Since January a Leica LMD7000 Laser Microdissection (LMD) System is available for single cell isolation for multiple applications. The system is a new facility of CAT-AgroFood, Shared Research Facilities of Wageningen UR and is available for all interested researchers from Wageningen UR and other organisations.
To understand biological processes it is becoming increasingly important to analyse biomolecules at the resolution of single cells or cell-types. This Leica LMD7000 Laser Microdissection (LMD) System is a tool for this since it allows the contamination-free isolation of specific cells or subcellular structures for subsequent extraction of biomolecules (DNA, RNA, protein, metabolites) from almost any biological system. The LMD7000 uses a laser to cut out cells without heating or damaging surrounding tissue and collects dissected cells by gravity. In this way, cells can be isolated from either fixed or frozen tissue sections or from living cells (for example cell cultures) and fluorescence can be used to aid the selection of relevant cells. After isolation of the relevant cells downstream analyses can be performed such as genotyping, expression analyses, protein and metabolite analyses. The system can also be used to isolate specific living cells from cell cultures for subsequent sub-culturing and for cell-ablation or microsurgery in living tissue. Additionally, the system may be used for microengraving of slides or samples for electron microscopy.
Erik Limpens, Assistant Professor at the Laboratory of Molecular Biology at Wageningen UR, is the daily supervisor of the instrument and one of the users. He and his colleagues will use the system in their studies on the symbioses between plants and nitrogen-fixing rhizobium bacteria and arbuscular mycorrhizal fungi. During these interactions the microbes are accommodated inside specialized plant cells where both partners exchange nutrients, and in this way the plant can gets access to scarce minerals in the soil such as phosphate and fixed nitrogen. To understand how plant cells are reprogrammed to host the microbes, cells at different stages of the interactions will be isolated and used for genome-wide transcriptome analyses to identify key genes from both plant and microbe. In addition, individual nuclei will be isolated from mycorrhizal hyphae to study the genetic make-up of these ecologically important fungi and the influence of different plant hosts on its composition.