Methyl Green-Pyronin Staining Solution

甲基绿-派洛宁染色液是一种组织或细胞染色时常用的可以把细胞核染成绿色或蓝绿色，把细胞浆和细胞核中的核仁染成红色的染色液。甲基绿可以和细胞核中的DNA结合，从而产生细胞核染色；而派洛宁可以和细胞浆或核仁中的RNA结合，从而可以使细胞浆和核仁被染色。 操作流程： 1 样品处理 1.1 对于石蜡切片：二甲苯中脱蜡 2×5min；无水乙醇 5 min、90%乙醇 2 min、70%乙醇 2 min；PBS 漂洗2 min。 1.2 对于冰冻切片：PBS 漂洗 2 min。 1.3 对于培养细胞：用 4%多聚甲醛固定 10 min 以上，PBS 洗涤 2×2 min。 2 甲基绿-派洛宁染色对于上述处理好的样品： 2.1 甲基绿-派洛宁染色液染色 5 ~10min 左右(可以根据染色结果和要求调整时间)。2.2 蒸馏水洗涤 2-3 次(每次数秒钟)。 3 脱水、透明、封片或进行其它染色 3.1 丙酮脱水 3 次，每次 1~2sec(3 dips)。 3.2 换用丙酮/二甲苯(1:1)洗涤 2 次，每次 1~2min。 二甲苯透明 2×1~2min。3.3 用中性树胶或其它封片剂封片。 3.4 显微镜下观察，细胞核呈绿色或蓝绿色，而细胞浆和核仁呈红色。 染色结果：细胞核染成绿色或蓝绿色，细胞浆和细胞核中的核仁染成红色。 注意事项： 1 需自备 4%多聚甲醛、梯度乙醇、二甲苯，中性树胶或其它封片剂。2 第一次使用本试剂盒时建议先取 1~2 个样品做预实验。 温馨提示： 为了您的自身安全，使用试剂前，请做好防护，如穿实验服，带手套等。 Methylgreen-pyronine is a common dye used in tissue or cell staining to color the nucleus green or blue-green and the nucleoli in the cytoplasm and nucleus red. Methyl green can bind to DNA in the nucleus, thus producing nuclear staining; Pyronin can bind to RNA in the cytoplasm or nucleolus, so that the cytoplasm and nucleolus can be stained. Operation process: 1 Sample handling 1.1 For paraffin sections: dewaxing in xylene for 2×5min; Anhydrous ethanol 5 min, 90% ethanol 2 min, 70% ethanol 2 min; Rinse with PBS for 2 min. 1.2 For frozen sections: Rinse with PBS for 2 min. 1.3 For cultured cells: fixed with 4% paraformaldehyde for more than 10 min and washed with PBS for 2×2 min. 2 methyl green-Pyronine staining for the above treated samples: 2.1 Methyl green-Pyronine dyeing solution staining for about 5 ~10min (the time can be adjusted according to the dyeing results and requirements). 2.2 Wash distilled water 2-3 times (each time for seconds). 3 Dehydration, transparency, sealing or other dyeing 3.1 Dehydration of acetone 3 times, 1~2sec(3 dips) each time. 3.2 Wash with acetone/xylene (1:1) twice, 1~2min each time. Xylene transparent 2×1~2min. 3.3 Seal the tablet with neutral gum or other sealant. 3.4 Microscopically, the nucleus appears green or blue-green, while the cytoplasm and nucleolus appear red. Staining results: the nucleus is dyed green or blue-green, and the nucleoli in the cytoplasm and nucleus are dyed red. Note: 1 Need to bring 4% polyformaldehyde, gradient ethanol, xylene, neutral gum or other tablets. 2 When using this kit for the first time, it is recommended to take 1 to 2 samples for pre-test. Warm reminder: For your own safety, please take good protection before using the reagent, such as wearing a lab coat, gloves, etc.

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