微生物培养基制备
微生物培养基制备指通过混合营养物质、缓冲和维持渗透压平衡的试剂以及选择性抑制剂或指示剂来配制支持微生物生长和鉴别的琼脂或肉汤。这是微生物检测中腐败和致病微生物定期监测的例行任务。
培养基类型
微生物培养基应为所有或特定类型的微生物提供最佳的生长条件。其具体成分取决于培养物种和应用目标。培养基pH应根据微生物种类进行调节。微生物培养基根据多种参数分类,如化学成分、物理性质和功能。具体分类如下所述。
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按化学成分分类的微生物培养基
合成(成分明确)培养基是一种化学成分明确的培养基,常用于培养光能自养生物,如蓝细菌和光合原生生物,广泛应用于微生物代谢研究。
天然培养基是一种化学成分不确定的培养基,包括蛋白胨、肉类提取物和酵母提取物,可满足不同微生物的营养需求,主要用于培养营养需求复杂的苛求性菌。
按物理状态分类的微生物培养基
固体培养基使用1-7%琼脂或10-20%明胶凝固液体肉汤,用于分离不同微生物、获得纯菌落、制备斜面琼脂和穿刺琼脂培养基。
液体培养基不含任何凝固剂。接种和培养后,细胞形成可见小团或肉汤变混浊。
按制备方法分类的微生物培养基
即用型培养基是用培养板、培养瓶、试管或其他容器装载的液体或固体培养基,可直接使用,或可在重新融解和添加补充剂后直接使用。
商品化脱水合成培养基是一种干燥培养基,使用前需要加水和加工,配制成完全培养基或使用前添加补充剂的不完全培养基。
各别成分制备的培养基是完全由微生物实验室取各别成分配制的培养基。
按需培养基是由一种能够让高度浓缩的无菌培养基在实验室中维持提供的系统生产的。通过用无菌水稀释,它可根据需要生产所需量的无需高压灭菌即用型培养基。
微生物培养基制备
商品化脱水合成培养基应按照厂商提供的使用说明配制。调整蛋白胨、酵母提取物、琼脂、缓冲物质和抗生素这些基本成分的配方实现培养基的一致性。取所需量的脱水培养基或各别成分加入蒸馏水通过不断搅拌(必要时再进行加热)溶解,含琼脂的培养基在加热前应通过适当搅拌充分浸泡。必须调节pH,将培养基用适当容器分装进行高压蒸汽湿热灭菌。热敏感物质(如蛋白质、酶)应通过膜滤灭菌。
培养基应在规定温度储存防止成分发生变化,保存时间不得超过产品保质期。培养基必须无菌制备和保存,免遭微生物污染。通过密封包装和/或5 °C ± 3 °C储存尽量减少保存时的水分流失。避光、避热和脱水保存可延缓化学降解,以免造成氧化和抗菌剂损失等后果。
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