Blue-conversion of organic dyes produces artifacts in multicolor fluorescence imaging
Dohyeon Kim, Yeonho Chang, Soyeon Park, Min Gyu Jeong, Yonghoon Kwon, Kai Zhou, Jungeun Noh, Yunkyu Choi, Triet Minh Hong, Young‐Tae Chang, Sung Ho Ryu
Abstract
Multicolor fluorescence imaging is a powerful tool visualizing the spatiotemporal relationship among biomolecules. Here, we report that commonly employed organic dyes exhibit a blue-conversion phenomenon, which can produce severe multicolor image artifacts leading to false-positive colocalization by invading predefined spectral windows, as demonstrated in the case study using EGFR and Tensin2. These multicolor image artifacts become much critical in localization-based superresolution microscopy as the blue-converted dyes are photoactivatable. We provide a practical guideline for the use of organic dyes for multicolor imaging to prevent artifacts derived by blue-conversion.