Design and characterization of an optical phantom for mesoscopic multimodal fluorescence lifetime imaging and optical coherence elastography.

We developed a novel methodology for manufacturing multimodal, tissue-mimicking phantoms that exhibit both molecular and biomechanical contrast. This methodology leverages the immiscibility of silicone and hydrogels to create solid mesoscale phantoms with localized regions of precisely controlled fluorescence, including fluorescence lifetime properties, and adjustable stiffness, without requiring physical barriers. Mechanical, fluorescent, and optical characterization confirmed the tunability of the phantoms across a range of values relevant to biomedical applications. A macroscale 3D phantom was fabricated, and its properties were validated through fluorescence lifetime imaging (FLI) and optical coherence elastography (OCE). Validation demonstrated the successful tuning of both mechanical and fluorescence lifetime contrasts within a 3D structure, highlighting the feasibility of multimodal FLI-OCE. This new phantom manufacturing process is expected to support the development and validation of new multimodal imaging approaches to study molecular and biomechanical properties of the tumor microenvironment (TME), as well as their impact on therapeutic efficacy, and to enhance targeted therapies.