CN104237134A - Associated chromatographic method and device - Google Patents
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Abstract
一种关联层析方法和装置，该方法包括下列步骤：①固定样品；②激光器发出一束激光照射到样品上，超声波发射器发出的超声也作用到样品上；③从热光源或赝热光源出射的一束物光经过位相共轭镜反射后照射到样品上，经样品内传播后再出射，出射光用光强探测器探测，获得第一组信号光；④从热光源或者赝热源发出的另一束参考光由空间分辨能力的探测器探测获得第二组信号光；或者根据已知的赝热源统计分布计算获得第二组信号光；⑤两组信号进行关联运算给出关联层析图像。本发明可以得到超声焦点处的关联成像。关联层析可以动态无损地获取生物软组织内部光学分辨率的光学成像。
A correlation chromatography method and device, the method comprising the following steps: ①fixing the sample; ②a laser emits a beam of laser light to irradiate the sample, and the ultrasound emitted by the ultrasonic transmitter also acts on the sample; ③from a thermal light source or a pseudothermal light source The outgoing beam of object light is reflected by the phase conjugate mirror and then irradiates the sample, and then exits after propagating in the sample. The outgoing light is detected by a light intensity detector to obtain the first set of signal light; ④ emitted from a thermal light source or a pseudo-thermal source The other beam of reference light is detected by a detector with spatial resolution to obtain the second group of signal light; or the second group of signal light is obtained by calculation according to the known statistical distribution of pseudoheat sources; image. The present invention can obtain associated imaging at the ultrasonic focal point. Correlation tomography can dynamically and nondestructively acquire optical imaging with optical resolution inside biological soft tissues.
Description
技术领域technical field
本发明涉及光学成像，特别是一种关联层析方法和装置。The invention relates to optical imaging, in particular to a correlation tomography method and device.
背景技术Background technique
时间反演超声辅助光学聚焦方法首次实现了动态、无损地汇聚光到生物软组织内。用该方法进行生物组织内的光学层析也受到了广泛的关注，一些文章和专利披露了相关的研究工作进展。在此我们以专利US8450674B2“Acoustic assistedphase conjugate optical tomography(超声辅助位相共轭光学层析)”为例简要分析一下现有技术方案存在的问题。The time-reversal ultrasound-assisted optical focusing method realizes the dynamic and non-destructive focusing of light into biological soft tissue for the first time. Optical tomography in biological tissue using this method has also received extensive attention, and some articles and patents have disclosed the progress of related research work. Here we take the patent US8450674B2 "Acoustic assisted phase conjugate optical tomography" as an example to briefly analyze the problems existing in the existing technical solutions.
专利US8450674B2披露了一种对内部埋藏有一个或多个荧光媒介的样品进行层析的光学显微镜方法和系统，该系统的特征在于，包含一个被测样品和一台激光器，激光器发出的激光照射到样品上；一个超声源，超声源向样品发射超声并在其内部形成一个超声焦点，在此焦点处，超声会调制穿过的光的频率，从而产生一个频率调制过的光束；一个数字位相共轭镜，可以反射从激光器发出的参考光束而产生频率调制光波的位相共轭波前；还有一个探测器，用于探测共轭波前汇聚到样品内而激发的荧光信号进行成像。该发明的基本结构和原理如图1所示。入射激光401(例如，Crystal Laser Rubicon,波长532nm,脉宽20ns,重复频率1kHz，又或者是SpectraPhysics Navigator,波长范围532-533nm,当波长选择532nm时,脉宽7ns,重复频率20kHz)照射到样品1上。样品是由两层生物组织模体102夹着荧光目标物体101(例如，一种荧光燃料，激发/发射峰是532/554nm)组成。同时，脉冲超声(例如，焦斑宽100微米)发射到样品1上，调节荧光物质到超声焦点201处。脉冲转换器204(例如，Olympus V313-SM,15MHz,或者Olympus V3330,50MHz)由函数发生器202和信号放大器203驱动，PC计算机控制激光脉冲和超声脉冲的同步以及数字式位相共轭镜3对波前的记录和反演。位相共轭波前聚焦到超声焦点而激发出的荧光405从样品出射，出射光经过滤波收集到达探测器5，给出采集信号。其中，数字式位相共轭镜的基本结构如图2所示。Patent US8450674B2 discloses an optical microscope method and system for performing chromatography on samples embedded with one or more fluorescent media. The system is characterized in that it includes a sample to be tested and a laser, and the laser emitted by the laser is irradiated to On the sample; an ultrasonic source, the ultrasonic source emits ultrasonic waves to the sample and forms an ultrasonic focal point inside, at this focal point, the ultrasonic wave will modulate the frequency of the passing light, thereby producing a frequency modulated beam; a digital phase common The conjugate mirror can reflect the reference beam emitted from the laser to generate the phase conjugate wavefront of the frequency modulated light wave; there is also a detector, which is used to detect the fluorescent signal excited by the conjugate wavefront converging into the sample for imaging. The basic structure and principle of the invention are shown in Figure 1. Incident laser 401 (for example, Crystal Laser Rubicon, wavelength 532nm, pulse width 20ns, repetition frequency 1kHz, or SpectraPhysics Navigator, wavelength range 532-533nm, when the wavelength is 532nm, pulse width 7ns, repetition frequency 20kHz) irradiates the sample 1 on. The sample is composed of two layers of biological tissue phantom 102 sandwiching a fluorescent target object 101 (for example, a fluorescent dye with excitation/emission peaks at 532/554 nm). At the same time, pulsed ultrasound (for example, focal spot width 100 μm) is emitted onto the sample 1 to adjust the fluorescent substance to the ultrasound focus 201 . Pulse converter 204 (for example, Olympus V313-SM, 15MHz, or Olympus V3330, 50MHz) is driven by function generator 202 and signal amplifier 203, PC computer controls the synchronization of laser pulse and ultrasonic pulse and digital phase conjugate mirror 3 pairs Recording and inversion of wavefronts. The fluorescence 405 excited by focusing the phase conjugate wavefront to the ultrasonic focus exits from the sample, and the exiting light is filtered and collected to reach the detector 5 to give an acquisition signal. Among them, the basic structure of the digital phase conjugate mirror is shown in Figure 2.
从样品出射的散射波前403和从激光器出射光束分离出来的一束参考光束406在CCD上形成稳定的干涉图案，读取干涉图案并计算提取散射波前。空间光调制器(spatial light modulator，简称为SLM)与电荷耦合元件相机(charge-coupleddevice，简称为CCD)之间满足严格的镜像对称，调节SLM的像素使之和计算得到的CCD上的波前分布同样满足镜像对称。空白光照射SLM，反射波前404就是光学位相共轭输出。The scattered wavefront 403 emitted from the sample and a reference beam 406 separated from the emitted beam of the laser form a stable interference pattern on the CCD, and the interference pattern is read to calculate and extract the scattered wavefront. Strict mirror symmetry is satisfied between the spatial light modulator (spatial light modulator, referred to as SLM) and the charge-coupled device camera (charge-coupled device, referred to as CCD), and the pixels of the SLM are adjusted to match the calculated wavefront on the CCD The distribution also satisfies mirror symmetry. Blank light illuminates the SLM, and the reflected wavefront 404 is the optical phase conjugate output.
这一方法可以突破散射限制，在生物软组织的弥散区域内进行光学成像，并且成像深度动态可调，无损伤，可以实现生物软组织内的荧光成像。存在的问题是成像的分辨率是超声分辨率。This method can break through the scattering limit and perform optical imaging in the diffuse area of biological soft tissue, and the imaging depth is dynamically adjustable without damage, and can realize fluorescence imaging in biological soft tissue. The problem is that the resolution of the imaging is ultrasound resolution.
专利US8450674B2的核心技术是时间反演超声调制光学聚焦((Honglin Liu*,Xiao Xu*)and Lihong V.Wang.Time-reversed ultrasonically encoded optical focusinginto turbid media.Nature Photonics,5,154-157(2011).)，即利用超声调制散射光的频率，在超声的焦点位置创造一个频率调制光的虚拟源，在样品外记录该虚拟源所发出的散射光的波前，位相共轭散射波前回到虚拟源位置实现了光在散射体内的聚焦。专利US8450674B2把这一技术应用到对软组织内的荧光团物质进行成像。显然，时间反演超声调制光学聚焦技术对虚拟源和位相共轭镜之间的频率调制光的散射过程可以追溯而使之透明化。但从虚拟源再出射时，光又将经历一个随机散射过程，丢失所携带的位置信息，从而失去定位功能，只能给出荧光的强度信息，且分辨率由超声焦点决定。必须扫描超声焦点记录对应的荧光强度，扫描强度的变化中反映了结构和/或成分的变化。The core technology of patent US8450674B2 is time-reversed ultrasonic modulation optical focusing ((Honglin Liu*, Xiao Xu*) and Lihong V.Wang. Time-reversed ultrasonically encoded optical focusing into turbid media. Nature Photonics,5,154-157(2011).) , that is, use ultrasound to modulate the frequency of the scattered light, create a virtual source of frequency-modulated light at the focus of the ultrasound, record the wavefront of the scattered light emitted by the virtual source outside the sample, and the phase conjugate scattered wavefront returns to the virtual source position Focusing of light within the scattering volume is achieved. Patent US8450674B2 applies this technology to the imaging of fluorophore substances in soft tissues. Apparently, the time-reversal ultrasound-modulated optical focusing technique can trace back the scattering process of the frequency-modulated light between the virtual source and the phase-conjugate mirror to make it transparent. However, when re-exiting from the virtual source, the light will undergo a random scattering process, lose the position information it carries, and thus lose the positioning function. Only the intensity information of the fluorescence can be given, and the resolution is determined by the ultrasonic focus. Structural and/or compositional changes must be reflected in changes in the ultrasound focus that must be scanned to record the corresponding fluorescence intensities.
在经典热光源后者赝热光源的关联成像装置中，入射的(赝)热光源被分为两束，物体放置在其中一束物光路，用没有空间分辨能力的一个点或者面探测器收集经过物体衍射的光，另一束光参考光照射在有空间分辨能力的探测器，例如CCD上，对两组信号做关联运算可以得到物体的相干成像。已经证明关联成像具有散射抑制作用(Wenlin Gong and Shensheng Han.Correlated imaging in scattering media,OpticsLetters,36,394(2011).)，即，经物体衍射后的光在到达探测器前所经历的散射不会明显地降低关联成像的成像质量。但光在到达目标区域前所经历的散射过程会削弱物光路和参考光之间的关联性，从而降低成像质量。换言之，关联成像抑制了后续散射，等效透明了后续散射介质。In the correlative imaging device of classical thermal light source and pseudothermal light source, the incident (pseudo) thermal light source is divided into two beams, and the object is placed in one of the object optical paths, and collected by a point or surface detector without spatial resolution. After the light diffracted by the object, another beam of reference light is irradiated on a detector with spatial resolution, such as a CCD, and the coherent imaging of the object can be obtained by performing correlation operations on the two sets of signals. Correlated imaging has been proved to have a scattering suppression effect (Wenlin Gong and Shensheng Han. Correlated imaging in scattering media, Optics Letters, 36, 394 (2011).), that is, the scattering experienced by the light diffracted by the object before reaching the detector will not be obvious reduce the imaging quality of associated imaging. However, the scattering process experienced by light before reaching the target area will weaken the correlation between the object light path and the reference light, thereby reducing the imaging quality. In other words, correlative imaging suppresses the subsequent scattering, which is equivalent to making the subsequent scattering medium transparent.
发明内容Contents of the invention
本发明的目的是要解决生物软组织内光学成像深度和分辨率受限于散射的问题，提供一种关联层析装置和方法，以实现生物软组织内，尤其是在散射弥散区域内的光学分辨成像，并且要求该成像方法是无损的。The purpose of the present invention is to solve the problem that the depth and resolution of optical imaging in biological soft tissue are limited by scattering, and provide a correlation tomography device and method to realize optically resolved imaging in biological soft tissue, especially in the scattering diffuse region , and the imaging method is required to be lossless.
本发明的技术解决方案如下：Technical solution of the present invention is as follows:
一种关联层析方法，特点在于该方法包括下列步骤：A correlation chromatography method is characterized in that the method comprises the following steps:
①固定样品；① Fix the sample;
②激光器发出一束激光照射到样品上，同时，超声波发射器发出的超声也作用到样品上，位相共轭镜记录从样品出射的经过光声调制的散射光波前；②The laser emits a beam of laser light to irradiate the sample, and at the same time, the ultrasound emitted by the ultrasonic transmitter also acts on the sample, and the phase conjugate mirror records the scattered light wavefront emitted from the sample after photoacoustic modulation;
③从热光源或赝热光源出射的一束物光经过位相共轭镜反射后照射到样品上，经样品内传播后再出射，出射光用光强探测器探测，获得第一组信号光；③A beam of object light emitted from a thermal light source or a pseudothermal light source is reflected by a phase conjugate mirror and irradiates the sample, and then emerges after propagating in the sample. The outgoing light is detected by a light intensity detector to obtain the first set of signal light;
④从热光源或者赝热源发出的另一束参考光用一个有空间分辨能力的探测器探测获得第二组信号光；或者根据已知的赝热源统计分布计算获得第二组信号光；④ Another beam of reference light emitted from a thermal light source or a pseudo-heat source is detected by a detector with spatial resolution to obtain a second set of signal light; or the second set of signal light is obtained through calculation based on the known statistical distribution of a pseudo-heat source;
⑤两组信号进行关联运算给出关联层析图像。⑤ Correlation operation is performed on the two groups of signals to obtain a correlation tomographic image.
用于上述的关联层析方法的关联层析装置，特点在于其构成包括超声波发射器、位相共轭镜、激光器、光强探测器、空间分辨能力的探测器、热光源或赝热光源和强度关联运算器，所述的激光器发出的激光照射到样品上，所述的超声波发射器发出的超声也作用到样品上，在样品内超声焦点处超声和散射的激光发生声光相互作用，调制光的频率获得频率调制光，从样品出射的频率调制光照射到位相共轭镜上，所述的热光源或者赝热源发出的光被分束器分为透射光和反射光，在所述的透射光方向是所述的空间分辨能力的探测器，所述的反射光束方向是所述的位相共轭镜，在所述的样品外还有光强探测器，所述的光强探测器和空间分辨能力的探测器的输出端与关联运算器的输入端相连。The correlation tomography device used for the above-mentioned correlation tomography method is characterized in that its composition includes an ultrasonic transmitter, a phase conjugate mirror, a laser, a light intensity detector, a detector of spatial resolution, a thermal light source or a pseudothermal light source and an intensity Associative operator, the laser light emitted by the laser is irradiated on the sample, and the ultrasonic wave emitted by the ultrasonic transmitter is also applied to the sample, and the ultrasonic wave and the scattered laser light interact with each other at the focal point of the ultrasonic wave in the sample to modulate the light frequency to obtain frequency modulated light, the frequency modulated light emitted from the sample is irradiated on the phase conjugate mirror, the light emitted by the thermal light source or the pseudo thermal source is divided into transmitted light and reflected light by the beam splitter, and in the transmitted The light direction is the detector with spatial resolution, the reflected beam direction is the phase conjugate mirror, and there is a light intensity detector outside the sample, and the light intensity detector and the space The output of the resolving power detector is connected to the input of the correlation operator.
所述的激光波长范围包含可见光和近红外。Said laser wavelength range includes visible light and near infrared.
所述的热光或者赝热光的波长与频率调制光相同。The wavelength of the thermal light or pseudo-thermal light is the same as that of the frequency modulated light.
所述的超声波的波长范围在1MHz-150MHz之间，超声波发射器的出射端口，即超声转换器的口径和焦长可以选择。The wavelength range of the ultrasonic wave is between 1MHz-150MHz, and the output port of the ultrasonic transmitter, that is, the aperture and focal length of the ultrasonic converter can be selected.
所述的激光、超声、热光源或者赝热光源分别有连续和脉冲两种模式可选，根据需要进行组合，选择脉冲方式时，需要考虑仪器间的同步问题。The laser, ultrasound, thermal light source or pseudothermal light source are respectively available in two modes, continuous mode and pulsed mode, which can be combined according to needs. When selecting the pulsed mode, it is necessary to consider the synchronization problem between instruments.
所述的位相共轭镜的选择范围包含数字式位相共轭镜、光折射晶体、光折射聚合物或其他新型材料共轭镜，The selection range of the phase conjugate mirrors includes digital phase conjugate mirrors, photorefractive crystals, photorefractive polymers or other new material conjugate mirrors,
所述的激光器和位相共轭镜在样品的两对侧或同一侧。The laser and the phase conjugate mirror are on two opposite sides or the same side of the sample.
所述的热光源是统计涨落已知的赝热源，则所述的空间分辨能力的探测器省去。The thermal light source is a pseudo-thermal source with known statistical fluctuations, so the detector with spatial resolution is omitted.
所述的位相共轭镜和反射强度随机调制机构在一起。The phase-conjugate mirror and the reflection intensity random modulation mechanism are combined together.
本发明的技术效果：Technical effect of the present invention:
1、由于非辐射安全并且携带生化信息，生物软组织内的光学成像一直是研究的热点。但目前主要的技术手段都无法同时在成像深度、分辨率和探测效率等方面给出理想的答案。本发明使组织内的成像的分辨率达到光学分辨率，实现了软组织内的光学显微。由于光学信号含有生化信息，关联层析不仅可以直接对软组织内部的结构成像，还含有功能信息，可以用于虚拟光学活检，帮助分辨疾病的生理和病理条件(Chen-Kun Tsai et al.Virtual optical biopsy of human adipocytes with thirdharmonic generation microscopy.Biomedical Optics Express,4,178(2013).)。结合染料、荧光团、报告基因等标记物质，更可以对活体生物组织的功能和代谢过程进行研究。1. Due to its non-radiation safety and carrying biochemical information, optical imaging in biological soft tissues has always been a research hotspot. However, the current main technical means cannot give ideal answers in terms of imaging depth, resolution and detection efficiency at the same time. The invention makes the resolution of the imaging in the tissue reach the optical resolution, and realizes the optical microscope in the soft tissue. Since the optical signal contains biochemical information, correlation tomography can not only directly image the internal structure of soft tissue, but also contain functional information, which can be used in virtual optical biopsy to help distinguish the physiological and pathological conditions of the disease (Chen-Kun Tsai et al.Virtual optical biopsy of human adipocytes with thirdharmonic generation microscopy. Biomedical Optics Express, 4, 178(2013).). Combined with markers such as dyes, fluorophores, and reporter genes, the functions and metabolic processes of living biological tissues can be studied.
2、生物软组织以外，关联层析也可以应用到胶体物质、浑浊的水体等准稳态散射和湍流介质的研究中，提供了强有力的工具。这种方法不使用有辐射的射线、无辐射危害、成本低廉，在临床和实验室有着广泛的应用前景。2. In addition to biological soft tissues, correlation chromatography can also be applied to the study of quasi-steady-state scattering and turbulent media such as colloidal substances and turbid water bodies, providing a powerful tool. This method does not use radiation rays, has no radiation hazards, and has low cost, and has wide application prospects in clinics and laboratories.
附图说明Description of drawings
图1是超声辅助位相共轭光学层析原理图。Figure 1 is a schematic diagram of ultrasound-assisted phase conjugate optical tomography.
图2是数字式位相共轭镜原理，a)波前记录，b)波前反演。Figure 2 is the principle of digital phase conjugate mirror, a) wavefront recording, b) wavefront inversion.
图3是本发明关联层析装置示意图。Fig. 3 is a schematic diagram of the correlation chromatography device of the present invention.
图4是光敏材料共轭镜原理，a)波前记录，b)波前反演。Figure 4 shows the principle of the conjugate mirror of the photosensitive material, a) wavefront recording, b) wavefront inversion.
图5是本发明关联层析方法的流程图。Fig. 5 is a flowchart of the correlation chromatography method of the present invention.
图6是单臂赝热源关联层析原理图。Fig. 6 is a schematic diagram of single-arm pseudothermal source correlation tomography.
图7是半反射式关联层析原理图。Fig. 7 is a schematic diagram of semi-reflective correlation tomography.
图8是全反射式关联层析原理图。Fig. 8 is a schematic diagram of total reflection correlation tomography.
图9是波前共轭和赝热随机耦合的关联层析原理图。Fig. 9 is a schematic diagram of correlation tomography of wavefront conjugation and pseudothermal random coupling.
图10是透射式关联层析实施例结构图。Fig. 10 is a structural diagram of an embodiment of transmission correlation chromatography.
图11是单臂关联层析实施例结构图。Fig. 11 is a structural diagram of an embodiment of single-arm correlation chromatography.
图中：1–样品 2–超声波发射器 201–超声焦点 202–函数发生器 203–信号放大器 204–超声转换器 3–位相共轭镜 4–激光器 401–入射激光束402–入射激光在样品内的弥散光 403–调制散射光 404–位相共轭光或含位相共轭信息的热光/赝热光 405–滤波后的荧光 406–参考激光束 5–光强探测器 6–有空间分辨能力的光强探测器 7–分束器 8–统计涨落光源，如热光源、赝热源等 9–出射光束 10–强度关联运算器 11–光折射晶体 12–共轭参考光束 13–复合共轭和伪随机反射镜 14–双色分束器15－滤波片 16–透镜In the figure: 1-sample 2-ultrasonic emitter 201-ultrasonic focus 202-function generator 203-signal amplifier 204-ultrasonic converter 3-phase conjugate mirror 4-laser 401-incident laser beam 402-incident laser in the sample Diffuse light 403–modulated scattered light 404–phase conjugate light or thermo-optic/pseudo-thermal light with phase conjugate information 405–filtered fluorescence 406–reference laser beam 5–intensity detector 6–spatial resolution capability 7-beam splitter 8-statistical fluctuation light source, such as thermal light source, pseudo-thermal source, etc. 9-exit beam 10-intensity correlation calculator 11-photorefractive crystal 12-conjugate reference beam 13-compound conjugate And pseudo-random mirror 14 - dichroic beam splitter 15 - filter 16 - lens
具体实施方式Detailed ways
以下结合附图和实施例，对本发明作进一步说明，但不应以此限制本发明的保护范围，关联层析的基本原理如图3所示。The present invention will be further described below in conjunction with the accompanying drawings and examples, but the protection scope of the present invention should not be limited by this. The basic principle of correlation chromatography is shown in FIG. 3 .
先请参阅图5，图5是本发明关联层析方法的流程图。由图可见，本发明关联Please refer to FIG. 5 first. FIG. 5 is a flow chart of the correlation chromatography method of the present invention. As can be seen from the figure, the present invention relates
层析方法，该方法包括下列步骤：A chromatographic method comprising the steps of:
①固定样品1；① Fixed sample 1;
②激光器4发出一束激光照射到样品1上，同时，超声波发射器2发出的超声也作用到样品上，位相共轭镜3记录从样品出射的经过光声调制的散射光波前；②The laser 4 emits a beam of laser light to irradiate the sample 1, and at the same time, the ultrasound emitted by the ultrasonic transmitter 2 also acts on the sample, and the phase conjugate mirror 3 records the scattered light wavefront emitted from the sample through photoacoustic modulation;
③从热光源或赝热光源8出射的一束物光经过位相共轭镜3反射后照射到样品1上，经样品1内传播后再出射，出射光用光强探测器5探测，获得第一组信号光；③ A beam of object light emitted from the thermal light source or pseudothermal light source 8 is reflected by the phase conjugate mirror 3 and then irradiates on the sample 1, propagates through the sample 1 and then emerges, and the emitted light is detected by the light intensity detector 5 to obtain the first a set of signal lights;
④从热光源或者赝热源发出的另一束参考光用一个有空间分辨能力的探测器6探测获得第二组信号光；或者根据已知的赝热源统计分布计算获得第二组信号光；④ Another beam of reference light emitted from a thermal light source or a pseudo-heat source is detected by a detector 6 with spatial resolution to obtain a second set of signal light; or the second set of signal light is obtained through calculation based on the known statistical distribution of a pseudo-heat source;
⑤两组信号经关联运算给出关联层析图像。⑤The two groups of signals are correlated tomographic images through correlative operation.
参阅图3，图3是本发明关联层析装置示意图。由图可见，本发明关联层析装置，构成包括超声波发射器2、位相共轭镜3、激光器4、光强探测器5、空间分辨能力的探测器6、热光源或赝热光源8和强度关联运算器10，所述的激光器4发出的激光照射到样品1上，所述的超声波发射器2发出的超声也作用到样品1上，在样品1内超声焦点201处超声和散射的激光发生声光相互作用，调制光的频率获得频率调制光，从样品出射的频率调制光照射到位相共轭镜3上，所述的热光源或者赝热源8发出的光被分束器7分为透射光和反射光，在所述的透射光方向是所述的空间分辨能力的探测器6，所述的反射光束方向是所述的位相共轭镜3，在所述的样品1外还有光强探测器5，所述的光强探测器5和空间分辨能力的探测器6的输出端与关联运算器10的输入端相连。Referring to Fig. 3, Fig. 3 is a schematic diagram of the correlation chromatography device of the present invention. It can be seen from the figure that the correlative tomography device of the present invention comprises an ultrasonic transmitter 2, a phase conjugate mirror 3, a laser 4, a light intensity detector 5, a detector 6 of spatial resolution, a thermal light source or a pseudothermal light source 8 and an intensity The associated computing unit 10, the laser light emitted by the laser 4 is irradiated on the sample 1, the ultrasonic wave emitted by the ultrasonic transmitter 2 is also applied to the sample 1, and the ultrasonic and scattered laser light are generated at the ultrasonic focal point 201 in the sample 1 Acousto-optic interaction, modulate the frequency of the light to obtain frequency-modulated light, the frequency-modulated light emitted from the sample is irradiated on the phase conjugate mirror 3, and the light emitted by the thermal light source or pseudo-thermal source 8 is split by the beam splitter 7 for transmission Light and reflected light, the direction of the transmitted light is the detector 6 of the spatial resolution, the direction of the reflected light beam is the phase conjugate mirror 3, and there is light outside the sample 1 The intensity detector 5 , the output terminals of the light intensity detector 5 and the spatial resolution detector 6 are connected to the input terminal of the correlation operator 10 .
所述的激光波长范围包含可见光和近红外。Said laser wavelength range includes visible light and near infrared.
所述的热光或者赝热光的波长与频率调制光相同。The wavelength of the thermal light or pseudo-thermal light is the same as that of the frequency modulated light.
所述的超声波的波长范围在1MHz-150MHz之间，超声波发射器的出射端口，即超声转换器的口径和焦长可以选择。The wavelength range of the ultrasonic wave is between 1MHz-150MHz, and the output port of the ultrasonic transmitter, that is, the aperture and focal length of the ultrasonic converter can be selected.
所述的激光、超声、热光源或者赝热光源分别有连续和脉冲两种模式可选，根据需要进行组合，选择脉冲方式时，需要考虑仪器间的同步问题。The laser, ultrasound, thermal light source or pseudothermal light source are respectively available in two modes, continuous mode and pulsed mode, which can be combined according to needs. When selecting the pulsed mode, it is necessary to consider the synchronization problem between instruments.
所述的位相共轭镜的选择范围包含数字式位相共轭镜、光折射晶体、光折射聚合物或其他新型材料共轭镜，The selection range of the phase conjugate mirrors includes digital phase conjugate mirrors, photorefractive crystals, photorefractive polymers or other new material conjugate mirrors,
在图3中，从激光器4发出的激光照射到样品1上，同时从超声波发射器2发出的超声也作用到样品1上。在样品1内超声焦点201处超声和散射的激光发生声光相互作用，从而调制光的频率获得频率调制光。从样品出射的频率调制光照射到位相共轭镜3上，从而波前被记录。随后，从热光源或者赝热源8发出的光被分束器7分为透射光和反射光，所述的透射光(称为参考束)由有空间分辨能力的探测器6(如CCD相机)探测得到参考光强信号，所述的反射光束照射到位相共轭镜3，经过位相共轭镜3反射后照射到样品1上。在样品1外用光强探测器5探测出射光的光强信号，该光强信号与参考光强信号进行关联运算给出层析图像。In FIG. 3 , the laser light emitted from the laser 4 is irradiated on the sample 1 , and the ultrasonic wave emitted from the ultrasonic transmitter 2 is also applied to the sample 1 . Acousto-optic interaction occurs between the ultrasonic wave and the scattered laser light at the ultrasonic focal point 201 in the sample 1, thereby modulating the frequency of the light to obtain frequency-modulated light. The frequency-modulated light emitted from the sample is irradiated onto the phase conjugate mirror 3, so that the wavefront is recorded. Subsequently, the light emitted from the thermal light source or pseudo-thermal source 8 is divided into transmitted light and reflected light by a beam splitter 7, and the transmitted light (referred to as a reference beam) is detected by a spatially resolving detector 6 (such as a CCD camera) The reference light intensity signal is obtained by detection, and the reflected light beam irradiates the phase conjugate mirror 3 , and then irradiates the sample 1 after being reflected by the phase conjugate mirror 3 . A light intensity detector 5 is used outside the sample 1 to detect the light intensity signal of the outgoing light, and the light intensity signal is correlated with the reference light intensity signal to obtain a tomographic image.
其中，入射激光波长范围包含可见光和近红外，热光或者赝热光的波长与频率调制光相同。超声波的波长范围在1MHz-150MHz之间，超声波发射器的出射端口，即超声转换器的口径和焦长可以选择。激光、超声和热光源或者赝热光源分别有连续和脉冲两种模式可选，根据需要进行组合。选择脉冲方式时，需要考虑仪器间的同步问题。位相共轭镜3的选择范围包含数字式位相共轭镜、光折射晶体、光折射聚合物或其他新型材料共轭镜，如图4所示。Wherein, the incident laser wavelength range includes visible light and near infrared, and the wavelength of thermal light or pseudothermal light is the same as that of frequency modulated light. The wavelength range of the ultrasonic wave is between 1MHz-150MHz, and the exit port of the ultrasonic transmitter, that is, the aperture and focal length of the ultrasonic converter can be selected. Laser, ultrasonic and thermal light sources or pseudothermal light sources are available in continuous and pulsed modes, which can be combined according to needs. When selecting the pulse mode, the synchronization between instruments needs to be considered. The selection range of the phase conjugate mirror 3 includes digital phase conjugate mirrors, photorefractive crystals, photorefractive polymers or other new material conjugate mirrors, as shown in FIG. 4 .
超声焦点的定位可以借助超声扫描样品所得到的低分辨三维图像。对比较大的感兴趣区层析，可以通过改变样品和超声焦点的相对位置，多次重复焦点层析来实现。显然，扫描超声焦点，可以实现整个样品的三维层析。The location of the ultrasonic focus can be obtained by means of a low-resolution three-dimensional image obtained by ultrasonically scanning the sample. Comparing the tomography of a larger region of interest can be achieved by changing the relative position of the sample and the ultrasound focus and repeating the focus tomography multiple times. Apparently, by scanning the ultrasound focus, three-dimensional tomography of the entire sample can be achieved.
采用赝热源无参考光束结构时，图3可以简化为图6。其中，光源8是统计涨落已知的赝热源。图3和图6中，激光入射点和位相共轭镜位于样品两侧，这是透射式结构，也可采用图7所示的半反射式结构，即激光入射点和位相共轭镜在样品同一侧；或者图8所示全反射式结构，即所有设备都在样品的同一侧。位相共轭镜和赝热源可以进一步整合到一起，例如在位相共轭的SLM上叠加上反射强度随机调制机构。关联层析的装置可以进而简化为图9。Figure 3 can be simplified to Figure 6 when a pseudo-heat source without a reference beam structure is used. Among them, the light source 8 is a pseudo heat source whose statistical fluctuation is known. In Figure 3 and Figure 6, the laser incident point and phase conjugate mirror are located on both sides of the sample, which is a transmissive structure, and the semi-reflective structure shown in Figure 7 can also be used, that is, the laser incident point and phase conjugate mirror are on the sample. The same side; or a total reflection structure as shown in Figure 8, that is, all devices are on the same side of the sample. The phase-conjugate mirror and the pseudo-heat source can be further integrated, for example, superimposing a reflection intensity random modulation mechanism on the phase-conjugate SLM. The device for correlation chromatography can be further simplified as shown in Fig. 9 .
实施例1：Example 1:
如图10，其中，超声波发射器2由超声转换器204、信号发生器202和信号放大器203构成。脉冲激光器4(例如，Coherent passively Q-switched laser FLARE,波长范围390nm-800nm)发出的脉冲激光401和超声转换器204发出的脉冲超声(例如，1MHz的一个或多个周期，10MHz的一个或多个周期，150MHz的一个或多个周期)同时作用到样品1上，PC或者时延发生器(图中未标出)控制两个脉冲信号的时延使激光和超声同时到达超声焦点201(或者更准确到焦点内的不同位置)。超声转换器由函数发生器202和信号放大器203驱动。样品外，出射的频率调制光403照射到位相共轭镜3上被记录。随后，热光源8发出的热光束9照射位相共轭镜3，反射光404进入样品传播后再出射。探测器5探测从样品出射的光强，CCD记录参考臂的光强分布，两组关联信号由强度关联运算器10做关联运算，从而得到层析图像。移动超声转换器扫描焦点，可以给出整个软组织样品的三维光学层析。As shown in FIG. 10 , the ultrasonic transmitter 2 is composed of an ultrasonic converter 204 , a signal generator 202 and a signal amplifier 203 . The pulsed laser 401 sent by the pulsed laser 4 (for example, Coherent passively Q-switched laser FLARE, wavelength range 390nm-800nm) and the pulsed ultrasound sent by the ultrasonic converter 204 (for example, one or more cycles of 1MHz, one or more cycles of 10MHz) cycles, one or more cycles of 150MHz) act on the sample 1 at the same time, and the PC or the delay generator (not shown in the figure) controls the time delay of the two pulse signals so that the laser and the ultrasound arrive at the ultrasound focus 201 (or more accurately to different positions within the focal point). The ultrasonic transducer is driven by a function generator 202 and a signal amplifier 203 . Outside the sample, the outgoing frequency modulated light 403 is irradiated onto the phase conjugate mirror 3 to be recorded. Subsequently, the thermal light beam 9 emitted by the thermal light source 8 irradiates the phase conjugate mirror 3 , and the reflected light 404 enters the sample, propagates and then exits. The detector 5 detects the light intensity emitted from the sample, the CCD records the light intensity distribution of the reference arm, and the two sets of correlation signals are correlated by the intensity correlation calculator 10 to obtain a tomographic image. By moving the scanning focus of the ultrasound transducer, a three-dimensional optical tomography of the entire soft tissue sample can be given.
实施例2：如图11所示，采用相对简化的光路和探测，光源8用计算机控制的SLM或者数字微镜器件DMD(digital micromirror device,简称为DMD)反射激光(可以从入射光光束401分束而来，或者另一台激光器发射)取代，根据伪随机数列调节SLM或者DMD上的像素的分布从而产生赝热光。出射的赝热光经过位相共轭镜3反射，反射光照射样品1。探测器5探测从样品1出射的光，由于赝热源的伪随机涨落已知，可以计算得到参考臂的光强分布，两组数据在计算机做关联后得到层析图像。Embodiment 2: as shown in Figure 11, adopt relatively simplified optical path and detection, SLM or digital micromirror device DMD (digital micromirror device, be called for short DMD) reflection laser of light source 8 with computer control (can be divided from incident light beam 401 Beam from, or another laser emission) instead, adjust the distribution of pixels on the SLM or DMD according to the pseudo-random sequence to generate pseudo-thermal light. The outgoing pseudothermal light is reflected by the phase conjugate mirror 3, and the reflected light illuminates the sample 1. The detector 5 detects the light emitted from the sample 1. Since the pseudo-random fluctuation of the pseudo-heat source is known, the light intensity distribution of the reference arm can be calculated, and the two sets of data are correlated by a computer to obtain a tomographic image.
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Application publication date: 20141224 |