CN108020509A - The method and its device of a kind of optical projection tomography - Google Patents
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Abstract
Description
技术领域technical field
本发明创造涉及生物成像技术领域，特别涉及一种光学投影层析成像的方法及其装置。The invention relates to the technical field of biological imaging, in particular to an optical projection tomography method and a device thereof.
背景技术Background technique
光学投影层析成像(Optical Projection Tomography，OPT)技术，是一种新的三维光学成像技术，能够实现探测深度达到10mm的小动物的活体三维成像。Optical projection tomography (Optical Projection Tomography, OPT) technology is a new three-dimensional optical imaging technology, which can realize three-dimensional imaging of small animals with a detection depth of 10mm.
而光学投影层析成像它是基于显微镜相机成像，在显微成像过程中，由于显微系统的景深很小，对具有一定厚度的物体成像，得到的图像中只有部分细节良好聚焦，其他部分则是模糊的，这使得人们无法用光学投影层析成像的方法清楚地研究样品的内部结构。对成像系统而言,大的景深意味着同一画面中有更多的清晰景物,意味着更多的可测控、监控对象。因此，增大光学系统的景深问题一直以来都是应用于光学领域的亟待解决的问题。Optical projection tomography is based on microscope camera imaging. In the process of microscopic imaging, due to the small depth of field of the microscopic system, when imaging an object with a certain thickness, only part of the details in the obtained image are in good focus, and other parts are in focus. is blurred, which makes it impossible to clearly study the internal structure of the sample with optical projection tomography. For the imaging system, a large depth of field means that there are more clear scenes in the same picture, which means more objects that can be measured, controlled and monitored. Therefore, increasing the depth of field of an optical system has always been an urgent problem to be solved in the field of optics.
目前，对增大光学系统中景深的方法主要有比如幅度切砋法、二维成像序列的图像融合、特殊设计透镜结合图像处理、带有相位模板的普通镜头结合图像处理(波前编码)、三维数字全息等。幅度切砋法会极大降低成像光能量和成像分辨率；图像融合法和数字全息方法都需要进行耗时较长的复杂运算。At present, the methods for increasing the depth of field in the optical system mainly include the amplitude cutting method, image fusion of two-dimensional imaging sequence, special design lens combined with image processing, common lens with phase template combined with image processing (wavefront encoding), 3D digital holography, etc. The amplitude cutting method will greatly reduce the imaging light energy and imaging resolution; both the image fusion method and the digital holography method require complex calculations that take a long time.
发明内容Contents of the invention
本发明的目的是：提供一种扩大光学投影层析成像景深的方法和装置。The object of the present invention is to provide a method and device for enlarging the field depth of optical projection tomography.
本发明解决其技术问题的解决方案是，一方面：一种光学投影层析成像的方法，包括步骤：拍摄生物样品若干张不同角度不同焦面的图像；利用拉普拉斯金字塔算法将同一个角度不同焦面的图像融合成长景深图片；利用滤波反投影算法将生物样品不同角度的长景深图片重建。The solution of the present invention to solve its technical problems is, on the one hand: a method for optical projection tomography, comprising the steps of: taking several images of biological samples with different angles and different focal planes; The images of different focal planes from different angles are fused into a long depth-of-field picture; the long-field depth-of-field pictures from different angles of biological samples are reconstructed by using the filter back projection algorithm.
进一步，所述步骤获得生物样品若干张不同角度不同焦面的图像中所述不同焦面为5个不同焦面。Further, the step obtains several images of the biological sample with different angles and different focal planes, in which the different focal planes are 5 different focal planes.
进一步，在所述拍摄生物样品若干张不同角度不同焦面的图像前，将所述生物样品放置在充满水的玻璃池中。Further, before taking several images of the biological sample at different angles and with different focal planes, the biological sample is placed in a glass pool filled with water.
另一方面，一种光学投影层析成像的装置，包括：平行光源器、相机、远心成像系统、步进电机、旋转样品台、计算机，所述计算机与所述相机电连，所述平行光源器、旋转样品台、远心成像系统、相机依次通过光路连接，所述步进电机用于带动所述相机朝生物样品直线移动，所述旋转样品台用于带动所述生物样品转动，所述相机用于在步进电机带动下、旋转样品台的转动下，拍摄生物样品若干张不同角度不同焦面的图像，所述计算机用于获取相机拍摄的图像，并利用拉普拉斯金字塔算法将同一个角度不同焦面的图像融合成长景深图片，利用滤波反投影算法将生物样品不同角度的长景深图片组成三维图像。On the other hand, an optical projection tomography device includes: a parallel light source, a camera, a telecentric imaging system, a stepper motor, a rotating sample stage, and a computer, the computer is electrically connected to the camera, and the parallel The light source device, the rotating sample stage, the telecentric imaging system, and the camera are sequentially connected through the optical path. The stepping motor is used to drive the camera to move linearly towards the biological sample, and the rotating sample stage is used to drive the biological sample to rotate. The camera is used to take several images of biological samples with different angles and different focal planes driven by the stepping motor and the rotation of the rotating sample stage, and the computer is used to obtain the images taken by the camera, and use the Laplacian pyramid algorithm The images of the same angle and different focal planes are fused into a long depth of field image, and the long depth of field images of different angles of biological samples are combined into a three-dimensional image by using the filter back projection algorithm.
进一步，所述旋转样品台上设有充满水的玻璃池。Further, a glass pool filled with water is provided on the rotating sample stage.
进一步，所述远心成像系统包括：前、后透镜、光阑，所述光阑位于所述前、后透镜之间，并位于所述前透镜的后焦面上。Further, the telecentric imaging system includes: front and rear lenses, and an aperture, and the aperture is located between the front and rear lenses and on the rear focal plane of the front lens.
进一步，所述相机为COMS相机。Further, the camera is a COMS camera.
本发明的有益效果是：一方面，该方法通过对每一个焦面时通过采集不同焦点的照片，再通过拉普拉斯金字塔算法，从而得出长景深照片，再利用滤波反投影算法进行重建，使得图像所提取的信息充分，图像分辨率高。The beneficial effects of the present invention are: on the one hand, the method collects photos of different focal points for each focal plane, and then uses the Laplacian pyramid algorithm to obtain a long depth of field photo, and then uses the filter back projection algorithm to reconstruct , so that the information extracted from the image is sufficient and the image resolution is high.
另一方面，提供一种基于本发明方法的装置，该装置利用相机、远心成像系统、步进电机、旋转平台对每一个焦面时通过采集不同焦点的照片，再通过计算机的拉普拉斯金字塔算法，从而得出长景深照片，再利用滤波反投影算法进行重建，得到信息充分、分辨率高的图像。On the other hand, provide a kind of device based on the method of the present invention, when this device utilizes camera, telecentric imaging system, stepping motor, rotating platform, by gathering the photograph of different focal point when each focal plane, then by Lapla of computer The pyramid algorithm is used to obtain a long depth of field photo, and then the filtered back projection algorithm is used to reconstruct it to obtain an image with sufficient information and high resolution.
附图说明Description of drawings
为了更清楚地说明本发明实施例中的技术方案，下面将对实施例描述中所需要使用的附图作简单说明。显然，所描述的附图只是本发明的一部分实施例，而不是全部实施例，本领域的技术人员在不付出创造性劳动的前提下，还可以根据这些附图获得其他设计方案和附图。In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly describe the drawings that need to be used in the description of the embodiments. Apparently, the described drawings are only some embodiments of the present invention, not all embodiments, and those skilled in the art can obtain other designs and drawings based on these drawings without creative work.
图1是本发明创造装置的结构示意图；Fig. 1 is the structural representation of creation device of the present invention;
图2是实施例1的光学投影层成像方法的流程图。FIG. 2 is a flow chart of the optical projection layer imaging method in Embodiment 1. FIG.
具体实施方式Detailed ways
以下将结合实施例和附图对本发明的构思、具体结构及产生的技术效果进行清楚、完整地描述，以充分地理解本发明的目的、特征和效果。显然，所描述的实施例只是本发明的一部分实施例，而不是全部实施例，基于本发明的实施例，本领域的技术人员在不付出创造性劳动的前提下所获得的其他实施例，均属于本发明保护的范围。另外，文中所提到的所有联接/连接关系，并非单指构件直接相接，而是指可根据具体实施情况，通过添加或减少联接辅件，来组成更优的联接结构。本发明创造中的各个技术特征，在不互相矛盾冲突的前提下可以交互组合。The idea, specific structure and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments and accompanying drawings, so as to fully understand the purpose, features and effects of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, rather than all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts belong to The protection scope of the present invention. In addition, all the connection/connection relationships mentioned in this article do not refer to the direct connection of components, but mean that a better connection structure can be formed by adding or reducing connection accessories according to specific implementation conditions. The various technical features in the invention can be combined interactively on the premise of not conflicting with each other.
实施例1，参考图1和图2，一种光学投影层析成像的装置，包括：平行光源器、相机5、远心成像系统4、步进电机5、旋转样品台10、计算机9，所述平行光源器由白色光源器1和扩散片2构成，所述计算机9与所述相机5电连，所述平行光源器、旋转样品台10、远心成像系统4、相机5依次通过光路连接，所述远心成像系统4包括：前、后透镜、光阑，所述光阑位于所述前、后透镜之间，并位于所述前透镜的后焦面上；所述旋转样品台10上设有充满水的玻璃池3。所述相机5为COMS相机。Embodiment 1, with reference to Fig. 1 and Fig. 2, a device for optical projection tomography, comprising: a parallel light source device, a camera 5, a telecentric imaging system 4, a stepping motor 5, a rotating sample stage 10, and a computer 9, the The parallel light source device is composed of a white light source device 1 and a diffusion sheet 2, the computer 9 is electrically connected to the camera 5, and the parallel light source device, the rotating sample stage 10, the telecentric imaging system 4, and the camera 5 are sequentially connected through an optical path. , the telecentric imaging system 4 includes: front and rear lenses, a diaphragm, the diaphragm is located between the front and rear lenses, and is located on the rear focal plane of the front lens; the rotating sample stage 10 A glass pool 3 full of water is provided on the top. The camera 5 is a CMOS camera.
当本发明的装置工作时，首先，白色光源器1发出白光经即扩散片2成为照度均匀的平行束白光，照射在生物样品8上，由于生物样品一般会采用浓度为1％的琼脂固定，为了减少光通过琼脂时的折射率，因此，将生物样品8放置入充满水的玻璃池3，进行折射率匹配，再置于旋转平台10上，所述远心成像系统4的前、后透镜都为10×的显微目镜，光阑位于前透镜的后焦面，其中，所述远心成像系统4的光阑的作用是增加光阑可以减小光锥的锥角及光锥和主光轴的夹角，增大成像深度及减小系统误差，而同时在光阑两边加入两个透镜的作用是前透镜距样品的距离较远，样品经前透镜的像为缩小的实像，添加后透镜可将经过前透镜的缩小的实像进行一定程度的放大作用。When the device of the present invention works, at first, the white light emitted by the white light source 1 passes through the diffuser 2 to become a parallel bundle of white light with uniform illumination, and irradiates on the biological sample 8. Since the biological sample is generally fixed with 1% agar, In order to reduce the refractive index when the light passes through the agar, therefore, the biological sample 8 is placed into the glass tank 3 filled with water, the refractive index is matched, and then placed on the rotating platform 10, the front and rear lenses of the telecentric imaging system 4 Both are 10× microscope eyepieces, and the diaphragm is located on the rear focal plane of the front lens, wherein the function of the diaphragm of the telecentric imaging system 4 is to increase the diaphragm to reduce the cone angle of the light cone and the distance between the light cone and the main lens. The included angle of the optical axis increases the imaging depth and reduces the system error, and the effect of adding two lenses on both sides of the diaphragm at the same time is that the distance between the front lens and the sample is relatively long, and the image of the sample passing through the front lens is a reduced real image. The rear lens can magnify the reduced real image passed through the front lens to a certain extent.
相机5用于采集到一系列的投影数字图像并传到计算机9。其中相机5的型号为Basler acA2000-340k(Base),镜头的景深范围为2mm，放大倍率为0.16，曝光时间为0.09。The camera 5 is used to collect a series of projected digital images and transmit them to the computer 9 . The model of camera 5 is Basler acA2000-340k (Base), the depth of field of the lens is 2mm, the magnification is 0.16, and the exposure time is 0.09.
步进电机6驱动相机5移动，使得生物样品8与远心成像系统4的距离改变，从而改变了景深的初始位置，每驱动一次，等到相对静止后，拍摄一张，最终得到同一个角度上多张不同焦面的照片，本实施例为5张，并利用拉普拉斯金字塔算法将同一个角度不同焦面的图像融合成长景深图片。The stepping motor 6 drives the camera 5 to move, so that the distance between the biological sample 8 and the telecentric imaging system 4 changes, thereby changing the initial position of the depth of field. Every time it is driven, it waits until it is relatively stationary, and then takes a picture, finally obtaining the same angle. A plurality of photos with different focal planes, 5 in this embodiment, and using the Laplacian pyramid algorithm to fuse images of different focal planes at the same angle to form a depth-of-field picture.
伺服电机7将驱动放着生物样品8的旋转平台10，旋转平台10每转动1.8度，进行上述的步骤采集同一个角度上的图像，旋转平台10一个旋转200次，360度，再由相机5采集图像并将得到的投影数字图像传到计算机9，共融合200幅长景深图片，这些图片将由计算机9利用滤波反投影算法进行重建处理，其中滤波反投影算法的滤波函数为S－L函数。在得到不同旋转角度下各个剖析层投影量后，经计算机9按照算法重建就可得样品各个层析图像,其重建结果可以从不同的角度进行观察及得到沿不同方向的虚拟切片，进而可得出整个样品的三维光吸收层析结构。再用amira软件合成三维立体图像。The servo motor 7 will drive the rotating platform 10 on which the biological sample 8 is placed. Every time the rotating platform 10 rotates 1.8 degrees, the above steps are performed to collect images at the same angle. The rotating platform 10 rotates 200 times, 360 degrees, and then the camera 5 Collect images and transmit the projected digital images to the computer 9 to fuse 200 long depth-of-field pictures. These pictures will be reconstructed by the computer 9 using the filtered back-projection algorithm, where the filter function of the filtered back-projection algorithm is the SL function. After obtaining the projection volume of each analysis layer under different rotation angles, each tomographic image of the sample can be obtained by computer 9 reconstruction according to the algorithm, and the reconstruction results can be observed from different angles and virtual slices along different directions can be obtained, and then can be obtained The three-dimensional light absorption tomographic structure of the whole sample was obtained. Then use amira software to synthesize three-dimensional images.
该方法和装置，通过对每一个焦面时通过采集不同焦点的照片，再通过拉普拉斯金字塔算法，从而得出长景深照片，再利用滤波反投影算法进行重建，使得图像所提取的信息充分，图像分辨率高。The method and device collect photos with different focal points for each focal plane, and then use the Laplacian pyramid algorithm to obtain a long depth of field photo, and then use the filter back projection algorithm to reconstruct, so that the information extracted from the image Full, high-resolution images.
其中，所述拉普拉斯金字塔算法的原理步骤为：Wherein, the principle steps of the Laplacian Pyramid Algorithm are:
步骤一、对每一源图像分别进行梯度塔形分建立图像的梯度金字塔。Step 1: Perform gradient pyramid analysis on each source image to establish a gradient pyramid of the image.
步骤二、对图像梯度金字塔的各分解层分别进行融合处理；不同的分解层、不同方向细节图像采用不同的融合算子进行融合处理，最终得到融合后图像的梯度金字塔。Step 2: Fusion processing is performed on each decomposition layer of the image gradient pyramid; different decomposition layers and different direction detail images are fused with different fusion operators, and finally the gradient pyramid of the fused image is obtained.
步骤三、对融合后所得梯度金字塔进行逆塔形变换(即进行图像重构)。Step 3: Perform inverse pyramid transformation (that is, perform image reconstruction) on the gradient pyramid obtained after fusion.
以上对本发明的较佳实施方式进行了具体说明，但本发明创造并不限于所述实施例，熟悉本领域的技术人员在不违背本发明精神的前提下还可做出种种的等同变型或替换，这些等同的变型或替换均包含在本申请权利要求所限定的范围内。The preferred embodiments of the present invention have been described in detail above, but the invention is not limited to the described embodiments, and those skilled in the art can also make various equivalent modifications or replacements without violating the spirit of the present invention. , these equivalent modifications or replacements are all included within the scope defined by the claims of the present application.
Claims (7)
- A kind of 1. method of optical projection tomography, it is characterised in that including step：Shoot the image of several different angle difference focal planes of biological sample；Using laplacian pyramid algorithm by same angle Spend the image co-registration growth depth of field picture of different focal planes；Using filter back-projection algorithm by the long depth of field of biological sample different angle Picture reconstruction.
- 2. the method for a kind of optical projection tomography according to claim 1, it is characterised in that the step is given birth to Different focal planes described in the image of several different angle difference focal planes of thing sample are 5 different focal planes.
- 3. the method for a kind of optical projection tomography according to claim 1 or 2, it is characterised in that in the shooting Before the image of several different angle difference focal planes of biological sample, the biological sample is placed in water-filled aquarium.
- 4. a kind of device of optical projection tomography, including：Source of parallel light device, camera, telecentric imaging system, its feature exist In, stepper motor, specimen rotating holder, computer are further included, the computer is electrically connected with the camera, the source of parallel light device, Specimen rotating holder, telecentric imaging system, camera pass sequentially through light path connection, and the stepper motor is used to drive the camera towards life Thing sample moves linearly, and the specimen rotating holder is used to drive the biological sample to rotate, and the camera is used in stepper motor Under drive, under the rotation of specimen rotating holder, the image of several different angle difference focal planes of shooting biological sample, the computer For obtaining the image of camera shooting, and laplacian pyramid algorithm is utilized by the image co-registration of same angle difference focal plane Growth depth of field picture, 3-D view is formed using filter back-projection algorithm by the long depth of field picture of biological sample different angle.
- A kind of 5. device of optical projection tomography according to claim 4, it is characterised in that：The specimen rotating holder It is equipped with water-filled aquarium.
- A kind of 6. device of optical projection tomography according to claim 5, it is characterised in that：The telecentric imaging system System includes：Forward and backward lens, diaphragm, the diaphragm are located at the back focal plane of the front lens between the forward and backward lens On.
- 7. according to a kind of device of optical projection tomography of claim 4-6 any one of them, it is characterised in that：The phase Machine is COMS cameras.
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