论文标题
三维双官能折射率和荧光显微镜(简介)
Three-dimensional bi-functional refractive index and fluorescence microscopy (BRIEF)
论文作者
论文摘要
荧光显微镜是成像具有分子特异性的生物样品的强大工具。相比之下,相显微镜提供了对样品折射率(RI)的无标签测量,该指数(RI)是一种固有的光学特性,与细胞形态,质量和刚度定量有关。传统的成像技术测量标记的荧光(功能)信息或无标签的RI(结构)信息,尽管具有两者可能很有价值。例如,生物组织具有异质性RI分布,导致样品诱导的散射降解荧光图像质量。当测量荧光和3D RI时,可以使用RI信息在荧光图像中数字纠正多散射效应。在这里,我们基于Epi-Mode显微镜开发了一种新的计算多模式成像方法,该方法可从单个数据集中重建3D荧光和3D RI。我们获取数十个荧光图像,每个图像被一个荧光团“照明”,然后通过多散射的正向模型解决一个逆问题。我们通过实验证明了我们的Epi-Mode 3D RI成像的方法和荧光图像中多散射效应的数字校正。
Fluorescence microscopy is a powerful tool for imaging biological samples with molecular specificity. In contrast, phase microscopy provides label-free measurement of the sample's refractive index (RI), which is an intrinsic optical property that quantitatively relates to cell morphology, mass, and stiffness. Conventional imaging techniques measure either the labeled fluorescence (functional) information or the label-free RI (structural) information, though it may be valuable to have both. For example, biological tissues have heterogeneous RI distributions, causing sample-induced scattering that degrades the fluorescence image quality. When both fluorescence and 3D RI are measured, one can use the RI information to digitally correct multiple-scattering effects in the fluorescence image. Here, we develop a new computational multi-modal imaging method based on epi-mode microscopy that reconstructs both 3D fluorescence and 3D RI from a single dataset. We acquire dozens of fluorescence images, each 'illuminated' by a single fluorophore, then solve an inverse problem with a multiple-scattering forward model. We experimentally demonstrate our method for epi-mode 3D RI imaging and digital correction of multiple-scattering effects in fluorescence images.