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 journalArticle

5 Citations (Scopus)

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
Article number096016
JournalJournal of Biomedical Optics
Volume20
Issue number9
DOIs
StatePublished - Sep 1 2015

Fingerprint

Harmonic generation
harmonic generations
Microscopic examination
ribonucleic acids
Screening
screening
cancer
Tissue
microscopy
resolution cell
barium oxides
cells
phenotype
Titanium oxides
Barium
titanium oxides
Messenger RNA
probes
Nanoprobes
Molecules

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

Cite this

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

In: Journal of Biomedical Optics, Vol. 20, No. 9, 096016, 01.09.2015.

Research output: Contribution to journalArticle

@article{363b3b76fdff4203ab35d7d23222fe16,
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.",
keywords = "barium titanium oxide nanocrystals, mRNA quantification, second-harmonic generation, tissues",
author = "Jing Liu and Damayanti, {Nur P.} and Cho, {Il Hoon} and Yesim Polar and Sunil Badve and Irudayaraj, {Joseph M K}",
year = "2015",
month = "9",
day = "1",
doi = "10.1117/1.JBO.20.9.096016",
language = "English",
volume = "20",
journal = "Journal of Biomedical Optics",
issn = "1083-3668",
publisher = "SPIE",
number = "9",

}

TY - JOUR

T1 - Single-cell screening and quantification of transcripts in cancer tissues by second-harmonic generation microscopy

AU - Liu, Jing

AU - Damayanti, Nur P.

AU - Cho, Il Hoon

AU - Polar, Yesim

AU - Badve, Sunil

AU - Irudayaraj, Joseph M K

PY - 2015/9/1

Y1 - 2015/9/1

N2 - 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.

AB - 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.

KW - barium titanium oxide nanocrystals

KW - mRNA quantification

KW - second-harmonic generation

KW - tissues

UR - http://www.scopus.com/inward/record.url?scp=84942585457&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84942585457&partnerID=8YFLogxK

U2 - 10.1117/1.JBO.20.9.096016

DO - 10.1117/1.JBO.20.9.096016

M3 - Article

C2 - 26405822

AN - SCOPUS:84942585457

VL - 20

JO - Journal of Biomedical Optics

JF - Journal of Biomedical Optics

SN - 1083-3668

IS - 9

M1 - 096016

ER -