实验三土壤中细菌总数的测定.pptxVIP

会计学1实验三土壤中细菌总数的测定 第1页/共10页 第2页/共10页 第3页/共10页 2、The plate count (viable count) The number of bacteria in a given sample is usually too great to be counted directly. However, if the sample is serially diluted and then plated out on an agar surface in such a manner that single isolated bacteria form visible isolated colonies, the number of colonies can be used as a measure of the number of viable (living) cells in that known dilution. 第4页/共10页 Colony-Forming Units (CFUs) However, keep in mind that if the organism normally forms multiple cell arrangements, such as chains, the colony-forming unit may consist of a chain of bacteria rather than a single bacterium. In addition, some of the bacteria may be clumped together. Therefore, when doing the plate count technique, we generally say we are determining the number of Colony-Forming Units (CFUs) in that known dilution. By extrapolation, this number can in turn be used to calculate the number of CFUs in the original sample. 第5页/共10页 calculate the number of CFUs in the original samplenumber of CFUs per ml of sample = number of colonies (30-300 plate) X the dilution factor of the plate countedFor a more accurate count it is advisable to plate each dilution in duplicate or triplicate and then find an average count. 第6页/共10页 三 材料与器材无菌培养皿12套、1 mL无菌移液管10 支、土壤样品、天平等。第7页/共10页 四 操作步骤 无菌平皿编号→制备土壤稀释液→ 倾注平板→ 培养(48h) → 计数 第8页/共10页 五 注意事项倒平板时要注意控制培养基的温度。稀释操作时，要尽量减少样品稀释误差，而且每个稀释度的菌液应各用一支无菌吸管。为使菌落能在平板上均匀分布，样品液加入平皿后，应尽快倾注培养基并旋转混匀。 六 思考题要测定某种液体饮料中的活细菌总数，除了采用平板菌落计数法外，还可采用哪些方法？第9页/共10页