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.
Highlights
The study of interactions between microorganisms and biotic or abiotic surfaces is fundamental to the field of microbial ecology, with the spatial detection and description of such interactions receiving much attention in recent years [1, 2]
We demonstrate the sensitivity of detection for specific bacteria with high spatial resolution on complex surface structures using scanning electron microscopy (SEM)
Gel electrophoresis was used to analyze the coupling of nanogold to oligonucleotide, as it should produce a significant shift in electrophoretic mobility [23]
Summary
The study of interactions between microorganisms and biotic or abiotic surfaces is fundamental to the field of microbial ecology, with the spatial detection and description of such interactions receiving much attention in recent years [1, 2]. Surface structures, such as redox-active mineral deposits or organic materials with sub-micron dimensions, are thought to strongly influence the ability of specific microorganisms to colonize and proliferate upon surfaces, which leads to the heterogeneous structure and composition of microbial communities [3, 4]. The application of FISH to complex environmental samples is often limited by the theoretical optical resolution of light microscopy (~0.25 μm) [7] and the background autofluorescence of certain type of samples, such as soil particles, tissues and mineral surfaces [8, 9].
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