Proteohistography--direct analysis of tissue with high sensitivity and high spatial resolution using ProteinChip technology.
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. 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. 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, surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS), allows the laser to be steered 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 distribution proteohistologically. Results show that the spatial distribution of detectable proteins could be used as a Proteohistogram for a given tissue area. Consequently, this procedure can provide additional information to both a matrix-assisted laser desorption/ionization (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.
Ernst
Günther
G
Institut für Humangenetik und Anthropologie, CUCA, 07740 Jena, Germany.
Melle
Christian
C
Schimmel
Bettina
B
Bleul
Annett
A
von Eggeling
Ferdinand
F
D006528:Carcinoma, Hepatocellular / Q000737:chemistry; D003106:Colon / Q000737:chemistry; D003110:Colonic Neoplasms / Q000737:chemistry; D005753:Gastric Mucosa / Q000737:chemistry; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008113:Liver Neoplasms / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D020543:Proteome / Q000032:analysis*; D012680:Sensitivity and Specificity; D013270:Stomach / Q000737:chemistry; D013274:Stomach Neoplasms / Q000737:chemistry
D020543:Proteome