In situ three-dimensional monitoring of collagen fibrillogenesis using SHG microscopy.
In situ three-dimensional monitoring of collagen fibrillogenesis using SHG microscopy.
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DOI：
10.1364/boe.3.001446
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发表时间：
2012-06-01
影响因子：
3.4
通讯作者：
Schanne-Klein MC
中科院分区：
文献类型：
--
作者：
Bancelin S;Aimé C;Coradin T;Schanne-Klein MC
我们实施了原位延时二次谐波发生（SHG）显微镜监测的三维（3D）自组装胶原蛋白在溶液中。作为概念的证明，我们通过改变pH来调整原纤维形成的动力学，并测量了随后在SHG图像中原纤维体积密度的指数增加。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，我们可以专注于一个单一的分离的胶原纤维的生长，因为SHG显微镜是敏感的组织良好的纤维直径低于光学分辨率。这项工作说明了潜在的SHG显微镜的合理设计和表征的胶原蛋白为基础的生物材料。
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.