title: High-resolution and specific detection of bacteria on complex surfaces using nanoparticle probes and electron microscopy.
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.
authors.affiliation: Centre for Marine Bio-Innovation, School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
authors.forename: Jun
authors.initials: J
authors.lastname: Ye
authors.affiliation: Centre for Marine Bio-Innovation, School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
authors.forename: Shaun
authors.initials: S
authors.lastname: Nielsen
authors.affiliation: School of Materials Science and Engineering, The University of New South Wales, Sydney, Australia.
authors.forename: Stephen
authors.initials: S
authors.lastname: Joseph
authors.affiliation: Centre for Marine Bio-Innovation, School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
authors.forename: Torsten
authors.initials: T
authors.lastname: Thomas
mesh_terms: D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D001431:Bacteriological Techniques / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D006046:Gold / Q000737:chemistry; D017403:In Situ Hybridization / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D042808:Neisseria sicca / Q000235:genetics / Q000302:isolation & purification; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012680:Sensitivity and Specificity; D013052:Spectrometry, X-Ray Emission
chemical_list: D015345:Oligonucleotide Probes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D006046:Gold