Overnight cultures were grown by adding a colony from a freshly streaked plate of A. baumannii BAA-1605 to a test tube containing 3 mL of tryptic soy broth (TSB) (Fisher Scientific, DF0370-17-3). This was incubated at 37°C overnight with shaking (120 rpm). The culture was diluted to OD600 = 0.5 and 2.5 μL spotted on the center of a sterile 19-mm polycarbonate membrane (Whatman Nuclepore polycarbonate hydrophilic membranes, Cytiva #10417004). Membranes were sterilized by exposure to UV for 10 min each side and placed (shiny side up) on tryptic soy agar (TSA) plates before spotting. Biofilms were grown on the membranes by incubation at 37°C for 24 hours.
Nuclepore polycarbonate hydrophilic membranes
Manufactured by Cytiva
Nuclepore polycarbonate hydrophilic membranes are microporous membranes made of polycarbonate material. They feature a uniform pore structure and are designed to be hydrophilic, allowing for efficient filtration of aqueous solutions.
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Lab products found in correlation
2 protocols using nuclepore polycarbonate hydrophilic membranes
1
Biofilm Formation of A. baumannii
2
In Vitro Agar Biofilm Protocol
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For the in vitro agar membrane biofilm model, a generation 1 streak plate was prepared by streaking frozen stock culture on a tryptic soy agar (TSA) plate. The generation 1 plate was incubated at 37°C for 24 hours. A generation 2 plate was made by streaking a colony picked from the generation 1 plate on a fresh TSA plate. The generation 2 plate was incubated at 37°C for 24 hours. A bacterial broth was made by adding a single colony from the generation 2 plate to a 2 mL TSB solution. The broth culture was placed in the incubator at 37°C at 150 rpm until it reached a turbidity of 0.5 McFarland (approximately 2–4 hours). Finally, 2.5 μL of this broth was spotted on the center of a UV-sterilized 13 mm polycarbonate membrane (Whatman Nuclepore polycarbonate hydrophilic membranes, Cytiva #10417001) atop a TSA plate. Biofilms were grown on the membranes for 24 hours at 37°C.
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