Research Papers: Imaging

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

[+] Author Affiliations
Jing Liu

Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United States

South Dakota School of Mines and Technology, Nanoscience and Nanoengineering, Rapid City, South Dakota 57701, United States

Nur P. Damayanti, Joseph M. K. Irudayaraj

Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United States

Il-Hoon Cho

Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United States

Eulji University, College of Health Science, Department of Biomedical Laboratory Science, Seongnam 461–713, Republic of Korea

Yesim Polar, Sunil Badve

Indiana University School of Medicine, Department of Pathology and Laboratory Medicine, Indianapolis, Indiana 46202, United States

J. Biomed. Opt. 20(9), 096016 (Sep 25, 2015). doi:10.1117/1.JBO.20.9.096016
History: Received April 27, 2015; Accepted August 26, 2015
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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.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Jing Liu ; Nur P. Damayanti ; Il-Hoon Cho ; Yesim Polar ; Sunil Badve, et al.
"Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy", J. Biomed. Opt. 20(9), 096016 (Sep 25, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.9.096016


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