Patent ID: 12259344

In the drawings:1—vacuum beam current pipeline;2—vacuum window;3—two-dimensional displacement table;4—biological sample tray;5—living cell culture dish;6—single-proton counting and collecting apparatus;7—objective lens;8—electronic objective turret;9—wide-field microscopic module;10—multimode fiber patch cord;11—photon counting detector;12—beam current switch;13—first scintillator;14—second scintillator;15—wide-field camera;16—tube lens;17—spectroscope;18—white light source;19—vertical microbeam terminal;20—living cell directional irradiation module;21—mode switching module;22—single-proton counting and radiation synchronous control module.

DETAILED DESCRIPTION OF THE EMBODIMENTS

To make objectives, technical solutions and advantages of the present disclosure more clearly, the present disclosure is described in detail below with reference to specific embodiments shown in the accompanying drawings. It should be understood that these descriptions are only exemplary and are not intended to limit the scope of the present disclosure. In addition, in the following description, the description of well-known structures and technologies are omitted to avoid unnecessarily confusing the concepts of the present disclosure.

For the convenience of illustration, if “up”, “down”, “left”, “right” and the other in the present disclosure only indicate that they are consistent with the up, down, left and right directions of the drawing itself, and are not intended to limit the structure, which are only for convenience of description of the present disclosure and simplification of description rather than indicating or implying that the device or element referred to must have a particular orientation, be constructed and operate in a particular orientation, and thus are not to be construed as limiting the present disclosure.

Terminology explanation part: the terms “install”, “connected”, “connection”, “fix”, and the like should be understood broadly, e.g., a fixed connection, a detachable connection, or an integrated connection; a mechanical connection, or an electrical connection; a direct connection or an indirect connection through an intermediate medium; an internal communication between the two elements or interactions between the two elements. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific situations.

Specific embodiment 1: this embodiment is described in conjunction withFIG.1, a living cell microbeam directional and quantitative irradiation imaging apparatus includes a vertical microbeam terminal, a living cell directional irradiation module, a wide-field microscopic module, a mode switching module, and a single-proton counting and radiation synchronous control module. The vertical microbeam terminal, the living cell directional irradiation module, the mode switching module and the wide-field microscopic module9are sequentially matched, the mode switching module is connected to the single-proton counting and radiation synchronous control module, and the vertical microbeam terminal is matched with the single-proton counting and radiation synchronous control module. Accurate quantitative and directional irradiation of living biological cells can be achieved, thus studying the quantitative influence of radiation on cells better. The apparatus and method can be used to accurately study the mechanism of action of different irradiation doses on biological cells, and have important application values and significance in multiple disciplines such as biology, medicines, and physics.

Specific embodiment 2: this embodiment is described in conjunction withFIG.1, the vertical microbeam terminal of the living cell microbeam directional and quantitative irradiation imaging apparatus includes a vacuum beam current pipeline1, and a vacuum window2. The vacuum beam current pipeline1is provided with the vacuum window2to generate a living cell irradiation proton. The vertical microbeam terminal emits a proton beam current through the vacuum window, the high-precision two-dimensional displacement table can move the sample tray to make a first scintillator13and a second scintillator14aligned with the vacuum window of the microbeam terminal, respectively.

Specific embodiment 3, this embodiment is described in conjunction withFIG.1, the living cell directional irradiation module of the living cell microbeam directional and quantitative irradiation imaging apparatus includes a two-dimensional displacement table3, a biological sample tray4, a living cell culture dish5, a first scintillator13, and a second scintillator14. The first scintillator13and the second scintillator14are both round. The biological sample tray4is placed on the high-precision two-dimensional displacement table3, the first scintillator13and the second scintillator14are arranged on the biological sample tray4, and the living cell culture dish5is placed on the biological sample tray4. The first scintillator13and the second scintillator14are aligned and coaxial with the vacuum window2through the two-dimensional displacement table3, that is, the first scintillator is firstly aligned with the vacuum window to determine a position of a beam spot, and then the second scintillator is aligned with the vacuum window through the two-dimensional displacement table to prepare to irradiate cells. The living cell culture dish5is placed above a position of the second scintillator14in the biological sample tray4, the first scintillator13is not arranged below the living cell culture dish5, and the living cell culture dish5and the second scintillator14are coaxially arranged, such that the second scintillator14is aligned with target cells. The vertical microbeam terminal emits the proton beam current through the vacuum window, and the high-precision two-dimensional displacement table can move the sample tray to make the first scintillator13and the scintillator14aligned with the vacuum window of the microbeam terminal, respectively.

Specific embodiment 4, this embodiment is described in conjunction withFIG.1, the living cell culture dish5of the living cell microbeam directional and quantitative irradiation imaging apparatus is a small confocal dish, and a thickness of the bottom of the living cell culture dish5is less than 0.17 mm.

Specific embodiment 5, this embodiment is described in conjunction withFIG.1, the mode switching module of the living cell microbeam directional and quantitative irradiation imaging apparatus includes a single-proton counting and collecting apparatus6, an electric objective turret8, and an objective lens7. The single-proton counting and collecting apparatus6and the objective lens7are arranged on the electric objective turret8, and the objective lens7is focused to the first scintillator13. When the objective lens7is focused to the first scintillator13, the position of the beam spot can be determined only through the first scintillator, and the second scintillator does not need to be focused by the objective lens, and is used to generate a light pulse to be detected by the detector when the proton beam passes through the second scintillator. The beam spot generated by the proton beam when passing through the scintillator is observed through the wide-field microscopic module9, and a position where the beam spot is located can be marked. The electric objective turret8can switch the objective lens7and the single-proton counting and collecting apparatus6, and switch a microscope and the single-proton counting and collecting apparatus. The wide-field microscopic module9includes a wide-field camera, a white light source, a spectroscope, and a tube lens, and is configured to monitor the position and size of the beam spot.

Specific embodiment 6, this embodiment is described in conjunction withFIG.1, the single-proton counting and radiation synchronous control module of the living cell microbeam directional and quantitative irradiation imaging apparatus includes a multimode fiber patch cord10, a photon counting detector11, and a beam current switch12. A reflective collimator lens of the single-proton counting and collecting apparatus6is connected to the photon counting detector11through the multimode fiber patch cord10, and the photon counting detector11is connected to the beam current switch12. The beam current switch12has two polar plates, a lower portion of the vacuum beam current pipeline1is arranged between the two polar plates of the beam current switch. The photon counting detector11is configured to count the number of photon pulses which has reached a set number of irradiation protons. And a high-speed digital IO port of the photon counting detectors generates a high level to fed to the beam current switch12. The single-proton counting and collecting apparatus6includes a collecting lens and a reflective collimator lens arranged coaxially. The collected photon pulse of the proton excited by the scintillator is transmitted to the photon counting detector11via the multimode fiber patch cord10with a linear core diameter of 1000 μm. The beam current switch12, after receiving a high-level signal, generates a high-voltage bias voltage to deflect the proton beam, and then the beam current is switched off. The single-proton counting and radiation synchronous control module adopted in the apparatus can achieve accurate and quantitative irradiation of living cells.

Specific embodiment 7, this embodiment is described in conjunction withFIG.1, a living cell microbeam directional and quantitative irradiation imaging method employs the of the living cell microbeam directional and quantitative irradiation imaging apparatus, and includes the following steps:Step a. A vacuum window2of a vertical microbeam terminal outputs a proton beam current, a first scintillator13is moved by a two-dimensional displacement table3to the center of the vacuum window2of the vertical microbeam terminal. Through the two-dimensional placement table, cells to be irradiated can be accurately moved to the center of the beam current, thus achieving irradiation at cell and subcellular level.Step b. An axial position where an objective lens7is located is adjusted to make the objective lens7focused to a surface of the first scintillator, and parameters of the vertical microbeam terminal are adjusted to adjust a beam spot to the center of a field of view, such that the beam spot can be clearly imaged in the field of view of the wide-field camera, and a position wherein the beam spot is located is marked.Step c. The beam current is switched off, the two-dimensional displacement table (3) is moved to move a second scintillator14and a cell culture dish5to a microbeam irradiation region, i.e., the center of the vacuum window2of the vertical microbeam terminal. A white light source in a wide-field microscopic module9is switched on, and the two-dimensional displacement table3is finely adjusted to move cells to be irradiated in the living cell culture dish to coincide with a central position of the beam spot (the marked position where the beam spot is located). The beam current is switched on/off through the beam current switch, a counting board feeds a high level to switch off the beam current, and feeds a low level to switch on the beam current.Step d. The white light source in the wide-field microscopic module9is switched off, an electric objective turret8is controlled to make a single-proton counting and collecting apparatus6aligned with the cells to be irradiated, the number of irradiation photons is set for the photon counting detector11, and the beam current is switched on. When monitoring that the number of irradiation protons reaches a predetermined value, that is, the number of photon pulses counted by the photon counting detector11reaches a set value, a high-speed digital IO (input/output) port of the photon counting detector generates a high level, and the high level is fed to a beam current switch12.Step e. The beam current switch12, after receiving a high level signal, generates a high-voltage bias voltage to deflect the proton beam. The beam current is switched off to complete the living cell microbeam directional and quantitative irradiation. The radiation dose can be accurately controlled. The quantitative irradiation can accurately control the radiation dose of irradiation, so as to study the quantitative influence of radiation on cells and achieve accurate, quantitative and directional irradiation on living cells. Quantitative analysis of cell survival rate is as follows: the cell survival rate can be quantitatively analyzed through quantitative irradiation, thus understanding the killing effect of radiation on cells better. The study of the reaction of different cell types is as follows: quantitative irradiation can study the reaction of different cell types, thus understanding the sensitivity of different cell types to radiation better.

It should be noted that in the above embodiments, the technical solutions can be arranged and combined as long as they are not contradictory. Those skilled in the art can exhaust all possibilities according to the mathematical knowledge of arrangement and combination, and thus the technical solutions after arrangement and combination will not be explained one by one in the present disclosure, but it should be understood that the technical solutions after arrangement and combination have been disclosed in the present disclosure.

The above is only the preferred embodiment of the present disclosure, and is not used to limit the present disclosure. For those skilled in the art, the present disclosure may have various modifications and changes. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present disclosure should be included in the scope of protection of the present disclosure.