Why the study?
Label-free cardiac second harmonic generation imaging struggles to specifically identify collagen versus myosin because both are harmonophores generating signals.
PSR staining followed by multiphoton excitation fluorescence imaging provides a robust and high signal-to-noise method for quantifying cardiac fibrosis in preclinical models.
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Provides reliable fibrosis quantification in preclinical models; leaves open clinical translation pending validation.
Traditional methodologies for fibrosis quantification involve histological measurements, staining with Masson's trichrome and picrosirius red (PSR), and label-free imaging using second harmonic generation (SHG). The difficulty of label-free cardiac SHG imaging is that both collagen (i.e., collagen 1 fibrils) and myosin are harmonophores that generate SHG signals, and specific identification of either collagen or myosin is difficult to achieve. Here we present an alternate method of quantifying cardiac fibrosis by using PSR staining followed by multiphoton excitation fluorescence imaging. Our data from the deoxycorticosterone model of cardiac fibrosis shows that this imaging method and downstream analyses, including background correction, are robust and easy to perform. These advantages are due to the high signal-to-noise ratio provided by PSR in areas of collagen fibers. Furthermore, the hyperspectral and fluorescence lifetime information of PSR-stained area of fibrosis shows better quantification can eventually be obtained using more complex instrumentation.
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Jones et al. (2023) studied this question.
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