Application of Time-Resolved Fluorescence Microscopy for Enhancing the Selectivity of Fluorogenic Dyes of the Arylidene–Imidazolone Series toward the Endoplasmic Reticulum

Objective: A number of previously synthesized fluorogenic arylidene-imidazolones, which predominantly stain the endoplasmic reticulum (ER) of living cells, were studied by time-resolved fluorescence microscopy. It was suggested that the use of fluorescence microscopy of this type can enhance the sel...

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Bibliographic Details
Published in:Russian journal of bioorganic chemistry 2024-10, Vol.50 (5), p.1896-1903
Main Authors: Gilvanov, A. R., Smirnov, A. Yu, Krasnova, S. A., Solovyev, I. D., Savitsky, A. P., Bogdanova, Yu. A., Baranov, M. S.
Format: Article
Language:English
Subjects:
Biochemistry
Biomedical and Life Sciences
Biomedicine
Bioorganic Chemistry
Decay
Dyes
Endoplasmic reticulum
Fluorescence
Heterocyclic compounds
Life Sciences
Microscopy
Organelles
Organic Chemistry
Staining
Time correlation functions
Time measurement
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Summary:Objective: A number of previously synthesized fluorogenic arylidene-imidazolones, which predominantly stain the endoplasmic reticulum (ER) of living cells, were studied by time-resolved fluorescence microscopy. It was suggested that the use of fluorescence microscopy of this type can enhance the selectivity of ER staining. Methods: The lifetimes of arylidene-imidazolone compounds in set of solvents with different polarity were measured with time-correlated single photon counting spectroscopy. Live HeLa Kyoto cells were stained with studied dyes and analyzed with time-resolved fluorescence microscopy. Results and discussion: It was found that most of the studied compounds show bi- or triexponential fluorescence decay patterns in solutions. In live cell culture dye ( I ) showed a monoexponential decay pattern, while dyes ( II–IV ) were better fitted by a biexponential function. Dyes ( I ), ( III ), and ( IV ) stained both ER and adiposomes, while dye ( II ) stained only ER. Conclusions: It is shown that under FLIM conditions discriminative filtering of cellular organelles stained with studied fluorogenic dyes is possible and applicable if the difference of mean amplitude-weighted lifetime is more than 0.1 ns, thus increasing the selectivity of ER staining in live cells.
ISSN:1068-1620
1608-330X
DOI:10.1134/S1068162024050315