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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Abbreviations
- LCM:
-
laser capture microdissection
- m/z:
-
mass/charge
- PPMD:
-
Piezo Power Microdissection
- Q-TOF:
-
quadrupole-time-of-flight
- RT-PCR:
-
reverse transcription-polymerase chain reaction
- RuBisCo:
-
ribulose-1,5-bisphosphate carboxylase
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
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Brandt, S., Walz, C., Schad, M. et al. A simple, chisel-assisted mechanical microdissection system for harvesting homogenous plant tissue suitable for the analysis of nucleic acids and proteins. Plant Mol Biol Rep 21, 417–427 (2003). https://doi.org/10.1007/BF02772591
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DOI: https://doi.org/10.1007/BF02772591