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760 collagen spectral stack .tif (1.38 MB)
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780 collagen spectral stack.tif (1.38 MB)
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840 collagen spectral stack.tif (1.38 MB)
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940 collagen spectral stack.tif (1.38 MB)
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fibroblast collagen 860 420.tif (769 kB)
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fibroblast collagen 920 460.tif (769 kB)
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Fibroblast collagen SHG intensity using narrow band filters.xlsx (23.38 kB)
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Data for paper "A new mode of contrast in biological second harmonic generation microscopy"

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posted on 2017-10-06, 09:20 authored by Nicola GreenNicola Green, Robin M Delaine-Smitth, Hannah J Askew, Robert Byers, Gwendolen ReillyGwendolen Reilly, Stephen MatcherStephen Matcher

Enhanced image contrast in biological second harmonic imaging microscopy (SHIM) has previously been reported via quantitative assessments of forward- to epi-generated signal intensity ratio and by polarization analysis. Here we demonstrate a new form of contrast: the material-specific, wavelength-dependence of epi-generated second harmonic generation (SHG) excitation efficiency, and discriminate collagen and myosin by ratiometric epi-generated SHG images at 920 nm and 860 nm.

Collagen shows increased SHG intensity at 920 nm, while little difference is detected between the two for myosin; allowing SHIM to characterize different SHG-generating components within a complex biological sample.


Scientific Reports 7(1) Article number:13331 (2017)

DOI: 10.1038/s41598-017-13752-y

Funding

EPSRC grants EP/I018328/1 & EP/K009699/1 and the University of Sheffield Women Academic Returners Program.

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    Department of Materials Science and Engineering

    Categories

    Keywords

    second harmonic generationtwo photon imagingcollagenmyosinwavelength dependencyBiomaterialsNonlinear Optics and SpectroscopyBiophysics

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