Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy

Jing Liu, Nur P. Damayanti, Il Hoon Cho, Yesim Polar, Sunil Badve, Joseph M.K. Irudayaraj

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Fluorescence-based single molecule techniques to interrogate gene expression in tissues present a very low signal-to-noise ratio due to the strong autofluorescence and other background signals from tissue sections. This report presents a background-free method using second-harmonic generation (SHG) nanocrystals as probes to quantify the messenger RNA (mRNA) of human epidermal growth receptor 2 (Her2) at single molecule resolution in specific phenotypes at single-cell resolution directly in tissues. Coherent SHG emission from individual barium titanium oxide (BTO) nanoprobes was demonstrated, allowing for a stable signal beyond the autofluorescence window. Her2 surface marker and Her2 mRNA were specifically labeled with BTO probes, and Her2 mRNA was quantified at single copy sensitivity in Her2 expressing phenotypes directly in cancer tissues. Our approach provides the first proof of concept of a cross-platform strategy to probe tissues at single-cell resolution in situ.

Original languageEnglish (US)
Article number096016
JournalJournal of Biomedical Optics
Volume20
Issue number9
DOIs
StatePublished - Sep 1 2015

Keywords

  • barium titanium oxide nanocrystals
  • mRNA quantification
  • second-harmonic generation
  • tissues

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics

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