A Correlative Light-Electron Microscopy (CLEM) Protocol for the Identification of Bacteria in Animal Tissue, Exemplified by Methanotrophic Symbionts of Deep-Sea Mussels
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Bacterial symbionts associated with animal tissues play major roles in the functioning of various ecosystems. Identification of bacteria often relies on marker gene comparative sequence analysis and fluorescence in situ hybridization (FISH). However, analysis of bacteria and host ultrastructure using transmission electron microscopy (TEM) can be equally important to understand the localization of bacteria and the degree of host-symbiont integration. We here provide a protocol which allows both FISH and TEM to be performed sequentially on a single section of tissue. Observations can then be superimposed, allowing ultrastructural investigation to be coupled with proper FISH-based identification of bacteria.
Keywords:Correlative microscopy Fluorescence in situ hybridization Symbiosis Transmission electron microscopy
- 7.Abramoff MD, Magalhaes PJ, Ram SJ (2004) Image processing with ImageJ. Biophoton Int 11:36–42Google Scholar
- 8.Duperron S (2015) Characterization of bacterial symbionts in deep-sea metazoans: protocols for conditioning, fluorescence in situ hybridization, and image analysis. In: Mc Gentity TJ, Timmis KN, Nogales B (eds) Hydrocarbons and lipid microbiology protocols. Springer. doi: 10.1007/8623_2015_73
- 9.Duperron S, Halary S, Lorion J et al (2008) Unexpected co-occurrence of 6 bacterial symbionts in the gill of the cold seep mussel Idas sp. (Bivalvia: Mytilidae). Environ Microbiol 10:433–445Google Scholar