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      常用培養基的制備,滅菌與消毒實驗

      發布時間:

      2022-12-27

      作者:

      即用型干粉培養基


      1,實驗目的: 
      了解培養基的配制原理;掌握配制培養基的一般方法和步驟;了解常見滅菌,清毒基本原理及方法,掌握干熱天菌,高壓蒸汽滅菌及過濾除菌的操作方法。 
       
      2,實驗原理: 
      培養基是人工按一定比例配制的供微生物生長繁殖和合成代謝產物所需要的營養物質的混合物。培養基的原材料可分為碳源,氮源,無機鹽,生長因素和水。根據微生物的種類和實驗目的不同,培養基也有不同的種類和配制方法。 
      牛肉膏蛋白胨培養基是一種應用最廣泛和最普通的細菌基礎培養基,有時又稱為普通培養基。由于這種培養基中含有一般細胞生長繁殖所需要的最基本的營養物質,所以可供微生物生長繁殖之用。 
      干熱天菌,高壓蒸汽滅菌方法主要是通過升溫使蛋白質變性從而達到殺死微生物的效果。 
       
      3,試劑與器材: 
      1.器材 試管,三角瓶,燒杯,量筒,玻璃棒,培養基,分裝器,天平,牛角匙,高壓蒸汽滅菌鍋,pH度紙,棉花,牛皮紙,記號筆,麻繩,紗布,吸管,培養皿,電烘箱,注射器,微孔濾膜過濾器,鑷子等。 
      2.試劑 牛肉膏,蛋白胨,NaCl,瓊脂。 
       
      4,實驗內容: 
      1.稱量—溶化—調pH—過濾—分裝—加塞—包扎—滅菌—無菌檢查。 
      2.干熱滅菌:裝入待滅菌物品—升溫—恒溫—降溫—開箱取物。 培養基的滅菌圖_青島日水
      3.高壓蒸汽滅菌:加水—裝物品—加蓋—加熱—排冷空氣—加壓—恒壓—降壓回零—排汽—取物—無菌檢查。 
      4.過濾除菌:組裝滅菌—連接—壓濾—無菌檢查—清洗滅菌。 
       
      5,關鍵步驟及注意事項: 
      1.要嚴格按配方配制。 
      2.調pH不要過頭。 
      3.干熱滅菌要注意物品不要堆放過緊,注意溫度的時間控制,70ºC以下放物,取物。 
      4.高壓滅菌要注意物品不要過多,加熱后排除冷空氣,到時降壓回零取物。 
      5.過濾除菌要注意各部件滅菌,壓濾時,壓力要適當,不可太猛太快,濾膜要注意清洗保存。 

      Preparation, sterilization and disinfection of commonly used medium
      1, the purpose of the experiment is:
      Understand the preparation principle of the medium, master the general methods and steps of preparing the medium, understand the basic principles and methods of common sterilization and detoxification, master the operating methods of dry and hot days, high-pressure steam sterilization and filtration sterilization.
      2, the experimental principle:
      Medium is a mixture of nutrients for microbial growth, reproduction and metabolites. The raw materials of the culture medium can be divided into carbon source, nitrogen source, inorganic salt, growth factor and water. According to the different types of microorganisms and experimental purposes, there are different kinds of media and preparation methods.
      Beef extract peptone culture medium is one of the most widely used and common bacterial base medium, sometimes known as ordinary medium. Because this kind of medium contains the most basic nutrients for the growth and reproduction of ordinary cells, it can be used for the growth and reproduction of microorganisms.
      Dry-hot days bacteria, high-pressure steam sterilization method is mainly through heating protein denaturation to achieve the effect of killing microorganisms.
      3, reagents and equipment:
      1. Equipment test tube, triangle bottle, beaker, measuring barrel, glass rod, culture medium, sub-packer, balance, horn spoon, high-pressure steam sterilization pot, pH paper, cotton, kraft paper, marker pen, hemp rope, gauze, straw, culture dish, electric oven, syringe, microporous filter, tweezers, etc.
      2. reagent beef paste, peptone, NaCl, agar.
      4, experimental contents:
      1. weighing - dissolving - adjusting pH - Filtration - packing - plus - dressing - sterilization - sterility test.
      2. dry heat sterilization: loading the product to be sterilized - warming up - constant temperature - cooling - opening the box.
      3. Sterilization by high pressure steam: adding water, loading, covering, heating, discharging cold air, pressurizing, constant pressure, depressurizing back to zero, discharging steam, extracting, sterilization inspection.
      4. filtration sterilization: assembly sterilization - connection - pressure filtration - sterility test - cleaning and sterilization.
      5, the key steps and matters needing attention:
      1., we must strictly prepare the formula.
      2. don't overdo pH.
      3. Dry-heat sterilization should pay attention to the items do not stack too tight, pay attention to the time control of temperature, 70 C below the place, take things.
      4. High-pressure sterilization should pay attention to not too many articles, after heating, cold air is removed, when the time comes to reduce the pressure back to zero.
      5. Pay attention to the sterilization of each component in filtration, pressure should be appropriate, not too violent and too fast, filter membrane should pay attention to cleaning and preservation.
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