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
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
Hufsky F
(2023)
The International Virus Bioinformatics Meeting 2023.
in Viruses
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
Jenkins E
(2018)
Reconstitution of immune cell interactions in free-standing membranes.
in Journal of cell science
Joseph J
(2024)
Nanoscale chemical characterization of secondary protein structure of F-Actin using mid-infrared photoinduced force microscopy (PiF-IR).
in Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy
Koerfer A
(2024)
Influence of different surface cleaning methods on STED-FCS and scanning STED-FCS calibration measurements.
in Journal of microscopy
Kokot B
(2021)
How to control fluorescent labeling of metal oxide nanoparticles for artefact-free live cell microscopy
in Nanotoxicology
Koller T
(2017)
Rituximab capping triggers intracellular reorganization of B cells
in Matters
Kowalczuk K
(2024)
Self-Degrading Multifunctional PEG-Based Hydrogels-Tailormade Substrates for Cell Culture.
in Macromolecular bioscience
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
Krstic A
(2025)
Enhancing entangled two-photon absorption of Nile Red via temperature-controlled SPDC
in APL Quantum
Kühne M
(2023)
Analysis of HDACi-Coupled Nanoparticles: Opportunities and Challenges.
in Methods in molecular biology (Clifton, N.J.)
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
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
Lyman E
(2018)
From Dynamics to Membrane Organization: Experimental Breakthroughs Occasion a "Modeling Manifesto".
in Biophysical journal
Lühr JJ
(2020)
Maturation of Monocyte-Derived DCs Leads to Increased Cellular Stiffness, Higher Membrane Fluidity, and Changed Lipid Composition.
in Frontiers in immunology
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
Pereno V
(2017)
Electroformation of Giant Unilamellar Vesicles on Stainless Steel Electrodes.
in ACS omega
Pinkwart K
(2019)
Nanoscale dynamics of cholesterol in the cell membrane
Pinkwart K
(2019)
Nanoscale dynamics of cholesterol in the cell membrane.
in The Journal of biological chemistry
Plochberger B
(2017)
HDL particles incorporate into lipid bilayers - a combined AFM and single molecule fluorescence microscopy study
in Scientific Reports
Polley A
(2017)
Glycosylation and Lipids Working in Concert Direct CD2 Ectodomain Orientation and Presentation
in The Journal of Physical Chemistry Letters
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)
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
Reina F
(2021)
TRAIT2D: a Software for Quantitative Analysis of Single Particle Diffusion Data.
in F1000Research
Reina F
(2018)
Complementary studies of lipid membrane dynamics using iSCAT and super-resolved fluorescence correlation spectroscopy.
in Journal of physics D: Applied physics
Reinhard J
(2023)
Laboratory-Based Correlative Soft X-ray and Fluorescence Microscopy in an Integrated Setup
in Microscopy and Microanalysis
Rissanen S
(2017)
Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin.
in Frontiers in physiology
Rujas E
(2020)
Affinity for the Interface Underpins Potency of Antibodies Operating In Membrane Environments
in Cell Reports
Russell RA
(2017)
Astrocytes Resist HIV-1 Fusion but Engulf Infected Macrophage Material.
in Cell reports
Santos AM
(2018)
Capturing resting T cells: the perils of PLL.
in Nature immunology
Schermelleh L
(2019)
Super-resolution microscopy demystified.
in Nature cell biology
Schmidt F
(2020)
Flotillin-Dependent Membrane Microdomains Are Required for Functional Phagolysosomes against Fungal Infections.
in Cell reports
Schneider F
(2020)
High photon count rates improve the quality of super-resolution fluorescence fluctuation spectroscopy.
in Journal of physics D: Applied physics
Schneider F
(2021)
Influence of nanobody binding on fluorescence emission, mobility, and organization of GFP-tagged proteins.
in iScience
Schneider F
(2018)
Nanoscale Spatiotemporal Diffusion Modes Measured by Simultaneous Confocal and Stimulated Emission Depletion Nanoscopy Imaging
in Nano Letters
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
Schneider F
(2018)
Statistical Analysis of Scanning Fluorescence Correlation Spectroscopy Data Differentiates Free from Hindered Diffusion.
in ACS nano
Schneider KRA
(2020)
Intracellular Photophysics of an Osmium Complex bearing an Oligothiophene Extended Ligand.
in Chemistry (Weinheim an der Bergstrasse, Germany)