|Publication Type||Journal Article|
|Year of Publication||2006|
|Authors||Tholouli, E, Hoyland, JA, Di Vizio, D, O'connell, F, Macdermott, SA, Twomey, D, Levenson, R, Yin, JALiu, Golub, TR, Loda, M, Byers, R|
|Journal||Biochem Biophys Res Commun|
|Date Published||2006 Sep 22|
|Keywords||Biopsy, Fixatives, Formaldehyde, Gene Expression Profiling, Humans, In Situ Hybridization, Oligonucleotide Probes, Quantum Dots, RNA, Messenger|
Gene expression mapping using microarray analysis has identified useful gene signatures for predicting outcome. However, little of this has been translated into clinically effective diagnostic tools as microarrays require high quality fresh-frozen tissue samples. We describe a methodology of multiplexed in situ hybridization (ISH) using a novel combination of quantum dot (QD)-labeled oligonucleotide probes and spectral imaging analysis in routinely processed, formalin-fixed paraffin embedded human biopsies. The conditions for QD-ISH were optimized using a poly d(T) oligonucleotide in decalcified bone marrow samples. Single and multiplex QD-ISH was performed in samples with acute leukemia and follicular lymphoma using oligonucleotide probes for myeloperoxidase, bcl-2, survivin, and XIAP. Spectral imaging was used for post hybridization tissue analysis, enabling separation of spatially colocalized signals. The method allows quantitative characterization of multiple gene expression using non-bleaching fluorochromes. This is expected to facilitate multiplex in situ transcript detection in routinely processed human clinical tissue.
|Alternate Journal||Biochem. Biophys. Res. Commun.|