Semiautomated multiplexed quantum dot-based in situ hybridization and spectral deconvolution. Academic Article uri icon

Overview

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.

publication date

  • February 1, 2007

Research

keywords

  • Cell Lineage
  • Gene Expression Profiling
  • In Situ Hybridization
  • Molecular Diagnostic Techniques
  • Quantum Dots

Identity

PubMed Central ID

  • PMC2248801

Scopus Document Identifier

  • 33947507524

PubMed ID

  • 17251332

Additional Document Info

volume

  • 9

issue

  • 1