期刊
OPTICA
卷 8, 期 5, 页码 674-685出版社
OPTICAL SOC AMER
DOI: 10.1364/OPTICA.411325
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资金
- European Research Council [638258]
- Engineering and Physical Sciences Research Council [EP/N509747/1, EP/T020997/1]
- Leverhulme Trust [RPG-2018-251]
- Wellcome Trust [100638/Z/12/Z]
- NIHR Southampton Biomedical Research Centre
- Wellcome Trust [100638/Z/12/Z] Funding Source: Wellcome Trust
- EPSRC [EP/T020997/1] Funding Source: UKRI
The study utilizes photonic nanojet phenomenon to achieve super-resolution second-harmonic generation (SR-SHG) imaging, improving resolution by approximately 2.3 times compared to traditional methods. This technique allows observation of changes in SHG-active structures such as collagen at the nanoscale, offering a potential new tool for biological research and drug discovery.
Superresolution (SR) optical microscopy has allowed the investigation of many biological structures below the diffraction limit; however, most of the techniques are hampered by the need for fluorescent labels. Nonlinear label-free techniques such as second-harmonic generation (SHG) provide structurally specific contrast without the addition of exogenous labels, allowing observation of unperturbed biological systems. We use the photonic nanojet (PNJ) phenomena to achieve SR-SHG. A resolution of similar to lambda/6 with respect to the fundamental wavelength, that is, a similar to 2.3-fold improvement over conventional or diffraction-limited SHG under the same imaging conditions is achieved. Crucially we find that the polarization properties of excitation are maintained in a PNJ. This is observed in experiment and simulations. This may have widespread implications to increase sensitivity by detection of polarization-resolved SHG by observing anisotropy in signals. These new, to the best of our knowledge, findings allowed us to visualize biological SHG-active structures such as collagen at an unprecedented and previously unresolvable spatial scale. Moreover, we demonstrate that the use of an array of self-assembled high-index spheres overcomes the issue of a limited field of view for such a method, allowing PNJ-assisted SR-SHG to be used over a large area. Dysregulation of collagen at the nanoscale occurs in many diseases and is an underlying cause in diseases such as lung fibrosis. Here we demonstrate that pSR-SHG allows unprecedented observation of changes at the nanoscale that are invisible by conventional diffraction-limited SHG imaging. The ability to nondestructively image SHG-active biological structures without labels at the nanoscale with a relatively simple optical method heralds the promise of a new tool to understand biological phenomena and drive drug discovery. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License.
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