WO2014201590A1 - 一种全息荧光分子成像系统及方法 - Google Patents

一种全息荧光分子成像系统及方法 Download PDF

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WO2014201590A1
WO2014201590A1 PCT/CN2013/000726 CN2013000726W WO2014201590A1 WO 2014201590 A1 WO2014201590 A1 WO 2014201590A1 CN 2013000726 W CN2013000726 W CN 2013000726W WO 2014201590 A1 WO2014201590 A1 WO 2014201590A1
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holographic
fluorescence
molecular imaging
excitation light
imaging system
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白净
左思敏
刘飞
何耘
曹良才
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Tsinghua University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N2021/6417—Spectrofluorimetric devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N2021/6417—Spectrofluorimetric devices
    • G01N2021/6423—Spectral mapping, video display

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  • the present invention relates to a fluorescent molecular imaging system and method, and more particularly to a holographic fluorescent molecular imaging system and method using a holographic grating as a spatial filtering and/or spectral filter. Background technique
  • Fluorescent molecular imaging technology is an emerging molecular imaging technology that has developed rapidly in recent years. It has broad application prospects in the fields of tumor detection, drug development and disease diagnosis. Fluorescent molecular imaging technology uses fluorescent markers to label specific molecules or cells in small animals. When the labeled small animals are irradiated with excitation light of appropriate wavelength and intensity, the fluorescent markers in the small animals are excited to emit fluorescence. A certain device detects the intensity of the fluorescence produced, and a distribution image of the fluorescence optical properties inside the tissue can be obtained, thereby allowing spatial and temporal visual description of normal or abnormal biological processes at the molecular and cellular levels. Fluorescence Molecular imaging is a highly sensitive, ion-free, non-invasive, and low-cost imaging modality.
  • the fluorescence filtering of the existing fluorescent molecular imaging system mostly uses spectral dispersion filters such as ordinary optical filters and liquid crystal tunable filters (LCTF), and only one-dimensional signal acquisition can obtain two-dimensional, single-spectral fluorescent markers. Information.
  • spectral dispersion filters such as ordinary optical filters and liquid crystal tunable filters (LCTF)
  • LCTF liquid crystal tunable filters
  • an object of the present invention is to provide a holographic fluorescent molecular imaging system and method for realizing rapid two-dimensional or three-dimensional spatial reconstruction and spectral characteristic reconstruction.
  • a holographic fluorescent molecular imaging system comprising an experimental platform for placing an object to be imaged, an excitation light illumination unit disposed on one side of the experimental platform, a CCD detecting device disposed on the other side of the experimental platform, and a computer connected to the excitation light illumination unit and the CCD detecting device, wherein a holographic fluorescent filter is disposed between the experimental platform and the CCD detecting device unit.
  • the holographic fluorescent filtering unit employs a holographic grating as the core optical device.
  • a focusing mirror assembly and an imaging lens are respectively spaced apart from each other on the front and rear sides of the holographic grating according to specific experimental conditions.
  • the excitation light illumination unit includes an excitation light source and a filter wheel.
  • the experimental platform is a 360° rotating test bench.
  • a holographic fluorescent molecular imaging method using the above system comprising the following steps:
  • Step 1 placing the object to be imaged on the experimental platform
  • Step 2 the excitation light emitted by the excitation light illumination unit propagates and illuminates the object to be imaged, and the generated fluorescent signal passes through the holographic fluorescence filtering unit to realize spatial filtering and/or spectral filtering;
  • Step 3 The CCD detection device directly obtains two-dimensional or three-dimensional fluorescent marker distribution images and rich multi-spectral fluorescence information, and sends them to a computer for data processing.
  • the holographic fluorescent filtering unit employs a holographic grating as the core optical device.
  • step 2 the desired holographic grating is determined and selected according to specific experimental conditions, and a focusing mirror assembly and an imaging lens are respectively disposed at intervals on the front and rear sides of the holographic grating.
  • the imaging method employs transmissive fluorescence imaging or reflective fluorescence imaging.
  • the present invention has the following advantages due to the above technical solution:
  • the present invention provides a holographic fluorescence filtering unit between the small animal experimental platform and the CCD detecting device as a spatial filtering and spectral filter component, so that the CCD detecting device can directly obtain three-dimensional at the same time.
  • Fluorescent markers spatially distribute images and rich multi-spectral fluorescence information, eliminating the need for complex image reconstruction algorithms and time-consuming complex scans.
  • Figure 1 is a schematic view of the structural block of the present invention. detailed description
  • a holographic fluorescent molecular imaging system of the present invention which comprises a small animal experimental platform 1, an excitation light illumination unit 2 disposed on one side of a small animal experiment platform 1, and a CCD disposed on the other side of the small animal experiment platform 1.
  • the present invention differs from existing fluorescent molecular imaging systems in that a holographic fluorescent filtering unit 4 is used in place of a conventional fluorescent filter as a fluorescent space and spectral filter element of a fluorescent molecular imaging system.
  • the holographic fluorescent filter unit 4 is a set of optical components having the hologram grating 41 as a core.
  • a focusing mirror assembly 42 and an imaging lens 43 are respectively disposed on the front and rear sides of the hologram grating 41, wherein the focusing mirror assembly 42 includes a plurality of focusing mirrors.
  • the excitation light illumination unit 2 includes an excitation light source 21 and a filter wheel 22, wherein the filter wheel 22 is used to control the excitation light wavelength of the excitation light source 21.
  • the small animal experimental platform 1 can be a 360° rotating test bench.
  • the present invention also provides a holographic fluorescent molecular imaging method comprising the following steps:
  • Step 1 Place the object to be imaged 6 on the small animal experiment platform 1.
  • the desired holographic grating 41 is determined and selected according to specific experimental conditions such as experimental purpose, experimental animal and fluorescent label. For example, when performing multi-spectral fluorescence molecular imaging experiments, it is necessary to simultaneously image a plurality of wavelengths (1 ⁇ 2, ⁇ 2 ⁇ ... fluorescent signals, in which case a holographic multi-spectral dispersion device having ⁇ . spectral region segments can be selected as the holographic grating 41, Multi-channel separation and fast imaging for multi-spectral fluorescence images.
  • Step 3 the excitation light emitted by the excitation light source 21 passes through the filter wheel 22, propagates and illuminates the object to be imaged 6, and the generated fluorescent signal is spatially filtered and/or spectrally filtered by the holographic fluorescence filtering unit 4 to realize spatial decomposition of the fluorescent signal. And / or spectral decomposition.
  • Step 4 The CCD detecting device 3 directly obtains a spatially distributed image of the two-dimensional or three-dimensional fluorescent marker and rich multi-spectral fluorescence information, and sends the data to the computer 4 for data processing, thereby eliminating the need for complicated image reconstruction algorithms and consumption. Complex scans of time.
  • the fluorescence imaging mode can be either transmissive fluorescence imaging or retroreflective fluorescence imaging.
  • transmissive fluorescent molecular imaging refers to the excitation and detection of fluorescence on the same surface
  • transmissive fluorescent molecular imaging is to stimulate and detect fluorescence on different surfaces.

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  • Health & Medical Sciences (AREA)
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Abstract

一种全息荧光分子成像系统及方法，该系统包括一用来放置待成像物体的实验平台（1），一设置于所述实验平台一侧的激发光照明单元（2），一设置于所述实验平台另一侧的CCD检测装置（3），以及一连接所述激发光照明单元（2）和CCD检测装置（3）的计算机（5），在所述实验平台（1）和CCD检测装置（3）之间设置有一个全息荧光滤波单元（4）。该系统采用全息荧光滤波单元（4）作为空间滤波和光谱滤波器件，从而使得CCD检测装置（3）可以同时直接获得二维或三维的荧光标记物的空间分布影像和丰富的多光谱荧光信息，因而不需要复杂的图像重建算法和耗时的复杂扫描。

Description

一种全息荧光分子成像系统及方法 技术领域
本发明涉及一种荧光分子成像系统及方法， 特别是关于一种利用全息光栅来作 为空间滤波和 /或光谱滤波器的全息荧光分子成像系统及方法。 背景技术
荧光分子成像技术是近年发展迅速的一种新兴的分子成像技术， 在肿瘤检测、 药物研发和疾病诊断等领域有着广阔的应用前景。 荧光分子成像技术是利用荧光标 记物标记小动物体内的特定分子或细胞， 采用合适波段和强度的激发光照射被标记 的小动物时， 小动物体内的荧光标记物受到激发从而发出荧光， 通过采用一定的装 置检测产生的荧光强度， 就可以获得组织内部荧光光学特性的分布图像， 从而可以 从分子和细胞水平上对正常或异常的生物过程进行空间和时间上的视觉描述。 荧光 分子成像技术是一种高灵敏度、 无电离辐射、 非侵入式和低成本的成像方式。
但是， 现有的荧光分子成像系统的荧光滤波多采用普通光学滤光片、 液晶可调 谐滤波器 （LCTF ) 等光谱色散滤波器件， 单次信号采集只能获得二维、 单光谱的荧 光标记物的信息。 这使得当系统需要重建出立体的、 多光谱的荧光图像时， 就必须 进行多次扫描且采用复杂的图像重建算法， 因此导致现有的荧光分子成像系统耗时 长、 时间分辨率差、 光谱波段少。 而在实际应用中， 常常需要实现快速的荧光信号 的空间、 光谱、 时间信号的采集， 尤其是针对一些快速的、 动态变化的药物代谢和 多荧光标记物的成像分析， 现有的荧光分子成像系统不能满足要求。 发明内容
针对上述问题， 本发明的目的是提供一种全息荧光分子成像系统及方法， 以实 现快速的二维或三维空间重建和光谱特性重建。
为实现上述目的， 本发明采取以下技术方案： 一种全息荧光分子成像系统， 该 系统包括一用来放置待成像物体的实验平台， 一设置于所述实验平台一侧的激发光 照明单元， 一设置于所述实验平台另一侧的 CCD检测装置， 以及一连接所述激发光 照明单元和 CCD检测装置的计算机， 其特征在于， 在所述实验平台和 CCD检测装置 之间设置有一全息荧光滤波单元。 在一个优选的实施例中， 所述全息荧光滤波单元 采用全息光栅作为核心光学器件。 在一个优选的实施例中， 根据具体实验条件， 在 所述全息光栅的前后两侧分别间隔设置一聚焦镜组件和一成像透镜。 在一个优选的实施例中， 所述激发光照明单元包括激发光源和滤光片轮。 在一个优选的实施例中， 所述实验平台为 360° 旋转实验台。
一种采用上述系统的全息荧光分子成像方法， 其包括以下步骤：
步骤 1， 将待成像物体放置在实验平台上；
步骤 2， 激发光照明单元发射的激发光传播并照射待成像物体， 产生的荧光信 号经过全息荧光滤波单元， 实现空间滤波和 /或光谱滤波；
步骤 3， CCD检测装置同时直接获得二维或者三维的荧光标记物分布影像和丰 富的多光谱荧光信息， 并将其发送到计算机进行数据处理。
在一个优选的实施例中， 所述全息荧光滤波单元采用全息光栅作为核心光学器 件。
在进行步骤 2时， 根据具体实验条件， 确定并选择所需要的全息光栅， 以及在 所述全息光栅的前后两侧分别间隔设置一聚焦镜组件和一成像透镜。
在一个优选的实施例中， 该成像方法采用透射式荧光成像或反射式荧光成像。 本发明由于采取以上技术方案， 其具有以下优点： 本发明在小动物实验平台和 CCD检测装置之间设置一全息荧光滤波单元作为空间滤波和光谱滤波器件， 从而使 得 CCD检测装置可以同时直接获得三维的荧光标记物空间分布影像和丰富的多光谱 荧光信息， 因而不需要复杂的图像重建算法和耗时的复杂扫描。 附图说明
以下结合附图来对本发明进行详细的描绘。 然而应当理解， 附图的提供仅为了 更好地理解本发明， 它们不应该理解成对本发明的限制。
图 1是本发明的结构框示意图。 具体实施方式
下面结合附图和实施例对本发明进行详细的描述。
图 1显示了本发明的全息荧光分子成像系统， 该系统包括小动物实验平台 1， 设置于小动物实验平台 1一侧的激发光照明单元 2， 设置于小动物实验平台 1另一 侧的 CCD检测装置 3， 设置于 CCD检测装置 3和小动物实验平台 1之间的全息荧光 滤波单元 4， 以及分别与激发光照明单元 2和 CCD检测装置 3连接的计算机 5。
本发明与现有的荧光分子成像系统的区别在于用全息荧光滤波单元 4代替普通 荧光滤光片来作为荧光分子成像系统的荧光空间和光谱滤波器件。 在本实施例中， 全息荧光滤波单元 4是一组以全息光栅 41为核心的光学组件。 同时还可以根据具 体实验条件， 在全息光栅 41的前后两侧分别设置有一聚焦镜组件 42和一成像透镜 43， 其中聚焦镜组件 42包含若干聚焦镜。
在一个优选的实施例中， 激发光照明单元 2包括激发光源 21和滤光片轮 22， 其中滤光片轮 22用以控制激发光源 21的激发光波长。
在一个优选实施例中， 小动物实验平台 1可以是一个 360° 旋转实验台。
根据上述各实施例中提供的全息荧光分子成像系统， 本发明还提出了一种全息 荧光分子成像方法， 其包括以下步骤：
步骤 1， 将待成像物体 6放置在小动物实验平台 1上。
步骤 2， 根据实验目的、 实验动物和荧光标记物等具体实验条件， 确定并选择 所需要的全息光栅 41。例如进行多光谱荧光分子成像实验时， 需要同时对多个波长 ( ½, λ2ί… 的荧光信号进行成像， 此时可以选用具有 η.个光谱区分段的全息多 光谱色散器件作为全息光栅 41，对多光谱荧光影像实现一次多通道分离和快速成 像。
步骤 3，激发光源 21发射的激发光通过滤光片轮 22，传播并照射待成像物体 6， 产生的荧光信号经过全息荧光滤波单元 4 的空间滤波和 /或光谱滤波， 实现荧光信 号的空间分解和 /或光谱分解。
步骤 4， CCD检测装置 3 同时直接获得二维或三维的荧光标记物空间分布影像 和丰富的多光谱荧光信息， 并将其发送到计算机 4进行数据处理， 因而不需要复杂 的图像重建算法和耗时的复杂扫描。
在一个优选的实施例中， 荧光成像方式既可以是透射式荧光成像， 也可以是反 射式荧光成像。 其中， 反射式荧光分子成像是指在同一表面上对荧光进行激发和检 测， 透射式荧光分子成像则是在不同表面上对荧光进行激发和检测。
上述各实施例仅用于对本发明的目的、 技术方案和有益效果进行了进一步详细 说明， 并不用于限制本发明， 凡在本发明的精神和原则之内， 所做的任何修改、 等 同替换、 改进等， 均应包含在本发明的保护范围之内。

Claims

权利要求
1、 一种全息荧光分子成像系统， 该系统包括一用来放置待成像物体的实验平 台， 一设置于所述实验平台一侧的激发光照明单元， 一设置于所述实验平台另一侧 的 CCD检测装置， 以及一连接所述激发光照明单元和 CCD检测装置的计算机， 其特 征在于， 在所述实验平台和 CCD检测装置之间设置有一全息荧光滤波单元。
2、 如权利要求 1 所述的一种全息荧光分子成像系统， 其特征在于， 所述全息 荧光滤波单元采用全息光栅作为核心光学器件。
3、 如权利要求 2 所述的一种全息荧光分子成像系统， 其特征在于， 根据具体 实验条件， 在所述全息光栅的前后两侧分别间隔设置一聚焦镜组件和一成像透镜。
4、 如权利要求 1到 3任一项所述的一种全息荧光分子成像系统， 其特征在于， 所述激发光照明单元包括激发光源和滤光片轮。
5、 如权利要求 1到 4任一项所述的一种全息荧光分子成像系统， 其特征在于， 所述实验平台为 360° 旋转实验台。
6、 一种采用如权利要求 1到 5任一项所述系统的全息荧光分子成像方法， 其 包括以下步骤：
步骤 1， 将待成像物体放置在实验平台上；
步骤 2， 激发光照明单元发射的激发光传播并照射待成像物体， 产生的荧光信 号经过全息荧光滤波单元， 实现空间滤波和 /或光谱滤波；
步骤 3， CCD检测装置同时直接获得二维或者三维的荧光标记物分布影像和丰 富的多光谱荧光信息， 并将其发送到计算机进行数据处理。
7、 如权利要求 6所述的全息荧光分子成像方法， 其特征在于， 所述全息荧光 滤波单元采用全息光栅作为核心光学器件。
8、 如权利要求 7 所述的全息荧光分子成像方法， 其特征在于， 在进行步骤 2 时， 根据具体实验条件， 确定并选择所需要的全息光栅， 以及在所述全息光栅的前 后两侧分别间隔设置一聚焦镜组件和一成像透镜。
9、 如权利要求 6到 8任一项所述的全息荧光分子成像方法， 其特征在于， 该 成像方法采用透射式荧光成像或反射式荧光成像。
PCT/CN2013/000726 2013-06-21 2013-06-21 一种全息荧光分子成像系统及方法 Ceased WO2014201590A1 (zh)

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