Abstract

Optical-based microscopy plays an important role in various scientific fields such as physics, chemistry and biology. Second harmonic generation (SHG) microscopy has become one of the indispensable tools for biomedical imaging for the last decade because the signal generated from SHG is sensitive to the objective structure and this amazing non-invasive method can also directly observe the objective without using extra fluorescent labels, especially for collagen molecules. As the most abundant protein in animals, collagen is responsible for a number of important structural and functional roles in vertebrates. For certain diseases, it has been shown that collagen fiber diameter has a significant variation and thus as a vital symptom for diagnosis. Moreover, collagen diameter is also a key parameter for fibrogenesis studying. Therefore, the determination of collagen fiber diameter is important for studying biophysical processes and identifying bioengineering applications. In this study, we investigated various collagen fibril thicknesses and the corresponding forward (FSHG) and backward (BSHG) second harmonic signal intensity variation. Our result exhibits that SHG intensity can quantify describe the relative collagen fibril thickness alteration, which also indicates the coherent effect difference between FSHG and BSHG. This approach demonstrates the capability of SHG imaging in providing collagen mechanical information and that may be applied in the evaluation of advancing collagen issues in vivo.

© 2011 OSA/SPIE