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Blue-conversion of organic dyes produces artifacts in multicolor fluorescence imaging
- Source :
- Chemical Science
- Publication Year :
- 2021
-
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.<br />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.
- Subjects :
- 0301 basic medicine
Fluorescence-lifetime imaging microscopy
Materials science
Colocalization
Nanotechnology
General Chemistry
Superresolution
Practical guideline
03 medical and health sciences
Chemistry
030104 developmental biology
0302 clinical medicine
Microscopy
embryonic structures
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 20416520
- Volume :
- 12
- Issue :
- 25
- Database :
- OpenAIRE
- Journal :
- Chemical science
- Accession number :
- edsair.doi.dedup.....cece5739308eb4306f56f6b3016386b3