Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 75, Issue 1, Pages 103-108Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2008.05.015
Keywords
FISH; CARD-FISH; tyramide signal amplification (TSA); stromatolite; mineralized biofilm
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Funding
- German Research Foundation [DFG-FOR 571
- AR 335/5]
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Modified protocols of fluorescence in situ hybridization (FISH) and catalyze reporter deposition fluorescence in situ hybridization (CARD-FISH) were developed in order to detect bacteria in situ in calcified stromatolite biofilms. Smooth, well-preserved thin sections of calcified biofilms (similar to 5 mu m thin, vertical sectioning of -1 cm deep) were obtained by cryo-sectioning using the adhesive tape-stabilization technique. A modified hybridization buffer was applied during hybridization to prevent calcite dissolution as well as false binding of oligonucleotide probes to the charged mineral surfaces. Particularly, bright and specific CARD-FISH signals allowed the detection of bacteria in intensively calcified biofilms even at low magnification, which is suitable for investigating millimeter- to centimeter-scale vertical distribution patterns of bacteria. (C) 2008 Elsevier B.V. All Fights reserved.
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