树脂包埋制备拟南芥横切片

Shweta Kalve; Kumud Saini; Kris Vissenberg; Tom Beeckman; Gerrit T. S. Beemster

doi:10.21769/BioProtoc.1592

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
CN
- EN - English
- CN - 中文

Peer-reviewed

Transverse Sectioning of Arabidopsis thaliana Leaves Using Resin Embedding

树脂包埋制备拟南芥横切片

GB Gerrit T. S. Beemster email

(*contributed equally to this work) 发布: 2015年09月20日第5卷第18期 DOI: 10.21769/BioProtoc.1592 浏览次数: 13441

评审: Maria SinetovaMoritz BomerRumen Ivanov

下载 PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Journal of Experimental Botany, featuring study using the protocol.

Dec 2014

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

活体植物根部细胞核的近红外自发荧光成像

Akira Yoshinari and Masayoshi Nakamura

2025年04月20日 1030 阅读

利用活细胞成像系统研究拟南芥珠被中淀粉体的复制

Makoto T. Fujiwara [...] Ryuuichi D. Itoh

2025年06月05日 1366 阅读

活体叶片切片成像观察细胞内叶绿体运动及细胞间相互作用

Yuta Kato [...] Mitsutaka Taniguchi

2025年08月05日 968 阅读

Abstract

The leaf is the major functional part of the shoot performing the bulk of photosynthetic activity. Its development is relatively plastic allowing the plant to adapt to environmental changes by modifying leaf size and anatomy. Moreover, a leaf is made up of various distinct cell layers, each having specialized functions. To understand functional adaptation and the development of the leaf it is essential to obtain cross sections throughout leaf development and at maturity (Kalve et al., 2014). Here, we describe a protocol for transverse sectioning of Arabidopsis thaliana leaves using resin embedding. This protocol provides a reliable platform to yield high quality images of cross sections allowing study of development of various tissue layers across the transversal axis of the leaf. As this method is an adaptation of the protocol developed for the Arabidopsis root tip by Beeckman and Viane (1999) and De Smet et al. (2004), it can easily be modified to accommodate other organs and species.

Keywords: Microscopy (显微镜)

Arabidopsis thaliana (拟南芥)

Leaves (叶)

Microtomy (切片)

Materials and Reagents

Arabidopsis thaliana
Acetic Acid, Glacial (Certified ACS), Fisher Chemical (Fisher Scientific, catalog number: A38-212 )
Formaldehyde solution (Sigma-Aldrich catalog, number: 47629 )
Ethanol (VWR International, catalog number: 83813360 )
Acetic anhydride (Sigma-Aldrich, catalog number: 320102 )
TWEEN^® 20 (Sigma-Aldrich, catalog number: P9416 )
NaH₂PO₄^.H₂O (Fisher Scientific, catalog number: AC2717500190 )
Technovit 7100 with hardener I and II (Heraeus Kulzer GmbH, catalog number: 64709003 )
Toluidine blue (EMD Millipore Corporation, catalog number: 1159300025 )
DPX mounting medium (a mixture of Distyrene, a Plasticizer and Xylene) (Sigma- Aldrich, catalog number: M1289-10 ML )
Object slides (superfrost plus adhesion slides) (Thermo Fisher Scientific, catalog number: 10143352 )
Glass microtome knives (SPI Easy-cut Ultra/Glass, catalog number: 07668-BA )
Cover slips
Microcentrifuge tubes, PCR tubes or molds (Electron microscopy sciences, catalog number: 70900 )
Phosphate buffer (see Recipes)
Technovit solution (see Recipes)
Technovit solution with hardener-II (see Recipes)

Equipment

Vacuum pump (Welch USA- BRS, catalog number: 2522C-02 )
Vacuum jar
Tweezers
Heated plate (Bekso, model: TH3N )
Rotary microtome (Leica Reichert-Jung, catalog number: 2040 )
Bright field microscope (ZEISS, model: Axio Scope A1 ) fitted with a digital camera (ZEISS, model: AxioCamCm1 )

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Kalve, S., Saini, K., Vissenberg, K., Beeckman, T. and Beemster, G. T. S. (2015). Transverse Sectioning of Arabidopsis thaliana Leaves Using Resin Embedding . Bio-protocol 5(18): e1592. DOI: 10.21769/BioProtoc.1592.