LASER MICRODISSECTION OF PLANT TISSUE: What You See Is What You Get

Abstract

Laser microdissection (LM) utilizes a cutting or harvesting laser to isolate specific cells from histological sections; the process is guided by microscopy. This provides a means of removing selected cells from complex tissues, based only on their identification by microscopic appearance, location, or staining properties (e.g., immunohistochemistry, reporter gene expression, etc.). Cells isolated by LM can be a source of cell-specific DNA, RNA, protein or metabolites for subsequent evaluation of DNA modifications, transcript/protein/metabolite profiling, or other cell-specific properties that would be averaged with those of neighboring cell types during analysis of undissected complex tissues. Plants are particularly amenable to the application of LM; the highly regular tissue organization and stable cell walls of plants facilitate the visual identification of most cell types even in unstained tissue sections. Plant cells isolated by LM have been the starting point for a variety of genomic and metabolite studies of specific cell types.

Ontogeny of the Maize Shoot Apical Meristem

“…Laser microdissection enables the isolation of discrete domains within microscopic samples for use in transcriptomic analyses (Nelson et al, 2006;Scanlon et al, 2009). Six samples were microdissected from developing embryos and 14-d-old seedlings, including (1) the cells comprising the embryo proper of the proembryo ( Figures 1M and 1S), (2) the organizing SAM and emerging scutellar hood of the transition-stage embryo ( Figures 1N and 1T), (3) the SAM and the initiating coleoptile of the coleoptile-stage embryo ( Figures 1O and 1U), (4) the SAM and leaf primordium of a stage 1 embryo ( Figures 1P and 1V), (5) the SAM and plastochron 1 leaf of a 14-d-old seedling (L14; Figures 1Q and 1W), and (6) the lateral meristem and the newly initiated husk leaf from the 14-d-old seedling ( Figures 1R and 1X).…”

Using Genomics to Study Legume Seed Development

“…One way to overcome the limitations of hand dissection and to be able to isolate different regions from any legume seed and embryo regardless of size (Fig. 2) is to make use of spectacular progress in laser capture microdissection (LCM) technology (Day et al, 2005;Nelson et al, 2006). LCM technology makes it possible to study gene activity in the entire seed because any seed compartment, region, or tissue can be isolated easily throughout development (Fig.…”

Application of Laser Microdissection to plant pathogenic and symbiotic interactions

“…LM has been widely used by animal biologists (Isenberg et al 1976) to study gene expression in specific cell types (Emmert-Buck et al 1996) and to elucidate the associated molecular events (reviewed in Fink and Bohle 2005;Espina et al 2006Espina et al , 2007. In more recent years, LM has been adapted for plant tissues (Day et al 2005(Day et al , 2006Nelson et al 2006;Ramsay et al 2006;Balestrini and Bonfante 2008). The advantage that this technology offers, with respect to other techniques that isolate homogeneous populations of a specific cell-type (i.e.…”

“…RNA, DNA, proteins, metabolites). A second problem is that the number of cells that can be recovered is limited by the abundance and recognition of the cellular targets in the histological sections (Nelson et al 2006). Another limitation is that LM instrumentation is costly.…”