Novel fluorescent dyes based on oligopropylamines for the in vivo staining of eukaryotic unicellular algae |
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Authors: | VV Annenkov EN Danilovtseva SN Zelinskiy TN Basharina TA Safonova ES Korneva YV Likhoshway MA Grachev |
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Institution: | Limnological Institute of Siberian Branch, Russian Academy of Sciences, 660033 Irkutsk, Russia |
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Abstract: | Weakly basic fluorescent dyes are used to visualize organelles within live cells due to their affinity to acidic subcellular organelles. In particular, they are used to stain the silica deposited in the silica deposition vesicles (SDVs) of diatoms during the course of their frustule synthesis. This study involved the synthesis of fluorescent dyes derived from oligopropylamines, compounds similar to those found in diatoms. The dyes were obtained by reacting oligopropylamines with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole. The reaction was realized using methylated oligopropylamines with two or three nitrogen atoms and yielded two novel fluorescent dyes: NBD-N2 and NBD-N3. The dyes appeared to be highly efficient in the in vivo staining of growing siliceous frustules of diatoms at concentrations at least 10 times lower than those required for staining with HCK-123. NBD-N3 also efficiently stained other subcellular vesicles of eukaryotic unicellular algae. NBD-N2 stained only growing diatom frustules, whereas NBD-N3 also stained various subcellular organelles of different eukaryotic unicellular algae. NBD-N2 and NBD-N3 were not removed from stained diatom frustules by drastic treatments with H2SO4 and H2O2. Fluorescent silica can also be obtained by its chemical precipitation in the presence of NBD-N2 and NBD-N3. |
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Keywords: | Fluorescent dye Oligopropylamines Eukaryote Diatom Silica deposition vesicle |
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