- Jianming Pei, MD
- CPP P3011
The state-of-the-art technology of laser capture microdissection (LCM) provides researchers with the ability to accurately analyze DNA, RNA and protein from pure populations of cells, such as tumor cells that are captured from complex heterogeneous tissue samples. Protocols have been developed and optimized for acquisition of high-quality RNA and DNA from both frozen and formalin-fixed, paraffin-embedded tissue. LCM services include slide preparation and staining, and RNA and DNA extraction with qualitative analysis by Bioanalyzer or Nanodrop systems. Care is taken to preserve the integrity of the samples at all times, ensuring high quality retrieval of molecular targets.
Many different types of molecular analyses have been successfully performed on cells procured by LCM from frozen sections or paraffin-embedded tissues. These include:
1. Genomic analyses, such as:
a. Loss of heterozygosity analysis
b. DNA methylation analysis
c. Fluorescence in situ hybridization
d. Comparative genomic hybridization
2. Gene expression analysis (i.e., RNA analysis)
3. Proteomic analysis
LCM is equipped with two LCM systems (PixCell II® and Veritas™, Molecular Devices, Sunnywale, CA), which are available for a wide range of applications, including the LCM of specific immunostained or fluorescent cells. Viability of the IR laser-enabled LCM technique has been shown for a variety of different cell types. The LCM procedure is remarkably simple and robust. The method is extraordinarily gentle and ideal for microdissection of single cells or small numbers of cells. LCM system utilizes a laser microbeam that melts a thermoplastic membrane that sticks to the selected cells, which can then be lifted and secured in a microfuge tube containing the appropriate extraction solutions. The transferred tissue on the film retains its original morphology, thereby allowing microscopic verification of the specificity of the captured material. Using this strategy, a single small cluster up to thousands of cells can be procured safely and without contamination. The Agilent 2100 Bioanalyzer and Nanodrop ND1000 monitor the quality and quantity of extracted RNA, DNA, as well as proteins from the LCM sample.