流式细胞计分析海洋微微型浮游生物:样品固定及贮存方法

以流式细胞计为海洋微微型浮游生物的分析工具,研究了醛类固定剂的固定、液氮贮存对于微微型浮游生物的细胞大小、密度、藻红素荧光值(FL2)和叶绿素荧光值(FL3)的影响.实验主要结论为1)液氮环境适于贮存固定的微微型浮游生物样品,而未固定样品在液氮中贮存后细胞密度有损失;2)醛类固定剂对微微型浮游生物样品的细胞密度基本没有影响;3)醛类固定剂可增强微微型浮游生物的荧光强度,其中戊二醛主要影响聚球藻的FL2强度,多聚甲醛主要影响FL3强度.认为多聚甲醛是适用于微微型浮游生物的固定剂,推荐使用浓度为0.8%~1.2%,此外戊二醛浓度为0.05%、多聚甲醛浓度为0.8%~1.2%的混合固定剂亦适用于固定微微型浮游生物.图4参15     

Picocyanobacteria – Sensitive Bioindicators of Contaminant Stress in an Alpine Lake (Traunsee Austria)

The abundance and photosynthetic activity ofpicocyanobacteria in the oligotrophic alpine lake Traunseewere measured at a station located close to the outlet ofindustrial soda waste and at a mid-lake reference stationduring spring, 1999 through to autumn, 2000.Picocyanobacterial numbers measured by flow cytometry inTraunsee (0.7–13.2 × 10⁴ ml^-1) were comparable tothose of other oligotrophic lakes, and there was nosignificant difference between the contaminated and thereference sampling location. Picoplankton (<2 m)photosynthetic rates measured in vitro by the ¹⁴C-technique were significantly reduced at the contaminated siterelative to the reference station at low photosyntheticallyavailable radiation (10 E m^-2 s^-1), while nodifference between these two stations was found at moderatelyhigh light intensity (100 E m^-2 s^-1). Theinvestigation was complemented by laboratory experiments withcultured picocyanobacteria. Three Synechococcus spp.strains were exposed to water taken from either of the twoTraunsee stations and from a control station located inneighbouring Attersee. Cell-specific photosynthetic activitymeasured by 4-h in vitro incubations revealed no significantdifference among the three stations investigated. Growthrates of the same three Synechococcus spp. strains weremeasured by flow cytometry over several days in thelaboratory. One strain, in particular, was sensitive to watertaken from the contaminated site; growth rate of this strainwas significantly reduced, relative to when exposed to watertaken from the reference station. Taken together, our resultsdemonstrate that picocyanobacteria are highly sensitivebioindicators of contaminant stress. The overall impact ofthe emissions from the industrial outlet on thepicocyanobacteria was, however, relatively minor.     

不同固定剂及保存时间对微微型浮游生物样品测定的影响

对比了用流式细胞术现场测定及用多聚甲醛、戊二醛及两者的混合物这三种常用固定剂进行固定保存对微微型浮游生物样品的影响。结果表明,在1个月内,细胞数量变化并不显著,但随着保存时间的增加,细胞数量明显减少,且散射光信号明显增大。除原绿球藻的藻红素荧光基本不变、微微型真核浮游植物的藻红素荧光大幅增加外,其余荧光信号均发生不同程度的减少。同时,戊二醛所引起的微微型真核浮游植物荧光信号的极大改变会影响类群的界定。如有可能,应该尽量现场测定微微型浮游生物样品,否则建议采用多聚甲醛或其与戊二醛的混合物作为固定剂,将样品保存于液氮中,待返回实验室后尽快进行测定。