CN113670827A - Polarization sensitive optical coherence-hyperspectral microimaging device and detection method thereof - Google Patents
Polarization sensitive optical coherence-hyperspectral microimaging device and detection method thereof Download PDFInfo
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/21—Polarisation-affecting properties
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- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
- G01N21/45—Refractivity; Phase-affecting properties, e.g. optical path length using interferometric methods; using Schlieren methods
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/21—Polarisation-affecting properties
- G01N2021/216—Polarisation-affecting properties using circular polarised light
Abstract
The invention discloses a polarization-sensitive optical coherence-hyperspectral microimaging device and an imaging method thereof, wherein the device comprises a laser light source component, an imaging detection component, a signal acquisition/processing component and a sample stage, wherein the laser light source component is used for providing laser in a linear polarization state to irradiate tissues, and the imaging detection component is used for carrying out systematic imaging on a sample to be detected; the laser light source assembly, the imaging detection assembly and the signal acquisition/processing assembly are electrically connected in sequence, the laser light source assembly is connected with the signal acquisition/processing assembly, and the imaging detection assembly is connected with the sample stage. The invention realizes that a plurality of complementary parameters of the sample can be obtained by one-time imaging by using the polarized light source and reasonably combining two imaging technologies, gets rid of the limitation that only partial information can be obtained by single imaging, and saves the detection time and the economic cost; the polarization characteristic of light is utilized, the imaging quality of the traditional imaging mode is improved, and the contrast of an image is improved.
Description
Technical Field
The invention belongs to the technical field of biomedical imaging, and particularly relates to a polarization-sensitive optical coherence-hyperspectral microimaging device and a detection method thereof.
Background
Optical Coherence Tomography (OCT) is a biomedical imaging technique based on the principle of optical low-coherence interference to achieve high sensitivity, high resolution, and high imaging speed on three-dimensional spatial information inside a sample. Compared with the conventional imaging technology, OCT has gained wide attention as a novel imaging mode since its first introduction by university of labor in ma province in 1991. Through the development of recent decades, the OCT makes great progress in the aspects of detection sensitivity, imaging speed and the like, and also promotes the development of functional imaging technologies such as label-free OCT blood flow motion radiography and the like. At present, OCT is widely used for clinical diagnosis and treatment of ophthalmology and cardiovascular department, and has important scientific value and application prospect in the medical fields of brain science, tumor, digestive tract, respiratory tract, skin, development and the like.
The ordinary OCT images a tissue in two or three dimensions using the scattering property of a biological tissue, and reflects structural information of the tissue. Since the advent of OCT, some functional OCT imaging techniques have been introduced, such as polarization-sensitive OCT (PS-OCT) used herein, which uses the vector characteristics of light to detect birefringence information of biological tissues, measurement of birefringence characteristics of joint ligaments by polarization OCT, study of the influence of the tension or relaxation state of ligaments on birefringence characteristics, quantitative determination of skin burn level, and the like.
High spectral microscopy (HSI) utilizes specific absorption of different components in the tissue, and the absorption process of photons depends on energy level transition of molecules, so that the absorption spectrum of a specific wave band can be used as a basis for identifying specific molecules, and the microenvironment of the tissue can be detected non-invasively by utilizing the change of the optical properties of the tissue. Therefore, the hyperspectral microimaging is used as a novel medical image form, has the advantages of non-contact, no damage, high precision, good repeatability and the like, is widely applied to the field of biological detection, and has great potential in the aspects of noninvasive disease diagnosis and surgical guidance.
Disclosure of Invention
The invention mainly aims to overcome the defects of the prior art and provide a polarization-sensitive optical coherence-hyperspectral microscopic imaging device and a detection method thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a polarization-sensitive optical coherence-hyperspectral microimaging device which comprises a laser light source component, an imaging detection component, a signal acquisition/processing component and a sample stage, wherein the laser light source component is used for providing laser in a linear polarization state to irradiate tissues, and the imaging detection component is used for performing system imaging on a sample to be detected; the laser light source assembly, the imaging detection assembly and the signal acquisition/processing assembly are electrically connected in sequence, the laser light source assembly is connected with the signal acquisition/processing assembly, and the imaging detection assembly is connected with the sample stage;
the laser light source assembly comprises a super-continuum spectrum laser light source, a linear polaroid, a beam expanding collimator and a2 x 2 optical fiber coupler, the super-continuum spectrum laser light source, the linear polaroid, the beam expanding collimator and the 2 x 2 optical fiber coupler are sequentially connected, and the 2 x 2 optical fiber coupler is respectively connected with the imaging detection assembly, the signal acquisition/processing assembly and the sample stage;
the imaging detection assembly comprises an adjustable diaphragm, a quarter-wave plate A, a reflector, a quarter-wave plate B, a two-dimensional scanning galvanometer and a flat field objective lens; the adjustable diaphragm, the quarter-wave plate A and the reflector are sequentially connected to form a reference arm; the quarter-wave plate B, the two-dimensional scanning galvanometer and the flat field objective are sequentially connected to form a sample arm; the flat field objective is connected with the sample stage, and the adjustable diaphragm and the quarter-wave plate B are both connected with the laser light source component;
the signal acquisition/processing assembly comprises a collimator, a polarization beam splitter, a spectrometer A, a spectrometer B, a coaxial cable, an acquisition card and a computer, wherein the collimator and the polarization beam splitter are sequentially connected and then simultaneously connected with the spectrometer A and the spectrometer B, the collimator is connected with a 2X 2 optical fiber coupler of a laser light source assembly, the acquisition card is connected with the computer, the computer is connected with a two-dimensional scanning vibrating mirror driver, and the acquisition card is respectively connected with the spectrometer A and the spectrometer B through the coaxial cable.
Preferably, a quarter-wave plate a is arranged between the adjustable diaphragm and the reflector, the quarter-wave plate a is 22.5 degrees relative to the linearly polarized incident light, the quarter-wave plate changes the polarization state of the incident light by 22.5 degrees, and the incident light passes through the quarter-wave plate twice after being reflected by the reflector, so that the linear polarization state of the light is changed by 45 degrees relative to the incident light when the light exits from the reference arm; a quarter wave plate B is arranged between the 2 multiplied by 2 optical fiber coupler and the two-dimensional scanning galvanometer, the quarter wave plate B is 45 degrees relative to the linear polarization incident light, and the quarter wave plate B converts the linear polarization incident light into circular polarization light and then enters the sample stage.
Preferably, a linear polarizer is arranged between the supercontinuum laser and the beam expanding collimator, so that light emitted by the supercontinuum laser is linearly polarized light.
Preferably, the laser light source, the reference arm reflected light, the sample arm scattered light and the 2 × 2 fiber coupler are strictly optically coaxial; the flat field objective, the collimator, the polarization beam splitter, the spectrometer A and the spectrometer B are strictly coaxial in optics.
Preferably, the polarizing beam splitter obtains data in both horizontal and vertical polarization directions simultaneously in one scan.
Preferably, the laser light source is divided into two beams of light by a2 × 2 fiber coupler; the reference arm reflected light and the sample arm scattered light are combined into a beam of light through a 2X 2 optical fiber coupler.
Preferably, the computer is provided with an acquisition control and signal processing system;
the acquisition control and signal processing system adopts an acquisition control and signal processing system written by LabView and MATLAB, and continuously changes the input voltage of the vibrating mirror through the program control of the LabView, so that the deflection angle of the vibrating mirror is changed, incident light is focused on different positions of the surface of a sample, and the local scanning of the surface of the sample is realized; and then converting the acquired light intensity signal into an electric signal, and then obtaining an image which is the same as that of the sample in the scanning area by using an MATLAB program so as to realize the functions of scanning and imaging the sample.
Preferably, the combination of optical coherence and hyperspectral is not a simple combination, but rather, two systems share the same laser, optical path, and the same set of signal acquisition device.
The invention also provides a detection method of the polarization-sensitive optical coherence-hyperspectral microimaging device, which comprises the following steps:
(1) placing an imaging detection assembly directly above the surface of the sample;
(2) laser emitted by the supercontinuum laser irradiates the surface of a sample through a linear polarizer, a beam expanding collimator, a2 multiplied by 2 fiber coupler, a quarter wave plate B, a two-dimensional scanning galvanometer and a flat field objective in sequence, so that incident light is focused on the surface of the sample;
(3) emergent light irradiates a sample, the light is absorbed, scattered and reflected on the surface or shallow surface of the sample, the back scattered light and the reflected light of the sample are received through a signal acquisition/processing assembly, and the light intensity change is a required signal;
(4) the back scattered light and the reflected light which carry the structure and the function information of the sample after passing through the sample are irradiated on the spectrometer A and the spectrometer B through the flat field objective lens, the two-dimensional scanning galvanometer, the quarter wave plate B, the 2 multiplied by 2 optical fiber coupler, the collimator and the polarization beam splitter in sequence, and the change of the light intensity on the spectrometer A and the spectrometer B is the required signal in the horizontal and vertical polarization directions; changing the deflection angles of the shafts of the two-dimensional scanning galvanometers X, Y to deflect incident light, wherein the acquisition card acquires data once when the two-dimensional scanning galvanometers deflect once;
(5) after the complete signal is collected, a two-dimensional image and a three-dimensional image of the tissue sample are reconstructed by a maximum value projection method, so that the structural and functional imaging of the sample is realized.
Preferably, the pulse laser wavelength of the laser light source is 400-2400nm, the pulse width is 100ps, and the repetition frequency is continuously adjustable within the range of 0.1-25 MHz.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the invention considers the respective advantages and disadvantages of optical coherence tomography and hyperspectral microscopy, and then organically combines the two imaging modes by using the polarization characteristic of light to realize the complementary imaging effect, and compared with a single imaging mode, the invention can obtain more abundant and accurate information of a sample in a multi-dimension way by using the vector characteristic of light, namely the combination of the polarization effect on the two technologies:
with respect to a single common hyperspectral imaging, although it is possible to obtain the spectral information differences of different characteristic tissues, studies have shown that in the course of disease detection, due to the wrong statement of melanin and blood content, the use of a single hyperspectral imaging usually results in the wrong estimation of deoxyhemoglobin, oxyhemoglobin and total hemoglobin, which is particularly important for assessing the pathological changes of skin cancer, since the high concentration of melanin interferes with the quantitative algorithm for determining blood and collagen distribution. In addition, the hyperspectral imaging cannot obtain accurate imaging of organisms with tissues with different scattering properties, including structural information in the depth direction of the sample;
if a biological sample is imaged using only polarized light, we can obtain birefringence information of the sample and polarization components in two orthogonal directions, and calculate an image based on the degree of linear polarization, but according to existing research work, this method is only effective for benign pigmented nevus portions with high melanin concentration;
the single OCT can perform structural imaging by detecting interference signals of different fault planes of the biological tissue by using the light scattering property of the biological tissue, but it is difficult to obtain a more comprehensive biological internal structure reflected by the biological tissue through optical absorption difference imaging, and molecular information of the biological tissue is lost;
based on the above limitations, we combine two imaging methods based on the use of polarized light, and can solve the following problems: 1. at high melanin content, the erroneous estimation of the oxyhemoglobin and deoxyhemoglobin contents is corrected by using the polarization characteristics of light, and the superficial melanin can be accurately separated, so that the distribution of oxyhemoglobin and deoxyhemoglobin can be accurately evaluated; 2. specific absorption spectrum (hyperspectral), structure distribution (OCT) and birefringence information (polarization characteristic of light) of a sample are obtained; 3. the contrast of the image is improved; 4. the inconvenience that only partial information can be extracted in a single imaging mode and the detection needs to be carried out for multiple times is avoided; 5. the combination of the polarization characteristics of the light realizes the complementation of the two imaging modes.
(2) The invention combines two imaging modes not simply, but leads the two to share a supercontinuum light source, a light path and the same set of detection device, and the hyperspectral imaging only needs to shield a reference arm of the OCT system, thus realizing the imaging function of the bimodal system. By the method, the complexity of the system is greatly reduced, the cost is reduced, the system building is easier to integrate, and compared with a simple combination, the method is more economical and convenient, and the imaging quality is more stable.
(3) The invention selects the supercontinuum laser, the output wavelength can be continuously adjusted within the range of 400-2400nm, the output wavelength can be continuously adjusted, and the spectral information of samples under different wave bands and the fine structure change at different depth positions are detected; meanwhile, compared with the commonly used bandwidth of 50-100nm of the OCT system, the system has wider bandwidth of the selected light source, so that the axial resolution of the OCT system is improved more obviously. The method has a great promoting effect on the realization of deeper, finer and clearer imaging targets.
(4) The invention utilizes a polarization beam splitter to simultaneously obtain two orthogonal polarization components, namely horizontal polarization component and vertical polarization component, in one scanning, thereby avoiding the inconvenience that two polarization components are obtained only by rotating the angle of a polaroid in two scans in the traditional technology.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, 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 only some embodiments of the present application, 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 polarization-sensitive optical coherence-hyperspectral microimaging apparatus according to embodiment 1 of the invention;
FIG. 2 is a schematic diagram of a spectrometer according to an embodiment of the present invention.
The reference numbers illustrate:
1 is a grating, 2 is a focusing lens, 3 is a CCD, 1-1 is a supercontinuum laser, 1-2 is a linear polaroid, 1-3 is a beam expanding collimator, 1-4 is a2 x 2 optical fiber coupler, 2-1 is an adjustable diaphragm, 2-2 is a quarter wave plate A, 2-3 is a reflector, 2-4 is a quarter wave plate B, 2-5 is a two-dimensional scanning galvanometer, 2-6 is a flat field objective lens, 3-1 is a sample stage, 4-1 is a collimator, 4-2 is a polarization beam splitter, 4-3 is a spectrometer A, 4-4 is a spectrometer B, 4-5 is an acquisition card, and 4-6 is a computer.
Detailed Description
In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. It is to be understood that the embodiments described are only a few embodiments of the present application and not all embodiments. 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 application.
Reference in the specification to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the specification. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. It is explicitly and implicitly understood by one skilled in the art that the embodiments described herein can be combined with other embodiments.
Example 1
As shown in fig. 1, the present embodiment provides a polarization-sensitive optical coherence-hyperspectral microimaging apparatus, which includes a laser light source assembly, an imaging detection assembly, a signal acquisition/processing assembly, and a sample stage 3-1, where the laser light source assembly, the imaging detection assembly, and the signal acquisition/processing assembly are sequentially connected, the laser light source assembly is electrically connected with the signal acquisition/processing assembly, and the signal detection assembly is connected with the sample stage 3-1; the device not only uses linearly polarized light to image OCT and realizes polarization sensitive OCT (PS-OCT), but also the incident light for hyperspectral microimaging is linearly polarized light. Compared with the traditional light source, the light source provided by the invention utilizes the polarization characteristic of light, and solves the problem of error estimation of oxyhemoglobin and deoxyhemoglobin of biological tissues under high melanin content. Compared with the traditional OCT technology, the method can only obtain the structural distribution of the sample, and can obtain the birefringence information of the sample on the original basis by using polarized light imaging, thereby being convenient for more comprehensively and deeply knowing the change of the internal composition of the sample and simultaneously improving the contrast of the image.
Further, the laser light source assembly comprises a supercontinuum laser 1-1, a linear polaroid 1-2, a beam expanding collimator 1-3 and a2 x 2 optical fiber coupler 1-4, wherein the supercontinuum laser 1-1, the linear polaroid 1-2, the beam expanding collimator 1-3 and the 2 x 2 optical fiber coupler 1-4 are sequentially connected, and the 2 x 2 optical fiber coupler 1-4 is respectively connected with the imaging detection assembly and the signal acquisition/processing assembly.
Furthermore, the imaging detection assembly comprises an adjustable diaphragm 2-1, a quarter wave plate A2-2, a reflector 2-3, a quarter wave plate B2-4, a two-dimensional scanning galvanometer 2-5 and a flat field objective lens 2-6; the adjustable diaphragm 2-1, the quarter-wave plate A2-2 and the reflector 2-3 are sequentially connected to form a reference arm; the quarter-wave plate B2-4, the two-dimensional scanning galvanometer 2-5 and the flat field objective lens 2-6 are sequentially connected to form a sample arm; the flat field objective 2-6 is connected with the sample stage 3-1, and the adjustable diaphragm 2-1 and the quarter-wave plate B2-4 are connected with the laser light source component.
Furthermore, a quarter-wave plate A is arranged between the adjustable diaphragm and the reflector, the quarter-wave plate A is 22.5 degrees relative to the linearly polarized incident light, the quarter-wave plate mainly has the function of changing the polarization state of the incident light by 22.5 degrees and penetrates through the quarter-wave plate twice after being reflected by the reflector, so that the linear polarization state of the light is changed by 45 degrees relative to the incident light when the light is emitted from the reference arm; a quarter wave plate B is arranged between the 2 multiplied by 2 optical fiber coupler and the two-dimensional scanning galvanometer, the quarter wave plate B is 45 degrees relative to the linear polarization incident light, and the quarter wave plate B converts the linear polarization incident light into circular polarization light and then enters the sample stage.
It can be understood that the imaging detection component mainly has the main function that when laser in the reference arm irradiates a certain point on a sample, light can be scattered in the sample or specifically absorbed by the sample, and scattered light at different depth positions of the sample interferes with reflected light of the reference arm under certain conditions, so that interference signals at different depth positions of the sample can be extracted by utilizing the signal acquisition/processing component; then, by utilizing the specific absorption of the sample to light, the absorption spectra of different components in the sample and the absorption spectra of the sample under different wavelengths can be obtained; and the information of each point on the sample can be obtained through the deflection movement of the two-dimensional scanning galvanometer, so that the two-dimensional or three-dimensional imaging is carried out on the sample.
Further, the signal acquisition/processing assembly comprises a collimator 4-1, a polarization beam splitter 4-2, a spectrometer A4-3 and a spectrometer B4-4, the device comprises a coaxial cable, an acquisition card 4-5 and a computer 4-6, wherein the collimator 4-1 and a polarization beam splitter 4-2 are sequentially connected and then are simultaneously connected with a spectrometer A4-3 and a spectrometer B4-4, the collimator 4-1 is connected with a2 x 2 optical fiber coupler 1-4 of a laser light source component, the acquisition card 4-5 is connected with the computer 4-6, the computer 4-6 is connected with a two-dimensional scanning vibrating mirror 2-5 driver, and the acquisition card 4-5 is respectively connected with the spectrometer A4-3 and the spectrometer B4-4 through the coaxial cable.
It will be appreciated that this embodiment eliminates the use of a single polarizer in the signal acquisition/processing assembly, and uses a polarizing beamsplitter so that data in both the horizontal and vertical polarization directions can be acquired simultaneously in a single scan.
Furthermore, the computer 4-6 is provided with an acquisition control and signal processing system; the acquisition control and signal processing system adopts an acquisition control and signal processing system which is automatically compiled by LabView and MATLAB; continuously changing the input voltage of the galvanometer through program control of LabView to change the deflection angle of the galvanometer, so that the incident light is focused on different positions of the surface of the sample, thereby realizing local scanning of the surface of the sample; and then converting the acquired light intensity signal into an electric signal, and then obtaining an image which is the same as that of the sample in the scanning area by using an MATLAB program so as to realize the functions of scanning and imaging the sample.
Further, the laser light source is divided into two beams of light by a2 x 2 optical fiber coupler; the reference arm reflected light and the sample arm scattered light are combined into a beam of light through a 2X 2 optical fiber coupler.
Furthermore, the laser light source, the reflected light of the reference arm, the scattered light of the sample arm and the 2 x 2 optical fiber coupler are strictly in optical coaxial; the flat field objective, the collimator, the polarization beam splitter, the spectrometer A and the spectrometer B are strictly coaxial in optics.
In the embodiment, the pulse laser generated by the supercontinuum laser 1-1 is focused on a sample through the flat field objective lens 2-6, and light does not propagate along a straight line in the tissue, but is absorbed, scattered and reflected; the light carrying the OCT signal is obtained by detecting the back scattering light radiated from the surface or the shallow surface of the sample; then receiving the light reflected back from the surface of the sample to obtain hyperspectral signal light carrying spectral information; then, the driving voltage of the two-dimensional scanning galvanometer is changed so as to change the deflection angle of the X axis and the Y axis of the galvanometer, so that incident light is deflected, and the acquisition card acquires data once when the two-dimensional scanning galvanometer deflects once; the light irradiates different positions of the sample to obtain changed light intensity, the light intensity is received by the spectrometer, the spectrometer converts the light signal into an electric signal, and the computer finishes signal acquisition. And after all the signals are collected, reconstructing a photoacoustic two-dimensional image and a photoacoustic three-dimensional image of the tissue sample by a maximum value projection method.
Example 2
As shown in fig. 2, the detection method using the polarization-sensitive optical coherence hyperspectral microscopy imaging apparatus of example 1 includes the following steps:
(1) the back scattered light and the reflected light carrying the sample information are irradiated on the grating 1 through an imaging detection component, a2 multiplied by 2 optical fiber coupler, a collimator and a polarization beam splitter, and the grating 1 enables the amplitude or the phase (or both) of the incident light to be subjected to periodic spatial modulation through a regular structure;
(2) incident light is separated into a spectrum with a certain width through the grating 1 and is focused into a narrow line through the focusing lens 2;
(3) the CCD 3 is used for receiving monochromatic light corresponding to different wavelengths, and each pixel on the CCD 3 receives the monochromatic light corresponding to different wavelengths, so that imaging of high spectrum and different depths of OCT is realized;
the pulse laser wavelength range of the supercontinuum laser source is 550-1100nm, and the repetition frequency is 5 MHz.
The method for establishing the three-dimensional image preferably adopts the following method: and (3) taking the same time scale for all signals and projecting longitudinal sections, reconstructing a three-dimensional image on three-dimensional reconstruction software VolView3.2 by using the image obtained after projection, and rotating the whole three-dimensional image in the three-dimensional reconstruction software to obtain a three-dimensional image with any view angle.
The polarization-sensitive optical coherence-hyperspectral microimaging device can be applied to the field of biomedicine, and particularly can be applied to research of morphological structures, physiological characteristics and pathological characteristics of biological tissues.
It should be noted that, for the sake of simplicity, the foregoing method embodiments are described as a series of acts or combinations, but those skilled in the art should understand that the present invention is not limited by the described order of acts, as some steps may be performed in other orders or simultaneously according to the present invention.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, but should be considered as the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A polarization-sensitive optical coherence-hyperspectral microimaging device is characterized by comprising a laser light source assembly, an imaging detection assembly, a signal acquisition/processing assembly and a sample stage, wherein the laser light source assembly is used for providing laser in a linear polarization state to irradiate tissues, and the imaging detection assembly is used for performing system imaging on a sample to be detected; the laser light source assembly, the imaging detection assembly and the signal acquisition/processing assembly are electrically connected in sequence, the laser light source assembly is connected with the signal acquisition/processing assembly, and the imaging detection assembly is connected with the sample stage;
the laser light source assembly comprises a super-continuum spectrum laser light source, a linear polaroid, a beam expanding collimator and a2 x 2 optical fiber coupler, the super-continuum spectrum laser light source, the linear polaroid, the beam expanding collimator and the 2 x 2 optical fiber coupler are sequentially connected, and the 2 x 2 optical fiber coupler is respectively connected with the imaging detection assembly, the signal acquisition/processing assembly and the sample stage;
the imaging detection assembly comprises an adjustable diaphragm, a quarter-wave plate A, a reflector, a quarter-wave plate B, a two-dimensional scanning galvanometer and a flat field objective lens; the adjustable diaphragm, the quarter-wave plate A and the reflector are sequentially connected to form a reference arm; the quarter-wave plate B, the two-dimensional scanning galvanometer and the flat field objective are sequentially connected to form a sample arm; the flat field objective is connected with the sample stage, and the adjustable diaphragm and the quarter-wave plate B are both connected with the laser light source component;
the signal acquisition/processing assembly comprises a collimator, a polarization beam splitter, a spectrometer A, a spectrometer B, a coaxial cable, an acquisition card and a computer, wherein the collimator and the polarization beam splitter are sequentially connected and then simultaneously connected with the spectrometer A and the spectrometer B, the collimator is connected with a 2X 2 optical fiber coupler of a laser light source assembly, the acquisition card is connected with the computer, the computer is connected with a two-dimensional scanning vibrating mirror driver, and the acquisition card is respectively connected with the spectrometer A and the spectrometer B through the coaxial cable.
2. The polarization-sensitive optical coherence-hyperspectral microimaging apparatus according to claim 1, wherein a quarter-wave plate a is disposed between the adjustable diaphragm and the mirror, the quarter-wave plate a is 22.5 ° with respect to the linearly polarized incident light, the quarter-wave plate changes the polarization state of the incident light by 22.5 °, and the light passes through the quarter-wave plate twice after being reflected by the mirror, so that the linear polarization state of the light changes by 45 ° with respect to the incident light when exiting from the reference arm; a quarter wave plate B is arranged between the 2 multiplied by 2 optical fiber coupler and the two-dimensional scanning galvanometer, the quarter wave plate B is 45 degrees relative to the linear polarization incident light, and the quarter wave plate B converts the linear polarization incident light into circular polarization light and then enters the sample stage.
3. The polarization-sensitive optical coherence hyperspectral microimaging apparatus according to claim 1, wherein a linear polarizer is disposed between the supercontinuum laser and the beam-expanding collimator, so that the light emitted from the supercontinuum laser is linearly polarized.
4. The polarization-sensitive optical coherence-hyperspectral microimaging device according to claim 1, wherein the laser light source, the reference arm reflected light, the sample arm scattered light and the 2 x 2 fiber coupler are strictly optically coaxial; the flat field objective, the collimator, the polarization beam splitter, the spectrometer A and the spectrometer B are strictly coaxial in optics.
5. The polarization-sensitive optical coherence-hyperspectral microscopic imaging apparatus according to claim 1, wherein the polarizing beam splitter obtains data in both horizontal and vertical polarization directions simultaneously in one scan.
6. The polarization-sensitive optical coherence-hyperspectral microscopic imaging device according to claim 1, wherein the laser light source is divided into two beams of light by a2 x 2 fiber coupler; the reference arm reflected light and the sample arm scattered light are combined into a beam of light through a 2X 2 optical fiber coupler.
7. The polarization-sensitive optical coherence-hyperspectral microscopic imaging device according to claim 1, wherein the computer is provided with an acquisition control and signal processing system;
the acquisition control and signal processing system adopts an acquisition control and signal processing system written by LabView and MATLAB, and continuously changes the input voltage of the vibrating mirror through the program control of the LabView, so that the deflection angle of the vibrating mirror is changed, incident light is focused on different positions of the surface of a sample, and the local scanning of the surface of the sample is realized; and then converting the acquired light intensity signal into an electric signal, and then obtaining an image which is the same as that of the sample in the scanning area by using an MATLAB program so as to realize the functions of scanning and imaging the sample.
8. The polarization-sensitive optical coherence-hyperspectral microimaging apparatus of claim 1, wherein the combination of optical coherence and hyperspectral is not a simple combination, but rather two systems share the same laser, optical path and the same set of signal acquisition devices.
9. The detection method of the polarization-sensitive optical coherence-hyperspectral microscopic imaging device according to any one of claims 1 to 8, characterized by comprising the following steps:
(1) placing an imaging detection assembly directly above the surface of the sample;
(2) laser emitted by the supercontinuum laser irradiates the surface of a sample through a linear polarizer, a beam expanding collimator, a2 multiplied by 2 fiber coupler, a quarter wave plate B, a two-dimensional scanning galvanometer and a flat field objective in sequence, so that incident light is focused on the surface of the sample;
(3) emergent light irradiates a sample, the light is absorbed, scattered and reflected on the surface or shallow surface of the sample, the back scattered light and the reflected light of the sample are received through a signal acquisition/processing assembly, and the light intensity change is a required signal;
(4) the back scattered light and the reflected light which carry the structure and the function information of the sample after passing through the sample are irradiated on the spectrometer A and the spectrometer B through the flat field objective lens, the two-dimensional scanning galvanometer, the quarter wave plate B, the 2 multiplied by 2 optical fiber coupler, the collimator and the polarization beam splitter in sequence, and the change of the light intensity on the spectrometer A and the spectrometer B is the required signal in the horizontal and vertical polarization directions; changing the deflection angles of the shafts of the two-dimensional scanning galvanometers X, Y to deflect incident light, wherein the acquisition card acquires data once when the two-dimensional scanning galvanometers deflect once;
(5) after the complete signal is collected, a two-dimensional image and a three-dimensional image of the tissue sample are reconstructed by a maximum value projection method, so that the structural and functional imaging of the sample is realized.
10. The detecting method as claimed in claim 8, wherein the laser light source has a pulse laser wavelength of 400-2400nm, a pulse width of 100ps, and a repetition frequency of 0.1-25 MHz.
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