AIMS: To develop a scanning electron microscopic approach using in situ hybridization (SEM-ISH) for gaining both genetic and morphological information about target bacteria. METHODS AND RESULTS: Target cells were hybridized with DNA-targeted polynucleotide probes, and a tyramide signal amplification system was used to increase the sensitivity. The protocol of SEM-ISH enabled to detect low copy number target DNA sequences in individual cells. CONCLUSIONS: SEM-ISH allowed the in situ detection of bacteria carrying a specific gene. SIGNIFICANCE AND IMPACT OF THE STUDY: Combining morphological study with SEM and ISH techniques appears to be a valuable tool to understand the spatial distribution of target cells in complex microbial communities on various materials.
AIMS: To develop a scanning electron microscopic approach using in situ hybridization (SEM-ISH) for gaining both genetic and morphological information about target bacteria. METHODS AND RESULTS: Target cells were hybridized with DNA-targeted polynucleotide probes, and a tyramide signal amplification system was used to increase the sensitivity. The protocol of SEM-ISH enabled to detect low copy number target DNA sequences in individual cells. CONCLUSIONS: SEM-ISH allowed the in situ detection of bacteria carrying a specific gene. SIGNIFICANCE AND IMPACT OF THE STUDY: Combining morphological study with SEM and ISH techniques appears to be a valuable tool to understand the spatial distribution of target cells in complex microbial communities on various materials.