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Second harmonic generation (SHG) microscopy represents a very powerful tool for tissue characterization. Polarization-resolved SHG (PSHG) microscopy extends the potential of SHG, by exploiting the dependence of SHG signals on the polarization state of the excitation beam. Among others, this dependence translates to the fact that SHG images collected under different polarization configurations exhibit distinct characteristics in terms of content and appearance. These characteristics hold deep implications over image quality, as perceived by human observers or by image analysis methods custom designed to automatically extract a quality factor from digital images. Our work addresses this subject, by investigating how basic image properties and the outputs of no-reference image quality assessment methods correlate to human expert opinion in the case of PSHG micrographs. Our evaluation framework is based on SHG imaging of collagen-based ocular tissues under different linear and elliptical polarization states of the incident light.

A Study on Image Quality in Polarization-Resolved Second Harmonic Generation Microscopy