This method tracks structures in yeast cells in three dimensions over many minutes, allowing us to study the dynamics of intracellular organelles and compartments. 4D imaging ensures that we can draw reliable conclusions about intracellular dynami
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This protocol describes the analysis of fluorescently labeled intracellular compartments in budding yeast using multi-color 4D (time-lapse 3D) confocal microscopy. The imaging parameters are chosen to capture adequate signals while limiting photodamage. Custom ImageJ plugins allow labeled structures to be tracked and quantitatively analyzed.