Department of Materials, Imperial College London, London SW7 2AZ, UK.
Department of Bioengineering, Imperial College London, London SW7 2AZ, UK.
Nat Commun. 2017 Mar 22;8:14843. doi: 10.1038/ncomms14843.
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.
同时对生物相关的三维（3D）细胞培养环境中的多个生物分子进行成像的能力，将极大地促进对复杂细胞机制和细胞-材料相互作用的理解。在这里，我们提出了一种用于无标记定量体积拉曼成像（qVRI）的计算框架。我们将 qVRI 应用于一系列生物系统：具有心脏衍生物的人类多能干细胞、传统细胞培养系统中的单核细胞和单核细胞衍生的巨噬细胞以及仿生水凝胶内的间充质干细胞，这些水凝胶为 3D 细胞培养环境提供了支持。我们展示了使用振动微光谱术对细胞形状、细胞质、细胞核、脂滴和细胞骨架结构进行三维可视化和定量分析，具有前所未有的生物分子特异性。
