High-resolution and specific detection of bacteria on complex surfaces using nanoparticle probes and electron microscopy

Jun Ye; Shaun Nielsen; Stephen Joseph; Torsten Thomas

doi:10.1371/journal.pone.0126404

High-resolution and specific detection of bacteria on complex surfaces using nanoparticle probes and electron microscopy

PLoS One. 2015 May 27;10(5):e0126404. doi: 10.1371/journal.pone.0126404. eCollection 2015.

Authors

Jun Ye¹, Shaun Nielsen¹, Stephen Joseph², Torsten Thomas¹

Affiliations

¹ Centre for Marine Bio-Innovation, School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
² School of Materials Science and Engineering, The University of New South Wales, Sydney, Australia.

Abstract

The study of the interaction of bacteria with surfaces requires the detection of specific bacterial groups with high spatial resolution. Here, we describe a method to rapidly and efficiently add nanogold particles to oligonucleotide probes, which target bacterial ribosomal RNA. These nanogold-labeled probes are then used in an in situ hybridization procedure that ensures both cellular integrity and high specificity. Electron microscopy subsequently enables the visualization of specific cells with high local precision on complex surface structures. This method will contribute to an increased understanding of how bacteria interact with surface structures on a sub-micron scale.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacteria / genetics*
Bacteria / isolation & purification
Bacteriological Techniques / instrumentation
Bacteriological Techniques / methods*
Escherichia coli / genetics
Escherichia coli / isolation & purification
Gold / chemistry
In Situ Hybridization / methods
Microscopy, Electron, Scanning / methods*
Nanoparticles / chemistry*
Neisseria sicca / genetics
Neisseria sicca / isolation & purification
Oligonucleotide Probes / chemical synthesis
Oligonucleotide Probes / chemistry
Oligonucleotide Probes / genetics
RNA, Bacterial
RNA, Ribosomal
Sensitivity and Specificity
Spectrometry, X-Ray Emission

Substances

Oligonucleotide Probes
RNA, Bacterial
RNA, Ribosomal
Gold

Grants and funding

The study was supported by the Australian Research Council. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.