大鼠早期急性心肌缺血中的荧光成像和光谱分析

采用结扎大鼠冠状动脉左前降支的方法建立急性心肌缺血模型,通过双光子荧光显微成像技术(TPEF)获取左心室前壁不同缺血时间的图象,得到心肌在不同缺血时间情况下的双光子荧光图像及其光谱图像。从荧光图像中得到不同时刻形态学信息及心肌细胞代谢信息;从光谱图像中分析心肌缺血组织物质成分。     

自适应光学在双光子显微成像技术中的应用

光学显微镜是生物医学研究必不可少的工具，其中双光子显微成像技术具有大深度三维显微成像功能，被认为是深层生物组织研究的首选工具。但是，在双光子成像系统使用过程中，光学系统的装配偏差、光学元件不理想以及生物样品的不均匀性都会在成像过程中引入像差，从而降低成像质量。通过在双光子显微成像系统中引入自适应光学技术，可实现对像差的有效校正，从而提高成像的分辨率、深度和视场。介绍了双光子显微成像中的像差来源和特点，概述了自适应光学技术中不同的探测和校正方法，综述了近年来自适应光学技术在双光子显微成像中不同的应用成果，最后对自适应光学在双光子显微成像中的发展进行了展望。     

罗丹明B的飞秒双光子荧光显微成像研究

利用宽带飞秒振荡器作为激发光源搭建了双光子荧光显微成像系统,在飞秒脉冲的宽带激发下,测量了罗丹明B溶液的双光子荧光光谱,开展了罗丹明B固体样品的双光子荧光显微成像研究。在双光子荧光显微成像研究中,通过调节激发脉冲的功率,分析了双光子荧光强度和激发脉冲功率之间的关系,并且利用半波片改变线偏振激发光场的偏振方向,研究了双光子荧光强度随激发脉冲偏振方向的变化趋势,实现了双光子荧光强度的类正弦调制。     

自适应光学技术在深层动态荧光显微成像中的应用和发展

荧光显微成像技术是开展微观生命科学研究的重要手段和工具,使用该技术可以观察生物体内的精细结构、动态追踪生物体内组织、细胞、细胞核、蛋白、小分子等不同尺度的生命活动过程。其中,研究深层组织高时空分辨率荧光显微成像技术,是当前成像领域一个前沿问题。应用自适应光学技术实时补偿经由不透明散射、非均匀生物组织传播而引入的复杂波前畸变已被证实是实现上述技术的一种有效途径。文中首先归纳了深层动态荧光显微成像的需求和特点,随后分别介绍了自适应光学技术近几年在共聚焦显微成像、随机光学重构显微成像、光激活定位显微成像、受激辐射光淬灭显微成像、双光子/多光子激发显微成像中的相关应用,并对今后的研究问题和发展方向提出展望。     

基于深度学习的荧光显微性能提升（特邀）

荧光显微镜具有对样品损伤小、可特异性成像等优点,是生物医学研究的主流成像手段。随着人工智能技术的快速发展,深度学习在逆问题求解中取得了巨大成功,被广泛应用于诸多领域。近年来,深度学习在荧光显微成像中的应用掀起了一个研究热潮,为荧光显微技术发展提供了性能上的突破与新思路。基于此,首先介绍了深度学习的基本网络模型,然后对基于深度学习的荧光显微成像技术在荧光显微的空间分辨率、图像采集及重建速度、成像通量和成像质量提升方面的应用进行阐述。最后,对目前深度学习在荧光显微成像中的研究进行总结与展望。     

利用波前调制技术提升光透明样品的双光子成像分辨率

组织光透明技术结合双光子显微成像(TPM)技术能够有效提升生物样品的显微成像深度,然而现有的光透明剂与常用显微物镜的浸润介质折射率并不匹配,会引入球差从而降低深层组织成像的荧光强度和分辨率。针对该问题分析了球差对TPM荧光强度和分辨率的影响,建立了由物镜特性(数值孔径和浸润介质)、聚焦深度和物体折射率等参数构成的球差补偿模型,进而指导空间光调制器进行球差补偿。对荧光小球仿体样品和光透明脑组织样品的双光子成像结果显示,该球差补偿方法能显著提升样品信号量和系统纵向分辨率。另外,该方法在校正过程中无需多次成像,操作简单且耗时短,对光透明剂和显微物镜无特殊要求,具有较强的通用性。     

微型化显微成像系统的关键技术及研究进展（特邀）

神经环路动态功能的解析是当前脑科学领域的重点和难点,微型化显微成像技术为其研究提供了重要手段。相较于双光子荧光成像和光纤光度法,微型化显微系统能够在模式动物自由活动状态下进行长时程、单细胞分辨率、实时成像。近十几年来,科学家们围绕可穿戴、高稳定性要求,先后研制了单光子、多光子成像系统,并从荧光探针、光电子元件、数据传输等方面进行不断优化,提升系统性能,扩展应用范围。将从成像原理、基本结构、系统优化、应用方案及未来发展方向等方面对微型化显微成像系统进行分析和讨论,综述各方向研究进展,旨在为该领域技术提升和神经科学应用提供参考。     

多光子成像中的生物组织光损伤

多光子激发荧光成像技术因低侵入性、强穿透力、高信噪比和高空间分辨率在生物医学光学领域得到广泛的应用，同时也成为最重要的研究工具之一。在多光子成像中过量的光子密度或激光功率会引起生物组织光损伤。光损伤决定了成像所能使用的激光功率的上限。光损伤强度与激光、组织光学参数有关，其背后的作用机制可分为光化学作用和光热作用。重点论述了光损伤的基本原理和形成机制，阐述了光损伤分析数学模型。讨论和分析了不同组织、不同波长下光损伤的一些研究进展。总结了光损伤规律：无色素组织双光子成像中光损伤以光化学作用为主，色素组织双光子成像中光损伤以光热作用为主，三光子深层组织成像中光损伤很可能来自光化学和光热协同作用。展望了降低光损伤和优化成像参数的可行策略。     

小鼠大脑飞秒双光子荧光三维显微成像研究

双光子荧光(two-photon fluorescence,TPF)显微成像技术借助荧光探针实现样品中被标记成分的特异性成像，具有天然的三维层析能力、高成像深度与空间分辨率、以及更小的光漂白与光损伤，已经发展成为化学、医药学和生命科学领域的一项重要研究工具。文中通过分析高斯光束复振幅在空间中的分布，推导出TPF信号的纵向与径向分布公式，以此估算出文中的TPF显微成像系统的横向分辨率为453 nm，纵向分辨率为2.087μm。使用飞秒激光器作为激发光源，搭建了TPF显微成像系统。在800 nm波长的飞秒脉冲激发下，测量了罗丹明B溶液的TPF光谱，从而选择636～703 nm作为显微成像的荧光探测窗口。随后开展了对罗丹明B染色的小鼠大脑切片的TPF显微成像实验研究，利用断层扫描成像的方式获得了小鼠大脑切片在0～14μm深度内的荧光强度分布。通过三维重构完成了对生物样品的三维立体成像，获得了小鼠大脑中灰质与白质在不同深度的分布情况，实验结果证明了搭建的显微成像系统具有优异的成像深度与空间分辨能力。     

甲状腺组织的双光子荧光成像

分别对离体新鲜的正常甲状腺组织、结节性甲状腺肿组织以及甲状腺乳头状癌组织进行自体双光子激发荧光(TPEF)和二次谐波(SHG)成像的研究.从双光子图像可知,正常甲状腺组织中滤泡大小较均匀,形态较相似,结节性甲状腺肿滤泡大小不等,而甲状腺癌组织则为大量的实质性癌细胞团结构,结果与标准组织学苏木精一伊红(H＆E)染色图相一致.此外,正常甲状腺组织和结节性甲状腺肿在胶原分布上存在一定差别.研究表明,双光子荧光成像技术可在微观结构上分辨出正常甲状腺组织、结节性甲状腺肿组织以及甲状腺乳头状癌组织的形态学差异,并有望成为甲状腺微创、快速诊断的有效方法.     

Spectral imaging technology of epithelial tissue based on two-photon excited fluorescence and second-harmonic generation

The layer structures of the esophageal and oral tissues were investigated by using spectral imaging technology based on two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG). Because spectral imaging technology allows a simultaneous record of both the spectra and image, it is capable of identifying the layered structures of the epithelial tissues, including the keratinizing layer, epithelial cell layer and stromal layer in the molecular level, which are strongly correlated to tissue pathology. All this work indicates that this technique has the potential to provide more accurate and comprehensive information for the early pathological diagnosis of tissues with the stratified squamous epithelia.     

Molecular Engineering of NIR-II AIE Luminogen Excited at 1700 nm for Ultradeep Intravital Brain Two-Photon Fluorescence Imaging

The limited tissue penetration depth and spatial resolution are the major bottlenecks for deep-brain imaging. In this study, molecular engineering by tailoring electron donors is conducted to develop for the first time an NIR-II (second near-infrared) emissive fluorescence probe, namely DCTBT, for effective deep-brain two-photon fluorescence imaging. Benefiting from its good biocompatibility, high photostability, bright NIR-II emission as aggregates and large two-photon fluorescence action cross section at the 1700 nm excitation window, DCTBT offers the imaging depths of 2180 and 1135 µm in mouse brain with removed and intact skull, respectively. These results are the record depths for brain imaging, compared to all kinds of fluorescent probes and all modalities of multiphoton microscopy at all demonstrated excitation wavelengths. Moreover, with DCTBT labeling, hemodynamic imaging of blood flow in mouse brain vessels down to a depth of 714 µm with the intact skull is achieved. Multiphoton fluorescence imaging with the NIR-II probe DCTBT excited at the 1700 nm window may readily provide methodology for deep-brain structural and hemodynamic research.     

Dielectrical spectroscopy of canine myocardium during acute ischemia and hypoxia at frequency spectrum from 100 kHz to 6 GHz

We studied dielectrical properties of canine myocardium during acute ischemia and hypoxia using dielectrical spectroscopy method at frequency spectrum from 100 kHz to 6 GHz. This study was conducted on a group of six canines with acute ischemia and seven canines with hypoxia. Hypoxia (10% for 30 min) decreases myocardial resistance (rho), while the dielectrical permittivity (epsilon') of the myocardial tissue remains statistically unchanged. Acute ischemia for 2 hr causes significant frequency-dependent changes in both epsilon' and rho of myocardial tissue. Myocardial resistance increases, while the sign and amplitude of changes in the myocardial epsilon' are frequency and time dependent. These observations open up an opportunity for assessing the properties of myocardial tissue using dielectrical spectroscopy as well as noninvasively with the help of imaging methods based on electrical impedance and microwave tomography.     

Three-dimensional mapping of acute ischemic regions usingartificial neural networks and tagged MRI

Many methods for mapping ischemic myocardial regions by functional analysis have been suggested. However, the complicated relationship between myocardial function and perfusion, and the inherent limitations of the imaging techniques used, have led to a generally low mapping accuracy. The authors show herein, that highly accurate mapping can be obtained by combining tagged magnetic resonance imaging (MRI), three-dimensional (3-D) analysis, and artificial neural networks. Nine canine hearts with acute ischemia were studied using multiplanar tagged MRI. Twenty-four myocardial cuboids were tagged in each heart and reconstructed in 3-D at end diastole (ED) and end systole (ES). The cuboids were arranged in three slices approximately 1 cm thick and covered most of the left ventricle (LV). Transmural thickening and endocardial area strain were calculated for each cuboid. Applying a post-mortem (PM) analysis, the percent ischemia in each cuboid was estimated using monastral blue dye; the PM analysis served as a “gold standard”. An artificial neural network (ANN), designed to estimate the percent ischemia in each cuboid from the functional indexes, was then created. The ANN “learned” the function-ischemia relationship in 192 cuboids taken from eight of the hearts and was asked to estimate the percent ischemia in the 24 cuboids of the ninth heart. The process was repeated nine times, each time using a different heart as test case. The average accuracy of mapping, i.e., the accuracy with which the ANN has mapped the normal and ischemic cuboids using the functional parameters, was 87.5%±7.8 (s.d.). This accuracy was superior to the accuracy obtained by optimal thresholding of the same thickening (80.1%) and endocardial strain (76.9%) data     

基于THz-TDS的碳纤维复合材料无损检测

基于反射式THz-TDS成像技术对碳纤维缠绕增强复合材料缺陷进行无损检测实验,获得不同缺陷碳纤维样品的成像结果及数据。结果表明,反射式THz-TDS成像技术在0.1～3.5 THz波段对碳纤维复合材料中的热损伤、划伤缺陷、磨损缺陷及孔洞缺陷成像清晰,分辨率较高;且获得的时域波形对样品热损伤缺陷敏感,适用于局部检测对整体性能的判断。     

Time-resolved two-photon excitation fluorescence spectroscopy and microscopy using a high repetition rate streak camera

We present a time-resolved two-photon excitation fluorescence spectroscopy and a simultaneous time- and spectrum- resolved multifocal multiphoton microscopy system that is based on a high repetition rate picosecond streak camera for providing time- and spectrum- resolved measurement and imaging in biomedicine. The performance of the system is tested and characterized by the fluorescence spectrum and lifetime analysis of several standard fluorescent dyes and their mixtures. Spectrum-resolved fluorescence lifetime images of fluorescence beads are obtained. Potential applications of the system include clinical diagnostics and cell biology etc.     

单分子荧光成像技术下的运动员疲劳程度检测

无接触环境下对运动员疲劳程度进行检测是一项新型技术,提出基于单分子荧光成像技术的运动员疲劳程度检测方法,通过单分子荧光成像装置,获取运动员身体机能荧光信号,并计算荧光寿命数值,获取运动员心率、血氧饱和度、体温、运动能耗在不同运动强度下的变动值.将这4种身体机能荧光寿命变动值参数,带入BP神经网络模型进行迭代训练检测,获...     

飞秒激光HpD双光子荧光法诊断癌症

提出了飞秒激光外源性血卟啉 Hp D 双光子荧光诊断癌症的新方法 .采用自锁模掺钛蓝宝石激光器输出的飞秒激光脉冲激发注入 Hp D的小鼠癌组织部位 ,观察到 Hp D的双光子荧光谱 ,而对照的正常组织部位则观察不到明显的双光子荧光 .实验结果显示 ,该法有可能成为癌症诊断的有效方法     

小型高光谱图谱仪与激光雷达及其海洋应用

海洋是地球生态环境的重要一环,但人类对海洋资源的勘探和开采容易对其造成严重破坏,如油气开采过程造成的大面积溢油、污染和赤潮爆发等。高光谱成像技术可以同时获取图像信息与高分辨光谱信息,在海洋原位探测上具有重大应用。文中综述了小型高光谱图谱仪与激光雷达及其在海洋应用上的部分近期工作。小型高光谱图谱仪结合荧光技术,实现了溢油种类的分类和油膜厚度的估计。多模式高光谱海洋原位探测系统可以工作于普通反射或透射成像、望远成像、显微成像三种模式,实现了海洋不同藻类及鱼类传染病载体孢囊的高光谱探测。高光谱技术结合激光雷达技术在溢油、赤潮等海洋污染物监测方面具有很大潜力。非弹性高光谱沙姆激光雷达系统通过油品的荧光光谱实现了海洋溢油油品的遥测鉴别。形貌沙姆激光雷达系统基于二维沙姆成像原理,通过空气-水界面折射矫正,成功的对人体、贝壳、珊瑚等进行了三维形貌重构,近处恢复精度可达毫米级,表面纹路清晰可见,为海洋监测应用提供了新的技术支持。