MULTIQUANTUM · Combining multi-photon microscopy and quantum dot technology to study molecular dynamics on single cells in vivo
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2006-03-01 → 2010-02-28
- EU contribution
- €1,403,791
- Participants
- 1
- Scheme
- EXT
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Results in brief
Final Activity Report Summary - MULTIQUANTUM (Combining multi-photon microscopy and quantum dot technology to study molecular dynamics on single cells in vivo)
Continuous lymphocyte trafficking through secondary lymphoid organs, such as spleen and peripheral lymph nodes (PLN), forms the basis for efficient immune surveillance by the adaptive immune system. Inside PLN, lymphocytes disseminate into T cell zones and B cell follicles where they screen dendritic cells (DCs) before departure via efferent lymphatic vessels. Together with adhesion receptors, lymphocyte recruitment from blood, interstitial migration and egress is regulated by G-protein coupled receptors, such as chemokine receptors and sphingosine-1-phosphate receptor 1. A major focus is the study of molecular mechanisms of lymphocyte migration to and within PLN, with a special emphasis on chemokine receptor signalling. To this end, we have applied novel imaging techniques for the study of these events, mainly two photon microscopy (2PM) in combination with quantum dot (QD) technology. Our results uncover the possibilities and pitfalls of the use of this novel technology, and highlight the usefulness of QDs for specific applications.
Data: CORDIS, © European Union
Project objective
Multi-photon intravital microscopy (MP-IVM) is a powerful imaging technique which allows following cellular migration and interactions on a single cell level deep inside solid tissue of alive, anesthetized mice. A prominent example is the recent descripti on of lymphocyte migration inside peripheral lymph nodes (PLN) during immune surveillance and responses to Antigen. Nevertheless, resolution of subcellular structures and molecular complexes is hampered by reduced brightness and rapid photobleaching of co mmonly used molecular fluorophores. Quantum dots (QD) are small fluorescent nanocrystals with superior optical properties compared to molecular fluorophores, such as resistance to photobleaching, large absorption spectra and narrow emission spectra, which can be controlled by crystal size. QD are intrinsically hydrophobic and need to be coated with specific peptides or other watersoluble substances for bioimaging. Here, we propose to use MP-IVM to observe interstitial migration and cellular interactions of lymphocytes labeled with commercially available QD for detection of surface molecule distribution. We furthermore propose to develop own application-specific QD coated with appropriate surface markers for internalization and labeling of intracellular s ignaling complexes. We expect this proposal to accomplish two major aims. First, we seek to explore the suitability of QD technology for MP-IVM. Second, we anticipate novel insights in subcellular distribution of specific molecular dynamics during inter stitial migration and cellular interactions.
Original text from CORDIS.
Participants
- UNIVERSITÄT BERN · BERNCoordinatorCity levelSwitzerland
Links
Data: CORDIS, © European Union