How does LCM work?
Laser Capture Microdissection is a new technology developed to enable investigators and clinicians to perform tissue microdissection on a routine basis. The instrument was conceived as a prototype research tool at the NIH. It was then developed through a cooperative research and development agreement (CRADA) between Arcturus Engineering and NIH, NCRR, and NCI.
LCM provides a rapid, reliable, one step method to procure pure populations of targeted cells from specific microscopic regions of tissue sections for subsequent analysis. A thermoplastic polymer used on a cap (see diagram below) bonds to the tissue after being targeted with the laser. This avoids chemical reactions that can cross-link biological molecules in the tissue and alter subsequent molecular analysis. LCM has successfully extracted cells in all tissues in which it has been tested. In addition, there are no limitations in the ability to amplify DNA or RNA from tumor cells. LCM can capture cells without damaging the enzymatic function of the proteins.
Extraction of DNA has been demonstrated using both ethanol fixed and formalin fixed paraffin embedded tissue. RNA can be recovered by dissecting fresh frozen tissue sections. Some guidelines for tissue needed for molecular analysis: 10-20 cells from a 10-micron thick PET section has been shown to work for PCR, 50,000 cells for a 2-D gel analysis and 20,000 cells for Western Blot. LCM has been used to explore gene expression, loss of heterozygosity, micro-satellite instability, clonality and in proteomics.
The LCM instrument is available for your use in the Nash Annex, Room 250F. Please contact Dr. Hao Chen @ 448-3720 for information on training sessions.
Laser Capture Microdissection Facility
Hao Chen, Ph,D.
Department of Pharmacology
Nash Annex, Room 250F
Memphis, TN 38163