Novel antibacterial drug screening system based on near-infrared two-zone living body imaging

Abstract

The invention discloses a novel antibacterial drug screening system based on near-infrared two-zone living body imaging, and relates to the technical field of biomedicine. The system specifically comprises a control end hardware equipment layer which takes an STM32H743 single-chip microcomputer as a core controller and has the functions of controlling the height of a lifting platform, controlling the incident angle of an excitation light source, controlling the intensity of the light source, controlling a light source following system, designing a display screen man-machine interaction interface and developing functions, controlling a photoelectric / metal sensor, and controlling the control end hardware equipment layer; and the control end completes a bidirectional communication function with an upper computer through a serial interface USB. By utilizing the optical fiber coupled laser and SCMOS camera technology, the accuracy and efficiency of antibacterial drug screening are remarkably improved, real-time monitoring and accurate evaluation of the acting effect of the drug in a living body are realized, novel drugs with efficient antibacterial activity can be rapidly screened out, and the method is suitable for large-scale popularization and application. Powerful support is provided for treatment and research of infectious diseases.

Description

Technical Field

[0001] The invention relates to the field of biomedicine, and in particular to a novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging. Background Art

[0002] In the biomedical field, antimicrobial drug screening is a key step in the development of new antibiotics and the treatment of infectious diseases. Traditional antimicrobial drug screening methods often rely on in vitro experiments, which can preliminarily evaluate the antimicrobial activity of drugs, but it is difficult to accurately reflect the actual effect of drugs in vivo.

[0003] Existing antimicrobial drug screening methods mainly rely on in vitro experiments. Although this method can preliminarily evaluate the antimicrobial activity of drugs, it cannot accurately reflect the actual effect of drugs in vivo, especially key information such as the metabolism and distribution of drugs in complex biological environments and their interaction with target pathogens. In addition, traditional imaging technology has problems such as low resolution, limited penetration depth, and high background noise in in vivo imaging, making it difficult to achieve real-time monitoring and accurate evaluation of drug effects.

[0004] To this end, the present invention proposes a novel antimicrobial drug screening system based on near-infrared second-zone in vivo imaging. Summary of the invention

[0005] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a new antibacterial drug screening system based on near-infrared second-zone in vivo imaging.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A new antimicrobial drug screening system based on near-infrared zone II in vivo imaging, including:

[0008] The control end hardware device layer uses the STM32H743 single-chip microcomputer as the core controller, which has the functions of controlling the height of the lifting platform, controlling the incident angle of the excitation light source, controlling the light source intensity, controlling the light source following system, designing and developing the human-computer interaction interface of the display screen, controlling the photoelectric / metal sensor, and completing the two-way communication between the control end and the host computer through the serial interface USB;

[0009] Image acquisition layer, which consists of SCMOS industrial camera, optical path system, halogen light source, excitation light source, lens, filter and imaging system;

[0010] The transport layer is responsible for the transmission of data and control instructions when the lower computer and the upper computer communicate with each other, the transmission of control instructions and image data between the industrial camera and the upper computer, and the interactive transmission of various control instructions and data between the underlying development board and the hardware equipment;

[0011] The user application layer has the functions of displaying, storing, analyzing and processing image data and viewing historical data.

[0012] As a preferred embodiment of the present invention: the control end hardware device layer includes a voltage-stabilized power supply module, an MCU main controller, a sensor module, a storage module, a time-gated imaging module, a lifting platform module, a light source following module, a halogen lamp control module and a human-computer interaction module.

[0013] As a preferred embodiment of the present invention: the voltage-stabilized power supply module is used to provide a stable voltage to the MCU main controller, the sensor module, the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module; the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module are connected to the MCU main control via serial port communication; and the time-gated imaging module and the sensor module are command-controlled via the MCU main controller.

[0014] As a preferred embodiment of the present invention: the excitation light source of the image acquisition layer is a fiber-coupled laser, and the output wavelengths thereof are 808 nm and 980 nm respectively.

[0015] As a preferred embodiment of the present invention, the imaging system includes an image acquisition unit, a timing control unit, a data cache unit, a data transmission unit, a host computer control unit and an image processing unit.

[0016] As a preferred embodiment of the present invention: the image acquisition unit is a LUX13HS image sensor;

[0017] The timing control unit is Kintex-7;

[0018] The data cache unit is DDR3 SDRAM;

[0019] The data transmission unit is a PCIE interface.

[0020] As a preferred embodiment of the present invention: the working logic of the image acquisition layer includes the following steps:

[0021] A1: The laser emits laser light, which is expanded by a beam expander to form uniform parallel light;

[0022] A2: The incident angle of the laser and the position of the experimental animal stage are adjusted through the imaging system software on the upper computer or the human-computer interactive display screen on the lower computer, so that the laser can be evenly irradiated on the small animal;

[0023] A3: The laser excites the fluorescent probe injected into the small animal, emitting fluorescent photons. The fluorescent photons are absorbed and reflected by the biological tissue and then escape from the surface. The industrial camera is connected and opened through the software system on the host computer.

[0024] A4: Adjust the exposure time, gain and camera focus of the industrial camera, and display and save the image data signal after obtaining it.

[0025] As a preferred embodiment of the present invention: the working logic of the imaging system includes the following steps:

[0026] B1: The acquisition unit collects the incident light signal and converts it into a digital signal that can be used by subsequent circuits;

[0027] B2: The timing control unit is responsible for providing timing for the image acquisition unit, data cache unit, and data transmission unit, and coordinating the normal operation of the entire system;

[0028] B3: The data cache module is responsible for temporarily storing image data;

[0029] B4: The data transmission unit transmits image data to the PC and receives control parameters sent by the PC;

[0030] B5: The host computer control unit is responsible for sensor control parameter setting, data transmission interface detection and data collection, and the entire system activity status display;

[0031] B6: The image processing unit processes the collected images.

[0032] As a preferred embodiment of the present invention: after the image acquisition layer acquires the image, the method for image processing comprises the following steps:

[0033] C1: Image segmentation, which divides the input image into equal-sized and non-overlapping sub-blocks;

[0034] C2: Preprocess the sub-blocks and calculate their histogram h(x), clipping threshold, and mapping relationship. The preprocessing formula is: Among them, CL is the clipping threshold, CL k is the shear coefficient, M x and M y are the number of pixels in the x and y directions of each small block after segmentation, and N is the number of gray levels;

[0035] C3: Redistribute the pixels, cut the histogram grayscale that exceeds the clipping threshold CL, and evenly distribute the excess part to each grayscale level;

[0036] C4: The grayscale values ​​of pixels are reconstructed to obtain the final image.

[0037] As a preferred embodiment of the present invention: the C3 step comprises the following steps:

[0038] C31: Assign S pixels exceeding the clipping threshold to each sub-block, the number of pixels is S mean ;

[0039] C32: The histogram after histogram equalization is H(x), and its formula is:

[0040]

[0041] The beneficial effects of the present invention are:

[0042] 1. The present invention significantly improves the accuracy and efficiency of antimicrobial drug screening by utilizing fiber-coupled laser and SCMOS camera technology, realizes real-time monitoring and accurate evaluation of drug effects in vivo, helps to quickly screen out new drugs with high antimicrobial activity, and provides strong support for the treatment and research of infectious diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 This is a schematic diagram of the hardware system structure of a novel antimicrobial drug screening system based on near-infrared second-zone in vivo imaging proposed by the present invention;

[0044] Figure 2 This is a schematic diagram of the imaging system architecture of a novel antimicrobial drug screening system based on near-infrared second-zone in vivo imaging proposed by the present invention;

[0045] Figure 3 This is a working state transition diagram of an imaging system of a novel antimicrobial drug screening system based on near-infrared second-zone in vivo imaging proposed by the present invention;

[0046] Figure 4 This is the overall development flow chart of the host computer system software of a new antimicrobial drug screening system based on near-infrared second-zone in vivo imaging proposed by the present invention;

[0047] Figure 5 This is a functional diagram of the interface system of the host computer of a novel antimicrobial drug screening system based on near-infrared two-zone in vivo imaging proposed by the present invention;

[0048] Figure 6 This is a main program development flow chart for the system host computer side of a new antimicrobial drug screening system based on near-infrared second-zone in vivo imaging proposed by the present invention. DETAILED DESCRIPTION

[0049] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0050] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0051] Embodiment 1:

[0052] A novel antimicrobial drug screening system based on near-infrared zone II in vivo imaging, comprising:

[0053] The control end hardware device layer uses the STM32H743 single-chip microcomputer as the core controller, which has the functions of controlling the height of the lifting platform, controlling the incident angle of the excitation light source, controlling the light source intensity, controlling the light source following system, designing and developing the human-computer interaction interface of the display screen, controlling the photoelectric / metal sensor, and completing the two-way communication between the control end and the host computer through the serial interface USB;

[0054] Image acquisition layer, which consists of SCMOS industrial camera, optical path system, halogen light source, excitation light source, lens, filter and imaging system;

[0055] The transport layer is responsible for the transmission of data and control instructions when the lower computer and the upper computer communicate with each other, the transmission of control instructions and image data between the industrial camera and the upper computer, and the interactive transmission of various control instructions and data between the underlying development board and the hardware equipment;

[0056] The user application layer has the functions of displaying, storing, analyzing and processing image data and viewing historical data.

[0057] The control end hardware device layer includes a voltage-stabilized power supply module, an MCU main controller, a sensor module, a storage module, a time-gated imaging module, a lifting platform module, a light source following module, a halogen lamp control module and a human-computer interaction module.

[0058] The voltage-stabilized power supply module is used to provide a stable voltage to the MCU main controller, the sensor module, the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module. The lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module are connected to the MCU main control via serial port communication, and the time-gated imaging module and the sensor module are command-controlled via the MCU main controller.

[0059] The excitation light source of the image acquisition layer is a fiber-coupled laser, and its output wavelengths are 808nm and 980nm respectively. The wavelength power output performance is stable and continuously adjustable. The auxiliary optical fiber equipped with it can install the optical fiber head on the rotating device of the servo, and adjust the incident angle of the excitation light source by controlling the rotation direction of the servo. In this way, the light source of the laser emission can also be enclosed in the dark box of the living body imaging, which can ensure that the entire experiment is performed in the dark box, avoiding interference from unnecessary external light, reducing environmental background noise, and enabling the excited fluorescence signal to be effectively detected.

[0060] The imaging system comprises an image acquisition unit, a timing control unit, a data cache unit, a data transmission unit, a host computer control unit and an image processing unit.

[0061] Embodiment 2:

[0062] A novel antimicrobial drug screening system based on near-infrared zone II in vivo imaging, comprising:

[0063] The control end hardware device layer uses the STM32H743 single-chip microcomputer as the core controller, which has the functions of controlling the height of the lifting platform, controlling the incident angle of the excitation light source, controlling the light source intensity, controlling the light source following system, designing and developing the human-computer interaction interface of the display screen, controlling the photoelectric / metal sensor, and completing the two-way communication between the control end and the host computer through the serial interface USB;

[0064] Image acquisition layer, which consists of SCMOS industrial camera, optical path system, halogen light source, excitation light source, lens, filter and imaging system;

[0065] The transport layer is responsible for the transmission of data and control instructions when the lower computer and the upper computer communicate with each other, the transmission of control instructions and image data between the industrial camera and the upper computer, and the interactive transmission of various control instructions and data between the underlying development board and the hardware equipment;

[0066] The user application layer has the functions of displaying, storing, analyzing and processing image data and viewing historical data.

[0067] The control end hardware device layer includes a voltage-stabilized power supply module, an MCU main controller, a sensor module, a storage module, a time-gated imaging module, a lifting platform module, a light source following module, a halogen lamp control module and a human-computer interaction module.

[0068] The voltage-stabilized power supply module is used to provide a stable voltage to the MCU main controller, the sensor module, the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module. The lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module are connected to the MCU main control via serial port communication, and the time-gated imaging module and the sensor module are command-controlled via the MCU main controller.

[0069] The excitation light source of the image acquisition layer is a fiber-coupled laser, and its output wavelengths are 808nm and 980nm respectively. The wavelength power output performance is stable and continuously adjustable. The auxiliary optical fiber equipped with it can install the optical fiber head on the rotating device of the servo, and adjust the incident angle of the excitation light source by controlling the rotation direction of the servo. In this way, the light source of the laser emission can also be enclosed in the dark box of the living body imaging, which can ensure that the entire experiment is performed in the dark box, avoiding interference from unnecessary external light, reducing environmental background noise, and enabling the excited fluorescence signal to be effectively detected.

[0070] The imaging system comprises an image acquisition unit, a timing control unit, a data cache unit, a data transmission unit, a host computer control unit and an image processing unit.

[0071] The image acquisition unit is a LUX13HS image sensor, which has a 1M pixel, 4000+fps, global shutter CMOS digital sensor. The chip features low-noise pixels and can obtain higher frame rates by opening windows. It also supports RGB color or monochrome.

[0072] The LUX13HS sensor can support a master clock input of 100MHz to 166MHz. Under a master clock of 166MHz, it can reach a frame rate of 4000fps at a full-frame effective pixel of 1280*864. The design of the present invention is for SCMOS sensors, and a master clock input of 100MHz is selected. Under a master clock of 100M, the sensor can reach a maximum frame rate of 2800fps, which meets the purpose of high-speed imaging;

[0073] The timing control unit is Kintex-7, which can provide high-density logic, high-performance transceivers, memory and other functions, has a high cost performance, and provides a balanced and optimized configuration for high-end functions;

[0074] The data cache unit is DDR3 SDRAM, which has the advantages of high speed, large capacity, stable operation and high cost performance. The selection of DDR3 is mainly based on two points: internal capacity and operating frequency. Since LUX13HS has 80 pairs of LVDS data output ports, each LVDS port in this design transmits 4 bits in serial within one clock at a clock frequency of 100MHz, that is, the LVDS data rate is 400MHz, and the chip has two channels that output data almost simultaneously. After the data output by these two channels is converted from serial to parallel and regularized, each channel needs to transmit 160bit data at 100MHz, and the maximum bandwidth is 160bit*2*100M / s=31.25Gb / s. In order to ensure that such a high-speed data stream can be fully received and stored, the frequency and bandwidth parameters must be paid attention to when selecting a DDR3 chip. This design uses Micron's MT41J64M16, which uses 4 DDR3 particles to form a 64-bit 1GB cache, and its maximum operating clock frequency can reach 667MHz. This system uses 500MHz as the working clock of DDR3, and its theoretical transmission bandwidth can reach 64bit*2*500M / s=62.5Gb / s (DDR3 upper and lower edges work simultaneously). This design uses the DDR3 controller IP core provided by Xilinx to perform DDR3 read and write operations. Generally, when performing continuous write operations on DDR3, its bandwidth utilization can reach about 60%, that is, the DDR3 write bandwidth can reach 37.5Gb / s. It can be seen that the bandwidth meets the requirements of this SCMOS high-speed imaging system;

[0075] The data transmission unit is a PCIE interface. PCIE is a third-generation I / O interconnect bus technology. It supports point-to-point serial connections between chips and devices, and greatly improves the effective data bandwidth of the bus with low overhead, low latency and other characteristics. It supports power management, quality of service, hot plugging of devices, and error handling. At present, the PCI-SIG organization has released multiple PCIE standards. According to the unidirectional link bandwidth, PCIE1.x (PCIEGen1) uses a rate of 2.5Gbps, PCIE2.x (PCIEGen2) uses 5.0Gbps, and the latest PCIE3.0 supports data transmission at a rate of up to 8.0Gbps. In terms of transmission rate, the PCIE bus interface is the fastest. If PCIE2.0x8 is used, the speed can reach 40Gbps in theory. And because the FPGAKintex-7 used in the present invention directly supports the PCIEEndpoint hard core, the system finally selects the PCIE interface as the interface for data transmission with the PC.

[0076] Embodiment 3:

[0077] A novel antimicrobial drug screening system based on near-infrared zone II in vivo imaging, comprising:

[0078] The control end hardware device layer uses the STM32H743 single-chip microcomputer as the core controller, which has the functions of controlling the height of the lifting platform, controlling the incident angle of the excitation light source, controlling the light source intensity, controlling the light source following system, designing and developing the human-computer interaction interface of the display screen, controlling the photoelectric / metal sensor, and completing the two-way communication between the control end and the host computer through the serial interface USB;

[0079] Image acquisition layer, which consists of SCMOS industrial camera, optical path system, halogen light source, excitation light source, lens, filter and imaging system;

[0080] The transport layer is responsible for the transmission of data and control instructions when the lower computer and the upper computer communicate with each other, the transmission of control instructions and image data between the industrial camera and the upper computer, and the interactive transmission of various control instructions and data between the underlying development board and the hardware equipment;

[0081] The user application layer has the functions of displaying, storing, analyzing and processing image data and viewing historical data.

[0082] The control end hardware device layer includes a voltage-stabilized power supply module, an MCU main controller, a sensor module, a storage module, a time-gated imaging module, a lifting platform module, a light source following module, a halogen lamp control module and a human-computer interaction module.

[0083] The voltage-stabilized power supply module is used to provide a stable voltage to the MCU main controller, the sensor module, the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module. The lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module are connected to the MCU main control via serial port communication, and the time-gated imaging module and the sensor module are command-controlled via the MCU main controller.

[0084] The excitation light source of the image acquisition layer is a fiber-coupled laser, and its output wavelengths are 808nm and 980nm respectively. The wavelength power output performance is stable and continuously adjustable. The auxiliary optical fiber equipped with it can install the optical fiber head on the rotating device of the servo, and adjust the incident angle of the excitation light source by controlling the rotation direction of the servo. In this way, the light source of the laser emission can also be enclosed in the dark box of the living body imaging, which can ensure that the entire experiment is performed in the dark box, avoiding interference from unnecessary external light, reducing environmental background noise, and enabling the excited fluorescence signal to be effectively detected.

[0085] The imaging system comprises an image acquisition unit, a timing control unit, a data cache unit, a data transmission unit, a host computer control unit and an image processing unit.

[0086] The image acquisition unit is a LUX13HS image sensor, which has a 1M pixel, 4000+fps, global shutter CMOS digital sensor. The chip features low-noise pixels and can obtain higher frame rates by opening windows. It also supports RGB color or monochrome.

[0087] The LUX13HS sensor can support a master clock input of 100MHz to 166MHz. Under a master clock of 166MHz, it can reach a frame rate of 4000fps at a full-frame effective pixel of 1280*864. The design of the present invention is for SCMOS sensors, and a master clock input of 100MHz is selected. Under a master clock of 100M, the sensor can reach a maximum frame rate of 2800fps, which meets the purpose of high-speed imaging;

[0088] The timing control unit is Kintex-7, which can provide high-density logic, high-performance transceivers, memory and other functions, has a high cost performance, and provides a balanced and optimized configuration for high-end functions;

[0089] The data cache unit is DDR3 SDRAM, which has the advantages of high speed, large capacity, stable operation and high cost performance. The selection of DDR3 is mainly based on two points: internal capacity and operating frequency. Since LUX13HS has 80 pairs of LVDS data output ports, each LVDS port in this design transmits 4 bits in serial within one clock at a clock frequency of 100MHz, that is, the LVDS data rate is 400MHz, and the chip has two channels that output data almost simultaneously. After the data output by these two channels is converted from serial to parallel and regularized, each channel needs to transmit 160bit data at 100MHz, and the maximum bandwidth is 160bit*2*100M / s=31.25Gb / s. In order to ensure that such a high-speed data stream can be fully received and stored, the frequency and bandwidth parameters must be paid attention to when selecting a DDR3 chip. This design uses Micron's MT41J64M16, which uses 4 DDR3 particles to form a 64-bit 1GB cache, and its maximum operating clock frequency can reach 667MHz. This system uses 500MHz as the working clock of DDR3, and its theoretical transmission bandwidth can reach 64bit*2*500M / s=62.5Gb / s (DDR3 upper and lower edges work simultaneously). This design uses the DDR3 controller IP core provided by Xilinx to perform DDR3 read and write operations. Generally, when performing continuous write operations on DDR3, its bandwidth utilization can reach about 60%, that is, the DDR3 write bandwidth can reach 37.5Gb / s. It can be seen that the bandwidth meets the requirements of this SCMOS high-speed imaging system;

[0090] The data transmission unit is a PCIE interface. PCIE is a third-generation I / O interconnect bus technology. It supports point-to-point serial connections between chips and devices, and greatly improves the effective data bandwidth of the bus with low overhead, low latency and other characteristics. It supports power management, quality of service, hot plugging of devices, and error handling. At present, the PCI-SIG organization has released multiple PCIE standards. According to the unidirectional link bandwidth, PCIE1.x (PCIEGen1) uses a rate of 2.5Gbps, PCIE2.x (PCIEGen2) uses 5.0Gbps, and the latest PCIE3.0 supports data transmission at a rate of up to 8.0Gbps. In terms of transmission rate, the PCIE bus interface is the fastest. If PCIE2.0x8 is used, the speed can reach 40Gbps in theory. And because the FPGAKintex-7 used in the present invention directly supports the PCIEEndpoint hard core, the system finally selects the PCIE interface as the interface for data transmission with the PC.

[0091] The working logic of the image acquisition includes the following steps:

[0092] A1: The laser emits laser light, which is expanded by a beam expander to form uniform parallel light;

[0093] A2: The incident angle of the laser and the position of the experimental animal stage are adjusted through the imaging system software on the upper computer or the human-computer interactive display screen on the lower computer, so that the laser can be evenly irradiated on the small animal;

[0094] A3: The laser excites the fluorescent probe injected into the small animal, emitting fluorescent photons. The fluorescent photons escape from the surface after being absorbed and reflected by the biological tissue, and are connected and opened by the industrial camera through the software system on the host computer.

[0095] A4: Adjust the exposure time, gain and camera focus of the industrial camera, and display and save the image data signal after obtaining it.

[0096] The advantage of this system optical path design is that the excitation light source can be evenly irradiated onto the experimental biological samples from multiple angles, and the incident angle of the excitation light source can also be simply and effectively controlled. However, when the excitation light source is irradiated onto the surface of biological tissues, such as animal skin and hair, it will also produce strong spontaneous fluorescence, which will also interfere with the fluorescence signal excited by the fluorescent probe. In the design of the present invention, the role of the excitation light source is to excite the fluorescent probe in the tissue. In order to collect fluorescence in living animals, the power of the excitation light source needs to reach a certain intensity.

[0097] The working logic of the imaging system includes the following steps:

[0098] B1: The acquisition unit collects the incident light signal and converts it into a digital signal that can be used by subsequent circuits;

[0099] B2: The timing control unit is responsible for providing timing for the image acquisition unit, data cache unit, and data transmission unit, and coordinating the normal operation of the entire system;

[0100] B3: The data cache module is responsible for temporarily storing image data, which directly relieves the pressure of the interface part;

[0101] B4: The data transmission unit transmits image data to the PC and receives control parameters sent by the PC;

[0102] B5: The host computer control unit is responsible for sensor control parameter setting, data transmission interface detection and data collection, and the entire system activity status display;

[0103] B6: The image processing unit processes the collected images to achieve the effect of image noise reduction and enhancement to facilitate subsequent observation.

[0104] When collecting images, the external environment or the image collector's own problems may affect the collection of fluorescent signals, so that the output image has noise and low image clarity. Therefore, it is necessary to denoise and enhance the output image to improve the use value of the image. In view of the situation that the image data collected in the near-infrared zone II may have bright areas where there are fluorescent dyes and dark areas in other parts, the use of traditional image processing methods such as ordinary histogram equalization may cause excessive contrast enhancement in areas with concentrated grayscale, thereby forming new background noise. Therefore, the contrast-limited adaptive histogram equalization algorithm (CLAHE) is used in this system to process the image. The CLAH algorithm is a very effective histogram equalization algorithm. Compared with ordinary histogram equalization (HE) and adaptive histogram equalization (AHE) algorithms, it can limit the regional contrast and use interpolation methods to speed up the calculation speed. It can effectively enhance and improve the contrast of the image while effectively suppressing the background noise. In addition, the algorithm has a very strong enhancement effect on biomedical infrared images; after the image acquisition layer collects the image, the method for image processing includes the following steps:

[0105] C1: Image segmentation, which divides the input image into equal-sized and non-overlapping sub-blocks;

[0106] C2: Preprocess the sub-blocks and calculate their histogram h(x), clipping threshold, and mapping relationship. The preprocessing formula is: Among them, CL is the clipping threshold, CL k is the shear coefficient, M x and M yare the number of pixels in the x and y directions of each small block after segmentation, and N is the number of gray levels;

[0107] C3: Redistribute the pixels, cut the histogram grayscale that exceeds the clipping threshold CL, and evenly distribute the excess part to each grayscale level;

[0108] C4: The final image is reconstructed by the grayscale values ​​of the pixels. When running the algorithm, a clearer image can be obtained by adjusting its parameters (block size, number of blocks, etc.). These can be selected and controlled by the buttons on the main interface of the system. A clear image can be obtained after the original image has been processed.

[0109] The C3 step comprises the following steps:

[0110] C31: Assign S pixels exceeding the clipping threshold to each sub-block, the number of pixels is S mean ;

[0111] C32: The histogram after histogram equalization is H(x), and its formula is:

[0112]

[0113] Overall design of the imaging system host computer software:

[0114] The host computer software platform is the control center of the entire system and the interface between the user and the system. Experimenters can use this platform to conduct imaging experiments and complete the collection and processing of image data. Considering the practicality of the software and the good interactive experience of users during development. The software platform designed by the present invention adopts the development idea of ​​separating UI interface design from business logic functions. The entire design and development of the host computer module includes the design of the UI interface and the development of the function code in the main program.

[0115] For the development of upper computer software, first configure the relevant development environment, mainly including the configuration of external tools QtDesigner, PyUic, and PyRcc; then design the UI interface in the external tool QtDesigner; secondly, layout the controls of the designed interface in QtDesigner and add signals to the controls, and convert the .ui file saved in the UI interface into a .py file through the external tool PyUic. The conversion of py files can also be done through the command line; finally, create a main file, import the UI design interface file in the main file, and write the main logic function code.

[0116] (1) Display function. The host computer interface display module is used to receive user instructions and perform corresponding operations to display the current working status of the system, including the display of camera status and the display of collected image data.

[0117] (2) Control function of external devices. This part establishes a communication mechanism between the host computer and the core controller STM32H743 microcontroller on the lower computer side, and can send and receive data or corresponding instructions with the lower computer, thereby realizing the control of various hardware devices of the system on the host PC side, mainly including displacement stage control, light source intensity control, excitation light source incident angle control, photoelectric sensors, metal sensors and other sensors

[0118] Control, light source following system regulation, etc.

[0119] (3) Industrial camera control function. In the camera control module, the computer and the industrial camera establish a communication mechanism through the corresponding communication protocol to complete the adjustment of camera parameters and status display, as well as the call of the camera and the capture, storage, processing and display of image data. In addition, it also includes the viewing and display of stored historical data.

[0120] (4) Communication function. The host computer, control terminal, and camera all use corresponding communication protocols to complete information transmission and interaction. The communication function needs to establish a stable communication connection between the host computer and the slave computer to ensure the correct reception and transmission of control commands. The main communication methods used here are RS-485 serial interface, USB serial interface, and GigE Gigabit network.

[0121] Upper computer UI interface design:

[0122] The present invention uses Python to develop the host computer software and design the interface. Programming can well realize the drive and control of hardware, the regulation of industrial cameras, and the processing and analysis of image data. The software design of the host computer needs to be cross-platform interactive.

[0123] The main program design consists of two parts, namely the design of the user registration and login interface and the design of the main interface for system debugging and display. First, design each interface separately; then, create a Python file to connect the interfaces through custom signals. The login interface is used to complete user registration and login. The main interface of the system needs to complete the communication between the upper computer and the lower computer microcontroller and the camera, and can realize the transmission and reception of data and command information between the microcontroller and the microcontroller, as well as the function embedding of the secondary development of the industrial camera, so as to achieve the good control of the camera using the interface, and display, process and store the collected image data.

[0124] The development and design of the main program mainly adopts the combination of signals and slot functions in PyQt5. Signals and slots are the core information transmission mechanism in Qt, and are also the communication mechanism between objects in PyQt5 programming, which can establish connections between objects without interfering with each other. The advantages of programming design using the signal and slot mechanism are:

[0125] (1) When a signal is called, Qt can find all the slot functions bound to this signal function, which is more efficient and flexible than using the traditional callback function mechanism.

[0126] (2) The signal and slot mechanism can weaken the coupling of Qt objects. When the program is running, even if an associated object is deleted, it will not cause the program to crash.

[0127] (3) Type safety: Signals and slots can only be connected successfully if the type and number of parameters match, which increases the safety of code programming.

[0128] In PyQt5, every widget that inherits Qwidget can use the signal and slot mechanism. When a signal is emitted, the connected slot function will be automatically executed. The connection method is to connect the signal of one object to the slot function of another object through QObject.signal.connect. After establishing the connection between objects, users only need to write program code in the corresponding slot function according to their needs.

[0129] The design method of the present invention can be used for screening the following antibacterial drugs:

[0130] 1. Rapidly detect the in vivo effects of other antimicrobial drugs and vaccines (such as MARSA, Escherichia coli, Pseudomonas aeruginosa, Streptococcus pneumoniae, etc.).

[0131] After the drug has been tested in vitro and its stability and chemical properties have been understood, it can be tested in animals to effectively observe the absorption, distribution and metabolism of the drug in the animal body. At the same time, the experimental body can be dissected in a timely and effective manner to observe the effects of the drug on different tissues and organs and the pathways (targets) of the drug. It can effectively evaluate the effectiveness, safety and other indicators of the drug. It can help determine

[0132] Safe dosage range for drug application to avoid damage caused by overdose.

[0133] 2. Rapidly detect the in vivo effects of antiviral (such as influenza virus) drugs, vaccines, and antibodies.

[0134] Because this technology has higher sensitivity, it can detect drug efficacy more accurately than traditional methods. This technology can detect drug efficacy earlier and more sensitively. By giving inoculated mice different doses, different administration times, and different administration routes, the best administration route, dosage, and time of antiviral drugs can be observed, so as to formulate appropriate dosage forms and medication times.

[0135] 3. Efficient study of virulence-related genes of Mycobacterium tuberculosis (Mtb).

[0136] By marking genes related to drug metabolism and studying the effects of different drugs on the expression of the gene, we can indirectly know the metabolism of related drugs in the body. In pharmaceutical research, the luciferase reporter gene plasmid can be directly loaded into the vector to observe the targeted organs and distribution patterns of the drug vector in the body. In pharmacology, the target gene is marked with the luciferase gene to observe the pathway of drug action.

[0137] 4. Rapidly detect the in vivo effects of anti-tumor drugs.

[0138] This technology can image and track the biological behavior of living tissues or small animals under nearly non-invasive conditions, and has been widely used in tumor research. This technology enables researchers to directly and quickly measure tumor growth, metastasis, and drug response in various cancer models. Its characteristic is that its extremely high sensitivity allows even tiny tumor lesions (as few as a few hundred cells) to be detected. The detection sensitivity is greatly improved compared to traditional methods, and it is very suitable for quantitative analysis of tumor growth in vivo. It avoids inter-group differences caused by slaughtering mice and saves animal costs.

[0139] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A novel antibacterial drug screening system based on near-infrared zone II in vivo imaging, characterized in that: include: The control end hardware device layer uses the STM32H743 single-chip microcomputer as the core controller, which has the functions of controlling the height of the lifting platform, controlling the incident angle of the excitation light source, controlling the light source intensity, controlling the light source following system, designing and developing the human-computer interaction interface of the display screen, controlling the photoelectric / metal sensor, and completing the two-way communication between the control end and the host computer through the serial interface USB; Image acquisition layer, which consists of SCMOS industrial camera, optical path system, halogen light source, excitation light source, lens, filter and imaging system; The transport layer is responsible for the transmission of data and control instructions when the lower computer and the upper computer communicate with each other, the transmission of control instructions and image data between the industrial camera and the upper computer, and the interactive transmission of various control instructions and data between the underlying development board and the hardware equipment; The user application layer has the functions of displaying, storing, analyzing and processing image data and viewing historical data.

2. According to claim 1, a novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging is characterized in that: The control end hardware device layer includes a voltage-stabilized power supply module, an MCU main controller, a sensor module, a storage module, a time-gated imaging module, a lifting platform module, a light source following module, a halogen lamp control module and a human-computer interaction module.

3. A novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 2, characterized in that: The voltage-stabilized power supply module is used to provide a stable voltage to the MCU main controller, the sensor module, the lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module. The lifting platform module, the light source following module, the halogen lamp control module and the human-computer interaction module are connected to the MCU main control via serial port communication, and the time-gated imaging module and the sensor module are command-controlled via the MCU main controller.

4. A novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 1, characterized in that: The excitation light source of the image acquisition layer is a fiber-coupled laser, and the output wavelengths thereof are 808 nm and 980 nm respectively.

5. According to claim 1, a novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging is characterized in that: The imaging system comprises an image acquisition unit, a timing control unit, a data cache unit, a data transmission unit, a host computer control unit and an image processing unit.

6. A novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 5, characterized in that: The image acquisition unit is a LUX13HS image sensor; The timing control unit is Kintex-7; The data cache unit is DDR3 SDRAM; The data transmission unit is a PCIE interface.

7. The novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 1 is characterized in that: The working logic of the image acquisition layer includes the following steps: A1: The laser emits laser light, which is expanded by a beam expander to form uniform parallel light; A2: The incident angle of the laser and the position of the experimental animal stage are adjusted through the imaging system software on the upper computer or the human-computer interactive display screen on the lower computer, so that the laser can be evenly irradiated on the small animal; A3: The laser excites the fluorescent probe injected into the small animal, emitting fluorescent photons. The fluorescent photons are absorbed and reflected by the biological tissue and then escape from the surface. The industrial camera is connected and opened through the software system on the host computer. A4: Adjust the exposure time, gain and camera focus of the industrial camera, and display and save the image data signal after obtaining it.

8. The novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 1 is characterized in that: The working logic of the imaging system includes the following steps: B1: The acquisition unit collects the incident light signal and converts it into a digital signal that can be used by subsequent circuits; B2: The timing control unit is responsible for providing timing for the image acquisition unit, data cache unit, and data transmission unit, and coordinating the normal operation of the entire system; B3: The data cache module is responsible for temporarily storing image data; B4: The data transmission unit transmits image data to the PC and receives control parameters sent by the PC; B5: The host computer control unit is responsible for sensor control parameter setting, data transmission interface detection and data collection, and the entire system activity status display; B6: The image processing unit processes the collected images.

9. The novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 1 is characterized in that: After the image acquisition layer acquires the image, the method for image processing comprises the following steps: C1: Image segmentation, which divides the input image into equal-sized and non-overlapping sub-blocks; C2: Preprocess the sub-blocks and calculate their histogram h(x), clipping threshold, and mapping relationship. The preprocessing formula is: Among them, CL is the clipping threshold, CL k is the shear coefficient, M x and M y are the number of pixels in the x and y directions of each small block after segmentation, and N is the number of gray levels; C3: Redistribute the pixels, cut the histogram grayscale that exceeds the clipping threshold CL, and evenly distribute the excess part to each grayscale level; C4: The grayscale values ​​of pixels are reconstructed to obtain the final image.

10. A novel antibacterial drug screening system based on near-infrared second-zone in vivo imaging according to claim 9, characterized in that: The C3 step comprises the following steps: C31: Assign S pixels exceeding the clipping threshold to each sub-block, the number of pixels is S mean ; C32: The histogram after histogram equalization is H(x), and its formula is:

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