Application of laser microdissection to study plant-fungal pathogen interactions

Laser microdissection (LM) has become an important tool for isolating individual cells or cell types from suitably prepared tissue samples. The technique can be used to isolate both fungal and host plant cells after pathogen infection for molecular studies. Sample preparation is a crucial step in LM and involves fixing samples with appropriate fixatives to preserve the integrity of cell morphology and target metabolites (e.g., RNA), and embedding the fixed tissue in paraffin wax for sectioning onto microscope slides. The sample sections are then deparaffinised, rehydrated, and cells are dissected by a laser focused through a microscope. LM samples are collected into protective (e.g., RNAse-free) medium for isolation of RNA. The RNA can then be subjected to gene expression studies such as quantitative RT-PCR and microarray analysis after a linear RNA amplification process.