Shaking and medium culture
Shaking culture The solid medium in the test tube or bottle is melted and inoculated, and then solidified. The simplest medium used is nutrient agar supplemented with 1% glucose. (1) Liquefy the medium and keep it at 100 ° C for 10 min to remove all dissolved oxygen. Cool to 45 ° C and use immediately. (2) Add the inoculum, rotate the test tube by hand to completely mix the contents of the test tube, and be careful not to generate bubbles. (3) Place the test tube in cold water to solidify the agar. (4) Incubate at the required temperature. Because the speed of oxygen penetrating the solid medium is very slow, the bottom of the test tube is very suitable for the growth of anaerobic bacteria. This method is also suitable for microaerobic bacteria and facultative anaerobic bacteria. Medium seal culture Liquid culture medium can be stored aseptically using sterile sealing materials. The test tube containing the liquid culture medium should be heated in a boiling water bath for 10 min to drive off dissolved oxygen, and then sterilized liquid petroleum jelly or paraffin is added to seal off the air. During inoculation, the petrolatum can be melted and inoculated with a capillary pipette (or the test tube is heated, inoculated immediately after cooling, and then sealed with a sterile sealing material I This method can be used to prove Clostridium perfringens in water or milk by rapid coagulation reaction. The medium used is anaerobic sterilized milk, which is sealed as described above. Vaspar is a very useful sealing substance, it is made by mixing and melting petroleum jelly and paraffin in the same proportion. After adding melted Vaspar to the medium and curing it, slowly heat the test tube with a flame In the Vaspar part, the solidified Vaspar is re-melted and tightly adhered to the wall of the test tube to isolate the air from the contact surface. Sterilized paraffin can also play a role in sealing the culture medium, but it can not detect the generation of gas, and will not dissolve some oxygen into the culture medium through the liquid paraffin sealing layer.
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