25 September 2015 Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy
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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
© 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
Jing Liu, Jing Liu, Nur P. Damayanti, Nur P. Damayanti, Il-Hoon Cho, Il-Hoon Cho, Yesim Polar, Yesim Polar, Sunil Badve, Sunil Badve, Joseph M. K. Irudayaraj, Joseph M. K. Irudayaraj, } "Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy," Journal of Biomedical Optics 20(9), 096016 (25 September 2015). https://doi.org/10.1117/1.JBO.20.9.096016 . Submission:
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