Rapid high-resolution biological imprinting optical imaging system

Abstract

The invention discloses a rapid high-resolution biological imprinting optical imaging system, and belongs to the technical field of biomedical detection and molecular imaging. The system comprises a main control computer, a high-precision motion platform, a multi-mode imaging cabin and an embedded control core. The imaging cabin integrates a deep refrigeration sCMOS detector, an intelligent temperature control tabletting module, a multispectral LED light source and an automatic filter wheel. High-sensitivity and low-noise imaging is realized by controlling the working temperature of the sCMOS sensor to be below-45 DEG C and combining accurate regulation and control of the sample temperature. The system supports rapid switching of multiple imaging modes such as chemiluminescence and multicolor fluorescence, and is equipped with automatic focusing and intelligent monitoring functions. The defects of an existing imaging system in the aspects of detection speed, resolution and multi-modal detection are overcome, and a high-performance solution is provided for biological detection of western blot, nucleic acid blot and the like.

Description

Technical Field

[0001] This invention relates to the field of biomedical detection and molecular imaging technology, specifically a rapid high-resolution bioimprint optical imaging system. Background Technology

[0002] Bioblotting is an indispensable analytical technique in molecular biology, clinical diagnostics, and drug development. Its core principle is to separate and transfer biological macromolecules (such as proteins, DNA, and RNA) onto a solid membrane via electrophoresis, label them using specific probes or antibodies, and finally detect the signal using an imaging system. The performance of the imaging system directly determines the sensitivity, resolution, and quantitative accuracy of the detection.

[0003] Currently, mainstream imprint imaging systems mainly use the following types of detectors: CCD detectors, while possessing high quantum efficiency, suffer from drawbacks such as high dark current noise, slow readout speed, large size, and high cost. Especially under long-term exposure conditions, dark current accumulation leads to a significant decrease in signal-to-noise ratio.

[0004] EMCCD detectors: improve low-light detection capabilities through electron multiplication, but are extremely expensive and also suffer from slow reading speed and size issues.

[0005] CMOS detectors have seen rapid development in recent years, offering advantages such as fast readout speed, high integration, low cost, and low power consumption. However, ordinary CMOS detectors still exhibit significant dark current noise at room temperature, limiting their application in extremely low light detection.

[0006] PMT detectors are mainly used for scanning imaging of chemiluminescence. Although they have high sensitivity, they are point-by-point scanning devices with slow imaging speed, making them unsuitable for fast, high-throughput applications.

[0007] In addition, existing imprint imaging systems have common shortcomings in the following aspects: The trade-off between resolution and sensitivity: High resolution often requires small pixels, but small pixels reduce the light-receiving area per pixel, thus decreasing sensitivity. Improving low-light signal capture capability while maintaining high resolution is a major challenge.

[0008] Imaging speed: For signals such as chemiluminescence that decay over time, rapid imaging is crucial. Existing systems, especially CCDs, often employ slow scanning modes that can cause signal attenuation during imaging, affecting quantitative accuracy.

[0009] System integration and intelligence: Most systems have limited functions and lack intelligent monitoring and management of samples, environment and equipment status, making it difficult to meet the needs of modern laboratory automation and intelligence.

[0010] Multimodal imaging capability: Different blotting methods (such as chemiluminescence, fluorescence, and colorimetry) may require different excitation or detection modes. Existing devices are usually optimized for a single mode, which is not flexible enough.

[0011] Therefore, there is an urgent need in this field for a bioimprint optical imaging system that can take into account high speed, high resolution, high sensitivity, high integration and multimodal imaging capabilities. Summary of the Invention

[0012] To address the shortcomings of existing technologies, this invention provides a rapid high-resolution bioimprint optical imaging system. This system innovatively integrates a deeply cooled sCMOS detector, an intelligent temperature-controlled pellet pressing module, a multispectral LED excitation source, an autofocus mechanism, and an embedded intelligent monitoring unit, aiming to achieve high-speed, high-resolution, high-sensitivity, multimodal, and intelligent imaging analysis of bioimprint samples.

[0013] To achieve the above objectives, the present invention is implemented through the following technical solution: a rapid high-resolution bioimprint optical imaging system, comprising a main control computer system, a high-precision mechanical motion platform, a multimodal optical imaging chamber, and an embedded system control core; The main control computer system is used to control the operation of the entire system, process image data, and provide a human-computer interaction interface. The high-precision mechanical motion platform includes an XY translation stage for carrying and positioning the sample and a Z-axis autofocus mechanism for achieving fast and accurate focusing. The multimodal optical imaging cabin includes: The sCMOS detector module uses a scientific-grade back-illuminated sCMOS sensor and integrates a multi-stage semiconductor cooling and liquid cooling heat dissipation system to control the sensor's operating temperature to -45°C and below. The intelligent temperature-controlled pressing module, located in front of the sCMOS detector photosensitive area, includes a light-transmitting pressing plate, heating element, temperature sensor and temperature controller, which can accurately control the temperature of the sample. A multispectral LED excitation source system, comprising an LED array with multiple independently controllable wavelength bands, is used to provide multicolor fluorescence excitation; The high-efficiency filter wheel module is located between the sCMOS detector and the pressing module, and is equipped with multiple filters that are automatically switched by a motor. The dark chamber shell and the light-blocking cover form a sealed light cavity; The embedded system control core is used to integrate control of motor movement, light source driving, temperature control, environmental monitoring, vibration sensing, positioning communication, and real-time image data preprocessing.

[0014] Preferably, the sCMOS detector module has an effective imaging area of ​​28mm×28mm, a pixel size of 6.5µm, and a resolution of 4.3 million pixels; the multi-stage semiconductor cooling and liquid cooling system includes at least two stages of semiconductor coolers connected in series and microchannel liquid cooling blocks coupled thereto.

[0015] Preferably, the light-transmitting plate of the intelligent temperature-controlled pressing module is made of quartz glass, the heating element is a transparent conductive heating film, the temperature control range is from room temperature to 70°C, and the control accuracy is ±0.5°C.

[0016] Preferably, the multispectral LED excitation light source system includes LED arrays in five bands: ultraviolet, blue, green, red, and visible light. Each array is equipped with a bandpass filter, and its switching and intensity can be independently programmed and controlled.

[0017] Preferably, the Z-axis autofocus mechanism uses a combination of a laser displacement sensor and a piezoelectric ceramic actuator to achieve closed-loop focus control.

[0018] Preferably, the embedded system control core is based on an ARM microprocessor and an FPGA, and the FPGA integrates hardware circuitry for real-time preprocessing algorithms to implement image flat field correction and dark background subtraction.

[0019] Preferably, the environmental monitoring integrated into the embedded system control core includes temperature and humidity monitoring within the imaging chamber; vibration sensing employs a MEMS accelerometer; and positioning and communication include a GPS / BeiDou module and a Wi-Fi / Bluetooth module.

[0020] Preferably, the sCMOS detector module communicates with the main control computer system via a Camera Link high-speed interface.

[0021] Preferably, the light-shielding cover is equipped with a safety interlock switch, which automatically shuts off the power supply to the multispectral LED excitation light source system when the light-shielding cover is opened. The method for bioimprint imaging using a rapid, high-resolution bioimprint optical imaging system is also disclosed, including the following steps: The imaging parameters are set through the main control computer system, including selecting the imaging mode, excitation wavelength, filter, exposure time, sample temperature and scanning area; The high-precision mechanical motion platform moves the sample stage to the initial imaging position, and the Z-axis autofocus mechanism performs autofocus. The embedded system control core controls the filter wheel to switch to the corresponding filter according to the settings. If it is in fluorescence mode, it turns on the LED light source of the specified wavelength and sets the intensity. If temperature control is enabled, it starts the intelligent temperature control pressing module to heat to the set temperature. The main control computer system triggers image acquisition, the sCMOS detector performs exposure under the control of the synchronization signal, and outputs the image data; After transmission and real-time preprocessing, the image data is displayed, stored, and analyzed in the main control computer system. Throughout the process, the embedded system control core continuously monitors environmental parameters and device status, and issues an alarm when an anomaly is detected.

[0022] This invention provides a rapid, high-resolution bioimprint optical imaging system. Compared with existing technologies, it has the following advantages: 1. Extremely high imaging sensitivity and signal-to-noise ratio: Deeply cooled sCMOS (-45°C) reduces dark current to an extremely low level. Combined with the high quantum efficiency (>95%) of the back-illuminated structure, it can detect extremely weak chemiluminescence or fluorescence signals. Intelligent temperature-controlled pressing further enhances signal output.

[0023] 2. Excellent spatial resolution: The small pixel size of 6.5µm combined with the high resolution of 4.3 million pixels can clearly distinguish closely adjacent protein bands or spots on the blot membrane, meeting the requirements of high-precision qualitative and quantitative analysis.

[0024] 3. Ultra-fast imaging speed: The inherent high-speed readout characteristics of sCMOS, combined with a high-speed interface, autofocus, and filter switching mechanism, enable full-field imaging time as short as milliseconds, effectively capturing the dynamic changes of chemiluminescence signals and avoiding quantitative errors caused by signal attenuation.

[0025] 4. Powerful multimodal imaging capabilities: It integrates multi-band excitation light sources and multi-channel filtering systems, and one set of equipment can complete imaging in multiple modes such as chemiluminescence, multicolor fluorescence, and colorimetry. It has a wide range of applications and high cost performance.

[0026] 5. High degree of intelligence and automation: It integrates automatic focusing, environmental monitoring, vibration sensing, wireless communication and real-time preprocessing functions, which reduces human intervention and errors, improves the repeatability and efficiency of experiments, and facilitates the information management of the laboratory.

[0027] 6. Excellent system integration and stability: The modular design tightly integrates optics, mechanics, electronics, and software, resulting in a compact structure and stable and reliable operation. Intelligent temperature control and liquid cooling systems ensure temperature stability during long-term exposures. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of the system of the present invention; Figure 2 This is a schematic diagram of the multimodal optical imaging chamber of the system of the present invention; Figure 3 This is a schematic diagram of the precision mechanical motion platform of the system of the present invention; Figure 4 This is an image showing the imaging effect of protein molecular weight standards using chemiluminescence mode in Embodiment 1 of the present invention; Figure 5 This is an imaging effect diagram of the simultaneous detection of two different target proteins using a dual-color fluorescence mode in Embodiment 2 of the present invention.

[0029] In the diagram: 1. Main control computer system; 2. High-precision mechanical motion platform; 21. XY translation stage; 22. Z-axis autofocus mechanism; 3. Multimodal optical imaging chamber; 31. Light shield; 32. Multispectral LED excitation light source system; 33. Intelligent temperature control pressing module; 34. High-efficiency filter wheel module; 35. sCMOS detector module; 36. Dark chamber shell; 4. Embedded system control core. Detailed Implementation

[0030] The technical solutions in the embodiments of the present invention have been clearly and completely described. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Please see Figure 1-3 This invention provides a technical solution: a rapid high-resolution bioimprint optical imaging system. The main control computer system 1 is specifically a main control computer workstation. This workstation is a computing platform equipped with a high-performance graphics processing unit, a built-in image acquisition card, a multi-axis motion control card, and dedicated image acquisition and control software. This software not only provides an intuitive user interface for setting all imaging parameters, controlling hardware actions, and displaying images in real time, but also integrates a powerful image post-processing and analysis toolkit, such as band density quantification, molecular weight calculation, and multi-channel fluorescence overlay analysis.

[0032] The high-precision mechanical motion platform 2 and the multimodal optical imaging cabin 3 are precisely integrated within a single system chassis. This chassis forms the mechanical and optical core of the system, providing structural support and light protection for its internal precision components.

[0033] The system's supporting electronic units, such as power management, motor drivers, temperature controllers, refrigerator power supplies, and safety protection circuits, are centrally located in an electrical control cabinet to achieve modular management and electromagnetic shielding.

[0034] In terms of connectivity: the main control computer workstation is connected to the sCMOS detector module 35 in the imaging chamber via a high-speed Camera Link cable to ensure lossless high-speed transmission of massive image data; at the same time, it communicates with the embedded system control core 4 in the electrical control cabinet and system host box via an integrated multi-functional control cable (such as using Ethernet or USB 3.0 interfaces) to transmit all control commands and receive status feedback.

[0035] The basic workflow of the system is as follows: The user places the prepared bioimprinted membrane (such as an NC membrane or PVDF membrane) sample on the sample stage of the main unit and selects a preset imaging method or custom parameters through computer software. Subsequently, the system automatically executes a series of actions: the mechanical platform moves the sample to the imaging start position, the autofocus mechanism precisely positions the focal plane, the filter is switched according to the imaging mode and the light source is set, exposure is initiated for image acquisition, data is transmitted and processed, and the results are displayed and stored. The entire process is highly automated, minimizing human error.

[0036] The multimodal optical imaging chamber 3 is a strictly light-shielded sealed cavity, consisting of a dark chamber shell 36 and a light-shielding cover 31 with a sealing strip. A safety interlock switch (not shown) is installed inside the light-shielding cover 31. When it is opened, it will immediately cut off the power supply to the excitation light source to prevent strong light from causing potential damage to the operator's eyes.

[0037] Inside the imaging chamber, the core components from top to bottom are as follows: Multispectral LED excitation light source system 32: It is uniformly distributed in a ring to ensure uniform illumination of the sample surface.

[0038] Intelligent temperature-controlled tablet pressing module 33: It is horizontally placed below the center of the light source ring, and the sample is placed directly on it for imaging.

[0039] High-efficiency filter wheel module 34: located directly below the tablet compression module.

[0040] sCMOS detector module 35: It is fixed to the bottom of the imaging chamber by a shockproof mounting bracket, with its photosensitive surface facing all the optical components above.

[0041] The high-precision mechanical motion platform 2 includes an XY translation stage 21 and a Z-axis autofocus mechanism 22, with the Z-axis autofocus mechanism 22 integrated on the imaging cabin base.

[0042] The sCMOS detector module 35 is key to achieving high-sensitivity, low-noise imaging. At its core is a scientific-grade back-illuminated sCMOS sensor chip with 4.3 million pixels (2048 x 2048) and a single pixel size of 6.5µm, ensuring a good balance between high resolution and high sensitivity. To drastically suppress thermal noise (dark current), the module employs a two-stage thermoelectric cooler (TEC) series configuration. The first-stage TEC is directly bonded to the back of the sensor package, while the second-stage TEC is coupled to the hot side of the first-stage TEC. The hot side of the second-stage TEC is in close contact with a carefully designed, high-efficiency liquid cooler block. The liquid cooler block has microchannels etched inside, efficiently removing heat through an external circulating coolant (typically a water-glycol mixture). The entire cooling assembly is encapsulated within a low-vacuum, insulated cavity, greatly reducing heat convection and conduction with the environment. A high-precision platinum resistance temperature sensor is embedded near the sensor to monitor the chip temperature in real time and feed it back to a dedicated PID temperature controller, ultimately maintaining the sensor chip's operating temperature stably at an extremely low level of -45°C ± 0.1°C. Image data is output at high speed via the CameraLink interface.

[0043] The intelligent temperature-controlled pellet pressing module 33 innovatively integrates sample temperature control into the imaging plane. Its core is an optical-grade quartz glass platen with extremely high transmittance and thermal stability. A transparent conductive tin oxide (ITO) heating film is deposited on the lower surface of the platen using magnetron sputtering. A miniature platinum resistance temperature sensor is precisely embedded in the edge of the platen, directly sensing the temperature of the platen surface (i.e., the sample contact surface). These components are connected to a high-precision PID temperature control circuit. Users can set the target temperature (ranging from room temperature to 70°C) in the software. The PID controller dynamically adjusts the power applied to the ITO heating film based on sensor feedback, achieving a temperature control accuracy of ±0.5°C. Uniform thermal conductivity and an insulating layer ensure temperature consistency throughout the imaging area. This function is crucial for accelerating chemiluminescence reaction kinetics and improving the quantum efficiency of certain fluorescent dyes (such as some protein dyes), enhancing signal intensity from the signal source.

[0044] To achieve flexible multicolor fluorescence imaging, a ring-shaped multispectral LED excitation light source system 32 and a high-efficiency filter wheel module 34 are designed.

[0045] It comprises five independent spectral channels: ultraviolet (center wavelength 365nm), blue (450nm), green (525nm), and red (630nm), as well as a white LED array (400-700nm) for visible light imaging. Each channel consists of a rigorously selected, wavelength-focused high-power LED array, equipped with sophisticated driving circuitry and a heat sink. Except for the white light channel, dedicated excitation filters are installed in front of the LED arrays in the other fluorescence excitation channels to further purify the excitation light and reduce stray light. The switching status and luminous intensity (0-100%) of all LEDs can be independently controlled by software.

[0046] The high-efficiency filter wheel module 34, located between the light source and the detector, is equipped with a porous filter wheel driven by a stepper motor. The filter wheel is pre-loaded with at least six filters, including a broadband pass filter (e.g., 400-700nm) for chemiluminescence imaging, emission filters corresponding to commonly used fluorescent dyes such as Cy2, Cy3, Cy5, and DAPI, and a dedicated transmission filter or neutral density filter for visible light imaging. Software controls the automatic rotation of the filter wheel to precisely insert the target filter into the optical path, ensuring that only emission light of specific wavelengths reaches the detector.

[0047] To ensure optimal focal plane acquisition for samples of varying thicknesses (such as imprinted films with pads of different thicknesses) or during multi-region scanning, the Z-axis autofocus mechanism 22 integrates an autofocus mechanism based on laser triangulation. A low-power laser displacement sensor emits a laser beam onto the sample surface and receives the reflected light. By calculating and accurately measuring the distance between the sample and the reference surface, this distance information is fed back to the control system in real time. This drives a piezoelectric ceramic actuator to fine-tune the position of the platform carrying the sCMOS detector module 35 in the Z-axis, achieving fast and accurate closed-loop focusing with a focusing repeatability accuracy within 1µm.

[0048] The system's "brain" is an embedded system control core based on an ARM microprocessor and FPGA, which achieves deep functional integration and intelligent collaboration. Motor drive interface: controls the servo motor of the XY translation stage 21, the stepper motor of the filter wheel, and the piezoelectric ceramic driver of the focusing mechanism.

[0049] Light source driving and synchronization unit: generates high-precision PWM signals to control the brightness of each LED and generates trigger pulses that are strictly synchronized with the sCMOS exposure signal to ensure "light-exposure" synchronization during fluorescence imaging and avoid photobleaching and signal crosstalk.

[0050] Temperature control unit: Simultaneously manages the cooling system of the sCMOS detector and the heating system of the intelligent tablet press, with independent closed-loop control for both, without interference between them.

[0051] Environmental sensor interface: Connects to temperature and humidity sensors installed in the imaging chamber to monitor and record environmental conditions in real time, providing environmental parameters to support the repeatability of experimental data.

[0052] Vibration monitoring unit: Continuously monitors equipment vibration using a highly sensitive MEMS accelerometer. Once the vibration amplitude exceeds a preset threshold (e.g., during long exposures), an alarm is immediately sent to the main control computer, and the current acquisition can be paused to protect image quality.

[0053] Positioning and Communication Module: Integrates a GPS / BeiDou dual-mode positioning chip to record the geographical location of the device (suitable for field or mobile laboratory scenarios); it is also equipped with Wi-Fi and Bluetooth modules to support wireless data transmission and remote monitoring of device status.

[0054] Data preprocessing unit: This is a hardware-level acceleration function implemented within the FPGA. Image data can undergo real-time flat-field correction (eliminating pixel response non-uniformity and illumination unevenness) and dark background subtraction (eliminating background noise) during transmission, greatly reducing the computational burden on the main control computer and improving overall processing efficiency.

[0055] Detailed explanation of system workflow The imaging system of this invention operates according to a highly automated and logically rigorous workflow. This workflow begins with simple user commands and ends with the output of high-quality image data; all complex operations in between are completed automatically by the system. The following will describe each stage in detail: Phase 1: Initialization and Parameter Setting System Power-On and Self-Test: The user turns on the power to the main control computer system 1, the host chassis, and the electrical control cabinet. The embedded system control core 4 starts up and executes the power-on self-test program, checking the following in sequence: Communication link: connection with the main control computer system 1, status of Camera Link and control bus.

[0056] Core module status: whether the cooling temperature of the sCMOS detector module 35 has reached the preset -45°C, the initial position of the high-efficiency filter wheel module 34, the limit switches of the XY translation stage 21, etc.

[0057] Safety interlock: Confirm that the light shield 31 is closed.

[0058] The self-test results are displayed in real time on the main control software interface. If any self-test fails, the system will display a clear alarm message and prevent subsequent operations to ensure operational safety.

[0059] Experimental parameter configuration: Users create or invoke an imaging experiment protocol within the dedicated software interface. The required parameters include: Imaging mode selection: Select "chemiluminescence", "fluorescence", "visible light (colorimetric method)" or a combination mode according to the sample labeling method.

[0060] Light source and filter (for fluorescence / visible light mode): Specify the corresponding excitation LED wavelength (such as blue light, green light, red light, white light) and its intensity (0-100%) for each detection channel, and specify the corresponding filter (emission filter or visible light special filter) on the filter wheel for each channel.

[0061] Exposure parameters: Set the exposure time (milliseconds to hours), gain, etc. of the sCMOS detector module 35.

[0062] Temperature control: Set the target temperature of the intelligent temperature control tablet pressing module 33 (such as room temperature 25°C or 37°C set to enhance the signal).

[0063] Scanning scheme: If the sample needs to be stitched together for imaging, then define the scanning area and movement path of the XY translation stage 21.

[0064] Phase Two: Sample Loading and Automatic Positioning Safe sample loading: The software prompts the user to open the light-shielding cover 31 to load the sample. The moment the light-shielding cover 31 is opened, the safety interlock switch immediately cuts off the power supply to the multispectral LED excitation light source system 32 to ensure safe operation. The user places the prepared imprint film sample flat on the quartz glass plate of the intelligent temperature-controlled pressing module 33, and then closes the light-shielding cover 31.

[0065] Autofocus: After the user clicks "Start Imaging," the system first executes the autofocus program. The Z-axis autofocus mechanism 22 is activated, and its laser displacement sensor emits a laser beam towards the sample surface and receives the reflected signal, accurately measuring the relative distance between the sample surface and the sensor. This distance data is fed back to the embedded system control core 4 in real time, which drives the piezoelectric ceramic actuator to fine-tune the Z-axis height of the sCMOS detector module 35 or the stage until the sample surface is precisely located on the optimal focal plane of the sCMOS detector. This process is a closed-loop control, ensuring both speed and accuracy of focusing.

[0066] Phase 3: Collaborative Imaging and Data Acquisition Environment preparation and module readiness: The PID temperature control circuit of the intelligent temperature control tablet pressing module 33 starts working. Based on the feedback from the platinum resistance temperature sensor, it precisely adjusts the power of the ITO heating film so that the sample can quickly reach and stabilize at the set temperature.

[0067] The stepper motor of the high-efficiency filter wheel module 34 receives the command and drives the filter wheel to rotate, so that the first filter specified in the protocol (such as a chemiluminescence broadband filter, an emission filter for a specific fluorescence channel, or a visible light imaging filter) is precisely inserted into the optical path.

[0068] Image acquisition execution: For chemiluminescence mode: all LED excitation sources remain off. The main control computer system 1 sends an exposure trigger signal to the sCMOS detector module 35, which performs exposure under deep cooling to capture the weak chemiluminescence signal generated by the sample itself.

[0069] For fluorescence imaging mode: the main control computer system 1 issues a synchronization pulse sequence that ensures the designated LED excitation source is synchronously lit at the moment the sCMOS detector module 35 begins exposure and precisely turned off at the end of exposure. This strict "light-exposure synchronization" mechanism minimizes photobleaching of the sample and guarantees signal authenticity.

[0070] For visible light imaging mode: The system activates the white LED array according to the settings to provide uniform broadband illumination and expose the sample for detecting visible light signals generated by colorimetric methods (such as HRP-DAB color development, ECL color development, or dye staining with Coomassie Brilliant Blue). For multi-channel fluorescence / combination mode: The system will automatically and quickly repeat the process of "switching filters, activating the corresponding LED, exposure, and deactivating LEDs" according to the protocol sequence, acquiring images of each channel sequentially.

[0071] Data Streaming and Real-Time Processing: The sCMOS detector module 35 converts the analog charge signal generated by exposure into digital image data, which is then transmitted at high speed to the main control computer via the Camera Link interface. Simultaneously, the FPGA data preprocessing unit within the embedded core works in parallel, performing flat-field correction (eliminating inconsistencies in detector pixel response and uneven illumination) and dark background subtraction (eliminating the background caused by dark current and readout noise) on the raw image data. This hardware-level real-time preprocessing significantly improves the uniformity and signal-to-noise ratio of the output image and reduces the post-processing burden on the main control computer.

[0072] Phase 4: Results Output and Status Monitoring Image display, storage, and analysis: High-quality pre-processed images are displayed in real-time on the software's main interface. The system automatically stores the image data along with all acquisition parameters (exposure time, temperature, filter position, etc.) on the hard drive. Users can immediately utilize the software's built-in analysis tools to perform operations such as band quantification, molecular weight analysis, multi-channel fluorescence overlay, and visible light image analysis.

[0073] Intelligent monitoring throughout the entire process: The embedded system control core 4 continuously runs in the background throughout all the above stages. Environmental monitoring: The temperature and humidity inside the imaging chamber are monitored in real time via an environmental sensor interface and recorded in the log.

[0074] Vibration monitoring: The MEMS accelerometer in the vibration monitoring unit continuously senses the vibration of the equipment. Once an abnormal vibration is detected (such as being hit during a long exposure), it will immediately trigger an interruption, suspend image acquisition and issue an alarm to protect data integrity.

[0075] Location and Communication: The location and communication module can record the device's geographical location and upload device status reports via wireless network.

[0076] Examples and Effects Demonstration Example 1: High-sensitivity chemiluminescence imaging Image was performed on a PVDF membrane transferred with complex cell lysate proteins and treated with HRP-labeled antibody and a highly sensitive chemiluminescent substrate. In the software, the "chemiluminescence" mode was selected, the filter wheel automatically switched to a wide-pass filter, and all excitation sources were turned off. The exposure time for the sCMOS detector module 35 was set to 3 minutes, and the pelleting temperature was set to 25°C (room temperature). Thanks to the extremely low background noise resulting from the deep cooling of the sCMOS to -45°C, the obtained image (see reference) is excellent. Figure 4 With a pitch-black background, the target protein bands are clear and sharp, and even very weak, low-abundance protein bands can be easily identified. The signal-to-noise ratio is much higher than that of conventional room-temperature CCD or CMOS systems.

[0077] Example 2: Fast Multicolor Fluorescence Imaging Image a Western blot membrane simultaneously incubated with FITC (green fluorescence) and Cy5 (far red fluorescence) labeled antibodies targeting different targets. Create a "multichannel fluorescence" protocol in the software: Channel 1: Select blue light excitation (450nm), Cy2 emission filter, exposure time 800ms.

[0078] Channel 2: Select red light excitation (630nm), Cy5 emission filter, exposure time 1.2s.

[0079] The system automatically completes the following steps within seconds: switching the filter wheel to channel 1, turning on the blue LED, exposure, turning off the light source, switching the filter wheel to channel 2, turning on the red LED, and exposure. The entire process is fast and seamless. The final images of the two channels are shown in the reference image. Figure 5The software can be used to synthesize pseudo-colors (such as green and red) to visually display the expression and co-localization of two target proteins in the sample, enabling multiple detection on the same membrane, which is efficient and avoids the errors of multiple experiments.

[0080] Example 3: Visible Light (Colorimetric) Imaging Imaging was performed on an immunohistochemical (IHC) slide treated with DAB (3,3'-diaminobenzidine) or a protein gel stained with Coomassie Brilliant Blue. In the software, the "Visible Light Imaging" mode was selected, the filter wheel was switched to a dedicated visible light filter, and the white LED array was activated to provide uniform illumination. An appropriate exposure time was set (e.g., 100ms), and the intelligent temperature-controlled pelleting module 33 was set to room temperature. Due to the uniform white light illumination and the high dynamic range of the sCMOS detector, the obtained visible light image exhibited accurate color reproduction, high contrast in bands or stained areas, and rich detail, making it directly usable for density-based quantitative analysis. Example 4: Intelligent Function Verification Vibration resistance: During a 10-minute chemiluminescence exposure, a simulated external impact caused vibration of the experimental stage. The MEMS accelerometer within the system immediately detected the abnormal vibration signal. The embedded core sent an interrupt signal to the main control software within milliseconds. The software interface displayed a warning and automatically paused the exposure. It resumed after the vibration stopped and the user confirmed, effectively preventing image blurring and failure.

[0081] Precise temperature control application: In an experiment studying a temperature-sensitive fluorescent dye, the tableting temperature was precisely set and stabilized at 42.0°C. The PID temperature control system, through real-time feedback from a platinum resistance sensor, kept temperature fluctuations within ±0.3°C, ensuring that the fluorescence signal was detected at the optimal reaction temperature and improving the reliability of quantitative data.

[0082] Equipment Management and Traceability: In research institutes that require strict asset management, the system regularly reports the location of equipment via GPS module and sends key usage logs (such as power-on time, cumulative imaging count, and alarm records) to the central server via Wi-Fi, realizing digital management and usage traceability of equipment.

[0083] In summary, this invention has successfully created a bioimprint optical imaging platform that excels in imaging speed, spatial resolution, detection sensitivity, functional completeness (covering chemiluminescence, multicolor fluorescence, and visible light imaging), and intelligence by deeply integrating and optimizing multiple advanced technologies such as deep-cooled sCMOS technology, intelligent sample temperature control, multispectral excitation (including white light and visible light channels), high-speed automatic filtering, laser autofocus, and embedded intelligent monitoring. This provides a powerful tool for life science research and clinical diagnosis.

[0084] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0085] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A rapid, high-resolution bioimprint optical imaging system, characterized in that: It includes a main control computer system, a high-precision mechanical motion platform, a multimodal optical imaging cabin, and an embedded system control core; The main control computer system is used to control the operation of the entire system, process image data, and provide a human-computer interaction interface. The high-precision mechanical motion platform includes an XY translation stage for carrying and positioning the sample and a Z-axis autofocus mechanism for achieving fast and accurate focusing. The multimodal optical imaging cabin includes: an sCMOS detector module, which adopts a back-illuminated sCMOS sensor and integrates a multi-stage semiconductor cooling and liquid cooling heat dissipation system to control the sensor's operating temperature; The intelligent temperature-controlled pressing module, located in front of the sCMOS detector photosensitive area, includes a light-transmitting pressing plate, heating element, temperature sensor and temperature controller, which can accurately control the temperature of the sample. A multispectral LED excitation source system, comprising an LED array with multiple independently controllable wavelength bands, is used to provide multicolor fluorescence excitation; The high-efficiency filter wheel module is located between the sCMOS detector and the pressing module, and is equipped with multiple filters that are automatically switched by a motor. The dark chamber shell and the light-blocking cover form a sealed light cavity; The embedded system control core is used to integrate control of motor motion, light source drive, temperature control, environmental monitoring, vibration sensing, positioning communication, and real-time image data preprocessing.

2. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The multi-stage semiconductor cooling and liquid cooling system includes at least two semiconductor coolers connected in series and a microchannel liquid cooling block coupled thereto.

3. The bioimprint optical imaging system according to claim 1, characterized in that: The light-transmitting plate of the intelligent temperature-controlled pressing module is made of quartz glass, and the heating element is a transparent conductive heating film. The temperature control range is from room temperature to 60-80°C.

4. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The multispectral LED excitation light source system includes LED arrays in five bands: ultraviolet, blue, green, red, and visible light. Each array is equipped with a bandpass filter, and its switching and intensity can be independently programmed and controlled.

5. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The Z-axis autofocus mechanism uses a combination of a laser displacement sensor and a piezoelectric ceramic actuator to achieve closed-loop focus control.

6. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The embedded system control core is based on an ARM microprocessor and an FPGA. The FPGA integrates hardware circuitry for real-time preprocessing algorithms to implement image flat field correction and dark background subtraction.

7. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The embedded system control core integrates environmental monitoring including temperature and humidity monitoring inside the imaging chamber; vibration sensing uses a MEMS accelerometer; and positioning and communication includes GPS, Beidou module, Wi-Fi and Bluetooth module.

8. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The sCMOS detector module communicates with the main control computer system via the Camera Link high-speed interface.

9. The rapid high-resolution bioimprint optical imaging system according to claim 1, characterized in that: The light-shielding cover is equipped with a safety interlock switch, which automatically shuts off the power supply to the multispectral LED excitation light source system when the light-shielding cover is opened.

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