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We demonstrated the ability of fiber-based Fluorescence Lifetime Imaging (FLIm) guided Raman spectroscopy to monitor the quality of engineered vascular grafts with high speed and specificity. We report FLIm guided Raman imaging as an effective multimodal technique to evaluate scaffold cross-linking and localized calcification. Current results indicate that the lifetime of AR-BP shortens upon GA cross-linking,and Raman spectroscopy reveals secondary structural changes occurring in the Amide I region of cross-linked pericardia. GA fixed vascular grafts are prone to calcification, an effect linked to graft failure. The calcified regions exhibited shorter lifetimes in fluorescence spectral bands ranging from 380 to 455 nm and Raman spectra exhibited the specific hydroxyapatite signature at 960 cm-1 co-localized with these lower lifetime regions. We conclude that FLIm guided Raman imaging can detect cross-linking signatures and areas of calcification in tissue with biochemical specificity.
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Tanveer Ahmed Shaik, Alba Alfonso Garcia, Xiangnan Zhou, Anne K. Haudenschild, Christoph Krafft, Laura Marcu, Jürgen Popp, "FLIm guided Raman imaging for detecting bovine pericardium cross-links and calcification (Conference Presentation)," Proc. SPIE 11215, Diagnostic and Therapeutic Applications of Light in Cardiology 2020, 112150I (9 March 2020); https://doi.org/10.1117/12.2545632