摘要:果蝇Schneider 2 (S2) 细胞培养简单,是生化和免疫组化等实验中最常用的果蝇细胞系。该细胞系于1972年由Imogene Schneider从果蝇胚胎中分离得到 (Schneider, 1972),是一种血淋巴来源的细胞。本方案主要介绍果蝇S2细胞的培养、复苏和冻存操作。
材料与试剂
- 细胞冻存管 (Corning, catalog number: 430659)
- 10 ml塑料移液管 (Corning, Costar, catalog number: 4488)
- 15 ml塑料离心管 (Thermo Fisher Scientific, Nunc, catalog number: 339650)
- 50 ml塑料离心管 (Thermo Fisher Scientific, Nunc, catalog number: 339652)
- 6孔细胞培养板 (Thermo Fisher Scientific, Nunc, catalog number: 140675)
- 24孔细胞培养板 (Thermo Fisher Scientific, Nunc, catalog number: 142475)
- 细胞培养瓶 (Thermo Fisher Scientific, Nunc, catalog number: 156367)
- 细胞培养皿 (Thermo Fisher Scientific, Nunc, catalog number: 150464)
- 一次性无菌过滤装置 (Thermo Fisher Scientific, Nalgene, catalog number: 1660045)
- 程序降温盒 (Thermo Fisher Scientific, Nalgene, 5100-0001)
- 1 ml带滤芯的移液枪头 (Thermo Fisher Scientific, catalog number: TF112-1000-Q)
- 75%酒精消毒液 (ANNJET, 75%酒精消毒液)
- 胎牛血清 (Cellmax, SA301.02)
- Schneider果蝇培养基 (Thermo Fisher Scientific, Gibco, catalog number: 21720-024)
- 青链霉素混合液 (Thermo Fisher Scientific, Gibco, catalog number: 15140122)
仪器设备
- 二级生物安全柜 (Esco, Airstream A2)
- 细胞培养箱 (SANYO, MCO-18AIC):有恒温和保湿功能即可,温度设定为25 °C,湿度设定为70%,果蝇S2细胞培养不需要通入CO2气体
- 真空泵 (Millipore, WP6122050)
- 倒置荧光显微镜 (Leica, DM IL LED)
- 移液枪 (Drummond, Pipet-Aid XP)
- 移液器 (Gilson, model: P1000)
- 恒温水浴锅 (Thermo Fisher Scientific, ISOTEMP 202)
- -80 °C超低温冰箱 (Esco, Lexicon, UUS-668B-1)
- 4 °C及-20 °C冰箱 (Haier, BCD-539WT)
实验步骤
Copyright: © 2019 The Authors; exclusive licensee Bio-protocol LLC.
您对这篇实验方案有问题吗?
在此处发布您的问题,我们将邀请本文作者来回答。同时,我们会将您的问题发布到Bio-protocol Exchange,以便寻求社区成员的帮助。
发布问题 0 Q&A