Mandracchia Biagio, Gennari Oriella, Marchesano Valentina, Paturzo Melania, Ferraro Pietro
CNR - ISASI Institute of Applied Sciences and Intelligent Systems, Via Campi Flegrei 34, 80078, Pozzuoli, Napoli, Italy.
Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale - DICMAPI, University of Naples Federico II, Piazzale Tecchio 80, 80100, Napoli, Italy.
J Biophotonics. 2017 Sep;10(9):1163-1170. doi: 10.1002/jbio.201600177. Epub 2016 Nov 2.
The study of cell adhesion contacts is pivotal to understand cell mechanics and interaction at substrates or chemical and physical stimuli. We designed and built a HoloTIR microscope for label-free quantitative phase imaging of total internal reflection. Here we show for the first time that HoloTIR is a good choice for label-free study of focal contacts and of cell/substrate interaction as its sensitivity is enhanced in comparison with standard TIR microscopy. Finally, the simplicity of implementation and relative low cost, due to the requirement of less optical components, make HoloTIR a reasonable alternative, or even an addition, to TIRF microscopy for mapping cell/substratum topography. As a proof of concept, we studied the formation of focal contacts of fibroblasts on three substrates with different levels of affinity for cell adhesion.
细胞黏附接触的研究对于理解细胞力学以及细胞在底物上的相互作用或对化学和物理刺激的反应至关重要。我们设计并构建了一台用于全内反射无标记定量相位成像的全息全内反射显微镜（HoloTIR显微镜）。在此我们首次表明，全息全内反射显微镜是用于无标记研究黏着斑和细胞/底物相互作用的良好选择，因为与标准全内反射显微镜相比，其灵敏度有所提高。最后，由于所需光学组件较少，全息全内反射显微镜的实施简单且成本相对较低，这使其成为全内反射荧光显微镜（TIRF显微镜）用于绘制细胞/基质形貌的合理替代方案，甚至可以作为补充。作为概念验证，我们研究了成纤维细胞在三种对细胞黏附具有不同亲和力水平的底物上黏着斑的形成。
