Utilization of laser capture microdissection in diagnosis and prognosis of cancer disease

Laser capture microdissection (LCM) is a rapid, reliable method for obtaining pure populations of targeted cells from specific microscopic regions of tissue sections for subsequent analysis. LCM is based on the adherence of visually selected cells to a thermoplastic membrane, which overlies the dehydrated tissue section and is focally melted by the triggering of a low energy infrared laser pulse. Tissue sections are mounted on standard glass slides, and a transparent thermoplastic membrane is then placed over the dry section. The laser provides enough energy to transiently melt this thermoplastic film into the target cells. Several systems are available for LCM, and these vary in cell-capture method, system configuration and applications. LCM was applied to a wide range of cell and tissue preparations including frozen samples, formalin-fixed paraffin-embedded tissue or cytology smear. Depending on the starting material, DNA, good quality mRNA, and proteins can by extracted successfullyfrom captured tissue fragments, down to the single cell level. In combination with another techniques like expression library construction and cDNA array hybridisation, LCM will allow the establishment of new diagnostic and prognostic markers. This approach could help in establishing individualised a tailored to the molecular profile of a tumor.

This project was supported by grants MSM 6198959216, IGA NR/9076 and the MPO 1H-PK/45.