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Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses
Thesis (PhD)--Stellenbosch University, 2024.
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| Format: | Thesis |
| Language: | English |
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Stellenbosch : Stellenbosch University
2025
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| _version_ | 1867613920370360320 |
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| access_status_str | Open Access |
| author | Badrodien, Imraan |
| author2 | Bosman, Gurthwin W.
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| author_browse | Badrodien, Imraan Bosman, Gurthwin W. |
| author_facet | Bosman, Gurthwin W.
Badrodien, Imraan |
| author_sort | Badrodien, Imraan |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (PhD)--Stellenbosch University, 2024. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/131565 |
| institution | Stellenbosch University (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:43:48.768Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/131565 Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses Badrodien, Imraan Bosman, Gurthwin W. Neethling, Pieter H. Stellenbosch University. Faculty of Science. Dept. of Physics. Imaging systems in biology Photonic crystals Nonlinear optics Two photon excitation fluorescence Light sheet fluorescence microscopy Multiphoton excitation microscopy UCTD Thesis (PhD)--Stellenbosch University, 2024. Badrodien, Imraan. 2024. Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses. Unpublished doctoral dissertation. Stellenbosch : Stellenbosch University [online]. Available: https://scholar.sun.ac.za/handle/10019.1/131565 ENGLISH ABSTRACT: Biological imaging has been transformed by the use of light-sheet fluorescence microscopy, which enables faster three-dimensional imaging with lower photobleaching and photo-toxicity compared to traditional microscopy. Nonlinear optical processes, specifically two-photon excitation fluorescence, have further improved imaging capabilities by enabling deeper penetration depths and reduced out-of-focus fluorescence. However, t he h igh p eak i ntensities required for nonlinear responses can lead to photodamage, making it imperative to maintain incident radiation below the damage threshold of biological samples when implementing nonlinear microscopy techniques. Recent advancements in all-normal-dispersion photonic crystal fibres (ANDi-PCF) have enabled the generation of highly stable broadband supercontinuum pulses that can be temporally compressed. While the multiphoton intrapulse interference phase scanning technique has been the popular choice for pulse characterisation, an alternative technique called i2PIE has been proposed and developed to reconstruct the spectral phase of a pulse. This dissertation reports the use of broadband supercontinuum pulses generated in an ANDi-PCF, compressed using the i2PIE phase measurement technique, in two-photon light-sheet microscopy. The performance is compared with other pulse compression techniques, and the application of this new method in imaging biological samples is demonstrated. The results indicate that the i2PIE technique enhanced the imaging capabilities of the two-photon light-sheet microscope, producing increased contrast and signal-to-noise ratios across all images compared to traditional compression techniques such as MIIPS or compression employing only the use of chirped mirrors. This demonstrates the potential of the i2PIE spectral phase measurement technique to enhance the imaging capabilities of two photon light-sheet fluorescence microscopy. AFRIKAANSE OPSOMMING: Biologiese beeldvorming is getransformeer deur die gebruik van ligblad fluo- ressensiemikroskopie, wat vinniger driedimensionele afbeelding moontlik maak met laer fotobleiking en fototoksisiteit in vergelyking met tradisionele mikro- skopie. Nie-lineêre optiese prosesse, spesifiek t wee-foton o pwekking fluores- sensie, het verder verbeterings in die afbeeldingsvermoëns moontlik gemaak deur dieper penetrasiedieptes en uitfokus fluoresensie. D ie h oë piekintensi- teite wat egter vereis word in nie-lineêre reaksies kan tot fotoskade lei, wat dit noodsaaklik maak om invallende bestraling onder die skadedrempel van bi- ologiese monsters te handhaaf wanneer nie-lineêre mikroskopietegnieke geim- plementeer word. Onlangse vooruitgang in alle-normale-dispersie fotoniese kristalvesels (ANDi-PCF) het die generasie van stabiele breëband superkonti- nuumpulse moontlik gemaak wat in tyd saamgepers kan word. Die multifoton intrapuls interferensie fase skandering tegniek was die gewilde keuse vir puls karakterisering. ’n Alternatiewe tegniek, i2PIE, was voorgestel en ontwikkel om die spektrale fase van ’n puls te rekonstrueer. Hierdie proefskrif rapporteer die gebruik van breëband superkontinuum pulse gegenereer in ’n ANDi-PCF, saamgepers met die i2PIE fasemetingstegniek, in twee-foton ligblad mikrosko- pie. Die verbeteringe word vergelyk met ander pulskompressietegnieke en die toepassing van hierdie nuwe metode in die beeldvorming van biologiese mon- sters word gedemonstreer. Die resultate dui aan dat die i2PIE-tegniek die beeldvermoë van die twee-foton ligbladmikroskoop verbeter het in vergelyking met tradisionele kompressietegnieke, in terme van verhoogte kontras en sein to geraas verhoudings oor all beelde. Die resultate beklemtoon die potensiaal van die i2PIE-spektrale fasemeting tegniek om die afbeeldvermoë van twee-foton ligblad fluoresensiemikroskopie te verbeter. Doctoral 2025-01-27T09:06:36Z 2025-01-27T09:06:36Z 2024-12 Thesis https://scholar.sun.ac.za/handle/10019.1/131565 en Stellenbosch University xvi, 104 pages : illustrations application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Imaging systems in biology Photonic crystals Nonlinear optics Two photon excitation fluorescence Light sheet fluorescence microscopy Multiphoton excitation microscopy UCTD Badrodien, Imraan Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title | Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title_full | Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title_fullStr | Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title_full_unstemmed | Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title_short | Two-photon light-sheet fluorescence microscopy with compressed supercontinuum pulses |
| title_sort | two photon light sheet fluorescence microscopy with compressed supercontinuum pulses |
| topic | Imaging systems in biology Photonic crystals Nonlinear optics Two photon excitation fluorescence Light sheet fluorescence microscopy Multiphoton excitation microscopy UCTD |
| url | https://scholar.sun.ac.za/handle/10019.1/131565 |
| work_keys_str_mv | AT badrodienimraan twophotonlightsheetfluorescencemicroscopywithcompressedsupercontinuumpulses |