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We present a calibration protocol to obtain the alignment factors of a custom-made spectrometer and the nonlinear fitting function between the measured CCD pixel domain and the wavelength domain to apply to the spectral-domain optical coherence tomography (SD-OCT) using fiber Bragg gratings. We have used five gratings with different center wavelengths covering the broadband source spectral range. All have a narrow spectral bandwidth (0.05 nm) and the same reflectivity (92%) to calibrate and align the custom-made spectrometer. The implemented SD-OCT system following the proposed protocol showed the alignment factors as 44.37 deg incident angle, 53.11 deg diffraction angle, and 70.0-mm focal length. The spectral resolution of 0.187 nm was recalculated from the alignment factors.

We discuss the recent advances in the development and applications of second-order susceptibility as a contrast mechanism in optical microscopy for biological tissues. We review nonlinear optical methods and approaches for differentiation of tissue structures and discrimination of normal and pathological skin tissues, which have been demonstrated for the potential use in clinical diagnosis. In addition, the potential of second-order susceptibility imaging, encompassing applications in differentiating various types of collagen molecules for clinical diagnosis, is demonstrated. Finally, we discuss future development and application of this technique.

We present an automated and systematic two-dimensional discrete Fourier transform (2D-FFT) approach to analyze collagen fiber organization through the use of second harmonic generation (SHG) microscopy. Average orientations of individual domains and Ising-like order parameters introduced to characterize the correlation between orientations of adjacent domains may be used to quantitatively characterize fibrous tissues. Our approach was applied to analyze tissues including rat tail tendon, mouse skin, bovine corneas, and human corneas. We also show that collagen fiber organization in normal and keratokonus human corneas may be distinguished. The current approach may be used for the quantitative differentiation of SHG collagen fiber morphology in different tissues and may be applied for diagnostic purposes.