Thanks to everyone who attended the Arcturus LCM Users Meeting at MD Anderson Cancer Center in Houston, TX. Special thanks to all of the speakers for helping to make this user meeting a great success.
The presentation topics are listed below and available for viewing in PDF format. If a presentation is not available here, please contact the author directly for more information.
- Transcription Signatures of Metastatic Cancer
Dr. Phillip Febbo – Duke Institute for Genome Science and Policy, Departments of Medicine and Molecular Genetics and Microbiology
Metastatic cancers kill over 500,000 people annually in the United States. In order to investigate the biology differentiating between metastatic cancer from different primary sites to different metastatic locations, we performed laser capture microdissection, RNA amplification, and Affymetrix microarray analysis on 103 metastatic tumors. Analysis of replicate samples demonstrated that the variation between replicates was significantly less that that between different cancers supporting our prior findings that the technical variation introduced by LCM and RNA amplification does not obscure intra-tumoral differences. Unsupervised and supervised analysis suggested that the persistent biological signature derived from the site of origin predominates the global gene expression patterns for these samples. However, metastatic-site address signatures can be developed that accurately predict the location of independent validation samples of metastatic cancer. Finally, using a number of recently developed computational approaches we found specific biological processes associated site of metastasis.
- Presentation not available. Please contact author for more information.
- Application of Laser Capture Microdissection and ‘Micro-omics’ Analyses to Toxicology and Environmental Health Effects Research
Dr. Heather Flyod and Dr. Kevin Dreher, Experimental Toxicology Division, National Health and Environmental Effects Laboratory, US Environmental Protection Agency, Research Triangle Park, NC
The presentation by Drs. Floyd and Dreher will present fundamental background information on laser capture microdissection as well as methods and applications of this technology for in situ molecular toxicology (toxicogenomics and proteomics) for environmental health risk assessment research. The presentation will stress techniques and methods for tissue preparation as well as to identify regions of interest within tissue samples for laser capture microdissection. In addition, this lecture will briefly describe subsequent downstream functional micro-genomic or proteomic analyses that can be performed on as little as 500 or less cells. Information presented will allow for specific and direct in vivo assessment of molecular toxicity as well as provide a means to extrapolate or validate in vitro toxicological methods as well as enhance the ability to detect adverse biological effects at lower exposure doses. Application of LCM and molecular profiling results will be presented from the following studies:
- Alteration in airway intracellular signaling pathways following air pollution particle (PM) exposure using laser capture microdissection and protein array technologies
- Detection and molecular analysis of particulate air pollution-induced cardiopulmoonary oxidative stress using a transgenic mouse model, laser capture microdissection, and Toxicogenomics
- Fine Ambient Air Particular Matter Exposure Induces Molecular Alterations Indicative of Cardiovascular Disease Progression in Atherosclerotic Susceptible Mice
- Laser Capture Microdissection in Gene Expression Studies of Songbird Brains
Osceola Whitney and Dr. Miriam Rivas – Duke University Department of Neurobiology
- Microarray Applications: Gene Expression and Beyond
Eric Levy – Application Scientist, MDS Analytical Technologies
Microarrays began essentially as a rapid method of performing several Northern blots for many genes at once. From their humble beginnings as "slot blot" for gene expression, microarrays have moved into a multitude of applications, including genetic analysis at the subchromosomal all the way to SNP levels, proteomics, methylation, "interactomics," and beyond. With the increasingly robust commercial productions of arrays, perfection of substrates for printing, and advancement in analytical software, microarray promises to become an ever more ubiquitous tool in a variety of labs, from clinical to basic research.
- Arcturus LCM and Microgenomics Product Overview
David Choiniere, MDS Analytical Technologies
Laser Capture Microdissection (LCM) is a proven technique for isolating pure cell populations for downstream molecular analysis. Combined UV laser cutting and LCM using the IR laser, found only with Arcturus® LCM Instruments, allows for rapid and precise isolation of larger numbers of cells, while maintaining cellular and nucleic acid integrity necessary for downstream analysis.
Molecular Devices, a Division of MDS Analytical Technologies, recently introduced the ArcturusXT™ Microdissection Instrument. The open-architecture ArcturusXT system is built upon the Nikon TE2000U® inverted research grade microscope, and combines infrared (IR) laser-enabled laser capture microdissection (LCM) and ultraviolet (UV) laser cutting in a single modular instrument, offering researchers superior speed, precision and flexibility. By utilizing the Nikon TE2000U, the ArcturusXT is completely modular and fully upgradeable, allowing the system utility to expand as research requirements grow and microdissection technologies emerge.
Molecular Devices offers the full solution of Microgenomics reagent products that support and complement the LCM technology, from kits for tissue section staining and nucleic acid extraction to kits for RNA amplification and labeling for microarray analysis. The microgenomics platform benefits the researcher as it ensures high efficiency recovery of quality total RNA from as little as a single cell, obtained through LCM. High sensitivity linear amplification of mRNA is possible from as little as 100 picograms (10 LCM cells) of total RNA, generating enough aRNA for replicate microarray hybridizations. Linear amplification is followed by non-enzymatic labeling, which allows the use of unlabeled nucleotides during amplification. This enables the use of lesser amounts of starting material and results in higher aRNA yields and higher %P calls during microarray analysis. This complete microgenomics platform has been validated for use with all common microarray platform
- Arcturus LCM Protocols and Troubleshooting
Scott Mordecai, Senior Application Scientist, MDS Analytical Technologies
