Blue-conversion of organic dyes produces artifacts in multicolor fluorescence imaging

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...

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Bibliographic Details
Published in:Chemical science (Cambridge) 2021-06, Vol.12 (25), p.866-8667
Main Authors: Kim, Do-Hyeon, Chang, Yeonho, Park, Soyeon, Jeong, Min Gyu, Kwon, Yonghoon, Zhou, Kai, Noh, Jungeun, Choi, Yun-Kyu, Hong, Triet Minh, Chang, Young-Tae, Ryu, Sung Ho
Format: Article
Language:English
Subjects:
Biomolecules
Chemistry
Conversion
Dyes
Fluorescence
Imaging
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Summary: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. Blue-conversion, a photooxidative conversion leading to the hypsochromic shift of absorption and emission spectra, occurs in popular organic dyes under conventional laser illumination and produces severe artifacts in multicolor fluorescence imaging.
ISSN:2041-6520
2041-6539
DOI:10.1039/d1sc00612f