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We use second-harmonic generation (SHG) microscopy to quantitatively characterize collagen fiber crimping in the posterior cruciate ligament (PCL). The obtained SHG images are utilized to define three distinct categories of crimp organization in the PCL. Using our previously published spatial-frequency analysis, we develop a simple algorithm to quantitatively distinguish the various crimp patterns. In addition, SHG microscopy reveals both the three-dimensional structural variation in some PCL crimp patterns as well as an underlying helicity in these patterns that have mainly been observed using electron microscopy. Our work highlights how SHG microscopy could potentially be used to link the fibrous structural information in the PCL to its mechanical properties.

Application of quantitative second-harmonic generation microscopy to posterior cruciate ligament for crimp analysis studies