Collecting and Fixing Nuclear GFP/RFP in L1 Larva for Imaging
用来拍照的转化GFP/RFP的幼期蠕虫的收集和固定方法
发布: 2012年03月05日第2卷第5期 DOI: 10.21769/BioProtoc.94 浏览次数: 10753
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Abstract
In this protocol, L1 stage larvae are collected that carry nuclear-localized GFP/wCherry reporters. These can be fixed so that the GFP/wCherry maintains nuclear localization and stain nuclei by DAPI. This protocol therefore achieves the collection and fixation of nuclei in worm L1 larvae.
Materials and Reagents
- Acetone
- Formaldehyde
- DAPI
- Poly-lysine
- Glycerol
- Cytoseal 280 (Richard-Allan Scientific, catalog number: 8311-4 )
- Used Qiaquick Spin Column (QIAGEN)
- 11.58 μm glassbeads (Whitehouse Scientific, catalog number: MS0012 )
- KCl
- NaCl
- Na2EGTA
- Triton X-100
- EDTA
- PIPES
- 2x modified MRWB (see Recipes)
- DAPI staining solution (see Recipes)
- M9 buffer (see Recipes)
- Tris triton buffer (TTB) (see Recipes)
Equipment
- Standard bench top microcentrifuge
- 16-slide glass staining jar (Thermo Fisher Scientific, catalog number: 08-810 )
- Spatula
- Microscope
- Glass container
- Glass coverslip
- Glass slide
- 18 x 18 mm glass cover slip
- 25 x 75 mm glass slide
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Liu, X. (2012). Collecting and Fixing Nuclear GFP/RFP in L1 Larva for Imaging. Bio-protocol 2(5): e94. DOI: 10.21769/BioProtoc.94.
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分类
细胞生物学 > 细胞成像 > 荧光
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