扫描电子显微镜（SEM）观测橄榄花序的起始和发育

Amnon   Haberman; Einat  Zelinger; Alon  Samach

doi:10.21769/BioProtoc.2575

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
登录
/
注册
- 个人主页
- 编辑个人信息
- 修改密码
- 退出
CN
- EN - English
- CN - 中文

Peer-reviewed

Scanning Electron Microscope (SEM) Imaging to Determine Inflorescence Initiation and Development in Olive

扫描电子显微镜（SEM）观测橄榄花序的起始和发育

Amnon Haberman email

EZ Einat Zelinger

AS Alon Samach

发布: 2017年10月05日第7卷第19期 DOI: 10.21769/BioProtoc.2575 浏览次数: 8478

评审: Scott A M McAdamAnonymous reviewer(s)

PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Plant, Cell & Environment, featuring study using the protocol.

Jan 2017

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

白葵花分生组织的定量实时共聚焦成像

Ya Min [...] Elena M. Kramer

2022年06月20日 2845 阅读

改良的拟Schiff碘化丙啶对拟南芥茎尖分生组织的染色

Ruiqi Li [...] Ligeng Ma

2023年05月05日 1648 阅读

使用 X 射线研究拟南芥长角果和种子的新型成像方案

Brylie A. Ritchie [...] Ansul Lokdarshi

2023年10月05日 1590 阅读

Abstract

Here we present a protocol that describes how to image the structure of the olive axillary bud meristem with a scanning electron microscope (SEM) in order to characterize its identity and developmental stage. Briefly, the specimen is fixed with glutaraldehyde, saturated with ethanol, dried in a critical point dryer (CPD) system, dissected, coated with a conducting material and imaged with a scanning electron microscopy (SEM).

Keywords: SEM (SEM)

Scanning Electron Microscopy (扫描电子显微镜)

Background

The exact timing of flowering induction and inflorescence initiation in olive (Olea europaea L.) is in controversy (Haberman et al., 2017). In olive, inflorescences emerge from lateral buds at the end of winter and flower in the spring. We have developed a protocol to better characterize the timing of inflorescence initiation in olive by imaging the meristem in the olive bud with a SEM at different times during the year. In these SEM images the meristem structure can be identified unambiguously, and the definition level of the meristem can be much higher than images of bud meristem sections presented in previous studies.

Materials and Reagents

Scalpel blade No. 11 (Sigma-Aldrich, catalog number: S2771 )
Double-sided adhesive tape
Glass scintillation vials with screw caps, volume 20 ml (Sigma-Aldrich, catalog number: Z190535 )
Pipette (BRAND, catalog number: 747760 ) or a similar instrument
Gold annular target for the sputter coater (Agar scientific, catalog number: AGB7370 )
Olive (Olea europaea L.) buds
Sodium phosphate dibasic (Na₂HPO₄) (Sigma-Aldrich, catalog number: S3264 )
Sodium phosphate monobasic (NaH₂PO₄) (Sigma-Aldrich, catalog number: S3139 )
25% glutaraldehyde (Sigma-Aldrich, catalog number: G5882 )
Ethanol absolute (Sigma-Aldrich, catalog number: 24102 )
Optional: Technical grade ethanol (Sigma-Aldrich, catalog number: V0T0042 )
0.1 M phosphate buffer pH 7.2 (sodium phosphate buffer; see Recipes)
5% glutaraldehyde solution (in 0.1 M phosphate buffer pH 7.2; see Recipes)

Equipment

Scalpel handle No. 3 (Sigma-Aldrich, catalog number: S2896 )
Tweezers style #5 (Sigma-Aldrich, catalog number: T4537 )
Critical point dryer system (BAL-TEC, model: CPD 030 )
Stereo-microscope (Olympus, model: SZX12 )
Sputter coater (E510 scanning electron microscope coating unit) (Polaron Instruments, model: E510 )
Scanning electron microscope (JEOL, model: JSM-5410 LV )

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Haberman, A., Zelinger, E. and Samach, A. (2017). Scanning Electron Microscope (SEM) Imaging to Determine Inflorescence Initiation and Development in Olive. Bio-protocol 7(19): e2575. DOI: 10.21769/BioProtoc.2575.