Technical Reference #1846

1846.

Targeted Delivery of a Photosensitizer to Aggregatibacter actinomycetemcomitans Biofilm Peter Suci; Sebyung Kang; Rudolf Gmu¨r; Trevor Douglas; and Mark Young, Montana State University, Antimicrobial Agents and Chemotherapy, 54(1846), (2010)
Link To Paper

Materials & Methods:
Bacterial strain and culture conditions. Aggregatibacter actinomycetemcomitans (formerly Actinobacillus actinomycetemcomitans) strain D7 a rough colony clinical isolate obtained from the central incisor of an African-American female patient with generalized aggressive periodontitis was provided by Casey Chen University of Southern California. The liquid medium was modified tryptic soy broth (MTSB) consisting of (per liter) 30 g tryptic soy broth and 3 g yeast extract. The solid medium was MTSB with 50 ml fetal bovine serum (HyClone) and 15 g Bacto agar added (per liter). Frozen stocks were maintained at 80°C in 20% glycerol and 80% MTSB. Biofilms were cultured in 96-well microtiter plates with glass bottoms (P96G-1.5-5-F; MatTek Corp.) to enable confocal laser scanning microscope (CLSM) images to be acquired. The biofilm inoculum was prepared by looping single colonies cultured on solid medium at 37°C in 5% CO2 for 72 h into a 1-ml aliquot of liquid medium and dispersing the suspension of cell aggregates by repeated pipetting followed by vortexing and finally diluting the suspension to obtain approximately 107 CFU/ml. No attempt was made to isolate single cells from cell aggregates as described by Kaplan et al. (27). Biofilms were cultured at 37°C in 5% CO2. Medium was replaced at 24 h for biofilms cultured for 48 h.

Microscopic Technique
Confocal laser scanning microscope

Cell Type(s)
Aggregatibacter actinomycetemcomitans