111Indium labeling of microorganisms to facilitate the investigation of bacterial adhesion

The ability of bacteria to adhere to polymeric interfaces has attracted considerable attention in recent years. Metabolic labeling of microorganisms with 35S-methionine or other beta-emitters is commonly utilized for quantification of bacterial adhesion to biopolymers. Since the use of these isotopes is cumbersome, the possibility of labeling the microorganisms with 111Indium, a strong gamma-emitter, was explored. This report demonstrates that bacteria can be easily labeled with 111Indium. Staphylococcus aureus, Staphylococcus epidermiids, and Pseudomonas aeruginosa were labeled with either 111Indium-oxine or 35S-methionine; and labeling efficiency, retention of incorporated labels, and growth kinetics of labeled bacteria were compared under identical experimental conditions. Bacteria labeled with 111In-oxine incorporated approximately 90% of radioactivity within 10 min, whereas 35S-methionine incorporation required many hours of incubation. Both the incorporated isotopes were gradually released by rapidly growing bacteria into the suspension medium. Of the total incorporated labels, approximately 20% 111In and 15% 35S were released in the surrounding medium every 24 h. No release of incorporated labels occurred when cells were fixed with 2.5% buffered glutaraldehyde. Growth kinetics and scanning or transmission electron microscopic analysis showed no detectable differences among control (nonlabeled), 111In-, or 35S-labeled bacteria. Labeling of bacteria with 111In-oxine does not interfere with bacterial adherence. These observations suggest that 111In incorporation provides a simple and rapid method of labeling of microorganisms. Compared to currently available techniques, the use of 111In-labeled bacteria will facilitate the quantitation of adherent bacteria to interfaces.