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Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa

Ellen Lambrecht (UGent), Julie Baré (UGent), Myriam Claeys (UGent), Natascha Chavatte, Wim Bert (UGent), Koen Sabbe (UGent) and Kurt Houf (UGent)
(2015) BIOTECHNIQUES. 58(4). p.181-188
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Abstract
Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en-and excystment dynamics is a major challenge because no detailed protocols for ultrastructural analysis of cysts are currently available. Transmission electron microscopy (TEM) is ideally suited for those analyses; however, conventional TEM protocols are not satisfactory for cysts of free-living protozoa. Here we report on the design and testing of four protocols for TEM sample preparation of cysts. Two protocols, one based on chemical fixation in coated well plates and one on high-pressure freezing, were selected as the most effective for TEM-based ultrastructural studies of cysts. Our protocols will enable improved analysis of cyst structure and a better understanding of bacterial survival mechanisms in cysts.
Keywords
ultrastructural analysis, intracellular survival, protozoan cysts, transmission electron microscopy, ACANTHAMOEBA-CASTELLANII, SURVIVAL, NEMATODA

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Chicago
Lambrecht, Ellen, Julie Baré, Myriam Claeys, Natascha Chavatte, Wim Bert, Koen Sabbe, and Kurt Houf. 2015. “Transmission Electron Microscopy Sample Preparation Protocols for the Ultrastructural Study of Cysts of Free-living Protozoa.” Biotechniques 58 (4): 181–188.
APA
Lambrecht, Ellen, Baré, J., Claeys, M., Chavatte, N., Bert, W., Sabbe, K., & Houf, K. (2015). Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa. BIOTECHNIQUES, 58(4), 181–188.
Vancouver
1.
Lambrecht E, Baré J, Claeys M, Chavatte N, Bert W, Sabbe K, et al. Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa. BIOTECHNIQUES. 2015;58(4):181–8.
MLA
Lambrecht, Ellen, Julie Baré, Myriam Claeys, et al. “Transmission Electron Microscopy Sample Preparation Protocols for the Ultrastructural Study of Cysts of Free-living Protozoa.” BIOTECHNIQUES 58.4 (2015): 181–188. Print.
@article{5938620,
  abstract     = {Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en-and excystment dynamics is a major challenge because no detailed protocols for ultrastructural analysis of cysts are currently available. Transmission electron microscopy (TEM) is ideally suited for those analyses; however, conventional TEM protocols are not satisfactory for cysts of free-living protozoa. Here we report on the design and testing of four protocols for TEM sample preparation of cysts. Two protocols, one based on chemical fixation in coated well plates and one on high-pressure freezing, were selected as the most effective for TEM-based ultrastructural studies of cysts. Our protocols will enable improved analysis of cyst structure and a better understanding of bacterial survival mechanisms in cysts.},
  author       = {Lambrecht, Ellen and Bar{\'e}, Julie and Claeys, Myriam and Chavatte, Natascha and Bert, Wim and Sabbe, Koen and Houf, Kurt},
  issn         = {0736-6205},
  journal      = {BIOTECHNIQUES},
  keyword      = {ultrastructural analysis,intracellular survival,protozoan cysts,transmission electron microscopy,ACANTHAMOEBA-CASTELLANII,SURVIVAL,NEMATODA},
  language     = {eng},
  number       = {4},
  pages        = {181--188},
  title        = {Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa},
  url          = {http://dx.doi.org/10.2144/000114274},
  volume       = {58},
  year         = {2015},
}

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