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. 2007 Apr 20;93(4):1312–1320. doi: 10.1529/biophysj.106.100586

FIGURE 2.

FIGURE 2

SHG images and ratio measurements for Achilles tendon. (a–c) Approximately 10 μm sample thickness. (a) Forward image. (b) Backward image. (c) 20% Intralipid emulsion added onto the thin sample section. (d) Image of thick intact, ex vivo tissue block (∼5 mm thickness). All images have 1024 × 1024 pixels and the dimension is x = 67 μm and y = 67 μm. The image of the intact, thick tendon tissue (d) strongly resembles the image obtained with intralipid emulsion on top of a thin sample section (c), suggesting that backscattering is important for Achilles tendon imaging. Ratio measurements are (e) F/B, (f) LE/B, and (g) T/B. These ratios are outlined in Experimental Methods. The average ratios are F/B=24.9±2.4, LE/B=10.6±1.4, and T/B=6.2±0.8. These ratios are used to determine the fraction of forward signal contributing to the overall backward signal due to backscattering. For whole, thick ex vivo tendon, Pbs = 21 ± 6%. Adding a 20% intralipid emulsion on top of the thin sample section, Pbs = 39 ± 10%.