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High‐pressure freezing of cell suspensions in cellulose capillary tubes
Abstract
Summary: A procedure for efficient cryoimmobilization of large volumes of cell suspensions or micro‐organisms by high‐pressure freezing is described. This procedure uses transparent, porous cellulose capillary tubes with an inner diameter of 200 μm, into which the suspensions are drawn by capillary action. The tubes are processed by high‐pressure freezing and freeze‐substitution as if they were tissue samples. Centrifugation of suspensions at low temperatures is no longer necessary and cryopreparation is greatly facilitated. A very high yield of adequately frozen specimens is obtained due to the constant, defined sample geometry. This approach can also be used to process suspensions by conventional chemical fixation, eliminating the need to embed pellets in low‐melting‐point agarose, for example, prior to chemical fixation. The preparation procedure is demonstrated with suspensions of nematodes, paramecia and bacteria.
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Citations
Cited by
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Kapteyn, J. C., Hoyer, L. L., Hecht, J. E., Müller, W. H., Andel, A., Verkleij, A. J., Makarow, M., Van Den Ende, H., & Klis, F. M. (2000) · Molecular Microbiology · doi:10.1046/j.1365-2958.2000.01729.x · Cited by 328
The Gram-Positive Bacterial Cell Wall
Rohde, M. (2019) · Microbiology Spectrum · doi:10.1128/microbiolspec.gpp3-0044-2018 · Cited by 319
Cell differentiation and morphogenesis in the colony-forming choanoflagellate Salpingoeca rosetta
Dayel, M. J., Alegado, R. A., Fairclough, S. R., Levin, T. C., Nichols, S. A., McDonald, K., & King, N. (2011) · Developmental Biology · doi:10.1016/j.ydbio.2011.06.003 · Cited by 277
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Leisinger, T., Bader, R., Hermann, R., Schmid-Appert, M., & Vuilleumier, S. (1994) · Biodegradation · doi:10.1007/bf00696462 · Cited by 73
Studer, D., Zhao, S., Chai, X., Jonas, P., Graber, W., Nestel, S., & Frotscher, M. (2014) · Nature Protocols · doi:10.1038/nprot.2014.099 · Cited by 31
Metadata
- Authors
- HOHENBERG, H., MANNWEILER, K., & MÜLLER, M.
- Venue
- Journal of Microscopy
- Publisher
- Wiley
- Published
- 1994-07
- Volume
- 175
- Issue
- 1
- Pages
- 34-43
- ISSN
- 0022-2720 (ISSN-L)
- 1365-2818
- OpenAlex
- W2083895131
- Cited by
- 234
- References
- 0
- FWCI
- 7.37
- Type
- journal article
- Language
- en
- Open access
- closed
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- HOHENBERG, H., MANNWEILER, K., & MÜLLER, M. (1994). High‐pressure freezing of cell suspensions in cellulose capillary tubes. Journal of Microscopy. https://doi.org/10.1111/j.1365-2818.1994.tb04785.x
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
