Scientific Publications

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

Publication TypeJournal Article
AuthorsByers, RJ, Di Vizio D., O'connell F., Tholouli E., Levenson RM, Gossage K., Twomey D., Yang Y., Benedettini E., Rose J., Ligon KL, Finn SP, Golub T. R., and Loda M.
AbstractGene 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.
Year of Publication2007
JournalThe Journal of molecular diagnostics : JMD
Date Published (YYYY/MM/DD)2007/02/01
ISSN Number1525-1578