Comprehensive Characterization of an 'Off/On' Rhodol‐Based Lysosomal Tracker for Orthogonal Cellular Analysis by Confocal Imaging

Autor: Carolina Chieffo, Emiliano Altamura, Layth Ben Trad, Guillaume Pilet, Ofelia Maniti, Thierry Granjon, Saida Mebarek, Peter Strazewski, Michele Fiore
Rok vydání: 2022
Předmět:
Zdroj: ChemBioChem. 24
ISSN: 1439-7633
1439-4227
DOI: 10.1002/cbic.202200513
Popis: Two florescent xanthene-cyanamide lysosomal trackers emitting strongly at ∼525 nm were prepared from fluorescein and rhodol methyl esters in microwave-assisted reactions. Both forms named "off" (nonfluorescent lactam) and "on" (strongly fluorescent ring-opened amide) have been comprehensively characterized out by using a combination of NMR spectroscopy, X-ray analysis, fluorimetry and confocal microscopy. Known rhodamines bearing electron-withdrawing groups (EWGs) exhibit an equilibrium between non-fluorescent (off) and fluorescent (on) depending on the dielectric constant of the medium. Here, cyanamide was introduced as EWG amine into the fluorescein and rhodol framework. Unlike rhodamine-type dyes, the ring-opened forms of fluorescein- and rhodol-cyanamides are stable in protic solvents under circumneutral and basic pH conditions. The osteoblastic cell line MC3T3-E1 from C57BL/6 mouse calvaria was used for confocal imaging where the different organelles and nuclei were distinguished by using an orthogonal combination of fluorescent dyes.
Databáze: OpenAIRE
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