Abstract
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.
Highlights
Second harmonic generation (SHG) microscopy represents a very powerful tool for tissue characterization
Special emphasis is placed on identifying which of the methods developed in this purpose over the past years by the digital image processing community are best aligned to human expert opinion in the case of Polarization-resolved SHG (PSHG) micrographs collected under various polarization states of the incident light
For every sample here used 24 PSHG images were involved in the study, 12 corresponding to the considered Linear Polarization States (LPS), and 12 corresponding to the considered Elliptical Polarization States (EPS); more information is provided in the Methods section
Summary
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. 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. Special emphasis is placed on identifying which of the methods developed in this purpose over the past years by the digital image processing community are best aligned to human expert opinion in the case of PSHG micrographs collected under various polarization states of the incident light. SHG (and PSHG) imaging is possible in both forward and backward configurations[20,21], for similar reasons a backscattered PSHG geometry was considered, as presented in the Methods section
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