Boron-carrying self-luminous microorganism neutron capture bioeffect detection sensor and method

Abstract

This invention discloses a sensor and method for detecting the neutron capture biological effect of boron-bearing self-luminescent microorganisms, belonging to the field of radiation detection technology. It includes self-luminescent microorganisms, a boron-containing component, and a carrier matrix for immobilizing the self-luminescent microorganisms, wherein the boron-containing component includes... 10 Boron compounds of B and a boron-free control group were separated. (Carrying...) 10 B's self-luminescent microorganisms under neutron irradiation 10 B (n,α) 7 The Li reaction produces localized high-linear energy transfer damage, causing changes in luminescence intensity, luminescence ratio, survival rate, or stress response signals. By... 10 Differential analysis between boron-loaded microorganisms and boron-free microorganisms can eliminate background interferences such as gamma rays, fast neutrons, temperature, oxygen, and microbial state, thereby obtaining the boron-dependent neutron capture biological effect signal. This invention can be used for beam quality control in boron neutron capture therapy, biodosage imaging in tissue equivalence models, boron drug screening, and evaluation of neutron field spatial distribution.

Description

Technical Field

[0001] This invention belongs to the field of radiation detection technology, specifically relating to a sensor and method for detecting the biological effects of neutron capture in boron-bearing self-luminescent microorganisms. Background Technology

[0002] Boron neutron capture therapy, BNCT, is a form of addiction. 10 B(n,α) 7 Li nuclear reactions enable precise radiotherapy that achieves localized, highly linear energy transfer damage. The therapeutic effect depends on the coupling relationship between the neutron field, boron distribution, and local biological damage. Existing BNCT dose assessment methods mainly include gold foil activation, thermoluminescent dosimeters, CR-39, scintillation detectors, and Monte Carlo simulations. These methods can be used to measure neutron flux, gamma dose, or physical dose distribution, and are important tools for BNCT beam calibration and dose calculation. However, traditional physical detectors primarily reflect physical radiation field information and are difficult to directly report. 10 Boron participates in the biological effects produced in the local microenvironment after neutron capture. Especially in the mixed radiation field of BNCT, where thermal neutrons, ultrathermal neutrons, fast neutrons, and gamma rays coexist, the neutron flux or absorbed dose alone cannot fully reflect the boron-dependent local high LET biological damage. Self-luminescent microorganisms can continuously generate luminescent signals that can be directly read by optical systems. If... 10 B was introduced into self-luminous microorganisms, and after neutron irradiation... 10 B(n,α) 7 The alpha particles and lithium nuclei produced by the Li reaction can cause localized damage at the microbial scale, leading to changes in luminescence intensity, survival rate, or stress signals. Therefore, self-luminescent microorganisms can serve as living biological transducers, converting boron-dependent neutron capture biological effects into detectable optical signals. However, when using only a single boron-carrying luminescent microorganism for detection, luminescence changes may be simultaneously influenced by factors such as gamma rays, fast neutrons, temperature, oxygen, nutrient status, and microbial growth status, making it difficult to distinguish the true luminescence caused by these factors. 10 The specific response induced by the boron neutron capture reaction. Therefore, it is necessary to develop a boron-carrying self-luminescent microbial sensor with differential correction capabilities, imaging capabilities, and semi-quantitative analysis capabilities. Summary of the Invention

[0003] To address the technical problems described in the background section, the present invention provides a boron-carrying self-luminescent microbial neutron capture biological effect detection sensor and method.

[0004] The technical solution of the present invention is as follows:

[0005] A boron-carrying self-luminescent microbial neutron capture bioeffect detection sensor, comprising:

[0006] Self-luminescent microorganisms;

[0007] Boron-containing components, said boron-containing components include at least those containing... 10 Boron compounds of B;

[0008] A carrier matrix used to immobilize or support the self-luminous microorganisms;

[0009] Wherein, the self-luminescent microorganisms are loaded with the contents of 10 After the boron compound B is used, it produces changes in luminescence signal under neutron irradiation conditions, which can be used to characterize boron-dependent neutron capture biological effects.

[0010] In the above technical solution, the self-luminous microorganisms include naturally luminescent bacteria, engineered self-luminescent bacteria, or microorganisms expressing luminescent reporter systems.

[0011] In the above technical solution, the self-luminescent microorganisms include Vibrio, luminescent bacillus, Shewanella, engineered bacteria expressing the LuxCDABE luminescent operon, or any combination thereof.

[0012] In the above technical solution, the containing 10 The boron compounds of B include one or more of the following: boric acid, borate, boron phenylalanine, sodium mercaptoborane, carborane, boron cluster compounds, boron nanomaterials, boron-containing polymers, or boron-containing biomolecule conjugates.

[0013] In the above technical solution, the sensor further includes a reference detection unit, which includes a load. 11 B. Self-luminescent microorganisms or self-luminescent microorganisms without boron loading.

[0014] In the above technical solution, the load 10 The self-luminescent microorganisms of B and the reference detection unit are located in the same carrier matrix, forming a strip, array, dot matrix, thin film, gel layer or multilayer structure.

[0015] In the above technical solution, the carrier matrix includes one or more of the following: agarose, alginate, gelatin, collagen, polyvinyl alcohol, hydrogel, polyethylene glycol hydrogel, cellulose material, porous membrane, microfluidic chip, paper-based material or tissue equivalent material.

[0016] In the above technical solution, the change in the luminous signal includes a decrease in luminous intensity, an increase in luminous intensity, a change in the luminous ratio, a change in luminous emission in response to stress, a change in the spatial distribution of luminous emission, or a combination thereof.

[0017] A method for preparing the above-mentioned sensor includes the following steps:

[0018] Cultivating self-luminous microorganisms;

[0019] The self-luminescent microorganisms and containing 10 Incubation with boron compounds of B allows the self-luminous microorganisms to be loaded. 10 B;

[0020] Load 10 B's self-luminescent microorganisms are immobilized in a carrier matrix;

[0021] Preparation containing load 10 The sensor in the detection area and the reference detection area of ​​B.

[0022] In the above technical solution, the reference detection area is composed of a load. 11 B consists of self-luminescent microorganisms or self-luminescent microorganisms without boron loading.

[0023] A method for detecting boron-dependent neutron capture biological effects using the aforementioned sensor includes the following steps:

[0024] Acquire the initial emission image or emission intensity of the sensor before neutron irradiation;

[0025] The sensor is placed in the neutron field to be detected for irradiation;

[0026] To obtain images of the emitted light or the intensity of the light after irradiation;

[0027] The biological response value of neutron capture was calculated based on the changes in the emission signal before and after irradiation.

[0028] In the above technical solution, the response value is calculated according to the following formula: R = -ln(Iafter / Ibefore), where Ibefore is the luminous intensity before irradiation and Iafter is the luminous intensity after irradiation.

[0029] In the above technical solution, the boron-dependent neutron capture differential response is obtained through the following formula: R_BNC = R_10B - R_ref. Where R_10B is the response value of the detection region loaded with 10B, and R_ref is the response value of the reference region loaded with 11B or the boron-free reference region.

[0030] In the above technical solution, the neutron field includes a thermal neutron field, an ultrathermal neutron field, a boron neutron capture therapy beam field, a mixed neutron-γ ray radiation field, or a neutron field modulated by a tissue equivalent model.

[0031] The above sensors can be used for any of the following non-therapeutic purposes:

[0032] Boron neutron capture therapy beam quality control;

[0033] Imagery of the spatial distribution of neutron irradiation field;

[0034] Evaluation of biological dose distribution in tissue equivalence models;

[0035] Screening for neutron capture effects in boron drugs or boron-containing materials;

[0036] Evaluation of boron-dependent biological effects in mixed radiation fields;

[0037] Evaluation of thermal neutron shielding materials or neutron modulation materials.

[0038] Beneficial effects:

[0039] Compared with the prior art, the present invention has the following advantages:

[0040] 1. Reportable boron-dependent biological effects:

[0041] This invention does not simply measure neutron flux, but utilizes boron-carrying living microorganisms as reporters. 10 B participates in the local biological effects following neutron capture reactions.

[0042] 2. Possesses differential correction capability:

[0043] pass 10 B / 11 B or 10 B / boron-free differential design can reduce non-specific background interference caused by gamma rays, fast neutrons and environmental factors.

[0044] 3. Enables two-dimensional optical imaging:

[0045] By preparing self-luminescent microorganisms into films, gels, or arrays, spatial distribution maps of neutron capture biological effects can be obtained.

[0046] 4. Applicable to BNCT scenarios:

[0047] It can be used for BNCT beam quality control, phantom validation, boron drug screening, and evaluation of boron-dependent effects in mixed radiation fields.

[0048] 5. Simple detection method:

[0049] The luminescent signal can be read by ordinary CCD, CMOS, gel imaging system or live imaging system without the need for complex electrical detection structure. Attached Figure Description

[0050] Figure 1 This is a schematic diagram of the structure of the sensor of the present invention, including... 10 B. Detection area and reference area.

[0051] Figure 2 This is a photograph of the actual sensor of this invention. The left side shows the sensor being carried. 10The luminescent bacteria in group B are shown on the right, while the luminescent bacteria in the unmodified group are shown on the right.

[0052] Figure 3 This is a schematic diagram illustrating the detection principle of the present invention.

[0053] Figure 4 This invention is based on 10 B(n,α) 7 Li reaction induces changes in the luminescence signal of autoluminescent microorganisms.

[0054] Figure 5 This is the imaging result of the present invention for two-dimensional neutron capture biological effects imaging. Detailed Implementation

[0055] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. However, the following embodiments are only for explaining the present invention, and the scope of protection of the present invention should include all the contents of the claims. Moreover, through the description of the following embodiments, those skilled in the art can fully implement all the contents of the claims of the present invention.

[0056] Example 1: Preparation of a boron-carrying self-luminous microbial sensing unit;

[0057] Stable, self-luminescent microorganisms were selected as the sensing unit. For example... Figure 1 As shown, the self-luminous microorganism can be a naturally occurring self-luminous bacterium or an engineered bacterium expressing the luxCDABE luminescent system.

[0058] The luminescent microorganisms were inoculated into a suitable culture medium and cultured to the logarithmic growth phase. After collecting the cells, they were washed with buffer and then added with... 10 B was incubated with a boron compound and a boron-free buffer solution.

[0059] After incubation, washing removes unbound or untaken boron compounds to obtain the loaded product. 10 B. Self-luminescent microorganisms and boron-free reference microorganisms. For example... Figure 2 As shown, the left side is carrying 10 The luminescent bacteria in group B are shown on the right, while those in the unmodified group are shown on the right. The boron content in the microorganisms was determined using ICP-OES, and the baseline luminescence intensity was measured using a luminescence imaging system. The results showed that a boron content of 2 mg / g had no significant effect on the bacterial luminescence value.

[0060] Example 2: Preparation of differential luminescent microbial films;

[0061] Load 10 B's self-luminescent microorganisms are mixed with the hydrogel precursor solution to form 10 Detection area B. Boron-free luminescent microorganisms were mixed with the same hydrogel precursor solution to form a reference detection area. Samples such as... Figure 2As shown.

[0062] By fabricating two or more of the above-mentioned regions on the same substrate, stripe, dot matrix, checkerboard, microarray, or multilayer thin film structures can be formed. For example... Figure 3 As shown, express 10 The luminescence intensity of detection area B after irradiation express 10 Initial luminescence intensity of detection area B Indicates the luminescence intensity after irradiation of the reference area. Indicating the initial luminescence intensity of the reference region, the detection principle of this invention is as follows: carrying 10 B's self-luminescent microorganisms under neutron irradiation 10 B(n,α) 7 The Li reaction produces localized high LET damage, leading to changes in the luminescence signal. The boron-dependent neutron capture biological effect response is obtained through differential calculation with a reference region. The hydrogel matrix can be agarose, sodium alginate, gelatin, polyvinyl alcohol, polyethylene glycol hydrogel, or other biocompatible matrices. After preparation, the film is subjected to luminescence imaging to obtain the initial luminescence image before irradiation.

[0063] Example 3: Neutron irradiation response detection;

[0064] The sensor prepared in Example 2 was irradiated in the neutron field to be detected with an irradiation flux of 5 × 10⁻⁶. 7 n / cm 2 / s.

[0065] The luminescence intensity was recorded before and after irradiation. The results are as follows: Figure 4 As shown, the total luminescence decay response rate was 65.79% according to the formula, of which the BNCT-specific luminescence decay response rate was 49.20%, and the luminescence decay response rate of other effects (non-boron-mediated background killing) was 16.59%. This indicates that the system can effectively quantify the boron-dependent neutron capture biological effects.

[0066] Example 4: Two-dimensional imaging of biological effects of neutron capture;

[0067] A two-dimensional thin film containing boron-carrying luminescent microorganisms was prepared and placed within a neutron irradiation region. A neutron beam with a 20 cm thick boron-containing polyethylene shielding region was placed above the film, and luminescence images were acquired after irradiation. The results are as follows: Figure 5 As shown, there is no significant difference in light intensity between the shaded and unshaded areas when boron modification is not performed. 10 The luminescent bacterial film region modified with B shows that the shaded area is significantly brighter than the unshaded area, indicating that the film of the present invention can be used to detect two-dimensional neutron capture biological effects and to evaluate the neutron beam quality in the treatment room.

[0068] The above description is merely a specific embodiment of this application, enabling those skilled in the art to understand or implement this application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this application. Therefore, this application is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A boron-carrying self-luminescent microbial neutron capture bio-effect detection sensor, characterized in that, include: Self-luminescent microorganisms; Boron-containing components, said boron-containing components include at least those containing... 10 Boron compounds of B; A carrier matrix used to immobilize or support the self-luminous microorganisms; Wherein, the self-luminescent microorganisms are loaded with the contents of 10 After the boron compound B is used, it produces changes in luminescence signal under neutron irradiation conditions, which can be used to characterize boron-dependent neutron capture biological effects.

2. The sensor according to claim 1, characterized in that, The self-luminescent microorganisms include naturally occurring self-luminescent bacteria, engineered self-luminescent bacteria, or microorganisms expressing luminescent reporter systems.

3. The sensor according to claim 2, characterized in that, The self-luminescent microorganisms include Vibrio, luminescent bacillus, Shewanella, engineered bacteria expressing the LuxCDABE luminescent operon, or any combination thereof.

4. The sensor according to claim 1, characterized in that, The containing 10 The boron compounds of B include one or more of the following: boric acid, borate, boron phenylalanine, sodium mercaptoborane, carborane, boron cluster compounds, boron nanomaterials, boron-containing polymers, or boron-containing biomolecule conjugates.

5. The sensor according to claim 1, characterized in that, The sensor further includes a reference detection unit, which includes a load. 11 B. Self-luminescent microorganisms or self-luminescent microorganisms without boron loading.

6. The sensor according to claim 5, characterized in that, The load 10 The self-luminescent microorganisms of B and the reference detection unit are located in the same carrier matrix, forming a strip, array, dot matrix, thin film, gel layer or multilayer structure.

7. The sensor according to claim 1, characterized in that, The carrier matrix includes one or more of the following: agarose, alginate, gelatin, collagen, polyvinyl alcohol, hydrogel, polyethylene glycol hydrogel, cellulose material, porous membrane, microfluidic chip, paper-based material, or tissue equivalent material.

8. The sensor according to claim 1, characterized in that, The changes in the luminous signal include a decrease in luminous intensity, an increase in luminous intensity, a change in the luminous ratio, a change in luminous response to stress, a change in the spatial distribution of luminous emission, or a combination thereof.

9. A method for preparing the sensor according to any one of claims 1-8, characterized in that, Includes the following steps: Cultivating self-luminous microorganisms; The self-luminescent microorganisms and containing 10 Incubation with boron compounds of B allows the self-luminous microorganisms to be loaded. 10 B; load 10 B's self-luminescent microorganisms are immobilized in a carrier matrix; Preparation containing load 10 The sensor in the detection area and the reference detection area of ​​B.

10. A method for detecting boron-dependent neutron capture biological effects using the sensor described in any one of claims 1–8, characterized in that, Includes the following steps: Acquire the initial emission image or emission intensity of the sensor before neutron irradiation; The sensor is placed in the neutron field to be detected for irradiation; To obtain images of the emitted light or the intensity of the light after irradiation; The biological response value of neutron capture was calculated based on the changes in the emission signal before and after irradiation.

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