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

Richard J. Byers, Dolores Di Vizio, Fionnuala O'Connell, Eleni Tholouli, Richard M Levenson, Kirk Gossard, David Twomey, Yu Yang, Elisa Benedettini, Joshua Rose, Keith L. Ligon, Stephen P. Finn, Todd R. Golub, Massimo Loda

Research output: Contribution to journalArticlepeer-review

47 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)20-29
Number of pages10
JournalJournal of Molecular Diagnostics
Issue number1
StatePublished - Feb 2007
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology


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