Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana

Martina Schad, Rajsree Mungur, Oliver Fiehn, Julia Kehr

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Background: Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents.Results: In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (~5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues.Conclusion: This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used.

Original languageEnglish (US)
Article number2
JournalPlant Methods
Volume1
Issue number1
DOIs
StatePublished - Aug 18 2005

Fingerprint

metabolomics
vascular bundles
Arabidopsis
Microdissection
lasers
Blood Vessels
Lasers
Arabidopsis thaliana
metabolites
Tissue Embedding
Cryoultramicrotomy
Tissue Fixation
Plant Cells
Cellular Structures
Gas Chromatography
sampling
Mass Spectrometry
RNA
animal tissues
cell structures

Keywords

  • GC-TOF MS
  • laser microdissection
  • metabolite profiling
  • tissue-specific analysis
  • vascular bundle

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Plant Science

Cite this

Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana. / Schad, Martina; Mungur, Rajsree; Fiehn, Oliver; Kehr, Julia.

In: Plant Methods, Vol. 1, No. 1, 2, 18.08.2005.

Research output: Contribution to journalArticle

Schad, Martina ; Mungur, Rajsree ; Fiehn, Oliver ; Kehr, Julia. / Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana. In: Plant Methods. 2005 ; Vol. 1, No. 1.
@article{14a013c3d8fc44cfbb192527a052f16f,
title = "Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana",
abstract = "Background: Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents.Results: In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (~5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues.Conclusion: This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used.",
keywords = "GC-TOF MS, laser microdissection, metabolite profiling, tissue-specific analysis, vascular bundle",
author = "Martina Schad and Rajsree Mungur and Oliver Fiehn and Julia Kehr",
year = "2005",
month = "8",
day = "18",
doi = "10.1186/1746-4811-1-2",
language = "English (US)",
volume = "1",
journal = "Plant Methods",
issn = "1746-4811",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana

AU - Schad, Martina

AU - Mungur, Rajsree

AU - Fiehn, Oliver

AU - Kehr, Julia

PY - 2005/8/18

Y1 - 2005/8/18

N2 - Background: Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents.Results: In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (~5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues.Conclusion: This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used.

AB - Background: Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents.Results: In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (~5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues.Conclusion: This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used.

KW - GC-TOF MS

KW - laser microdissection

KW - metabolite profiling

KW - tissue-specific analysis

KW - vascular bundle

UR - http://www.scopus.com/inward/record.url?scp=33745944637&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745944637&partnerID=8YFLogxK

U2 - 10.1186/1746-4811-1-2

DO - 10.1186/1746-4811-1-2

M3 - Article

AN - SCOPUS:33745944637

VL - 1

JO - Plant Methods

JF - Plant Methods

SN - 1746-4811

IS - 1

M1 - 2

ER -