Title High numerical aperture imaging allows chirality measurement in individual collagen fibrils using polarization second harmonic generation microscopy /
Authors Harvey, MacAulay ; Cisek, Richard ; Alizadeh, Mehdi ; Barzda, Virginijus ; Kreplak, Laurent ; Tokarz, Danielle
DOI 10.1515/nanoph-2023-0177
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Is Part of Nanophotonics.. Berlin : De Gruyter. 2023, vol. 12, iss. 11, p. 2061-2071.. ISSN 2192-8606. eISSN 2192-8614
Keywords [eng] atomic force microscopy ; nano-biophotonics ; nanofibers ; nanoscale structures ; nonlinear optical microscopy ; polarization-resolved imaging
Abstract [eng] Second harmonic generation (SHG) microscopy is a commonly used technique to study the organization of collagen within tissues. However, individual collagen fibrils, which have diameters much smaller than the resolution of most optical systems, have not been extensively investigated. Here we probe the structure of individual collagen fibrils using polarization-resolved SHG (PSHG) microscopy and atomic force microscopy. We find that longitudinally polarized light occurring at the edge of a focal volume of a high numerical aperture microscope objective illuminated with linearly polarized light creates a measurable variation in PSHG signal along the axis orthogonal to an individual collagen fibril. By comparing numerical simulations to experimental data, we are able to estimate parameters related to the structure and chirality of the collagen fibril without tilting the sample out of the image plane, or cutting tissue at different angles, enabling chirality measurements on individual nanostructures to be performed in standard PSHG microscopes. The results presented here are expected to lead to a better understanding of PSHG results from both collagen fibrils and collagenous tissues. Further, the technique presented can be applied to other chiral nanoscale structures such as microtubules, nanowires, and nanoribbons.
Published Berlin : De Gruyter
Type Journal article
Language English
Publication date 2023
CC license CC license description