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Toward a single molecule study of asymmetric experiment

Abstract : The goal of this project is to develop novel tools to explore dynamic organizational processes in living cells, with unprecedented detail and sensitivity. In particular, we are interested in the process of asymmetric cell division (ACD), during which a mother cell divides asymmetrically into two different daughter cells. This process is a key mechanism in the generation of cellular diversity in organisms of all levels. During the course of ACD, prior to the separation of the mother cell into daughter cells, it needs to generate an asymmetric spatial distribution for some of its content. This symmetry-breaking process is called polarization i.e. the generation of a polarized structure. One of the ways to study polarization is to characterize the intra-cellular transport mechanisms that generate it. As a means to observe intra-cellular transport in detail, we employ single-molecule methods, which allow us to separately follow (or track) the movement of single molecules. By observing one event at a time we can obtain much more information about the process, compared to simultaneous measurements of the entire population. We are using novel and powerful fluorophores, quantum dots (QDs), which allow for single molecule detection and tracking. On the technical level, we developed methods to cope with certain technical difficulties associated with intra-cellular, single-molecule, tracking experiments. These include the delivery of such fluorophores into living cells, the targeting of specific molecules of interest, as well as control over the stoichiometry of QD-protein complexes. The project combines different disciplines, such as single molecule imaging, molecular biology, and primary cell culture, thereby covering almost all aspects of an in-vivo single-molecule experiment.
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Submitted on : Thursday, April 18, 2013 - 3:31:58 PM
Last modification on : Thursday, March 17, 2022 - 10:08:13 AM
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  • HAL Id : tel-00815355, version 1


Assa Sittner. Toward a single molecule study of asymmetric experiment. Biological Physics []. Université Pierre et Marie Curie - Paris VI, 2010. English. ⟨NNT : 2010PA066095⟩. ⟨tel-00815355⟩



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