Fluoro-Jade B Staining for Neuronal Cell Death
神经元细胞死亡的Fluoro-Jade B染色
发布: 2016年01月05日第6卷第1期 DOI: 10.21769/BioProtoc.1702 浏览次数: 24222
评审: Jia LiSabine Le SauxAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Apr 2015
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Abstract
Fluoro-Jade is a fluorescent derivative used for histological staining of degenerating neurons. This technique is simple and sensitive enough to label distal dendrites, axons, axon terminals as well as neuronal bodies. Fluoro-Jade has excitation and emission peak of 480 and 525 nanometer respectively. It can be visualized using a fluorescein/FITC filter. Some reports have demonstrated that Fluoro-Jade can also be useful to detect glial cell death (Anderson et al., 2013; Damjanac et al., 2007).
Keywords: Fluoro-Jade B (Fluoro-JadeB)
Brain staining (脑染色)
Cell death (细胞死亡)
Neurone (神经元)
Materials and Reagents
- Superfrost plus Microscope slide (Thermo Fisher Scientific, catalog number: 12-550-17 )
- Cover Glass (Thermo Fisher Scientific, catalog number: 12-545-88 )
- Tissue sample
- Fluoro-Jade B (Merck Millipore Corporation, catalog number: AG310 )
- Paraformaldehyde (Electron Microscopy Science, catalog number: 19210 )
- Potassium permanganate (KMnO4) (Sigma-Aldrich, catalog number: 223468 )
- DAPI (Life Technologies, catalog number: D3571 )
Note: Currently, it is “Thermo Fisher Scientific, Molecular ProbesTM, catalog number: D3571”. - Glacial Acetic acid (CH3CO2H) (Sigma-Aldrich, catalog number: A9967 )
- Ethanol
- Xylene (Sigma-Aldrich, catalog number: 534056 )
- Sodium hydroxide (NaOH) (Sigma-Aldrich, catalog number: S8045 )
- Sodium tetraborate decahydrate (Sigma-Aldrich, catalogue number: B9876 )
- Sodium chloride (NaCl) (Sigma-Aldrich, catalog number: S3014 )
- potassium phosphate dibasic (K2HPO4) (Sigma-Aldrich, catalog number: P3786 )
- potassium phosphate monobasic (KH2PO4) (Sigma-Aldrich, catalog number: P9791 )
- 4% paraformaldehyde (see Recipes)
- KPBS (see Recipes)
- 0.2% Fluoro-Jade (see Recipes)
- Fluoro-Jade solution (see Recipes)
- 0.2% DAPI (see Recipes)
Equipment
- Vacuum Desiccators (Thermo Fisher Scientific, catalog number: 08-642-5 )
- Tissue-Tek slide staining set (Electron Microscopy Science, catalog number: 62540-01 )
- 24 slide holder (Electron Microscopy Science, catalog number: 62543-06 )
- Orbital shaker
- Timer
- Slide Warmer
- DPX mounting medium (a mixture of the polystyrene distyrene and the plasticizer dibutylphthalate) (Electron Microscopy Science, catalog number: 13512 )
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Laflamme, N., Préfontaine, P. and Rivest, S. (2016). Fluoro-Jade B Staining for Neuronal Cell Death. Bio-protocol 6(1): e1702. DOI: 10.21769/BioProtoc.1702.
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分类
神经科学 > 发育 > 组织学染色
细胞生物学 > 细胞染色 > 全细胞
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