Volume 8, Issue 9 pp. 730-739
Full Article

Polarization second harmonic generation microscopy provides quantitative enhanced molecular specificity for tissue diagnostics

Rajesh Kumar

Corresponding Author

Rajesh Kumar

Department of Physics, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway

Corresponding author: e-mail: [email protected]

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Kirsten M. Grønhaug

Kirsten M. Grønhaug

Levanger Hospital, Norway

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Elisabeth I. Romijn

Elisabeth I. Romijn

Department of Physics, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway

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Andreas Finnøy

Andreas Finnøy

Department of Physics, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway

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Catharina L. Davies

Catharina L. Davies

Department of Physics, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway

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Jon O. Drogset

Jon O. Drogset

Trondheim University Hospital, Norway

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Magnus B. Lilledahl

Magnus B. Lilledahl

Department of Physics, Norwegian University of Science and Technology (NTNU), N-7491 Trondheim, Norway

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First published: 03 November 2014
Citations: 40

Abstract

Due to specific structural organization at the molecular level, several biomolecules (e.g., collagen, myosin etc.) which are strong generators of second harmonic generation (SHG) signals, exhibit unique responses depending on the polarization of the excitation light. By using the polarization second harmonic generation (p-SHG) technique, the values of the second order susceptibility components can be used to differentiate the types of molecule, which cannot be done by the use of a standard SHG intensity image. In this report we discuss how to implement p-SHG on a commercial multiphoton microscope and overcome potential artifacts in susceptibility (χ) image. Furthermore we explore the potential of p-SHG microscopy by applying the technique to different types of tissue in order to determine corresponding reference values of the ratio of second-order χ tensor elements. These values may be used as a bio-marker to detect any structural alterations in pathological tissue for diagnostic purposes.

figure

The SHG intensity image (red) in (a) shows the distribution of collagen fibers in ovary tissue but cannot determine the type of collagen fiber. However, the histogram distribution (b) for the values of the χ tensor element ratio can be used to quantitatively identify the types of collagen fibers.

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