CN114264614A - Label-free photoacoustic pathological microscopic imaging system and imaging method - Google Patents
Label-free photoacoustic pathological microscopic imaging system and imaging method Download PDFInfo
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Abstract
The invention discloses a label-free photoacoustic pathological microscopic imaging system and method. The system comprises: the optical surface wave sensor is used for bearing the tissue sample and the water layer; an excitation light generating device; a detection light generating device; an objective lens, which makes the excitation light beam and the detection light beam incident on the tissue sample through the optical surface wave sensor; processing and control means for controlling the movement of the excitation beam to scan the tissue sample; the optical surface wave sensor is used for totally reflecting the detection light beam and emitting the reflected detection light beam through the objective lens; and the processing and control device is used for carrying out beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beam to obtain a target image. The imaging system is high in integration and integration degree, an optical surface wave sensor is used as a glass slide for bearing a tissue sample, high-frequency signals can be reserved to a great extent, longitudinal resolution is improved, and in addition, an objective lens is shared by an excitation light beam and a detection light beam, so that coaxiality of two light beams can be better guaranteed.
Description
Technical Field
The invention relates to the field of microscopic imaging, in particular to a label-free photoacoustic pathological microscopic imaging system and method.
Background
At present, the photoacoustic microscopy imaging technology is a new biological imaging mode which develops the fastest in the last decade, and has great potential in the aspect of pathological detection, and a Liuwen faithful team of the university of northwest of the United states utilizes the photoacoustic imaging technology to detect the eye structure of a mouse, so that the retinal blood oxygen saturation and melanin concentration can be better detected compared with other technologies, and the clinical transfer of photoacoustic imaging is further promoted. The U.S. Catherine Martel group combined a photoacoustic microscope with a micro-electro-mechanical system scanning mirror for use in the pathological examination of lymphatic vessels. Although the photoacoustic microscopic imaging technology has been developed, due to the restriction of the properties of piezoelectric materials, the defects of narrow detection bandwidth and low sensitivity of photoacoustic systems are common. The narrow detection bandwidth leads to low longitudinal resolution of photoacoustic imaging, and further causes inaccurate depth direction positioning, which is not beneficial to actual pathological imaging. In order to solve this problem, the sensing technology based on the optical surface wave is applied to the field of photoacoustic imaging, and high sensitivity of the photoacoustic imaging system is realized by detecting the phase change of the detection optical phase.
Compared with the traditional photoacoustic sensor, the optical surface wave sensor has the advantages of high sensitivity and broadband photoacoustic detection; in a novel field of photoacoustic imaging based on optical surface wave sensing, a representative Virginia university in the United states establishes a set of photoacoustic microscopy system by using a plasma detection technology, and high spatial resolution is realized compared with the traditional photoacoustic microscopy imaging. The Yanfan and the like of Shenzhen university have made great progress in the in-situ phase type TIR ultrasonic sensing technology, the transverse resolution of the image obtained by the built system can reach 5.8 mu m, and the system has higher sensitivity and wider detection bandwidth compared with the traditional photoacoustic imaging system.
However, although the existing phase-type TIR ultrasonic sensing system can image tissue pathology well, this technique still has defects, such as the tissue sample is far away from the surface acoustic wave sensor, and the photoacoustic high-frequency signal emitted by the sample needs to propagate in water for a long distance, as shown in fig. 1, the tissue sample a and the water layer b are placed on a glass slide c, the glass slide c is placed on a two-dimensional displacement platform d, a prism e and an objective lens f are arranged above the two-dimensional displacement platform d, after the excitation beam is focused by the objective lens f, the excitation beam is incident to the prism e and is incident to the tissue sample a, the detection beam emitted by the TIR detector g is emitted to the surface of the water layer b through the prism e and is totally reflected, in the case that the surface of the water layer b is close to the bottom surface of the prism e (i.e. the distance between the prism e and the water layer b is reduced to the greatest extent, and the movement of the two-dimensional displacement platform d is not interfered), according to the measurement data, the difference between the lower surface of the prism e and the tissue sample a is 5mm, and the difference of the 5mm distance has a great influence on high-frequency information, namely, the acoustic wave bandwidth of the prism is reduced from 1GHz to 170MHz, so that the longitudinal resolution of an image is directly reduced, and the three-dimensional imaging capability of the system is influenced.
Disclosure of Invention
The invention aims to provide a label-free photoacoustic pathology microscopic imaging system and a label-free photoacoustic pathology microscopic imaging method, and aims to solve the problems that the longitudinal resolution of an image generated by an existing phase-type TIR ultrasonic sensing system is low, and the existing probe beam and the existing coaxial are poor.
In order to solve the technical problems, the invention aims to realize the following technical scheme: there is provided a label-free photoacoustic pathology microscopy imaging system, comprising:
an optical surface wave sensor for carrying a tissue sample and a water layer, wherein the tissue sample is immersed in the water layer;
excitation light generating means for generating an excitation light beam;
a probe light generating device for generating a probe light beam;
the objective lens is used for focusing the excitation light beam and converting the detection light beam into a parallel detection light beam, so that the excitation light beam and the detection light beam are incident on the tissue sample through the surface optical wave sensor;
processing and control means for controlling the movement of the excitation beam to scan the tissue sample;
the optical surface wave sensor is also used for totally reflecting the detection light beam to enable the reflected detection light beam to be emitted through the objective lens;
and the processing and control device is used for receiving the returned detection light beams, and performing beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
In addition, an object of the present invention is to provide an imaging method of a label-free photoacoustic pathology microscopic imaging system, which adopts the label-free photoacoustic pathology microscopic imaging system as described above, and includes:
placing a tissue sample and an aqueous layer on the optical surface wave sensor, wherein the tissue sample is immersed in the aqueous layer;
the excitation light generating device outputs and generates an excitation light beam, and the detection light generating device outputs and generates a detection light beam;
the objective lens focuses the excitation light beam and converts the detection light beam into parallel detection light beams, so that the excitation light beam and the detection light beam are incident on a tissue sample through the surface acoustic wave sensor;
the processing and control device controls the excitation beam movement to scan the tissue sample;
the optical surface wave sensor totally reflects the detection light beam to enable the reflected detection light beam to be emitted through the objective lens;
and the processing and control device receives the returned detection light beams, and performs beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
The embodiment of the invention discloses a label-free photoacoustic pathological microscopic imaging system and an imaging method, wherein the imaging system comprises: an optical surface wave sensor for carrying a tissue sample and a water layer, wherein the tissue sample is immersed in the water layer; excitation light generating means for generating an excitation light beam; a probe light generating device for generating a probe light beam; the objective lens is used for focusing the excitation light beam and converting the detection light beam into a parallel detection light beam, so that the excitation light beam and the detection light beam are incident on the tissue sample through the surface optical wave sensor; processing and control means for controlling the movement of the excitation beam to scan the tissue sample; the optical surface wave sensor is also used for totally reflecting the detection light beam to enable the reflected detection light beam to be emitted through the objective lens; and the processing and control device is used for receiving the returned detection light beams, and performing beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
Compared with the prior art, the imaging system has higher integration degree and integration degree, simplifies the complexity of the imaging system and is easier to apply to the field of biomedical imaging; in the second aspect, the system uses the optical surface wave sensor as a glass slide for bearing a tissue sample, so that the photoacoustic high-frequency signal emitted by the tissue sample does not need to be remotely transmitted in water, the high-frequency signal can be greatly reserved, the longitudinal resolution is further improved, and high-sensitivity and broadband photoacoustic detection is realized; in the third aspect, the imaging system of the application enables the excitation light beam and the detection light beam to share one objective lens, so that the coaxiality of the two light beams can be better ensured, and the imaging transverse resolution is further 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 prior art phase-type TIR ultrasonic sensing system;
FIG. 2 is a schematic structural diagram of a label-free photoacoustic pathology microscopy imaging system provided by an embodiment of the present invention;
fig. 3 is a schematic flow chart of an imaging method of the label-free photoacoustic pathology microscopy imaging system according to the embodiment of the present invention.
Description of reference numerals:
a. a tissue sample; b. a water layer; c. a glass slide; d. a two-dimensional displacement platform; e. a prism; f. an objective lens; g. a TIR detector;
1. an optical surface wave sensor; 2. an excitation light generating device; 3. a detection light generating device; 4. an objective lens; 5. a processing terminal; 6. an optical galvanometer; 7. a fifth mirror; 8. a first lens; 9. a beam splitting device; 10. a first reflector; 11. a second reflector; 12. a first analyzer; 13. a second analyzer; 14. a third reflector; 15. a fourth mirror; 16. a differential detector; 17. a second lens; 18. a sixth mirror; 19. a polarizing plate; 20. 1/2 a wave plate; 21. 1/4 wave plate.
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 invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. 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 and includes any and all possible combinations of one or more of the associated listed items.
Referring to fig. 2, fig. 2 is a schematic structural diagram of a label-free photoacoustic pathology microscopy imaging system according to an embodiment of the present invention;
the embodiment of the invention provides a label-free photoacoustic pathology microscopic imaging system, which comprises:
an optical surface wave sensor 1 for carrying a tissue sample and a water layer, wherein the tissue sample is immersed in the water layer;
excitation light generating means 2 for generating an excitation light beam;
a probe light generating device 3 for generating a probe light beam;
an objective lens 4 for focusing the excitation light beam and for converting the probe light beam into a parallel probe light beam, so that the excitation light beam and the probe light beam are incident on the tissue sample through the surface acoustic wave sensor 1;
processing and control means for controlling the movement of the excitation beam to scan the tissue sample;
the optical surface wave sensor 1 is further configured to totally reflect the probe beam, so that the reflected probe beam is emitted through the objective lens 4;
and the processing and control device is used for receiving the returned detection light beams, and performing beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
In this embodiment, a tissue sample (tissue or cell) and a water layer are directly placed on an optical surface wave sensor 1, an excitation light generating device 2 outputs an excitation light beam, a detection light generating device 3 outputs a detection light beam, an objective lens 4 focuses the excitation light beam, and the objective lens 4 converts the detection light beam into a parallel light beam to be emitted, under the control of a processing and control device, the emitted excitation light beam and the emitted detection light beam are emitted to the tissue sample, wherein the excitation light beam and the detection light beam are transmitted through the optical surface wave sensor 1 to be incident to the tissue sample, and the excitation light beam is driven to move to scan the tissue sample through the control of the processing and control device, in this embodiment, the detection light beam is reflected to the objective lens 4 through the total reflection of the optical surface wave sensor 1, the processing and control device receives the detection light beam emitted after passing through the objective lens 4, and carrying out beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beam to obtain a target image, wherein the optical surface wave sensor 1 is used for carrying out photoacoustic induction on the total internal reflection retraced light beam.
The application utilizes the optical surface wave sensor 1 to replace the function of a glass slide in the prior art, so that the optical surface wave sensor 1 can realize the function of bearing the tissue sample, the distance between the optical surface wave sensor 1 and the tissue sample can be greatly reduced, the photoacoustic high-frequency signal sent by the tissue sample does not need to be remotely transmitted in water, the high-frequency signal can be greatly reserved, and the longitudinal resolution is improved.
It should be noted that, as shown in fig. 1 in the prior art, the lower surface of the prism e is 5mm away from the tissue sample a, which results in the mismatch between the refractive index of water and the refractive index of the prism e, i.e. aberration, and the light intensity of the focal point is decreased, and the lateral resolution of the image is decreased, in the present application, since the tissue sample is directly placed on the surface acoustic wave sensor 1, the distance between the two is extremely small, and the adverse effect caused by the mismatch between the refractive indexes of water can be ignored, meanwhile, since the positions where the excitation beam and the detection beam converge are different in the prior art, the perfect coaxiality cannot be achieved, and the imaging quality is also affected, the present application can better ensure the coaxiality of the two beams by focusing the excitation beam and the detection beam on the same objective 4, i.e. since the two beams pass through the same objective 4, so as to improve the lateral resolution of the imaging, the photoacoustic signal detection with high sensitivity is also ensured, and the accurate imaging of histopathology and even cytopathology can be realized.
In addition, the microscope imaging system of the application simplifies the complexity of the system while realizing the advantages of no mark, high resolution and high sensitivity, so that the system tends to be more integrated.
In a specific embodiment, the objective lens 4 is located below the surface acoustic wave sensor 1, and the optical galvanometer 6 is located below the objective lens 4.
Through the design, the complexity of the system can be simplified, and each structure of the system is more compact.
In a specific embodiment, the excitation light generator 2 is a laser with a wavelength of 266nm, or 532nm, or 560nm, and the detection light generator 3 is a helium-neon laser with a wavelength of 632.8 nm.
In this embodiment, 560nm excitation light is commonly used for hemoglobin imaging, 532nm excitation light is commonly used for eyeball melanin imaging, and 266nm excitation light is the optimal wavelength for cell nucleus imaging, that is, the wavelength can be changed for different samples, based on the maximum absorption coefficient of the tissue sample and the strongest generated photoacoustic signal; similarly, the wavelength of the probe beam is not limited to 632.8nm, and different probe wavelengths correspond to different excitation angles, so that the detailed description is omitted herein.
In a specific embodiment, the processing and controlling device includes a processing terminal 5 and a plurality of optical galvanometers 6 in signal connection with the processing terminal 5, the processing terminal 5 is configured to control the optical galvanometers 6 to move, wherein the optical galvanometers 6 are located between the excitation light generating device 2 and the objective lens 4.
In this embodiment, the processing terminal 5 includes but is not limited to a computer, and the movement of the optical galvanometer 6 can be controlled by inputting a movement command on the processing terminal 5, wherein the two optical galvanometers 6 are provided, and compared with the phenomenon that the two-dimensional moving platform in the prior art has a slow movement speed, the two optical galvanometers 6 and the processing terminal 5 are used in cooperation, so that the tissue sample can be rapidly and stably scanned by the probe beam. Compared with the existing photoacoustic imaging system which mostly uses a precise displacement control platform to move a tissue sample, the photoacoustic imaging system can overcome the problems that the scanning time is long and the imaging speed is slow due to the fact that the moving speed of a two-dimensional displacement platform is slow by controlling the movement of an optical galvanometer 6 through a processing terminal 5.
In a specific embodiment, a plurality of reflectors and lenses are sequentially disposed between the optical galvanometer 6 and the excitation light generator 2 along a light path direction, wherein the lenses are configured to expand the excitation light beams.
In this embodiment, the number of the reflectors, that is, the fifth reflector 7 shown in fig. 2, is 1, and the fifth reflector 7 is used for reflecting a light beam and changing a propagation direction of the light beam, and the number of the lenses, that is, the first lens 8 shown in fig. 2, is two, and the first lens 8 is used for expanding an excitation light beam and ensuring that the excitation light beam is filled in the optical galvanometer 6.
In a specific embodiment, a plurality of reflectors, polarizers 19 and wave plates are sequentially disposed between the detection light generating device 3 and the objective lens 4 along the optical path direction.
In this embodiment, the mirror is the sixth mirror 18 shown in fig. 2, the sixth mirror 18 is used for reflecting the light beam and changing the propagation direction of the light beam, the polarizer 19 is used for increasing the linear polarization ratio of the detection light beam, and the wave plate is used for adjusting the polarization direction of the linearly polarized light.
In a specific embodiment, the reflecting mirror and the polarizer are provided with one, the wave plate is provided with two, and the two wave plates include 1/2 wave plates 20 and 1/4 wave plates 21 which are sequentially arranged along the optical path direction.
In this embodiment, the 1/2 wave plate 20 is used to continuously adjust the polarization direction of linearly polarized light, and the 1/4 wave plate 21 is used to modulate the linearly polarized light into elliptically polarized light, where it should be noted that the initial probe beam is not limited to linearly polarized light, but may also be elliptically or circularly polarized light, with the highest sensitivity of the signal being taken as the criterion.
In a specific embodiment, the processing and control apparatus further includes a beam splitting device 9, a first mirror 10, a second mirror 11, a first analyzer 12, a second analyzer 13, a third mirror 14, a fourth mirror 15, and a differential detector 16, where the beam splitting device 9 is located between the objective lens 4 and the terminal, and the beam splitting device 9 is configured to split the folded probe beam to obtain a first split beam and a second split beam;
the first reflector 10, the first analyzer 12 and the second reflector 11 are sequentially located in the optical path direction of the first beam splitter;
the third reflector 14, the second analyzer 13 and the fourth reflector 15 are sequentially located in the optical path direction of the second beam splitter;
the differential detector 16 is configured to receive the first split beam and the second split beam, and perform photoacoustic signal synthesis;
the processing terminal 5 is in signal connection with the differential detector 16, and is further configured to receive the synthesized photoacoustic signal and perform data analysis and image reconstruction.
In this embodiment, the beam splitting device 9 is a beam splitter for splitting the probe beam into two parts, and the device for splitting the mixed beam into s-polarized light and p-polarized light is not only a Polarization Beam Splitter (PBS) but also a polarization beam splitter such as a wollaston prism.
The first mirror 10, the second mirror 11, the third mirror 14 and the fourth mirror 15 are used for reflecting light beams and changing the propagation direction of the light beams, the first analyzer 12 and the second analyzer 13 are used for enabling the deflection direction of the polarized light to propagate along the short axis direction or the long axis direction, and the differential detector 16 is used for detecting photoacoustic signals.
In a specific embodiment, a plurality of lenses are disposed between the objective lens 4 and the beam splitting device 9.
In the present embodiment, the lens is the second lens 17 shown in fig. 2, and the second lens 17 is provided with one lens and is used for focusing the probe beam of total reflection.
In this embodiment, 266nm excitation light in the microscopic imaging system is emitted by the excitation light generating device 2, the excitation light beam is expanded by the 4f system formed by a pair of confocal lenses after propagating forward through the reflector 7, it is ensured that the excitation light beam can fill the optical galvanometer 6, the optical galvanometer 6 is controlled by a computer, the expanded excitation light beam can scan on the tissue sample rapidly and stably, the excitation light beam is focused on the tissue sample through the objective lens 4, the tissue sample can emit photoacoustic pulse waves under the action of the excitation light beam, the photoacoustic pulse waves can cause refractive index changes of water, the photoacoustic waves propagate to the surface (upper) of the optical surface wave sensor 1 in the coupling medium water and interact with the evanescent field, wherein the evanescent field is generated when the detection light beam performs total internal reflection.
For the detection part, a 632.8nm laser (3) is used as a detection light source in the system, emitted linearly polarized light (detection light beam) is reflected by a reflector, the polarization degree of the linearly polarized light is better through a polarizing plate, the polarized light passes through an 1/2 wave plate and a 1/4 wave plate so as to be modulated into elliptically polarized light with different phase differences in the s and p directions, the elliptically polarized light passes through an objective lens 4 so as to obtain parallel light and irradiate the light on an optical surface wave sensor 1 for total internal reflection, the photoacoustic pulse wave obtained by excitation of excitation light changes the refractive index of water and changes the phase difference of the reflected s and p polarized light, namely, the excitation light beam acts on a tissue sample, the tissue sample emits ultrasonic information, and the detection light beam receives the information (namely, the phase change) when passing through the interface of the optical surface wave sensor 1 and the water, finally, the required image information is obtained by detecting the phase change.
The specific receiving process is as follows: reflected light (detection light beams) is focused by the second lens 17 and then passes through the beam splitter, the detection light beams are divided into two light beams, the two light beams are respectively transmitted to the two analyzers (12, 13), the deflection directions of the two light beams are respectively along the short axis direction and the long axis direction of the elliptical polarization light beams, the efficiency of converting the phase difference of the s light beam and the p light beam caused by the sound wave in the short axis direction into light intensity is high, the two light beams are mainly used for sensing, the conversion efficiency in the long axis direction is low, and the two light beams are used as reference arms. After deflection by the reflectors (10, 11, 14, 15), two beams of detection light enter the two detection units of the differential detector 16 in parallel, that is, the detection of the photoacoustic signal is finally realized by the differential detector 16, and the photoacoustic signal is stored in a computer for subsequent data analysis and image reconstruction.
In the microscope imaging system that is provided by this application, optical surface wave sensor 1 is by ingenious utilization, both played the effect of a year thing, can become optical surface wave sensor 1 again and transmit optical surface wave, make pathological tissue and detecting beam's distance closer, detecting beam can be better acquire high frequency signal to further improve the longitudinal resolution who forms images of system, and showing and having simplified system complexity, make the system tend to the integration, be applied to biomedical research field more easily.
The excitation light beam and the detection light beam are transmitted by the objective lens 4 at the same time, the coaxiality of two beams of light can be better guaranteed, the transverse resolution of imaging can be improved, high-sensitivity photoacoustic signal detection is guaranteed, the system is miniaturized more, and the integration level is improved.
Compared with the traditional photoacoustic imaging system, the microscopic imaging system has the advantages of being wide in detection bandwidth and high in system sensitivity, and the photoacoustic imaging does not need fluorescent labeling, so that better support is provided for 'label-free, high-resolution and high-sensitivity' observation of a tissue sample.
As shown in fig. 3, the present invention also provides an imaging method of the label-free photoacoustic pathology microscopic imaging system, which uses the label-free photoacoustic pathology microscopic imaging system as described above, and comprises the following steps:
s101, placing a tissue sample and a water layer on the optical surface wave sensor 1, wherein the tissue sample is immersed in the water layer;
s102, the excitation light generating device 2 outputs and generates an excitation light beam, and the detection light generating device 3 outputs and generates a detection light beam;
s103, the objective lens 4 focuses the excitation light beam and converts the detection light beam into a parallel detection light beam, so that the excitation light beam and the detection light beam are incident on the tissue sample through the surface acoustic wave sensor 1;
s104, the processing and control device controls the excitation light beam to move so as to scan the tissue sample;
s105, the optical surface wave sensor 1 totally reflects the probe beam, so that the reflected probe beam is emitted through the objective lens 4;
and S106, the processing and control device receives the returned detection light beam, and performs beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beam to obtain a target image.
In the method, the optical surface wave sensor 1 is skillfully utilized, so that the optical surface wave sensor 1 can play a role of carrying objects and can transmit the optical surface waves, the distance between pathological tissues and the detection light beam is closer, namely the detection light beam can better acquire a high-frequency signal, and the longitudinal resolution of imaging is further improved.
The excitation light beam and the detection light beam are transmitted by the objective lens 4 at the same time, the coaxiality of the two beams of light can be better guaranteed, the transverse resolution of imaging can be improved, high-sensitivity photoacoustic signal detection is guaranteed, and meanwhile the system formed by the method is enabled to be more miniaturized, and the integration level is improved.
Compared with the imaging method of the traditional photoacoustic imaging system, the imaging method of the microscopic imaging system has the characteristics of wider detection bandwidth and higher system sensitivity, and the characteristic that the photoacoustic imaging does not need fluorescent labels, and provides better support for the observation of 'no labels, high resolution and high sensitivity' of tissue samples.
It can be clearly understood by those skilled in the art that, for convenience and brevity of description, the specific working process of the method described above may refer to the corresponding process in the foregoing system embodiment, and is not described herein again.
While the invention has been described with reference to specific embodiments, the invention 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 invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. A label-free photoacoustic pathology microscopy imaging system, comprising:
an optical surface wave sensor for carrying a tissue sample and a water layer, wherein the tissue sample is immersed in the water layer;
excitation light generating means for generating an excitation light beam;
a probe light generating device for generating a probe light beam;
the objective lens is used for focusing the excitation light beam and converting the detection light beam into a parallel detection light beam, so that the excitation light beam and the detection light beam are incident on the tissue sample through the surface optical wave sensor;
processing and control means for controlling the movement of the excitation beam to scan the tissue sample;
the optical surface wave sensor is also used for totally reflecting the detection light beam to enable the reflected detection light beam to be emitted through the objective lens;
and the processing and control device is used for receiving the returned detection light beams, and performing beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
2. The unlabeled photoacoustic pathology microscopy imaging system according to claim 1, wherein said processing and control means comprises a processing terminal, a plurality of optical galvanometers in signal connection with said processing terminal, said processing terminal being adapted to control the movement of said optical galvanometers, wherein said optical galvanometers are located between said excitation light generating means and said objective lens.
3. The system of claim 2, wherein a plurality of reflectors and lenses are sequentially disposed between the optical galvanometer and the excitation light generator along the optical path, wherein the lenses are configured to expand the excitation light beam.
4. The label-free photoacoustic pathology microscopy imaging system according to claim 2, wherein said processing and control means further comprises beam splitting means, a first mirror, a second mirror, a first analyzer, a second analyzer, a third mirror, a fourth mirror, a differential detector, said beam splitting means being located between said objective lens and the terminal, said beam splitting means being configured to split the folded probe beam into a first split beam and a second split beam;
the first reflector, the first analyzer and the second reflector are sequentially positioned in the optical path direction of the first beam splitter;
the third reflector, the second analyzer and the fourth reflector are sequentially positioned in the direction of the light path of the second sub-beam;
the differential detector is used for receiving the first beam splitting and the second beam splitting and performing photoacoustic signal synthesis;
and the processing terminal is in signal connection with the differential detector and is also used for receiving the synthesized photoacoustic signal and performing data analysis and image reconstruction.
5. The label-free photoacoustic pathology microscopy imaging system of claim 4, wherein a number of lenses are disposed between the objective lens and the beam splitting device.
6. The label-free photoacoustic pathology microscopy imaging system of claim 2, wherein said objective lens is located below said optical surface wave sensor and said optical galvanometer is located below said objective lens.
7. The label-free photoacoustic pathology microscopy imaging system of claim 1, wherein the excitation light generating device is a 266nm or 532nm or 560nm wavelength laser and the probe light generating device is a 632.8nm wavelength helium-neon laser.
8. The label-free photoacoustic pathology microscopy imaging system according to claim 1, wherein a plurality of reflectors, polarizers and wave plates are sequentially disposed between the probe light generating device and the objective lens along the optical path direction.
9. The markerless photoacoustic pathology microscopy imaging system of claim 8, wherein the mirror and polarizer are provided in one, the wave plate is provided in two, and the two wave plates comprise 1/2 wave plate and 1/4 wave plate sequentially arranged along the optical path direction.
10. A label-free photoacoustic pathology microscopic imaging method using the label-free photoacoustic pathology microscopic imaging system according to any one of claims 1 to 9, comprising:
placing a tissue sample and an aqueous layer on the optical surface wave sensor, wherein the tissue sample is immersed in the aqueous layer;
the excitation light generating device outputs and generates an excitation light beam, and the detection light generating device outputs and generates a detection light beam;
the objective lens focuses the excitation light beam and converts the detection light beam into parallel detection light beams, so that the excitation light beam and the detection light beam are incident on a tissue sample through the surface acoustic wave sensor;
the processing and control device controls the excitation beam movement to scan the tissue sample;
the optical surface wave sensor totally reflects the detection light beam to enable the reflected detection light beam to be emitted through the objective lens;
and the processing and control device receives the returned detection light beams, and performs beam splitting, photoacoustic signal synthesis and image reconstruction processing on the detection light beams to obtain a target image.
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Cited By (2)
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|---|---|---|---|---|
| CN116499975A (en) * | 2023-06-29 | 2023-07-28 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
| CN117030621A (en) * | 2023-09-11 | 2023-11-10 | 深圳大学 | Marking-free laser histological imaging system and imaging method |
-
2022
- 2022-01-25 CN CN202210088842.1A patent/CN114264614A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116499975A (en) * | 2023-06-29 | 2023-07-28 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
| CN116499975B (en) * | 2023-06-29 | 2023-09-22 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
| CN117030621A (en) * | 2023-09-11 | 2023-11-10 | 深圳大学 | Marking-free laser histological imaging system and imaging method |
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