Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa
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...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2015-04-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000114274 |
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| author | Ellen Lambrecht Julie Baré Myriam Claeys Natascha Chavatte Wim Bert Koen Sabbe Kurt Houf |
| author_facet | Ellen Lambrecht Julie Baré Myriam Claeys Natascha Chavatte Wim Bert Koen Sabbe Kurt Houf |
| author_sort | Ellen Lambrecht |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e254f0df878f408e9f559c0c5d4ed700 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2015-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-e254f0df878f408e9f559c0c5d4ed7002025-08-20T02:25:50ZengTaylor & Francis GroupBioTechniques0736-62051940-98182015-04-0158418118810.2144/000114274Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoaEllen Lambrecht0Julie Baré1Myriam Claeys2Natascha Chavatte3Wim Bert4Koen Sabbe5Kurt Houf61Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium1Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium2Nematology Research Unit, Department of Biology, Ghent University, Ghent, Belgium1Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium2Nematology Research Unit, Department of Biology, Ghent University, Ghent, Belgium3Laboratory of Protistology and Aquatic Ecology, Department of Biology, Ghent University, Ghent, Belgium1Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Merelbeke, BelgiumCysts 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.https://www.future-science.com/doi/10.2144/000114274transmission electron microscopyprotozoan cystsultrastructural analysisintracellular survival |
| spellingShingle | Ellen Lambrecht Julie Baré Myriam Claeys Natascha Chavatte Wim Bert Koen Sabbe Kurt Houf Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa BioTechniques transmission electron microscopy protozoan cysts ultrastructural analysis intracellular survival |
| title | Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa |
| title_full | Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa |
| title_fullStr | Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa |
| title_full_unstemmed | Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa |
| title_short | Transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free-living protozoa |
| title_sort | transmission electron microscopy sample preparation protocols for the ultrastructural study of cysts of free living protozoa |
| topic | transmission electron microscopy protozoan cysts ultrastructural analysis intracellular survival |
| url | https://www.future-science.com/doi/10.2144/000114274 |
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