三种不同固定液对豚鼠眼球的固定效果比较
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摘要
目的:用过碘酸雪夫氏(Periodic Acid-Schiff,PAS)染色法比较4%多聚甲醛固定液、4%戊二醛固定液和Davidson固定液固定豚鼠眼球的固定效果,筛选最佳眼球固定液和固定时间。方法:取正常豚鼠眼球分6组,每组5只,Ⅰ组眼球放入4%多聚甲醛固定液固定24h、Ⅱ组眼球放入4%戊二醛固定液固定24h;Ⅲ～Ⅴ组眼球放入Davidson固定液分别固定3、6、24h;Ⅵ组眼球在Davidson固定液中固定3h后转移到10%中性甲醛中再固定48h。常规制片、PAS染色、显微镜观察,比较不同固定方法对组织的固定效果。结果:Davidson固定液固定3h的固定效果最为理想,Davidson固定液固定3h后转移到10%中性甲醛再固定48h的固定效果与Davidson固定液固定3h的固定效果接近,这两组固定液固定的眼球切片均结构完整、层次清晰,视网膜各层细胞排列整齐。结论:Davidson固定液对豚鼠眼球的固定效果明显优于其他两种固定液,豚鼠眼球用Davidson固定液固定后,可将其转移到中性甲醛中长期保存,其固定效果不受影响。
AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4% paraformaldehyde solution, 4% glutaraldehyde solution and Davidson solution.METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group I-II were fixed in 4%paraformaldehyde solution and 4% glutaraldehyde solution for 24 h,respectively; Group III-V were fixed in Davidson solution for 3,6 and 24 h, respectively; and Group VI were fixed in Davidson solution for 3 h and then transferred into 10% neutral formaldehyde solution for48 h. All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.RESULTS:It was found that the group which was fixed by Davidson solution for 3 h,remained the most complete structure for PAS staining( Group III). While the effect of fixation for the group which was transferred into 10%neutral formaldehyde solution for preserving for 48 h after fixing in Davidson solution for 3 h was also acceptable for PAS staining( Group VI). The retinal cells remained clear and in order for both groups which was mentioned above.CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3 h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for3 h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.
AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4% paraformaldehyde solution, 4% glutaraldehyde solution and Davidson solution.METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group I-II were fixed in 4%paraformaldehyde solution and 4% glutaraldehyde solution for 24 h,respectively; Group III-V were fixed in Davidson solution for 3,6 and 24 h, respectively; and Group VI were fixed in Davidson solution for 3 h and then transferred into 10% neutral formaldehyde solution for48 h. All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.RESULTS:It was found that the group which was fixed by Davidson solution for 3 h,remained the most complete structure for PAS staining( Group III). While the effect of fixation for the group which was transferred into 10%neutral formaldehyde solution for preserving for 48 h after fixing in Davidson solution for 3 h was also acceptable for PAS staining( Group VI). The retinal cells remained clear and in order for both groups which was mentioned above.CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3 h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for3 h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.
引文
1李国平,吴建峰,叶翔,等.负透镜诱导型近视豚鼠视网膜中谷氨酸及其受体的表达.眼科新进展2016;36(10):910-914
2 曾官鹏,严丽英,徐双,等.MFN1在透镜诱导豚鼠近视模型视网膜上的表达及其意义.眼科新进展2016;36(1):1-3,11
3 王伯云,李玉松,黄高晟,等.病理技术.北京:人民卫生出版社2001:67-74
4 罗灿峤,莫木琼,聂钊铭,等.比较5种混合固定液对小鼠眼球标本制片质量和染色效果的影响.中国卫生检验杂志2015;25(13):2065-2067
5 陈锦,史炯,孟凡青,等.PAS改良染色法及其在病理诊断中的应用.现代肿瘤医学2015;23(21):3069-3071
6 黄宏森,张超,郭晓珍,等.不同固定液及固定时间对小鼠肝组织结构的影响.广州医学院学报2007;35(2):65-68
7 陈广斌,谢玲,林坚涛.不同固定方法对新生大鼠脑组织切片的影响.中国民康医学2008;20(15):1730-1731
8 孙自强,王军,陈明亮,等.小鼠眼球标本石蜡切片制作方法的改进.河南大学学报(医学版)2015;34(1):36-39
9 王春霞,于紫燕,张劲松.大鼠视网膜标本石蜡切片的固定方法.解剖学研究2012;34(4):316-318
10 李晶晶,朱鸿,施彩虹.三种方法对大鼠视网膜固定效果的比较研究.上海交通大学学报(医学版)2011;31(8):1105-1107
11 李天全.生物组织固定剂戊二醛的化学和应用.生物医学工程学杂志1988;5(2):118-124
12 陶亚群,李贵苹,易静,等.兼顾免疫组化和电镜样品的戊二醛-多聚甲醛混合固定液的应用.中国细胞生物学学报2016;38(12):1518-1522
13 Moore KL.The sex chromatin in benign tumours and related conditions in man.Br J Cancer 1955;9(2):246-252
14 李雷,严建燕,苏欣,等.大鼠睾丸组织固定法的优选研究.中国比较医学杂志2013;23(6):20-23,80-82
15 赵玉琼,李瑞生,陈华.三种固定液对大、小鼠肠道组织石蜡切片质量的影响.中国比较医学杂志2014;24(11):53-56
16 盛嬅,王辉,陆姮磊,等.Davidson's固定液用于睾丸和眼球固定的优势.毒理学杂志2007;4:291
17 Chidlow G,Daymon M,Wood JP.Localization of a wide-ranging panel of antigens in the rat retina by immunohistochemistry:comparison of Davidson's solution and formalin as fixatives.Histochem Cytochem2011;59(10):884-898
18 Latendresse JR,Warbrittion AR.Jonassen H,et al.Fixation of testes and eyes using a modified Davidson's fluid:comparison with Bouin's fluid and conventional Davidson fluid.Toxicol Pathol 2002;30(4):524-533
2 曾官鹏,严丽英,徐双,等.MFN1在透镜诱导豚鼠近视模型视网膜上的表达及其意义.眼科新进展2016;36(1):1-3,11
3 王伯云,李玉松,黄高晟,等.病理技术.北京:人民卫生出版社2001:67-74
4 罗灿峤,莫木琼,聂钊铭,等.比较5种混合固定液对小鼠眼球标本制片质量和染色效果的影响.中国卫生检验杂志2015;25(13):2065-2067
5 陈锦,史炯,孟凡青,等.PAS改良染色法及其在病理诊断中的应用.现代肿瘤医学2015;23(21):3069-3071
6 黄宏森,张超,郭晓珍,等.不同固定液及固定时间对小鼠肝组织结构的影响.广州医学院学报2007;35(2):65-68
7 陈广斌,谢玲,林坚涛.不同固定方法对新生大鼠脑组织切片的影响.中国民康医学2008;20(15):1730-1731
8 孙自强,王军,陈明亮,等.小鼠眼球标本石蜡切片制作方法的改进.河南大学学报(医学版)2015;34(1):36-39
9 王春霞,于紫燕,张劲松.大鼠视网膜标本石蜡切片的固定方法.解剖学研究2012;34(4):316-318
10 李晶晶,朱鸿,施彩虹.三种方法对大鼠视网膜固定效果的比较研究.上海交通大学学报(医学版)2011;31(8):1105-1107
11 李天全.生物组织固定剂戊二醛的化学和应用.生物医学工程学杂志1988;5(2):118-124
12 陶亚群,李贵苹,易静,等.兼顾免疫组化和电镜样品的戊二醛-多聚甲醛混合固定液的应用.中国细胞生物学学报2016;38(12):1518-1522
13 Moore KL.The sex chromatin in benign tumours and related conditions in man.Br J Cancer 1955;9(2):246-252
14 李雷,严建燕,苏欣,等.大鼠睾丸组织固定法的优选研究.中国比较医学杂志2013;23(6):20-23,80-82
15 赵玉琼,李瑞生,陈华.三种固定液对大、小鼠肠道组织石蜡切片质量的影响.中国比较医学杂志2014;24(11):53-56
16 盛嬅,王辉,陆姮磊,等.Davidson's固定液用于睾丸和眼球固定的优势.毒理学杂志2007;4:291
17 Chidlow G,Daymon M,Wood JP.Localization of a wide-ranging panel of antigens in the rat retina by immunohistochemistry:comparison of Davidson's solution and formalin as fixatives.Histochem Cytochem2011;59(10):884-898
18 Latendresse JR,Warbrittion AR.Jonassen H,et al.Fixation of testes and eyes using a modified Davidson's fluid:comparison with Bouin's fluid and conventional Davidson fluid.Toxicol Pathol 2002;30(4):524-533