Tailored 3D microphantoms: An essential tool for quantitative phase tomography analysis of organoids

Abstract

We present a novel approach for benchmarking and validating quantitative phase tomography (QPT) systems using three-dimensional microphantoms. These microphantoms, crafted from biological and imaging data, replicate the optical and structural properties of multicellular biological samples. Their fabrication featuring refractive index modulation at sub-micrometer details is enabled by two-photon polymerization. We showcase the effectiveness of our technique via a round-robin test of healthy and tumoral liver organoid phantoms across three different QPT systems. This test reveals sample- and system-dependent errors in measuring dry mass and morphology. This approach constitutes a development of super phantoms for QPT — test objects that exist in both digital and physical form, replicate both the morphology and relevant aspects of physiology in specimens under healthy or diseased conditions, and underpin the assessment and refinement of imaging technologies and methodologies prior to clinical application.