Immunoelectron Microscopy of Fission Yeast Using High Pressure Freezing

M. Onomi; N. Kamasawa; T. Takagi; M. Osumi

doi:10.5685/plmorphol.12.20

M. Onomi, N. Kamasawa, T. Takagi, M. Osumi

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Keywords: S. pombe, High Pressure Freezing, Immunoelectron Microscopy, Ultrastructure

JOURNAL FREE ACCESS

2000 Volume 12 Issue 1 Pages 20-31

DOI https://doi.org/10.5685/plmorphol.12.20

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Published: 2000 Received: - Available on J-STAGE: June 28, 2010 Accepted: - Advance online publication: - Revised: -
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Date of correction: June 28, 2010 Reason for correction: - Correction: PDF FILE Details: -

Date of correction: June 28, 2010 Reason for correction: - Correction: KEYWORD Details: Wrong : Key Words: S. pombe, High Pressure Freezing, Immunoelectron Microscopy, Ultrastructure
Right : S. pombe, High Pressure Freezing, Immunoelectron Microscopy, Ultrastructure

Date of correction: June 28, 2010 Reason for correction: - Correction: CITATION Details: Wrong : E
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Right : Baba, M., Osumi, M.(1987)Transmission and scanning electron microscopic examination of intracellular organelles in freeze-substituted Kloeckera and Saccharomyces cerevisiae cells. J. Electron Microsc. Tech. 5: 246-261
Ding, B., Turgeon, R., Parthasarathy, MV.(1991)Efect of high-pressure freezing on plant microfilament bundles. J. Microsc. 165: 367-376
Ding, R., McDonald, K.L., McIntosh, J.R.(1993)Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. J. Cell Biol. 120: 141-151
Driouich, A., Zhang, GF., Staehelin, LA.(1993)Effect of brefeldin A on the structure of the golgi apparatus and on the synthesis and secretion of proteins and polysaccharides in sycamore maple(Acer pseudoplatanus)suspension-cultured cells. Plant Physiol. 101: 1363-1373
Erk, I., Nicolas, G., Caroff, A., Lepault, J.(1998)Electron microscopy of frozen biological objects: a study using cryosectioning and cryosubstitution. J. Microsc. 189: 236-248
Fujimoto, K.(1995)Freeze-fracture replica electron microscopy combined with SDS digestion for cytochemical labeling of integral membrane proteins. J. Cell Sci. 108: 3443-3449
Hohenberg, H., Mannweiler, K., Meuller, M.(1994)High-pressure freezing of cell suspensions in cellulose capillary tubes. J. Microsc. 175: 34-43
Humbel, B.M., Konomi, M., Takagi, T., Kamasawa, N., Ishijima, S.A., Osumi, M.(2001)In situ localisation of β -glucans in the cell wall of Schizosaccharomyces pombe. Yeast 18: 1-13
Kamasawa, N., Yoshida, T., Ueda, M., Tanaka, A., Osumi, M.(1999)Three dimensional analysis of protein aggregate body in Saccharomyces cerevisiae. J. Electron Microsc. 48: 173-176
Kanbe, T., Akashi, T., Tanaka, K.(1994)Changes in the distribution of F-actin in the fission yeast Schizosaccharomyces pombe by arresting growth in distilled water: correlative studies with fluorecence and electron microscopy. J. Electron Microsc. 43: 20-24
Kaneko, Y., Walther, P.(1995)Comparison of ultrastructure of germinating pea leaves prepared by high-pressure freezing-freeze substitution and conventional chemical fixation. J. Electron Microsc. 44: 104-109
McDonald, K., Morphew, M. K.(1993)Improved preservation of ultrastructure in difficult-to-fix organisms by high pressure freezing and freeze substitution: I. Dorsophila melanogaster and Strongylocentrotus purpuratus embryos. Microsc. Res. Tech. 24: 465-473
Michel, M., Hillmann, T., Meuller, M.(1991)Cryosectioning of plant material frozen at high pressure. J. Microsc. 163: 3-18
Moor, H., Riehle, U.(1968)Snap freezing under high pressure: a new fixation technique for freeze-etching. Proc. 4th Eur. Reg. Conf. Electron Microsc., ed. D. S. Bocciarelli, Rome, 2: 445-446
Moor, H.(1987)Theory and practice of high pressure freezing. In Cryotechniques in Biological Electron microscopy, eds. R.A. Steinbrecht, K. Zierold(Springer-Verlag, Berlin heidelberg), pp. 175-191
Monaghan, P., Perusinghe, N., Meuller, M.(1998)High-pressure freezing for immunocytochemistry. J. Microsc. 192: 248-258
Naito, N., Yamada, N., Kobori, H., Osumi, M.(1991)Contrast enhancement by ruthenium tetroxide for observation of the ultrastructure of yeast cells. J. Electron Microsc. 40: 416-419
Osumi, M., Baba, M., Naito, N., Taki, A., Yamada, N., Nagatani, T.(1988)High resolution, low voltage scanning electron microscopy of uncoated yeast cells fixed by the freeze-substitution method. J. Electron Microsc. 37: 17-30
Osumi, M.(1998)The ultrastructure of yeast: cell wall structure and formation. Micron 29: 207-233
Osumi, M., Konomi, M., Takagi, T.(2000)High pressure freezing is a powerful technique for immunoelectron microscopy and low temperature SEM in yeasts. Proceedings of the10th International Symposium on Yeasts(Papendal, Arnhem, The Netherlands), pp. 352-353
Robinow, C.F., Hyams, J.S.(1989)General cytology of fission yeasts. In Molecular Biology of the Fission Yeast, eds. A. Nasim, P. Young, B. F. Johnson(Academic Press Inc.), pp. 273-330
Sawaguchi, A., Ide, S., Kawano, J., Nagaike, R., Oinuma, T., Tojo, H., Okamoto, M., Suganuma, T.(1999)Reappraisal of potassium permanganate oxidation applied to Lowicryl K4M embedded tissues processed by high pressure freesing/freeze substitution, with special reference to differential staining of the zymogen granules of rat gastric chief cells. Arch. Histol. Cytol. 62: 447-458
Shibasaki, Y., Kamasawa, N., Shibasaki, S., Zou, W., Murai, T., Ueda, M., Tanaka, A., Osumi, M.(2000)Cytochemical evalution of localization and secretion of heterologous enzyme displayed on yeast cell surface. FEMS Microbiol. Lett. 192: 243-248
Steinbrecht, R.A. Meuller, M.(1987)Freeze-substitution and freeze-drying. In Cryotechniques in Biological Electron microscopy, eds. R.A. Steinbrecht, K. Zierold(Springer-Verlag, Berlin heidelberg), pp. 149-172
Studer, D., Michel, M., Meuller, M.(1989)High pressure freezing comes of age. Scanning Microsc. 3: 253-269
Tiedemann, J. Hohenberg, H., Kollmann, R.(1998)High-pressure freezing of plant cells cultured in cellulose microcapillaries. J. Microsc. 189: 163-171
Wild, P., Schraner, E.M., Adler, H., Humbel, B.M .(2000)Enhanced resolution of membranes in cultured cells by cryoimmobilization and freeze-substitution. Microsc. Res. Tech.(in press)

Abstract

Summary: A high pressure freeze-substitution method was developed for immunoelectron microscopy(immuno-EM)of S. pombe. The method used a low concentration of OsO4 or uranyl acetate(UA)provided a high contrast in the specimen, while retaining the fine structure of the cells. In the image fixed with 0.01% OsO4-acetone, the cell wall and cytoskeleton were visible, while 0.1% UA-acetone was superior in the membranous structure and cytoplasm. The new method makes it possible to visualize the ultrastructure and localization of the material on one thin section for the immuno-EM.

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