1, culture should be based on the type of bacteria and purpose, such as selecting the culture method, medium, develop culture conditions (temperature, pH, time, the need for oxygen, etc.). The general procedure is to inoculate the specimen on solid medium and do the isolation culture. Then the resulting individual colonies are further identified by morphological, biochemical and serological reactions. Culture medium commonly used beef broth, peptone, sodium chloride, glucose, blood and other special substances required by certain bacteria prepared into liquid, semi-solid, solid and so on. General bacteria can be placed in aerobic conditions, 37 ℃ in 18 ~ 24 hours of growth. Anaerobic bacteria need to be placed in an anaerobic environment for 2 to 3 days after growth. Individual bacteria such as tuberculosis to culture as long as 1 month.
2, due to the ubiquity of bacteria, so from the preparation of the culture medium, the whole process of culture must be carried out in accordance with the requirements of aseptic operation, otherwise the external bacterial contamination of the specimen will lead to erroneous results; and cultivation of disease-causing organisms, once contaminated with the environment, it will cause cross-infection. For cultures performed for the purpose of disease diagnosis, appropriate specimens (blood, urine, stool, pus, secretions, etc.) should be selected, and the results obtained should be interpreted in the context of the clinical situation.
3. Take the method of photosynthetic bacterial culture as an example. The method of photosynthetic bacterial culture is sequentially divided into four steps: sterilization of containers and tools, preparation of culture medium, inoculation and culture management.
4. If the cultivated photosynthesizing bacteria are freshwater species, the strain culture can be prepared with distilled water, and the production culture can be prepared with sterilized tap water (or well water). If the cultivated photosynthesizing bacteria are seawater species, natural seawater can be used to prepare the medium, and it should be noted that when adding phosphorus into seawater, dipotassium hydrogen phosphate cannot be used, potassium dihydrogen phosphate should be used, otherwise a large amount of precipitation will be produced.
5, sterilization and disinfection of strains of culture medium should be used together with the culture container with autoclave sterilization pot sterilization. Small productive culture can be prepared culture solution with ordinary aluminum pot or large triangular flask boiling sterilization. Large-scale production culture is the precipitation of sand filtered water with bleach (or bleach) sterilization after use.
6, medium preparation according to the nutritional needs of the type of culture to choose the appropriate medium formula. According to the formula of the medium to the required substances weighed, one by one dissolved, mixed, formulated into a culture medium. Can also be prepared into the mother liquor, when used in proportion to add a certain amount can be.
7, inoculation of the medium is ready, should be inoculated immediately. Photosynthetic bacteria productive culture according to the amount of seed is relatively high, generally 20% to 50%, that is, the amount of strain mother liquor and the newly formulated culture solution although the ratio of 1:4 ~ 1:1, should not be less than 20%, especially microaerophilic culture, the inoculum amount should be higher, otherwise it is difficult to photosynthesizing bacteria in the culture solution of the absolute dominance of the culture, affecting the culture of the final yield and quality.
8, culture management of photosynthetic bacteria in the process of cultivation, management work, including daily management operations and testing, observation of growth, inspection and analysis of problems in three aspects.