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

Ellen Lambrecht, Julie Baré, Myriam Claeys UGent, Natascha Chavatte, Wim Bert UGent, Koen Sabbe UGent and Kurt Houf UGent (2015) BIOTECHNIQUES. 58(4). p.181-188
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
ultrastructural analysis, intracellular survival, protozoan cysts, transmission electron microscopy, ACANTHAMOEBA-CASTELLANII, SURVIVAL, NEMATODA
journal title
BIOTECHNIQUES
Biotechniques
volume
58
issue
4
pages
181 - 188
Web of Science type
Article
Web of Science id
000352797800005
JCR category
BIOCHEMICAL RESEARCH METHODS
JCR impact factor
2.298 (2015)
JCR rank
41/77 (2015)
JCR quartile
3 (2015)
ISSN
0736-6205
DOI
10.2144/000114274
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
5938620
handle
http://hdl.handle.net/1854/LU-5938620
date created
2015-04-20 16:36:06
date last changed
2016-12-19 15:39:13
@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},
}

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.