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Cryo-fixation and associated developments in transmission electron microscopy : a cool future for nematology

Wim Bert (UGent) , Dieter Slos (UGent) , Olivier Leroux (UGent) and Myriam Claeys (UGent)
(2016) NEMATOLOGY. 18(1). p.1-14
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Abstract
At present, the importance of sample preparation equipment for electron microscopy represents the driving force behind major breakthroughs in microscopy and cell biology. In this paper we present an introduction to the most commonly used cryo-fixation techniques, with special attention paid towards high-pressure freezing followed by freeze substitution. Techniques associated with cryofixation, such as immunolocalisation, cryo-sectioning, and correlative light and electron microscopy, are also highlighted. For studies that do not require high resolution, high quality results, or the immediate arrest of certain processes, conventional methods will provide answers to many questions. For some applications, such as immunocytochemistry, three-dimensional reconstruction of serial sections or electron tomography, improved preservation of the ultrastructure is required. This review of nematode cryo-fixation highlights that cryo-fixation not only results in a superior preservation of fine structural details, but also underlines the fact that some observations based on results solely obtained through conventional fixation approaches were either incorrect, or otherwise had severe limitations. Although the use of cryo-fixation has hitherto been largely restricted to model organisms, the advantages of cryo-fixation are sufficiently self-evident that we must conclude that the cryo-fixation method is highly likely to become the standard for nematode fixation in the near future.
Keywords
freeze substitution, high pressure freezing, electron microscopy, immuno-electron microscopy, specimen preparation, tissue sections, ultrastructural preservation, APHELENCHUS-AVENAE NEMATODA, ACROBELES-COMPLEXUS NEMATODA, MICROBIAL FEEDING NEMATODE, CAENORHABDITIS-ELEGANS, FREEZE-SUBSTITUTION, C-ELEGANS, 3-DIMENSIONAL RECONSTRUCTION, CORRELATIVE LIGHT, PRISTIONCHUS-PACIFICUS, ULTRATHIN CRYOSECTIONS

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Citation

Please use this url to cite or link to this publication:

MLA
Bert, Wim et al. “Cryo-fixation and Associated Developments in Transmission Electron Microscopy : a Cool Future for Nematology.” NEMATOLOGY 18.1 (2016): 1–14. Print.
APA
Bert, W., Slos, D., Leroux, O., & Claeys, M. (2016). Cryo-fixation and associated developments in transmission electron microscopy : a cool future for nematology. NEMATOLOGY, 18(1), 1–14.
Chicago author-date
Bert, Wim, Dieter Slos, Olivier Leroux, and Myriam Claeys. 2016. “Cryo-fixation and Associated Developments in Transmission Electron Microscopy : a Cool Future for Nematology.” Nematology 18 (1): 1–14.
Chicago author-date (all authors)
Bert, Wim, Dieter Slos, Olivier Leroux, and Myriam Claeys. 2016. “Cryo-fixation and Associated Developments in Transmission Electron Microscopy : a Cool Future for Nematology.” Nematology 18 (1): 1–14.
Vancouver
1.
Bert W, Slos D, Leroux O, Claeys M. Cryo-fixation and associated developments in transmission electron microscopy : a cool future for nematology. NEMATOLOGY. 2016;18(1):1–14.
IEEE
[1]
W. Bert, D. Slos, O. Leroux, and M. Claeys, “Cryo-fixation and associated developments in transmission electron microscopy : a cool future for nematology,” NEMATOLOGY, vol. 18, no. 1, pp. 1–14, 2016.
@article{8087233,
  abstract     = {At present, the importance of sample preparation equipment for electron microscopy represents the driving force behind major breakthroughs in microscopy and cell biology. In this paper we present an introduction to the most commonly used cryo-fixation techniques, with special attention paid towards high-pressure freezing followed by freeze substitution. Techniques associated with cryofixation, such as immunolocalisation, cryo-sectioning, and correlative light and electron microscopy, are also highlighted. For studies that do not require high resolution, high quality results, or the immediate arrest of certain processes, conventional methods will provide answers to many questions. For some applications, such as immunocytochemistry, three-dimensional reconstruction of serial sections or electron tomography, improved preservation of the ultrastructure is required. This review of nematode cryo-fixation highlights that cryo-fixation not only results in a superior preservation of fine structural details, but also underlines the fact that some observations based on results solely obtained through conventional fixation approaches were either incorrect, or otherwise had severe limitations. Although the use of cryo-fixation has hitherto been largely restricted to model organisms, the advantages of cryo-fixation are sufficiently self-evident that we must conclude that the cryo-fixation method is highly likely to become the standard for nematode fixation in the near future.},
  author       = {Bert, Wim and Slos, Dieter and Leroux, Olivier and Claeys, Myriam},
  issn         = {1388-5545},
  journal      = {NEMATOLOGY},
  keywords     = {freeze substitution,high pressure freezing,electron microscopy,immuno-electron microscopy,specimen preparation,tissue sections,ultrastructural preservation,APHELENCHUS-AVENAE NEMATODA,ACROBELES-COMPLEXUS NEMATODA,MICROBIAL FEEDING NEMATODE,CAENORHABDITIS-ELEGANS,FREEZE-SUBSTITUTION,C-ELEGANS,3-DIMENSIONAL RECONSTRUCTION,CORRELATIVE LIGHT,PRISTIONCHUS-PACIFICUS,ULTRATHIN CRYOSECTIONS},
  language     = {eng},
  number       = {1},
  pages        = {1--14},
  title        = {Cryo-fixation and associated developments in transmission electron microscopy : a cool future for nematology},
  url          = {http://dx.doi.org/10.1163/15685411-00002943},
  volume       = {18},
  year         = {2016},
}

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