A rapid workflow for the characterization of small numbers of unicellular eukaryotes by using correlative light and electron microscopy
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
32169550
DOI
10.1016/j.mimet.2020.105888
PII: S0167-7012(20)30048-8
Knihovny.cz E-resources
- Keywords
- CLEM, Cyst, EDX, Eukaryotic microorganisms, FESEM, FIB/SEM, Giardia,
- MeSH
- Microscopy, Electron methods MeSH
- Eukaryota classification isolation & purification ultrastructure MeSH
- Phylogeny MeSH
- Microscopy, Electron, Scanning methods MeSH
- Microscopy methods MeSH
- Workflow MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic parasite. Correlative light and electron microscopy (CLEM) allows for the selection of individual cells and is economical in terms of time and cost. We describe a cyst purification protocol in combination with an adhesive coating for fixation and ultrathin embedding that results in excellent preservation of cell morphology. The application of advanced structural and analytical EM methods, such as high-resolution field emission scanning electron microscopy (FESEM), focused ion beam tomography (FIB/SEM), and energy-dispersive X-ray spectroscopy (EDX) analysis, is demonstrated. The workflow represents a new approach for studying the cellular and organelle architecture of rare and "difficult to culture" microorganisms.
References provided by Crossref.org
The Giardia ventrolateral flange is a lamellar membrane protrusion that supports attachment
