安徽医科大学：《医学细胞生物学》课程教学资源（实践指导，打印版）细胞器的分级分离

安徽医科大学生命科学学院 School Of Life Sciences Aukui Wedieal Hniversity 细胞生物学实验之 细胞器的分级分离 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.初步掌握细胞器的分离原理及方法。 2.学习并掌握高速离心机和匀浆器的使 用方法。 3.掌握差速离心法和密度梯度离心法的 原理。 2 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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Cell Membrane Centrosome 细 Lysosome】 别 Cyteplasm [实验原理] Nucleus Rough ER Smeoth ER Nuclear 1.细胞器的分离 Membrane 制备细 分级 组分分 胞匀浆 分离 析鉴定 ● 机械法 ● 差速离心法 ●形态鉴定 ●物理法 ● 密度梯度离心法 ●功能鉴定 ● 化学法 3 鱼 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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[实验原理] 2.分级分离方法 差速离心法(Differential centrifugation) 速度区带离心法 (Rate-zone 心方 密度梯度离心法 centrifugation) (Density gradient centrifugation) 等密度区带离心法 法 (Isopycnic density gradie nt zonal centrifugation) 超速离心法(Jltracentrifugation) 静 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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[实验原理] 2.分级分离方法 (1)差速离心法(Differential centrifugation) SUPERNATANT 1 SUPERNATANT 2 SUPERNATANT 3 LOW-SPEED MEDIUM-SPEED HIGH-SPEED VERY HIGH-SPEED CENTRIFUGATION CENTRIFUGATION CENTRIFUGATION CENTRIFUGATION 的 cell homogenate PELLET 1 PELLET 2 PELLET 3 PELLET 4 whole cells mitochondria microsomes ribosomes nuclei lysosomes other small vesicies viruses cytoskeletons peroxisomes large macromolecules 完整细胞、细胞核、 线粒体、溶酶 微体、囊泡等 核糖体、病毒等 细胞骨架 体、过氧化物 酶体 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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[实验原理] 2.分级分离方法 (2)速度区带离心法(Rate-zone centrifugation) 预先装入有一定密度梯度的材料（蔗糖或甘油)， 利用各种颗粒在梯度液中的沉降速度不同，使具有相同 沉降速度的颗粒处于同一梯度层内。 样品 离心 分部收集 浅梯度 注：待分离颗粒密度 比介质密度都要大： 不能离心太久， 否则两种颗粒都会沉 1098765432 到底部。 鱼钢 This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more informatior

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低速离心机≤5000r/min 68● 90 10000r/min30000r/min This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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[实验原理] 3.离心机的使用 ·离心机在运转时，不得移动离心机。 离心机必须放置在坚固水平的台面上，并且保持离心 机的平稳，以免产生振动。 放置离心管时需注意配平：加液应称量平衡；试管必 须对称放入 加液体积应小于离心管体积2/3 若运行时有离心试管破裂，会引起较大振动应立即停 机处理 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 大鼠肝细胞细胞核及线粒体的分离 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： 大鼠肝脏 器具： 制备匀浆：玻璃匀浆器、滤网、烧杯 分离：低速离心机、高速离心机、1.5mLEp管、5mL离心管 形态鉴定：普通光学显微镜、载玻片、盖玻片、染缸 试剂： 。 生理盐水、PBS、0.25mol/L的蔗糖溶液、0.34mol/L蔗糖-0.5 mmol/LMg(Ac2溶液、0.88mol/L蔗糖-0.5mmol/LMg(Ac2溶液、95% 淳、 甲基绿-派洛宁染液、丙酮、0.2%的詹纳斯绿B。(mol/儿=M This document is produced by trial version of Print2Flash.Visit www.print2flash.com for more information

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