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Journal of Biomedical Optics • Open Access

Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy
Author(s): Jing Liu; Nur P. Damayanti; Il-Hoon Cho; Yesim Polar; Sunil Badve; Joseph M. K. Irudayaraj

Paper 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.

Paper Details

Date Published: 25 September 2015
PDF: 7 pages
J. Biomed. Opt. 20(9) 096016 doi: 10.1117/1.JBO.20.9.096016
Published in: Journal of Biomedical Optics Volume 20, Issue 9
Show Author Affiliations
Jing Liu, Purdue Univ. (United States)
South Dakota School of Mines and Technology (United States)
Nur P. Damayanti, Purdue Univ. (United States)
Il-Hoon Cho, Purdue Univ. (United States)
Eulji Univ. (Korea, Republic of)
Yesim Polar, Indiana Univ. School of Medicine (United States)
Sunil Badve, Indiana Univ. School of Medicine (United States)
Joseph M. K. Irudayaraj, Purdue Univ. (United States)

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