Imaging of multiple mRNA targets using quantum dot based in situ hybridization and spectral deconvolution in clinical biopsies

Eleni Tholouli, Judith A. Hoyland, Dolores Di Vizio, Fionnuala O'Connell, Sarah A. MacDermott, David Twomey, Richard M Levenson, John A Liu Yin, Todd R. Golub, Massimo Loda, Richard Byers

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Gene expression mapping using microarray analysis has identified useful gene signatures for predicting outcome. However, little of this has been translated into clinically effective diagnostic tools as microarrays require high quality fresh-frozen tissue samples. We describe a methodology of multiplexed in situ hybridization (ISH) using a novel combination of quantum dot (QD)-labeled oligonucleotide probes and spectral imaging analysis in routinely processed, formalin-fixed paraffin embedded human biopsies. The conditions for QD-ISH were optimized using a poly d(T) oligonucleotide in decalcified bone marrow samples. Single and multiplex QD-ISH was performed in samples with acute leukemia and follicular lymphoma using oligonucleotide probes for myeloperoxidase, bcl-2, survivin, and XIAP. Spectral imaging was used for post hybridization tissue analysis, enabling separation of spatially colocalized signals. The method allows quantitative characterization of multiple gene expression using non-bleaching fluorochromes. This is expected to facilitate multiplex in situ transcript detection in routinely processed human clinical tissue.

Original languageEnglish (US)
Pages (from-to)628-636
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume348
Issue number2
DOIs
StatePublished - Sep 22 2006
Externally publishedYes

Fingerprint

Quantum Dots
Biopsy
Deconvolution
Semiconductor quantum dots
In Situ Hybridization
Oligonucleotide Probes
Tissue
Microarrays
Imaging techniques
Gene expression
Messenger RNA
Poly T
Gene Expression
Follicular Lymphoma
Chromosome Mapping
Microarray Analysis
Fluorescent Dyes
Oligonucleotides
Paraffin
Formaldehyde

Keywords

  • Bone marrow biopsies
  • In situ hybridization
  • Messenger RNA detection
  • Multiplex in situ hybridization
  • Quantum dots
  • Spectral deconvolution

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Imaging of multiple mRNA targets using quantum dot based in situ hybridization and spectral deconvolution in clinical biopsies. / Tholouli, Eleni; Hoyland, Judith A.; Di Vizio, Dolores; O'Connell, Fionnuala; MacDermott, Sarah A.; Twomey, David; Levenson, Richard M; Yin, John A Liu; Golub, Todd R.; Loda, Massimo; Byers, Richard.

In: Biochemical and Biophysical Research Communications, Vol. 348, No. 2, 22.09.2006, p. 628-636.

Research output: Contribution to journalArticle

Tholouli, E, Hoyland, JA, Di Vizio, D, O'Connell, F, MacDermott, SA, Twomey, D, Levenson, RM, Yin, JAL, Golub, TR, Loda, M & Byers, R 2006, 'Imaging of multiple mRNA targets using quantum dot based in situ hybridization and spectral deconvolution in clinical biopsies', Biochemical and Biophysical Research Communications, vol. 348, no. 2, pp. 628-636. https://doi.org/10.1016/j.bbrc.2006.07.122
Tholouli, Eleni ; Hoyland, Judith A. ; Di Vizio, Dolores ; O'Connell, Fionnuala ; MacDermott, Sarah A. ; Twomey, David ; Levenson, Richard M ; Yin, John A Liu ; Golub, Todd R. ; Loda, Massimo ; Byers, Richard. / Imaging of multiple mRNA targets using quantum dot based in situ hybridization and spectral deconvolution in clinical biopsies. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 348, No. 2. pp. 628-636.
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