Biopolym. Cell. 2021; 37(6):447-458.
http://dx.doi.org/10.7124/bc.000A6A
Molecular and Cell Biotechnologies
Benzoxazole styrylcyanine dye as a fluorescent probe for functional amyloid visualization in Staphylococcus aureus ATCC25923 biofilm
1Chernii S. V., 1Moshynets O. V., 1Aristova D. I., 1Kryvorotenko D. V., 1Losytskyy M. Yu., 1, 2Iungin O. S., 1Yarmoluk S. M., 1Volynets G. P.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03143
  2. Kyiv National University of Technologies and Design
    2, Nemirovich-Danchenko Str., Kyiv, Ukraine, 01011

Abstract

Aim. Synthesis and characterization of styrylcyanine dye as a potential fluorescent probe for the detection in vitro of pathological amyloid fibrils and functional amyloid in S. aureus biofilm. Methods. Chemical synthesis, fluorescence spectroscopy, irradiation with a visible light source, confocal laser scanning microscopy, fluorescence microscopy. Results. Styrylcyanine dye is low fluorescent when free, but in the presence of amyloid fibrils in vitro shows an increase in the emission intensity up to 214 times depending on the amyloidogenic protein structure; the most pronounced enhancement was observed for fibrils of beta-lactoglobulin. Photostability of the dye in the free state is low, but binding to amyloid fibrils results in a strong increase of dye photostability. Upon staining S. aureus biofilm, the dye stains extracellular components of the biofilm matrix and does not penetrate the cell. Conclusion. This dye is suggested to visualize the functional amyloids of S. aureus biofilm with a red emission.
Keywords: fluorescence microscopy, styryl dyes, S. aureus, functional amyloids, bacterial biofilm, laser scanning confocal microscopy
Full text: (PDF, in English)

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