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S. Scaglione et al.:
Order versus Disorder: in vivo bone formation within osteoconductive scaffolds
Scientific Reports 2 (February 2012) 274, doi:10.1038/srep00274
Second harmonic generation (SHG) imaging microscopy was used to investigate the collagen matrix organization within bone tissue. A SHG and two-photon excitation fluorescence (2PEF) microscope was used for this study. The SHG/2PEF microscope was based on a Leica TCS STED CW, Resonant Scanner Spectral confocal scanning head (Leica Microsystems, Mannheim, Germany). The SHG is collected both in back-scattering (BSHG) and in a transmitted light configuration (forward second harmonic generation; FSHG). Since the excitation wavelength is set at 860 nm, the BSHG spectral component of interest, 415–445 nm, is selected by means of the Leica built-in prism based spectrophotometer, whereas the FSHG is selected by two filters, an