Second harmonic generation imaging of fascia within thick tissue block

Comparing the SHG image formation for thin sections of tail tendon fascia and skeletal muscle fascia, we demonstrate that the forward (F) and backward (B) SHG images are vastly different. In addition, despite the different arrangement of the collagen Type I fibrillar architecture forming these two fascias, their ratios of forward over backward signal (F/B) are nearly equal. SHG images of thick tissue blocks of the fascia-muscle unit show only backward features, as opposed to SHG images of tissue blocks of the fascia-tendon unit. These images are an amalgamation of forward and backward features due to the backscattering of forward components within tendon. These forward features disappear when this tissue block is immersed in glycerol as backscattering is hereby suppressed.