iSERS microscopy guided by wide field immunofluorescence: analysis of HER2 expression on normal and breast cancer FFPE tissue sections.
Surface-enhanced Raman scattering (SERS) microscopy is an emerging imaging technique for tissue-based cancer diagnostics. Specifically, immuno-SERS (iSERS) microscopy employs antibodies labelled by molecularly functionalized noble metal colloids for antigen localization on tissue specimen. Spectrally resolved iSERS acquisition schemes are typically rather time-consuming when large tissue areas must be scanned. Here, we demonstrate the application of iSERS imaging guided by wide field immunofluorescence (IF) for localization of the human epidermal growth factor receptor 2 (HER2) on breast tissue sections. The addition of unlabelled anti-HER2 primary antibodies to the tissue is followed by the incubation with secondary antibodies labelled with both Alexa-647 (for IF) and hydrophilically stabilized gold nanostars coated with aromatic thiols (for iSERS). False-color iSERS images clearly reveal the different HER2 expression levels on normal and breast cancer tissue, respectively. A series of negative controls confirms that the binding specificity of the secondary antibody is maintained after conjugation to the SERS nanoparticles.
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Xin-Ping
XP
Wang
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Yuying
Y
Zhang
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Matthias
M
König
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Evanthia
E
Papadopoulou
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Bernd
B
Walkenfort
Department of Gynecology and Obstetrics, University Hospital Essen, University of Duisburg-Essen, Hufelandstrasse 55, D-45122 Essen, Germany.
Sabine
S
Kasimir-Bauer
Institute of Pathology, University Hospital Essen, Hufelandstrasse 55, D-45122 Essen, Germany.
Agnes
A
Bankfalvi
Physical Chemistry I, Department of Chemistry and Center for Nanointegration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätsstr. 5, Essen 45141, Germany. sebastian.schluecker@uni-due.de.
Sebastian
S
Schlücker
D001940:Breast / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006046:Gold; D006801:Humans; D053768:Metal Nanoparticles; D008853:Microscopy; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D013059:Spectrum Analysis, Raman
D006046:Gold; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human