Moshynets, Olena and Chernii, Svitlana and Chernii, Viktor and Losytskyy, Mykhaylo and Karakhim, Serhii and Czerwieniec, Rafal and Pekhnyo, Vasyl and Yarmoluk, Sergiy and Kovalska, Vladyslava (2020) Fluorescent beta-ketoenole AmyGreen dye for visualization of amyloid components of bacterial biofilms. METHODS AND APPLICATIONS IN FLUORESCENCE, 8 (3): 035006. ISSN 2050-6120,
Full text not available from this repository. (Request a copy)Abstract
Green-emitting water-soluble amino-ketoenole dye AmyGreen is proposed as an efficient fluorescent stain for visualization of bacterial amyloids in biofilms and the detection of pathological amyloids in vitro. This dye is almost non-fluorescent in solution, displays strong green emission in the presence of amyloid fibril of proteins. AmyGreen is also weakly fluorescent in presence to biomolecules that are components of cells, extracellular matrix or medium: nucleic acids, polysaccharides, lipids, and proteins. Thus, the luminescence turn-on behavior of AmyGreen can be utilized for visualization of amyloid components of bacterial biofilm extracellular matrix. Herein we report the application of AmyGreen for fluorescent staining of a number of amyloid-contained bacteria biofilms produced by Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa,Bordetella avium, and Staphylococcus aureus. The effectiveness of AmyGreen was compared to traditional amyloid sensitive dye Thioflavine T. The main advantage of AmyGreen (concentration 10(-5) M) is a higher sensitivity in the visualization of amyloid biofilm components over Thioflavine T (10(-4) M) as it was revealed when staining E. coli and K. pneumoniae bacterial biofilms. Besides, AmyGreen displays lower cross-selectivity to nucleic acids as demonstrated both in in-solution experiments and upon staining of eukaryotic human mesenchymal stem cells used as amyloid-free negative control over amyloid-rich bacterial biofilms. The results point to a lower risk of false-positive response upon determination of amyloid components of bacterial biofilm using AmyGreen. Co-staining of biofilm by AmyGreen and cellulose sensitive dye Calcofluor White show difference in their staining patterns and localization, indicating separation of polysaccharide-rich and amyloid-rich regions of investigated biofilms. Thus, we suggest the new AmyGreen stain for visualization and differentiation of amyloid fibrils in bacterial biofilms to be used solely and in combination with other stains for confocal and fluorescence microscopy analysis.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROTEIN; AGGREGATION; EXPRESSION; CELLULOSE; ADHESINS; fluorescent detection; beta-ketoenole dye; functional amyloids; bacterial biofilm; laser scanning confocal microscopy |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Physikalische und Theoretische Chemie Chemistry and Pharmacy > Institut für Physikalische und Theoretische Chemie > Chair of Chemistry III - Physical Chemistry (Molecular Spectroscopy and Photochemistry) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 19 Mar 2021 09:07 |
| Last Modified: | 19 Mar 2021 09:07 |
| URI: | https://pred.uni-regensburg.de/id/eprint/44297 |
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