用光谱流式细胞术分析铜绿微囊藻的生理:化学和光照的影响

Emma T. Brentjens, Elizabeth A. K. Beall, Robert M. Zucker
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引用次数: 0

摘要

M。使用Cytek Aurora光谱流式细胞仪观察铜绿假单胞菌的荧光变化,该流式细胞仪包含5个激光器和64个位于365至829 nm之间的窄带检测器。用不同浓度的h2o2处理蓝藻,并在接触后1 ~ 8天进行监测。在652-669 nm的检测器上测量了黄绿色激光(550 nm)激发蓝藻产生的红色荧光,在532-550 nm的检测器上测量了紫色激光(405 nm)激发蓝藻产生的绿色荧光。在加入h2o2后,测量了这些参数的变化。红色荧光强度在24小时开始增加。随后,红色荧光强度每天都在下降。相比之下,绿色荧光在24小时增加,并在8天的研究期间保持高于对照组。与h2o2对M的荧光强度效应相似。暴露于乙酰丙酮、二脲(DCMU)、过氧乙酸和色氨酸后观察绿脓杆菌的荧光发射。在暴露于h2o2 24天期间,也观察到在h2o2处理的蓝藻中生长最小。在另一项实验中,将经过h2o2处理的蓝藻暴露在高强度蓝光(14 mW)和紫外线(1 mW)光下,以评估光胁迫对荧光发射的影响。蓝光、紫外光与h2o2的组合对M有增效作用。与单独暴露于任何一种刺激相比,铜绿菌在对照和处理样品之间引起更大的荧光差异。这些实验表明,红色和绿色荧光的早期增加可能是由于光合作用处理光子的能力受到抑制。进一步研究驱动这些荧光增加的机制是必要的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Analysis of Microcystis aeruginosa physiology by spectral flow cytometry: Impact of chemical and light exposure
M . aeruginosa fluorescent changes were observed using a Cytek Aurora spectral flow cytometer that contains 5 lasers and 64 narrow band detectors located between 365 and 829 nm. Cyanobacteria were treated with different concentrations of H 2 O 2 and then monitored after exposure between 1 and 8 days. The red fluorescence emission derived from the excitation of cyanobacteria with a yellow green laser (550 nm) was measured in the 652–669 nm detector while green fluorescence from excitation with a violet laser (405 nm) was measured in the 532–550 nm detector. The changes in these parameters were measured after the addition of H 2 O 2 . There was an initial increase in red fluorescence intensity at 24 hours. This was followed by a daily decrease in red fluorescence intensity. In contrast, green fluorescence increased at 24 hours and remained higher than the control for the duration of the 8-day study. A similar fluorescence intensity effect as H 2 O 2 on M . aeruginosa fluorescence emissions was observed after exposure to acetylacetone, diuron (DCMU), peracetic acid, and tryptoline. Minimal growth was also observed in H 2 O 2 treated cyanobacteria during exposure of H 2 O 2 for 24 days. In another experiment, H 2 O 2 -treated cyanobacteria were exposed to high-intensity blue (14 mW) and UV (1 mW) lights to assess the effects of light stress on fluorescence emissions. The combination of blue and UV light with H 2 O 2 had a synergistic effect on M . aeruginosa that induced greater fluorescent differences between control and treated samples than exposure to either stimulus individually. These experiments suggest that the early increase in red and green fluorescence may be due to an inhibition in the ability of photosynthesis to process photons. Further research into the mechanisms driving these increases in fluorescence is necessary.
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