CN116519601A - Photoacoustic microscopic imaging system and method based on Airy light beam combined sparse sampling - Google Patents
Photoacoustic microscopic imaging system and method based on Airy light beam combined sparse sampling Download PDFInfo
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Abstract
The invention discloses a photoacoustic microscopic imaging system and a photoacoustic microscopic imaging method based on Airy light beam combined sparse sampling, which are suitable for scenes with large depth of field and rapid imaging monitoring, wherein the system comprises: the beam generation module is used for generating nanosecond pulse Gaussian beams and processing the nanosecond pulse Gaussian beams to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam; the beam shaping module is used for carrying out polarization adjustment, phase modulation, filtering, focusing and Fourier transform on the first standard linear polarization Gaussian beam to obtain an Airy beam; the system control and sparse scanning module is used for sampling the two-dimensional sparse scanning of the object excited with the Airy light beam by using a two-dimensional triangular lissajous track when receiving the second standard linear polarized Gaussian light beam, so as to obtain a sparse photoacoustic signal; the signal receiving and processing module is used for carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image; the image reconstruction module is used for reconstructing the sparse image by adopting a super-resolution reconstruction network to obtain a high-resolution photoacoustic microscopic original image.
Description
Technical Field
The invention belongs to the technical field of optical microscopic imaging, and particularly relates to a photoacoustic microscopic imaging system and method based on Airy light beam combined sparse sampling.
Background
Optical resolution photoacoustic microscopy (OR-PAM) is an emerging biomedical imaging technology that has evolved dramatically in recent years to enable label-free in vivo dissection and functional imaging of microvasculature, lipids and melanin in biological tissues. OR-PAM has an inherent depth resolution capability that benefits from time-of-flight information captured by each photoacoustic signal excited by an a-scan, which allows volumetric imaging using only two-dimensional (2D) raster scanning. In a typical OR-PAM system, a highly focused gaussian beam is selected using an optical objective lens for lateral resolution of the optical diffraction limit. However, due to diffraction limits, the conventionally employed gaussian beam diverges rapidly along its propagation direction, resulting in a very short depth of focus for photoacoustic signal excitation. The limited depth of focus makes it impossible to acquire volumetric images with consistent lateral resolution in the depth direction to accurately acquire the structural, functional and metabolic status of the biological system.
A beam field that is constant in propagation (e.g., a bessel beam) can expand the depth of focus without affecting the focus size. However, bessel beam-based methods typically obtain poor image contrast because the lateral annular sidelobe structures can create background artifacts and reduce axial resolution. Furthermore, the bessel beam has only a small fraction of the light energy (< 20%) in its central main lobe, which affects the sensitivity and image quality of the acquired signal. Furthermore, rapid screening and reproducible monitoring are necessary requirements for clinical diagnosis of many diseases, whereas full sampling imaging based on conventional raster scanning is slow, especially without sacrificing spatial resolution and with large field of view imaging requirements.
Disclosure of Invention
In order to solve the problems in the related art, the invention provides a photoacoustic microscopy imaging system and a photoacoustic microscopy imaging method based on Airy light beam combined sparse sampling. The technical problems to be solved by the invention are realized by the following technical scheme:
the invention provides a photoacoustic microscopic imaging system based on Airy light beam joint sparse sampling, which comprises:
the light beam generation module is used for generating a nanosecond pulse type Gaussian beam, and processing the nanosecond pulse type Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam;
the beam shaping module is used for carrying out polarization adjustment, phase modulation, filtering, focusing and Fourier transformation on the first standard linear polarization Gaussian beam to obtain an Airy beam excited to an object;
the system control and sparse scanning module is used for carrying out two-dimensional sparse scanning sampling on the object excited with the Airy light beam by using a two-dimensional triangular lissajous track when the second standard linear polarized Gaussian light beam is received, so as to obtain a sparse photoacoustic signal;
the signal receiving and processing module is used for receiving the sparse photoacoustic signal and carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image;
the image reconstruction module is used for carrying out image reconstruction on the sparse image by adopting a pre-trained super-resolution reconstruction network to obtain a high-resolution photoacoustic microscopic original image of the object; wherein the super-resolution reconstruction network comprises: two discrimination networks, an up-sampling network and a down-sampling network; the up-sampling network and one discrimination network form a forward GAN network, and the down-sampling network and the other discrimination network form a reverse GAN network.
In some embodiments, the upsampling network sequentially comprises: a first convolution layer, an activation layer, a residual sub-network, a sub-pixel convolution layer, and a second convolution layer; the downsampling network comprises, in order: the first convolution layer, the active layer, the residual sub-network, a first downsampled convolution layer, a second downsampled convolution layer, and the second convolution layer; the two discrimination networks are discrimination networks in the super-resolution generation countermeasure network.
In some embodiments, the system control and sparse scanning module comprises: the system comprises a photoelectric detector, a scanning galvanometer, a data acquisition card and a system control module;
the photoelectric detector is used for synchronously sending a trigger signal to the scanning galvanometer and the data acquisition card when receiving the second standard linear polarization Gaussian beam;
the data acquisition card and the system control module are used for generating two-dimensional triangular waveforms at preset frequency and respectively sending the two triangular waveforms to the two vibrating mirrors of the scanning vibrating mirror when the trigger signal is received;
the scanning galvanometer is used for carrying out two-dimensional sparse scanning sampling on the object according to the two-dimensional triangular waveforms received by the two galvanometers and the two-dimensional triangular lissajous tracks to obtain the sparse photoacoustic signal.
In some embodiments, the beam generation module comprises, in order:
a nanosecond pulse laser for generating a nanosecond pulse gaussian beam;
the polarizer is used for carrying out polarization adjustment on the nanosecond pulse Gaussian beam to obtain a standard linear polarization Gaussian beam;
a polarizing beam splitter for dividing the standard linearly polarized gaussian beam into a first standard linearly polarized gaussian beam and a second standard linearly polarized gaussian beam according to a preset ratio, and taking the second standard linearly polarized gaussian beam as a second standard linearly polarized gaussian beam;
the diaphragm is used for optimizing the first standard linear polarization Gaussian beam to obtain an optimized first standard linear polarization Gaussian beam;
and the beam expanding lens group is used for expanding the optimized first standard linear polarization Gaussian beam to obtain the first standard linear polarization Gaussian beam.
In some embodiments, the beam expanding lens group comprises, in order: the first converging lens, the first reflecting mirror, the first small hole and the second converging lens;
the first small hole is used for filtering other stray light in the standard linear polarization Gaussian beam passing through the first converging lens.
In some embodiments, the beam shaping module comprises, in order: the device comprises a half-wave plate, a spatial light modulator, a beam shrinking lens group, a scanning array mirror, an objective lens and a three-dimensional moving platform;
the half-wave plate is used for adjusting the polarization direction of the first standard linear polarization Gaussian beam;
the spatial light modulator is used for carrying out phase modulation on the polarized first standard linear polarization Gaussian beam according to preset cube phase modulation parameters to obtain a modulated beam;
the beam shrinking lens group is used for carrying out beam shrinking and filtering treatment on the modulated light beam;
the objective lens is used for focusing and Fourier transforming the light beam passing through the scanning array mirror to obtain the Airy light beam;
the three-dimensional moving platform is used for placing the object and controlling the object to be positioned at the focus of the objective lens through the movement of the three-dimensional moving platform in the horizontal and/or vertical directions.
In some embodiments, the spatial light modulator is loaded with phase blazed gratings of different grating periods, and is further configured to diffract the modulated light beam to different diffraction angles to separate the zero-order light spot and the first-order light spot;
the beam shrinking lens group sequentially comprises: a third converging lens, a second aperture and a fourth converging lens;
and the second small hole is used for filtering the zero-order light spot.
In some embodiments, the beam shaping module further comprises: a third mirror and a fourth mirror; the third reflecting mirror and the fourth reflecting mirror are sequentially positioned between the beam shrinking lens group and the scanning array mirror;
the third reflector is used for reflecting the light beam passing through the beam shrinking lens group to the fourth reflector;
the fourth reflector is used for reflecting the light beam reflected by the third reflector to the scanning array mirror.
In some embodiments, the signal receiving and processing module comprises:
the ultrasonic transducer is used for receiving the sparse photoacoustic signal and obtaining an electric signal according to the sparse photoacoustic signal;
and the amplifier is used for amplifying the electric signal to obtain the amplified sparse photoacoustic signal.
The invention also provides a photoacoustic microscopic imaging method based on Airy light beam combined sparse sampling, which is applied to the photoacoustic microscopic imaging system and comprises the following steps:
generating a nanosecond pulse Gaussian beam, and processing the nanosecond pulse Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam;
performing polarization adjustment, phase modulation, filtering, focusing and Fourier transformation on the first standard linear polarization Gaussian beam to obtain an Airy beam excited to an object;
when the Gaussian beam is polarized according to the second standard line, carrying out two-dimensional sparse scanning sampling on the object excited with the Airy beam by using a two-dimensional triangular Lissajous track to obtain a sparse photoacoustic signal;
receiving the sparse photoacoustic signal, and carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image;
and adopting a pre-trained super-resolution reconstruction network to reconstruct the image of the sparse image to obtain a high-resolution photoacoustic microscopic original image of the object.
The invention has the following beneficial technical effects:
aiming at the problem of mutual restriction among spatial resolution, depth of field, detection sensitivity and imaging speed in the current photoacoustic microscopy imaging, on one hand, the diffraction-free Airy light beam obtained by the light beam generating module and the light beam shaping module has the characteristics of longer focal depth and better focusing effect, and the main lobe of the diffraction-free Airy light beam can carry relatively more energy (about 50%), so that the range of axial imaging of an object (such as biological tissue) can be remarkably expanded, thereby realizing large depth of field and high-sensitivity photoacoustic microscopy imaging, and obtaining a high-resolution imaging image with unchanged depth-direction resolution; on the other hand, the invention carries out two-dimensional sparse scanning sampling on the object excited with the Airy light beam by using a two-dimensional triangular Lissajous track, and carries out image reconstruction by the pre-trained super-resolution reconstruction network provided by the invention after processing the scanning sampling signal, thereby meeting the imaging requirements of high-efficiency and repeatable monitoring. Therefore, the invention can realize the rapid imaging monitoring with large depth of field and high sensitivity on objects (such as physiological and pathological activities).
The present invention will be described in further detail with reference to the accompanying drawings and examples.
Drawings
Fig. 1 is a schematic structural diagram of a photoacoustic microscopic imaging system based on Airy beam joint sparse sampling according to an embodiment of the present invention;
fig. 2 is another schematic structural diagram of an exemplary optical acoustic microscopic imaging system based on Airy beam joint sparse sampling according to an embodiment of the present invention;
FIG. 3A is a diagram illustrating a comparison of an exemplary conventional point-by-point raster scan design provided by an embodiment of the present invention with a schematic diagram of the structure of the sparse scan design employed by the present invention;
FIG. 3B is a schematic waveform diagram of an exemplary triangular wave provided by an embodiment of the present invention;
FIG. 4A is a block diagram illustrating an exemplary SR-cycleGAN network provided by embodiments of the present invention;
fig. 4B is a schematic structural diagram of an exemplary g_down network according to an embodiment of the present invention;
fig. 4C is a schematic structural diagram of an exemplary residual block according to an embodiment of the present invention;
fig. 5 is a schematic diagram of effects related to an exemplary optical acoustic microscopic imaging system based on Airy beam joint sparse sampling according to an embodiment of the present invention;
fig. 6 is a flowchart of a photoacoustic microimaging method based on Airy beam joint sparse sampling according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but embodiments of the present invention are not limited thereto.
In the description of the present invention, the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Further, one skilled in the art can engage and combine the different embodiments or examples described in this specification.
Although the invention is described herein in connection with various embodiments, other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed invention, from a study of the drawings, the disclosure, and the appended claims. In the claims, the word "comprising" does not exclude other elements or steps, and the "a" or "an" does not exclude a plurality. A single processor or other unit may fulfill the functions of several items recited in the claims. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.
Fig. 1 is a schematic structural diagram of a photoacoustic microimaging system based on Airy beam joint sparse sampling according to an embodiment of the present invention, as shown in fig. 1, the photoacoustic microimaging system includes: the system comprises a light beam generating module 1, a system control and sparse scanning module 2, a signal receiving and processing module 3, a signal receiving and processing module 4 and an image reconstruction module 5.
The beam generation module 1 is used for generating a nanosecond pulse type Gaussian beam, and processing the nanosecond pulse type Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam. And the beam shaping module 2 is used for carrying out polarization adjustment, phase modulation, filtering, focusing and Fourier transformation on the first standard linear polarization Gaussian beam to obtain the Airy beam excited to the object. And the system control and sparse scanning module 3 is used for carrying out two-dimensional sparse scanning sampling on the object excited with the Airy light beam by using a two-dimensional triangular lissajous track when receiving the second standard linear polarization Gaussian light beam so as to obtain a sparse photoacoustic signal. And the signal receiving and processing module 4 is used for receiving the sparse photoacoustic signal and carrying out maximum value projection according to the sparse photoacoustic signal to obtain a sparse image. The image reconstruction module 5 is used for performing image reconstruction on the sparse image by adopting a pre-trained super-resolution reconstruction network to obtain a high-resolution photoacoustic microscopic original image of the object; wherein, super resolution rebuilding network includes: two discrimination networks, an up-sampling network and a down-sampling network; the up-sampling network and one discrimination network form a forward GAN network, and the down-sampling network and the other discrimination network form a reverse GAN network.
In some embodiments, as shown in fig. 2, the beam generation module sequentially comprises: a nanosecond pulse laser 11 for generating a nanosecond pulse gaussian beam; a polarizer 12 for performing polarization adjustment on the nanosecond pulse gaussian beam to obtain a standard linear polarized gaussian beam; a polarizing beam splitter 13 for splitting the standard linear polarized gaussian beam into a first standard linear polarized gaussian beam (90% beam energy) and a second standard linear polarized gaussian beam (10% beam energy) according to a preset ratio (e.g., 9:1), and taking the second standard linear polarized gaussian beam as the second standard linear polarized gaussian beam for triggering the system control and sparse scanning module 2. A diaphragm 14, configured to optimize the first standard linearly polarized gaussian beam, to obtain an optimized first standard linearly polarized gaussian beam; and the beam expansion lens group 15 is used for expanding the optimized first standard linear polarization Gaussian beam to obtain the first standard linear polarization Gaussian beam. Specifically, as shown in fig. 2, the beam expander lens group 15 sequentially includes: the first converging lens 151, the first reflecting mirror 152, the first small hole 153 (not shown in fig. 2) and the second converging lens 154, wherein the first converging lens 151 and the second converging lens 154 are respectively used for focusing or diverging light beams, the light spot diameter (simply called beam expansion) can be enlarged through the combined use of the two converging lenses, and the first small hole 153 is used for filtering other stray light in the standard linear polarized Gaussian light beams passing through the first converging lens 151 so as to finally improve the signal-to-noise ratio of the photoacoustic signals, thereby further enhancing the definition of the finally obtained high-resolution photoacoustic microscopic original image. In some embodiments, as shown in fig. 2, a neutral density filter 16 is further included between the nanosecond pulse laser 11 and the polarizer 12, where the neutral density filter 16 is used to perform energy adjustment on the nanosecond pulse gaussian beam generated by the nanosecond pulse laser 11, and the nanosecond pulse gaussian beam after energy adjustment is transmitted to the polarizer 12, and is subjected to polarization adjustment by the polarizer 12.
Exemplary nanosecond pulsed excitation light wavelengths are 532nm with repetition frequencies of 1KHz-50KHz. The repetition frequency of the nanosecond pulse type excitation light can be adjusted according to actual use conditions.
In some embodiments, as shown in fig. 2, the beam shaping module 2 comprises, in order: a half wave plate 21, a spatial light modulator 22, a beam shrinking lens group 23, a scanning matrix mirror 24, an objective lens 25 and a three-dimensional moving platform 26 (not shown in fig. 2). A half-wave plate 21 for adjusting the polarization direction of the first standard linearly polarized gaussian beam; a spatial light modulator 22, configured to perform phase modulation on the modulated first standard linear polarization gaussian beam according to a preset cubic phase modulation parameter, to obtain a modulated beam; specifically, the spatial light modulator 22 is further loaded with phase blazed gratings with different grating periods on the basis of the cubic phase diagram, and is used for not only obtaining a modulated light beam, but also diffracting the modulated light beam to different diffraction angles, so as to separate a zero-order light spot and a first-order light spot. A beam reduction lens group 23 for performing beam reduction and filtering processing on the light beam passing through the spatial light modulator 22; specifically, as shown in fig. 2, the beam reduction lens group 23 sequentially includes: the third converging lens 231, the second small hole 232 and the fourth converging lens 233, wherein the third converging lens 231 and the fourth converging lens 233 can reduce the diameter of the light spot when combined, and the second small hole 232 is used for filtering out the zero-order light spot. The light beam passing through the beam shrinking lens group 23 can reach the scanning array mirror 24, the light beam passing through the scanning array mirror 24 reaches the objective lens 25, and the objective lens 25 is used for focusing and Fourier transforming the light beam passing through the scanning array mirror 24 to obtain the Airy light beam. The three-dimensional moving platform 26 located at the rear end of the objective lens 25 is used for placing the object 6, and controls the object 6 to be located at the focus of the objective lens 25 through the movement of the three-dimensional moving platform 26 in the horizontal and/or vertical directions, that is, the object can be driven to move in two dimensions in the horizontal direction to adjust the excitation light to focus on and irradiate the region of interest on the object 6, and the object can be driven to move vertically up and down in the Z-axis direction through the three-dimensional moving platform 26, so that the focus of the excitation light is accurately irradiated on the target site of the object 6. In some embodiments, as shown in fig. 2, after the half-wave plate 21, a second mirror 27 is further included for reflecting the light beam passing through the half-wave plate 21 to the spatial light modulator 22. In some embodiments, as shown in fig. 2, there are a third mirror 28 and a fourth mirror 28 between the fourth condensing lens 233 and the scanning array mirror 24, the third mirror 28 being configured to reflect the light beam passing through the fourth condensing lens 233 to a fourth mirror 29, and the fourth mirror 29 being configured to reflect the light beam reflected by the third mirror 28 to the scanning array mirror 24.
Here, the above-mentioned cube phase modulation parameters (e.g., fringe spacing, modulation depth, etc.) of the spatial light modulator 22 may have multiple groups for modulating light beams having different focal lengths, main lobe sizes, and focused light energy, respectively.
Here, the numerical aperture parameters of the objective lens 25 may be determined according to the object requirements to be observed, thereby controlling the image reconstruction module 5 to perform trans-scale high resolution imaging of the object of interest at different scales. Illustratively, the Numerical Aperture (NA) of the objective lens 25 may be 0.1 and the multiple may be 4.
In some embodiments, as shown in fig. 2, the system control and sparse scanning module 3 comprises: the photoelectric detector 31, the scanning galvanometer 24, the data acquisition card 32 and the system control module 33, and the photoelectric detector 31 is electrically connected with the data acquisition card 32. The photodetector 31 is used for sending a trigger signal to the scanning galvanometer 24 and the data acquisition card 32 at the same time when receiving the second standard linear polarization Gaussian beam; the system control module 33 is configured to generate two-dimensional triangular waveforms at a preset frequency and send the two-dimensional triangular waveforms to two vibrating mirrors of the scanning vibrating mirror 24 respectively when the data acquisition card 32 receives the trigger signal, where the scanning vibrating mirror 24 is configured to sample the object 6 by two-dimensional sparse scanning according to the two-dimensional triangular waveforms received by the two vibrating mirrors and by two-dimensional triangular lissajous tracks, so as to obtain sparse photoacoustic signals.
Here, the image reconstruction process includes: (a) Photoacoustic waves generated from an absorber (e.g., biological tissue) at each position in the depth direction propagate in sequence to reach the absorber surface and are received and collected by a signal receiving means; (b) Converting the flight time into depth information by measuring the intensity change of the photoacoustic signal corresponding to the flight time of the acoustic wave, and then forming a one-dimensional depth resolution image of 'A-scan'; (c) Then, respectively reconstructing two-dimensional or three-dimensional tomographic images by two-dimensional scanning on the surface of the absorber; (d) The x-y plane photoacoustic microscopy image can be reconstructed from the maximum projection of the raw 3D data. To ensure high spatial resolution image quality, the full sampling scan step in photoacoustic imaging needs to be less than half the lateral resolution of the system according to the nyquist sampling theorem. Taking imaging a tumor as an example, the tumor region of interest is typically about 4mm by 4mm, the scan point is 668 by 668, the scan step distance is 6 μm, and a full sample dataset is to be acquired from 300 sets of microscopic images according to this standard, as shown in fig. 3A (a). The acquisition of sparse sample data corresponding to full sample data is intended to select a two-dimensional triangular lissajous trajectory as the optimal sparse sample pattern, and such a scan design may provide a more uniform sample density than a sinusoidal lissajous trajectory, as shown in fig. 3A (b). This sparse sampling approach requires three axes to follow the triangular waveform while precisely controlling the frequency. The waveforms on the spatial axes (X, Y) may be laser positioned by sending triangular waveforms to a pair of galvanometers (GM 1, GM 2). In the scanning, two waveforms can be generated by using the multifunctional data acquisition card at a specific selected frequency and sent to the two vibrating mirrors, so that sparse scanning data are generated, as shown in fig. 3B, which is a waveform of a triangular wave output by the data acquisition card on a scanning axis.
In some embodiments, as shown in fig. 2, the signal receiving and processing module 4 includes: the ultrasonic transducer 41 is located between the object 6 and the objective lens 25, and is configured to receive the sparse photoacoustic signal, and obtain an electrical signal from the sparse photoacoustic signal. The ultrasonic transducer is exemplified by a center frequency of 50MHz and a bandwidth of 78%. The amplifier 42 is electrically connected to the ultrasonic transducer 41 and the data acquisition card 32, and is configured to amplify the electrical signal sent by the ultrasonic transducer 41, obtain an amplified sparse photoacoustic signal, and send the sparse photoacoustic signal to the data acquisition card 32. And carrying out maximum projection according to the amplified and collected sparse photoacoustic signals to obtain sparse images, wherein the amplified signals can be more than 50 mv.
In some embodiments, the image reconstruction module 5 includes an imaging terminal 51, where the imaging terminal 51 is also electrically connected to the data acquisition card 32, and is configured to receive sparse signal data transmitted by the data acquisition card, and perform image reconstruction on a sparse image by using a pre-trained super-resolution reconstruction network, so as to obtain a high-resolution photoacoustic microscopic original image of the object. The system control module 33 is included in the imaging terminal 51.
In some embodiments, imaging terminal 51 is also configured to send control signals to nanosecond pulsed laser 11 to control the switching on of nanosecond pulsed laser 11.
Here, the network structure of the pre-trained super-resolution reconstruction network (SR-cycleGAN network) is shown in fig. 4A, in which an up-sampling network (g_up) and a discrimination network Dx constitute a forward GAN network, a down-sampling network (g_down) and a discrimination network Dy constitute a reverse GAN network, and the g_up network and the g_down network are used to generate a high-resolution image and a low-resolution image, respectively. The discrimination network Dx and the discrimination network Dy are two discrimination networks in the SRGAN network.
Specifically, the g_up network sequentially includes: a first convolution layer, an activation layer, a residual sub-network, a sub-pixel convolution layer, and a second convolution layer. The g_down network comprises in order: the system comprises a first convolution layer, an activation layer, a residual sub-network, a first downsampling convolution layer, a second downsampling convolution layer and a second convolution layer; the two discrimination networks are discrimination networks in the super-resolution generation countermeasure network. Illustratively, as shown in fig. 4B, the first convolution layer has a parameter k9n64s1, which represents a convolution kernel 9*9, a dimension 64, a step size 1, an active layer a pralu layer, and the residual sub-network may include 16 residual blocks, each of which has a structure as shown in fig. 4C. The parameters of the first downsampling convolution layer and the second downsampling convolution layer are k2n64s2, and the parameters of the second convolution layer are k2n3s1. The G_up network is an SRResNet network, and the sub-pixel convolution layer in the G_up is replaced by a first downsampling convolution layer and a second downsampling convolution layer, so that the G_down structure can be obtained.
Specifically, the forward GAN network implements mapping of the X image domain to the Y image domain, and the reverse GAN network implements mapping of the Y image domain to the X image domain. Since the two are in inverse mapping relation, x→g_up (x) →g_down (g_up (x))x can be realized by forward cyclic consistency loss, and y→g_down (y) →g_up (g_down (y))y can be realized by reverse consistency loss. Based on the above analysis, the countermeasures loss can be applied to the two mapping functions, for the mapping function G_up: X→Y and the discrimination network D Y The objective function can be expressed as formula (1):
D Y (y) represents the discrimination network D Y The discrimination value for the true sample image y,represents the expected value, D Y (G_up (x)) represents the discrimination network D Y For the discrimination value of the generated image g_up (x), g_up (x) is the image generated by the g_up network. The goal of the G_up network is to attempt to generate an image G_up (x) that is similar to the Y domain image, while D Y The purpose of (c) is to distinguish between g_up (x) and real sample y. For mapping function G_Down Y→X and its discrimination network D X Also introduces similar countermeasures, i.e. L GAN (G_down,D X X, Y). Since it is difficult to guarantee that a single input can output the desired result with a simple counterloss constraint, a cyclic consistency loss is added to ensure the cyclic consistency of the mapping relationship, namely the following formula (2):
g_down (y) is an image generated by the g_down network, g_up (g_down (y)) represents an image generated by the g_up network from the input image g_down (y), and g_down (g_up (x)) represents an image generated by the g_down network from the input image g_up (x). The objective function of the whole network is thus equation (3):
wherein L is GAN (G_up,D Y X, Y) is the objective function of the forward GAN network, L GAN (G_down,D X X, Y) is an objective function of the reverse GAN network, λ is used to control the relative importance of the two objective functions, λ is a preset value. The overall network task is classified as de-optimized as equation (4):
here, when training the SR-cycleGAN network, 300 sets of raw data may be selected to divide the data into a training set, a validation set, and a test set according to a ratio of 0.8:0.1:0.1, where a sparsely sampled image is used as an input and a fully sampled image (668×668 pixels) is used as an output (i.e., ground trunk). In training, an Adam optimization algorithm is adopted to update network parameters, and the initial learning rate is set to be 0.0001.
Here, the network is adopted to reconstruct images, so that the interested region can be rapidly and continuously imaged, and the acquisition speed of the photoacoustic imaging system is improved while a certain spatial resolution is maintained.
Here, in the conventional imaging method, the sample is scanned and imaged based on the gaussian beam, when the gaussian beam is compared, the spatial light modulator 22 is turned off, the linear polarized gaussian light can be directly transmitted to the scanning array mirror 24 and the objective lens 25 without modulating the linear polarized gaussian light, and because the focusing depth of the gaussian beam is smaller, the three-dimensional moving platform 26 carrying the object needs to be moved along the Z-axis direction to find the focus for two-dimensional sparse synchronous scanning and acquisition, and in the imaging process, the imaging sensitivity is poor due to the limitation of the depth of field of the gaussian beam. In the invention, the Airy light beam can be generated by carrying out phase modulation on linear polarized Gaussian light by the spatial light modulator 22 and Fourier transformation of the lens 25, the Airy light beam is used for carrying out sparse sampling on an object, the sparse image data is obtained and super-resolution reconstruction processing is carried out to recover the photoacoustic microscopic original image, compared with the traditional Gaussian light beam, the Airy light beam has longer focusing length and smaller focusing size, therefore, a high-resolution image with large depth of field can be obtained through two-dimensional synchronous scanning acquisition reconstruction, and the depth of field can keep the transverse resolution better than 4 mu m within the range of 926 mu m. In addition, the main lobe energy of the Airy light can reach more than 50%, so that compared with the Bessel light beam without diffraction, the imaging sensitivity and contrast can be improved.
Fig. 5 is an effect schematic diagram of a photoacoustic microimaging system based on Airy beam joint sparse sampling according to an embodiment of the present invention. In order to characterize the optical characteristics of the modulated light beam, a CMOS camera (Thorlabs) is placed on a back focal plane where the excitation light beam is focused by an objective lens, the CMOS camera is fixedly arranged on the lower end face of the three-dimensional moving platform, the three-dimensional moving platform is moved at a constant speed along the Z-axis direction, a layer of XY-plane light spot image is captured by the CMOS camera once, a three-dimensional stacked image is obtained by overlapping the captured multi-layer XY images, the three-dimensional stacked image is projected on the XZ-plane as a final depth-direction light spot image, the light spot image contains the light spot shape of the current excitation light, airy light spot images with different modulation parameters modulated by a spatial light modulator are obtained, and compared with Gaussian light spot images which are not modulated by the spatial light modulator, wherein the comparison result is shown as (d), (e) and (f) in fig. 5. Specifically, when the transverse scale factors of the gaussian beam and the airy beam with different cubic phase parameters are 3.5 and 1.8, the gaussian beam, the airy beam #1 and the airy beam #2 are marked respectively. Without phase modulation, the measured spot size (full width at half maximum) of the gaussian beam captured at its focal plane through the objective lens is about 11 μm. When the cubic phase parameter is loaded onto the spatial light modulator, the finite energy Airy beam can be captured at the Fourier focal plane, as shown in (e), (f), (h), (i), (k) of FIG. 5, the intensity distribution is a multi-lobe distribution, while the main lobe has a large portion of the total beam energy, which is consistent with the relevant simulation results. Both Airy beams with limited energy exhibit lower diffraction at different lateral scale factors. The main lobe sizes of the airy beam measured at the focus point are about 5 μm and about 3 μm, respectively, in the phase diagram of airy #1 and the phase diagram of airy #2. The attenuation factors of airy #1 and #2 are 0.0011, which determines the attenuation rate of the lateral side lobes and the direction of beam propagation. The light spot is obtained by taking 1/e of the light intensity in the image, which is larger than the maximum light intensity, as a threshold value, the length of the light spot of the Gaussian light in the Z-axis direction is about 58 mu m, the focal length of the Airy light #1 beam is about 11 times of the Gaussian light under the same wavelength, and the focal length of the Airy light #2 beam is about 16 times of the Gaussian light under the same wavelength. From this, it is found that the focal length of the airy ray is much longer than that of the gaussian ray in the Z-axis direction, and the main lobe size of airy ray #2 is smaller than Yu Aili ray #1, but the energy carried by the airy ray is 66% and 42% larger than airy ray #1, respectively.
The invention also provides a photoacoustic microscopic imaging method based on Airy light beam combined sparse sampling, which is applied to the photoacoustic microscopic imaging system, as shown in fig. 6, and comprises the following steps:
s101, generating a nanosecond pulse Gaussian beam, and processing the nanosecond pulse Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam.
S102, performing polarization adjustment, phase modulation, filtering, focusing and Fourier transform processing on the first standard linear polarization Gaussian beam to obtain an Airy beam excited to an object.
S103, when Gaussian beams are polarized according to a second standard line, two-dimensional sparse scanning sampling is conducted on an object excited with Airy beams according to a two-dimensional triangular Lissajous track, and sparse photoacoustic signals are obtained.
S104, receiving the sparse photoacoustic signal, and carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image.
S105, performing image reconstruction on the sparse image by adopting a pre-trained super-resolution reconstruction network to obtain a high-resolution photoacoustic microscopic original image of the object.
Aiming at the problem of mutual restriction among spatial resolution, depth of field, detection sensitivity and imaging speed in the current photoacoustic microscopy imaging, on one hand, the diffraction-free Airy light beam obtained by the light beam generating module and the light beam shaping module has the characteristics of longer focal depth and better focusing effect, and the main lobe of the diffraction-free Airy light beam can carry relatively more energy (about 50%), so that the range of axial imaging of an object (such as biological tissue) can be remarkably expanded, thereby realizing large depth of field and high-sensitivity photoacoustic microscopy imaging, and obtaining a high-resolution imaging image with unchanged depth-direction resolution; on the other hand, the invention carries out two-dimensional sparse scanning sampling on the object excited with the Airy light beam by using a two-dimensional triangular Lissajous track, and carries out image reconstruction by the pre-trained super-resolution reconstruction network provided by the invention after processing the scanning sampling signal, thereby meeting the imaging requirements of high-efficiency and repeatable monitoring. Therefore, the invention can realize the rapid imaging monitoring with large depth of field and high sensitivity on objects (such as physiological and pathological activities).
The foregoing is a further detailed description of the invention in connection with the preferred embodiments, and it is not intended that the invention be limited to the specific embodiments described. It will be apparent to those skilled in the art that several simple deductions or substitutions may be made without departing from the spirit of the invention, and these should be considered to be within the scope of the invention.
Claims (10)
1. Photoacoustic microscopic imaging system based on Airy light beam joint sparse sampling, which is characterized by comprising:
the light beam generation module is used for generating a nanosecond pulse type Gaussian beam, and processing the nanosecond pulse type Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam;
the beam shaping module is used for carrying out polarization adjustment, phase modulation, filtering, focusing and Fourier transformation on the first standard linear polarization Gaussian beam to obtain an Airy beam excited to an object;
the system control and sparse scanning module is used for carrying out two-dimensional sparse scanning sampling on the object excited with the Airy light beam by using a two-dimensional triangular lissajous track when the second standard linear polarized Gaussian light beam is received, so as to obtain a sparse photoacoustic signal;
the signal receiving and processing module is used for receiving the sparse photoacoustic signal and carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image;
the image reconstruction module is used for carrying out image reconstruction on the sparse image by adopting a pre-trained super-resolution reconstruction network to obtain a high-resolution photoacoustic microscopic original image of the object; wherein the super-resolution reconstruction network comprises: two discrimination networks, an up-sampling network and a down-sampling network; the up-sampling network and one discrimination network form a forward GAN network, and the down-sampling network and the other discrimination network form a reverse GAN network.
2. The ai-beam joint sparse sampling based photoacoustic microscopy imaging system of claim 1, wherein the upsampling network comprises, in order: a first convolution layer, an activation layer, a residual sub-network, a sub-pixel convolution layer, and a second convolution layer; the downsampling network comprises, in order: the first convolution layer, the active layer, the residual sub-network, a first downsampled convolution layer, a second downsampled convolution layer, and the second convolution layer; the two discrimination networks are discrimination networks in the super-resolution generation countermeasure network.
3. The ai-beam joint sparse sampling based photoacoustic microscopy imaging system of claim 1, wherein the system control and sparse scanning module comprises: the system comprises a photoelectric detector, a scanning galvanometer, a data acquisition card and a system control module;
the photoelectric detector is used for synchronously sending a trigger signal to the scanning galvanometer and the data acquisition card when receiving the second standard linear polarization Gaussian beam;
the data acquisition card and the system control module are used for generating two-dimensional triangular waveforms at preset frequency and respectively sending the two triangular waveforms to the two vibrating mirrors of the scanning vibrating mirror when the trigger signal is received;
the scanning galvanometer is used for carrying out two-dimensional sparse scanning sampling on the object according to the two-dimensional triangular waveforms received by the two galvanometers and the two-dimensional triangular lissajous tracks to obtain the sparse photoacoustic signal.
4. The ai-beam joint sparse sampling-based photoacoustic microscopy imaging system of claim 1, wherein the beam generation module comprises, in order:
a nanosecond pulse laser for generating a nanosecond pulse gaussian beam;
the polarizer is used for carrying out polarization adjustment on the nanosecond pulse Gaussian beam to obtain a standard linear polarization Gaussian beam;
a polarizing beam splitter for dividing the standard linearly polarized gaussian beam into a first standard linearly polarized gaussian beam and a second standard linearly polarized gaussian beam according to a preset ratio, and taking the second standard linearly polarized gaussian beam as a second standard linearly polarized gaussian beam;
the diaphragm is used for optimizing the first standard linear polarization Gaussian beam to obtain an optimized first standard linear polarization Gaussian beam;
and the beam expanding lens group is used for expanding the optimized first standard linear polarization Gaussian beam to obtain the first standard linear polarization Gaussian beam.
5. The system for photoacoustic microscopy based on Airy beam joint sparse sampling of claim 4, wherein the beam expanding lens group comprises, in order: the first converging lens, the first reflecting mirror, the first small hole and the second converging lens;
the first small hole is used for filtering other stray light in the standard linear polarization Gaussian beam passing through the first converging lens.
6. The system for photoacoustic microscopy based on Airy beam joint sparse sampling of claim 1, wherein the beam shaping module comprises, in order: the device comprises a half-wave plate, a spatial light modulator, a beam shrinking lens group, a scanning array mirror, an objective lens and a three-dimensional moving platform;
the half-wave plate is used for adjusting the polarization direction of the first standard linear polarization Gaussian beam;
the spatial light modulator is used for carrying out phase modulation on the polarized first standard linear polarization Gaussian beam according to preset cube phase modulation parameters to obtain a modulated beam;
the beam shrinking lens group is used for carrying out beam shrinking and filtering treatment on the modulated light beam;
the objective lens is used for focusing and Fourier transforming the light beam passing through the scanning array mirror to obtain the Airy light beam;
the three-dimensional moving platform is used for placing the object and controlling the object to be positioned at the focus of the objective lens through the movement of the three-dimensional moving platform in the horizontal and/or vertical directions.
7. The optical acoustic microscopic imaging system based on Airy beam joint sparse sampling of claim 6, wherein the spatial light modulator is loaded with phase blazed gratings of different grating periods, and the spatial light modulator is further configured to diffract the modulated light beam to different diffraction angles to separate a zero-order light spot and a first-order light spot;
the beam shrinking lens group sequentially comprises: a third converging lens, a second aperture and a fourth converging lens;
and the second small hole is used for filtering the zero-order light spot.
8. The Airy beam joint sparse sampling based photoacoustic microscopy imaging system of claim 6, wherein the beam shaping module further comprises: a third mirror and a fourth mirror; the third reflecting mirror and the fourth reflecting mirror are sequentially positioned between the beam shrinking lens group and the scanning array mirror;
the third reflector is used for reflecting the light beam passing through the beam shrinking lens group to the fourth reflector;
the fourth reflector is used for reflecting the light beam reflected by the third reflector to the scanning array mirror.
9. The ai-beam joint sparse sampling based photoacoustic microscopy imaging system of claim 1, wherein the signal receiving and processing module comprises:
an ultrasonic transducer for receiving the sparse photoacoustic signal and converting the sparse photoacoustic signal into an electrical signal;
and the amplifier is used for amplifying the electric signal to obtain the amplified sparse photoacoustic signal.
10. Photoacoustic microscopic imaging method based on Airy light beam combined sparse sampling, which is applied to a photoacoustic microscopic imaging system according to any one of claims 1 to 9, and comprises the following steps:
generating a nanosecond pulse Gaussian beam, and processing the nanosecond pulse Gaussian beam to obtain a first standard linear polarization Gaussian beam and a second standard linear polarization Gaussian beam;
performing polarization adjustment, phase modulation, filtering, focusing and Fourier transformation on the first standard linear polarization Gaussian beam to obtain an Airy beam excited to an object;
when the Gaussian beam is polarized according to the second standard line, carrying out two-dimensional sparse scanning sampling on the object excited with the Airy beam by using a two-dimensional triangular Lissajous track to obtain a sparse photoacoustic signal;
receiving the sparse photoacoustic signal, and carrying out maximum projection according to the sparse photoacoustic signal to obtain a sparse image;
and adopting a pre-trained super-resolution reconstruction network to reconstruct the image of the sparse image to obtain a high-resolution photoacoustic microscopic original image of the object.
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| CN116879180A (en) * | 2023-09-08 | 2023-10-13 | 之江实验室 | Photoacoustic microimaging system and method based on radial polarization modulation beam focusing |
| CN117092033A (en) * | 2023-10-19 | 2023-11-21 | 深圳湾实验室 | All-optical photoacoustic imaging system of asymmetric Airy light |
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| CN116879180A (en) * | 2023-09-08 | 2023-10-13 | 之江实验室 | Photoacoustic microimaging system and method based on radial polarization modulation beam focusing |
| CN116879180B (en) * | 2023-09-08 | 2023-12-01 | 之江实验室 | Photoacoustic microimaging system and method based on radial polarization modulation beam focusing |
| CN117092033A (en) * | 2023-10-19 | 2023-11-21 | 深圳湾实验室 | All-optical photoacoustic imaging system of asymmetric Airy light |
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