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
Diepold A
(2017)
A dynamic and adaptive network of cytosolic interactions governs protein export by the T3SS injectisome.
in Nature communications
Portwich FL
(2022)
A Highly Fluorescent Dinuclear Aluminium Complex with Near-Unity Quantum Yield.
in Angewandte Chemie (International ed. in English)
Frawley AT
(2023)
A Photoswitchable Solvatochromic Dye for Probing Membrane Ordering by RESOLFT Super-resolution Microscopy.
in Chemphyschem : a European journal of chemical physics and physical chemistry
Barbotin A
(2019)
Adaptive optics allows STED-FCS measurements in the cytoplasm of living cells.
in Optics express
Gutowska-Owsiak D
(2020)
Addressing Differentiation in Live Human Keratinocytes by Assessment of Membrane Packing Order.
in Frontiers in cell and developmental biology
Eggeling C
(2018)
Advances in bioimaging-challenges and potentials
in Journal of Physics D: Applied Physics
Rujas E
(2020)
Affinity for the Interface Underpins Potency of Antibodies Operating In Membrane Environments.
in Cell reports
Urbancic I
(2021)
Aggregation and mobility of membrane proteins interplay with local lipid order in the plasma membrane of T cells.
in FEBS letters
Kühne M
(2023)
Analysis of HDACi-Coupled Nanoparticles: Opportunities and Challenges.
in Methods in molecular biology (Clifton, N.J.)
Russell RA
(2017)
Astrocytes Resist HIV-1 Fusion but Engulf Infected Macrophage Material.
in Cell reports
Van Der Velde JHM
(2018)
Author Correction: A simple and versatile design concept for fluorophore derivatives with intramolecular photostabilization.
in Nature communications
Barbotin A
(2020)
Background Reduction in STED-FCS Using a Bivortex Phase Mask.
in ACS photonics
Barbotin A
(2020)
Background reduction in STED-FCS using coherent-hybrid STED
Kühne M
(2021)
Biocompatible sulfated valproic acid-coupled polysaccharide-based nanocarriers with HDAC inhibitory activity.
in Journal of controlled release : official journal of the Controlled Release Society
Santos AM
(2018)
Capturing resting T cells: the perils of PLL.
in Nature immunology
Felce JH
(2018)
CD45 exclusion- and cross-linking-based receptor signaling together broaden FceRI reactivity.
in Science signaling
Büttner M
(2021)
Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.
in Chembiochem : a European journal of chemical biology
Torralba J
(2020)
Cholesterol Constrains the Antigenic Configuration of the Membrane-Proximal Neutralizing HIV-1 Epitope.
in ACS infectious diseases
Tröger J
(2020)
Comparison of Multiscale Imaging Methods for Brain Research.
in Cells
Reina F
(2018)
Complementary studies of lipid membrane dynamics using iSCAT and super-resolved fluorescence correlation spectroscopy.
in Journal of physics D: Applied physics
Lagerholm BC
(2017)
Convergence of lateral dynamic measurements in the plasma membrane of live cells from single particle tracking and STED-FCS.
in Journal of physics D: Applied physics
Xiong Y
(2018)
Correction: Spironaphthoxazine switchable dyes for biological imaging.
in Chemical science
Van Der Velde JHM
(2017)
Corrigendum: A simple and versatile design concept for fluorophore derivatives with intramolecular photostabilization.
in Nature communications
Van Der Velde JHM
(2017)
Corrigendum: A simple and versatile design concept for fluorophore derivatives with intramolecular photostabilization.
in Nature communications
Sezgin E
(2020)
Creating Supported Plasma Membrane Bilayers Using Acoustic Pressure.
in Membranes
Fritzsche M
(2017)
Cytoskeletal actin dynamics shape a ramifying actin network underpinning immunological synapse formation.
in Science advances
Colin-York H
(2019)
Cytoskeletal actin patterns shape mast cell activation.
in Communications biology
Colin-York H
(2019)
Cytoskeletal Control of Antigen-Dependent T Cell Activation.
in Cell reports
Ebrahimi V
(2023)
Deep learning enables fast, gentle STED microscopy.
in bioRxiv : the preprint server for biology
Galiani S
(2022)
Diffusion and interaction dynamics of the cytosolic peroxisomal import receptor PEX5.
in Biophysical reports
Schneider F
(2017)
Diffusion of lipids and GPI-anchored proteins in actin-free plasma membrane vesicles measured by STED-FCS.
in Molecular biology of the cell
Clausen MP
(2017)
Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy.
in Journal of physics D: Applied physics
Colin-York H
(2017)
Dissection of mechanical force in living cells by super-resolved traction force microscopy.
in Nature protocols
Urbancic I
(2023)
Do lipids tune B cell signaling?
in Nature chemical biology
Eggeling C
(2018)
Editorial.
in Methods (San Diego, Calif.)
Portwich F
(2022)
Ein stark fluoreszierender zweikerniger Aluminiumkomplex mit nahezu 100 %iger Quantenausbeute**
in Angewandte Chemie
Pereno V
(2017)
Electroformation of Giant Unilamellar Vesicles on Stainless Steel Electrodes.
in ACS omega
Chojnacki J
(2017)
Envelope glycoprotein mobility on HIV-1 particles depends on the virus maturation state.
in Nature communications
Carravilla P
(2022)
Erratum: Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.
in Biophysical reports
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
Yang T
(2023)
Excited-State Dynamics in Borylated Arylisoquinoline Complexes in Solution and in cellulo.
in Chemistry (Weinheim an der Bergstrasse, Germany)
Schmidt F
(2020)
Flotillin-Dependent Membrane Microdomains Are Required for Functional Phagolysosomes against Fungal Infections.
in Cell reports
Reglinski K
(2020)
Fluidity and Lipid Composition of Membranes of Peroxisomes, Mitochondria and the ER From Oleic Acid-Induced Saccharomyces cerevisiae.
in Frontiers in cell and developmental biology
Carravilla P
(2020)
Fluorescence Microscopy of the HIV-1 Envelope.
in Viruses
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 |