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dc.contributor.advisorAhluwalia, Balpreet Singh
dc.contributor.authorJayakumar, Nikhil
dc.date.accessioned2024-01-24T12:07:43Z
dc.date.available2024-01-24T12:07:43Z
dc.date.issued2024-02-06
dc.description.abstractTo see structures well below 200 nm using visible light, super-resolution techniques have been developed. Most of these techniques rely on fluorescence labelling or use of artificial materials to go beyond the Abbe limit. Therefore, achieving super-resolution in the label-free regime is the problem that is being addressed in this thesis. In this work, high-index contrast dielectric waveguides are employed for microscopy. Typically, wide waveguides are employed for bio-imaging applications. High-index contrast wide waveguides support multiple optical modes to guide optical power along its length. Therefore, the first task was to use multiple modes to artificially induce intensity fluctuations that are compatible with fluorescence based super-resolution algorithms. This enabled the generation of super-resolved fluorescence images. The next task was to see if these multiple-modes can be used to illuminate unlabeled samples. By averaging several images within the integration time of the camera, a high-contrast label-free image was generated using these waveguides. Finally, to achieve super-resolution in label-free mode, photoluminescence in Si3N4 waveguides was used for near-field illumination of unlabeled samples. This helped realize a label-free incoherent imaging system, thus permitting the application of fluorescence-based super-resolution algorithms to achieve super-resolution in label-free mode.en_US
dc.description.doctoraltypeph.d.en_US
dc.description.popularabstractTwo major challenges in optical microscopy, since its invention, are poor contrast and diffraction-limited resolution. Well, what do they mean? Poor contrast means that you cannot discern your structure of interest very well from its background. The situation is analogous to a piece of glass inside a bowl of water. This piece of glass is almost invisible when seen from outside. This is exactly what happens when you try to image biological cells. They are like this ‘glass piece’ inside the bowl of water. So how can you see them? Maybe you can color them? Yes, exactly! You add certain dyes to your biological cells to make them visible. This is what is done in fluorescence microscopy. This field of research is quite mature in life sciences and is a common practice in labs across the world. Now, let us look at the second challenge. Let us analyze each word in ‘diffraction-limited resolution.’ Resolution means what is the closest distance you can bring two particles together, before they start appearing as one. For example, when you stare at the mountains from your homes, you do not see the vegetation or the leaves on the trees. You see a big mountain and that’s it. This is what resolution means, how much detail you can see. And the reason why the resolution is poor is due to a physical property called the diffraction of light. What this means is that light microscope, we cannot see structures or details below 200 nm. Luckily, fluorescence microscopy helps mitigate this problem. In fluorescence microscopy, additional information is obtained by playing with the fluorescent dye. For e.g., by making them blink, resolution below 200 nm can be achieved using certain computational techniques. However, cells like to remain in their natural environment, i.e., they do not want any artificial labelling to be done on them. Therefore, a big challenge is to image these cells with very high resolution, < 200 nm, without labeling them. This field is called label-free microscopy and has been gaining popularity in high resolution label-free microscopy ever since the development of the concept of negative lens. Though, several techniques have evolved over the last two decades, the field of label-free microscopy is yet to catch up like fluorescence microscopy due to experimental challenges. Therefore, in this thesis, label-free super-resolution microscopy is the problem that is being attacked. The idea is to apply computational techniques developed for super-resolution fluorescence algorithms in the label-free regime. However, directly applying these techniques is conceptually wrong. This thesis therefore, provides a solution to this problem: use tiny light sources for illumination of unlabeled samples. By realizing such an experimental setup, one can generate super-resolved two dimensional label-free images. The developed technique has been experimentally tested on nanobeads and biological specimens. In future, the concepts developed in this thesis can help researchers build three dimensional label-free super-resolved microscopes.en_US
dc.description.sponsorshipThis project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement No. 766181, project “DeLIVER”.en_US
dc.identifier.isbn978-82-8236-562-8 (trykt), 978-82-8236-563-5 (pdf)
dc.identifier.urihttps://hdl.handle.net/10037/32703
dc.language.isoengen_US
dc.publisherUiT Norges arktiske universiteten_US
dc.publisherUiT The Arctic University of Norwayen_US
dc.relation.haspart<p>Paper I: Jayakumar, N., Helle, Ø.I., Agarwal, K. & Ahluwalia, B.S. (2020). On-chip TIRF nanoscopy by applying Haar wavelet kernel analysis on intensity fluctuations induced by chip illumination. <i>Optics Express, 28</i>(24), 35454-35468. Also available in Munin at <a href=https://hdl.handle.net/10037/19930>https://hdl.handle.net/10037/19930</a>. <p>Paper II: Jayakumar, N., Dullo, F.T., Dubey, V., Ahmad, A., Ströhl, F., Cauzzo, J., … Ahluwalia, B.S. (2022). Multi-moded high-index contrast optical waveguide for super-contrast high-resolution label-free microscopy. <i>Nanophotonics, 11</i>(15), 3421-3436. Also available in Munin at <a href=https://hdl.handle.net/10037/26604>https://hdl.handle.net/10037/26604</a>. <p>Paper III: Jayakumar, N., Villegas-Hernández, L.E., Zhao, W., Mao, H., Dullo, F.T., Tinguley, J.C., … Ahluwalia, B.S. Label-free incoherent superresolution optical microscopy. (Manuscript). Also available on arXiv at <a href=https://doi.org/10.48550/arXiv.2301.03451>https://doi.org/10.48550/arXiv.2301.03451</a>.en_US
dc.relation.projectIDinfo:eu-repo/grantAgreement/EC/H2020/766181/EU/Super-resolution optical microscopy of nanosized pore dynamics in endothelial cells/DeLIVER/en_US
dc.rights.accessRightsopenAccessen_US
dc.rights.holderCopyright 2024 The Author(s)
dc.subject.courseIDDOKTOR-004
dc.subjectOpticsen_US
dc.subjectMicrscopyen_US
dc.subjectApplied Physicsen_US
dc.titleLabel-free super-resolution optical microscopyen_US
dc.typeDoctoral thesisen_US
dc.typeDoktorgradsavhandlingen_US


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