title: Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy.
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.
authors.affiliation: Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United StatesdSouth Dakota School of Mines and Technology, Nanoscience and Nanoengineering, Rapid City, So.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United States.
authors.forename: Nur P
authors.initials: NP
authors.lastname: Damayanti
authors.affiliation: Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United StatesbEulji University, College of Health Science, Department of Biomedical Laboratory Science, Se.
authors.forename: Il-Hoon
authors.initials: IH
authors.lastname: Cho
authors.affiliation: Indiana University School of Medicine, Department of Pathology and Laboratory Medicine, Indianapolis, Indiana 46202, United States.
authors.forename: Yesim
authors.initials: Y
authors.lastname: Polar
authors.affiliation: Indiana University School of Medicine, Department of Pathology and Laboratory Medicine, Indianapolis, Indiana 46202, United States.
authors.forename: Sunil
authors.initials: S
authors.lastname: Badve
authors.affiliation: Purdue University, Bindley Bioscience Center and Birck Nanotechnology Center, Agriculture and Biological Engineering, West Lafayette, Indiana 47907, United States.
authors.forename: Joseph M K
authors.initials: JM
authors.lastname: Irudayaraj
mesh_terms: D017609:Barium Compounds; D001943:Breast Neoplasms / Q000737:chemistry / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D053768:Metal Nanoparticles; D008853:Microscopy / Q000379:methods*; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D059629:Signal-To-Noise Ratio; D059010:Single-Cell Analysis / Q000379:methods*; D014025:Titanium; D059467:Transcriptome / Q000235:genetics*
chemical_list: D017609:Barium Compounds; D012333:RNA, Messenger; C024547:barium titanate(IV); D014025:Titanium