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Biology

通过飞沫标题细胞封装

doi: 10.3791/316 Published: October 1, 2007

Summary

Protocol

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NIH3T3细胞的准备:

A.弹出细胞

  1. Trypsinize细胞与细胞媒体,然后稀释1:1,从T75烧瓶中转移到一个15毫升的猎鹰管
  2. 分拆成颗粒细胞,离心,吸出上清液,用DPBS细胞
  3. 成颗粒细胞再次离心,吸上清
  4. 在媒体的悬浮细胞
  5. 确定细胞密度与血球(〜200 × 10 4细胞/ mL的容量瓶中每T75)
  6. 离心细胞液,吸出上清液,并悬浮在适量的媒体不同的细胞浓度

B.细胞弹射

  1. 弹射前使用的涡细胞
  2. 转移到注射器200μL细胞液
  3. 设置适当的模式脉冲发生器
    1. 弹出的单液滴和多个水滴(连发),设置脉冲发生器的“E BUR”模式
    2. 对于连续液滴抛射,至“NORM”模式设置脉冲发生器
  4. 变化的信号设置
    1. 设置高和低电平输出电压:5 V和LOL为0 V,并确保“林先生”LED是上的HIL
    2. 设置为一个方形脉冲信号
    3. 变化的时间液滴弹射电磁阀打开,改变为“妇女参与发展”的价值或改变占空比(“责任”)
    4. 改变改变“市盈率”价值弹射频率
    5. 更改价值水滴喷出一阵改变“BUR”
  5. 到准备基板上弹出电池解决方案与显微镜成像

C.染色

  1. 染料溶液与0.5μL,钙黄绿素- AM和2μL,乙锭每DPBS毫升二聚体
  2. 在染料溶液浸入准备基板
  3. 允许样品孵育10分钟,在37 °成像彗星之前

实验验证

  1. 在尼康的Eclipse TE - 2000 U荧光显微镜
    1. 现货先进的软件(诊断公司)
    2. 活/死含量

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Materials

Name Type Company Catalog Number Comments
Fluorescent Microscope Nikon Instruments Eclipse TE-2000 U
Solenoid valve cell ejector Operation frequency: 1 KHz, 30psi, 50nl~0.5ul. 12V Valve Driver: 2.5 Amp drive current
5-Gallon Portable air tank Coleman Powermate CT5 Pressurized air: 30 PSI
Pressure regulators Marsh Bellofram
Pulse Generator HP8112A Actuation frequency generation: 50 MHz
XYZ-stage Newmark Systems With Stepping motor: 6inch travel xy-stage(NLS4-6-25), 2.5inch travel z-stage(NLS4-2.5-25), 3axis controller(NSC-G3), 0.1um resolution
NIH 3T3 Cell-line fibroblasts
Trypsin 0.05% solution
NIH 3T3 cell medium
DPBS Buffer
T75 Tissue culture flasks
Plastic conical tubes 15 ml, for tissue culture

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通过飞沫标题细胞封装
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Cite this Article

Moon, S., Lin, P., Keles, H. O., Yoo, S., Demirci, U. Title Cell Encapsulation by Droplets. J. Vis. Exp. (8), e316, doi:10.3791/316 (2007).More

Moon, S., Lin, P., Keles, H. O., Yoo, S., Demirci, U. Title Cell Encapsulation by Droplets. J. Vis. Exp. (8), e316, doi:10.3791/316 (2007).

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