Vaulot D, Courties C, Partensky F
CNRS, Station Biologique, Roscoff, France.
Cytometry. 1989 Sep;10(5):629-35. doi: 10.1002/cyto.990100519.
A simple method was developed to preserve marine phytoplankton populations so that delayed flow cytometric analyses could be performed. The method consisted of immediate fixation with 1% glutaraldehyde (final concentration) followed by storage in liquid nitrogen. The method was tested on individual algal species and on natural samples from both coastal and pelagic waters. In most cases, it caused little cell loss and preserved well both forward angle light scatter and chlorophyll fluorescence, but phycoerythrin fluorescence sometimes was significantly increased. The technique performed best for the small-sized picoplankton (below 2 microns) such as Synechococcus cyanobacteria or the newly discovered oceanic prochlorophytes. For larger-sized cells it had to be applied on a case by case basis as some fragile species, particularly dinoflagellates and cryptophytes, were poorly preserved.
开发了一种简单的方法来保存海洋浮游植物种群,以便能够进行延迟的流式细胞术分析。该方法包括立即用1%戊二醛(终浓度)固定,然后储存在液氮中。该方法在单个藻类物种以及来自沿海水域和远洋水域的天然样本上进行了测试。在大多数情况下,它几乎不会导致细胞损失,并且能很好地保存前向角光散射和叶绿素荧光,但藻红蛋白荧光有时会显著增加。该技术对小型微微型浮游生物(小于2微米),如蓝藻聚球藻或新发现的海洋原绿藻效果最佳。对于较大尺寸的细胞,必须根据具体情况应用,因为一些脆弱的物种,特别是甲藻和隐藻,保存效果不佳。