A simple,chisel-assisted mechanical microdissection system for harvesting homogenous plant tissue suitable for the analysis of nucleic acids and proteins |
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Authors: | Stephan Brandt Christina Walz Martina Schad Nada Pavlovic Julia Kehr |
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Affiliation: | 1.Institut für Botanik,Technische Universit?t Darmstadt,Darmstadt,Germany;2.Department Willmitzer,Max Planck Institute of Molecular Plant Physiology,Golm,Germany;3.Eppendorf AG,Hamburg,Germany |
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Abstract: | Many physiological processes are limited to specific tissues or even specific cell types. Analysing entire plants or organs results in averaged data of all cell types contained in the sample; thus, specific metabolic functions cannot be assigned to individual cell types. A higher spatial resolution is required. By microdissecting plant organs, homogeneous material can be obtained. If a suitable amount of material is collected, standard analytical methods can be applied to elucidate cell type-specific processes. The collection of sufficient quantities of homogeneous material can be done by means of mechanical microdissection. This technique is a low-cost alternative to laser-coupled microdissection techniques. Here we describe a protocol for chisel-assisted mechanical microdissection of embedded plant material and demonstrate that the collected material is suitable to obtain nucleic acids and proteins. |
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Keywords: | mass spectrometry mechanical microdissection paraffin embedding protein sequencing RT-PCR single-cell analysis tissue specificity |
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