草地早熟禾胚胎结构石蜡切片制作方法初探
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摘要
为了掌握草地早熟禾(Poa pratensis)胚胎结构,本试验以草地早熟禾胚胎为材料,探讨了其石蜡切片制作过程中各项技术环节的不足和改良措施,分析比较了制作石蜡切片过程中在番红-固绿和埃利希氏(Ehrlich’s)苏木精-伊红两种双重染色剂及不同染色时间组合条件下所产生的效果。具体改良措施:在常规的石蜡脱水程序上将95%的酒精换为95%的伊红溶液,在透明过程中采用多梯度透明法,在浸蜡过程中采用逐级浸蜡逐级加温,在展片过程中采用40℃的温水展片捞片。结果表明,在这些改良措施的基础上,染色过程中使用埃利希氏(Ehrlich’s)苏木精-伊红双重染色30min、自来水返蓝30min,可获得染色效果较好的石蜡切片。
A study was conducted to understand embryo structure of Poa pratensis with paraffin section. The effects of Sarranine-Fast green and Ehrlich's hematoxylin-eosin double stains with different dyeing time were analyses to improve the quality of paraffin section.This study selected better preparation procedure is as following:95%eosin alcohol was selected instead of 95%alcohol for dehydration;multisegment dimethybrnzene was used instead of ethyl alcohol for transparent;rising temperature along with more wax gradually for wax dipping;slices were flatted and taken out in 40℃ warm water bath and other improvements during procedure.Based on these improvements,better quality paraffin sections were achieved with dyeing with Ehrlich's hematoxylin-eosin double stains for 30 min and returning blue with water for 30 min.
A study was conducted to understand embryo structure of Poa pratensis with paraffin section. The effects of Sarranine-Fast green and Ehrlich's hematoxylin-eosin double stains with different dyeing time were analyses to improve the quality of paraffin section.This study selected better preparation procedure is as following:95%eosin alcohol was selected instead of 95%alcohol for dehydration;multisegment dimethybrnzene was used instead of ethyl alcohol for transparent;rising temperature along with more wax gradually for wax dipping;slices were flatted and taken out in 40℃ warm water bath and other improvements during procedure.Based on these improvements,better quality paraffin sections were achieved with dyeing with Ehrlich's hematoxylin-eosin double stains for 30 min and returning blue with water for 30 min.
引文
[1]王灶安.植物显微技术[M].北京:农业出版社,1992:8.
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[7]于小刚,张文忠,韩亚东,等.粳稻颖果石蜡切片中染色时间的摸索及其解剖结构的观察[J].作物杂志,2010(5):80-83.
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[11]Zhang B,Wang Z J,Jin S H,et al.A pattern of unique embryogenesis occurring via apomixis in Carya cathayensis[J].Biologia Plantarum,2012,56(4):620-627.
[12]Wu S B,Han X M,Shang Y J,et al.Morphological characteristics of apomictic embryo in sorghum[J].Acta Agriculture Boreali-sinica,1994,9:12-15.
[13]吴树彪,尚勇进,韩雪梅,等.高粱SSA-1无融合生殖胚胎学研究[J].植物学报,1994,36(11):833-837.
[14]姚家琳,蔡得田,马平福,等.水稻无孢子生殖的胚胎学研究[J].中国水稻科学,1997,11(2):113-117.
[15]姚家琳,蔡得田.水稻HDAR细胞胚胎学研究——不定胚的发生和发育[J].武汉植物学研究,1997,15(1):5-9.
[16]杨虎彪,李晓霞,罗丽娟.植物石蜡制片中透明和脱蜡技术的改良[J].植物学报,2009,44(2):230-235.
[17]郑兴峰.植物石蜡切片法中细小或薄片状材料的包埋[J].生物学杂志,2003(4):41-43.
[18]路健,王伶.石蜡切片制作中的问题与解决[J].基础医学教育,2001(3):263-264.
[2]李正理.植物制片技术[M].第二版.北京:科学出版社,1987:138-148.
[3]Grazi F,Umaerus M,Akerberg E.Oberservations on the mode of reproduction and the embryology of Poa pratensis[J].Hereditas,1961,47:489-541.
[4]苏印泉,彭锋,黎斌,等.桦树松萝的石蜡切片方法改良及形态学研究[J].西北植物学报,2007,27(5):0859-0863.
[5]何承坤,部伸坤,李家慎.植物石蜡切片一步双重整体染色[J].福建农学院学报,1989,18(4):565-569.
[6]韩宝,冯永庆,秦岭.板栗雄花序石蜡切片改进方法的研究[J].安徽农学通报,2008,14(3):29-30.
[7]于小刚,张文忠,韩亚东,等.粳稻颖果石蜡切片中染色时间的摸索及其解剖结构的观察[J].作物杂志,2010(5):80-83.
[8]赵桂琴,曹致中.草地早熟禾无融合生殖的细胞学鉴定[J].草业学报,1997:64-70.
[9]杨宏远.用整体染色与透明技术观察胚囊、胚、胚乳和胚状体[J].植物学报,1986,28(6):575-581.
[10]李和平,孙蒙祥,蔡得田,等.草地早熟禾无融合生殖现象的研究[J].武汉植物学研究,1991(1):11-15.
[11]Zhang B,Wang Z J,Jin S H,et al.A pattern of unique embryogenesis occurring via apomixis in Carya cathayensis[J].Biologia Plantarum,2012,56(4):620-627.
[12]Wu S B,Han X M,Shang Y J,et al.Morphological characteristics of apomictic embryo in sorghum[J].Acta Agriculture Boreali-sinica,1994,9:12-15.
[13]吴树彪,尚勇进,韩雪梅,等.高粱SSA-1无融合生殖胚胎学研究[J].植物学报,1994,36(11):833-837.
[14]姚家琳,蔡得田,马平福,等.水稻无孢子生殖的胚胎学研究[J].中国水稻科学,1997,11(2):113-117.
[15]姚家琳,蔡得田.水稻HDAR细胞胚胎学研究——不定胚的发生和发育[J].武汉植物学研究,1997,15(1):5-9.
[16]杨虎彪,李晓霞,罗丽娟.植物石蜡制片中透明和脱蜡技术的改良[J].植物学报,2009,44(2):230-235.
[17]郑兴峰.植物石蜡切片法中细小或薄片状材料的包埋[J].生物学杂志,2003(4):41-43.
[18]路健,王伶.石蜡切片制作中的问题与解决[J].基础医学教育,2001(3):263-264.