Nano-Immunology
Lead Research Organisation:
University of Oxford
Department Name: UNLISTED
Abstract
Research by my group aims to unravel nanoscopic changes at the molecular level in living cells to characterise important molecular processes on the cell membrane as well as inside the cell during immunological reactions. Because many cellular responses lead to changes so subtle at the molecular level, studying them requires us to not only observe them with a superior spatial resolution but also to reach a sensitivity that is able to monitor single molecules over time and space. We are using the newest and most powerful super-resolution far-field microscopes (such as STED, RESOLFT or PALM/STORM) to image and analyse cellular structures and protein-protein and protein-lipid interactions at a level of fine detail that until now has not been possible due to the limited spatial resolution of conventional optical far-field microscopes. By combining these super-resolution microscopy techniques with single-molecule sensitive detection methods (such as fluorescence correlation spectroscopy) and fast spatio-temporal tracking tools we are able to see complex dynamic processes otherwise invisible because of the lower power of conventional far-field microscopy. These molecular interactions play an important role in the immune response to infection and cancer and so we intend to use and further develop these advanced microscopy techniques and apply them to gather new insights in immunological research.
Technical Summary
Single-molecule super-resolution microscopy of membrane dynamics: Many cellular responses lead to subtle changes on the molecular level, demanding not only for a superior spatial resolution of the analyzing method but also for the sensitivity to monitor single molecules over time and space. The combination of super-resolution optical fluorescence STED microscopy with single-molecule sensitive fluorescence-detection tools such as Fluorescence Correlation Spectroscopy (FCS) as well as the fast spatio-temporal tracking of single labeled molecules (single-particle tracking, SPT) allows for the disclosure of complex dynamic processes otherwise impeded by the limited spatial resolution of conventional far-field microscopy. For example, STED-FCS or SPT offer us the possibility to gain novel insights into important cellular processes, such as lipid-lipid, lipid-protein, and protein-protein interactions and the formation of so-called “lipid-rafts” in the cellular plasma membrane. These molecular interactions play an important role in the cellular immune response. We will therefore apply and further develop the STED-FCS and SPT microscopy techniques to highlight important molecular processes on the plasma membrane as well as inside the cell during immunological reactions. For example, these techniques will be used to shed new light on different molecular pathways triggered at the cell surface and intracellularly during antigen presentation by dendritic cells and T cell activation.
Organisations
People |
ORCID iD |
Christian Eggeling (Principal Investigator) |
Publications
Torralba J
(2020)
Cholesterol Constrains the Antigenic Configuration of the Membrane-Proximal Neutralizing HIV-1 Epitope.
in ACS infectious diseases
Schneider F
(2018)
Statistical Analysis of Scanning Fluorescence Correlation Spectroscopy Data Differentiates Free from Hindered Diffusion.
in ACS nano
Pereno V
(2017)
Electroformation of Giant Unilamellar Vesicles on Stainless Steel Electrodes.
in ACS omega
Barbotin A
(2020)
Background Reduction in STED-FCS Using a Bivortex Phase Mask.
in ACS photonics
Portwich F
(2022)
Ein stark fluoreszierender zweikerniger Aluminiumkomplex mit nahezu 100 %iger Quantenausbeute**
in Angewandte Chemie
Portwich FL
(2022)
A Highly Fluorescent Dinuclear Aluminium Complex with Near-Unity Quantum Yield.
in Angewandte Chemie (International ed. in English)
Mobarak E
(2018)
How to minimize dye-induced perturbations while studying biomembrane structure and dynamics: PEG linkers as a rational alternative.
in Biochimica et biophysica acta. Biomembranes
Galiani S
(2023)
Super-resolution microscopy and studies of peroxisomes.
in Biological chemistry
Carugo D
(2017)
Modulation of the molecular arrangement in artificial and biological membranes by phospholipid-shelled microbubbles.
in Biomaterials
Lyman E
(2018)
From Dynamics to Membrane Organization: Experimental Breakthroughs Occasion a "Modeling Manifesto"
in Biophysical Journal
Barbotin A
(2020)
z-STED Imaging and Spectroscopy to Investigate Nanoscale Membrane Structure and Dynamics.
in Biophysical journal
Sezgin E
(2017)
Polarity-Sensitive Probes for Superresolution Stimulated Emission Depletion Microscopy.
in Biophysical journal
Zhurgenbayeva G
(2023)
Quantification of biophysical properties of candidalysin, a fungal peptide toxin secreted by C. albicans during invasion
in Biophysical Journal
Carravilla P
(2022)
Erratum: Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.
in Biophysical reports
Galiani S
(2022)
Diffusion and interaction dynamics of the cytosolic peroxisomal import receptor PEX5.
in Biophysical reports
Carravilla P
(2021)
Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.
in Biophysical reports
Ebrahimi V
(2023)
Deep learning enables fast, gentle STED microscopy.
in bioRxiv : the preprint server for biology
Gutowska-Owsiak D
(2018)
Orchestrated control of filaggrin-actin scaffolds underpins cornification.
in Cell death & disease
Russell RA
(2017)
Astrocytes Resist HIV-1 Fusion but Engulf Infected Macrophage Material.
in Cell reports
Rujas E
(2020)
Affinity for the Interface Underpins Potency of Antibodies Operating In Membrane Environments.
in Cell reports
Schmidt F
(2020)
Flotillin-Dependent Membrane Microdomains Are Required for Functional Phagolysosomes against Fungal Infections.
in Cell reports
Colin-York H
(2019)
Cytoskeletal Control of Antigen-Dependent T Cell Activation.
in Cell reports
Tröger J
(2020)
Comparison of Multiscale Imaging Methods for Brain Research.
in Cells
Kretzer C
(2021)
Ethoxy acetalated dextran-based nanocarriers accomplish efficient inhibition of leukotriene formation by a novel FLAP antagonist in human leukocytes and blood.
in Cellular and molecular life sciences : CMLS
Related Projects
Project Reference | Relationship | Related To | Start | End | Award Value |
---|---|---|---|---|---|
MC_UU_00008/1 | 01/04/2017 | 31/03/2023 | £2,738,000 | ||
MC_UU_00008/2 | Transfer | MC_UU_00008/1 | 01/04/2017 | 31/03/2023 | £1,821,000 |
MC_UU_00008/3 | Transfer | MC_UU_00008/2 | 01/04/2017 | 31/03/2023 | £2,257,000 |
MC_UU_00008/4 | Transfer | MC_UU_00008/3 | 01/04/2017 | 31/03/2023 | £1,459,000 |
MC_UU_00008/5 | Transfer | MC_UU_00008/4 | 01/04/2017 | 31/03/2023 | £1,346,000 |
MC_UU_00008/6 | Transfer | MC_UU_00008/5 | 01/04/2017 | 31/03/2023 | £1,660,000 |
MC_UU_00008/7 | Transfer | MC_UU_00008/6 | 01/04/2017 | 31/03/2023 | £401,000 |
MC_UU_00008/8 | Transfer | MC_UU_00008/7 | 01/04/2017 | 31/03/2024 | £2,876,000 |
MC_UU_00008/9 | Transfer | MC_UU_00008/8 | 01/04/2017 | 31/03/2023 | £2,568,000 |
MC_UU_00008/10 | Transfer | MC_UU_00008/9 | 01/04/2017 | 31/03/2023 | £2,060,000 |
MC_UU_00008/11 | Transfer | MC_UU_00008/10 | 01/04/2017 | 31/03/2023 | £1,477,000 |