Microscope synchronous control system and method

Abstract

The present application relates to the field of microscope imaging control, in particular to a kind of microscope synchronous control system and method, system includes: microscope high-speed synchronous control device, camera, laser, LED light source and spatial light modulator;The host computer is connected with the microscope high-speed synchronous control device by serial port, for sending specific setting sequence and data sequence to the microscope high-speed synchronous control device Setting or sending trigger sequence to control the microscope high-speed synchronous control device generates digital signal.The synchronous operation of the camera, laser, LED light source and spatial light modulator and analog signal control piezoelectric ceramic Z table is realized in complex microscope system according to the digital signal controlled by the microscope high-speed synchronous control device Precise shooting of camera.

Description

Technical Field

[0001] The present invention relates to the field of microscope imaging control, and in particular to a microscope synchronization control system and method. Background Art

[0002] Microscope imaging is widely used in biological research and is a crucial tool for uncovering life processes. The core of microscope imaging control systems is multi-dimensional image acquisition (including time t, XY position, Z-axis slices, and multiple fluorescence channels). To enable the microscope to execute imaging tasks according to a predetermined workflow, this is traditionally accomplished by sending commands from a computer to sequentially modify parameters or issue action instructions for each device. This inevitably requires software-based communication with each device, which often takes several milliseconds, resulting in additional delays (ranging from tens of milliseconds) for each frame captured. Furthermore, in traditional methods, the time sequence required for camera capture is typically controlled by a software-controlled clock sequence generated by the computer. Due to the non-real-time nature of the operating system, this time sequence can fluctuate by several to tens of milliseconds. For high-speed microscopy imaging, such as studying bacterial motility, which requires acquisition speeds of 30 Hz or higher, precise timing control cannot be achieved through computer software, requiring the use of a TTL external trigger signal to control the microscope's capture.

[0003] Improving the data acquisition efficiency of microscope images is of great significance to scientific research. It can 1) increase the capacity of scientific datasets and enhance the statistical reliability of results; and 2) shorten the time required to acquire the same data, improving the timeliness of measurements. Reducing unnecessary latency is a fundamental approach to improving acquisition efficiency. This requires the microscope system to achieve high-speed and high-precision control of the camera, motorized stage, and light source in both time and space. This is achieved by: The camera is immediately exposed after the motorized stage moves into position, and the light source of the corresponding wavelength is simultaneously turned on. Once the camera exposure is complete, the next stage movement begins immediately. This cycle achieves low latency in camera image acquisition in time (t) and space (x, y, z). For fluorescence imaging, delayed activation or premature deactivation of the excitation light source within the camera exposure time can weaken fluorescence excitation. Premature activation or delayed deactivation of the excitation light source can increase toxicity and photobleaching in biological samples. High synchronization between the camera imaging time and the excitation light source activation time is a prerequisite for achieving long-term and high-frequency fluorescence image acquisition. Traditional software control methods cannot meet such high-precision timing requirements, so there is an urgent need for a hardware control device that can achieve high-precision and high-speed image acquisition, which is of great significance for high-speed and high-throughput image acquisition of fluorescence microscopes. Summary of the Invention

[0004] The embodiment of the present invention provides a microscope synchronization control system and method, the system has multiple digital signal outputs, and realizes accurate shooting of complex processes in the system.

[0005] According to one embodiment of the present invention, there is provided a microscope synchronization control system, comprising: a host computer, a microscope high-speed synchronization control device, a camera, a laser, an LED light source, and a spatial light modulator;

[0006] The host computer is connected to the microscope high-speed synchronous control device through a serial port, and is used to send a specific setting sequence and data sequence to set the microscope high-speed synchronous control device or send a trigger sequence to control the microscope high-speed synchronous control device to generate a digital signal;

[0007] The microscope high-speed synchronization control device controls the synchronous operation of the camera, laser, LED light source and spatial light modulator according to digital signals. The digital signals include camera exposure trigger signal, LED light source trigger signal, laser trigger signal and spatial light modulator trigger signal.

[0008] The microscope high-speed synchronous control device sends a camera shooting trigger signal to the camera to control the camera to shoot; the microscope high-speed synchronous control device sends a digital signal to the LED power supply to control the LED power supply to turn on;

[0009] The microscope high-speed synchronization control device sends a laser trigger signal to the laser, and the microscope high-speed synchronization control device sends a modulator trigger signal to the spatial light modulator. When the camera is shooting, the spatial light modulator is triggered, and the spatial light modulator outputs a TTL signal to the microscope high-speed synchronization control device. The microscope high-speed synchronization control device sends the laser trigger signal and the spatial light modulator trigger signal as "AND" to the laser and spatial modulator to synchronize the input signals of the spatial light modulator and the laser, thereby realizing precise timing control of the LED light source and the camera, and enabling the camera to shoot.

[0010] Furthermore, the system also includes a motorized translation stage, and the digital signal also includes a motorized translation stage trigger signal. The microscope high-speed synchronous control device sends the motorized translation stage trigger signal to the motorized XY translation stage, and the motorized translation stage moves in the XY two-dimensional plane according to the received motorized translation stage trigger signal.

[0011] After the camera finishes shooting, the microscope high-speed synchronization control device will send a trigger signal of the motorized translation stage to the motorized translation stage, causing the motorized translation stage to move to the next preset position;

[0012] After the motorized translation stage moves to the preset position, an external trigger signal is sent to the microscope's high-speed synchronization control device, causing it to start the next shooting cycle.

[0013] Furthermore, the system also includes a piezoelectric ceramic Z stage, the digital signal also includes a Z stage analog signal, the microscope high-speed synchronous control device sends the Z stage analog signal to the piezoelectric ceramic Z stage, and the piezoelectric ceramic Z stage moves up and down according to the received Z stage analog signal;

[0014] After the camera shooting cycle ends, the piezoelectric ceramic Z stage will move up and down.

[0015] Furthermore, the microscope high-speed synchronous control device includes an integrated circuit board and an external power adapter. The integrated circuit board is provided with a main control chip, the main control chip is connected to two digital-analog chips, the two digital-analog chips are each connected to an operational amplifier filter, and the power adapter supplies power to the system; the power adapter provides the main control chip with a voltage converted by a transformer chip, and the power adapter provides digital-analog chip voltage to the two digital-analog chips and provides filtered voltage to the two operational amplifier filters;

[0016] The two op amp filters each output analog signals, and the main control chip provides sixteen-channel digital signal outputs.

[0017] Furthermore, the integrated circuit board is also provided with a host computer communication transcoding circuit, an electronic erasable programmable read-only memory circuit, a power supply isolation and voltage stabilization circuit, a digital-to-analog signal conversion circuit, a filtering and amplification circuit;

[0018] Host computer communication transcoding circuit, used to achieve two-way communication between the host computer and the microscope high-speed synchronization control device;

[0019] The electronic erasable programmable read-only memory circuit supports multiple rewrites and power-off storage, can pre-store data sequences in the storage module of the control box, and provides power-off storage function;

[0020] Power supply isolation and voltage stabilization circuit, used to isolate the input voltage and analog voltage, and filter the voltage;

[0021] Digital-to-analog signal conversion circuit, used to generate analog signals with 14-bit resolution to achieve high synchronization during microscope photography;

[0022] The filtering and amplifying circuit is used for amplifying the voltage and filtering the amplified voltage.

[0023] A microscope synchronization control method comprises the following steps:

[0024] The microscope high-speed synchronous control device receives the data sequence, and the microscope high-speed synchronous control device checks whether the data sequence complies with the specification;

[0025] If it meets the specifications, the data sequence is stored;

[0026] Determine whether the data sequence is in storage mode, if not, enter the waiting mode, if so, store the data sequence in the storage circuit in the microscope high-speed synchronization control device, and enter the waiting mode;

[0027] The microscope high-speed synchronous control device receives a trigger signal, the trigger carries at least a data sequence, and the microscope high-speed synchronous control device checks the data sequence;

[0028] Calling the data sequence, reading the data sequence in the microscope high-speed synchronous control device and caching it;

[0029] The cached data sequence is called, and based on the data sequence, a digital signal is output. Based on the digital signal, precise timing control of the LED light source and the camera is achieved, so that the camera can take pictures.

[0030] Furthermore, before the microscope high-speed synchronization control device receives the trigger signal, the method further includes:

[0031] Set the camera's exposure time and other parameters.

[0032] Furthermore, after achieving precise timing control of the LED light source and the camera based on the digital signal so that the camera can take pictures, the following steps are also included:

[0033] The microscope's high-speed synchronization control device determines whether the shooting is completed;

[0034] If yes, the shooting is ended; otherwise, the camera is controlled to move on the XY two-dimensional plane and shoot until the shooting of the XY two-dimensional plane is completed.

[0035] Furthermore, after completing the shooting of the XY two-dimensional plane, the method further includes:

[0036] The microscope high-speed synchronous control device receives and controls the camera to move up or down to achieve shooting at another viewing distance until the shooting is completed.

[0037] Furthermore, before the microscope high-speed synchronization control device determines whether the shooting is completed, the method further includes:

[0038] Preset the number of times to shoot in each XY two-dimensional plane of the camera;

[0039] Preset the number of times the camera moves up or down, and the distance of each movement.

[0040] The microscope synchronization control system and method in the embodiments of the present invention include a high-speed synchronization control device for a microscope, a camera, a laser, an LED light source, and a spatial light modulator. A host computer is connected to the high-speed synchronization control device via a serial port, and is used to send specific setting sequences and data sequences to configure the high-speed synchronization control device, or to send trigger sequences to control the high-speed synchronization control device to generate digital signals. The high-speed synchronization control device controls the synchronous operation of the camera, laser, LED light source, and spatial light modulator based on the digital signals, enabling precise camera capture in complex microscope systems. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0042] Figure 1 This is a schematic diagram of the microscope synchronization control system of the present invention;

[0043] Figure 2 This is a signal output diagram of the microscope synchronization control system of the present invention;

[0044] Figure 3 The digital level change in a fast XY scan of the signal output of the synchronous control system of the microscope of the present invention;

[0045] Figure 4 The marked part is the present invention Figure 3 The first 220 milliseconds of the excerpt;

[0046] Figure 5 This is a schematic diagram of external interrupt microscope control in the microscope high-speed synchronous control system of the present invention;

[0047] Figure 6 Flowchart of the microscope synchronization control system method of the present invention;

[0048] Figure 7 This is the input process during the use of the microscope high-speed synchronous control system of the present invention;

[0049] Figure 8 This is the output process during the use of the microscope high-speed synchronous control system of the present invention;

[0050] Reference numerals: 1-host computer, 2-microscope high-speed synchronization control device, 3-camera, 4-laser, 5-LED light source, 6-spatial light modulator, 7-motorized translation stage, 8-voltage ceramic Z stage, 9-fluorescence filter block. DETAILED DESCRIPTION

[0051] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts should fall within the scope of protection of the present invention.

[0052] It should be noted that the terms "first", "second", etc. in the description and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that the numbers used in this way can be interchanged where appropriate, so that the embodiments of the present invention described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units that are not clearly listed or are inherent to these processes, methods, products or devices.

[0053] Example 1

[0054] According to an embodiment of the present invention, a method for liver segmentation in abdominal volume images based on deep learning is provided. Figure 1 , including: a host computer 1, a microscope high-speed synchronization control device 2, a camera 3, a laser 4, an LED light source 5 and a spatial light modulator 6;

[0055] The host computer 1 is connected to the microscope high-speed synchronous control device through a serial port, and is used to send a specific setting sequence and data sequence to set the microscope high-speed synchronous control device 2 or send a trigger sequence to control the microscope high-speed synchronous control device 2 to generate a digital signal;

[0056] The microscope high-speed synchronization control device 2 controls the synchronous operation of the camera 3, laser 4, LED light source 5 and spatial light modulator 6 according to the digital signal. The digital signal includes the exposure trigger signal of the camera 3, the trigger signal of the LED light source 5, the trigger signal of the laser 4 and the trigger signal of the spatial light modulator 6;

[0057] The microscope high-speed synchronous control device 2 sends a camera 3 shooting trigger signal to the camera 3 to control the camera 3 to shoot; the microscope high-speed synchronous control device 2 sends a digital signal to the LED power supply to control the LED power supply to turn on;

[0058] The microscope high-speed synchronization control device 2 sends a laser 4 trigger signal to the laser 4, and the microscope high-speed synchronization control device 2 sends a modulator trigger signal to the spatial light modulator 6. When the camera 3 is shooting, the spatial light modulator 6 is triggered, and the spatial light modulator 6 outputs a TTL signal to the microscope high-speed synchronization control device 2. The microscope high-speed synchronization control device 2 sends the laser 4 trigger signal and the spatial light modulator 6 trigger signal after "AND" to the laser 4 and the spatial modulator to synchronize the input signals of the spatial light modulator 6 and the laser 4, thereby realizing precise timing control of the LED light source 5 and the camera 3, and enabling the camera 3 to shoot.

[0059] The microscope synchronization control system and method in the embodiments of the present invention include a microscope high-speed synchronization control device 2, a camera 3, a laser 4, an LED light source 5, and a spatial light modulator 6. A host computer 1 is connected to the microscope high-speed synchronization control device via a serial port, and is used to send specific setting and data sequences to configure the microscope high-speed synchronization control device 2 or send trigger sequences to control the microscope high-speed synchronization control device 2 to generate digital signals. The microscope high-speed synchronization control device 2 controls the synchronous operation of the camera 3, laser 4, LED light source 5, and spatial light modulator 6 based on the digital signals, enabling precise capture by the camera 3 within a complex microscope system.

[0060] The main feature of existing microscopes in achieving high-speed synchronous shooting technology is that they use the timing of camera 3 as the main signal to trigger the synchronous control shooting of other devices. However, they are unable to perform high-precision timing control on other devices such as the motorized translation stage 7, and their support and scalability for the overall synchronization system of the microscope are insufficient. To address this shortcoming, the present invention designs and constructs a high-speed synchronous control device with multiple digital and analog output signals based on a high-precision timing control method triggered by the device externally, enabling precise shooting of complex processes in more complex microscope systems. In addition, by setting the external trigger of the synchronous control device itself, the microscope can complete a closed-loop, self-driven high-speed shooting process.

[0061] According to one aspect of an embodiment of the present invention, a high-speed synchronous control system for a microscope is provided, comprising a high-speed synchronous control device 2 for a microscope, a host computer 1, and microscope-related equipment, including a camera 3, a piezoelectric ceramic Z stage 8, a motorized XY translation stage (motorized translation stage 7), a laser 4, an LED light source 5, and a spatial light modulator 6. The motorized XY translation stage represents movement of the motorized translation stage 7 in the XY two-dimensional plane, while the piezoelectric ceramic Z stage 8 represents vertical movement in the Z-axis direction.

[0062] The microscope high-speed synchronization control device 2 includes an integrated circuit board and an external power adapter. The integrated circuit board contains the following components: a main control chip, peripheral circuits, a host computer 1 communication transcoding circuit, an electronically erasable programmable read-only memory circuit, a power supply isolation and voltage stabilization circuit, a digital-to-analog signal conversion circuit, and operational amplifier-related filtering and amplification circuits.

[0063] The host computer 1 is used for operator interaction, and sends specific setting sequences and data sequences through the serial port to set the microscope high-speed synchronization control device 2 or send trigger sequences to control the microscope high-speed synchronization control device 2 to generate signals. It can be but is not limited to laptop computers, desktop computers or single-chip microcomputers and other terminals that can communicate.

[0064] Microscope-related equipment includes a camera 3, a piezoelectric ceramic Z-stage 8, a motorized translation stage 7, a laser 4, an LED light source 5, and a spatial light modulator 6. The exposure mode of camera 3 is generally set by each manufacturer, with external triggering being the basic mode, where exposure of camera 3 is triggered by a pulse level. The piezoelectric ceramic Z-stage 8 can be controlled by an analog voltage, with the voltage range and step size determined by travel and accuracy. The motorized translation stage 7 has two signals: one as the stage's input signal, whose pulse level triggers the stage's movement, and one as the stage's output signal, which transmits a pulse level after the stage is in position. The switching of the laser 4 and LED light source 5 is triggered by a high-level pulse. Furthermore, because the input signal of the spatial light modulator 6 has high timing requirements, it is generally controlled by a high-precision TTL level. Users can choose different input and output modes. The microscope high-speed synchronous control system of the present invention serves as the signal control center, synchronizing the input signals of the spatial light modulator 6 and the laser 4 to achieve high-precision timing control of the light source and camera 3.

[0065] refer to Figure 1 and Figure 2 In this application, the communication between the host computer 1 and the microscope high-speed synchronous control device 2, and the signal transmission between the microscope high-speed synchronous control device 2 and the related equipment of the microscope are specifically:

[0066] Communication between the host computer 1 and the microscope high-speed synchronous control device 2 consists of an input sequence sent by the host computer 1 to the microscope high-speed synchronous control device 2 and a palindrome sequence fed back by the microscope high-speed synchronous control device 2. The input sequence includes a setup sequence for setting the operating mode of the microscope high-speed synchronous control device 2, such as turning on or off external interrupts and storage functions; and data sequences and signal trigger sequences for different modes, which describe the signal sequences that the microscope high-speed synchronous control device 2 needs to send. The palindrome sequence includes data from the microscope high-speed synchronous control device 2, system errors, and palindromes indicating the completion of the current task.

[0067] The signal transmission between the microscope high-speed synchronous control device 2 and microscope-related equipment can be summarized as the microscope high-speed synchronous control device 2 providing no less than sixteen digital signal outputs, no less than two analog outputs, and no less than two interrupt inputs to other microscope-related equipment; Figure 2 Digital signal 1 to digital signal 16 correspond to the output of the first to sixteenth digital signals. The digital signal output is used to control TTL trigger devices such as camera 3, electric translation stage 7, laser 4, LED light source 5, spatial light modulator 6, etc. In this example, the analog output is used to control the piezoelectric ceramic Z stage 8, with an effective range of 0 to 10V and a step size of 1mV. The interrupt input is used for external TTL pulse signals to trigger or influence the signal transmission of this device, so as to achieve precise synchronization control with microscope-related equipment. External interrupts can also support other complex signal input modes, such as the output signal of the 68-bit image of the spatial light modulator, and the output after performing "and" and "and" calculations with the specified digital signal, so as to achieve precise synchronization with the multi-channel light source signal when the spatial light modulator 6 displays an 8-bit image.

[0068] The microscope high-speed synchronous control device 2 includes an integrated circuit board and an external power adapter. A main control chip is provided on the integrated circuit board. The main control chip is connected to two digital-analog chips. The two digital-analog chips are each connected to an operational amplifier filter. The power adapter powers the system. The power adapter provides the main control chip with the voltage converted by the transformer chip. The power adapter provides digital-analog chip voltage for the two digital-analog chips and provides filtering voltage for the two operational amplifier filters. The two operational amplifier filters each output an analog signal, and the main control chip provides sixteen-channel digital signal output.

[0069] Specifically, the integrated circuit board in the microscope high-speed synchronous control device 2 mentioned in the present invention includes a main control chip, peripheral circuits, a host computer 1 communication transcoding circuit, an electronic erasable programmable read-only memory circuit, a power supply isolation and voltage stabilization circuit, a digital-to-analog signal conversion circuit, and operational amplifier-related filtering and amplification circuits. The host computer 1 communication transcoding circuit is used to achieve two-way communication between the host computer 1 and the microscope high-speed synchronous control device 2; the electronic erasable programmable read-only memory circuit has the functions of supporting multiple rewrites and power-off storage, enabling the pre-storage of data sequences into the storage module of the control box and providing power-off storage; the power supply isolation and voltage stabilization circuit is used to isolate the input voltage from the analog voltage and filter the voltage; the digital-to-analog signal conversion circuit is used to generate a 14-bit resolution analog signal to achieve high synchronization during microscope imaging; and the filtering and amplification circuit is used to amplify the voltage and filter the amplified voltage.

[0070] In this example, the main control chip and peripheral circuits use an STM32H7 series chip with a main frequency of up to 480MHz. Its peripheral circuits include a crystal oscillator circuit, a programming circuit, and a manual reset circuit. Its high main frequency allows for high-precision timing control, achieving high synchronization during microscope imaging and maximizing the device's performance.

[0071] In this example, host computer 1 uses USB-to-serial communication. This method simplifies communication, requiring only two transmission lines, one for transmitting and one for receiving, to achieve bidirectional communication. The transcoding circuit uses the commonly used serial port converter chip CH340, which can support high baud rate serial communication.

[0072] In this example, the electronically erasable programmable read-only memory circuit uses the AT24 series chip, which has 524,288 bits of storage space and supports multiple rewrites and power-off storage. This allows pre-storing data sequences into the control box's storage module and provides power-off storage.

[0073] In this example, the power supply isolation and voltage regulation circuit uses the TPH1515S-3W chip to isolate the input voltage and analog voltage, and uses the LM1117S, TPS5430, and TPS7A330 to obtain the digital 5V, digital 3.3V, analog 5V, analog 12V, and analog -12V voltages required by the circuit, and uses corresponding methods to filter the voltage.

[0074] In this example, the digital-to-analog signal conversion circuit uses the DAC904 high-speed digital-to-analog converter chip to generate 14-bit analog signals. Its 165MSPS conversion rate enables high-precision timing control, achieving high synchronization during microscope imaging and maximizing the device's performance. The ADA4898 chip converts the DAC904's differential output signal into a single-ended signal.

[0075] In this example, the operational amplifier-related filtering and amplification circuits use the AD620 instrumentation amplifier chip as the amplification module, the AD810 as the active low-pass filter module, and the LM358 as the analog output range adjustment module. The analog output range of the microscope high-speed synchronization control device 2 is -11V to +11V, and in this example, the output range is adjusted to 0-10V.

[0076] The input voltage is provided by a separate 15V voltage regulator adapter. After passing through the DCDC transformer chip, the input voltage will be converted into: power supply voltage for the chip and USB serial port (digital 5V), analog signal digital-to-analog chip voltage (analog 5V) and filter voltage (analog 10V) after isolation and conversion. The main control chip communicates with the user's host computer terminal 1 through the USB serial port. User commands are sent to the main control chip in the form of hexadecimal encoding, and the information returned by the main control chip is read. The system can be set to external interrupt mode. After the external TTL signal enters, the chip's pre-stored signal output is triggered. In this example, the main control chip directly provides 16 digital signal outputs. The analog signal output is output by the main control chip after digital-to-analog conversion and filtering.

[0077] In this example, by connecting the signal to the external trigger pin of each device, the correspondence between each digital signal channel of the microscope high-speed synchronization control device 2 and the device function of the trigger channel is shown in Table 1. Table 1 shows the correspondence between digital signals and trigger device functions.

[0078]

[0079]

[0080] In the embodiment, the system includes a motorized translation stage 7, the digital signal includes a trigger signal of the motorized translation stage 7, the microscope high-speed synchronization control device 2 sends the trigger signal of the motorized translation stage 7 to the motorized XY translation stage, and the motorized translation stage 7 moves in the XY two-dimensional plane according to the received trigger signal of the motorized translation stage 7;

[0081] After the camera 3 finishes shooting, the microscope high-speed synchronization control device 2 will send a trigger signal of the electric translation stage 7 to the electric translation stage 7 to move the electric translation stage 7 to the next preset position;

[0082] After the electric translation stage 7 moves to the preset position, an external trigger signal is sent to the microscope high-speed synchronization control device 2 to start the next shooting cycle.

[0083] In the embodiment, the system further includes a piezoelectric ceramic Z stage 8, the digital signal includes a Z stage analog signal, the microscope high-speed synchronization control device 2 sends the Z stage analog signal to the piezoelectric ceramic Z stage 8, and the piezoelectric ceramic Z stage 8 moves up and down according to the received Z stage analog signal;

[0084] After the camera 3 shoots a cycle, the piezoelectric ceramic Z stage 8 moves up and down.

[0085] The following describes in detail the motion of the electric translation stage 7 and the piezoelectric ceramic Z stage 8 of the present invention using a specific embodiment:

[0086] refer to Figures 1 to 4In this example, the minimum time unit provided is 1 microsecond, and the minimum adjustable range is 0.25 microseconds. This application can be used to encode data sequences of any number of pins, any time length, and any level. Using the basic method requires specifying the required signal time sequence, that is, the user needs to clearly specify the level state before the change, the time point when the change occurs, and the level state after the change each time the digital or analog signal changes. Figure 3 and Figure 4 This image shows the sequence of several digital signals during a rapid XY scanning process. This rapid XY scanning process includes the synchronization of the exposure trigger signal from camera 3 with the trigger signals from LED light source 5 and laser 4, as well as the AND output of the signal from the micromirror high-speed synchronization control device and the feedback signal from spatial light modulator 6. Figure 3 and Figure 4 The Y-axis represents the level change of the digital signal, 1 represents the high level of the digital signal, and 0 represents the low level of the digital signal. The X-axis represents the time of system operation, in 0.1 milliseconds. Figure 4 The marked part is Figure 3 The first 220 milliseconds of the excerpt.

[0087] The rapid XY scanning capture process in this example follows the following steps: First, a rising edge trigger signal from camera 3 triggers the camera 3 exposure start (at 0.5 milliseconds on the timeline). From the moment camera 3 receives the rising edge signal until the hardware startup completes and capture begins, approximately 24 milliseconds are required. After the capture begins (at 23.5 milliseconds on the timeline), the digital signal corresponding to LED light source 5 is changed to a high level. The triggering of LED light source 5 is completely synchronized with the digital signal: when the digital signal is high, LED light source 5 turns on; when the digital signal is low, LED light source 5 turns off. In this example, the desired duration of LED light source 5 on is 50 milliseconds. At 73.5 milliseconds on the timeline, the digital signal goes low, LED light source 5 stops generating signals, and a single capture ends. In this example, the interval between multiple captures is set to 20 milliseconds, with the next capture starting at 93.5 milliseconds. The operation is identical to the previous shot. The rising edge of the trigger signal from camera 3 triggers the camera 3 exposure (at 95 milliseconds on the timeline). After the start of the shot (at 119 milliseconds on the timeline), the spatial light modulator 6 (in this example, the second digital signal) and the corresponding digital signals from laser 4 (in this example, the fourth, fifth, sixth, and seventh digital signals) are triggered. Simultaneously, when spatial light modulator 6 is triggered, its output, a specially modulated TTL signal, is fed back to the microscope's high-speed synchronization control device 2 as an external interrupt signal. The digital signal sent by the microscope's high-speed synchronization control device 2 to laser 4 is the output of the AND operation between the set laser 4 signal and the spatial light modulator 6 signal acting as the external interrupt. The subsequent shots involving laser 4 follow the same procedure. This ensures that the laser's on / off timing is fully synchronized with the flipping timing of the liquid crystal in spatial light modulator 6. After a shooting cycle including the signals of the LED light source 5 and the four lasers 4 is completed, the microscope high-speed synchronous control device 2 will send a trigger signal of the electric translation stage 7 to complete the shooting, so that the electric translation stage 7 moves to the next field of view. After the electric translation stage 7 finishes moving, an external trigger signal will be sent to the microscope high-speed synchronous control device 2 to start the next shooting cycle.

[0088] In Z-scan mode, the piezoelectric Z-stage performs a single Z-axis movement after each capture cycle. In this example, 17 images were captured, starting at -1.6nm from the piezoelectric Z-stage's relative center position (75nm was chosen as the center position for this example, with a total travel of 150nm). The Z-axis increments were 0.2nm after each capture cycle. Table 2 shows the changes in digital and analog signals and the data transmitted during a single XY scan.

[0089]

[0090] Table 2

[0091] Following the above operation, the digital and analog signals will change levels at the following times: 0 ms, 0.5 ms, 10.5 ms, 23.5 ms, 74.5 ms, and so on. At each time point, a set of data is required to describe the changed digital and analog signals, as well as the duration of the change. As shown in the table above, at 0 ms, the digital signals are all low, Analog Signal 1 is at 4.8933V, corresponding to a piezoelectric Z-stage height of 73.4nm, and Analog Signal 2 is a constant 5V. At 0.5 ms, the trigger signal for camera 3 (digital signal 1) goes high, converting its digital signal to hexadecimal 0x0001. Analog Signal 1 remains at 4.8933V, and Analog Signal 2 remains at a constant 5V. After completing one shooting cycle, the second shooting cycle begins at 93.5ms. The digital signal level at this time is the same as when it was 0.5V, but the analog signal level has stepped by 0.2nm, and its level is 4.9067V. The analog signal 2 remains constant at 5V.

[0092] The above basic control method can describe all output sequences. However, for some situations where the duration is long and the sequence content is relatively repetitive, some general complex methods can reduce code redundancy, improve the efficiency of the single-chip microcomputer processing, and reduce the time it takes for the single-chip microcomputer to process the data sequence. For example, for completely repetitive sequences, a description of the number of cycles can be added to the sequence, which can achieve a specific output signal triggered at a specific frequency. In addition, based on user needs, specific data sequences can be customized to maximize the efficiency of sending sequences. The microscope high-speed synchronous control device 2 currently includes the classification of data sequences in Table 3:

[0093]

[0094] Table 3

[0095] The control method of microscope high-speed synchronous control system is as follows Figure 5As shown. In this example, the shooting process of the microscope imaging camera 3 is first set by the PC end. The camera 3 and the light source intensity and other parameters are set to a waiting state according to the trigger mode of the relevant equipment. In the example of the present invention, in the XY scanning mode, the microscope uses the MS2000 electric translation stage 7 of ASI as the XY electric translation stage 7, loads the fast XY firmware with ARRAY scanning, the laser 4 uses the OBIS series laser of Coherent, the LED uses the LED of Thorlabs or the PE4000 series of CoolLED, and the spatial light modulator 6 uses the QXGA-R10 series chip of ForthDD. The TTL level of the microscope high-speed synchronous control device 2 is used as a pulse signal to trigger the movement of the XY electric translation stage 7. After the electric translation stage 7 moves into position, it sends a TTL pulse signal as an external interrupt of the control system, triggering the control system to send the timing that has been written into the storage, realize the exposure of the camera 3, light source and other equipment, and complete the shooting of the microscope image. The signal of the electric translation stage 7 moving into position is used as the trigger of the next signal, and the pre-stored sequence is used to improve the operating efficiency of the equipment. In Z-scan mode, the piezoelectric ceramic Z stage 8 is controlled by a 0-10V analog voltage, sending a capture sequence within each analog signal step to complete the microscope image capture of each channel. In summary, multi-dimensional microscope capture of time, XY and Z scanning and channels is achieved.

[0096] refer to Figure 7 and Figure 8 , describes the two main conventional processes of interaction between the microscope high-speed synchronous control device 2 in the system, Figure 7 This paper describes the input process for using a microscope's high-speed synchronous control system, primarily involving the interaction between a host computer 1 and a microscope's high-speed synchronous control device 2. Users can edit data commands on host computer 1 and generate a visual sequence for verification using specialized software. After sending data commands to the microscope's high-speed synchronous control device 2, the device itself verifies the commands and provides feedback. In certain high-speed continuous output scenarios, the verification and feedback phases can be disabled to increase operating speed. If the verification sequence is correct, the data commands are stored in a cache, and the main control chip determines whether to store them in the electronically erasable programmable read-only memory circuit, entering standby mode.

[0097] Figure 8 This paper describes the general output process of the microscope high-speed synchronous control system, which mainly includes the interaction between the host computer 1, the microscope high-speed synchronous control device 2 and the microscope peripherals. Usually, the user needs to set up the microscope peripherals first, such as Figure 7 and Figure 8As shown, the user needs to set the exposure time and other parameters of the camera 3 on the host computer 1. After the settings are made, the trigger phase can begin. There are currently two ways to trigger the output signal. The first is to use the host computer 1 to directly send a trigger instruction, and the second is for the peripheral to send an external interrupt signal after the preparation is completed. The second method can relatively provide a faster trigger speed. After receiving the instruction, the main control chip determines whether to read the data in the electronic erasable programmable read-only memory circuit. Figure 8 After a capture cycle completes, the microscope high-speed synchronous control device 2 determines whether all operations are complete. If so, it issues a trigger signal to the motorized stage 7 indicating that the capture is complete, causing the stage 7 to move to the next field of view. After the stage 7 completes its movement, it sends an external trigger signal to the microscope high-speed synchronous control device 2 to initiate the next capture cycle. If all operations are complete, the cycle ends and the system awaits subsequent instructions.

[0098] like Figure 3 The following figure shows the signal output of multiple signals in a fast XY scan. The data instructions in the basic mode are:

[0099]

[0100] Every ten bits of data are required when a level changes. The first four bits are the hexadecimal digits represented by the digital signal after the current level change, and the last six bits represent the time until the next level change.

[0101] Example 2

[0102] According to an embodiment of the present invention, a microscope synchronization control method is provided. Figure 6 , including the following steps:

[0103] S101: The microscope high-speed synchronous control device receives a data sequence, and the microscope high-speed synchronous control device checks whether the data sequence meets the specification;

[0104] S102: If the data meets the specifications, the data sequence is stored;

[0105] S103: Determine whether the data sequence is in storage mode, if not, enter the waiting mode, if so, store the data sequence in the storage circuit in the microscope high-speed synchronization control device, and enter the waiting mode;

[0106] S104: The microscope high-speed synchronous control device receives a trigger signal, the trigger carries at least a data sequence, and the microscope high-speed synchronous control device checks the data sequence;

[0107] S105: calling the data sequence, reading the data sequence in the microscope high-speed synchronous control device and caching it;

[0108] S106: calling the cached data sequence, outputting a digital signal based on the data sequence, and achieving precise timing control of the LED light source and the camera based on the digital signal, so that the camera can take pictures.

[0109] This application uses a microscope high-speed synchronization control device to output digital signals based on data sequences to control the camera's precise shooting in a complex microscope system.

[0110] The microscope synchronization control method of the present invention is described in detail below with reference to specific embodiments:

[0111] Step 1: The host computer 1 and the microscope high-speed synchronous control device 2 interact with each other, and the user edits the data instructions carrying the data sequence on the host computer 1 and sends them to the microscope high-speed synchronous control device 2; the user edits the data instructions on the host computer 1 and can use specific software to generate a visual sequence for verification; in addition, the microscope high-speed synchronous control device 2 itself can also check whether the data sequence meets the specifications and can be correctly identified, and provide feedback. In the case of certain high-speed continuous output, the verification and feedback stages can be shielded to achieve the purpose of improving the operating speed.

[0112] Step 2: If the data sequence is verified to be in compliance with the specification or the sequence is correct, the data sequence will be stored in the cache.

[0113] Step 3: The main control chip determines whether the data sequence is to be stored in the electronic erasable programmable read-only memory circuit. If not, it enters the waiting mode. Otherwise, it stores the data sequence in the storage circuit in the microscope high-speed synchronization control device 2 and enters the waiting mode.

[0114] The above steps 1 to 3 are the input process of the microscope high-speed synchronous control system. Figure 7 shown.

[0115] Step 4: The microscope high-speed synchronous control device 2 receives a trigger signal, which carries at least a data sequence, and the microscope high-speed synchronous control device 2 checks the data sequence;

[0116] Before the microscope high-speed synchronous control device 2 receives the trigger signal, the method further includes:

[0117] Set the exposure time and other parameters of camera 3.

[0118] Specifically, the interaction between the microscope high-speed synchronization control device 2 and the microscope peripherals. Generally, the user needs to set the microscope peripherals first, and set the exposure time and other parameters of the camera 3 on the host computer 1. After the settings are made, the triggering phase can begin.

[0119] There are currently two ways to trigger the output signal. The first is to use the host computer 1 to directly send a trigger instruction, and the second is for the peripheral to send an external interrupt signal after preparation is completed. The second method can relatively provide a faster trigger speed.

[0120] Step 5: After receiving the trigger signal, the main control chip of the microscope high-speed synchronous control device 2 determines whether to read the data in the electronic erasable programmable read-only memory circuit. If the data sequence is called, the data sequence in the microscope high-speed synchronous control device 2 is read and cached.

[0121] Step 6: Call the cached data sequence, and based on the data sequence, output digital signals to the camera 3, the piezoelectric ceramic Z stage 8, the motorized XY translation stage, the LED light source 5, and the spatial light modulator 6, so that the timing of the LED light source 5 and the camera 3 is precisely controlled, so that the camera 3 can take pictures.

[0122] The above steps 4 to 6 are the output process of the microscope high-speed synchronous control system. Figure 8 shown.

[0123] In the embodiment, after the precise timing control of the LED light source 5 and the camera 3 is realized based on the digital signal so that the camera 3 takes pictures, the following steps are further included:

[0124] The microscope high-speed synchronization control device 2 determines whether the shooting is completed;

[0125] If so, the shooting is ended; otherwise, the camera 3 is controlled to move on the XY two-dimensional plane and shoot until the shooting of the XY two-dimensional plane is completed.

[0126] Specifically, after a capture cycle is complete, the microscope high-speed synchronous control device 2 determines whether the capture is complete. If not, the microscope high-speed synchronous control device 2 sends a trigger signal to the motorized translation stage 7 indicating that the capture is complete. Upon receiving the trigger signal, the motorized XY translation stage moves, causing the motorized translation stage 7 to move to the next field of view. After the motorized XY translation stage completes its movement, an external trigger signal is sent to the microscope high-speed synchronous control device 2 to initiate the next capture cycle. If all operations are complete, the cycle ends and the system awaits subsequent instructions.

[0127] In the embodiment, after completing the shooting of the XY two-dimensional plane, the method further includes:

[0128] The microscope high-speed synchronous control device 2 receives and controls the camera 3 to move up or down to achieve shooting at another viewing distance until the shooting is completed.

[0129] Before the microscope high-speed synchronization control device 2 determines whether the shooting is completed, the following steps are also included:

[0130] Preset the number of times camera 3 takes pictures on each XY two-dimensional plane;

[0131] The number of times the camera 3 moves up or down and the distance of each movement are preset.

[0132] In the microscope high-speed synchronous control system of the present invention, the TTL level of the microscope high-speed synchronous control device 2 is used as a pulse signal to trigger the movement of the XY motorized translation stage 7. After the motorized translation stage 7 moves into position, it sends a TTL pulse signal as an external interrupt of the control system, triggering the control system to send the stored timing sequence, realize the exposure of the camera 3, light source and other equipment, and complete the capture of the microscope image. The signal of the motorized translation stage 7 moving into position is used as the trigger for the next signal, and the pre-stored sequence is used to improve the operating efficiency of the equipment.

[0133] Compared with the existing technology, the present invention can solve the high synchronization problem in the current microscope shooting timing control and realize high-speed synchronous shooting. Its overall structure is simple, the cost is low, the actual operation is convenient, and the compatibility is wide. It is of great significance to improve the efficiency of microscope shooting and reduce the photobleaching during fluorescence imaging. Figure 1 As shown, the objective lens 10 observes through the fluorescence filter block 9, and the camera 3 takes pictures through the fluorescence filter block 9.

[0134] This invention has been tested and verified in actual on-device fluorescence microscope image capture, demonstrating its ability to capture multi-field, multi-channel fluorescence images. By editing the timing of the microscope capture protocol, the camera 3 exposure, light source activation, and spatial light modulator 6 synchronization are controlled. The invention boasts simple operation, wide applicability, and low overall cost, demonstrating its significant development prospects and commercial value.

[0135] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A microscope synchronization control system, characterized in that: include: Host computer, microscope high-speed synchronization control device, camera, laser, LED light source and spatial light modulator; The host computer is connected to the microscope high-speed synchronous control device via a serial port, and is used to send a specific setting sequence and data sequence to set the microscope high-speed synchronous control device or send a trigger sequence to control the microscope high-speed synchronous control device to generate a digital signal; The microscope high-speed synchronization control device controls the synchronous operation of the camera, laser, LED light source and spatial light modulator according to the digital signal and the analog signal to control the piezoelectric ceramic Z stage. The digital signal includes a camera exposure trigger signal, an LED light source trigger signal, a laser trigger signal and a spatial light modulator trigger signal. The analog signal controls the horizontal height of the piezoelectric ceramic Z stage. The microscope high-speed synchronous control device sends the camera shooting trigger signal to the camera to control the camera to shoot; the microscope high-speed synchronous control device sends a digital signal to the LED power supply to control the LED power supply to turn on; The microscope high-speed synchronization control device sends a laser trigger signal to the laser, and the microscope high-speed synchronization control device sends a modulator trigger signal to the spatial light modulator. When the camera takes pictures, the spatial light modulator is triggered. The spatial light modulator outputs a TTL signal to the microscope high-speed synchronization control device. The microscope high-speed synchronization control device sends the laser trigger signal and the spatial light modulator trigger signal as an "AND" signal to the laser and the spatial modulator to synchronize the input signals of the spatial light modulator and the laser, thereby achieving precise control of the timing of the LED light source and the camera, so that the camera takes pictures; The system further includes a motorized translation stage, and the digital signal further includes a motorized translation stage trigger signal. The microscope high-speed synchronous control device sends the motorized translation stage trigger signal to the motorized XY translation stage, and the motorized translation stage moves in the XY two-dimensional plane according to the received motorized translation stage trigger signal. After the camera finishes shooting, the microscope high-speed synchronous control device sends the motorized translation stage trigger signal indicating that the shooting is completed to the motorized translation stage, causing the motorized translation stage to move to the next preset position. After the motorized translation stage moves to the preset position, an external trigger signal is sent to the microscope high-speed synchronous control device, causing it to start the next shooting cycle. The system further includes a piezoelectric ceramic Z stage, and the digital signal further includes a Z stage analog signal. The microscope high-speed synchronization control device sends the Z stage analog signal to the piezoelectric ceramic Z stage, and the piezoelectric ceramic Z stage moves up and down according to the received Z stage analog signal. After the camera shooting cycle ends, the piezoelectric ceramic Z stage moves up and down. The microscope high-speed synchronous control device includes an integrated circuit board and an external power adapter. A main control chip is provided on the integrated circuit board. The main control chip is connected to two digital-analog chips. The two digital-analog chips are each connected to an operational amplifier filter. The power adapter powers the system; the power adapter provides the main control chip with the voltage converted by the transformer chip, and the power adapter provides the digital-analog chip voltage for the two digital-analog chips and provides the filtering voltage for the two operational amplifier filters; the two operational amplifier filters each output an analog signal, and the main control chip provides sixteen digital signal outputs.

2. The microscope synchronization control system according to claim 1, characterized in that: The integrated circuit board is also provided with a host computer communication transcoding circuit, an electronic erasable programmable read-only memory circuit, a power supply isolation and voltage stabilization circuit, a digital-to-analog signal conversion circuit, and a filtering and amplifying circuit; The host computer communication transcoding circuit is used to realize two-way communication between the host computer and the microscope high-speed synchronization control device; The electronic erasable programmable read-only memory circuit supports multiple rewrites and power-off storage, can pre-store data sequences in the storage module of the control box, and provides power-off storage function; Power supply isolation and voltage stabilization circuit, used to isolate the input voltage and analog voltage, and filter the voltage; The digital-to-analog signal conversion circuit is used to generate multiple independent 14-bit resolution analog signals to achieve high synchronization during microscope photography; The filtering and amplifying circuit is used for amplifying the voltage and filtering the amplified voltage.

3. A microscope synchronization control method, characterized in that: The following steps are involved: The microscope high-speed synchronous control device receives the data sequence, and the microscope high-speed synchronous control device checks whether the data sequence meets the specification; If it meets the specifications, the data sequence is stored; Determine whether the data sequence is in storage mode, if not, enter a waiting mode, if so, store the data sequence in a storage circuit in the microscope high-speed synchronization control device, and enter a waiting mode; The microscope high-speed synchronous control device receives a trigger signal, the trigger carries at least the data sequence, and the microscope high-speed synchronous control device checks the data sequence; Calling the data sequence, reading the data sequence in the microscope high-speed synchronous control device and caching it; The cached data sequence is called, and a digital signal is output based on the data sequence. Based on the digital signal, precise timing control of the LED light source and the camera is achieved, so that the camera can take pictures.

4. The microscope synchronization control method according to claim 3, characterized in that: Before the microscope high-speed synchronous control device receives a trigger signal, the method further includes: Set the exposure time parameters of the camera.

5. The microscope synchronization control method according to claim 4, characterized in that: After accurately controlling the timing of the LED light source and the camera based on the digital signal so that the camera can take pictures, the method further includes: The microscope high-speed synchronization control device determines whether the shooting is completed; If yes, the shooting is ended; otherwise, the camera is controlled to move on the XY two-dimensional plane and shoot until the shooting of the XY two-dimensional plane is completed.

6. The microscope synchronization control method according to claim 5, characterized in that: After completing the shooting of the XY two-dimensional plane, the method further includes: The microscope high-speed synchronous control device receives and controls the camera to move up or down to achieve shooting at another viewing distance until the shooting is completed.

7. The microscope synchronization control method according to claim 6, characterized in that: Before the microscope high-speed synchronization control device determines whether the shooting is completed, the method further includes: Preset the number of times the camera takes pictures on each XY two-dimensional plane; The number of times the camera moves up or down and the distance of each movement are preset.

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