Image-guided double-radio-frequency acousto-optic deflection laser cell cutting system and method

Abstract

The invention provides an image-guided double-radio-frequency acousto-optic deflection laser cell cutting system and method, and the system comprises a microscopic imaging module which is used for collecting a cell image; the imaging control module is used for controlling the microscopic imaging module to collect cell images and extracting contours of the collected cell images to obtain cell contour coordinates; the laser cutting module is used for emitting pulse laser, performing X-direction and Y-direction double-radio-frequency acousto-optic deflection on the pulse laser, and cutting the cell contour by using the obtained deflection laser; and the cutting control module is used for acquiring a mapping relation between the coordinate system of the microscopic imaging module and the radio frequency of the double-radio-frequency acousto-optic deflection to obtain a coordinate-frequency mapping model, and controlling the laser cutting module to perform the double-radio-frequency acousto-optic deflection on the pulse laser according to the cell contour coordinates and the coordinate-frequency mapping model. According to the invention, the cutting welting accuracy and repeatability can be improved.

Description

Technical Field

[0001] This invention belongs to the field of minimally invasive photosurgery technology and relates to an image-guided dual radiofrequency acousto-optic deflection laser cell cutting system and method. Background Technology

[0002] Cellular / subcellular scale laser microsurgery and precision cell manipulation are in high demand in single-cell omics, cell mechanobiology, neural circuit research, drug screening, and online microfluidic chip processing. Typical operations include: local cutting along cell contours, local membrane / cytoskeleton ablation, severing of intercellular connections, perforation and local permeability of subcellular regions, and microstructure writing.

[0003] To perform the above operations, three core capabilities are typically required: (1) High-resolution microscopic imaging: provides clear edge / structural information of target cells; (2) High-precision controllable energy laser actuation: achieving local effects within a safe and effective window; (3) Fast and accurate spot positioning and trajectory scanning in two-dimensional plane: the spot strictly fits the cell outline and can be repeated.

[0004] Currently, common methods for achieving minimally invasive photosurgery include galvanometer scanning, mechanical stage movement, digital microscopy / SLM projection, and manual outlining. However, these methods each have their own shortcomings and limitations, which can be summarized as follows: (1) Trajectory fitting depends on manual / semi-automatic operation, resulting in poor repeatability: cell edge extraction is unstable or requires manual correction; it is difficult to process multiple cells in batches.

[0005] (2) Scanning execution has inertia and dynamic errors: mechanical stage / galvanometer has response time, back-travel error and acceleration / deceleration limitations; microscale contour scanning is prone to corner overshoot and uneven speed, resulting in inconsistent energy deposition.

[0006] (3) Inconsistent coordinate systems lead to “seeing ≠ hitting”: The lack of strict calibration / mapping between the microscopic image coordinates and the scanning control coordinates causes the cutting trajectory to deviate from the edge. Summary of the Invention

[0007] To address the problems of laser cell cutting trajectory deviating from the cell edge in the prior art, the present invention provides an image-guided dual radio frequency acousto-optic deflection laser cell cutting system and method.

[0008] This invention is achieved through the following technical solution: In a first aspect, the present invention provides an image-guided dual-RF acousto-optic deflection laser cell cutting system, comprising: The microscopic imaging module is used to acquire cell images; The imaging control module is used to control the microscopic imaging module to acquire cell images and extract the contours of the acquired cell images to obtain cell contour coordinates. The laser cutting module is used to emit pulsed lasers and perform dual radio frequency acousto-optic deflection of the pulsed lasers in the X and Y directions, and use the deflected lasers to cut the cell contours. The cutting control module is used to obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection, and obtain the coordinate-frequency mapping model. Based on the cell contour coordinates and the coordinate-frequency mapping model, the laser cutting module is controlled to perform dual radio frequency acousto-optic deflection on the pulsed laser.

[0009] Preferably, the microscopic imaging module includes: a white light source, a first set of lenses, a dichroic mirror, a scanning objective, an imaging objective, a reflecting mirror, and a camera arranged sequentially; the light beam emitted by the white light source passes sequentially through the first set of lenses, the dichroic mirror, the scanning objective, the imaging objective, and the reflecting mirror before entering the camera; in use, the sample stage is placed between the scanning objective and the imaging objective.

[0010] Furthermore, a notch filter and a bandpass filter are provided between the reflector and the camera, and the light beam reflected by the reflector enters the camera in sequence through the notch filter and the bandpass filter.

[0011] Furthermore, the laser cutting module includes: a pulsed laser, a second set of lenses, a half-wave plate, a polarization beam splitter, and a dual radio frequency acousto-optic deflector. The pulsed laser emitted by the pulsed laser passes sequentially through the second set of lenses, the half-wave plate, the polarization beam splitter, the dual radio frequency acousto-optic deflector, the dichroic mirror, and the scanning objective lens and is incident on the sample stage.

[0012] Furthermore, the laser cutting module also includes an energy monitoring module, which includes an optical power meter and a beam splitter. The beam splitter is positioned between the dual radio frequency acousto-optic deflector and the dichroic mirror. One path of the beam splitter's outgoing light is incident on the dichroic mirror, and the other path of the outgoing light enters the optical power meter. The optical power meter is used to collect pulsed laser energy and transmit it to the cutting control module.

[0013] Furthermore, the scanning objective is an F-theta scanning objective.

[0014] Preferably, the microscopic imaging module and the laser cutting module are integrated into an optical cage system in a coaxial / quasi-coaxial manner.

[0015] In a second aspect, the present invention provides an image-guided dual-RF acousto-optic deflection laser cell cutting method, based on the system described above, comprising: Obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection to obtain the coordinate-frequency mapping model; Cell images are acquired using a microscopic imaging module, and the cell contours are extracted using an imaging control module to obtain cell contour coordinates. The laser cutting module emits pulsed laser light, and the cutting control module controls the laser cutting module to perform dual radio frequency acousto-optic deflection of the pulsed laser light in the X and Y directions according to the cell contour coordinates and coordinate-frequency mapping model. The laser cutting module then uses the deflected laser light to cut the cell contour.

[0016] Furthermore, the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection is obtained, resulting in a coordinate-frequency mapping model, specifically: The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the Y direction, and gradually adjusts the radio frequency frequency in the X direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the X-direction coordinates and the X-direction radio frequency is obtained. The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the X direction, and gradually adjusts the radio frequency frequency in the Y direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the Y-direction coordinates and the Y-direction radio frequency is obtained. Based on the correspondence between the X-axis coordinate and the X-axis radio frequency, and the correspondence between the Y-axis coordinate and the Y-axis radio frequency, a frequency-displacement model is obtained.

[0017] Furthermore, in the image-guided dual-RF acousto-optic deflection laser cell cutting method, the obtained deflection laser is divided into two paths: one path is used to cut the cell outline, and the other path is used to measure the pulse laser energy.

[0018] Compared with the prior art, the present invention has the following beneficial effects: This invention presents an image-guided dual-RF acousto-optic deflection laser cell cutting system. It establishes a mapping relationship between the coordinate system of the microscopic imaging module and the RF frequencies of the dual-RF acousto-optic deflection, providing a mathematical basis for moving the laser spot point-by-point along the contour. This ensures trajectory alignment accuracy and repeatability, improving the "cutting edge accuracy" and repeatability. The system forms a complete closed loop from cell image acquisition and cell contour extraction to frequency mapping and automatic scanning execution. This highly automated closed loop significantly reduces reliance on manual intervention.

[0019] Furthermore, the pulsed laser is split into a single pulse by a beam splitter and sent to an optical power meter to record the energy change of each pulse. This can be used to calibrate the energy window and assess long-term stability, thus making the cutting energy more controllable and safer.

[0020] Furthermore, using an F-theta scanning objective for focusing and scanning can avoid the field curvature and nonlinear displacement of traditional lenses, resulting in better scanning and focusing quality and lower distortion, making it suitable for high-precision planar scanning and cutting.

[0021] Furthermore, this invention integrates all components into a cage-like system structure, enabling precise optical axis positioning, rigid stability, and convenient disassembly and replacement. It facilitates adjustment and upgrades, and reserves space for interfaces such as fluorescence excitation / film potential, enabling secondary development. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 This is a schematic diagram of the image-guided dual-RF acousto-optic deflection laser cell cutting system in an embodiment of the present invention.

[0024] In the diagram: 1 is a white light source, 2 is the first lens, 3 is the second lens, 4 is a dichroic mirror, 5 is a scanning objective lens, 6 is a sample stage, 7 is an imaging objective lens, 8 is a notch filter, 9 is a bandpass filter, 10 is a camera, 11 is a pulsed laser, 12 is the third lens, 13 is the fourth lens, 14 is a mirror, 15 is a half-wave plate, 16 is a polarizing beam splitter, 17 is a dual RF acousto-optic deflector, 18 is a beam splitter prism, and 19 is an optical power meter. Detailed Implementation

[0025] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0026] It should be noted that the process equipment or apparatus not specifically mentioned in the following embodiments are all conventional equipment or apparatus in the art.

[0027] It should be noted that the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or apparatus that includes a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to these processes, methods, products, or apparatuses. Furthermore, unless otherwise stated, the numbering of each method step is merely a convenient tool for identifying each method step, and not intended to limit the order of the method steps or define the scope of the invention. Changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of the invention.

[0028] The image-guided dual-RF acousto-optic deflection laser cell cutting system of the present invention comprises: The microscopic imaging module is used to acquire cell images; The imaging control module is used to control the microscopic imaging module to acquire cell images and extract the contours of the acquired cell images to obtain cell contour coordinates. The laser cutting module is used to emit pulsed lasers and perform dual radio frequency acousto-optic deflection of the pulsed lasers in the X and Y directions, and use the deflected lasers to cut the cell contours. The cutting control module is used to obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection, and obtain the coordinate-frequency mapping model. Based on the cell contour coordinates and the coordinate-frequency mapping model, the laser cutting module is controlled to perform dual radio frequency acousto-optic deflection on the pulsed laser.

[0029] The microscopic imaging module of the present invention acquires cell images through wide-field illumination, objective lens imaging, and camera 10 acquisition.

[0030] In some embodiments of the present invention, the microscopic imaging module includes: a white light source 1, a first set of lenses, a dichroic mirror 4, a scanning objective 5, an imaging objective 7, a reflecting mirror 14, and a camera 10 arranged sequentially; the light beam emitted by the white light source 1 passes through the first set of lenses, the dichroic mirror 4, the scanning objective 5, the imaging objective 7, and the reflecting mirror 14 in sequence before entering the camera 10; in use, the sample stage 6 is placed between the scanning objective 5 and the imaging objective 7.

[0031] Specifically, the white light source 1 can be a DC-DC regulated fiber optic light source with continuously adjustable brightness from 0% to 100%, providing stable and controllable illumination for cell imaging. The first set of lenses includes a first lens 2 and a second lens 3. The white light output from the DC-DC regulated fiber optic light source is expanded and collimated by the two lenses to form a uniform collimated beam. After being transmitted through a dichroic mirror 4, the collimated beam is focused onto the sample surface by a scanning objective lens 5 to achieve wide-field bright-field illumination. A 10× inverted imaging objective lens 7 positioned below the sample collects the imaging light. The imaging light is refracted by a reflecting mirror 14 and finally enters the camera 10 (1-inch target surface) to acquire cell images.

[0032] In some embodiments of the present invention, a notch filter 8 and a bandpass filter 9 are further disposed between the reflector 14 and the camera 10. The light beam reflected by the reflector 14 passes sequentially through the notch filter 8 and the bandpass filter 9 before entering the camera 10. Adding the notch filter 8 and the bandpass filter 9 in front of the camera 10 can block high-energy components such as 532 nm pulsed laser light, protecting the chip of the camera 10 and improving the signal-to-noise ratio, thus providing clearer raw data for subsequent edge extraction.

[0033] In some embodiments of the present invention, the laser cutting module includes: a pulsed laser 11, a second set of lenses, a half-wave plate 15, a polarizing beam splitter (PBS) 16, and a dual radio frequency acousto-optic deflector 17. The pulsed laser emitted by the pulsed laser 11 passes sequentially through the second set of lenses, the half-wave plate 15, the PBS 16, the dual radio frequency acousto-optic deflector 17, the dichroic mirror 4, and the scanning objective lens 5 before being incident on the sample stage 6. The second set of lenses includes a third lens 12 and a fourth lens 13.

[0034] The pulsed laser 11 emits a 532 nm nanosecond pulsed laser (pulse width approximately 6 ns, repetition frequency 10 Hz). After the pulsed laser is output, it is collimated and expanded by a second set of lenses (third lens 12 and fourth lens 13). The expanded pulsed laser is polarized by a half-wave plate 15 + PBS. The polarized pulsed laser is incident on a dual radio frequency acousto-optic deflector 17 to achieve two-dimensional continuous deflection. The deflected laser is reflected by a dichroic mirror 4 and enters the scanning objective lens 5 to be focused on the target position in the sample, achieving local irradiation and cutting.

[0035] By constructing a polarization adjustment system using a half-wave plate and a PBS, the reflection / transmission beam splitting ratio can be continuously adjusted, thereby precisely controlling the pulsed laser power of the incident dual radio frequency acousto-optic deflector 17 and matching the polarization sensitivity characteristics of the dual radio frequency acousto-optic deflector 17.

[0036] This invention addresses the key challenge of "precisely controlling the focused laser beam within a two-dimensional plane and ensuring the trajectory strictly conforms to the cell contour," by constructing a scanning system consisting of dual radio frequency acousto-optic deflectors 17 and a scanning objective lens 5. To avoid significant field curvature and nonlinear displacement caused by traditional lenses, an F-theta scanning objective lens is selected to achieve high-precision, low-distortion scanning focusing within the plane (a standard scanning lens commonly used in laser marking / engraving / cutting). The deflected pulsed laser beam is focused onto the target position in the sample by the F-theta scanning objective lens, ensuring good focusing on the sample plane at different deflection angles.

[0037] The two-dimensional deflection principle of the dual radio frequency acousto-optic deflector 17 is as follows: the acousto-optic deflector changes the diffraction angle by inputting the frequency of the radio frequency signal, thereby achieving precise adjustment of the beam deflection direction and deflection angle; the dual radio frequency acousto-optic deflector 17 is driven in two orthogonal directions respectively to achieve two-dimensional plane scanning (X direction and Y direction).

[0038] In some embodiments of the present invention, the laser cutting module further includes an energy monitoring module, which includes an optical power meter 19 and a beam splitter 18. The beam splitter 18 is disposed between the dual radio frequency acousto-optic deflector 17 and the dichroic mirror 4. The emitted light from the dual radio frequency acousto-optic deflector 17 is split into two paths after passing through the beam splitter 18. One path is used to cut cells, and the other path enters the optical power meter 19. The optical power meter 19 is used to collect pulsed laser energy and transmit it to the cutting control module. The optical power meter 19 records the energy change of each pulsed laser to calibrate the cutting energy window, evaluate long-term stability, and support safe and repeatable cutting.

[0039] In some embodiments of the present invention, the microscopic imaging module and the laser cutting module are integrated into a standard 30 mm optical cage system in a coaxial / quasi-coaxial manner, realizing the coaxial / quasi-coaxial integration of the illumination, imaging, and cutting optical paths. The optical cage system can achieve precise optical axis positioning and rigid stability; it is easy to disassemble and replace; it reserves expansion interfaces and space for fluorescence excitation / film potential, which is conducive to secondary development, easy to adjust and upgrade, and the platform has strong scalability.

[0040] The image-guided dual-RF acousto-optic deflection laser cell cutting method of the present invention includes: Obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection to obtain the coordinate-frequency mapping model; Cell images are acquired using a microscopic imaging module, and the cell contours are extracted using an imaging control module to obtain cell contour coordinates. The laser cutting module emits pulsed laser light, and the cutting control module controls the laser cutting module to perform dual radio frequency acousto-optic deflection of the pulsed laser light in the X and Y directions according to the cell contour coordinates and coordinate-frequency mapping model. The laser cutting module then uses the deflected laser light to cut the cell contour.

[0041] In some embodiments of the present invention, the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection is obtained to obtain a coordinate-frequency mapping model, specifically as follows: The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the Y direction, and gradually adjusts the radio frequency frequency in the X direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the X-direction coordinates and the X-direction radio frequency is obtained. The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the X direction, and gradually adjusts the radio frequency frequency in the Y direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the Y-direction coordinates and the Y-direction radio frequency is obtained. Based on the correspondence between the X-axis coordinate and the X-axis radio frequency, and the correspondence between the Y-axis coordinate and the Y-axis radio frequency, a coordinate-frequency mapping model is obtained.

[0042] In some embodiments of the present invention, the obtained deflected laser is split into two paths: one path is used to cut the cell outline, and the other path is used to measure the pulsed laser energy.

[0043] The imaging control module of this invention controls image acquisition and contour extraction to obtain cell contour coordinates, and the cutting control module is used for coordinate mapping, cutting trajectory planning, radio frequency output and status monitoring to form a closed-loop process.

[0044] The imaging control module adopts a modular design, with an interface including a camera control area, parameter setting area, image display area, and status display area. It supports online adjustment of exposure / gain / frame rate, filter threshold, etc., and provides error prompts. The image processing flow is as follows: image acquisition → color space conversion → grayscale conversion → contrast enhancement → noise suppression → cell edge detection → morphological processing → cell contour extraction → contour coordinates. It achieves a processing speed of approximately 15 frames per second through a multi-threaded mechanism, supporting real-time interaction and rapid iteration to meet real-time requirements. It also supports image saving, online parameter adjustment, and result export. Cell edge detection uses methods such as Canny. The output format of the cell contour is a set of coordinate points (pixels or physical units), which serves as the input to the cutting control module.

[0045] To ensure that the "seen position" is consistent with the "hit position", this invention performs coordinate-frequency calibration, and on this basis, establishes a one-to-one correspondence between the camera coordinate system and the dual radio frequency of the dual radio frequency acousto-optic deflector, and obtains the coordinate-frequency mapping model through linear fitting.

[0046] The goal of coordinate-frequency calibration is to establish a quantitative correspondence between any point (X, Y) in the camera coordinate system and the combination of radio frequency (f1, f2) of the dual radio frequency acousto-optic deflectors, ensuring that each image coordinate can be uniquely and stably converted into executable radio frequency parameters.

[0047] Coordinate-frequency calibration process: This involves synchronously acquiring the corresponding data of "laser spot displacement in camera 10" and "dual radio frequency of dual radio frequency acousto-optic deflectors 17," and then performing linear fitting and calibration. Specifically, the method is as follows: The radio frequency in the Y direction is fixed, and the radio frequency in the X direction is scanned to obtain the spot displacement curve in camera 10; then, the channels are switched to repeatedly acquire the coordinate-frequency mapping relationship in the Y direction; after forming the coordinate-frequency mapping model, it can be used for automatic coordinate-to-radio frequency mapping and execution. This standardized and reusable calibration process based on coordinate-frequency calibration supports migration applications across different platforms and AOD devices.

[0048] After importing the cell contour coordinates, the cutting control module automatically maps the coordinates to the radio frequency and plans the cutting trajectory, controlling the dual radio frequency acousto-optic deflectors 17 to perform radio frequency acousto-optic deflection according to the cutting trajectory.

[0049] The cutting control module supports importing cell contour point sets from the microscopic imaging module; it can import and manage no fewer than 20 sets of cell contours at a time, making it suitable for batch cutting.

[0050] The automatic coordinate-to-RF frequency mapping works by automatically converting coordinates (X,Y) into frequency sequences (f1,f2) based on the calibrated coordinate-frequency mapping model. This process is transparent to the user, reduces operational complexity, and avoids human calculation errors.

[0051] Laser spot motion path planning and execution: Each coordinate point is visited in the order of "X direction first, then Y direction". By setting the radio frequency step size and dwell time, two scanning cutting modes, namely point-by-point or continuous, are realized along the cell edge to complete precise cutting.

[0052] The cutting control module displays the radio frequency of the dual radio frequency acousto-optic deflectors 17, the target coordinate point number, and the system status in real time, and provides anomaly alerts to assist in troubleshooting.

[0053] The imaging control module and the cutting control module work together to achieve complete closed-loop control from "image acquisition and contour extraction" to "mapping and automatic spot scanning".

[0054] Example The image-guided dual-RF acousto-optic deflection laser cell cutting system of this embodiment includes: a microscopic imaging module, an imaging control module, a laser cutting module, and a cutting control module.

[0055] The microscopic imaging module includes, in sequence: a white light source 1, a first lens 2, a second lens 3, a dichroic mirror 4, a scanning objective 5, an imaging objective 7, a reflecting mirror 14, a notch filter 8, a bandpass filter 9, and a camera 10. The light beam emitted from the white light source 1 passes sequentially through the first lens 2, the second lens 3, the dichroic mirror 4, the scanning objective 5, the imaging objective 7, the reflecting mirror 14, the notch filter 8, and the bandpass filter 9 before entering the camera 10. In use, the sample stage 6 is placed between the scanning objective 5 and the imaging objective 7. The scanning objective 5 is an F-theta scanning objective.

[0056] The laser cutting module includes: a pulsed laser 11, a third lens 12, a fourth lens 13, a half-wave plate 15, a PBS 16, a dual radio frequency acousto-optic deflector 17, an optical power meter 19, and a beam splitter 18. The pulsed laser emitted by the pulsed laser 11 passes sequentially through the third lens 12, the fourth lens 13, the half-wave plate 15, and the PBS before entering the dual radio frequency acousto-optic deflector 17. The outgoing light from the dual radio frequency acousto-optic deflector 17 is split into two paths after passing through the beam splitter 18. One path passes through the dichroic mirror 4 and the scanning objective lens 5 and is incident on the sample stage 6, while the other path enters the optical power meter 19.

[0057] The specific working method of the image-guided dual-RF acousto-optic deflection laser cell cutting system in this embodiment is as follows: (1) Construction of coordinate-frequency mapping model The pulsed laser 11 is turned on to emit pulsed laser light. The Y-direction radio frequency of the dual radio frequency acousto-optic deflector 17 is fixed and the X-direction radio frequency is continuously changed by the cutting control module. The pulsed laser light emitted by the pulsed laser passes sequentially through the third lens 12, the fourth lens 13, the half-wave plate 15, the PBS 16, the dual radio frequency acousto-optic deflector 17, the dichroic mirror 4, the scanning objective lens 5, the imaging objective lens 7, the reflecting mirror 14, the notch filter 8, and the bandpass filter 9 before entering the camera 10, thus obtaining the displacement curve of the laser spot in the X-direction. Similarly, the X-direction radio frequency of the dual radio frequency acousto-optic deflector 17 is fixed and the Y-direction radio frequency is continuously changed by the cutting control module, thus obtaining the displacement curve of the laser spot in the Y-direction, forming a coordinate-frequency mapping model.

[0058] (2) Obtaining cell contour coordinates The pulsed laser 11 is turned off, and the white light source 1 is turned on. The beam emitted by the white light source 1 passes sequentially through the first lens 2, the second lens 3, the dichroic mirror 4, the scanning objective lens 5, the imaging objective lens 7, the reflecting mirror 14, the notch filter 8, and the bandpass filter 9 before entering the camera 10. The cell image acquired by the camera 10 is transmitted to the imaging control module. The imaging control module performs color space conversion, grayscale conversion, contrast enhancement, noise suppression, cell edge detection, morphological processing, and cell contour extraction on the cell image in sequence to obtain the cell contour coordinates and transmit them to the cutting control module.

[0059] (3) Cell cutting The cutting control module performs coordinate-frequency mapping based on cell contour coordinates and a coordinate-frequency mapping model, thereby planning the cutting trajectory and the corresponding radio frequency.

[0060] The white light source 1 and camera 10 are turned off, and the pulsed laser 11 is turned on. The cutting control module controls the dual radio frequency acousto-optic deflector 17 to acousto-optic deflect the pulsed laser according to the cutting trajectory and the corresponding radio frequency. The deflected pulsed laser dichroic mirror 4 and scanning objective 5 irradiate the sample, thereby cutting the cells in the sample.

[0061] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of this invention.

Claims

1. An image-guided dual-RF acousto-optic deflection laser cell cutting system, characterized in that, include: The microscopic imaging module is used to acquire cell images; The imaging control module is used to control the microscopic imaging module to acquire cell images and extract the contours of the acquired cell images to obtain cell contour coordinates. The laser cutting module is used to emit pulsed lasers and perform dual radio frequency acousto-optic deflection of the pulsed lasers in the X and Y directions, and use the deflected lasers to cut the cell contours. The cutting control module is used to obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection, and obtain the coordinate-frequency mapping model. Based on the cell contour coordinates and the coordinate-frequency mapping model, the laser cutting module is controlled to perform dual radio frequency acousto-optic deflection on the pulsed laser.

2. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 1, characterized in that, The microscopic imaging module includes: a white light source (1), a first set of lenses, a dichroic mirror (4), a scanning objective (5), an imaging objective (7), a reflecting mirror (14), and a camera (10) arranged in sequence; the light beam emitted by the white light source (1) passes through the first set of lenses, the dichroic mirror (4), the scanning objective (5), the imaging objective (7), and the reflecting mirror (14) in sequence before entering the camera (10); in use, the sample stage (6) is placed between the scanning objective (5) and the imaging objective (7).

3. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 2, characterized in that, A notch filter (8) and a bandpass filter (9) are also provided between the reflector (14) and the camera (10). The light beam reflected by the reflector (14) enters the camera (10) in sequence through the notch filter (8) and the bandpass filter (9).

4. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 2, characterized in that, The laser cutting module includes: a pulsed laser (11), a second set of lenses, a half-wave plate (15), a polarization beam splitter (16), and a dual radio frequency acousto-optic deflector (17). The pulsed laser emitted by the pulsed laser (11) is incident on the sample stage (6) through the second set of lenses, the half-wave plate (15), the polarization beam splitter (16), the dual radio frequency acousto-optic deflector (17), the dichroic mirror (4), and the scanning objective (5).

5. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 4, characterized in that, The laser cutting module also includes an energy monitoring module, which includes an optical power meter (19) and a beam splitter (18). The beam splitter (18) is positioned between the dual radio frequency acousto-optic deflector (17) and the dichroic mirror (4). One output light from the beam splitter (18) is incident on the dichroic mirror (4), and the other output light enters the optical power meter (19). The optical power meter (19) is used to collect pulsed laser energy and transmit it to the cutting control module.

6. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 2, characterized in that, The scanning objective (5) is an F-theta scanning objective.

7. The image-guided dual-RF acousto-optic deflection laser cell cutting system according to claim 1, characterized in that, The microscopic imaging module and the laser cutting module are integrated into an optical cage system in a coaxial / quasi-coaxial manner.

8. An image-guided dual-RF acousto-optic deflection laser cell cutting method, characterized in that, The system based on any one of claims 1-7 comprises: Obtain the mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection to obtain the coordinate-frequency mapping model; Cell images are acquired using a microscopic imaging module, and the cell contours are extracted using an imaging control module to obtain cell contour coordinates. The laser cutting module emits pulsed laser light, and the cutting control module controls the laser cutting module to perform dual radio frequency acousto-optic deflection of the pulsed laser light in the X and Y directions according to the cell contour coordinates and coordinate-frequency mapping model. The laser cutting module then uses the deflected laser light to cut the cell contour.

9. The image-guided dual-RF acousto-optic deflection laser cell cutting method according to claim 8, characterized in that, The mapping relationship between the coordinate system of the microscopic imaging module and the radio frequency of the dual radio frequency acousto-optic deflection is obtained, resulting in a coordinate-frequency mapping model, specifically: The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the Y direction, and gradually adjusts the radio frequency frequency in the X direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the X-direction coordinates and the X-direction radio frequency is obtained. The laser cutting module emits pulsed lasers, fixes the radio frequency frequency in the X direction, and gradually adjusts the radio frequency frequency in the Y direction to perform dual radio frequency acousto-optic deflection on the pulsed lasers. The coordinates of the deflected pulsed laser spot are obtained using a microscopic imaging module, and the correspondence between the Y-direction coordinates and the Y-direction radio frequency is obtained. Based on the correspondence between the X-axis coordinate and the X-axis radio frequency, and the correspondence between the Y-axis coordinate and the Y-axis radio frequency, a frequency-displacement model is obtained.

10. The image-guided dual-RF acousto-optic deflection laser cell cutting method according to claim 8, characterized in that, The resulting deflected laser is split into two paths: one path is used to cut the cell outline, and the other path is used to measure the pulsed laser energy.

Images