A cross-scale, multi-mode, visual biological detection device and detection method

Abstract

The application discloses a kind of cross-scale, multi-mode, visual biological detection device and detection method, belong to biological detection technical field, including: X-ray fluorescence imaging module, mechanical spectroscopy adhesion monitoring module, signal integration processing module.X-ray fluorescence imaging module is used for the acquisition, optimization, coding and identification of biological sample blue, green, red and near-infrared color pattern, mechanical spectroscopy adhesion monitoring module is used for real-time, dynamic monitoring and recording to biological sample active adhesion, through the collaborative processing analysis of X-ray optical signal and mechanical spectroscopy signal of the biological sample to be measured, realize the fluorescence visual of biological sample morphology, structure component and the accurate acquisition of biological activity data, for the research and application of cross-scale, multi-mode, visual contrast agent of mechanical spectroscopy and X-ray excited fluorescence imaging provides equipment support, promote its development optimization and conversion application in biomedical field such as inflammation infection, malignant disease, tumor diagnosis.

Description

Technical Field

[0001] This invention belongs to the field of biological detection technology, specifically relating to a cross-scale, multi-modal, and visualized biological detection device, and also to a cross-scale, multi-modal, and visualized biological detection method. Background Technology

[0002] X-ray excitation optical fluorescence imaging technology, as a novel optical imaging technique, is receiving increasing research and attention in the field of biological detection and diagnosis because both the excitation light (X-rays) and the emission light (near-infrared II region fluorescence) can overcome the limitations of the penetration depth of biological samples and reduce background interference from biological autofluorescence.

[0003] Furthermore, the patent applicant invented a contrast agent that, under X-ray excitation, generates fluorescence signals at different wavelengths, including blue, green, red, and near-infrared, thereby obtaining controllable, colorful, and real-time color images of the organism under test. However, this technology can only qualitatively obtain information on the cell morphology, structure, and composition of the organism under test; it cannot accurately extract features or precisely analyze specific values ​​of biological activity.

[0004] To address the aforementioned issues, the technique of obtaining mechanospectroscopy through monitoring organisms is an important means of detecting the activity and composition information of organisms, with enormous application potential. Its basic principle is based on the real-time monitoring of biological activity through the interaction between integrins on the organism's surface and the surface of a needle. Specifically, as activity decreases, its adhesion decreases, and its morphology gradually changes. At this point, the needle bends to varying degrees, and the maximum bending displacement of the needle tip, i.e., the amplitude, can quantitatively and accurately assess the activity data of the biological sample.

[0005] Therefore, by combining the two technologies mentioned above and utilizing their advantages and characteristics, a novel detection device can be designed that integrates bioactivity and composition information obtained by mechanospectroscopy with structural and composition information obtained by X-ray excitation fluorescence imaging. This device, using a self-invented contrast agent material, enables objective, continuous, real-time, and accurate biological detection across scales, multiple modes, and visualization. This represents a major breakthrough in biological detection technology. Summary of the Invention

[0006] This invention addresses the aforementioned problems in existing technologies by proposing a cross-scale, multi-modal, and visualized biological detection device and method that enables fluorescence visualization of biological sample morphology and structural components, as well as accurate acquisition of bioactivity data.

[0007] This invention can be achieved through the following technical solutions:

[0008] A multi-scale, multi-modal, visualized biological detection device, comprising:

[0009] Stage, used to hold biological samples containing contrast agents;

[0010] The X-ray fluorescence imaging module is located on top of the stage. The X-ray fluorescence imaging module includes a fluorescence signal excitation unit, a fluorescence signal processing unit, and an image processing unit. The fluorescence signal excitation unit is used to emit X-rays that can irradiate biological samples. The fluorescence signal processing unit is used to collect and screen the fluorescence signals generated after the X-ray excitation of the contrast agent. The image processing unit is used to receive the screened fluorescence signals and generate color image signals.

[0011] A mechanical spectroscopic adhesion monitoring module includes nanoneedles that can adhere to biological samples, and monitors the active adhesion of biological samples in real time by the degree of bending of the nanoneedles.

[0012] The signal integration and processing module is used to receive, process and analyze signals from the X-ray fluorescence imaging module and the mechanospectral adhesion monitoring module, and to realize fluorescence visualization of the morphology and structural components of biological samples and the acquisition of bioactivity data.

[0013] As a further improvement of the present invention, the fluorescence signal excitation unit includes an X-ray excitation tube that provides an X-ray radiation source for the biological sample. The X-ray excitation tube is positioned toward the biological sample on the stage, and an X-ray collimator is provided at the exit end of the X-ray excitation tube.

[0014] As a further improvement of the present invention, the fluorescence signal processing unit includes:

[0015] An optical objective lens is positioned to be aligned with the biological sample on the stage, and the optical objective lens is used to receive fluorescence signals;

[0016] A filter array, which is detachably installed in the fluorescence signal transmission channel, is used to intercept and filter fluorescence signals of different wavelengths.

[0017] As a further improvement of the present invention, the image processing unit includes an imaging camera disposed at the end of the fluorescence signal transmission channel. The imaging camera is used to receive the fluorescence signal filtered by the filter group and produce a color image signal.

[0018] As a further improvement of the present invention, the filter group includes a monochrome filter group or a color filter array.

[0019] As a further improvement of the present invention, a short-pass dichroic mirror 250 is provided between the filter group and the imaging camera.

[0020] As a further improvement of the present invention, the imaging camera includes a visible light imaging camera 241 and a near-infrared imaging camera 242. The near-infrared light emitted by the biological sample after X-ray radiation is transmitted through the short-pass dichroic mirror 250 and collected by the near-infrared imaging camera 242. The visible light emitted by the biological sample after X-ray radiation is reflected through the short-pass dichroic mirror 250 and collected by the visible light imaging camera 241.

[0021] As a further improvement of the present invention, the mechanospectral adhesion monitoring module also includes a mechanical motion signal acquisition device for detecting and recording the bending amplitude of the nanoneedle.

[0022] As a further improvement of the present invention, the signal integration processing module includes:

[0023] A digital image processing terminal, used to receive and process the color image signals from the imaging camera;

[0024] A mechanical motion signal analysis terminal is used to receive and process the data collected by the mechanical motion signal acquisition device.

[0025] A multi-scale, multi-modal, and visualized biological detection method includes steps S11-S14 and S21-S23, wherein S11-S14 and S21-S23 can be performed simultaneously. The method is characterized by the following specific steps:

[0026] S11. The X-ray excitation tube emits X-rays to irradiate the surface of the biological sample. At this time, the biological sample containing the contrast agent is excited and produces a fluorescence change.

[0027] S12. The optical objective lens recovers the fluorescence signal generated after the biological sample containing the contrast agent is excited, and intercepts and filters the fluorescence signals of different wavelengths through the filter group. The filtered fluorescence signal is transmitted to the imaging camera and generates a color image signal, which is then transmitted to the digital image processing terminal.

[0028] S13. The digital image processing terminal sequentially performs image enhancement and image restoration on the received color image signal to eliminate interference and blurring of image quality, weaken background fluorescence, highlight the morphological structure information of biological cells in the image, and obtain the restored image.

[0029] S14. The digital image processing terminal sequentially performs image encoding and image recognition on the restored image to identify the component information in the biological sample imaging image.

[0030] S21. The nano-needle is immersed in the biological sample by an external microcomputer, and the position of the nano-needle when no biological sample adheres is recorded as the equilibrium point. The position of the equilibrium point is recorded by the mechanical motion signal acquisition device and transmitted to the mechanical motion signal analysis terminal.

[0031] S22. After the nano-needle is cultured in the biological sample for a period of time, the nano-needle bends due to the adhesive behavior of the biological sample. The location where the tip of the nano-needle bends is recorded again by the mechanical motion signal acquisition device and transmitted to the mechanical motion signal analysis terminal.

[0032] S23. The mechanical motion signal acquisition device continuously acquires the bending amplitude of the tip of the nano-needle and transmits the signal to the mechanical motion signal analysis terminal. The mechanical motion signal analysis terminal identifies the amplitude, variation law and waveform distortion characteristics of the signal time-domain waveform to perform quantitative data evaluation and dynamic continuous real-time monitoring of the activity status of the biological sample, and obtains accurate biological sample activity data.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] 1. The X-ray fluorescence imaging module acquires color images of biological samples, while the mechano-spectroscopy adhesion monitoring module records real-time dynamic monitoring of the adhesion of biological samples. The signal integration and processing module then performs collaborative processing and analysis on the color images and the real-time dynamic monitoring records of the adhesion of biological samples, enabling the fluorescence visualization of the morphology and structural components of biological samples and the accurate acquisition of bioactivity data. This constructs a cross-scale, multi-mode, and visualized biological detection device that integrates adhesion mechano-spectroscopy and X-ray excitation fluorescence imaging.

[0035] 2. The innovative combination of fluorescence image information and adhesion mechanical spectroscopy is applied to biological detection, providing equipment support for the research and application of similar biological detection contrast agents, and promoting their development, optimization and translational application in biomedical fields such as inflammation and infection, malignant diseases, and tumor diagnosis.

[0036] 3. Based on a contrast agent with the composition NaAF4:B (A = Y, Zr, Nb, Mo, Tc, Fe, Mn, Bi; B = Ce, Nd, Ho, Er, Tm, Yb), the blue, green, red, and near-infrared color images of biological samples are acquired after X-ray excitation. The color images are then optimized, encoded, and recognized by a digital image processing terminal, successfully obtaining the component information in the imaging images of biological samples.

[0037] 4. Based on the multi-wavelength and tunable emission characteristics of the contrast agent NaAF4:B (A = Y, Zr, Nb, Mo, Tc, Fe, Mn, Bi; B = Ce, Nd, Ho, Er, Tm, Yb) under X-ray excitation, the corresponding filter group is set to provide the function of monochromatic filtering and color filtering, so as to realize the purpose of generating single-channel fluorescence imaging or three-primary-color multicolor imaging of biological samples containing contrast agents after X-ray excitation, such as blue, red, green, and near-infrared.

[0038] 5. The mechanical motion signal acquisition device detects and records the bending amplitude of the nano-suspended needle and transmits it to the mechanical motion signal analysis terminal. The mechanical motion signal analysis terminal can analyze and process the speed of signal change, the range of signal amplitude, and the continuity and discreteness of the signal. By identifying the amplitude, change law, and waveform distortion characteristics of the signal time-domain waveform, it can then perform quantitative data evaluation and dynamic continuous real-time monitoring of the activity status of biological samples. Attached Figure Description

[0039] Figure 1 This is a schematic diagram of the structure of the cross-scale, multi-modal, and visualized biological detection device of the present invention;

[0040] Figure 2 This is a schematic diagram of the process of the cross-scale, multi-modal, and visualized biological detection device of the present invention;

[0041] Figure 3 This is a schematic diagram illustrating the detection theory of the cross-scale, multi-mode, and visualized biological detection device of the present invention.

[0042] In the figure, 100 is the stage; 200 is the X-ray fluorescence imaging module; 210 is the X-ray excitation tube; 211 is the X-ray collimator; 220 is the optical objective lens; 230 is the filter group; 240 is the imaging camera; 241 is the visible light imaging camera; 242 is the near-infrared imaging camera; 250 is the short-pass dichroic mirror; 300 is the mechanical spectroscopic adhesion monitoring module; 310 is the nanoneedle; 320 is the mechanical motion signal acquisition device; 400 is the signal integration and processing module; 410 is the digital image processing terminal; and 420 is the mechanical motion signal analysis terminal. Detailed Implementation

[0043] The following are specific embodiments of the present invention, which are described in conjunction with the accompanying drawings to further illustrate the technical methods of the present invention. However, the present invention is not limited to these embodiments.

[0044] like Figure 1-3 As shown, the present invention provides a cross-scale, multi-modal, and visualized biological detection device, comprising:

[0045] Stage 100, used to place biological samples containing contrast agents;

[0046] The X-ray fluorescence imaging module 200 is located on top of the stage 100. The X-ray fluorescence imaging module 200 includes a fluorescence signal excitation unit, a fluorescence signal processing unit, and an image processing unit. The fluorescence signal excitation unit is used to emit X-rays that can irradiate biological samples. The fluorescence signal processing unit is used to collect and screen the fluorescence signals generated after the X-ray excitation contrast agent. The image processing unit is used to receive the screened fluorescence signals and generate color image signals.

[0047] The mechanospectral adhesion monitoring module 300 includes nanoneedles 310 that can adhere to biological samples. It monitors the active adhesion of biological samples in real time by measuring the bending degree of the nanoneedles 310. It is worth mentioning that the reason why the bending degree of the nanoneedles 310 can be used to monitor the active adhesion of biological samples is based on the interaction between integrins on the surface of the organism and the surface of the nanoneedles 310. Specifically, when the nanoneedles 310 and the biological sample are co-cultured for a certain period of time, as the activity of the biological sample decreases, its adhesion decreases and its morphology gradually changes. At this time, the morphological changes of the biological sample will cause the nanoneedles 310 to bend to different degrees. The maximum bending displacement (i.e., amplitude) of the tip of the nanoneedles 310 can be used to quantitatively and accurately assess the activity data of the biological sample.

[0048] The signal integration and processing module 400 is used to receive and process signals from the X-ray fluorescence imaging module 200 and the mechanospectral adhesion monitoring module 300, and to realize fluorescence visualization of the morphology and structural components of biological samples and the acquisition of bioactivity data. Specifically, the color image signal produced by the X-ray fluorescence imaging module 200 is analyzed and processed by the signal integration and processing module 400 to obtain the cell morphology, structure and composition information of the organism to be tested. In addition, the maximum bending displacement of the nanoneedle 310 recorded by the mechanospectral adhesion monitoring module 300 is transmitted to the signal integration and processing module 400 for processing and analysis to obtain the specific value of the bioactivity of the biological sample.

[0049] In other words, the biological detection device provided in this embodiment integrates adhesive mechanical spectrometry and X-ray excitation fluorescence imaging technology on the basis of contrast agents. It fuses the bioactivity and composition information obtained by adhesive mechanical spectrometry with the structural and composition information obtained by X-ray excitation fluorescence imaging, providing equipment support for the research and application of cross-scale, multi-mode, and visualized biological detection contrast agents that combine adhesive mechanical spectrometry and X-ray excitation fluorescence imaging, and promoting their development, optimization, and translational application in biomedical fields such as inflammation and infection, malignant diseases, and tumor diagnosis.

[0050] Preferably, the fluorescence signal excitation unit includes an X-ray excitation tube 210 that provides an X-ray radiation source for the biological sample. The X-ray excitation tube 210 is positioned facing the biological sample on the stage 100. An X-ray collimator 211 is provided at the exit end of the X-ray excitation tube 210. The X-ray collimator 211 is used to concentrate the dispersed laser emitted from the X-ray excitation tube 210 to form a parallel X-ray beam. Thus, the X-rays emitted by the X-ray excitation tube 210 can be successfully irradiated onto the biological sample and excite the contrast agent to generate a fluorescence signal.

[0051] It is worth mentioning here that, in this embodiment, the contrast agent is preferably set as NaAF4:B (A = Y, Zr, Nb, Mo, Tc, Fe, Mn, Bi; B = Ce, Nd, Ho, Er, Tm, Yb). When this type of contrast agent is excited by X-rays, it can produce color fluorescence changes from blue to green, red and even near-infrared, so as to facilitate the subsequent generation of color image signals at the image processing unit.

[0052] Preferably, the fluorescence signal processing unit includes:

[0053] Optical objective 220 is positioned to align with a biological sample on stage 100. Optical objective 220 is used to receive the fluorescence signal generated when a biological sample containing a contrast agent is excited by X-rays.

[0054] The filter group 230 is detachably installed in the fluorescence signal transmission channel and is used to intercept and filter fluorescence signals of different wavelengths.

[0055] An imaging camera 240 is disposed at the end of the fluorescence signal transmission channel. The imaging camera 240 is used to receive the fluorescence signal filtered by the filter group 230 and produce a color image signal.

[0056] Preferably, the filter group 230 includes a monochrome filter group or a color filter array. Specifically, in this embodiment, based on the self-developed contrast agent material NaAF4:B (A = Y, Zr, Nb, Mo, Tc, Fe, Mn, Bi; B = Ce, Nd, Ho, Er, Tm, Yb) which has the characteristics of multi-wavelength and tunable emission under X-ray excitation, the filter group 230 is configured to provide the function of monochrome filtering and color filtering.

[0057] Specifically, in this embodiment, the monochromatic filter group is set as a filter group 230 that allows different single-wavelength light (blue, green, red, near-infrared) to pass through. When the monochromatic filter group is selected, it can achieve the purpose of generating single-channel fluorescence imaging of blue, red, green, and near-infrared after the biological sample containing contrast agent is excited by X-rays.

[0058] The color filter array consists of a filter array with one blue, one red and two green filter elements arranged in a row. When the color filter array is selected, it can achieve the purpose of generating three primary color channels for multicolor imaging of biological samples containing contrast agents after X-ray excitation.

[0059] Preferably, a short-pass dichroic mirror 250 is provided between the filter group 230 and the imaging camera 240. Visible light is reflected when it shines on the short-pass dichroic mirror 250, while near-infrared light is transmitted when it shines on the short-pass dichroic mirror 250. Therefore, the short-pass dichroic mirror 250 is provided to isolate visible light from infrared light.

[0060] Preferably, the imaging camera 240 includes a visible light imaging camera 241 and a near-infrared imaging camera 242. In this embodiment, the visible light imaging camera 241 is set as a 400-900nm CCD camera, and the near-infrared imaging camera 242 is set as a 900-1700nm InGaAs camera. Specifically, when visible light and near-infrared light simultaneously illuminate the short-pass dichroic mirror 250, the visible light is reflected by the short-pass dichroic mirror 250 and received by the visible light imaging camera 241, while the near-infrared light is transmitted through the short-pass dichroic mirror 250 and received by the near-infrared imaging camera 242 to complete image isolation. The image is then transmitted to the signal integration and processing module 400 for image optimization and analysis.

[0061] Preferably, the mechanical spectroscopic adhesion monitoring module 300 also includes a mechanical motion signal acquisition unit 320 for detecting and recording the bending amplitude of the nanoparticles 310. Specifically, when the biological sample containing the contrast agent is placed on the stage 100, the nanoparticles 310 are first moved longitudinally by the microcomputer control so that the nanoparticles 310 are immersed in the biological sample.

[0062] The position of the nanoneedle 310 when no biological sample adheres is recorded as the equilibrium point, which is recorded by the mechanical motion signal acquisition device 320 and transmitted to the signal integration and processing module 400.

[0063] After the nano-needle 310 is co-cultured with the biological sample for a period of time, it bends due to its adhesive activity. The position of the bend is detected and recorded by the mechanical motion signal acquisition device 320 and transmitted to the signal processing module.

[0064] Preferably, the signal integration and processing module 400 includes a digital image processing terminal 410 and a mechanical motion signal analysis terminal 420, wherein:

[0065] The digital image processing terminal 410 is used to receive and process color image signals from the imaging camera 240. The digital image processing terminal 410 has the functions of image enhancement, image restoration, image encoding and image recognition.

[0066] Specifically, the digital image processing terminal 410 first performs image enhancement and image restoration sequentially on the received color image signal to eliminate interference and blurring of image quality, weaken background fluorescence, highlight the morphological structure information of biological cells in the image, and obtain the restored image. Subsequently, the image encoding utilizes the statistical characteristics of the image signal and the multi-wavelength tunable fluorescence optical characteristics generated by the contrast agent under X-ray excitation to efficiently encode the restored image signal, thereby identifying the component information in the biological sample imaging image.

[0067] Mechanical motion signal analysis terminal 420, which is used to receive and process data collected by mechanical motion signal acquisition device 320;

[0068] Specifically, the mechanical motion signal analysis terminal 420 can analyze and process the speed of signal change, the range of signal amplitude, and the continuity and discreteness of the signal. By identifying the amplitude, variation law, and waveform distortion characteristics of the signal time-domain waveform, it can perform quantitative data evaluation and dynamic continuous real-time monitoring of the activity status of biological samples.

[0069] In addition, during this process, the simple harmonic amplitude generated by the vibration of the periodic nanoneedle 310 submerged in random background noise signal is stripped and detected by the application of autocorrelation function analysis, which further improves the accuracy of the data.

[0070] This invention also provides a cross-scale, multi-modal, and visualized biological detection method, comprising steps S11-S14 and S21-S23, wherein S11-S14 and S21-S23 can be performed simultaneously. The specific steps include:

[0071] S11, X-ray excitation tube 210 emits X-rays to irradiate the surface of the biological sample. At this time, the biological sample containing the contrast agent is excited and produces fluorescence changes.

[0072] S12, the optical objective lens 220 recovers the fluorescence signal generated after the biological sample containing the contrast agent is excited, and intercepts and filters the fluorescence signals of different wavelengths through the filter group 230. The filtered fluorescence signal is transmitted to the imaging camera 240 and generates a color image signal, which is then transmitted to the digital image processing terminal 410.

[0073] S13, the digital image processing terminal 410 sequentially performs image enhancement and image restoration on the received color image signal, thereby eliminating interference and blurring of image quality, weakening background fluorescence, highlighting the morphological structure information of biological cells in the image, and obtaining the restored image.

[0074] S14, the digital image processing terminal 410 sequentially performs image encoding and image recognition on the restored image to identify the component information in the biological sample imaging image;

[0075] S21. The nano-needle 310 is immersed in the biological sample by an external microcomputer control, and the position of the nano-needle 310 when no biological sample adheres is recorded as the equilibrium point. The position of the equilibrium point is recorded by the mechanical motion signal acquisition device 320 and transmitted to the mechanical motion signal analysis terminal 420.

[0076] S22. After the nano-needle 310 has been cultured in the biological sample for a period of time, the nano-needle 310 bends due to the adhesive behavior of the biological sample. The position where the tip of the nano-needle 310 bends is recorded again by the mechanical motion signal acquisition device 320 and transmitted to the mechanical motion signal analysis terminal 420.

[0077] S23, the mechanical motion signal acquisition device 320 continuously acquires the bending amplitude of the tip of the nano-suspended needle 310 and transmits the signal to the mechanical motion signal analysis terminal 420. The mechanical motion signal analysis terminal 420 identifies the amplitude, variation law and waveform distortion signal characteristics of the signal time domain waveform, performs quantitative data evaluation and dynamic continuous real-time monitoring of the activity status of the biological sample, and obtains accurate biological sample activity data.

[0078] The technical means disclosed in this invention are not limited to those described above, but also include technical solutions composed of any combination of the above technical features. The above are specific embodiments of this invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of this invention, and these improvements and modifications are also considered within the scope of protection of this invention.

[0079] It should be noted that all directional indications (such as up, down, left, right, front, back, etc.) in the embodiments of the present invention are only used to explain the relative positional relationship and movement of each component in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indication will also change accordingly.

[0080] Furthermore, in this invention, descriptions involving terms such as "first," "second," and "a" are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0081] In this invention, unless otherwise explicitly specified and limited, the terms "connection," "fixed," etc., should be interpreted broadly. For example, "fixed" can mean a fixed connection, a detachable connection, or an integral part; it can mean a mechanical connection or an electrical connection; it can mean a direct connection or an indirect connection through an intermediate medium; it can mean the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0082] Furthermore, the technical solutions of the various embodiments of the present invention can be combined with each other, but only if they are feasible for those skilled in the art. If the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

Claims

1. A multi-scale, multi-modal, visualized biological detection device, characterized in that, include: Stage, used to hold biological samples containing contrast agents; The X-ray fluorescence imaging module is located on top of the stage. The X-ray fluorescence imaging module includes a fluorescence signal excitation unit, a fluorescence signal processing unit, and an image processing unit. The fluorescence signal excitation unit is used to emit X-rays that can irradiate biological samples. The fluorescence signal processing unit is used to collect and screen the fluorescence signals generated after the X-ray excitation of the contrast agent. The image processing unit is used to receive the screened fluorescence signals and generate color image signals. A mechanical spectroscopic adhesion monitoring module includes nanoneedles that can adhere to biological samples, and monitors the active adhesion of biological samples in real time by the degree of bending of the nanoneedles. The signal integration and processing module is used to receive, process and analyze signals from the X-ray fluorescence imaging module and the mechanospectral adhesion monitoring module, and to realize fluorescence visualization of biological sample morphology and structural components and acquisition of bioactivity data. The fluorescence signal processing unit includes: An optical objective lens is positioned to be aligned with the biological sample on the stage, and the optical objective lens is used to receive fluorescence signals; A filter array, which is detachably installed in the fluorescence signal transmission channel, is used to intercept and filter fluorescence signals of different wavelengths; The image processing unit includes an imaging camera, which is disposed at the end of the fluorescence signal transmission channel. The imaging camera is used to receive the fluorescence signal filtered by the filter group and produce a color image signal. A short-pass dichroic mirror is provided between the filter group and the imaging camera; The mechanospectral adhesion monitoring module also includes a mechanical motion signal acquisition device for detecting and recording the bending amplitude of the nanoneedle; The signal integration and processing module includes: A digital image processing terminal, used to receive and process the color image signals from the imaging camera; A mechanical motion signal analysis terminal is used to receive and process the data collected by the mechanical motion signal acquisition device.

2. The multi-scale, multi-modal, and visualized biological detection device according to claim 1, characterized in that, The fluorescence signal excitation unit includes an X-ray excitation tube that provides an X-ray radiation source for the biological sample. The X-ray excitation tube is positioned facing the biological sample on the stage, and an X-ray collimator is provided at the exit end of the X-ray excitation tube.

3. The multi-scale, multi-modal, and visualized biological detection device according to claim 1, characterized in that, The filter group includes a monochrome filter group or a color filter array.

4. The multi-scale, multi-modal, visualized biological detection device according to claim 1, characterized in that, The imaging camera includes a visible light imaging camera and a near-infrared imaging camera. The near-infrared light emitted by the biological sample after X-ray radiation is transmitted through the short-pass dichroic mirror and collected by the near-infrared imaging camera. The visible light emitted by the biological sample after X-ray radiation is reflected through the short-pass dichroic mirror and collected by the visible light imaging camera.

5. A cross-scale, multi-modal, visualized biological detection method, applied to the cross-scale, multi-modal, visualized biological detection device described in claim 2, comprising steps S11-S14 and S21-S23, wherein S11-S14 and S21-S23 can be performed simultaneously, characterized in that, The specific steps include: S11. The X-ray excitation tube emits X-rays to irradiate the surface of the biological sample. At this time, the biological sample containing the contrast agent is excited and produces a fluorescence change. S12. The optical objective lens recovers the fluorescence signal generated after the biological sample containing the contrast agent is excited, and intercepts and filters the fluorescence signals of different wavelengths through the filter group. The filtered fluorescence signal is transmitted to the imaging camera and generates a color image signal, which is then transmitted to the digital image processing terminal. S13. The digital image processing terminal sequentially performs image enhancement and image restoration on the received color image signal to eliminate interference and blurring of image quality, weaken background fluorescence, highlight the morphological structure information of biological cells in the image, and obtain the restored image. S14. The digital image processing terminal sequentially performs image encoding and image recognition on the restored image to identify the component information in the biological sample imaging image. S21. The nano-needle is immersed in the biological sample by an external microcomputer, and the position of the nano-needle when no biological sample adheres is recorded as the equilibrium point. The position of the equilibrium point is recorded by the mechanical motion signal acquisition device and transmitted to the mechanical motion signal analysis terminal. S22. After the nano-needle is cultured in the biological sample for a period of time, the nano-needle bends due to the adhesive behavior of the biological sample. The location where the tip of the nano-needle bends is recorded again by the mechanical motion signal acquisition device and transmitted to the mechanical motion signal analysis terminal. S23. The mechanical motion signal acquisition device continuously acquires the bending amplitude of the tip of the nano-needle and transmits the signal to the mechanical motion signal analysis terminal. The mechanical motion signal analysis terminal identifies the amplitude, variation law and waveform distortion characteristics of the signal time-domain waveform to perform quantitative data evaluation and dynamic continuous real-time monitoring of the activity status of the biological sample, and obtains accurate biological sample activity data.

Images