We are currently working on developing two new technologies;expression microdissection (xMD) and layered expression scanning (LES). Based on immuno-targeting, xMD converts microdissection from an operator-dependent to an operator-independent mode, thus eliminating the need to laboriously procure cells. When fully developed xMD likely will offer several advantages over current methods, including;a) increased dissection rate (several orders of magnitude), b) increased dissection precision (subcellular), c) removal of variance among individual operators, permitting standardization of the dissection process, and d) elimination of targeting difficulties due to poor image quality of non-coverslipped (non-index-matched) histology sections used in current dissection systems. Layered expression scanning (LES) is a new technology co-developed by the Pathogenetics Unit and 20/20 GeneSystems, Inc. (www.2020gene.com/) through a Cooperative Research and Development Agreement (CRADA). The method utilizes a layered array of membranes for high-throughput molecular analysis and can be applied to a variety of life science platforms, including tissue sections, cells in culture, electrophoresis gels, multi-well plates, and tissue arrays. The technique is performed by passing the sample through a series of membrane layers while maintaining the original two-dimensional architecture. Each membrane measures a different molecular species, thus multiple RNA transcripts or proteins in each sample element (e.g., various cellular phenotypes in a tissue section, bands on a gel, individual wells of a microtiter plate) can be simultaneously analyzed. The method is conceptually simple, requires no moving parts, and can be used as an open or closed format.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010729-08
Application #
8763190
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
8
Fiscal Year
2013
Total Cost
$340,342
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Rodriguez-Canales, Jaime; Hanson, Jeffrey C; Hipp, Jason D et al. (2013) Optimal molecular profiling of tissue and tissue components: defining the best processing and microdissection methods for biomedical applications. Methods Mol Biol 980:61-120
Yan, Wusheng; Shih, Joanna; Rodriguez-Canales, Jaime et al. (2013) Three-dimensional mRNA measurements reveal minimal regional heterogeneity in esophageal squamous cell carcinoma. Am J Pathol 182:529-39