胚胎油体等球状体大小、分布的成像和定量测定

Martine Miquel; Ghassen Trigui; Alain Trubuil; Bertrand Dubreucq

doi:10.21769/BioProtoc.1369

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Peer-reviewed

Imaging and Quantitative Analysis of Size and Distribution of Spherical Bodies, e.g. Embryonic Oil Bodies

胚胎油体等球状体大小、分布的成像和定量测定

Martine Miquel email

GT Ghassen Trigui

AT Alain Trubuil

Bertrand Dubreucq email

发布: 2015年01月05日第5卷第1期 DOI: 10.21769/BioProtoc.1369 浏览次数: 8693

评审: Renate WeizbauerArsalan DaudiAnonymous reviewer(s)

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参见作者原研究论文

The authors used this protocol in:

Cover of Plant Physiology, featuring study using the protocol.

Apr 2014

实验方案合集

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Abstract

Oil bodies (OBs) are seed-specific lipid storage organelles that allow the accumulation of neutral lipids that sustain plantlet development after the onset of germination. Using fluorescent dyes and confocal microscopy, we monitored the dynamics of OBs in living Arabidopsis (Arabidopsis thaliana) embryos at different stages of development (Miquel et al., 2014). Image acquisition was followed by a detailed statistical analysis of OB size and distribution during seed development in the four dimensions (x, y, z, and t).

Keywords: Oil body (油体)

Embryo (胚胎)

Size (尺寸)

Imaging (成像)

Statistical analysis (统计分析)

Materials and Reagents

Plant material
1. Arabidopsis thaliana plants, wild type or mutant or transgenic plants.
2. Developing siliques between 6 and 11 days after pollination of plants grown in a greenhouse under the following conditions (13 h of light, diurnal temperature of 25 °C, and nocturnal temperature of 17 °C), and irrigated twice per week with mineral nutrient solution.
Nile Red (a neutral lipid stain, at 2 μg/ml final in acetone) (Sigma-Aldrich, catalog number: 72485 )

Equipment

Confocal microscope
Note: In this study an inverted LEICA SP2-AOBS spectral confocal laser microscope (Leica Microsystems) equipped with an HCX PL APO CS 40 x 1.25 objective and a multiline argon laser was used.
Forceps (Dumont No.5) (Sigma-Aldrich)
Scalpel (11 P, blade) (Swann Norton)
Glass slides or glass-bottom dish, cover slips
Binocular (Nikon Corporation, model: SMZ1000 ) (Champigny sur Marne with a led ring - Shott easyled^TM for illumination)

Software

ND-SAFIR (http://serpico.rennes.inria.fr/doku.php?id=software:nd-safir:index)
AVIZO^® FIRE (http://www.vsg3d.com/avizo/fire)

Procedure

English

中文翻译

文章信息

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如何引用

Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

Miquel, M., Trigui, G., Trubuil, A. and Dubreucq, B. (2015). Imaging and Quantitative Analysis of Size and Distribution of Spherical Bodies, e.g. Embryonic Oil Bodies. Bio-protocol 5(1): e1369. DOI: 10.21769/BioProtoc.1369.
Miquel, M., Trigui, G., d'Andrea, S., Kelemen, Z., Baud, S., Berger, A., Deruyffelaere, C., Trubuil, A., Lepiniec, L. and Dubreucq, B. (2014). Specialization of oleosins in oil body dynamics during seed development in Arabidopsis seeds. Plant Physiol 164(4): 1866-1878.