Blood flow imaging is widely applied in photodynamic therapy (PDT) to provide vascular morphological and statistical parameters. This approach relies on the intensity of time-domain signal differences between blood vessels and background tissues; therefore, it often ignores differences within the vasculature and cannot accommodate abundant structural information. This study proposes a multi-level optical angiography (MOA) method for PDT. It can enhance capillaries and image vessels at different levels by measuring the signal frequency shift associated with red blood cell motion. The experimental results regarding the PDT-induced chorioallantoic membrane model showed that the proposed method could not only perform multi-level angiography but also provide more accurate quantitative information regarding various vascular parameters. This MOA method has potential applications in PDT studies.
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