Amber particles as living plant cell markers in flow cytometry

The unique biological properties of amber are well known. Amber particles that penetrate into the cellular matrix can potentially be used as markers of plant cell biological activity by identification of living cells in flow cytometry. However, there have been no studies on effect of amber particles on plant cells. The aim of this study was to determine effect of amber nano- and micro- (5 nm^-3 μm) particles on somatic and gametic cells and to assess the possibility to use amber particles as alive plant cells markers. To reach the aim, fluorescence of cells was determined in the presence of amber particles, and amber components - spirit of amber and sodium succinate dibasic hexahydrate. Cell fluorescence was measured using a BD FACSJazz® cell sorter for several plants species (Argyranthemum frutescens, Cyclamen persicum, Hordeum vulgare and Linum usitatissimum) with and without treatment of amber. Differences between a control cell group (without amber treatment) and treated cell group with amber particles depended on plant species. The presence of amber components (alcohol soluble amber fraction and sodium succinate) in cultivation media mostly had no influence on cell fluorescence. The study showed that amber particles (size 5 nm^-3 μm) can be used as living plant cell markers, as the presence of amber particles in plant cell cultivation media resulted in substantially increased plant cell fluorescence in all investigated species, and there was no detrimental effect of amber particles on plant cells.