High resolution imaging of collagen organisation and synthesis using a versatile collagen specific probe
High resolution imaging of collagen organisation and synthesis using a versatile collagen specific probe
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DOI：
10.1016/j.jsb.2007.04.008
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发表时间：
2007-09-01
影响因子：
3
通讯作者：
Bouten, Carlijn V. C.
中科院分区：
文献类型：
--
作者：
Boerboom, Ralf A.;Krahn, Katy Nash;Bouten, Carlijn V. C.
胶原蛋白是主要负责组织的承载性质的蛋白质，并且胶原蛋白结构是组织的机械性质的主要决定因素之一。为了提高我们对胶原纤维形成和重塑的理解，例如在组织工程实验中，胶原三维结构变化的可视化是必不可少的。最近开发的胶原蛋白探针，基于天然胶原蛋白结合蛋白（CNA35）共轭的荧光染料，显示出更具体的胶原蛋白比现有的荧光技术目前用于胶原蛋白可视化活组织。在本文中，成像与荧光CNA35探针进行了比较，成像与二次谐波产生（SHG）和成像的二维和三维胶原组织进一步发展。对一系列样本（细胞培养物、血管和工程组织）进行成像，以说明这种胶原蛋白探针的潜力。与SHG生成的图像相比，胶原组织的图像显示出改善的细节，SHG是目前观察组织中三维胶原组织的最有效方法。总之，荧光CNA35探针可以轻松获得胶原蛋白的高分辨率成像，从非常年轻的原纤维到更成熟的胶原蛋白纤维。此外，这种探针能够实时可视化细胞培养中的胶原蛋白合成，这为研究胶原蛋白合成和重塑提供了新的机会。(c)2007年爱思唯尔公司All rights reserved.
Collagen is the protein primarily responsible for the load-bearing properties of tissues and collagen architecture is one of the main determinants of the mechanical properties of tissues. Visualisation of changes in collagen three-dimensional structure is essential in order to improve our understanding of collagen fibril formation and remodelling, e.g. in tissue engineering experiments. A recently developed collagen probe, based on a natural collagen binding protein (CNA35) conjugated to a fluorescent dye, showed to be much more specific to collagen than existing fluorescent techniques currently used for collagen visualisation in live tissues. In this paper, imaging with this fluorescent CNA35 probe was compared to imaging with second harmonic generation (SHG) and the imaging of two- and three-dimensional collagen organisation was further developed. A range of samples (cell culture, blood vessels and engineered tissues) was imaged to illustrate the potential of this collagen probe. This images of collagen organisation showed improved detail compared to images generated with SHG, which is currently the most effective method for viewing three-dimensional collagen organisation in tissues. In conclusion, the fluorescent CNA35 probe allows easy access to high resolution imaging of collagen, ranging from very young fibrils to more mature collagen fibres. Furthermore, this probe enabled real-time visualisation of collagen synthesis in cell culture, which provides new opportunities to study collagen synthesis and remodelling. (c) 2007 Elsevier Inc. All rights reserved.