Abstract
Fluorescence microscopy is the essential technique for investigation of the intracellular distribution of macromolecules and various organelles also in yeast cells. In this chapter, detailed practical procedures for fluorescence microscopic observations developed or adopted in our laboratory are described. These include labeling of the cell wall and chitin, F-actin structures, nuclear and mitochondrial DNA, and two different procedures for investigation of yeast cells by immunofluorescence. In addition, our experience with multicolor labeling experiments is introduced and discussed.
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Kilmartin, J. V. and Adams, A. E. M. (1984) Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J. Cell Biol. 98, 922–933.
Pringle, J. R., Adams, A. E. M., Drubin, D. G., and Haarer, B. K. (1991) Immunofluorescence methods for yeast. Methods Enzymol. 194, 565–602.
Hasek, J. and Streiblová, E. (1996) Fluorescence microscopy methods, in Methods in Molecular Biology (Evans, I. H. and Walker, J. M., eds.), Humana Press, Totowa, NJ, pp. 11–15.
Kohlwein, S. D. (2000) The beauty of the yeast: live cell microscopy at the limits of optical resolution. Microscopy Res. Tech. 51, 511–529.
Burd, C. G. (2000) Visualizing protein dynamics in yeast with green fluorescent protein. Methods Enzymol. 327, 61–69.
Fritze, C. E. and Anderson, T. R. (2000) Epitope tagging: general method for tracking recombinant proteins. Methods Enzymol. 327, 3–16.
Prein, B., Natter, K., and Kohlwein, S. D. (2000) A novel strategy for constructing N-terminal chromosomal fusions to green fluorescent protein in the yeast Saccharomyces cerevisiae. FEBS Lett. 485, 29–34.
Vida, T. A. and Emr, S. D. (1995) A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J. Cell Biol. 128, 779–792.
Koning, A. J., Lum, P. Y., Williams, J. M., and Wright, R. (1993) DiOC6 staining reveals organelle structure and dynamics in living yeast cells. Cell Motil Cytoskeleton 25, 111–128.
Bereiter-Hahn, J., Seipel, K. H., Voth, M., and Ploem, J. S. (1983) Fluorimetry of mitochondria in cells vitally stained with DASPMI or rhodamine 6 GO. Cell Biochem Funct. 1, 147–155.
Streiblová E. (1984) The yeast cell wall: a marker system for cell cycle controls, in The Microbial Cell Cycle (Nurse, P. and Streiblová., E. eds.), CRC Press, Boca Raton, FL.
Wieland, T. (1986) Peptides in Poisonous Amanita Mushrooms. Springer Verlag, Heidelberg, FRG.
Jirincová, H., Vavricková, P., Palecek, J., and Hasek, J. (1998) A new monoclonal antibody against Rpg1p. Folia Biol. 44, 73.
Valá, L., Trachsel, H., Hasek, J., and Ruis, H. (1998) Rpg1p, the Saccharomyces cetrevisiae homologue of the largest subunit of mammalian translation initiation factor 3, is required for translational activity. J. Biol. Chem. 273, 21253–21260.
Valá, L., Hasek, J., Nielsen, K. H., and Hinnebusch, A.G. (2001) Dual function of eIF3j/Hcr1p in processing 20S Pre-rRNA and translation initiation. J. Biol. Chem. 276, 43351–43360.
Nagata, Y. and Burger, M. M. (1974) Wheat germ agglutinin. Molecular characteristics and specificity for sugar binding. J. Biol. Chem. 249, 3116–3122.
Kohlwein, S. D., Eder, S., Oh, C. S., Martin, C. E., Gable, K., Bacikova, D., and Dunn, T. (2001) Tsc13p is required for fatty acid elongation and localizes to a novel structure at the nuclear-vacuolar interface in Saccharomyces cerevisiae. Mol Cell Biol. 21, 109–125.
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© 2006 Humana Press Inc., Totowa, NJ
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Hašek, J. (2006). Yeast Fluorescence Microscopy. In: Xiao, W. (eds) Yeast Protocol. Methods in Molecular Biology, vol 313. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-958-3:085
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DOI: https://doi.org/10.1385/1-59259-958-3:085
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-437-1
Online ISBN: 978-1-59259-958-5
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