A series of optimized protocols to isolate vacuoles from both yeast and plant cells, and to characterize the purified organelles at a functional and structural level, are described. For this purpose, we took advantage of the combined use of cell fractionation techniques with different fluorescence-based approaches namely flow cytometry, fluorescence microscopy and spectrofluorimetry. These protocols altogether constitute valuable tools for the study of vacuole structure and function, as well as for the high-throughput screening of drug libraries to identify new molecules that target the vacuole.
CITATION STYLE
Rodrigues, J. M. P., Pereira, C. S., Fontes, N., Gerós, H., & Côrte-Real, M. (2018). Flow cytometry and fluorescence microscopy as tools for structural and functional analysis of vacuoles isolated from yeast and plant cells. In Methods in Molecular Biology (Vol. 1789, pp. 101–115). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7856-4_8
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