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J Mol Diagn DOI:10.2353/jmoldx.2007.060119

Semiautomated multiplexed quantum dot-based in situ hybridization and spectral deconvolution.

Publication TypeJournal Article
Year of Publication2007
AuthorsByers, RJ, Di Vizio, D, O'connell, F, Tholouli, E, Levenson, RM, Gossage, K, Gossard, K, Twomey, D, Yang, Y, Benedettini, E, Rose, J, Ligon, KL, Finn, SP, Golub, TR, Loda, M
JournalJ Mol Diagn
Date Published2007 Feb
KeywordsAnimals, Cell Lineage, DNA, Complementary, Gene Expression Profiling, Humans, Image Processing, Computer-Assisted, Immunohistochemistry, In Situ Hybridization, Mice, Molecular Diagnostic Techniques, Oligonucleotide Probes, Quantum Dots, Sensitivity and Specificity, Tumor Cells, Cultured

Gene expression profiling has identified several potentially useful gene signatures for predicting outcome or for selecting targeted therapy. However, these signatures have been developed in fresh or frozen tissue, and there is a need to apply them to routinely processed samples. Here, we demonstrate the feasibility of a potentially high-throughput methodology combining automated in situ hybridization with quantum dot-labeled oligonucleotide probes followed by spectral imaging for the detection and subsequent deconvolution of multiple signals. This method is semiautomated and quantitative and can be applied to formalin-fixed, paraffin-embedded tissues. We have combined dual in situ hybridization with immunohistochemistry, enabling simultaneous measurement of gene expression and cell lineage determination. The technique achieves levels of sensitivity and specificity sufficient for the potential application of known expression signatures to biopsy specimens in a semiquantitative way, and the semiautomated nature of the method enables application to high-throughput studies.


Alternate JournalJ Mol Diagn
PubMed ID17251332
PubMed Central IDPMC2248801
Grant ListP01 CA089021 / CA / NCI NIH HHS / United States
P50 CA090381 / CA / NCI NIH HHS / United States
2P01 CA089021 / CA / NCI NIH HHS / United States
5P50CA90381 / CA / NCI NIH HHS / United States