title: Polygold-FISH for signal amplification of metallo-labeled microbial cells.
abstract: An improved in situ hybridization approach (Polygold-FISH) using biotinylated probes targeting multiple locations of the 16 S ribosomal subunit, followed by fluoronanogold-streptavidin labeling and autometallographic enhancement of nanogold particles was developed as a means of signal amplification of metallo-labeled cells, without the need for Catalyzed Reporter Deposition (CARD). Bacterial cells were readily detected based on their gold-particle signal using scanning-electron microscopy and energy-dispersive X-ray spectroscopy when contrasted with controls or cells hybridized with a single probe. Polygold-FISH presents an alternative to CARD-FISH, circumventing the need for aggressive oxidants, which is useful when products of microbial respiration such as those relevant at the microbe-mineral interface could be altered during processing for visualization.
authors: Department of Civil and Environmental Engineering, Colorado School of Mines, Golden, CO, USA.
authors: Robert
authors: R
authors: Almstrand
authors: Department of Civil and Environmental Engineering, Colorado School of Mines, Golden, CO, USA.
authors: Dina M
authors: DM
authors: Drennan
authors: Department of Civil and Environmental Engineering, Colorado School of Mines, Golden, CO, USA.
authors: Jonathan O
authors: JO
authors: Sharp
mesh_terms: D001419:Bacteria / Q000166:cytology / Q000235:genetics* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D053768:Metal Nanoparticles; D020592:Shewanella / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D020614:Shewanella putrefaciens / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure
keywords: Electron microscopy; FISH; Gold; Metallo-labeling