Document Detail


Detection of prokaryotic cells with fluorescence in situ hybridization.
MedLine Citation:
PMID:  20809326     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fluorescence in situ hybridization with rRNA targeted oligonucleotide probes is nowadays one of the core techniques in microbial ecology, allowing the identification and quantification of microbial cells in environmental samples in situ. Next to the classic FISH protocol, which uses fluorescently monolabelled probes, the more sensitive CARD-FISH (also known as TSA-FISH), which involves an enzyme catalyzed signal amplification step, is becoming increasingly popular. This chapter describes protocols for both methods. While classic FISH has the advantage of being relatively cheap and easy to do on morphologically diverse samples, CARD-FISH offers a significantly higher sensitivity, allowing the detection of slow growing or metabolically inactive cells, which are below the detection limit of classic FISH. The drawback here is the considerably higher price for the probes and advanced cell fixation and permeabilization requirements that have to be optimized for different target cells.
Authors:
Katrin Zwirglmaier
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  659     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2010  
Date Detail:
Created Date:  2010-09-02     Completed Date:  2010-12-14     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  349-62     Citation Subset:  IM    
Affiliation:
British Antarctic Survey, Cambridge, UK. k.zwirglmaier@warwick.ac.uk
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MeSH Terms
Descriptor/Qualifier:
In Situ Hybridization, Fluorescence / methods*
Oligonucleotide Probes / genetics,  metabolism
Prokaryotic Cells / cytology*,  metabolism
Chemical
Reg. No./Substance:
0/Oligonucleotide Probes

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