CN111948145A - Bessel beam large-depth-of-field photoacoustic microscopic imaging device and method based on ultrasonic modulation - Google Patents
Bessel beam large-depth-of-field photoacoustic microscopic imaging device and method based on ultrasonic modulation Download PDFInfo
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- CN111948145A CN111948145A CN202010706502.1A CN202010706502A CN111948145A CN 111948145 A CN111948145 A CN 111948145A CN 202010706502 A CN202010706502 A CN 202010706502A CN 111948145 A CN111948145 A CN 111948145A
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Abstract
A large-depth-of-field Bessel beam photoacoustic microscopic imaging device based on ultrasonic modulation comprises a pulse laser, a piezoelectric ceramic round tube, a three-dimensional scanner, a function generator, a digital pulse delayer and a synchronous circuit consisting of a D-type trigger. The pulse laser emits laser pulses, the laser pulses are incident to the piezoelectric ceramic round tube filled with liquid, the function generator applies sinusoidal radio-frequency signals to the piezoelectric ceramic round tube, the piezoelectric ceramic round tube vibrates in the radial direction to generate ultrasonic waves, the refractive index of the liquid in the radial direction is distributed as a zero-order Bessel function, the synchronous circuit synchronizes the light emission of the laser and the refractive index change of the piezoelectric ceramic round tube, so that the pulse laser emits light when the refractive index of the piezoelectric ceramic round tube is changed to be positive and maximum, at the moment, the light beam coming out of the piezoelectric ceramic round tube is a Bessel light beam, and the light beam is focused on a sample through the. The invention uses ultrasonic modulation to generate Bessel beams, realizes the expansion of the depth of field of photoacoustic microscopic imaging and is beneficial to the rapid and large-scale monitoring of physiological activities.
Description
Technical Field
The invention relates to the field of optical imaging, in particular to a large-depth-of-field Bessel beam photoacoustic microscopic imaging device and method based on ultrasonic modulation.
Background
The photoacoustic imaging technology is a biomedical imaging technology which is rapidly developed in recent years, combines high contrast of optical imaging and high penetrability and high resolution of acoustic imaging, and has a biomedical imaging mode with deep imaging depth and high spatial resolution. In recent years, the method has wide application in various fields of biomedicine, such as vascular structure, tumor detection, brain structure and function imaging. The photoacoustic microscopic imaging is an important branch of photoacoustic imaging, has higher spatial resolution, and can realize multi-scale imaging from cells to tissues. In photoacoustic microscopy, the optical focus and the acoustic focus are usually coaxial confocal, and the lateral resolution is determined by the optical focus and the acoustic focus, which can be divided into an optical resolution photoacoustic microscopy imaging system and an acoustic resolution photoacoustic microscopy imaging system. In an acoustic-resolved photoacoustic microscopy imaging system, incident light is weakly focused, the size of the optical focus is much larger than the size of the acoustic focus, and therefore the lateral resolution is determined by the smaller acoustic focus. In an optical resolution photoacoustic microscopy imaging system, an objective lens with a high numerical aperture is generally used for strongly focusing incident light, the size of an optical focus is far smaller than that of an acoustic focus, and the transverse resolution depends on the size of the optical focus. However, in an optical resolution photoacoustic microscopy imaging system, strong focusing of the light beam to obtain high resolution results in a small depth of field of the imaging, and the lateral resolution deteriorates rapidly beyond the optical focus. The smaller imaging depth of field causes the volume imaging speed of the system to be limited, and physiological activities and the like cannot be rapidly monitored in a large range.
To solve this problem, many researchers have proposed different approaches. One relatively common simple method is to use mechanical scanning, but the method has the problems of slow scanning speed, limited precision, introduction of mechanical vibration and the like. Researchers have also obtained twice the depth of field of the image by using illumination from both up and down directions, but this system can only be transmissive, and can only image a transparent or thin sample with a large depth of field. Researchers also utilize the chromatic aberration characteristic of a non-achromatic objective lens and utilize a multi-wavelength laser to generate a plurality of focuses along the axial direction, so that the axial imaging range of the system is improved, but the method sacrifices the functional imaging of the system.
Disclosure of Invention
Based on this, it is necessary to provide a large depth-of-field bessel beam photoacoustic microscopic imaging apparatus and method based on ultrasonic modulation, aiming at the above mentioned problems.
A big depth of field Bessel light beam optoacoustic microscopic imaging device based on ultrasonic modulation is characterized in that: the large-depth-of-field Bessel beam photoacoustic microscopic imaging device based on ultrasonic modulation mainly comprises a pulse laser, a piezoelectric ceramic circular tube, a three-dimensional scanner, a function generator and a digital pulse delayer.
A large-depth-of-field Bessel beam photoacoustic microscopic imaging method based on ultrasonic modulation comprises the following steps:
s1: the pulse laser emits laser pulses, the size of a light spot is shaped to be equivalent to the caliber of the piezoelectric ceramic round tube through the beam expanding system, and then the laser pulses are incident to the piezoelectric ceramic round tube filled with the optical transparent liquid;
s2: applying a sinusoidal radio frequency signal to the piezoelectric ceramic round tube by using a function generator, vibrating the inner wall and the outer wall of the piezoelectric ceramic round tube and generating ultrasonic waves, wherein the ultrasonic waves can periodically modulate the density of local optical transparent liquid so as to change the refractive index distribution of the optical transparent liquid, and the refractive index distribution of the optical transparent liquid in the radial direction is represented as zero-order Bessel function distribution and is synchronously changed with the sinusoidal radio frequency signal;
s3: synchronizing the light emission of the laser and the change of the refractive index of the liquid in the piezoelectric ceramic round tube by using a synchronous circuit, so that the laser emits light on one phase of the change of the refractive index;
the pulse laser can emit light when the refractive index of the liquid in the piezoelectric ceramic round tube becomes positive and maximum by controlling the time delay of the digital pulse delayer, and the radial refractive index distribution of the liquid in the piezoelectric ceramic round tube is zero-order Bessel function distribution at the moment. Thus, the light beam coming out of the piezoelectric ceramic round tube after ultrasonic modulation is a Bessel light beam.
And S4, focusing the obtained bessel beam on the sample through a fifth condenser lens. The generated photoacoustic signal is detected by an acoustic lens, amplified by an amplifier and finally collected by a collecting card.
S5 obtaining three-dimensional data by two-dimensional raster scanning with a three-dimensional scanner
The beam expanding system in S1 is composed of a first condenser lens and a second condenser lens.
Preferably, the piezoelectric ceramic circular tube is polarized and then plated on the inner wall and the outer wall to form an inner electrode and an outer electrode, the inner electrode extends from one end to the outer wall, and the inner electrode and the outer electrode form two gaps on the outer wall and the end face of the other end.
Preferably, the laser pulse vertically enters the circular piezoelectric ceramic tube, and the circular piezoelectric ceramic tube is vertically placed, so that the radial refractive index distribution is not symmetrical due to gravity when the circular piezoelectric ceramic tube is horizontally placed.
Preferably, the optically transparent liquid is silicone oil, and the silicone oil has a kinematic viscosity of 50-150cSt, a refractive index of 1-2, and a sound velocity of 800-1200 m/s.
The refractive index distribution of the optically transparent liquid after ultrasonic modulation described in S2 is
Wherein n is0Is the static refractive index of the medium, csIs the speed of sound in the medium, ω is the frequency of the drive signal, r and t denote position and time, respectively, J0Is a zero order Bessel function, naIs a constant related to omega, the physical properties of the acoustic medium, and the refractive index distribution behaves as a zero order bessel function and changes synchronously with the drive signal over time.
Preferably, the synchronization circuit in S3 is composed of a three-dimensional scanner, a function generator, a D-type trigger, and a digital pulse delay unit, the function generator is used to output a sinusoidal radio frequency signal to drive the circular piezoelectric ceramic tube, the synchronization signal of the sinusoidal radio frequency signal is connected to the clock end of the D-type trigger, the three-dimensional scanner outputs a position synchronization signal every step of horizontal movement, and the position synchronization signal is a TTL square wave signal (the frequency is much less than the frequency of the sinusoidal drive signal of the circular piezoelectric ceramic tube, such as several kilohertz) connected to the D end of the D-type trigger;
according to the characteristic equation of the D-type trigger, the D-type trigger is triggered only when the synchronous signal rises, and the output signal is equal to the input signal; the output end of the D-type trigger is connected with a digital pulse delayer, and the output end of the digital pulse delayer is used for triggering a pulse laser;
the pulse laser can emit light when the refractive index of the liquid in the piezoelectric ceramic round tube changes to be the maximum by adjusting the time delay of the digital pulse delayer. The light beam coming out of the piezoelectric ceramic round tube is a Bessel light beam.
Preferably, the acoustic lens in S4 is composed of an ultrasonic transducer and an acousto-optic coupling prism, and the numerical aperture of the acousto-optic coupling prism is 0.5.
The invention has the advantages and beneficial effects that:
the invention has the advantages that the piezoelectric ceramic round tube generates ultrasonic waves under the drive of sinusoidal radio frequency signals, the radial refractive index distribution of the optical transparent liquid is modulated into zero-order Bessel function distribution by the ultrasonic waves, and then Bessel beams are obtained. The Bessel beam has a non-diffraction characteristic and has a larger depth of field compared with the Gaussian beam. The device will promote the depth of field of optoacoustic microscopic imaging system, and then promote its volume imaging speed, can carry out quick high-resolution monitoring on a large scale to physiological activity better, expand its range of application in biomedicine.
Drawings
Fig. 1 is a schematic view of a large-depth-of-field bessel beam photoacoustic microscopic imaging apparatus based on ultrasonic modulation in an embodiment of the present invention;
FIG. 2 shows the radial distribution of the refractive index of the liquid filled in the piezoelectric ceramic circular tube when the refractive index changes to the positive maximum.
In the figure: 1. a pulsed laser; 2. a first condenser lens; 3. a second condenser lens; 4. a first reflector; 5. a second reflector; 6. a piezoelectric ceramic circular tube; 7. a third condensing lens; 8. a fourth condenser lens; 9. a fifth condenser lens; 10. a function generator; 11. a D-type flip-flop; 12. a digital pulse delayer; 13. an ultrasonic transducer; 14. an acousto-optic coupling prism; 15. a water tank; 16. a sample; 17. a three-dimensional scanner; 18. an amplifier; 19. collecting cards; 20. a workstation.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Example 1
The device and the method of the invention are as follows: FIG. 1 is a schematic view of the structure of the whole set of the image forming apparatus of the present invention. The laser light source is Nd, namely YLF pulse laser 1, and the pulse laser 1 emits laser with the pulse frequency of 1KHz, the wavelength of 523 nanoseconds and the pulse width of 9 nanoseconds. After the laser is emitted, the laser is shaped to a spot diameter of 20mm through a beam expanding system consisting of a first condenser lens 2 and a second condenser lens 3, then the laser beam is converted from horizontal to vertical through a first reflector 4, and then the laser beam is vertically incident into a piezoelectric ceramic circular tube 6. The laser coming out of the piezoelectric ceramic round tube passes through a third condenser lens 7, is reflected by a second reflector 5, enters a fourth condenser lens 8, is finally strongly focused on a sample 16 through a fifth condenser lens 9, generates a Bessel beam in a focal region, and is excited to generate a photoacoustic signal. The generated photoacoustic signals are detected and received by an acousto-optic coupling prism (numerical aperture is 0.5)14 and an ultrasonic transducer 13 (central frequency is 50MHz, olympus), and are acquired by a signal acquisition card 19 with the sampling rate of 500MHz (the number of samples is 1024) after being amplified by an amplifier 18. The collected signals are transmitted to the workstation 20 for analysis. The water bath 15 serves to couple the photoacoustic signals. The three-dimensional scanner 17 is used to adjust the position of the sample 16 in the lateral and axial directions.
The piezoelectric ceramic cylinder 6(PZT-8) is filled with silicone oil (100 cSt). The inner diameter of the piezoelectric ceramic round tube is 16mm, the outer diameter is 20mm, and the length is 20 mm. The refractive index of the silicone oil is 1.403, and the sound velocity is 1000 m/s. When the piezoelectric ceramic round tube is driven by a sinusoidal radio frequency driving signal, the refractive index of the silicon oil shows synchronous change with the driving signal.
The function generator 10 is used for emitting light with the frequency of 707kHz and the peak-to-peak voltage of 10Vp-pThe sinusoidal driving signal drives the piezoelectric ceramic circular tube 6, and meanwhile, the synchronous signal of the driving signal is connected to the clock end of the D-type trigger 11. The three-dimensional scanner 17 outputs a position synchronization signal to the D end of the D-type trigger every time it moves in the transverse direction, and the position synchronization signal is a TTL square wave signal (the frequency is much less than the frequency of the sinusoidal driving signal of the piezoelectric ceramic circular tube, for example, several kilohertz). The output end of the D-type trigger is connected with a digital pulse delayer 12, the digital pulse delayer 12 outputs signals as the trigger signals of the pulse laser 1, so that the pulse laser 1 is synchronous with the driving signals of the piezoelectric ceramic round tube 6, and the laser pulse is synchronous with a certain vibration state of the piezoelectric ceramic round tube 6. Thus, the laser emits light when the refractive index of the piezoelectric ceramic round tube is changed to be positive and maximum by adjusting the delay of the digital pulse delayer 12, and at the moment, the radial refractive index distribution of liquid in the piezoelectric ceramic round tube is zero-order Bessel function distribution, as shown in FIG. 2. Thus, the light beam coming out of the piezoelectric ceramic round tube after ultrasonic modulation is a Bessel light beam.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the spirit and scope of the invention. The present invention is not to be limited by the specific embodiments disclosed herein, and other embodiments that fall within the scope of the claims of the present application are intended to be within the scope of the present invention.
Claims (6)
1. A large-depth-of-field Bessel beam photoacoustic microscopic imaging device based on ultrasonic modulation comprises: the device comprises a pulse laser (1), a piezoelectric ceramic round tube (6), a three-dimensional scanner (17), a digital pulse delayer (12) and a function generator (10);
the pulse laser (1) is electrically connected with the digital pulse delayer (12) and the workstation (20);
the function generator (10) is electrically connected with the D-type trigger (11) and the piezoelectric ceramic round tube (6);
the workstation (20) is electrically connected with the pulse laser (1), the acquisition card (19) and the three-dimensional scanner (17).
2. A large-depth-of-field Bessel beam photoacoustic microscopic imaging method based on ultrasonic modulation comprises the following steps:
s1: the pulse laser (1) emits laser pulses, the size of a light spot is shaped to be equal to the caliber of the piezoelectric ceramic round tube (6) through the beam expanding system, and then the laser pulses are incident to the piezoelectric ceramic round tube (6) filled with optical transparent liquid;
s2: a function generator (10) is used for applying a sine radio frequency signal to the piezoelectric ceramic circular tube (6), the inner wall and the outer wall of the piezoelectric ceramic circular tube (6) vibrate and generate ultrasonic waves, the ultrasonic waves can periodically modulate the density of local optical transparent liquid, so that the refractive index distribution of the optical transparent liquid is changed, and the refractive index distribution of the optical transparent liquid in the radial direction is represented as zero-order Bessel function distribution and is synchronously changed with the sine radio frequency signal;
s3: the synchronous circuit is used for synchronizing the light emission of the pulse laser (1) and the change of the refractive index of liquid in the piezoelectric ceramic round tube (6), so that the pulse laser (1) emits light on a phase of the change of the refractive index;
the pulse laser (1) emits light when the refractive index of liquid in the piezoelectric ceramic round tube (6) is changed to be positive and maximum by controlling the time delay of the digital pulse time delay device (12), the radial refractive index distribution of the liquid in the piezoelectric ceramic round tube (6) is zero-order Bessel function distribution at the moment, and thus, a light beam which is subjected to ultrasonic modulation and then comes out of the piezoelectric ceramic round tube (6) is a Bessel light beam;
s4, focusing the acquired Bessel light beam on a sample (16) through a fifth condenser lens (9), detecting the generated photoacoustic signal through an acoustic lens, amplifying the photoacoustic signal through an amplifier (18), and finally acquiring the photoacoustic signal by an acquisition card (19);
s5, using the three-dimensional scanner (17) to perform two-dimensional raster scanning to acquire three-dimensional data.
3. The large-depth-of-field Bessel beam photoacoustic microscopy method based on ultrasonic modulation according to claim 2, characterized in that:
the beam expanding system in S1 is composed of a first condenser lens (2) and a second condenser lens (3),
the piezoelectric ceramic round tube (6) is polarized and then is plated on the inner wall and the outer wall to form an inner electrode and an outer electrode, the inner electrode extends from one end to the outer wall, the inner electrode and the outer electrode form two gaps on the outer wall and the end face of the other end,
the laser pulse vertically enters the piezoelectric ceramic round tube (6), in order to avoid asymmetric radial refractive index distribution caused by gravity when the piezoelectric ceramic round tube is horizontally placed, the piezoelectric ceramic round tube (6) is vertically placed, the optical transparent liquid is silicone oil, and the silicone oil has the kinematic viscosity of 50-150cSt, the refractive index of 1-2 and the sound velocity of 800-1200 m/s.
4. The large-depth-of-field Bessel beam photoacoustic microscopy method based on ultrasonic modulation according to claim 2, characterized in that:
s2, the refractive index distribution of the optically transparent liquid after being subjected to ultrasonic modulation is
Wherein n is0Is the static refractive index of the medium, csIs the speed of sound in the medium, ω is the frequency of the drive signal, r and t denote position and time, respectively, J0Is a zero order Bessel function, naIs a constant related to omega, the physical properties of the acoustic medium, and the refractive index distribution behaves as a zero order bessel function and changes synchronously with the drive signal over time.
5. The large-depth-of-field Bessel beam photoacoustic microscopy method based on ultrasonic modulation according to claim 2, characterized in that:
the synchronous circuit of S3 is composed of a three-dimensional scanner (17), a function generator (10), a D-type trigger (11) and a digital pulse delayer (12), wherein the function generator (10) is used for outputting sine radio frequency signals to drive the piezoelectric ceramic round tube (6), the synchronous signals of the sine radio frequency signals are accessed to the clock end of the D-type trigger (11), the three-dimensional scanner (17) outputs position synchronous signals to be accessed to the D end of the D-type trigger (11) every step of horizontal movement, and the position synchronous signals are TTL square wave signals;
according to the characteristic equation of the D-type flip-flop (11), the D-type flip-flop (11) is triggered only when the synchronous signal rises, and the output signal is equal to the input signal; the output end of the D-type trigger (11) is connected with a digital pulse delayer (12), and the output end of the digital pulse delayer (12) is used for triggering the pulse laser (1).
6. The large-depth-of-field Bessel beam photoacoustic microscopy method based on ultrasonic modulation according to claim 2, characterized in that:
s4, the acoustic lens is composed of an ultrasonic transducer (13) and an acoustic-optical coupling prism (14),
the numerical aperture of the acousto-optic coupling prism is 0.5.
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Cited By (2)
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