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.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Lambrecht E,Baré J,Claeys M,Chavatte N,Bert W,Sabbe K,Houf Kdoi
10.2144/000114274subject
Has Abstractpub_date
2015-04-01 00:00:00pages
181-8issue
4eissn
0736-6205issn
1940-9818pii
000114274journal_volume
58pub_type
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