Proteohistography--Direct Analysis of Tissue with High Sensitivity and High Spatial Resolution Using ProteinChip Technology

Günther Ernst ¹, Christian Melle ¹, Bettina Schimmel ¹, Annett Bleul ¹ and Ferdinand von Eggeling ^1*

¹ Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, Jena, Germany

^* To whom correspondence should be addressed. E-mail: fegg{at}mti.uni-jena.de.

Submitted on June 20, 2005
Accepted on 11 August 2005

	Abstract

On the proteomic level all tissues, tissue constituents or even single cells are heterogeneous, but the biological relevance of this cannot be adequately investigated with any currently available technique. This is because the analysis of proteins of small tissue areas by any proteomic approach is limited by the number of required cells; increasing the number of cells only serves to lower the spatial resolution of expressed proteins. To enhance sensitivity and spatial resolution we developed Proteohistography (PHG). Laser microdissection was used to mark special areas of interest on tissue sections attached to glass slides. These areas were positioned under microscopic control directly on an affinity chromatographic ProteinChip Array so that cells were lysed and their released proteins bound on a spatially defined point. The ProteinChip^® System (SELDI-TOF-MS) allows to steer the laser to up to 215 distinct positions across the surface of the spot, enabling a high spatial resolution of measured protein profiles for the analyzed tissue area. Protein profiles of the single positions were visually plotted over the used tissue section to visualize proteohistologically distribution. Results show that the spatial distribution of detectable proteins could be used as Proteohistogram for a given tissue area. Consequently this procedure can provide additionally information to both a MALDI-based approach and immunohistochemistry, as it is more sensitive, highly quantitative and no specific antibody is needed. Hence, proteomic heterogeneity can be visualized even if proteins are not known or identified.

Key Words: ProteinChip arrays, SELDI, IHC, proteohistography, tissue, heterogeneity

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. Machaalani, M. Arlotto, K. A. Waters, E. Gozal, F. Berger, and M. Dematteis A Novel Method of Tissue Collection and Storage: Validation Using SELDI-TOF MS Analysis Clin. Chem., July 1, 2007; 53(7): 1387 - 1389. [Full Text] [PDF]

				H. Tonnies, J. Pietrzak, E. Bocian, K. MacDermont, A. Kuechler, B. Belitz, U. Trautmann, A. Schmidt, B. Schulze, L. Rodriguez, et al. New Immortalized Cell Lines of Patients With Small Supernumerary Marker Chromosome: Towards the Establishment of a Cell Bank J. Histochem. Cytochem., June 1, 2007; 55(6): 651 - 660. [Abstract] [Full Text] [PDF]

				U. Muller, G. Ernst, C. Melle, R. Guthke, and F. von Eggeling Convergence of the proteomic pattern in cancer Bioinformatics, June 1, 2006; 22(11): 1293 - 1296. [Abstract] [Full Text] [PDF]

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2005