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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Biomed+Opt
2015 ; 20
(9
): 096016
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Single-cell screening and quantification of transcripts in cancer tissues by
second-harmonic generation microscopy
#MMPMID26405822
Liu J
; Damayanti NP
; Cho IH
; Polar Y
; Badve S
; Irudayaraj JM
J Biomed Opt
2015[Sep]; 20
(9
): 096016
PMID26405822
show ga
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.