Abstract
We provide evidence that the chirality of collagen can give rise to strong second-harmonic generation circular dichroism (SHG-CD) responses in nonlinear microscopy. Although chirality is an intrinsic structural property of collagen, most of the previous studies ignore that property. We demonstrate chiral imaging of individual collagen fibers by using a laser scanning microscope and type-I collagen from pig ligaments. 100% contrast level of SHG-CD is achieved with sub-micrometer spatial resolution. As a new contrast mechanism for imaging chiral structures in bio-tissues, this technique provides information about collagen morphology and three-dimensional orientation of collagen molecules.
Highlights
Chirality, lack of mirror symmetry, is an important and fundamental structural property of matter, and can be extensively found in various biological molecules, such as proteins, nucleic acids, and saccharides
We show that collagens can give rise to strong second-harmonic generation circular dichroism (SHG-circular dichroism (CD)) responses, with 100% contrast level, under a polarized nonlinear optical microscope
Similar to conventional CD, the contrast of Second-harmonic generation (SHG)-CD response is defined as the difference between ILCP(2ω) and IRCP(2ω), divided by their average, given as [35]: I SHG −CD
Summary
Lack of mirror symmetry, is an important and fundamental structural property of matter, and can be extensively found in various biological molecules, such as proteins, nucleic acids, and saccharides. Y. Chen, “Reconstruction of complementary images in second harmonic generation microscopy,” Opt. Express 14(11), 4727–4735 (2006). “Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy,” Opt. Express 15(19), 12286–12295 (2007). “Quantification of the second-order nonlinear susceptibility of collagen I using a laser scanning microscope,” J.
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