A new method of bacterial identification using gold immunolabelling and scanning electron microscopy

This study sought to develop an immunolabelling technique to identify specific bacteria by scanning electron microscopy. Bacterial suspensions were prepared of Actinobacillus actinomycetemcomitans, Porphyromonas (Bacteroides) gingivalis and mixtures of these species with other genera of common oral microorganisms. This method was also used to examine subgingival plaques from 6 subjects with chronic adult periodontitis. Sample preparation consisted of prefixation of the bacterial suspensions with 0.2% glutaraldehyde, incubation with species-specific rabbit antisera and goat anti-rabbit IgG conjugated with colloidal gold particles, postfixation in 2% glutaraldehyde and dehydration in ethanol. Finally, the samples were dried, coated with evaporated carbon and examined by scanning electron microscopy. Pure cultures and artificial mixtures of A. actinomycetemcomitans and Bact. gingivalis were specifically labelled by gold probes as demonstrated by both secondary and back-scattered imaging. These species were also evident in samples of subgingival plaque. The findings indicate that this new technique can be used to identify specific microorganisms both in plaque samples and on the root surface of extracted teeth.