[Objective] The research aimed to study a paraffin method without using the microtome,and also introduced an analysis method for optical information of the plant anatomical digital photographs.[Method] The plant mater...[Objective] The research aimed to study a paraffin method without using the microtome,and also introduced an analysis method for optical information of the plant anatomical digital photographs.[Method] The plant material softened or not was embedded in paraffin according to the paraffin method.Cut the thin paraffin sections from the paraffin block with a sharp two-sided blade under anatomical lens.The thin material sections rolled up when they were cut off.Took the section rolls to a slide,and then heated them to melt the paraffin section roll.When the paraffin melted,the sections of plant material were rolled out.After the common or simplified procedures of staining and mounting,the preparations were finished.When an anatomical digital photograph was processed,copy it into the word file and two copies of the original photograph were obtained.One copy was selected to make it to be a negative photograph,and then press the key ＂Press Screen＂ to copy the screen frame.After it was copied into the word file,cut of the unnecessary parts and other operations were carried out,then processed photograph was obtained.[Results] The anatomical preparation for research was gotten.The analyzed digital photograph of the leaf structure of Salix matsudana var.matsudana f.tortuosa has some a three-dimensional effect,and the different leaf structures and cells,e.g.cuticle,cell wall,protoplast,vein,etc.can be identified easily.[Conclusion]The paraffin method without using the microtome has advantages of low cost and higher efficiency,which could be applied by the beginner or in the time without a microtome to be used.The analysis of the plant anatomical digital photographs can acquire more structural information than the original digital photographs,which shows the potentiality and prospects of the optical information analysis of the microscopic imagery and its digital photograph.展开更多
本研究以10例甲醛固定石蜡包埋组织(FFPET)作为疑难检材,将其制成蜡膜,分别以二甲苯-Chelex100法、脱蜡液-Chelex100法、脱蜡液-Vazyme FastPure FFPE DNA Isolation试剂盒(简称FP试剂盒)、脱蜡液-北京天根石蜡包埋组织基因组提取试剂盒...本研究以10例甲醛固定石蜡包埋组织(FFPET)作为疑难检材,将其制成蜡膜,分别以二甲苯-Chelex100法、脱蜡液-Chelex100法、脱蜡液-Vazyme FastPure FFPE DNA Isolation试剂盒(简称FP试剂盒)、脱蜡液-北京天根石蜡包埋组织基因组提取试剂盒法(简称天根试剂盒)提取DNA,进行紫外分光光度计测定(浓度、纯度)及短串联重复序列(STR)分型检测(23个常染色体),通过比较分析FFPET在4种不同提取方法中的提取质量、STR分型质量和检出率,从而寻求一种高效简便、经济实用、方便快捷的甲醛固定石蜡包埋组织中DNA提取方法。结果表明:应用4种方法提取的石蜡包埋组织蜡膜DNA经紫外分光光度计测定后显示,应用2种Chelex100法提取的DNA浓度高于2款试剂盒,试剂盒提取的DNA的纯度优于另外2种方法,以上差异均具有统计学意义(P<0.05);FP试剂盒提取DNA的平均浓度和纯度均优于天根试剂盒,无显著性差异(P>0.05);STR分型普遍存在不同基因座和同一基因座不同等位基因之间的扩增不均衡,呈现前高后低状态;STR平均检出率为FP试剂盒>天根试剂盒>Chelex100法,2种Chelex100法STR基因座检出率比较无显著差异(P>0.05),但分别和另外2种方法试剂盒方法相比有显著差异(P<0.05),FP试剂盒和天根试剂盒法相比有显著差异(P<0.05)。因此,在涉及甲醛固定石蜡包埋组织这类疑难检材的法医DNA鉴定案件时,脱蜡液-Vazyme FastPure FFPE DNA Isolation试剂盒法优于其余3种提取DNA方法。展开更多
基金Supported by National Natural Science Foundation of China(30770124)~~
文摘[Objective] The research aimed to study a paraffin method without using the microtome,and also introduced an analysis method for optical information of the plant anatomical digital photographs.[Method] The plant material softened or not was embedded in paraffin according to the paraffin method.Cut the thin paraffin sections from the paraffin block with a sharp two-sided blade under anatomical lens.The thin material sections rolled up when they were cut off.Took the section rolls to a slide,and then heated them to melt the paraffin section roll.When the paraffin melted,the sections of plant material were rolled out.After the common or simplified procedures of staining and mounting,the preparations were finished.When an anatomical digital photograph was processed,copy it into the word file and two copies of the original photograph were obtained.One copy was selected to make it to be a negative photograph,and then press the key ＂Press Screen＂ to copy the screen frame.After it was copied into the word file,cut of the unnecessary parts and other operations were carried out,then processed photograph was obtained.[Results] The anatomical preparation for research was gotten.The analyzed digital photograph of the leaf structure of Salix matsudana var.matsudana f.tortuosa has some a three-dimensional effect,and the different leaf structures and cells,e.g.cuticle,cell wall,protoplast,vein,etc.can be identified easily.[Conclusion]The paraffin method without using the microtome has advantages of low cost and higher efficiency,which could be applied by the beginner or in the time without a microtome to be used.The analysis of the plant anatomical digital photographs can acquire more structural information than the original digital photographs,which shows the potentiality and prospects of the optical information analysis of the microscopic imagery and its digital photograph.
文摘本研究以10例甲醛固定石蜡包埋组织(FFPET)作为疑难检材,将其制成蜡膜,分别以二甲苯-Chelex100法、脱蜡液-Chelex100法、脱蜡液-Vazyme FastPure FFPE DNA Isolation试剂盒(简称FP试剂盒)、脱蜡液-北京天根石蜡包埋组织基因组提取试剂盒法(简称天根试剂盒)提取DNA,进行紫外分光光度计测定(浓度、纯度)及短串联重复序列(STR)分型检测(23个常染色体),通过比较分析FFPET在4种不同提取方法中的提取质量、STR分型质量和检出率,从而寻求一种高效简便、经济实用、方便快捷的甲醛固定石蜡包埋组织中DNA提取方法。结果表明:应用4种方法提取的石蜡包埋组织蜡膜DNA经紫外分光光度计测定后显示,应用2种Chelex100法提取的DNA浓度高于2款试剂盒,试剂盒提取的DNA的纯度优于另外2种方法,以上差异均具有统计学意义(P<0.05);FP试剂盒提取DNA的平均浓度和纯度均优于天根试剂盒,无显著性差异(P>0.05);STR分型普遍存在不同基因座和同一基因座不同等位基因之间的扩增不均衡,呈现前高后低状态;STR平均检出率为FP试剂盒>天根试剂盒>Chelex100法,2种Chelex100法STR基因座检出率比较无显著差异(P>0.05),但分别和另外2种方法试剂盒方法相比有显著差异(P<0.05),FP试剂盒和天根试剂盒法相比有显著差异(P<0.05)。因此,在涉及甲醛固定石蜡包埋组织这类疑难检材的法医DNA鉴定案件时,脱蜡液-Vazyme FastPure FFPE DNA Isolation试剂盒法优于其余3种提取DNA方法。
