Debarre, Delphine (2006) Third-harmonic generation microscopy for biological imaging. PhD thesis LOB, EP - LOB Laboratoire d'Optique et Biosciences, EP/X p.242.
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Alternative Locations: http://www.imprimerie.polytechnique.fr/Theses/Files/Debarre.pdf
Abstract
This work aimed at developing third-harmonic generation (THG) microscopy for biology. THG microscopy is a recent technique allowing the visualization of unstained cells and tissues with sub-micrometer resolution. Its application to biological imaging was until now limited by the lack of studies of contrast mechanisms inside samples, endogenous sources of signal in biology, and induced phototoxicity. The work presented here mainly addresses these three issues.
Firstly, we studied theoretically and experimentally the influence of sample structure and excitation beam focusing on THG signal. We showed that THG imaging acts on the sample as a bandpass filter for spatial frequencies and that adjusting the focusing of the excitation beam can be used to modulate the signal from structures different shapes within a complex sample.
Secondly, we characterized optical properties of several biological liquids, showing that lipid bodies embedded in an aqueous environment should provide a strong intracellular source of contrast. We then demonstrated that those structures can indeed be tracked and quantified in a variety of unstained tissues using THG microscopy.
Finally, we applied THG microscopy to the visualization of early development of live unstained Drosophila embryos. On this model, we studied the mechanisms of induced phototoxicity and demonstrated that THG microscopy can provide a complete 3D description of morphogenetic movements in wild-type and mutant embryos without perturbing embryo development.
| Item Type: | PhD Thesis (PhD) |
|---|---|
| Thesis Supervisor: | Beaurepaire, Emmanuel |
| Date: | September 2006 |
| Board of examiners: | Claude, Boccara and Jerome, Mertz and Jean-Paul, Thiery and Yaron, Silberberg and Jean-Louis, Martin |
| Ecole Doctorale: | ED 447 ECOLE DOCTORALE DE L'ECOLE POLYTECHNIQUE |
| Discipline: | LOB |
| Collection (Fonds): | EP/X |
| Institution: | EP/X |
| Department: | EP - LOB Laboratoire d'Optique et Biosciences |
| Subjects: | 3. Physics, Optics |
| Uncontrolled Keywords: | THG microscopy, Third-harmonic generation, Multiphoton microscopy, Tissue imaging, Lipid droplets, Phototoxicity, Drosophila melanogaster, microscopie THG, Génération de troisième harmonique, Microscopie multiphotonique, Imagerie des tissus, Gouttelettes lipidiques, Phototoxicité, Drosophila melanogaster |
Table of content
I-Introduction
II-Facteurs géométriques en imagerie THG
III-Sources de signal en microscopie THG
IV-Imagerie THG de l'embryon de drosophile
| ID Code: | 1978 |
|---|---|
| Deposited By: | Laurence Vidament |
| Deposited On: | 31 October 2006 |
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