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.

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National Cancer Institute (NCI)
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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
Blackler, A R; Morgan, N Y; Gao, B et al. (2013) Proteomic analysis of nuclei dissected from fixed rat brain tissue using expression microdissection. Anal Chem 85:7139-45
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
Roy Chowdhuri, Sinchita; Hanson, Jeffrey; Cheng, Jerome et al. (2012) Semiautomated laser capture microdissection of lung adenocarcinoma cytology samples. Acta Cytol 56:622-31
Yan, Wusheng; Shih, Joanna H; Rodriguez-Canales, Jaime et al. (2012) Identification of unique expression signatures and therapeutic targets in esophageal squamous cell carcinoma. BMC Res Notes 5:73
Hipp, Jason; Monaco, James; Kunju, L Priya et al. (2012) Integration of architectural and cytologic driven image algorithms for prostate adenocarcinoma identification. Anal Cell Pathol (Amst) 35:251-65
Chowdhuri, Sinchita Roy; Xi, Liqiang; Pham, Trinh Hoc-Tran et al. (2012) EGFR and KRAS mutation analysis in cytologic samples of lung adenocarcinoma enabled by laser capture microdissection. Mod Pathol 25:548-55
Hipp, Jason D; Lucas, David R; Emmert-Buck, Michael R et al. (2011) Digital slide repositories for publications: lessons learned from the microarray community. Am J Surg Pathol 35:783-6
Hipp, Jason; Cheng, Jerome; Hanson, Jeffrey C et al. (2011) SIVQ-aided laser capture microdissection: A tool for high-throughput expression profiling. J Pathol Inform 2:19
Nacev, Alek; Kim, Skye H; Rodriguez-Canales, Jaime et al. (2011) A dynamic magnetic shift method to increase nanoparticle concentration in cancer metastases: a feasibility study using simulations on autopsy specimens. Int J Nanomedicine 6:2907-23

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