Graf Benedikt W, Boppart Stephen A
Biophotonics Imaging Laboratory, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Methods Mol Biol. 2010;591:211-27. doi: 10.1007/978-1-60761-404-3_13.
Three-dimensional (3D) cell cultures are important tools in cell biology research and tissue engineering because they more closely resemble the architectural microenvironment of natural tissue, compared to standard two-dimensional cultures. Microscopy techniques that function well for thin, optically transparent cultures, however, are poorly suited for imaging 3D cell cultures. Three-dimensional cultures may be thick and highly scattering, preventing light from penetrating without significant distortion. Techniques that can image thicker biological specimens at high resolution include confocal microscopy, multiphoton microscopy, and optical coherence tomography. In this chapter, these three imaging modalities are described and demonstrated in the assessment of functional and structural features of 3D chitosin scaffolds, 3D micro-topographic substrates from poly-dimethyl siloxane molds, and 3D Matrigel cultures. Using these techniques, dynamic changes to cells in 3D microenvironments can be non-destructively assessed repeatedly over time.
三维（3D）细胞培养是细胞生物学研究和组织工程中的重要工具，因为与标准的二维培养相比，它们更接近天然组织的结构微环境。然而，对于薄的、光学透明的培养物效果良好的显微镜技术，并不适合对3D细胞培养进行成像。三维培养物可能很厚且高度散射，导致光线在没有明显失真的情况下无法穿透。能够对较厚生物标本进行高分辨率成像的技术包括共聚焦显微镜、多光子显微镜和光学相干断层扫描。在本章中，将描述并展示这三种成像方式在评估3D壳聚糖支架、由聚二甲基硅氧烷模具制成的3D微地形底物以及3D基质胶培养物的功能和结构特征方面的应用。使用这些技术，可以随着时间的推移对3D微环境中的细胞动态变化进行反复无损评估。
