Coupling bacterial activity measurements with cell sorting by flow cytometry

被引:74
作者
Servais, P
Courties, C
Lebaron, P
Troussellier, M
机构
[1] Free Univ Brussels, Grp Microbiol Milieux Aquat, B-1050 Brussels, Belgium
[2] UPMC, Lab Arago, CNRS UMR 7628, F-66651 Banyuls sur Mer, France
[3] UPMC, Lab Arago, CNRS UMR 7621, F-66651 Banyuls sur Mer, France
[4] Univ Montpellier 2, UMR CNRS 5556, Lab Hydrobiol Marine, F-34095 Montpellier 05, France
关键词
D O I
10.1007/s002489900160
中图分类号
Q14 [生态学(生物生态学)];
学科分类号
071012 ; 0713 ;
摘要
A new procedure to investigate the relationship between bacterial cell size and activity at the cellular level has been developed; it is based on the coupling of radioactive labeling of bacterial cells and cell sorting by flow cytometry after SYTO 13 staining. Before sorting, bacterial cells were incubated in the presence of tritiated leucine using a procedure similar to that used for measuring bacterial production by leucine incorporation and then stained with SYTO 13. Subpopulations of bacterial cells were sorted according to their average right-angle light scatter (RALS) and fluorescence. Average RALS was shown to be significantly related to the average biovolume. Experiments were performed on samples collected at different times in a Mediterranean seawater mesocosm enriched with nitrogen and phosphorus. At four sampling times, bacteria were sorted in two subpopulations (cells smaller and larger than 0.25 mu m(3)). The results indicate that, at each sampling time, the growth rate of larger cells was higher than that of smaller cells. In order to confirm this tendency, cell sorting was performed on six subpopulations differing in average biovolume during the mesocosm follow-up. A clear increase of the bacterial growth rates was observed with increasing cell size for the conditions mel in this enriched mesocosm.
引用
收藏
页码:180 / 189
页数:10
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