Vielreicher Martin, Friedrich Oliver
Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Institute of Medical Biotechnology, Paul-Gordan-Street 3, Erlangen, 91052, Germany.
Methods Mol Biol. 2017;1601:243-255. doi: 10.1007/978-1-4939-6960-9_19.
Multiphoton microscopy allows continuous depth-resolved, nondestructive imaging of scaffold-seeded cells during cell or tissue culture. Spectrally separated images in high resolution can be provided while cells are conserved in their native state. Here we describe the seeding of mesenchymal stem cells to bacterial nanocellulose hydropolymer scaffolds followed by 2-channel imaging of cellular autofluorescence (AF) and collagen-I formation using second harmonic generation (SHG) signals. With this approach the simultaneous observation of the progression of cell morphology and production of extracellular matrix as hallmarks of viability and cell fitness is possible.
多光子显微镜能够在细胞或组织培养过程中对接种于支架的细胞进行连续的深度分辨、无损成像。在细胞保持其天然状态的同时,可以提供高分辨率的光谱分离图像。在此,我们描述了将间充质干细胞接种到细菌纳米纤维素水凝胶支架上,随后利用二次谐波产生(SHG)信号对细胞自发荧光(AF)和I型胶原蛋白形成进行双通道成像。通过这种方法,可以同时观察细胞形态的变化以及细胞外基质的产生,这些都是细胞活力和健康状态的标志。