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时间:2019-10-12
《番红O-快绿染色》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、番红O-固绿染色方法1、0.02%固绿溶液:固绿0.02g,蒸馏水100ml2、0.1%番红O溶液:番红0.1g,蒸馏水100ml3、1%盐酸酒精分化液:弄HCI1份,95%乙醇99份染色1、石蜡切片脱蜡至水2、苏木精染细胞核1-3min,显微镜观察染色情况决定染色时间长短3、自来水充分洗涤,去除残留苏木精。4、1%盐酸酒精分化15-30s,分化时间视颜色深浅而定。5、自来水充分洗涤,去除残留染液。6、0.02%固绿水溶液染色3min,固绿为一种酸性染料,呈碱性细胞质或细胞外基质呈绿色7、1%冰醋酸洗涤切片,
2、去除残留固绿8、0.1%番红O染色3min,番红是一种碱性染料,呈酸性细胞质或细胞外基质呈橙黄/红色。9、95%乙醇侵洗,洗去残留番红10、95%乙醇、无水乙醇脱水、二甲苯透明,中性树胶封片。番红O-固绿染色改良版1、石蜡切片脱蜡至水2、Weigert氏苏木素染色3分钟3、1%盐酸酒精分化4、水洗5、0.2%固绿水溶液内染色5分钟6、水洗7、0.1%番红O水溶液内染色1-2分钟8、水洗9、0.1%醋酸水溶液分化10、水洗11、95%乙醇、无水乙醇脱水、二甲苯透明,中性树胶封片。SolutionsandReag
3、ents: Weigert'sIronHematoxylinSolution: StockSolutionA: Hematoxylin-----------------------------1g 95%Alcohol-----------------------------100ml StockSolutionB: 29%Ferricchlorideinwater-----------4ml Distilledwater------------
4、--------------95ml Hydrochloricacid,concentrated----1ml Weigert'sIronHematoxylinWorkingSolution: MixequalpartsofstocksolutionAandB.Thisworkingsolutionisstableforabout4weeks. 0.05%FastGreen(FCF)Solution: Fastgreen,FCF,C.I.42053---------
5、----0.5g Distilledwater----------------------------1000ml 1%AceticAcidSolution: Aceticacid,glacial-------------------------1ml Distilledwater------------------------------99ml 0.1%SafraninOSolution: SafraninO,C.I.50240-------------------
6、-0.1g Distilledwater-----------------------------100ml Procedure: 1. Deparaffinizeandhydrateslidestodistilledwater.2. StainwithWeigert’sironhematoxylinworkingsolutionfor10minutes.3. Washinrunningtapwaterfor10minutes.4. Stainwithfastgreen(FCF)so
7、lutionfor5minutes.5. Rinsequicklywith1%aceticacidsolutionfornomorethan10–15seconds.6. Stainin0.1%safraninOsolutionfor5minutes.7. Dehydrateandclearwith95%ethylalcohol,absoluteethylalcohol,andxylene,using2changeseach,2minuteseach.8. Mountusingresinous
8、medium.
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