Hippocampal Neuron Dissociation Transfection and Culture in Microfluidics Chambers
海马体神经的分离转化以及在微腔中的培养
发布: 2012年07月20日第2卷第14期 DOI: 10.21769/BioProtoc.235 浏览次数: 16505
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参见作者原研究论文
The authors used this protocol in:
Dec 2006
Abstract
Microfluidics chamber is an ideal tool to study local events that occurring in neuronal projections by perfectly compartmentalizing the cell soma from certain branches. It is very well suited for live cell imaging or immunohistochemistry staining. This protocol has been carefully modified in detail to fit the requirement of primary rat hippocampal neuronal cultures. It can also be applied to a more general neuronal culture purpose in microfluidics.
Background
Materials and Reagents
- β-mercaptoethanol (Sigma-Aldrich, catalog number: M3148 )
- HBSS (Hyclone, catalog number: SH30268 )
- Boric acid (Sigma-Aldrich, catalog number: B0252 )
- Borax (Sigma-Aldrich, catalog number: B9876 )
- Cysteine (Sigma-Aldrich, catalog number: C7352 )
- EtOH
- Neural basal media (NBM)
- Glutamax
- B27
- PenStrep
- Gentamicin
- Glutamate
- FBS
- Laminin
- Trypan blue
- Na2SO4
- K2SO4
- MgCl2
- CaCl2
- Glucose
- Phenol Red
- EDTA
- 0.1 M borate buffer (pH 8.5) (see Recipes)
- Dissection media (DM) (see Recipes)
- Papain activation buffer (see Recipes)
- Papain (100 mg) (Worthington Biochemical Corp, catalog number: LS003119 ) (see Recipes)
- DNase (F. Hoffmann-La Roche, catalog number: 10104159001 ) (see Recipes)
- Papain digestion media (see Recipes)
- Dissociation media (see Recipes)
- Plating media with phenol red (see Recipes)
- Growth media without phenol red (see Recipes)
Equipment
- PDMS devices
- Microscope
- Hemacytometer
- TC hood
- Incubator
- -80 °C freezer
- 37 °C water bath
- 6 cm cell culture dishs
- Plasma-bonding machine (model PDC-32G , http://www.harrickplasma.com/)
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Geng, Y. (2012). Hippocampal Neuron Dissociation Transfection and Culture in Microfluidics Chambers. Bio-protocol 2(14): e235. DOI: 10.21769/BioProtoc.235.
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分类
神经科学 > 神经解剖学和神经环路 > 活细胞成像
发育生物学 > 细胞信号传导
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