Thin Sections of Technovit 7100 Resin of Rice Endosperm and Staining
采用Technovit 7100树脂制备水稻胚乳薄切片并染色
发布: 2014年09月20日第4卷第18期 DOI: 10.21769/BioProtoc.1239 浏览次数: 15027
评审: Tie LiuAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Feb 2014
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Abstract
Starch is a biologically and commercially important carbohydrate that is accumulated in plant storage organs, such as seed endosperm. Starch is water-insoluble and forms transparent grains, referred to as starch grains (SGs). SGs are easily stained by iodine solution and can be observed under a normal microscope. Technovit 7100 resin is suitable for preparation of thin sections from endosperm. The thin sections and iodine staining can visualize SGs clearly inside the endosperm cell. This protocol provides the procedures to prepare thin sections of Technovit 7100 resin from rice endosperm. It can also be applicable to the seeds of other plant species.
Figure 1. Iodine-stained Technovit thin sections of matured endosperm cells in rice and barley. Starch grains (SGs) can be visualized as violet particles.
Materials and Reagents
- Rice seeds
- Technovit 7100 resin (Heraeus Kulzer)
- Lugol solution (MP Biomedicals, catalog number: 155269 )
- Formalin
- Acetic acid
- 99.5% ethanol (commercially available one)
- Instant adhesives (Aron alpha, Toagosei Co.)
- FAA solution (see Recipes)
- 100% ethanol (see Recipes)
- 100% Technovit 7100 (I) (see Recipes)
- 25% Technovit 7100 resin (I) (see Recipes)
- 50% Technovit 7100 resin (I) (see Recipes)
- 75% Technovit 7100 resin (I) (see Recipes)
- 100% Technovit 7100 (II) (see Recipes)
Equipment
- Silanized slides (Dako, catalog number: S3003 )
- Wooden dowels (6 x 25 mm)
- Razor blade (Feather, catalog number: FH-10 )
- 1.5 ml plastic tube
- 0.2 ml PCR tube
- Molecular sieves 4A 1/16 (Nacalai tesque, catalog number: 04172-95 )
- Ultramicrotome (Leica Microsystems, model: EM UC7 )
- Diamond knife (Diatome AG, Ultra trim, 3.0 mm)
- Rotator (TAITEC, model: RT-30mini )
- Forceps (Vigor, stainless steel #5)
- Aspirator (Ulvac, model: MDA-015 )
- Microscope (OLYMPUS, model: AX70 )
Figure 2. A wooden dowel
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Matsushima, R. (2014). Thin Sections of Technovit 7100 Resin of Rice Endosperm and Staining. Bio-protocol 4(18): e1239. DOI: 10.21769/BioProtoc.1239.
- Matsushima, R., Maekawa, M., Kusano, M., Kondo, H., Fujita, N., Kawagoe, Y. and Sakamoto, W. (2014). Amyloplast-localized SUBSTANDARD STARCH GRAIN4 protein influences the size of starch grains in rice endosperm. Plant Physiol 164(2): 623-636.
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分类
植物科学 > 植物细胞生物学 > 组织分析
植物科学 > 植物细胞生物学 > 细胞成像
生物化学 > 糖类 > 多糖
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