dosages on biological cells.

Description

Technical Field

[0001] The invention relates to a directional quantitative irradiation device and method, and belongs to the cross-technical field of radiation biology and photoelectric detection. Background Art

[0002] Devices and methods for live-cell microbeam directional quantitative irradiation imaging have important backgrounds and significance in multiple fields. In biology and medicine, the effects of radiation on cells and organisms are a widely studied topic. Studying the mechanisms of radiation action on cells can help us understand fundamental issues in radiation biology, such as DNA repair, cell cycle arrest, and cell death. Radiation is also widely used in medical treatments, such as radiotherapy and radiosensitization therapy. Therefore, devices and methods for live-cell microbeam in situ quantitative irradiation can provide a method for precisely controlling radiation dose and irradiation time, which is of great significance for studying the biological effects of radiation on cells and developing new treatment strategies.

[0003] In the aerospace field, it can be applied in ground-based simulations of space environments to assess the effects of radiation on organisms. Radiation conditions in space can be simulated on the ground to conduct experimental studies on biological materials and radiation protection materials to evaluate their radiation resistance and cytotoxic effects. Furthermore, using live cell microbeam in situ quantitative irradiation devices and methods, the effects of radiation on the cells and tissues of space organisms can be studied, providing fundamental data and scientific basis for solving space biology problems.

[0004] In the field of physics, radiation physics is the study of the interaction of electromagnetic radiation, particle radiation, and nuclear radiation with matter. The interaction between microscopic particle beams, such as ion and electron beams, and matter is a key research area. Using devices and methods for in situ quantitative microbeam irradiation of living cells, we can study the mechanisms of interaction between microscopic particle beams and cells, as well as physical phenomena such as the energy distribution of particle beams deposited in cells, and the scattering and interaction of particles. This is of great significance for understanding fundamental issues in radiation physics and developing new radiation sources and treatments.

[0005] Existing cell irradiation technology can only perform qualitative irradiation on living biological cells and qualitatively analyze the mechanism of action of irradiation on biological cells, but cannot accurately study the mechanism of action of different irradiation doses on biological cells.

[0006] Therefore, there is an urgent need to propose a living cell microbeam directional quantitative irradiation imaging device and method to solve the above technical problems. Summary of the Invention

[0007] The purpose of the present invention is to address the problem that existing cell irradiation technology can only qualitatively irradiate living biological cells and qualitatively analyze the mechanism of action of irradiated biological cells, but cannot accurately study the mechanism of action of different irradiation doses on biological cells. By providing a living cell microbeam directional quantitative irradiation imaging device and method, a brief overview of the present invention is provided below to provide a basic understanding of certain aspects of the present invention. It should be understood that this overview is not an exhaustive overview of the present invention. It is not intended to identify key or important parts of the present invention, nor is it intended to limit the scope of the present invention.

[0008] The technical solution of the present invention:

[0009] A living cell microbeam directional quantitative irradiation imaging device includes a vertical microbeam terminal, a living cell directional irradiation module, a wide-field microscopy module, a mode switching module, and a single proton counting and radiation synchronization control module. The vertical microbeam terminal, the living cell directional irradiation module, the mode switching module, and the wide-field microscopy module are coordinated in sequence, the mode switching module is connected to the single proton counting and radiation synchronization control module, and the vertical microbeam terminal is coordinated with the single proton counting and radiation synchronization control module.

[0010] Preferably, the vertical microbeam terminal comprises a vacuum beam pipe and a vacuum window, and the vacuum beam pipe is provided with a vacuum window.

[0011] Preferably: the living cell directional irradiation module includes a two-dimensional displacement stage, a biological sample tray, a living cell culture dish, a first scintillator and a second scintillator, the biological sample tray is placed on the two-dimensional displacement stage, the first scintillator and the second scintillator are arranged on the biological sample tray, the living cell culture dish is placed on the biological sample tray, the first scintillator or the second scintillator is coaxially arranged with the quasi-vacuum window, and the living cell culture dish is placed above the second scintillator position in the biological sample tray.

[0012] Preferably, the living cell culture dish is a small confocal dish, and the thickness of the bottom of the living cell culture dish is less than 0.17 mm.

[0013] Preferably, the mode switching module includes a single proton counting and collecting device, an electric objective turret and an objective lens. The electric objective turret is provided with the single proton counting and collecting device and the objective lens, and the objective lens is focused on the first scintillator.

[0014] Preferably: the single proton counting and radiation synchronization control module includes a multimode optical fiber jumper, a photon counting detector and a beam switch, the single proton counting collection device is connected to the photon counting detector through the multimode optical fiber jumper, the photon counting detector is connected to the beam switch, and the lower part of the vacuum beam pipe is set in the beam switch.

[0015] The method for live cell microbeam directional quantitative irradiation comprises the following steps:

[0016] Step a: The vacuum window of the vertical microbeam terminal outputs a proton beam, and the first scintillator is moved to the center of the vacuum window of the vertical microbeam terminal by a two-dimensional translation stage;

[0017] Step b: Adjust the axial position of the objective lens so that it is focused on the scintillator surface, adjust the accelerator and vertical microbeam parameters to adjust the beam spot to the center of the field of view, and make the beam spot clearly imaged in the field of view of the wide-field camera, and mark the position of the beam spot;

[0018] Step c: Turn off the beam, move the two-dimensional translation stage to move the second scintillator and the cell culture dish to the microbeam irradiation area, that is, the center of the vertical microbeam terminal vacuum window, turn on the white light source in the widefield microscopy module, and adjust the two-dimensional translation stage to move the cells to be irradiated until they coincide with the center of the beam spot;

[0019] Step d: Turn off the white light source in the widefield microscope module, control the motorized objective turret to align the single proton counting and collection device with the cells to be irradiated, set the number of irradiated protons in the photon counter, and turn on the beam. When the number of monitored irradiated protons reaches a predetermined value, that is, the number of photon pulses counted by the photon counter reaches the set value, its high-speed digital IO port generates a high level and feeds it into the beam switch;

[0020] Step e: After receiving the high-level signal, the beam switch generates a high-voltage bias voltage to deflect the proton beam, shuts off the beam, and completes the microbeam directional quantitative irradiation of living cells.

[0021] The present invention has the following beneficial effects:

[0022] The present invention can achieve precise quantitative and targeted irradiation of living biological cells, thereby enabling better research on the quantitative effects of radiation on cells. It can be used for precise research on the mechanisms of action of different irradiation doses on biological cells, and has important application value and significance in multiple disciplines such as biology, medicine, and physics. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a structural diagram of the living cell microbeam directional quantitative irradiation imaging device.

[0024] In the figure, 1-vacuum beam pipe, 2-vacuum window, 3-two-dimensional translation stage, 4-biological sample tray, 5-living cell culture dish, 6-single proton counting and collection device, 7-objective lens, 8-motorized objective wheel, 9-wide-field microscopy module, 10-multimode fiber optic patch cord, 11-photon counting detector, 12-beam switch, 13-first scintillator, 14-second scintillator. DETAILED DESCRIPTION

[0025] To make the objectives, technical solutions, and advantages of the present invention more clearly apparent, the present invention is described below using specific embodiments shown in the accompanying drawings. However, it should be understood that these descriptions are merely illustrative and are not intended to limit the scope of the present invention. In addition, in the following description, descriptions of well-known structures and technologies are omitted to avoid unnecessary confusion of the concepts of the present invention.

[0026] For the convenience of description, if the words "up", "down", "left" and "right" appear in the present invention, they only indicate that they are consistent with the up, down, left and right directions of the drawings themselves, and do not limit the structure. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or component referred to must have a specific orientation, be constructed and operate in a specific orientation. Therefore, they cannot be understood as limiting the present invention.

[0027] Explanation of terms: The terms "install", "connect", "connect", "fix" and the like in the present invention should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral whole; it can be a mechanical connection or an electrical connection, a direct connection, or an indirect connection through an intermediate medium, an internal connection between two elements, or an interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to the specific circumstances.

[0028] Specific implementation method 1: Combination Figure 1 This embodiment is described. The living cell microbeam directed quantitative irradiation imaging device of this embodiment includes a vertical microbeam terminal, a living cell directional irradiation module, a wide-field microscopy module, a mode switching module and a single proton counting and radiation synchronization control module. The vertical microbeam terminal, the living cell directional irradiation module, the mode switching module and the wide-field microscopy module 9 are coordinated in sequence, the mode switching module is connected to the single proton counting and radiation synchronization control module, and the vertical microbeam terminal is coordinated with the single proton counting and radiation synchronization control module; the present invention can realize accurate quantitative and directed irradiation of biological living cells, so that the quantitative effect of radiation on cells can be better studied, and can be used for accurate research on the mechanism of action of different irradiation dosages on biological cells. It has important application value and significance in multiple disciplines such as biology, medicine and physics.

[0029] Specific implementation method 2: Combination Figure 1 This embodiment describes a living cell microbeam directional quantitative irradiation imaging device. The vertical microbeam terminal includes a vacuum beam pipe 1 and a vacuum window 2. The vacuum beam pipe 1 is provided with a vacuum window 2 to generate a proton beam for irradiating living cells. The vertical microbeam terminal emits the proton beam through the vacuum window. The high-precision two-dimensional translation stage can move the sample tray to align the scintillator 1 13 and the scintillator 2 14 with the vacuum window of the microbeam terminal respectively.

[0030] Specific implementation method three: Combination Figure 1 This embodiment describes a live cell microbeam directional quantitative irradiation imaging device of this embodiment. The live cell directional irradiation module includes a two-dimensional displacement stage 3, a biological sample tray 4, a living cell culture dish 5, a first scintillator 13 and a second scintillator 14. The first scintillator 13 and the second scintillator 14 are both circular. The biological sample tray 4 is placed on the high-precision two-dimensional displacement stage 3. The first scintillator 13 and the second scintillator 14 are arranged on the biological sample tray 4. The living cell culture dish 5 is placed on the biological sample tray 4. The first scintillator 13 and the second scintillator 14 are realized by the two-dimensional displacement stage 3. It is coaxially aligned with the quasi-vacuum window 2, that is, the first scintillator is aligned first to determine the beam spot position, and then the second scintillator is aligned through the two-dimensional translation stage to prepare for cell irradiation. The living cell culture dish 5 is placed above the position of the scintillator 2 14 in the biological sample tray 4. The first scintillator 13 is not set below the living cell culture dish 5, and the living cell culture dish 5 is coaxially arranged with the scintillator 2 14 to align with the target cells; the vertical microbeam terminal emits a proton beam through the vacuum window, and the high-precision two-dimensional translation stage can move the sample tray to align the scintillator 1 13 and the scintillator 2 14 with the vacuum window of the microbeam terminal respectively.

[0031] Specific implementation method four: Combination Figure 1 This embodiment is described. In the living cell microbeam directional quantitative irradiation imaging device of this embodiment, the living cell culture dish 5 is a small confocal dish, and the bottom thickness of the living cell culture dish 5 is less than 0.17 mm.

[0032] Specific implementation method five: Combination Figure 1 This embodiment is described. The living cell microbeam directional quantitative irradiation imaging device of this embodiment has a mode switching module including a single proton counting and collecting device 6, an electric objective wheel 8 and an objective lens 7. The electric objective wheel 8 is provided with the single proton counting and collecting device 6 and the objective lens 7. The objective lens 7 is focused on the first scintillator 13. The objective lens 7 is focused on the first scintillator. The beam spot position is determined only by the first scintillator. The second scintillator does not need to be focused by the objective lens. The second scintillator is used to generate a light pulse detected by the detector when the proton beam passes through. The beam spot generated when the proton beam passes through the scintillator is observed by the wide-field microscope module 9, and the position of the beam spot can be marked. The electric objective wheel 8 can switch the objective lens 7 and the single proton counting and collecting device 6, and switch between the microscope and the single proton counting and collecting device; the wide-field microscope module 9 includes a wide-field camera, a white light source and a tube lens to monitor the beam spot position and size.

[0033] Specific implementation method six: combination Figure 1The present embodiment is described. The living cell microbeam directional quantitative irradiation imaging device of the present embodiment, the single proton counting and radiation synchronization control module includes a multimode optical fiber jumper 10, a photon counting detector 11 and a beam switch 12. The reflective collimator of the single proton counting collection device 6 is connected to the photon counting detector 11 through the multimode optical fiber jumper 10. The photon counting detector 11 is connected to the beam switch 12. The beam switch 12 has two plates. The lower part of the vacuum beam pipe 1 is set between the two plates of the beam switch. The photon counting detector 11 counts the number of set radiation. The number of proton and photon pulses is detected, and its high-speed digital IO port generates a high level and feeds it into the beam switch 12; the single proton counting and collection device 6 includes a coaxially arranged collection lens and a reflective collimator, which collects the photon pulses excited by the proton beam through the scintillator and transmits them to the photon counting detector 11 via a multimode optical fiber jumper 10 with a linear fiber core diameter of 1000μm. After receiving the high-level signal, the beam switch 11 generates a high-voltage bias voltage to deflect the proton beam and turn off the beam; the single proton counting and radiation synchronization control module adopted by the device can realize accurate quantitative irradiation of living cells.

[0034] Specific implementation method seven: combination Figure 1 This embodiment describes a method for directional and quantitative microbeam irradiation of living cells, which uses the aforementioned directional and quantitative microbeam irradiation imaging device for living cells, and includes the following steps:

[0035] Step a: The vacuum window 2 of the vertical microbeam terminal outputs a proton beam, and the first scintillator 13 is moved to the center of the vacuum window 2 of the vertical microbeam terminal via a two-dimensional translation stage 3. The present invention uses a two-dimensional translation stage to accurately move the cells to be irradiated to the center of the beam, achieving precise irradiation at the cellular and subcellular levels.

[0036] Step b: Adjust the axial position of the objective lens 7 so that it is focused on the surface of the first scintillator, adjust the vertical microbeam terminal parameters to adjust the beam spot to the center of the field of view, and make the beam spot clearly imaged in the field of view of the wide-field camera, and mark the position of the beam spot;

[0037] Step c: Turn off the beam, move the two-dimensional translation stage 3 to move the second scintillator 14 and the cell culture dish 5 to the microbeam irradiation area, that is, the center of the vertical microbeam terminal vacuum window 2, turn on the white light source in the wide-field microscope module 9, and fine-tune the two-dimensional translation stage 3 to move the cells to be irradiated in the living cell culture dish to coincide with the center of the beam spot (the location of the marked beam spot); turn the beam on and off using the beam switch, and input a high level to turn off the beam and a low level to turn on the beam from the counting board;

[0038] Step d: Turn off the white light source in the widefield microscope module 9, control the motorized objective turret 8 to align the single proton counting and collecting device 6 with the cells to be irradiated, set the number of irradiated protons in the photon counter 11, and turn on the beam. When the number of monitored irradiated protons reaches a predetermined value, that is, the number of photon pulses counted by the photon counter 11 reaches the set value, the high-speed digital IO port of the photon counter 11 generates a high level and feeds it into the beam switch 12.

[0039] In step e, after receiving the high-level signal, the beam switch 12 generates a high-voltage bias voltage to deflect the proton beam, turns off the beam, and completes the microbeam directional quantitative irradiation of living cells; the radiation dose can be precisely controlled: quantitative irradiation can precisely control the radiation dose of irradiation, so as to better study the quantitative effect of radiation on cells and achieve precise quantitative directional irradiation of living cells; quantitative analysis of cell survival rate: through quantitative irradiation, the cell survival rate can be quantitatively analyzed, so as to better understand the killing effect of radiation on cells; study the response of different cell types: quantitative irradiation can study the response of different cell types, so as to better understand the sensitivity of different cell types to radiation.

[0040] It should be noted that in the above embodiments, as long as the technical solutions are not contradictory, they can be permuted and combined. Those skilled in the art can exhaust all possibilities based on the mathematical knowledge of permutations and combinations. Therefore, the present invention will no longer describe the technical solutions after permutations and combinations one by one, but it should be understood that the technical solutions after permutations and combinations have been disclosed by the present invention.

[0041] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A method for targeted quantitative microbeam irradiation of living cells, characterized by: A living cell microbeam directional quantitative irradiation imaging device is used, comprising a vertical microbeam terminal, a living cell directional irradiation module, a wide-field microscopy module, a mode switching module, and a single proton counting and radiation synchronization control module. The vertical microbeam terminal, the living cell directional irradiation module, the mode switching module, and the wide-field microscopy module (9) are sequentially coordinated, the mode switching module is connected to the single proton counting and radiation synchronization control module, and the vertical microbeam terminal is coordinated with the single proton counting and radiation synchronization control module. The vertical microbeam terminal comprises a vacuum beam pipe (1) and a vacuum window (2), wherein the vacuum beam pipe (1) is provided with the vacuum window (2); The living cell directional irradiation module comprises a two-dimensional displacement stage (3), a biological sample tray (4), a living cell culture dish (5), a first scintillator (13) and a second scintillator (14); the biological sample tray (4) is placed on the two-dimensional displacement stage (3); the first scintillator (13) and the second scintillator (14) are arranged on the biological sample tray (4); the living cell culture dish (5) is placed on the biological sample tray (4); the first scintillator (13) and the second scintillator (14) are coaxially arranged with the vacuum window (2); and the living cell culture dish (5) is placed above the second scintillator (14) in the biological sample tray (4); The mode switching module comprises a single proton counting and collecting device (6), an electric objective lens turret (8), and an objective lens (7); the electric objective lens turret (8) is provided with the single proton counting and collecting device (6) and the objective lens (7); the objective lens (7) is focused on the first scintillator (13); The living cell culture dish (5) is a confocal dish, and the thickness of the bottom of the living cell culture dish (5) is less than 0.17 mm; The single proton counting and radiation synchronization control module comprises a multimode optical fiber jumper (10), a photon counting detector (11) and a beam switch (12); the single proton counting collection device (6) is connected to the photon counting detector (11) via the multimode optical fiber jumper (10); the photon counting detector (11) is connected to the beam switch (12); and the lower part of the vacuum beam pipe (1) is arranged in the beam switch; The method comprises the following steps: Step a: The vacuum window (2) of the vertical microbeam terminal outputs a proton beam, and the first scintillator (13) is moved to the center of the vacuum window (2) of the vertical microbeam terminal via a two-dimensional displacement stage (3); Step b, adjusting the axial position of the objective lens (7) so that it is focused on the surface of the first scintillator, adjusting the accelerator and vertical microbeam parameters to adjust the beam spot to the center of the field of view, and making the beam spot clearly imaged in the field of view of the wide-field camera, and marking the position of the beam spot; Step c, turn off the beam, move the two-dimensional displacement stage (3) to move the second scintillator (14) and the cell culture dish (5) to the microbeam irradiation area, that is, the center of the vertical microbeam terminal vacuum window (2), turn on the white light source in the wide-field microscopy module (9), and adjust the two-dimensional displacement stage (3) to move the cells to be irradiated to coincide with the center of the beam spot; Step d, turning off the white light source in the wide-field microscope module (9), controlling the electric objective lens turret (8) to align the single proton counting and collecting device (6) with the cells to be irradiated, setting the number of irradiated protons for the photon counting detector (11), turning on the beam, and when the number of monitored irradiated protons reaches a predetermined value, that is, the number of photon pulses counted by the photon counting detector (11) reaches a set value, the high-speed digital IO port of the photon counting detector (11) generates a high level and feeds it into the beam switch (12); Step e: After receiving the high-level signal, the beam switch (12) generates a high-voltage bias voltage to deflect the proton beam, shuts down the beam, and completes the microbeam directional quantitative irradiation of living cells.

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