细胞培养板消毒只有紫外照射可以保证无菌吗?可不可以从清洗方面做点工作?
推荐回答（1个）
博凌科为解答：看你什么用途了，一般贴壁生长的细胞用重复利用的培养板效果不是很好，因为培养板表层在生产时涂有一层促进细胞贴壁的物质，在清洗后多半会失去。悬 浮或是半悬浮生长的细胞还可以。我们一般是先泡酸过夜，清洗干净，三蒸水清洗，再用无水酒精浸泡清洗，再在工作台里用无菌水过一遍，在盖上盖子在烘箱里烤干，待烤干后，打开在超净工作台里近紫外（距离紫外灯２０ｃｍ）照射半个小时以上，效果很好，没有污染过。其实不管老板有钱没钱，我们做实验室尤其是预实验或是摸条件时，我们一般都能省就省了，留点钱还不如买点好试剂呢：）我们一般是先清洗干净，泡酸过夜，三蒸水清洗，在盖上盖子在烘箱里烤干，待烤干后，有两种选择：1、在超净台里近紫外灯（距离紫外灯3０ｃｍ）照射半个小时以上， 六孔板一般半个小时，24孔板一个小时，96孔板时间更长，效果很好，没有污染过；2、到放射科照Co60，照完了尽快用。我们这里肿瘤细胞耐性很好，所以重复利用没什么问题，如果原代培养比较娇贵的细胞最好用新板子，也不是很贵。我们的经验是:清洗后用消毒液浸泡，泡酸过夜，自来水反复冲洗，双蒸水冲洗三遍，无尘环境晾干，用之前紫外灯照射半小时以上，效果很好。我们现在的 穷同仁一直在应用。永久了也会变黄，因此如果做mtt或比色时是不能用的，在不熟悉细胞培养的时候可以先学学细胞记数，熟练了用新的，不过肿瘤细胞可以在旧板子上长得 很好哟，原代的细胞比较难养，塑料的较好。紫外的穿透能力很弱，板子紫外照射时是否将盖子翻过来一起照射那？还有，细胞瓶紫外效果如何？板子泡酸后和瓶子一样的清洗方法清洗后，60度烤干，然后放在工作台里（打开盖子）用紫外线照射半小时以上，最好一小时或两小时，盖子同时反过来 照。瓶子照射没有效果，要用其他方法同意楼上大家的意见。96孔和24孔板子在做完了后要先泡清洗计，在用棉花搽洗每个孔，这样有些贴壁细胞才不会有细胞碎片的残留，在做mtt是很重 要，要不很不准的。后面就是冲洗，泡酸，3蒸水洗，紫外照射30－60分钟，不要时间太长，这样板子容易变色！但是如果是做MTT还是建议用新板子！旧板 子做，结果不准！泡酸时不要用刚配好的浓酸，因为那样会把培养板表面促进细胞贴附的一层膜腐蚀掉，细胞就无法贴壁了。用酸泡完后用自来水清洗，再用蒸馏水清洗3－5 次，然后泡在75％的酒精里（酒精用纯的无水乙醇），用前在紫外线下照射1h。基本可保证95％以上无菌，用这些板做做预实验是没问题的。1.钴60照射适合大量的板同时灭菌，最好攒一箱，再送去灭菌。2.紫外消毒适合少量的培养皿或培养板，比如培养板，将盖打开，翻过来，连同板一起在紫外下照射2小时即可，事先不用酒精泡。
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