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用室内培养的塔玛亚历山大藻(Alexandrium tamarense ATHK藻株)作为实验对象,分别选择了5%福尔马林固定后常温保存、95%乙醇固定后常温保存、-20℃冷冻保存、鲁哥氏液(Lugols solution)固定后常温保存等方法,在5,15,30,60 d后对保存的样品进行单细胞PCR反应,并与新鲜样品进行比较。实验结果表明,用95%乙醇保存、-20℃冷冻保存、鲁哥氏液保存的样品在60 d后仍可扩增出目标条带,与新鲜样品没有差别,但是样品中的藻细胞形态有一些改变;而以5%福尔马林固定保存的样品只能在5 d后扩增出目标条带,但藻细胞形态比较完好。因此,对用于单细胞PCR实验的微藻样品,短期内(不多于5 d)可以采用福尔马林保存,而需要长期保存的样品,应当采用乙醇或鲁哥氏液固定,或者采用-20℃冷冻保存的方法。
Using Alexandrium tamarense ATHK strain cultured indoors, we selected 5% formalin fixed at room temperature, fixed at 95% ethanol and stored at -20 ℃, After the Lugols solution was fixed and stored at room temperature, the stored samples were subjected to single-cell PCR after 5, 15, 30 and 60 days, and compared with fresh samples. The experimental results show that the samples preserved in the culture medium of Ruoerguosta still retain their target bands after being stored in 95% ethanol at -20 ℃ for about 60 days, which is similar to that of the fresh samples. However, Some changes were observed. However, the samples stained with 5% formalin only amplified the target bands after 5 days, but the morphology of algal cells was intact. Therefore, the microalgae samples used for single-cell PCR experiments can be stored in short-term (not more than 5 days) using formalin, and the samples requiring long-term preservation should be fixed with ethanol or Rugal’s solution, -20 ℃ cryopreservation method.