摘要
目的:Nd:YAP激光辐照对雨生红球藻生理效应的荧光分析。方法:利用Nd:YAG激光(功率10W,辐照剂量为15s、35s、55s)辐照雨生红球藻,在对辐照细胞进行生长测定以及色素吸收光谱分析的基础上,进一步利用激光共聚焦扫描显微镜(LSCM),在波长488nm的Ar+激发光条件下,对细胞的自体荧光以及吖啶橙染色的细胞核荧光物质进行定位定量分析,获得细胞自发荧光光谱和细胞核荧光物质的荧光光谱。结果:①低剂量的Nd:YAG激光辐照(15s左右)对藻细胞有促长作用,生长速率提高20.3%,色素吸收峰的峰值提高25.3%。较高剂量的Nd:YAG激光辐照(35s、55s)对藻细胞生长有抑制作用,并有明显的致死、致突效应。②对自体荧光的分析结果表明,与对照组相比,低剂量激光辐照组的细胞,在荧光光谱的峰型及峰值上变化不大,在682nm处均有较强的荧光发射,而高剂量激光辐照组的细胞多无明显的荧光发射。③对细胞核荧光物质的分析,雨生红球藻细胞在530nm(DNA)和640nm(RNA)处均有荧光发射峰。与对照组相比,低剂量激光辐照组的DNA荧光发射有所增强,但RNA的荧光发射则有所减弱;高剂量激光辐照组的核物质荧光发射谱,在峰型上与对照组存在差异,荧光强度也明显下降。结论:低剂量的Nd:YAP激光辐照对细胞核的DNA合成与复制有一定的刺激作用,可促进光合色素的合成并提高其对光能的吸收效率,从而增强光合活性,促进细胞的增殖与生长;高剂量激光辐照则对细胞的DNA有损伤作用,是致死率上升并发生突变的可能原因。
Objectlve: The fluorescence spectra of physiological effect of Haematococcus was measured in this study. Method: Nd:YAP Laser (wavelength 1340nm,power10w and with irradiate does of 15 sec,35sec,55sec. )was used to irradiate the cells of Haematococcus pluxialis. Irradiated cells was studied flwough measuring cell number and the quantity of chlorophyll, then using LSCM, autofluorescenee of ceils and nueleolus fluorescence matter by AO-dyeing were studied with orientation and ration. Then attaining autofluorescenee of cells and fluorescent spectrum of nueleolus fluorescence matter by Ar^+ Confocal Laser Scanning Mieroscope(LCSM, excited wavelength 488nm). Result: Firstly, the experimental results showed that the lower irradiated dose(15sec) accelerated the growth of cells,the growth rates promoted by 20.3% ,the peak value of the absorption spectrum of chlorophyll promoted by 25.3%. The higher irradiated dese(35sec, 55see) inhibited the growth of cells, and also had distinct death and mutation. Secondly, with studying autofluorescence show that the ceils in the different physiological state have different fluorescence and intension of fluorescence. Compare to the contrast, the low dose irradiated cells have no change of peak value and peak type, and they have stronger fluorescence eradiation,but high dose have no distinct fluorescence eradiafion. Thirdly, ucleolus matter fluorescence was studied show that the ceils of Haematococcus pluxialis have fluorescent eradiation peak with wavelength 530nm(DNA) ,640nm(RNA). Compare to the contrast,the low irradiated dose advanced fluorescence eradiation of DNA,but would induce fluorescence eradiation of RNA. nucleolus fluorescence eradiation spectnnn of the high irradiated dose is different from contrast in peak state, and the intension of fluorescence had distinctly fallen. Conctusion: the lower irradiated dose(15sec) can accelerate synthe^izafion and replication of DNA in nucleolus, also can accelerate synthesization of photosynthetic pigment, enhance absorbable efficiency of lightenergy, consequently, increase the synthetical activeness, accelerate proliferation and growth of cell; the higher irradiated dose have damnification of DNA, which could increase ratio of death,and aslo can engender mutation.
出处
《生物技术》
CAS
CSCD
2008年第3期26-31,共6页
Biotechnology
基金
福建省自然科学基金项目(A0610006
2006J0021)
福建省科技计划项目(K04031)
光子技术福建省重点实验室开放课题(FP0407)