Institute of Applied Physics "Nello Carrara", National Research Council, Sesto Fiorentino, Italy.
Biophys J. 2012 Sep 19;103(6):1179-87. doi: 10.1016/j.bpj.2012.07.055.
The thermal transitions of fibrillar collagen are investigated with second-harmonic generation polarization anisotropy microscopy. Second-harmonic generation images and polarization anisotropy profiles of corneal stroma heated in the 35-80°C range are analyzed by means of a theoretical model that is suitable to probe principal intramolecular and interfibrillar parameters of immediate physiological interest. Our results depict the tissue modification with temperature as the interplay of three destructuration stages at different hierarchical levels of collagen assembly including its tertiary structure and interfibrillar alignment, thus supporting and extending previous findings. This method holds the promise of a quantitative inspection of fundamental biophysical and biochemical processes and may find future applications in real-time and postsurgical functional imaging of collagen-rich tissues subjected to thermal treatments.
采用二次谐波产生偏振各向异性显微镜研究了纤维状胶原蛋白的热转变。通过适用于探测即刻生理相关的主要分子内和纤维间参数的理论模型，分析了在 35-80°C 范围内加热的角膜基质的二次谐波产生图像和偏振各向异性分布。我们的结果描绘了随着温度的组织变化，这是包括其三级结构和纤维间排列的胶原组装的不同层次结构的三个去结构化阶段的相互作用，从而支持和扩展了以前的发现。这种方法有望对基本生物物理和生物化学过程进行定量检查，并可能在经受热治疗的富含胶原蛋白的组织的实时和手术后功能成像中找到未来的应用。
