Title | Semiautomated multiplexed quantum dot-based in situ hybridization and spectral deconvolution. |
Publication Type | Journal Article |
Year of Publication | 2007 |
Authors | Byers 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 |
Journal | J Mol Diagn |
Volume | 9 |
Issue | 1 |
Pagination | 20-9 |
Date Published | 2007 Feb |
ISSN | 1525-1578 |
Keywords | Animals, 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 |
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. |
DOI | 10.2353/jmoldx.2007.060119 |
Alternate Journal | J Mol Diagn |
PubMed ID | 17251332 |
PubMed Central ID | PMC2248801 |
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 |
Related Faculty:
Massimo Loda, M.D.