早期胚胎全场OCT图像的边界去模糊研究

全场光学相干层析成像技术(全场OCT)是研究早期胚胎形态发育的最理想成像设备,然而所采集图像难免受噪声干扰.这些噪声可模糊早期胚胎内不同组织结构的边界,从而给基于图像边界的结构划分带来干扰.为解决这一问题,本文运用中值滤波、维纳滤波、各向异性扩散算法处理全场OCT获得的早期胚胎图像,并运用信噪比、均方误差、峰值信噪比和边缘保留等指标评价图像处理效果.结果表明:经各向异性扩散算法处理的早期胚胎图像,可完整地保留原始图像信息,且边界最清晰,视觉效果最好.     

高动态光学血管造影成像

我们提出一种高动态光学血管造影成像(HDOA)方法来实现活体生物样本血管造影成像.该方法通过设置高动态范围曝光时间,依据动态积分效应和吸收效应以实现高动态积分时间调制.通过该方法,不仅能够同时获得各级血管清晰的造影图像,还能消除样品厚度不均、吸收系数不同对成像造成的影响.论文以仿体和活体金鱼为样品,通过实验验证了HDOA方法根据动态积分调制效应和吸收效应,能有效实现各级血管同时成像.     

高动态光学血管造影成像（英文）

我们提出一种高动态光学血管造影成像(HDOA)方法来实现活体生物样本血管造影成像.该方法通过设置高动态范围曝光时间,依据动态积分效应和吸收效应以实现高动态积分时间调制.通过该方法,不仅能够同时获得各级血管清晰的造影图像,还能消除样品厚度不均、吸收系数不同对成像造成的影响.论文以仿体和活体金鱼为样品,通过实验验证了HDOA方法根据动态积分调制效应和吸收效应,能有效实现各级血管同时成像.     

基于空间频域滤波高动态光学投影层析的斑马鱼三维成像研究

本文提出了一种基于空间频率滤波的多曝光融合的高动态投影层析三维成像方法,实现了活体斑马鱼（17 mm × 4 mm,最大厚度为2.33 mm,最小厚度为0.29 mm）的三维结构成像. 通过相机采用不同曝光时间记录系列吸收图像,将每张图像取变换到频域去除低频后,将各张滤波后叠加并逆傅里叶变换回空域,对变换后的图像进行归一化处理,最终获得高动态图像. 在每个投影角度获得这种高动态吸收投影图像,进行滤波反投影算法重建,获得高动态的整条斑马鱼三维结构信息. 实验成像结果表明,这种空间频率滤波多曝光融合的高动态光学投影层析三维成像研究,可以获得复杂结构更丰富的空间信息,对斑马鱼等模式生物早期胚胎生长发育进程进行监测和定量评估有一定的应用前景.     

基于主成分分析的双对比光学投影断层成像

目的 本文提出了一种基于主成分分析（PCA）的双对比光学投影断层成像（DC-OPT）方法,以获得活体中血流网络和骨骼的三维可视化。方法 使用主成分分析方法来提取吸收图像和血流图像,原始图像序列的第一主成分用于获取吸收图像;通过计算每个像素的调制深度来获得流动图像。不同投影位置的流动和吸收对比图像被用于三维血流网络和骨骼的同步重建。结果 采用PCA和OPT相结合的方法,通过将动态血流信号和静态背景信号分离,实现了对微生物样本的血流网络和骨骼的三维成像。结论 本文研究的新颖之处在于通过同一光学系统获得了快速、同步、双对比的血流网络和骨骼三维图像。实验结果可用于活体生物的生理发育研究。     

金鱼雌核发育单倍体胚胎血液循环障碍产生机制

为研究鱼类单倍体血液循环障碍产生机制,人工诱导获得金鱼(Carassius auratus)雌核发育单倍体胚胎并进行活体观察及邻联茴香胺染色,结果显示金鱼雌核发育单倍体胚胎存在不同程度的血液循环不良和红细胞生成缺陷.为进一步探讨其发生的分子机制,利用反义RNA整胚原位杂交技术比较分析了原始造血和血管发生关键基因scl(...     

小动物活体光学三维成像系统及其对乳腺癌的定量分析

乳腺癌具有高转移率。使用小动物活体成像技术对乳腺癌的生长及转移情况实时监测定量分析可以帮助了解疾病机制及进行药物研究。二维成像对光学信号的定位与定量是相对的。随着计算机技术的进步,可以实现对采集的图像进行三维重建,精准量化光学信号,获得空间分布的三维信息。IVIS Spectrum小动物活体光学三维成像系统同时具有高灵敏的生物发光、荧光、切伦科夫辐射二维成像及三维扫描重建功能,是小动物活体光学成像的顶级系统。本文对人乳腺癌细胞（MDA-MB-231）进行慢病毒感染,在体外稳定表达荧光素酶后,选取重度联合免疫缺陷（SCID）小鼠进行原位乳腺癌模型的建立,通过IVIS Spectrum小动物活体光学三维成像系统对小鼠进行生物发光二维成像,无创观测肿瘤的生长及转移情况。本文的创新点是利用生物发光成像断层扫描技术对小鼠模型进行定量三维成像,使用系统自带的算法直接进行三维重建,同时结合鲸鱼优化算法（WOA）优化后的三维卷积的深度编码器-解码器的网络模型进行重建。通过CT图像验证两者的重建效果,得到肿瘤的深度信息,实现对乳腺癌的精准定量分析。     

Balance-FFE序列在胆管磁共振检查中的应用

目的:探讨磁共振平衡式稳态自由进动梯度回波序列(Balance-FFE)在胆管疾病中的应用价值.方法:92例胆管病变患者均进行了冠状面的Balance-FFE序列扫描和磁共振胰胆管造影(MRCP).将Balance-FFE的图像和3D MRCP像及MRCP原始图像对病变的显示率进行x2检验.结果:胆管系统在Balance-FFE序列中呈明显高信号,并能直接显示胆管结石和胆管狭窄,还能显示胆管外病变和胰腺病变,对周围淋巴结的显示也比较清楚.经x2检验,Balance-FFE序列和MRCP原始像对胆管系统病变的显示没有统计学意义(P0.05),而3DMRCP像对病变的显示能力不如Balance-FFE序列和MRCP原始像,对胆道病变的显示能力具有统计学意义(P<0.05).结论:Balance-FFE序列对胆道病变能清晰显示,成像速度快,图像信噪比高,伪影较少,与MRCP结合能提高对胆管系统病变的诊断率,因此Balance-FFE序列应作为胆管疾病磁共振扫描的常规序列,可作为MRCP序列的有效补充.     

基于压缩感知和散斑相关法的散射介质成像方法研究

散射介质成像是生物医学成像领域的一个重要研究方向,对生物医学临床的诊断有着重大的意义。结合散斑相关法和压缩感知技术,提出了一种散射介质成像方法。该方法与传统散射介质成像方法相比,将减少图像采集、图像重建所记录的数据量,提高图像处理的效率,并且降低了系统的搭建成本。试验结果表明,结合TVAL3信号重构算法和双谱分析法的散射图像重建算法,随着采样率的增大,峰值信噪比平稳上升,为散射介质成像方法在生物医学成像领域的运用提供了一种有效方案。     

基于光谱编码的血管内光声组分成像

光声成像突破了传统的光学成像和超声成像在生物组织成像领域的困境,该技术基于光声(Photoacoustic,PA)效应,脉冲激光激励下的生物组织产生超声信号,超声信号被接收后,通过反投影算法将其携带的时间信息和强度信息转化为能够反映生物组织吸收结构和分布的可视化图像。基于不同生物组织的光吸收差异,当激发光强度均匀且稳定时,光声成像反映的就是该物质对于该波长光的吸收特性。本文中,我们基于导管式的血管内光声断层扫描平台结合多波长激发的光声成像算法开发了基于光谱编码的血管内光声组分成像系统,实现了在离体血管斑块中脂质组分的定量成像,高分辨获得了脂质核心的大小形态和边界信息,表征了斑块内的脂质相对含量。     

Large‐depth‐of‐field full‐field optical angiography

A large‐depth‐of‐field full‐field optical angiography (LD‐FFOA) method is developed to expand the depth‐of‐field (DOF) using a contrast pyramid fusion algorithm (CPFA). The absorption intensity fluctuation modulation effect is utilized to obtain full‐field optical angiography (FFOA) images at different focus positions. The CPFA is used to process these FFOA images with different focuses. By selecting high‐contrast areas, the CPFA can highlight the characteristics and details of blood vessels to obtain LD‐FFOA images. In the optimal case of the proposed method, the DOF for FFOA is more than tripled using 10 differently focused FFOA images. Both the phantom and animal experimental results show that the LD‐FFOA resolves FFOA defocusing issues induced by surface and thickness inhomogeneities in biological samples. The proposed method can be potentially applied to practical biological experiments.      

基于Renyi熵滤波的光声图像重建算法设计与实现

针对光声图像重建过程中存在的原始光声信号信噪比差、重建图像对比度低、分辨率不足等问题,提出了基于Renyi熵的光声图像重建滤波算法.该算法首先根据原始光声信号的Renyi熵分布情况,确定分割阈值,并滤除杂波信号;再利用滤波后的光声数据进行延时叠加光声图像重建.利用该滤波算法分别处理铅笔芯横截面(零维)、头发丝(一维)以及小鼠大脑皮层血管(二维)等不同维度样本的光声信号,实验结果表明:相比Renyi熵处理之前,重建图像对比度平均增强了32.45%,分辨率平均提高了30.78%,信噪比提高了47.66%,均方误差降低了35.01%;相比典型的滤波处理算法(模极大值法和阈值去噪法),本研究中图像的对比度、分辨率和信噪比分别提高了25.94%/10.60%、27.90%/19.48%、35.21%/10.60%,均方误差减小了28.57%/16.66%.因此,选择利用Renyi熵滤波算法处理光声信号,从而使光声图像重建质量得到大幅改善.     

In vivo full‐field functional optical hemocytometer

We present an in vivo lab‐free full‐field functional optical hemocytometer (FFOH) for application to the capillaries of a live biological specimen, based on the absorption intensity fluctuation modulation (AIFM) effect. Because of the absorption difference between the red blood cells (RBCs) and background tissue under low‐coherence light illumination, an endogenous instantaneous intensity fluctuation is generated by the AIFM effect when RBCs discontinuously traverse the capillary. The AIFM effect is used to highlight the RBC signal relative to the background tissue by computing the real‐time modulation depth. FFOH can simultaneously provide a flow video, the flow velocity and the RBC count. Ourexperimental results can potentially be applied to study the physiological mechanisms of the blood circulation systems of near‐transparent live biological samples.      

Flow Measurements in a Blood-Perfused Collagen Vessel Using X-Ray Micro-Particle Image Velocimetry

Blood-perfused tissue models are joining the emerging field of tumor engineering because they provide new avenues for modulation of the tumor microenvironment and preclinical evaluation of the therapeutic potential of new treatments. The characterization of fluid flow parameters in such in-vitro perfused tissue models is a critical step towards better understanding and manipulating the tumor microenvironment. However, traditional optical flow measurement methods are inapplicable because of the opacity of blood and the thickness of the tissue sample. In order to overcome the limitations of optical method we demonstrate the feasibility of using phase-contrast x-ray imaging to perform microscale particle image velocimetry (PIV) measurements of flow in blood perfused hydrated tissue-representative microvessels. However, phase contrast x-ray images significantly depart from the traditional PIV image paradigm, as they have high intensity background, very low signal-to-noise ratio, and volume integration effects. Hence, in order to achieve accurate measurements special attention must be paid to the image processing and PIV cross-correlation methodologies. Therefore we develop and demonstrate a methodology that incorporates image preprocessing as well as advanced PIV cross-correlation methods to result in measured velocities within experimental uncertainty.     

Enhanced spatial resolution for snapshot hyperspectral imaging of blood perfusion and melanin information within human tissue

We report a reconstruction method to achieve high spatial resolution for hyperspectral imaging of chromophore features in skin in vivo. The method utilizes an established structure‐adaptive normalized convolution algorithm to reconstruct high spatial resolution of hyperspectral images from snapshot low‐resolution hyperspectral image sequences captured by a snapshot spectral camera. The reconstructed images at chromophore‐sensitive wavebands are used to map the skin features of interest. We demonstrate the method experimentally by mapping the blood perfusion and melanin features (moles) on the facial skin. The method relaxes the constrains of the relatively low spatial resolution in the snapshot hyperspectral camera, making it more usable in imaging applications.     

A Fast Image Alignment Approach for 2D Classification of Cryo-EM Images Using Spectral Clustering

Three-dimensional (3D) reconstruction in single-particle cryo-electron microscopy (cryo-EM) is a significant technique for recovering the 3D structure of proteins or other biological macromolecules from their two-dimensional (2D) noisy projection images taken from unknown random directions. Class averaging in single-particle cryo-EM is an important procedure for producing high-quality initial 3D structures, where image alignment is a fundamental step. In this paper, an efficient image alignment algorithm using 2D interpolation in the frequency domain of images is proposed to improve the estimation accuracy of alignment parameters of rotation angles and translational shifts between the two projection images, which can obtain subpixel and subangle accuracy. The proposed algorithm firstly uses the Fourier transform of two projection images to calculate a discrete cross-correlation matrix and then performs the 2D interpolation around the maximum value in the cross-correlation matrix. The alignment parameters are directly determined according to the position of the maximum value in the cross-correlation matrix after interpolation. Furthermore, the proposed image alignment algorithm and a spectral clustering algorithm are used to compute class averages for single-particle 3D reconstruction. The proposed image alignment algorithm is firstly tested on a Lena image and two cryo-EM datasets. Results show that the proposed image alignment algorithm can estimate the alignment parameters accurately and efficiently. The proposed method is also used to reconstruct preliminary 3D structures from a simulated cryo-EM dataset and a real cryo-EM dataset and to compare them with RELION. Experimental results show that the proposed method can obtain more high-quality class averages than RELION and can obtain higher reconstruction resolution than RELION even without iteration.     

Choriocapillaris and Choroidal Microvasculature Imaging with Ultrahigh Speed OCT Angiography

We demonstrate in vivo choriocapillaris and choroidal microvasculature imaging in normal human subjects using optical coherence tomography (OCT). An ultrahigh speed swept source OCT prototype at 1060 nm wavelengths with a 400 kHz A-scan rate is developed for three-dimensional ultrahigh speed imaging of the posterior eye. OCT angiography is used to image three-dimensional vascular structure without the need for exogenous fluorophores by detecting erythrocyte motion contrast between OCT intensity cross-sectional images acquired rapidly and repeatedly from the same location on the retina. En face OCT angiograms of the choriocapillaris and choroidal vasculature are visualized by acquiring cross-sectional OCT angiograms volumetrically via raster scanning and segmenting the three-dimensional angiographic data at multiple depths below the retinal pigment epithelium (RPE). Fine microvasculature of the choriocapillaris, as well as tightly packed networks of feeding arterioles and draining venules, can be visualized at different en face depths. Panoramic ultra-wide field stitched OCT angiograms of the choriocapillaris spanning ∼32 mm on the retina show distinct vascular structures at different fundus locations. Isolated smaller fields at the central fovea and ∼6 mm nasal to the fovea at the depths of the choriocapillaris and Sattler''s layer show vasculature structures consistent with established architectural morphology from histological and electron micrograph corrosion casting studies. Choriocapillaris imaging was performed in eight healthy volunteers with OCT angiograms successfully acquired from all subjects. These results demonstrate the feasibility of ultrahigh speed OCT for in vivo dye-free choriocapillaris and choroidal vasculature imaging, in addition to conventional structural imaging.