Molecular high throughput analyses and automated detection systems

Abstract

High throughput equipment, like tissue arrayer and autostainer, has meanwhile been integrated in pathology and research on a daily-use basis and provides comparability, reproducibility and celerity of processing. Since molecular investigations possessing high throughput characteristics (transcription arrays and 2-D-electrophoresis) can be combined with this equipment, effi cient strategies to fi nd disease-relevant molecules can be developed. In this study transcription array data derived from human lungs, tumor and tumor-free specimens, was evaluated to disclose strong gene regulations. Candidate molecules, one of which was haptoglobin, were found and studied intensively employing high throughput immunohistochemistry. For evaluation of signals from immunohistochemistry, an automated stand-alone tissue microarray detection system (“Spot Browser”) was used to analyze stained tissue microarrays. We describe diff erent technical and molecular high throughput tools whose combination represents an effi cient strategy for the identifi cation of most important candidate molecules with high importance in the fi eld of lung diseases including cancer, infl ammation, allergy, and infection. All tissue conservation was further based on the formalin-, ethanoland xylolfree HOPE-fi xation which provides advantageous molecular conservation due to its gentle fi xation mechanism. Th is technique for paraffi n-embedded material maintains morphology equal to formalin and avoids protein crosslinking and degradation of RNA and DNA in a long-term manner; moreover, all molecular-biologic and biochemical methods are compatible. In conclusion, HOPE-fi xed tissues processed with high throughput equipment and analyzed using molecular high throughput techniques provide a powerful combination as the basis for multimethodical investigations of archived tissue specimens and therefore for the fast detection of disease-associated molecules.