CN216771488U - Photoacoustic microscopic imaging system for large-depth imaging - Google Patents
Photoacoustic microscopic imaging system for large-depth imaging Download PDFInfo
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- CN216771488U CN216771488U CN202220128530.4U CN202220128530U CN216771488U CN 216771488 U CN216771488 U CN 216771488U CN 202220128530 U CN202220128530 U CN 202220128530U CN 216771488 U CN216771488 U CN 216771488U
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Abstract
The utility model discloses a photoacoustic microscopic imaging system for large-depth imaging, which comprises a signal acquisition device and an imaging processing terminal, wherein the signal acquisition device comprises: the device comprises a pulse laser, a first beam splitter, a first polaroid, a first objective lens, a second beam splitter, a second polaroid, a second objective lens, a helium-neon laser, a polarizer, a dimming slide, a first optical filter, a second optical filter, a prism, a coupling medium, a first analyzer, a second analyzer, a differential detector, a band-pass filter and an amplifier. According to the photoacoustic microscopic imaging system, based on the light beams output by the first light path and the second light path, the light beams are focused on a sample in a high-flux and high-resolution mode through the bidirectional excitation light beams, the phase type total internal reflection sensor is used as a photoacoustic wave detector, the detection of the broadband and high sensitivity of the photoacoustic wave is realized by analyzing the refractive index change of a coupling medium caused by photoacoustic signals, and the imaging quality of the sample is greatly improved.
Description
Technical Field
The utility model relates to the technical field of optical microscopic imaging, in particular to a photoacoustic microscopic imaging system for large-depth imaging.
Background
The photoacoustic microimaging technology is that by means of optical excitation and acoustic detection, the optical absorption characteristic of pigment substances in a tissue sample to pulse laser is utilized to convert optical energy into heat energy, and ultrasonic waves are released due to the instantaneous thermo-elastic effect of the tissue. This ultrasonic signal is detected by an ultrasonic detector, the ultrasonic time of flight of which provides the depth position at which the pigment substance is located; and combining a proper sample scanning mode to obtain a three-dimensional morphological structure image of the sample.
Over ten years, the photoacoustic microscopic imaging technology has shown great application potential in the fields of oncology, brain science and the like. However, there are still many drawbacks to conventional photoacoustic microscopy imaging techniques that need to be overcome. First, due to the strong optical attenuation properties of tissue to laser light, the penetration depth of photoacoustic microscopy imaging techniques is typically around 1 millimeter. This makes it difficult for the photoacoustic microscopy imaging technique to accurately obtain the optical absorption characteristics of deep tissues. Secondly, the traditional piezoelectric ultrasonic transducer widely used for photoacoustic signal detection is limited by physical properties of the transducer, and the detection bandwidth is narrow (generally, only dozens of megahertz), so that the longitudinal resolution of photoacoustic microimaging is limited to dozens of microns, the depth position of an optical absorption substance cannot be accurately reflected, the depth positioning accuracy is influenced, and the three-dimensional image is seriously distorted. In addition, the sensitivity of the piezoelectric type ultrasonic transducer is generally around several hundred pa, which makes the contrast of photoacoustic imaging low, the image quality is poor, and it is difficult to identify a tissue sample of a minute size due to the poor quality of the obtained image. Therefore, the existing photoacoustic microscopic imaging technology has the problem of poor imaging quality.
Disclosure of Invention
The embodiment of the utility model provides a photoacoustic microscopic imaging system for large-depth imaging, aiming at solving the problem of poor imaging quality of the existing photoacoustic microscopic imaging technology.
The embodiment of the utility model provides a photoacoustic microscopic imaging system for large-depth imaging, which comprises a signal acquisition device and an imaging processing terminal, wherein the signal acquisition device comprises: the device comprises a pulse laser, a first beam splitter, a first polaroid, a first objective lens, a second beam splitter, a second polaroid, a second objective lens, a helium-neon laser, a polarizer, a dimming slide, a first optical filter, a second optical filter, a prism, a coupling medium, a first analyzer, a second analyzer, a differential detector, a band-pass filter and an amplifier; the first beam splitter is arranged at the downstream of the pulse laser so as to split the laser output by the pulse laser through the first beam splitter; the first polaroid and the first objective lens are arranged in a first optical path between the first beam splitter and the three-dimensional moving objective table, and the second polaroid and the second objective lens are arranged in a second optical path between the first beam splitter and the three-dimensional moving objective table; the three-dimensional moving object stage is used for placing a sample to be imaged, the coupling medium covers the sample to be imaged, and the prism is arranged on the coupling medium; the light beam output by the first light path and the light beam output by the second light path respectively irradiate the sample to be imaged from the upper side and the lower side so as to obtain a two-dimensional photoacoustic detection signal; the polarizer and the light adjusting slide are arranged in a detection light path between the helium-neon laser and the prism; the detection light beams output by the light adjusting slide are incident to the prism and are totally internally reflected on the bottom surface of the prism; the second beam splitter is arranged at an emergent position of the detection light beam reflected by the prism, so that the emergent detection light is split by the second beam splitter; the first analyzer and the first optical filter are arranged in a first feedback light path between the second beam splitter and the differential detector, and the second analyzer and the second optical filter are arranged in a second feedback light path between the second beam splitter and the differential detector; two detection ports arranged on the differential detector are respectively used for inputting a first detection light beam output by the first optical filter and a second detection light beam output by the second optical filter; the differential detector is electrically connected with the imaging processing terminal through the band-pass filter and the amplifier so as to output a differential detection signal to the imaging processing terminal, and the imaging processing terminal processes the differential detection signal and the two-dimensional photoacoustic detection signal to obtain a display image of the sample to be imaged.
The photoacoustic microscopic imaging system for large-depth imaging is characterized in that the prism is a trapezoidal prism.
The photoacoustic microscopic imaging system for large-depth imaging is characterized in that the refractive index of the prism is larger than 1.3.
The photoacoustic microscopy imaging system for large-depth imaging comprises 1/2 slide glass and 1/4 slide glass.
The photoacoustic microimaging system for large-depth imaging is characterized in that a first confocal lens is arranged between the first polarizer and the first objective lens; and a second confocal lens is arranged between the second polaroid and the second objective lens.
The photoacoustic microscopic imaging system for large-depth imaging is characterized in that a long-focus lens is further arranged between the light adjusting slide and the prism.
The photoacoustic microscopic imaging system for large-depth imaging is characterized in that the coupling medium is normal saline, distilled water or deionized water.
The embodiment of the utility model provides a photoacoustic microscopic imaging system for large-depth imaging, which comprises a signal acquisition device and an imaging processing terminal, wherein the signal acquisition device comprises: the device comprises a pulse laser, a first beam splitter, a first polaroid, a first objective lens, a second beam splitter, a second polaroid, a second objective lens, a helium-neon laser, a polarizer, a dimming slide, a first optical filter, a second optical filter, a prism, a coupling medium, a first analyzer, a second analyzer, a differential detector, a band-pass filter and an amplifier. The photoacoustic microscopic imaging system for large-depth imaging outputs light beams based on the first light path and the second light path, and realizes that the light beams are focused on a sample at high flux and high resolution through the bidirectional excitation light beams, so that the imaging depth of the sample is increased, and the efficient detection of photoacoustic signals is ensured; based on the total internal reflection of the prism, the photoacoustic wave generated by the probe beam after the laser excites the sample is transmitted in the coupling medium and interacts with the evanescent field, thereby causing the phase change of the probe beam. The change of the light intensity of the detection light beam is detected through the differential detector, the high-sensitivity detection of broadband photoacoustic waves is realized, the excitation light can be transmitted to the tissue sample through the scheme with high optical transmittance, the bidirectional excitation of pulse laser can be realized, the photoacoustic imaging depth is increased, and the imaging quality of the sample is greatly improved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
Fig. 1 is a schematic structural diagram of a photoacoustic microscopy imaging system for large-depth imaging provided by an embodiment of the present invention;
FIG. 2 is a schematic flow chart of a photoacoustic microscopy imaging method for large-depth imaging provided by an embodiment of the present invention;
fig. 3 is a schematic diagram of the effect of the large-depth imaging photoacoustic microscopy imaging system provided by the embodiment of the utility model.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It will be understood that the terms "comprises" and/or "comprising," when used in this specification and the appended claims, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.
It is also to be understood that the terminology used in the description of the utility model herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the utility model. As used in the specification of the present invention and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.
It should be further understood that the term "and/or" as used in this specification and the appended claims refers to any and all possible combinations of one or more of the associated listed items and includes such combinations.
In this embodiment, please refer to fig. 1, where fig. 1 is a schematic structural diagram of a photoacoustic microscopy imaging system with large depth imaging according to an embodiment of the present invention. As shown in the figure, the embodiment of the present invention provides a photoacoustic microscopic imaging system for large-depth imaging, where the system includes a signal acquisition device and an imaging processing terminal 20, where the signal acquisition device includes: the device comprises a pulse laser 1, a first beam splitter 2, a first polaroid 11, a first objective 12, a second beam splitter 3, a second polaroid 21, a second objective 22, a He-Ne laser 4, a polarizer 5, a light adjusting slide 6, a first optical filter 31, a second optical filter 41, a prism 7, a coupling medium 8, a first analyzer 32, a second analyzer 42, a differential detector 9, a band-pass filter 18 and an amplifier 19; the first beam splitter 2 is arranged at the downstream of the pulse laser 1, so that the laser output by the pulse laser 1 is split by the first beam splitter 2; the first polarizing plate 11 and the first objective lens 12 are disposed in a first optical path between the first beam splitter 2 and the three-dimensional moving stage 101, and the second polarizing plate 21 and the second objective lens 22 are disposed in a second optical path between the first beam splitter 2 and the three-dimensional moving stage 101. Wherein a first confocal lens 13 is disposed between the first polarizer 11 and the first objective lens 12; a second confocal lens 23 is disposed between the second polarizing plate 21 and the second objective lens 22. The three-dimensional moving object stage 101 is used for placing a sample 10 to be imaged, the coupling medium 8 covers the sample 10 to be imaged, and the prism 7 is arranged on the coupling medium 8; the light beam output by the first optical path and the light beam output by the second optical path respectively irradiate the sample 10 to be imaged from the upper side and the lower side, so as to obtain a two-dimensional photoacoustic detection signal.
The pulse laser 1 may be Nd: YAG (Neodymium-doped yttrium aluminum garnet) pulse laser, wherein the pulse laser can be used for generating photoacoustic exciting light in picosecond level, the first beam splitter 2 is used for splitting the pulse laser output by the pulse laser into two beams, one beam is transmitted by a first optical path, and the other beam is transmitted by a second optical path. The polarizer is used to adjust the polarization direction of the incident beam, the lens is used for focusing or collimating, and the objective lens is used to focus the beam. Pulse laser light in the first optical path is output as linearly polarized light through the first polarizer 11, the linearly polarized light in the first optical path can be expanded through a 4F optical system formed by a pair of confocal lenses, the linearly polarized light in the first optical path changes the light beam direction by the reflector, enters the first objective lens 12 from the upper part to the lower part, passes through the prism 7 and the coupling medium 8, and then is focused and irradiated on the surface of a sample 10 to be imaged. The pulse laser in the second optical path is output as linearly polarized light through the second polarizer 21, the linearly polarized light in the second optical path can be expanded through a 4F optical system formed by a pair of confocal lenses, the linearly polarized light in the second optical path is changed in beam direction by the reflector, enters the second objective lens 22 from the lower direction, and is focused and irradiated on the back of the sample 10 to be imaged. Therefore, the bidirectional excitation light illumination mode is realized, and the sample is simultaneously excited from the upper part and the lower part of the sample to be imaged to generate photoacoustic signals.
The polarizer 5 and the light adjusting slide 6 are arranged in a detection light path between the helium-neon laser 6 and the prism 7; the detection light beam output by the light adjusting slide 6 is incident to the prism 7 and is totally internally reflected at the bottom surface of the prism 7; the second beam splitter 3 is disposed at an exit position where the probe beam is reflected by the prism 7, so as to split the exiting probe beam by the second beam splitter 3; the first analyzer 32 and the first optical filter 31 are arranged in a first feedback optical path between the second beam splitter 3 and the differential detector 9, and the second analyzer 42 and the second optical filter 41 are arranged in a second feedback optical path between the second beam splitter 3 and the differential detector 9; the two detection ports of the differential detector 9 are respectively used for inputting the first detection beam output by the first optical filter 31 and the second detection beam output by the second optical filter 41. Specifically, the light control slide 6 includes 1/2 slides 61 and 1/4 slides 62. Specifically, a telephoto lens 71 is further disposed between the light-adjusting slide 6 and the prism 7. The coupling medium 8 may be normal saline, distilled water or deionized water, and in practical use, distilled water is preferred, and in the most preferred embodiment, deionized water is selected. Specifically, the telephoto lens 71 may be used to slightly focus the probe beam output from the light adjusting slide 6.
More specifically, the prism 7 is a trapezoidal prism. The refractive index of the prism 7 is greater than 1.3, and a Total Internal Reflection (TIR) photoacoustic detector can be constructed based on the prism 7 and the coupling medium 8. In a specific embodiment, the prism 7 may be an isosceles trapezoid prism, so as to improve the symmetry of the light beam propagating in the prism 7, so as to improve the accuracy of subsequent imaging. In addition, the prism can be arranged into prisms with other angles, such as a regular hexagon prism. The refractive index of the prism 7 can be set to be larger than 1.3, a reflecting interface is formed between the prism 7 (optical dense medium) and the coupling medium 8 (optical sparse medium), and when incident light enters the interface at a specific angle, the total internal reflection can be generated when the incident angle is larger than the critical angle. For example, a prism may be made of K9 glass with a refractive index of n-1.51509, and K9 glass (n) may be used to simplify the optical path designK91.51509) a special trapezoidal prism is customized to make the incident beam incident into and emergent from the prism in parallel, and ensure that the incident angle at this time is just the total internal reflection angle, the specific effect is as shown in fig. 3, wherein in fig. 3 ^ A is 114.66 DEG, ^ B is 155.34 DEG, and the upper side length L of the trapezoidal prism is L112.74mm, lower edge length L240mm, total height H1Is 10mm, side height H23.743 mm.
In the embodiment, a phase-type total internal reflection sensing technology is adopted for detecting the photoacoustic signal, the continuous laser is adjusted by the polarizer 5 and the light adjusting glass sheet 6 to obtain elliptically polarized light, the polarized light beam is slightly focused by the long-focus lens 71 and enters the prism 7, and total internal reflection occurs on the bottom surface of the prism 7. After being adjusted by the lens, the reflected light beam is divided into two beams by the second beam splitter 3, and the two beams of light are transmitted to two detection ports of the differential detector through the first feedback light path and the second feedback light path respectively.
The sample is simultaneously excited from the upper part and the lower part of the sample to be imaged by adopting a bidirectional exciting light illumination mode to generate photoacoustic signals, and the photoacoustic signals are detected by using the phase mode total internal reflection optical surface wave sensor, so that the microscopic morphological structure characteristics in a large depth range can be observed by using a photoacoustic microscopic imaging technology under the condition of no mark, and the physiological/pathological information of tissues can be accurately acquired.
The differential detector 9 is electrically connected with the imaging processing terminal 20 through the band-pass filter 18 and the amplifier 19 to output a differential detection signal to the imaging processing terminal 20, and the imaging processing terminal 20 processes the differential detection signal and the two-dimensional photoacoustic detection signal to obtain a display image of the sample to be imaged.
In the photoacoustic imaging process, the sample to be imaged 10 is fixed on the three-dimensional moving stage 101, so that two beams are focused on the sample to be imaged 10 to perform two-dimensional planar scanning to obtain two-dimensional photoacoustic detection signals required for three-dimensional image stacking. The photoacoustic wave generated during scanning is coupled by a coupling medium (aqueous solution), so that the refractive index of the coupling medium is changed, and the photoacoustic signal can be detected by detecting the change.
The technical method in the embodiment has the following characteristics: 1. a bidirectional excitation light illumination mode is established, photoacoustic signals are simultaneously excited from the upper side and the lower side of a biological sample, and the photoacoustic imaging depth is greatly improved. 2. The photoacoustic detection method based on phase mode Total Internal Reflection (TIR) optical surface wave sensing is established, high-sensitivity and broadband photoacoustic signal detection is realized, the depth resolution of a photoacoustic microscopic imaging technology is improved, and the depth position of the microstructure of a sample to be observed is accurately positioned. At the same time, the sensor has optically transparent properties, allowing bi-directional illumination of the photoacoustic excitation beam.
Compared with the traditional photoacoustic microscopic imaging technology, the depth of photoacoustic imaging can be improved by two times in a bidirectional excitation light illumination mode to the maximum extent; the photoacoustic detection method based on the phase optical surface wave sensing improves the detection bandwidth to hundreds of megahertz, increases the detection sensitivity to dozens of pascals, and is obviously superior to a piezoelectric ultrasonic transducer. The two innovative technologies are integrated into a novel photoacoustic microscopic imaging system, and the imaging depth and the longitudinal resolution are improved at the same time, so that high longitudinal resolution and large depth imaging can be performed on biological tissues, and a reliable technical means is provided for observing the three-dimensional morphological structure of a thick tissue sample.
Referring to fig. 2, fig. 2 is a schematic flow chart of a method of a photoacoustic microscopy imaging method for large-depth imaging according to an embodiment of the present invention. The embodiment of the utility model also provides a photoacoustic microscopic imaging method for large-depth imaging, wherein the photoacoustic microscopic imaging method for large-depth imaging is applied to the photoacoustic microscopic imaging system for large-depth imaging, as shown in fig. 2, and the method comprises the steps of S110-S180.
And S110, turning on the pulse laser to output the adjustable pulse laser with specific pulse width and specific wavelength.
Specifically, in the embodiment, the pulse laser may generate an adjustable pulse laser with a specific wavelength of 150 to 400nm, the pulse width of the generated pulse laser is adjustable, the specific pulse width is 50 to 900 picoseconds, for example, for a certain specific tissue sample, the adjustable pulse laser with the specific wavelength of 266nm and the specific pulse width of 800ps (picosecond) may be adopted, the wavelength of the photoacoustic excitation light is not limited to 266nm, and for different samples, the wavelength may be changed, based on the maximum absorption coefficient of the sample and the strongest generated photoacoustic signal.
S120, after the adjustable pulse laser is split by the first beam splitter, one part of the light beam is adjusted to be linearly polarized light beam through the first light path to irradiate the sample to be imaged from the upper part, and the other part of the light beam is adjusted to be linearly polarized light beam through the second light path to irradiate the sample to be imaged from the lower part; the polarization directions of the two beams illuminating the sample to be imaged are different.
The two beams of light which irradiate the sample to be imaged are not necessarily linearly polarized light which is perpendicular to each other, but only need to have different polarization directions, namely two beams of light (elliptical or circularly polarized light) with different vibration directions can be emitted, and the maximum signal and the maximum imaging depth are taken as the criteria. In a preferred embodiment, the two light beams illuminating the sample to be imaged may be arranged with their polarization directions perpendicular to each other.
S130, the two light beams perform two-dimensional plane scanning on the sample to be imaged and detect the change of the coupling medium refractive index caused by the photoacoustic wave generated during scanning, so that a two-dimensional photoacoustic detection signal is obtained.
And S140, starting the helium-neon laser to output continuous laser, adjusting the continuous laser to be a detection beam through the detection light path, and then, enabling the detection beam to enter the prism, wherein the polarized beam is totally internally reflected on the bottom surface of the prism and is split through the second beam splitter.
The wavelength of continuous laser output by the helium-neon laser can be 450-750 nm. If a sample to be imaged is detected, the He-Ne laser can be controlled to output continuous laser with the wavelength of 632.8 nm. The detection light beams output by the light adjusting slide are elliptical polarized light with different phase differences on the vertical component and the horizontal component, the detection light wavelength is not limited to 632.8nm, and different detection light wavelengths correspond to different excitation angles.
S150, a part of the detection beam output by the second beam splitter is input to one detection port of the differential detector through the first feedback optical path, and another part of the detection beam is input to another detection port of the differential detector through the second feedback optical path.
And S160, the differential detector performs differential detection on the light beams from the first feedback light path and the second feedback light path to obtain a differential detection signal, and the differential detection signal is output to the band-pass filter.
And S170, the band-pass filter performs noise filtering on the differential detection signal and then outputs the differential detection signal to the amplifier for amplification to obtain a detection amplification signal.
And S180, the imaging processing terminal acquires the detection amplification signal and the two-dimensional photoacoustic detection signal and carries out three-dimensional image stacking processing to obtain a display image corresponding to the sample to be imaged.
In a specific application process, the imaging processing terminal can be respectively and electrically connected with the pulse laser and the helium-neon laser, so that corresponding parameters are input through the imaging processing terminal to adjust the pulse laser output by the pulse laser and the continuous laser output by the helium-neon laser. In addition, the imaging processing terminal can be electrically connected with the three-dimensional moving object stage, so that corresponding moving parameters are input through the imaging processing terminal to control the precise electric three-dimensional moving object stage to move in three dimensions. The imaging processing terminal can be a desktop computer, a notebook computer, a tablet computer and other terminal equipment which can be used for data and image processing.
The photoacoustic microscopic imaging method for large-depth imaging in the embodiment has the following characteristics: 1. and a bidirectional excitation light path is established, and the imaging depth is increased. The method utilizes the high-efficiency focusing and conduction of the optical element to the excitation beam and the optical transparency of the ultrasonic detector, so that the bidirectional excitation beam can be focused on the tissue sample with high flux and high resolution, the imaging depth of the tissue sample is increased, and the high-efficiency detection of the photoacoustic signal is ensured. 2. Photoacoustic signal detection based on total internal reflection-type phase TIR sensors. After the laser excites the sample, the generated photoacoustic wave is propagated in the coupling medium and interacts with the evanescent field, thereby changing the phase change of the detection light. The change of the polarized light intensity is detected and analyzed by a differential detection method, so that the high-sensitivity detection of the broadband photoacoustic wave is realized. In addition, the sensor has optical transparent characteristic, so that the exciting light is transmitted to the tissue sample with high optical transmittance, and therefore bidirectional excitation of pulse laser can be achieved, and the depth of photoacoustic imaging is increased. The design realizes the large-depth excitation and the high-efficiency detection of the photoacoustic signals of the biological tissues, so that the novel photoacoustic microscopic imaging technology can accurately observe the three-dimensional microscopic morphological structure of the tissue sample.
The embodiment of the utility model provides a photoacoustic microscopic imaging system for large-depth imaging, which comprises a signal acquisition device and an imaging processing terminal, wherein the signal acquisition device comprises: the device comprises a pulse laser, a first beam splitter, a first polaroid, a first objective lens, a second beam splitter, a second polaroid, a second objective lens, a helium-neon laser, a polarizer, a dimming slide, a first optical filter, a second optical filter, a prism, a coupling medium, a first analyzer, a second analyzer, a differential detector, a band-pass filter and an amplifier. The photoacoustic microscopic imaging system for large-depth imaging outputs light beams based on the first light path and the second light path, and realizes that the light beams are focused on a sample at high flux and high resolution by bidirectional excitation light beams, so that the imaging depth of the sample is increased, and the high-efficiency detection of photoacoustic signals is ensured; based on the total internal reflection of the prism, the photoacoustic wave generated by the probe beam after the laser excites the sample is transmitted in the coupling medium and interacts with the evanescent field, thereby causing the phase change of the probe beam. The change of the light intensity of the detection light beam is detected through the differential detector, the high-sensitivity detection of the broadband photoacoustic wave is realized, the excitation light can be transmitted to the tissue sample through high optical transmittance by the scheme, the bidirectional excitation of pulse laser can be realized, the photoacoustic imaging depth is increased, and the imaging quality of the sample is greatly improved.
While the utility model has been described with reference to specific embodiments, the utility model is not limited thereto, and various equivalent modifications and substitutions can be easily made by those skilled in the art within the technical scope of the utility model. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (7)
1. The photoacoustic microscopic imaging system for large-depth imaging is characterized by comprising a signal acquisition device and an imaging processing terminal, wherein the signal acquisition device comprises: the device comprises a pulse laser, a first beam splitter, a first polaroid, a first objective lens, a second beam splitter, a second polaroid, a second objective lens, a helium-neon laser, a polarizer, a dimming slide, a first optical filter, a second optical filter, a prism, a coupling medium, a first analyzer, a second analyzer, a differential detector, a band-pass filter and an amplifier;
the first beam splitter is arranged at the downstream of the pulse laser so as to split the laser output by the pulse laser through the first beam splitter;
the first polaroid and the first objective lens are arranged in a first optical path between the first beam splitter and the three-dimensional moving objective table, and the second polaroid and the second objective lens are arranged in a second optical path between the first beam splitter and the three-dimensional moving objective table;
the three-dimensional moving object stage is used for placing a sample to be imaged, the coupling medium covers the sample to be imaged, and the prism is arranged on the coupling medium; the light beam output by the first light path and the light beam output by the second light path respectively irradiate the sample to be imaged from the upper side and the lower side so as to obtain a two-dimensional photoacoustic detection signal;
the polarizer and the light adjusting slide are arranged in a detection light path between the helium-neon laser and the prism; the detection light beam output by the light adjusting slide enters the prism and is totally internally reflected on the bottom surface of the prism;
the second beam splitter is arranged at an emergent position of the detection light beam reflected by the prism so as to split the emergent detection light beam by the second beam splitter;
the first analyzer and the first optical filter are arranged in a first feedback light path between the second beam splitter and the differential detector, and the second analyzer and the second optical filter are arranged in a second feedback light path between the second beam splitter and the differential detector; two detection ports arranged on the differential detector are respectively used for inputting a first detection light beam output by the first optical filter and a second detection light beam output by the second optical filter;
the differential detector is electrically connected with the imaging processing terminal through the band-pass filter and the amplifier so as to output a differential detection signal to the imaging processing terminal, and the imaging processing terminal processes the differential detection signal and the two-dimensional photoacoustic detection signal to obtain a display image of the sample to be imaged.
2. The large depth imaging photoacoustic microscopy imaging system of claim 1, wherein the prism is a trapezoidal prism.
3. A large depth imaging photoacoustic microscopy imaging system as set forth in claim 1 or 2, wherein the refractive index of the prism is greater than 1.3.
4. The large-depth imaging photoacoustic microscopy imaging system of claim 3, wherein the light-modulating slides comprise 1/2 slides and 1/4 slides.
5. The large-depth imaging photoacoustic microscopy imaging system of claim 3, wherein a first confocal lens is disposed between the first polarizer and the first objective lens; and a second confocal lens is arranged between the second polaroid and the second objective lens.
6. The large-depth imaging photoacoustic microscopy imaging system of claim 3, wherein a tele lens is further disposed between the light modulating slide and the prism.
7. The large-depth imaging photoacoustic microscopy imaging system of claim 3, wherein the coupling medium is physiological saline, distilled water, or deionized water.
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