Format

Send to

Choose Destination
See comment in PubMed Commons below
Appl Environ Microbiol. 2005 Nov;71(11):7321-6.

Catalyzed reported deposition-fluorescence in situ hybridization protocol to evaluate phagotrophy in mixotrophic protists.

Author information

1
Unitat de Limnologia, Departament de Biogeoquimica Aquàtica, Centre d'Estudis Avançats de Blanes-CSIC, C/Accés a la Cala St. Francesc, 14, E-17300 Blanes (Girona), Spain. jmmedina@ceab.csic.es

Abstract

We describe a catalyzed reported deposition-fluorescence in situ hybridization (CARD-FISH) protocol particularly suited to assess the phagotrophy of mixotrophic protists on prokaryotes, since it maintains cell and plastid integrity, avoids cell loss and egestion of prey, and allows visualization of labeled prey against plastid autofluorescence. This protocol, which includes steps such as Lugol's-formaldehyde-thiosulfate fixation, agarose cell attachment, cell wall permeabilization with lysozyme plus achromopeptidase, and signal amplification with Alexa-Fluor 488, allowed us to detect almost 100% of planktonic prokaryotes (Bacteria and Archaea) and, for the first time, to show archaeal cells ingested by mixotrophic protists.

PMID:
16269774
PMCID:
PMC1287680
DOI:
10.1128/AEM.71.11.7321-7326.2005
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Support Center