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If the Pseudomonas biofilm is grown in a liquid growth medium that lacks a Carbon source, the bacteria will turn to the styrofoam as a carbon source and begin to degrade it
1. Select one of the square petri plates containing the broth.
2. Onto the surface of the agar, aseptically place one sheet of sterile filter paper. Sterilize a pair of forceps by dipping them in a container of alcohol and flaming them.Place the sheet so that no air bubbles are trapped under the filter paper.
3. Dilute the overnight culture. Thoroughly mix the culture by vortexing or by rolling the tube containing the culture between the palms of your hands.
4. Uniformly moisten the filter paper with 1 ml of the diluted bacterial culture.
5. Select a clean 1 X 3 inch microscope slide and using the forceps, immerse the slide in a container of alcohol and sterilize it by flaming. Then carefully lay the slide on top of the filter paper. make sure there are no air bubbles
6. Repeat step 5 three more times.
7. Between Periods 1 and 2 If possible, remoisten the plate after 24 hours with 1 ml of the culture.
Period 1
1. After two days the biofilms may be harvested by carefully lifting the slides from the filter paper surface with a pair of sterile forceps (flamed). The biofilm should appear as a slimy layer coating the under surface of the slide. Lifting too rapidly may disturb the biofilm causing large sections to slough off the slide.