Fluorescence imaging for surgical guidance is a proven modality that allows for visualization of fluorescent markers in numerous biological imaging applications. As the field continues to develop there is an urgent need for fluorescence-imaging standards that enable system characterization and performance monitoring. Here, we present a long-term stable standard that mimics indocynine green (ICG) spectral behavior and is capable of probing the sensitivity of commercially available imaging systems. The standard is composed of three different tests: a varying concentration sensitivity test (0.1nM-3000nM), a tissue-equivalent-depth sensitivity test (0.2mm-7mm), and a 1951 USFA fluorescence resolution test. The fluorescence standard platform developed can incorporate 3-D printing with tissue-equivalent layers, enabling a wide variety of anatomical-specific designs. The manufacturing capabilities developed allow for incorporation of different fluorophores for applications beyond ICG and NIR.
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