CN114129132A - Large-field-of-view high-speed photoacoustic microscopic imaging device and method - Google Patents
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Abstract
The invention discloses a large-view-field high-speed photoacoustic microscopic imaging device and method. The device comprises a laser source assembly, a light beam transmission assembly, a light beam scanning assembly, a reflective imaging port assembly and a computer. The method uses nanosecond pulse laser emitted by a pulse laser transmitted by a sheet optical fiber bundle as excitation light, uses a micro-lens array to realize light focusing, simultaneously excites photoacoustic signals on a single radius of an imaging area, uses a line focusing ultrasonic transducer array to simultaneously detect the photoacoustic signals on the single radius, and combines a radius rotation scanning method to realize three-dimensional imaging. Expanding the visual field of the photoacoustic microscopic imaging to centimeter magnitude by radius rotation scanning; by means of simultaneous laser, simultaneous acquisition and reconstruction of photoacoustic signals, imaging time is shortened to the second level, high-speed imaging under the condition of a large field of view is achieved, and conditions are provided for animal organ dimension photoacoustic imaging and large animal photoacoustic imaging.
Description
Technical Field
The invention belongs to the field of medical equipment and methods, and particularly relates to a large-field-of-view high-speed photoacoustic microscopic imaging device and method.
Background
The photoacoustic microimaging is a novel optical imaging method which irradiates a focused laser on an imaging sample, excites ultrasonic waves, detects the ultrasonic waves to obtain light absorption information in a certain depth in the sample, and realizes three-dimensional imaging by scanning a focused light beam. Due to the strong absorption of visible light by hemoglobin, photoacoustic microscopy is widely used to reveal the structural and functional changes of three-dimensional vascular networks of various models of animals and humans.
At present, the common realization method disclosed at home and abroad photoacoustic microimaging is to use an objective lens or a scanning lens to focus pulse laser to excite a photoacoustic signal, use a point focusing ultrasonic transducer to detect the signal and use a two-dimensional linear motor to scan a photoacoustic microimaging system or a target to obtain an image. The method is limited by a screw rod and sliding block structure of a linear motor, the imaging speed of a large-size target is low, the system volume is large, the relative movement exists between the system and the target, and the like, and the imaging range is generally 1 multiplied by 1 cm2On the other hand, the imaging time depends on the repetition rate of the pulse laser, and the imaging time is about 100 seconds using a laser repetition rate of 10 kHz.
In order to solve the problems, a plurality of research teams at home and abroad propose to scan a focused laser beam by using a scanning galvanometer and detect a signal by using a flat-field ultrasonic transducer; or a water immersion type micro scanning galvanometer or a polygonal scanner is used for scanning a focused laser beam and a point focusing ultrasonic transducer ultrasonic sound field simultaneously, and a pulse laser with high repetition frequency is combined, so that rapid imaging in a slightly large range is realized to a certain extent. However, these methods either sacrifice the signal-to-noise ratio of signal detection or are affected by factors such as liquid damping, and thus do not fundamentally solve the problem.
Another solution is to use a line-focusing ultrasonic transducer to detect photoacoustic signals, use an optical scanning galvanometer to scan along the ultrasonic focusing line, rotate the ultrasonic transducer along the center after completing one scan and repeat the above steps until the rotation angle reaches 360 degrees to cover the circular imaging area, and the imaging time is 400 seconds in the imaging range of 4 cm in diameter.
Although the solution described above achieves large field-of-view imaging and obtains a higher signal-to-noise ratio by the line focus ultrasound transducer, the contradictory relationship between the number of scanning points and the scanning time under large field-of-view imaging still cannot be solved, and meanwhile, due to the limitation of the high-frequency ultrasound transducer, a single ultrasound transducer is difficult to achieve imaging in a larger range. These problems limit the application of photoacoustic microscopy in clinical and biomedical research.
Disclosure of Invention
Technical problem to be solved
Aiming at the problems of small field of view and low speed of the existing photoacoustic microscopic imaging method, the invention aims to provide a novel photoacoustic microscopic imaging device and method combining a microlens array focusing mechanism, a line focusing array ultrasonic transducer detecting mechanism and a radius rotating scanning mechanism, which can perform high-speed imaging under the imaging condition of a large field of view, improve the field of view of the existing photoacoustic microscopic imaging, shorten the imaging time of the existing photoacoustic microscopic imaging, meet the requirements of high speed and large field of view, and provide a novel imaging device for the biomedical imaging requirement.
(II) technical scheme
The invention provides a large-field-of-view high-speed photoacoustic microscopic imaging device and method for solving the technical problem, and the specific technical scheme is as follows.
A large-field-of-view high-speed photoacoustic microscopic imaging device is characterized in that: the device comprises a laser source component, a light beam transmission component, a light beam scanning component, a reflective imaging port component and a computer;
the laser source assembly comprises a pulse laser for emitting pulse laser to an imaging target;
the light beam transmission assembly comprises a spatial optical filter, a convex lens and a sheet-shaped optical fiber bundle and is used for realizing coaxial convergence, shaping and transmission of pulse laser;
the light beam scanning assembly comprises an optical fiber rotating motor, an optical fiber rotating gear set, an optical fiber rotating motor controller and a linearly arranged micro-lens array, and is used for realizing the scanning and focusing of pulse laser;
the reflection type imaging port assembly comprises an optical-acoustic signal coaxial coupling device, an energy converter rotating motor, an energy converter rotating gear set, an energy converter rotating motor controller, a line focusing ultrasonic energy converter array and a multi-channel amplifying filtering and collecting system, and is used for realizing excitation, transmission and collection of optical-acoustic signals; the optical-acoustic signal coaxial coupling device is internally provided with a quartz glass cover glass which is fixed at an inclination angle of 45 degrees, deionized water is filled in the quartz glass cover glass to serve as coupling liquid, and the surface of the quartz glass cover glass is sealed by a transparent film; light output by the sheet-shaped optical fiber bundle passes through the light-sound signal coaxial coupling device after being focused by the linearly arranged micro-lens array and then irradiates on an imaging sample, and focusing light spots are linearly arranged and coincide with a focal region of the line focusing ultrasonic transducer array; the imaging sample absorbs light energy to generate photoacoustic signals, enters the light-sound signal coaxial coupling device, and is reflected to the surface of the line focus ultrasonic transducer array by the quartz glass cover glass to realize detection;
the computer is used for controlling the synchronization of the optical fiber rotating motor in the light beam scanning assembly and the transducer rotating motor in the reflective imaging port assembly; and meanwhile, the method is also used for photoacoustic signal reconstruction and image processing.
More specifically, the laser source assembly emits pulsed laser light to an imaging target, the pulse repetition frequency is 10 kHz, and the operating wavelength of the pulsed laser is 532 nm.
More specifically, the optical beam transmission assembly operates such that the pulsed laser light output by the pulsed laser is emitted as spatial light, first through a spatial optical filter and then coupled into the circular input end of the slab fiber bundle through a convex lens.
More specifically, the light beam scanning assembly works in a mode that pulse laser is output from an output end of a sheet-shaped optical fiber bundle, optical fibers in the optical fiber bundle are specially designed and arranged to enable output light spots to be rectangular, the output light is transmitted to a linearly arranged micro lens array, the output light passes through an optical-acoustic signal coaxial coupling device and then converges at a focus to form a line to irradiate on an imaging sample, and an optical fiber rotating motor is used for driving the sheet-shaped optical fiber bundle and the micro lens array to rotate.
More specifically, the reflective imaging port assembly works in such a way that the refractive index of deionized water in the optical-acoustic signal coaxial coupling device in the assembly is similar to that of a quartz glass cover glass, so that a pulse light beam directly penetrates through the device and irradiates on an imaging sample; absorbing light energy by tissues of an imaging sample to generate ultrasonic waves, reflecting the excited backward ultrasonic waves to a line focusing ultrasonic transducer array by the quartz glass cover glass due to the acoustic impedance difference between deionized water and the quartz glass cover glass, converting the ultrasonic waves into electric signals, amplifying and filtering the electric signals, and storing the electric signals in a computer; during image acquisition, after signal acquisition on one line is completed, the transducer rotating motor respectively drives the sheet-shaped optical fiber bundle, the micro lens array and the line focusing ultrasonic transducer array to rotate, so that output light of the sheet-shaped optical fiber bundle, a light focal line after passing through the micro lens array and an acoustic focal line after the line focusing ultrasonic transducer array is reflected by the quartz glass cover glass coincide, one end of the line rotates by a preset angle (such as 0.018 degrees) to perform acquisition, the radius of the line is shown to rotate along the center of a circle on an imaging surface until the rotation angle reaches 360 degrees, and the acquisition is completed.
A large-field-of-view high-speed photoacoustic microscopic imaging method is characterized in that a photoacoustic image is obtained by adopting any one of the large-field-of-view high-speed photoacoustic microscopic imaging devices, and the method comprises the following steps:
step S1, placing the experimental animal which is depilated and cleaned on an imaging table, smearing an ultrasonic coupling agent on the part to be imaged to be tightly attached to the transparent sealing film of the optical-acoustic signal coaxial coupling device;
step S2, the pulse laser emits pulse laser, which passes through the space optical filter, the convex lens, the sheet optical fiber bundle and the micro lens array in sequence, the focused light beam passes through the optical-acoustic signal coaxial coupling device, irradiates on the imaging part of the experimental animal, and excites the photoacoustic signal; the photoacoustic signal penetrates through a transparent sealing film on the surface of the light-sound signal coaxial coupling device in the form of mechanical wave, enters the interior, is transmitted in deionized water, is reflected to the surface of the line focusing ultrasonic transducer array by a quartz glass cover glass fixed at an inclination angle of 45 degrees and is converted into an electric signal; wherein, the photoacoustic signal generated at each optical focus position of the micro lens array contains a depth signal of the point, which is called as an 'A line signal', the focuses of the micro lens array are arranged in sequence, and the generated two-dimensional data containing the depth is called as 'B scanning data';
step S3, after obtaining the "B scan data" at the initial angle, the fiber rotation motor and the transducer rotation motor respectively drive the sheet-like fiber bundle, the microlens array and the line focus ultrasound transducer array to rotate a preset angle (e.g. 0.018 degree) along one side through the fiber rotation gear set and the transducer rotation gear set, which is expressed that the focused light beam and the ultrasound focal line on the imaging window rotate around one side as an axis and rotate around the same angle (0.018 degree) at the same time as a radius to keep synchronization;
step S4, continuously repeating the steps S2 and S3 until the rotation angle reaches 360 degrees, and completing the scanning and data acquisition of one-time imaging;
step S5, carrying out image reconstruction on the acquired data; reconstructing the obtained B scanning data by using a filtering back projection algorithm to obtain two-dimensional images with depth on each radius, and improving the resolution by combining light focusing and positioning; mapping the two-dimensional image collected by rotation to a rectangular coordinate system by using a coordinate system conversion algorithm, and reconstructing the collected two-dimensional image into a three-dimensional image; and finally, optimizing the imaging effect by utilizing three-dimensional image filtering.
(III) advantageous effects
Compared with the prior art, the invention has obvious and positive technical effects, and the beneficial effects are at least reflected in the following aspects.
(1) The photoacoustic microimaging method and the photoacoustic microimaging device adopt the rotating motor to control the focused light beam and the ultrasonic transducer array to scan, the imaging range depends on the effective detection range of the array ultrasonic transducer and the size of the microlens array, and the photoacoustic microimaging with an oversized view field (the imaging range is larger than the diameter of 5 cm) can be realized.
(2) The photoacoustic microimaging method and the photoacoustic microimaging device provided by the invention have the advantages that the micro-lens array is used for focusing laser, the array ultrasonic transducer is used for detecting signals, photoacoustic signals at all positions on a line can be obtained at the same time for reconstruction, the imaging speed of the photoacoustic microimaging method and the photoacoustic microimaging device depends on the step length of rotary scanning and the pulse repetition frequency of a pulse laser, and high-speed (reaching the second level) photoacoustic microimaging can be realized.
Drawings
Fig. 1 is a schematic system structure diagram of a large-field-of-view high-speed photoacoustic microscopic imaging device according to the present invention.
Fig. 2 is a basic principle schematic diagram of the large-field-of-view high-speed photoacoustic microscopic imaging method of the invention.
Detailed Description
The invention provides a large-field-of-view high-speed photoacoustic microscopic imaging device and method for solving the technical problem. The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings of the specification.
The invention provides a large-field-of-view high-speed photoacoustic microscopic imaging system, and particularly relates to a device which comprises a laser source assembly 1, a light beam transmission assembly 2, a light beam scanning assembly 3, a reflective imaging port assembly 4 and a computer 5, wherein the light beam transmission assembly 2 is arranged on the laser source assembly.
As shown in fig. 1, the light beam transmission assembly includes a spatial optical filter 2-1, a convex lens 2-2, and a sheet-like optical fiber bundle 2-2.
As shown in FIG. 1, the beam scanning assembly includes a micro lens array 3-1, a motor controller 3-2, a fiber rotating motor 3-3, and fiber rotating gear sets 3-4, 3-5.
As shown in FIG. 1, the reflective imaging port assembly comprises an optical-acoustic signal coaxial coupling device (a lower sealing cover glass 4-1, a quartz glass cover glass 4-2 obliquely installed at 45 degrees, a surface transparent sealing film 4-3 and filled deionized water), a line focusing ultrasonic transducer array 4-4, a transducer rotating motor 4-5, transducer rotating gear sets 4-6 and 4-7, a signal amplifier 4-8, a band-pass filter 4-9 and a data acquisition card 4-10.
Specifically, pulse laser emitted by the laser source assembly 1 is coupled into a sheet-shaped optical fiber bundle 2-3 through a spatial optical filter 2-1 and a convex lens 2-2, the pulse laser output by the optical fiber bundle is transmitted to a linearly arranged micro lens array 3-1, passes through a lower sealing cover glass 4-1 of an optical-acoustic signal coaxial coupling device, a quartz glass cover glass 4-2 obliquely arranged at an angle of 45 degrees and a surface transparent sealing film 4-3, and then is converged into a line at a focal point to irradiate on an imaging sample. The photoacoustic signal enters the interior of the linear focusing ultrasonic transducer array 4-4 through the transparent sealing film 4-3, is transmitted in deionized water and is reflected to the surface of the linear focusing ultrasonic transducer array 4-4 by the quartz glass cover glass 4-2 which is fixed at an inclination angle of 45 degrees. The signals pass through a signal amplifier 4-8, a band-pass filter 4-9 and a data acquisition card 4-10 in sequence, are stored in a computer 5 and are reconstructed.
After the acquisition of one imaging plane is finished, the motor controller 3-2 controls the two rotating motors 3-3 and 4-5 to respectively drive the sheet-shaped optical fiber bundle 2-2, the microlens array 3-1 and the line focusing ultrasonic transducer array 4-4 to rotate by a preset angle (such as 0.018 degrees) through the transmission gear sets 3-4, 3-5, 4-6 and 4-7 to reach the next imaging plane for continuous acquisition.
The basic principle of the large-field-of-view high-speed photoacoustic microimaging method is shown in fig. 2, and the method mainly uses key components such as a sheet-shaped optical fiber bundle 1, a linearly-arranged microlens array 2, a line-focus ultrasonic transducer array 5 and the like, and specifically comprises the following steps:
(1) exciting and detecting photoacoustic signals, wherein the sheet-shaped optical fiber bundle 1 transmits pulse laser emitted by a pulse laser to the surface of the micro-lens array 2, pulse light focuses after passing through the micro-lens array 2 are linearly arranged and irradiated on a sample and irradiated on an imaging part of an experimental animal, and the photoacoustic signals are excited; the photoacoustic signal is converted into an electric signal on the surface of the on-line focusing ultrasonic transducer array 5; the photoacoustic signal generated at each optical focus position of the micro lens array comprises a depth signal of the point, and the focuses of the micro lens array are sequentially arranged to generate two-dimensional data comprising the depth;
(2) after the photoacoustic signal data of a certain plane is acquired, rotating a preset angle (such as 0.018 degrees) by taking one side 6 of an imaging surface as an axis and the length of the imaging surface as a radius;
(3) continuously repeating the steps (1) and (2) until the rotation angle reaches 360 degrees;
(4) and (5) image reconstruction. Obtaining a two-dimensional image with a single imaging surface containing depth by using a filtering back projection algorithm, and improving resolution by combining light focusing positioning; mapping the two-dimensional image to a rectangular coordinate system by using a coordinate system conversion algorithm, and reconstructing the two-dimensional image into a three-dimensional image; and finally, optimizing the imaging effect by utilizing three-dimensional image filtering.
The embodiments described in this application are only intended to illustrate the main idea of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (6)
1. A large-field-of-view high-speed photoacoustic microscopic imaging device is characterized in that: the device comprises a laser source component, a light beam transmission component, a light beam scanning component, a reflective imaging port component and a computer;
the laser source assembly comprises a pulse laser for emitting pulse laser to an imaging target;
the light beam transmission assembly comprises a spatial optical filter, a convex lens and a sheet-shaped optical fiber bundle and is used for realizing coaxial convergence, shaping and transmission of pulse laser;
the light beam scanning assembly comprises an optical fiber rotating motor, an optical fiber rotating gear set, an optical fiber rotating motor controller and a linearly arranged micro-lens array, and is used for realizing the scanning and focusing of pulse laser;
the reflection type imaging port assembly comprises an optical-acoustic signal coaxial coupling device, an energy converter rotating motor, an energy converter rotating gear set, an energy converter rotating motor controller, a line focusing ultrasonic energy converter array and a multi-channel amplifying filtering and collecting system, and is used for realizing excitation, transmission and collection of optical-acoustic signals; the optical-acoustic signal coaxial coupling device is internally provided with a quartz glass cover glass which is fixed at an inclination angle of 45 degrees, deionized water is filled in the quartz glass cover glass to serve as coupling liquid, and the surface of the quartz glass cover glass is sealed by a transparent film; light output by the sheet-shaped optical fiber bundle passes through the light-sound signal coaxial coupling device after being focused by the linearly arranged micro-lens array and then irradiates on an imaging sample, and focusing light spots are linearly arranged and coincide with a focal region of the line focusing ultrasonic transducer array; the imaging sample absorbs light energy to generate photoacoustic signals, enters the light-sound signal coaxial coupling device, and is reflected to the surface of the line focus ultrasonic transducer array by the quartz glass cover glass to realize detection;
the computer is used for controlling the synchronization of the optical fiber rotating motor in the light beam scanning assembly and the transducer rotating motor in the reflective imaging port assembly; and meanwhile, the method is also used for photoacoustic signal reconstruction and image processing.
2. The large-field-of-view high-speed photoacoustic microscopy imaging apparatus according to claim 1, wherein: the laser source component emits pulse laser to an imaging target, the pulse repetition frequency is 10 kHz, and the working wavelength of the pulse laser is 532 nm.
3. The large-field-of-view high-speed photoacoustic microscopy imaging apparatus according to claim 1, wherein: the working mode of the light beam transmission component is as follows, pulse laser output by a pulse laser is emitted in the form of space light, and the pulse laser firstly passes through a space optical filter and then enters a circular input end of a sheet-shaped optical fiber bundle through the coupling of a convex lens.
4. The large-field-of-view high-speed photoacoustic microscopy imaging apparatus according to claim 1, wherein: the working mode of the light beam scanning assembly is as follows, pulse laser is output from the output end of the sheet-shaped optical fiber bundle, optical fibers in the optical fiber bundle are arranged through specific design, so that output light spots are rectangular, the output light is transmitted to the linearly arranged micro lens array, the output light passes through the light-sound signal coaxial coupling device and then converges at a focus to form a line to irradiate on an imaging sample, and the optical fiber rotating motor is used for driving the sheet-shaped optical fiber bundle and the micro lens array to rotate.
5. The large-field-of-view high-speed photoacoustic microscopy imaging apparatus according to claim 1, wherein: the reflective imaging port assembly works in a mode that the refractive index of deionized water in the optical-acoustic signal coaxial coupling device in the assembly is approximate to that of a quartz glass cover glass, so that a pulse light beam directly penetrates through the device to irradiate on an imaging sample; absorbing light energy by tissues of an imaging sample to generate ultrasonic waves, reflecting the excited backward ultrasonic waves to a line focusing ultrasonic transducer array by the quartz glass cover glass due to the acoustic impedance difference between deionized water and the quartz glass cover glass, converting the ultrasonic waves into electric signals, amplifying and filtering the electric signals, and storing the electric signals in a computer; during image acquisition, after signal acquisition on one line is completed, the optical fiber rotating motor respectively drives the sheet-shaped optical fiber bundle, the micro lens array and the line focusing ultrasonic transducer array to rotate, so that output light of the sheet-shaped optical fiber bundle, a light focal line after passing through the micro lens array and a sound focal line after the line focusing ultrasonic transducer array is reflected by the quartz glass cover glass coincide, and one end of the line rotates by a preset angle (such as 0.018 degrees) to perform acquisition, wherein the radius of the acquisition is represented on an imaging surface and the rotation is performed along the center of a circle until the rotation angle reaches 360 degrees, and the acquisition is completed.
6. A large-field-of-view high-speed photoacoustic microscopy imaging method, characterized in that photoacoustic imaging is obtained by using the large-field-of-view high-speed photoacoustic microscopy imaging apparatus according to any one of claims 1 to 5, and the steps are as follows:
step S1, placing the experimental animal which is depilated and cleaned on an imaging table, smearing an ultrasonic coupling agent on the part to be imaged to be tightly attached to the transparent sealing film of the optical-acoustic signal coaxial coupling device;
step S2, the pulse laser emits pulse laser, which passes through the space optical filter, the convex lens, the sheet optical fiber bundle and the micro lens array in sequence, the focused light beam passes through the optical-acoustic signal coaxial coupling device, irradiates on the imaging part of the experimental animal, and excites the photoacoustic signal; the photoacoustic signal penetrates through a transparent sealing film on the surface of the light-sound signal coaxial coupling device in the form of mechanical wave, enters the interior, is transmitted in deionized water, is reflected to the surface of the line focusing ultrasonic transducer array by a quartz glass cover glass fixed at an inclination angle of 45 degrees and is converted into an electric signal; wherein, the photoacoustic signal generated at each optical focus position of the micro lens array contains a depth signal of the point, which is called as an 'A line signal', the focuses of the micro lens array are arranged in sequence, and the generated two-dimensional data containing the depth is called as 'B scanning data';
step S3, after obtaining the "B scan data" at the initial angle, the fiber rotation motor and the transducer rotation motor respectively drive the sheet-like fiber bundle, the microlens array and the line focus ultrasound transducer array to rotate a preset angle (e.g. 0.018 degree) along one side through the fiber rotation gear set and the transducer rotation gear set, which is expressed that the focused light beam and the ultrasound focal line on the imaging window rotate around one side as an axis and rotate around the same angle (0.018 degree) at the same time as a radius to keep synchronization;
step S4, continuously repeating the steps S2 and S3 until the rotation angle reaches 360 degrees, and completing the scanning and data acquisition of one-time imaging;
step S5, carrying out image reconstruction on the acquired data; reconstructing the obtained B scanning data by using a filtering back projection algorithm to obtain two-dimensional images with depth on each radius, and improving the resolution by combining light focusing and positioning; mapping the two-dimensional image collected by rotation to a rectangular coordinate system by using a coordinate system conversion algorithm, and reconstructing the collected two-dimensional image into a three-dimensional image; and finally, optimizing the imaging effect by utilizing three-dimensional image filtering.
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