The laser in the Lowry technique for microdissection of freeze-dried tissue slices

Abstract

A new technique for tissue microdissection is described. This procedure, using an u.v.-laser micropreparation instrument, overcomes the extremely time-consuming manual preparation. The u.v.-laser micropreparation design allows fast, precise, reproducible and smear-contamination-free tissue microdissection. The preparation of the tissue sample can be programmed by tracing out the area to be sampled with a non-destructive 0.5 mW He-Ne-laser aiming-beam. The tract is stored in a small electronic unit, which then guides the motor-driven stepping stage on the microscope in the actual dissection run with the u.v.-laser. The laser power is adjustable in the range 4 to 40 kW and controlled by a photo diode displayed on an oscilloscope screen. In the tissue slice, prepared according to Lowry, an unlimited number of cells or tissue compartments can be dissected and afterwards weighed. The procedure described offers a broader use of the quantitative microhistochemical techniques of Lowry and of Neuhoff.

A strategy to identify genomic expression at single-cell level or a small number of cells

“…The use of lasers in tissue microdissection was reported as early as 1976 (Meier-Ruge et al 1976), and it has increasingly been applied in single-cell microdissection in recent years. In contrast to flow-cytometric cell sorting, microdissection allows for both rapid in vivo localization and the ability to analyze the cellular microenvironment (Schutze and Lahr, 1998).…”

Microgenomics: Identification of new expression profiles via small and single‐cell sample analyses

“…Originally, microdissection meant manually scraping off an area of interest from a sectioned tissue by the means of fine needles; later, micromanipulators were used in essentially the same way (21), only slightly improving the tedious, potentially inaccurate, and highly operator-dependent method of manual microdissection (22). The first laser-assisted microdissection method was published by Shibata et al (23), and was a method of negative selection employing ablation of unwanted tissue areas by selective ultraviolet radiation fractionation.…”

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.…”