细胞工程 实验技术课件.pptVIP

10/2005 CE Lecture 1细胞工程实验室基本实验技术 细胞培养的基本概念 细胞培养的基本条件 细胞培养的基本技术 紫外线消毒主要用于培养室空气、操作台、塑料培养　　　　　　皿和培养板等表面消毒 Pipettes, bottles, flasks, petri dishes ? 塑料 -- better attachment and cell growth of monolayer cell culture -- more expensive than reusable glass -- suitable for storage at 4oC ? 玻璃 -- Can withstand temperatures 0oC use for maintaining frozen stocks -- Steam sterilize by autoclaving * relies on steam pressure and high heat * use fast-dry cycle to dry condensate * use “wet” cycle for salt solutions to prevent evaporation * loosely place caps on bottles 实验室安全 培养基的成分及配置 细胞培养的基本操作 Filtration Sterilization of solutions for maintenance growth or treatment Do not autoclave nutrient medium—heat will destroy some of the compounds in the medium Pass through sterile filter with small pore size (22 μm) use sterile techniques 浸泡（自来水）、刷洗（洗衣粉）、酸泡（24小时） 流水冲洗、蒸溜水浸泡和冲洗、50℃烘干 The End 移液器 细胞检测设备：Inverted microscope It is vital to look at cultures regularly A morphological change is often the first sign of deterioration in culture Inverted because it is not good to open tissue culture dishes ? It should have phase contrast – Most cells are not dense enough to be visible in a regular light without staining 酶标仪 微孔板震荡器 细胞检测设备 细胞检测设备：高速离心机 细胞保存设备： Equipment for cryopreservation 液氮 Liquid nitrogen – Liquid phase (-196oC) or vapor phase (-156oC) 可调节速冻速率的容器 Fancy freezing chambers to regulate the freezing rate – Styrofoam containers work too 细胞保存设备：液氮罐 CE1-3 细胞培养的基本技术 实验室安全：制度 1、准入制度 2、安全制度 3、卫生消毒制度 4、出入制度 实验室安全： Basic safety considerations Safety applies both ways – Protect your cells and protect yourself General safety rules Aseptic vs. antiseptic technique 实验室安全： Safety issues Broken glass Sharp instruments Electricity Frost bite from liquid nitrogen handling Burns (if you use Bunsen burner) Irradiation if isotopes are used Infection Chemical toxicity Biohazards 实验室安全： Safety Disinfection Work surfaces, culture waste and equipmen