无标记定量共聚焦位相显微技术

项目名称： 无标记定量共聚焦位相显微技术

项目编号： No.61475010

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 无线电电子学、电信技术

项目作者： 席鹏

作者单位： 北京大学

项目金额： 80万元

中文摘要： 定量位相探测在生命科学研究中具有重要意义。虽然共聚焦显微技术已经在细胞研究中得到广泛应用，但是它无法直接获取细胞位相信息。利用差分位相干涉可以获取位相信息，但其光强-位相关系为非线性；同时它需要利用二维探测器，无法直接与共聚焦技术兼容。我们通过激光斜射扫描显微技术(LOSOM)，成功将共聚焦荧光成像与位相成像相结合。在本申请中，我们提出一种基于LOSOM的定量位相成像技术。利用Schlieren共聚焦部分遮挡能够获取位相一阶偏导数的特点，我们设计了两种实现位相成像的方案：1）利用XY轴的快门光阑实现四个方向的偏导数测量；2）利用一个载有四种不同发光光谱的量子点荧光编码板实现四个方向的偏导数测量。然后，通过求解线性方程实现反推位相。这一工作能够充分利用LOSOM技术与位相导数直接相关的特点，实现LOSOM从定性到定量成像的进展。本工作将实现定量位相与定量荧光双模态成像。

中文关键词： 共聚焦；位相；斜射照明；无标记

英文摘要： Quantitative phase detection is of important significance in life science. Although confocal microscopy has been widely used in cellular study, it cannot obtain the phase information of the specimen directly. Through differential contrast interference (DIC), phase information can be extracted. Yet, DIC has a nonlinear relationship between the intensity and the phase. Also, it requires two-dimensional detector, which is incompatible for confocal microscopy. By using laser oblique scanning optical microscopy (LOSOM), we have successfully combined confocal fluorescence microscopy with phase imaging. In this application, we proposed a novel quantitative phase imaging method based on LOSOM. Taking advantage of the linear relationship between the phase derivative and the intensity in Schilieren partial-obstruction microscopy, we designed two schemes: 1) Using a dual-shutter obstructer in XY axes, the four local derivatives can be collected; 2) Using a fluorescent plate loaded with four different kinds of quantum dots, the four local derivatives can be obtained by spectral collection. Then, through solving the linear equation to obtain the phase of the specimen. This work can push LOSOM from qualitative phase microscopy to quantitative phase microscopy, taking advantage of the linear relationship between the phase and the intensity in LOSOM. As LOSOM is highly compatible with confocal microscopy, the proposed method can achieve dual modality imaging combining both quantitative phase microscopy and quantitative fluorescence microscopy.

英文关键词： confocal;phase;oblique illumination;label free