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Collagen imaging using two-photon-generated second-harmonic generation (SHG)
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Two days after the last administration of treatment, collagen deposition was observed using SHG microscopy at 840 nm on a custom-designed, video-rate laser-scanning microscope. We randomly selected five areas of tumor tissue and acquired a short video of SHG signals. We then averaged the noise over 30 frames using MATLAB to improve contrast and signal-to-noise ratio. The positive signal area was analyzed by a blinded observer using Image J software.
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