Lucifer Yellow Uptake in Cells and Protoplasts of Daucas carota Visualized by Laser Scanning Microscopy

The uptake of lucifer yellow CH by suspension-cultured carrot cells and protoplasts has been studied by laser scanning microscopy. This fluorochrome, which does not diffuse across membranes, gradually accumulates in the cell vacuole over a period of hours. In contrast, the central vacuole of protoplasts did not show lucifer yellow fluorescence. The latter was restricted, in protoplasts, to punctate sources in the peripheral cytoplasm. Confocal optics allowed the complexity of the vacuolar system to be dramatically depicted with the laser scanning microscope. Control experiments support the contention that lucifer yellow uptake, as in other eukaryotic systems, occurs via endocytosis.