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医学科研型研究生常用实验技术与方法
$49.95
本着常用、基本的编写原则,让医学研究生掌握科研实验的基本功,顺利完成学业
本书是在充分了解医学科研型研究生的实验需求基础上,由科研技术人...
本书是在充分了解医学科研型研究生的实验需求基础上,由科研技术人员发挥专业特长所编写。分为三大部分,共22章。部分为实验室通识,包括现代生物医学基础研究的模式和方法、实验室生物安全、科研实验记录书写要求与规范和实验用水、溶液与缓冲液;第二部分为常用实验技术,包括细胞培养常用实验技术、常用实验动物基本知识与操作、蛋白质含量测定和酶联免疫吸附试验、PCR的原理及应用、蛋白质免疫印迹技术原理与操作、免疫细胞与组织化学技术、流式细胞术、分子克隆技术、基因修饰小鼠技术原理、中药有效成分提取分离及干燥技术等;第三部分为专题研究方法,包括细胞凋亡检测常用方法、细胞自噬检测方法、外泌体研究方法、骨代谢生物学研究方法、高效液相色谱在中药研究中的应用、中药网络药理学及常用数据库简介、组学研究方法、动物与细胞实验的系统评价与Meta分析等。
戴薇薇,博士,中西医结合生物化学专业,上海中医药大学副教授,龙华医院科技中心实验室主任,从事实验室管理与研究生科研实验教学工作,中医药延缓衰老与防治老年病机制研究。
部分·实验室通识001
章·现代生物医学基础研究的模式和方法002一·现代生物医学研究的模式002二·生物医学基础研究的三种实验系统003三·围绕“分子生物学中心法则”展开的实验方法体系004
第二章·实验室生物安全006一·实验室生物安全概念006二·实验室安全制度管理007三·实验室化学废弃物的处理010四·实验室意外事故应急预案012
第三章·科研实验记录书写要求与规范017一·实验记录的意义017二·实验记录的原则017三·实验记录书写规范要求019
第四章·实验室用水,溶液与缓冲液020一·实验室用水020二·溶液022三·溶液的浓度023四·缓冲溶液026五·常用缓冲溶液的配制029六·中药复方和中药单体浓度的计算033
第二部分·常用实验技术035
第五章·细胞培养常用实验技术036一·原代细胞分离与培养036二·细胞传代039三·细胞冻存及复苏042四·细胞计数045五·细胞活力检测047六·细胞转染050七·常用细胞培养基比较052
第六章·常用实验动物基本知识与操作053一·实验动物概念053二·实验动物分类053三·常规实验动物设施分类和使用056四·常用实验动物生物学特征057五·实验动物福利059六·实验动物基本操作060七·常用实验动物麻醉操作067八·实验动物血液标本采集070九·实验动物其他标本采集073十·实验动物的尸体剖检和脏器采出075
第七章·蛋白质含量测定和酶联免疫吸附试验077一·蛋白质含量测定077二·免疫标记技术与酶联免疫吸附试验079
第八章·PCR的原理及应用083一·PCR技术083二·实时荧光定量PCR的原理及应用091三·miRNA的PCR检测100
第九章·蛋白质免疫印迹技术原理与操作103一·蛋白质免疫印迹技术原理103二·蛋白质免疫印迹具体操作步骤110三·内参抗体的选择114四·蛋白质免疫印迹实验中的注意事项与常见问题分析114
第十章·免疫细胞与组织化学技术117一·样本组织制备117二·组织脱钙119三·组织切片制作与染色技术120四·特殊染色技术123五·免疫组织化学技术127六·免疫组织化学染色中常见的问题及对策133〖KG18mm〗〖CSX%0,0,0,30〗〖CD130mm〗〖CSX〗
第十一章·流式细胞术原理、操作与实验中的应用135一·流式细胞术概述135二·流式细胞术的原理136三·流式细胞术的基本操作143四·流式细胞术的应用152
第十二章·分子克隆原理与技术操作160一·分子克隆的基本工具160二·目标DNA序列的PCR法克隆和酶切法亚克隆164三·质粒和PCR产物的酶切及琼脂糖凝胶电泳167四·质粒酶切或PCR产物酶切后的纯化和割胶回收169五·T4 DNA连接酶介导的质粒和PCR产物的连接反应171六·质粒的转化、扩增、抽提与鉴定173
第十三章·基因修饰小鼠技术原理177一·基因编辑概述177二·Cre/LoxP系统178三·TALEN系统181四·CRISPR/Cas9基因编辑系统184
第十四章·中药有效成分提取分离及干燥技术187一·中药成分的提取方法187二·中药有效成分的分离方法189三·中药复方有效成分的提取分离191四·中药提取液的浓缩方法192五·中药提取液干燥方法及冷冻干燥技术193
第三部分·专题研究方法197
第十五章·细胞凋亡检测常用方法198一·细胞凋亡的概念198二·常用的检测方法198
第十六章·细胞自噬检测方法206一·细胞自噬概述206二·自噬的检测方法208三·细胞自噬检测方法小结217
第十七章·外泌体研究方法219一·外泌体概述219二·外泌体的常用提取方法220三·外泌体的鉴定225四·外泌体的研究策略227
第十八章·骨代谢生物学研究方法230一·骨代谢概述230二·骨组织细胞学和分子生物学230三·骨代谢细胞生物学研究方法235四·骨代谢组织生物学研究方法238
第十九章·高效液相色谱在中药研究中的应用242一·高效液相色谱简介242二·高效液相色谱仪介绍及使用方法244三·高效液相色谱法测定中药成分方法的建立246
第二十章·中药网络药理学及常用数据库简介248一·中药网络药理学及其研究思路248二·中药网络药理学常用数据库简介251
第二十一章·蛋白质组学、代谢组学技术研究方法266一·蛋白质组学及其应用266二·代谢组学及其应用272
第二十二章·动物与细胞实验的系统评价与Meta分析279一·动物与细胞实验系统评价的现状与意义279二·动物与细胞实验系统评价的基本过程280三·动物与细胞实验的检索策略的制定283四·动物和细胞实验的偏倚风险评估287五·数据分析291
联合国教科文组织将医学科研活动分为基础研究、应用研究、试验发展三大类,三者之间的非线性互动和交融关系已愈来愈引起重视,并涉及理念的转变,如ABC原则(基础研究与应用研究结合)和SED原则(科学、工程与设计融合)等。无论是基础医学还是临床医学,当前各学科的研究都体现出交叉与整合局面。现代生物医学技术发展日新月异,基础知识与相关研究不断拓宽。但医学科研课题的研究有其原则与规律,目前各类实验方法仍基本围绕“DNARNA蛋白质”中心法则这条线上的三个点,以体内实验、离体实验、体外实验的实验系统,运用细胞生物学、实验动物学、生理生化与分子生物学等技术为基础而拓展。医学科研型研究生需要接受基础技术的系统培训,才能胜任今后的科研工作。有鉴于此,在充分了解学生的实验需求基础上,科研技术人员发挥专业特长,编写了《医学科研型研究生常用实验技术与方法》。本书分为实验室通识、常用实验技术和专题研究方法三大部分,共22章,可供需要开展科研实验的医学硕士生、博士生和从事基础生物医学研究的教师、医师,以及学有所长的本科生学习。本书以“常用、基本”为原则,所述常用技术相当于科研实验的基本功,对于医学研究生而言,掌握这些常用技术可基本够用;学会这些技术,大部分医学科研实验能顺利完成。但还有许多技术,如电生理技术,未收入本书中,实验者在应用前需要有针对性地学习。限于篇幅,本书涉及的实验原理描述尽量简明扼要,而力求实用可操作性;也主要叙述生物医学基础研究,不涉及人体受试者研究。在编写过程中,由章喻、魏元基、李峻昊、张孙正远进行了校对工作,在此表示衷心感谢。由于生物医学许多概念与内容在不断地更新,编者深感自己知识与能力有限,很多方面有不足与欠缺,甚至有错漏之处,敬请专家与读者指正。
编者2021年5月
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