Unravelling motor protein organization on lysosomal membranes with super-resolution microscopy
This thesis first develops new methods for high-throughput and multi-color super-resolution microscopy (Chapters 2 and 3). Subsequently, I apply these methods to study the organization of motor proteins on the lysosome membrane inside cells with the purpose of determining how intracellular transport...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2018 |
| País: | España |
| Institución: | CBUC, CESCA |
| Repositorio: | TDR. Tesis Doctorales en Red |
| OAI Identifier: | oai:www.tdx.cat:10803/565415 |
| Acceso en línea: | http://hdl.handle.net/10803/565415 https://dx.doi.org/10.5821/dissertation-2117-117629 |
| Access Level: | acceso abierto |
| Palabra clave: | Àrees temàtiques de la UPC::Física 53 535 57 |
| Sumario: | This thesis first develops new methods for high-throughput and multi-color super-resolution microscopy (Chapters 2 and 3). Subsequently, I apply these methods to study the organization of motor proteins on the lysosome membrane inside cells with the purpose of determining how intracellular transport can be regulated via motor-protein organization (Chapter 4). Chapter 1 is an Introduction to the state of the art for our knowledge in microtubule-based intracellular transport. Chapter 2 introduces the single molecule localization techniques that improve the spatial resolution of light microscopy. This chapter emphasizes the Stochastic Optical Reconstruction Microscopy (STORM) technique, which I used to study the organization of microtubule based motor proteins around lysosomes as well as the fusion and fission of mitochondria. Chapter 3 describes the development of two new techniques: (i) the use of microfluidic devices to improve the throughput of correlative live-cell and super-resolution microscopy, thus allowing to observe rare events and (ii) sequential multi-color imaging that increases the number of colors that can be imaged with STORM. Chapter 4 focuses on the biological application of sequential multicolor imaging to study the 3D organization of dynein and kinesin on lysosomal membranes. Conclusions and Future Perspectives are provided in Chapter 5. |
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