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
Büttner M
(2021)
Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.
in Chembiochem : a European journal of chemical biology
Frawley AT
(2020)
Super-resolution RESOLFT microscopy of lipid bilayers using a fluorophore-switch dyad.
in Chemical science
Xiong Y
(2018)
Spironaphthoxazine switchable dyes for biological imaging.
in Chemical science
Xiong Y
(2018)
Correction: Spironaphthoxazine switchable dyes for biological imaging.
in Chemical science
Schneider KRA
(2020)
Intracellular Photophysics of an Osmium Complex bearing an Oligothiophene Extended Ligand.
in Chemistry (Weinheim an der Bergstrasse, Germany)
Yang T
(2023)
Excited-State Dynamics in Borylated Arylisoquinoline Complexes in Solution and in cellulo.
in Chemistry (Weinheim an der Bergstrasse, Germany)
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
Colin-York H
(2019)
Cytoskeletal actin patterns shape mast cell activation.
in Communications biology
Steinkühler J
(2019)
Mechanical properties of plasma membrane vesicles correlate with lipid order, viscosity and cell density.
in Communications biology
Hertzog J
(2018)
Infection with a Brazilian isolate of Zika virus generates RIG-I stimulatory RNA and the viral NS5 protein blocks type I IFN induction and signaling.
in European journal of immunology
Reina F
(2021)
TRAIT2D: a Software for Quantitative Analysis of Single Particle Diffusion Data.
in F1000Research
Urbancic I
(2021)
Aggregation and mobility of membrane proteins interplay with local lipid order in the plasma membrane of T cells.
in FEBS letters
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
Azbazdar Y
(2019)
More Favorable Palmitic Acid Over Palmitoleic Acid Modification of Wnt3 Ensures Its Localization and Activity in Plasma Membrane Domains.
in Frontiers in cell and developmental biology
Gutowska-Owsiak D
(2020)
Addressing Differentiation in Live Human Keratinocytes by Assessment of Membrane Packing Order.
in Frontiers in cell and developmental biology
Lühr J
(2020)
Maturation of Monocyte-Derived DCs Leads to Increased Cellular Stiffness, Higher Membrane Fluidity, and Changed Lipid Composition
in Frontiers in Immunology
Rissanen S
(2017)
Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin.
in Frontiers in physiology
Chojnacki J
(2018)
Zooming in on virus surface protein mobility
in Future Virology
Insausti S
(2022)
Functional Delineation of a Protein-Membrane Interaction Hotspot Site on the HIV-1 Neutralizing Antibody 10E8.
in International journal of molecular sciences
Schneider F
(2021)
Influence of nanobody binding on fluorescence emission, mobility, and organization of GFP-tagged proteins.
in iScience
Jenkins E
(2018)
Reconstitution of immune cell interactions in free-standing membranes.
in Journal of cell science
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
Koerfer A
(2023)
Influence of different surface cleaning methods on STED-FCS and scanning STED-FCS calibration measurements
in Journal of Microscopy
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 |