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Lab session 1 • Media preparation • Media sterilization • Use and care of the microscope • • • • • • Media – nutrients Used to grow microbes Glucose – energy Peptones – breakdown products of protein Good source of nitrogen, amino acids Yeast extract - vitamins • • • • • • Liquid medium – nutrient broth Large amount of culture Solid medium – nutrient agar Agar – solidifying agent Extracted from red algae Polysaccharide • Granules of agar from a commercial source • Add that to nutrient broth • Heat the mixture to dissolve granules • Solution cools down to 40oC agar becomes a solid Agar – good solidifying agent • • • • • • Microbes cannot breakdown agar Cannot metabolize No nutritional value Stays a solid even in incubators Study colony morphology Isolate a specific species from a sample • Agar – sterilized in the autoclave • Temperature 121oC, pressure 15psi, 15 minutes • Heat labile solutions – solutions of enzymes • Filtration • Microbes stay on the surface of filter • Solution free of microbes goes through the filter • • • • • Dry sterilization Glassware – pipets Oven – 180oC for 2 hours Chemical sterilization Disinfectant –clean table tops Microscope • • • • Use of oil immersion objective Focusing an object Total magnification Cleaning and storing