Cell surgery by laser micro‐dissection: A preparative method

Abstract

A new UV-laser device is introduced as a micro-manipulator and micro-dissection apparatus for experimental cell research. High energy irradiation is produced by a quasi-continuous Nn-laser. The laser micro-beam can be focused to a minimal diameter of 0.75 pm. The micro-laser can be used for cutting and, hence, as a micro-surgical instrument at the level of cellular organelles.Cytoplasmic actomyosin fibrils (AM fibrils) from tissue culture cells (RMCDcells) are isolated from the surrounding ground cytoplasm. It is shown that the contractile proteins are not damaged by the laser dissection method to an extent that would prevent their normal function, e.g. contraction.

Application of Laser Microdissection to plant pathogenic and symbiotic interactions

“…This procedure is performed in a manner that is conducive to the extraction of DNA, RNA, proteins and even metabolites. 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).…”

Proteomic Analyses of Cells Isolated by Laser Microdissection

“…Thus, the primary reason for obtaining gene expression information from specific cell types is to minimize the dilution effect caused by the cellular complexity found in tissues and organs. This limitation has been overcome by the laser microdissection (LM) technique which was first described by Isenberg et al (1976) and then developed at the NIH (National Institute of Health, U.S.) for the dissection of cells from histological tissue sections (Emmert-Buck et al, 1996). Laser microdissection permits the rapid procurement of selected cell populations from a section of heterogeneous tissue in a manner conducive to the extraction of DNA, RNA or proteins.…”

“…However, although it is fairly easy to assay a transcriptome and a proteome at these higher levels of biological organization, assaying a proteome at a cellular resolution level involves a set of problems that in primarily centered on the ability to collect sufficient cells for meaningful studies. The central focus of this chapter is a discussion on the technologies that have allowed the proteomic analyses of cells, isolated from complex samples thanks to a procedure that was first called laser microdissection (Isenberg et al, 1976).…”

The use of laser capture microdissection to study the infection ofGlycine max(soybean) byHeterodera glycines(soybean cyst nematode)

“…LCM is a precise and accurate alternative to hand dissections for isolating homogeneous cell populations that are otherwise recalcitrant to their isolation. 23,24 Thus, LCM provided an opportunity to study the syncytium as an emergent property of the development of infection between G. max and H. glycines, by allowing the construction of cDNA libraries, cloning of genes and expression analyses. 25 It also allowed the isolation of homogeneous syncytium samples so that microarray analysis of syncytia undergoing a compatible or incompatible reaction could be performed.…”