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

J Mol Diagn
Authors
Keywords
Abstract

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.

Year of Publication
2007
Journal
J Mol Diagn
Volume
9
Issue
1
Pages
20-9
Date Published
2007 Feb
ISSN
1525-1578
URL
DOI
10.2353/jmoldx.2007.060119
PubMed ID
17251332
PubMed Central ID
PMC2248801
Links
Grant list
P01 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