Bancelin S, Aimé C, Coradin T, Schanne-Klein M-C
Biomed Opt Express. 2012 Jun 1;3(6):1446-54. doi: 10.1364/BOE.3.001446. Epub 2012 May 21.
We implemented in situ time-lapse Second Harmonic Generation (SHG) microscopy to monitor the three-dimensional (3D) self-assembly of collagen in solution. As a proof of concept, we tuned the kinetics of fibril formation by varying the pH and measured the subsequent exponential increase of fibril volume density in SHG images. We obtained significantly different time constants at pH = 6.5 ± 0.3 and at pH = 7.5 ± 0.3. Moreover, we showed that we could focus on the growth of a single isolated collagen fibril because SHG microscopy is sensitive to well-organized fibrils with diameter below the optical resolution. This work illustrates the potential of SHG microscopy for the rational design and characterization of collagen-based biomaterials.
我们采用原位延时二次谐波产生（SHG）显微镜来监测溶液中胶原蛋白的三维（3D）自组装过程。作为概念验证，我们通过改变pH值来调节原纤维形成的动力学，并在SHG图像中测量随后原纤维体积密度的指数增长。我们在pH = 6.5 ± 0.3和pH = 7.5 ± 0.3时获得了显著不同的时间常数。此外，我们表明可以聚焦于单个孤立胶原蛋白原纤维的生长，因为SHG显微镜对直径低于光学分辨率的有序原纤维敏感。这项工作说明了SHG显微镜在基于胶原蛋白的生物材料的合理设计和表征方面的潜力。
