基于Nano SIMS表征荧蒽在单体微生物体内运移研究的实验数据处理方法
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摘要
利用纳米二次离子技术(Nano SIMS)对荧蒽在盐单胞菌BDG-3体内的代谢过程进行观测,对选中区域中质量数为12、13、16、26的信号进行收集;并结合Image J和Python软件对所获得的数据进行分析.结果表明:菌体内~(12)C~-与~(13)C~-呈正相关,相关系数极强,为0.998.~(16)O~-与~(26)(~(12)C~(14)N)~-呈正相关,相关系数较强,为0.813.并且,通过构建雷达图对不同元素进行综合性评价,计算了不同时间点下的假单胞菌BDG-3平均面积、平均周长、协调性Vi.结果表明在3h时,菌体的平均面积最大,达到1.39,3h后又迅速下降,6h达到最低为0.003 1,评价函数f(S_i,V_i)显示加入荧蒽3h左右,盐单胞菌BDG-3生长状态最佳.本研究结合Python与Image J软件,建立了一套完整、直观的微生物Nano SIMS图像处理与评价的方法,从而为研究在微生物体内更为复杂的环境污染物质的降解代谢过程提供了全新、可靠的分析手段.
Nano-secondary ion technique(Nano SIMS)was used to investigate fluoranthene metabolism in Halomonassp.BDG-3.Mass signals at 12,13,16,and 26 were collected and analyzed with Image J and Python.~(12)C~- and ~(13)C~- were found to be positively correlated,with a strong correlation coefficient of 0.998.16 O-was found to be positively correlated with ~(26)(~(12)C~(14)N)~- at a coefficient of 0.813.Constructed radar chart was used to evaluate average area,average circumference and coordination of Halomonas sp.BDG-3 at different time points.Average area of cells was at a peak value of 1.39 at 3 h.Average area subsequently decreased rapidly,with a minimum of 0.003 1 at 6 h.Evaluation function f(S_i,V_i) showed that when fluoranthene was added at 3 h,Halomonas sp.BDG-3 showed the highest growth.Therefore by combining Python and Image J,a complete and intuitive method was established for image processing and evaluation of microbial Nano SIMS.This work provides a new and reliable analytical method to study degradation and metabolism of more complex environmental pollutions in microorganisms.
Nano-secondary ion technique(Nano SIMS)was used to investigate fluoranthene metabolism in Halomonassp.BDG-3.Mass signals at 12,13,16,and 26 were collected and analyzed with Image J and Python.~(12)C~- and ~(13)C~- were found to be positively correlated,with a strong correlation coefficient of 0.998.16 O-was found to be positively correlated with ~(26)(~(12)C~(14)N)~- at a coefficient of 0.813.Constructed radar chart was used to evaluate average area,average circumference and coordination of Halomonas sp.BDG-3 at different time points.Average area of cells was at a peak value of 1.39 at 3 h.Average area subsequently decreased rapidly,with a minimum of 0.003 1 at 6 h.Evaluation function f(S_i,V_i) showed that when fluoranthene was added at 3 h,Halomonas sp.BDG-3 showed the highest growth.Therefore by combining Python and Image J,a complete and intuitive method was established for image processing and evaluation of microbial Nano SIMS.This work provides a new and reliable analytical method to study degradation and metabolism of more complex environmental pollutions in microorganisms.
引文
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[2]HUA F,WANG H.Uptake modes of octadecane by Pseudomonas sp.DG17and synthesis of biosurfactant[J].J Appl Microbiol,2012,112(1):25
[3]BAUTISTA L F,SANZ R,MOLINA M C,et al.Effect of different non-ionic surfactants on the biodegradation of PAHs by diverse aerobic bacteria[J].Int Biodeterior Biodegrad,2009,63(7):913
[4]苏梦缘,李艺,王红旗,等.稳定同位素标记结合Nano SIMS亚细胞成像技术分析化合物在微生物体内代谢活性研究[J].北京师范大学学报(自然科学版),2016,52(2):223
[5]LI Y,WANG H Q,HUA F,et al.Trans-membrane transport of fluoranthene by Rhodococcus sp.BAP-1and optimization of uptake process[J].Bioresour Technol,2014,155:213
[6]WINOGRAD N.Imaging mass spectrometry on the nanoscale with cluster ion beams[J].Anal Chem,2015,87(1):328
[7]MUSAT N,HALM H,WINTERHOLLER B,et al.Asingle-cell view on the ecophysiology of anaerobic phototrophic bacteria[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(46):17861
[8]PERFUMO A,ELSAESSER A,LITTMANN S,et al.Epifluorescence,SEM,TEM and Nano SIMS image analysis of the cold phenotype of Clostridium psychrophilum at subzero temperatures[J].Fems Microbiology Ecology,2014,90(3):869
[9]李俊华.基于Python的数据分析[J].电子技术与软件工程,2018(17):167
[10]李艺,王红旗,吴枭雄,等.红球菌BAP-1对荧蒽的跨膜运输过程[J].中国环境科学,2018,38(4):1441
[11]JAIN A K,MURTY M N,FLYNN P J.Data clustering:a review[J].ACM Comput Surv,1999,31(3):264
[12]MOHEBI A,AGHABOZORGI S,TEH YING W,et al.Iterative big data clustering algorithms:a review[J].Softw-Pract Exp,2016,46(1):107
[13]索晓波,门宝辉.变异系数权重TOPSIS法在水资源综合评价中的应用[J].南水北调与水利科技,2007(5):45
[14]张良均.Python数据分析与挖掘实战[M].北京:机械工业出版社,2016
[15]方长云,胡贤巧,邵雅芳,等.基于雷达图分析法初步评价稻米食味品质的研究[J].中国稻米,2017,23(2):13
[16]魏春阳,王信民,蔡宪杰,等.基于雷达图的烤烟外观质量综合评价[J].烟草科技,2008(12):57