安徽医科大学生命科学学院 Sckool Aike Scicnees Anbui medial Uaiwersity 细胞生物学实验之 碱性蛋白与酸性蛋白 的显示 By生物学系-李超 2016-04 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[目的要求] 1.学会制作血涂片。 2.了解碱性蛋白与酸性蛋白的显示方法的 原理及操作步骤。 3.了解碱性蛋白与酸性蛋白的分布。 2 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验原理] 以酸性氨基酸成分为 主的蛋白质为酸性蛋白；以 碱性氨基酸成分为主的蛋白 质为碱性蛋白。细胞核内有 组蛋白（碱性蛋白）及少量 酸性蛋白，细胞质中主要有 酸性蛋白，标本经三氯醋酸 处理抽提掉核酸后，用不同 pH值的快绿染液分别染色 使细胞内的酸性蛋白和碱性 蛋白质显示出来。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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本期实验内容 蟾蜍心包血细胞中碱性蛋白的显示 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验用品] 材料：蟾蜍(or牛蛙)血涂片 器具：普通光学显微镜、恒温水浴箱、载玻片、染色 缸等。 试剂：70%乙醇溶液，5%三氯醋酸溶液，0.2%快绿染 液，0.005%Na2CO3溶液，碱性蛋白染色液（pH8.0): 0.2%快绿和等量0.005%Na2C03溶液混合而成。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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双毁髓法处死蟾蜍 幽 双毁髓法处死蛙类，即用解剖针捣毁其脑组织和脊髓 。具体方法是，左手握住蟾蜍的身体和四肢，使其腹 部贴着掌心，食指压住蟾蜍头部前端使其尽量腹屈。 在头与躯干之间可触及一凹陷（即枕骨大孔所在处）， 右手持解剖针直插入此凹陷约|~2mm,随即将针尖转 向头侧插入颅腔捣毁脑组织。然后将解剖针抽回并转 向尾侧刺入脊椎管内捣毁其脊髓。如此直至其四肢松 软，呼吸消失为止。 当 以破坏脊髓法处死蟾蜍，将其腹面向上放人蜡盘中， 剪开胸腔，打开心包。小心将心脏剪一小口，取心脏 血一滴滴在干净载玻片一端，推片，按此法制备一张 血涂片，室温晾干。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more informatior
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图16-1蛙类双毁糖法 7 鱼 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验步骤] 1.制作血涂片 30-45 剧11-1血涂片的推片姿势和方法 1.取一滴血滴于玻片的一端 2.以另一玻片以30-45度的角度匀速向前推片，中间 不要停顿，不可反复涂片，动作要快以防止血液 凝固。 3.自然干燥后固定。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[实验步骤] 2.固定 新鲜血涂片自然干燥10min,入70%乙醇溶液中固定10min。 3.三氯醋酸处理 取出晾干后，入70C的5%三氯醋酸溶液15-20min。 4.染色 从三氯醋酸中取出后，用自来水冲洗3次，每次5min。 随后 插入碱性蛋白染色液中染色45min。 5.镜检 从染液中取出后，用自来水冲洗。风干后在显微镜下观察。 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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[作业题] 1.绘出细胞中碱性蛋白的分布图。 2.细胞中碱性蛋白与酸性蛋白的分布有何区别？ 3.该实验中5%三氯醋酸的作用是什么？ This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information
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