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
Amaro M
(2017)
Laurdan and Di-4-ANEPPDHQ probe different properties of the membrane.
in Journal of physics D: Applied physics
Ando T
(2018)
The 2018 correlative microscopy techniques roadmap.
in Journal of physics D: Applied physics
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
Barbotin A
(2020)
z-STED Imaging and Spectroscopy to Investigate Nanoscale Membrane Structure and Dynamics.
in Biophysical journal
Barbotin A
(2019)
Adaptive optics allows STED-FCS measurements in the cytoplasm of living cells.
in Optics express
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
Basu S
(2018)
FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells.
in Nature communications
Bedard M
(2019)
Sterile activation of invariant natural killer T cells by ER-stressed antigen-presenting cells.
in Proceedings of the National Academy of Sciences of the United States of America
Behnke M
(2024)
PEtOxylated polyesteramide nanoparticles for the delivery of anti-inflammatory drugs
in Materials Today Chemistry
BĂĽttner M
(2021)
Challenges of Using Expansion Microscopy for Super-resolved Imaging of Cellular Organelles.
in Chembiochem : a European journal of chemical biology
Carravilla P
(2022)
Erratum: Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.
in Biophysical reports
Carravilla P
(2020)
Fluorescence Microscopy of the HIV-1 Envelope.
in Viruses
Carravilla P
(2019)
Molecular recognition of the native HIV-1 MPER revealed by STED microscopy of single virions.
in Nature communications
Carravilla P
(2021)
Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.
in Biophysical reports
Carugo D
(2017)
Modulation of the molecular arrangement in artificial and biological membranes by phospholipid-shelled microbubbles.
in Biomaterials
Chojnacki J
(2017)
Envelope glycoprotein mobility on HIV-1 particles depends on the virus maturation state.
in Nature communications
Chojnacki J
(2018)
Zooming in On Virus Surface Protein Mobility
in Future Virology
Chojnacki J
(2018)
Super-resolution fluorescence microscopy studies of human immunodeficiency virus.
in Retrovirology
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
Colin-York H
(2019)
Cytoskeletal Control of Antigen-Dependent T Cell Activation
in Cell Reports
Colin-York H
(2019)
Cytoskeletal actin patterns shape mast cell activation.
in Communications biology
Dasgupta A
(2024)
Effects and avoidance of photoconversion-induced artifacts in confocal and STED microscopy.
in Nature methods
De Angelis G
(2024)
Homogeneous large field-of-view and compact iSCAT-TIRF setup for dynamic single molecule measurements
in Optics Express
Diepold A
(2017)
A dynamic and adaptive network of cytosolic interactions governs protein export by the T3SS injectisome.
in Nature communications
Ebrahimi V
(2023)
Author Correction: Deep learning enables fast, gentle STED microscopy.
in Communications biology
Ebrahimi V
(2023)
Deep learning enables fast, gentle STED microscopy.
in Communications biology
Ebrahimi V
(2023)
Deep learning enables fast, gentle STED microscopy
Eggeling C
(2017)
Macrophages: micromanagers of antagonistic signaling nanoclusters.
in The Journal of cell biology
Eggeling C
(2018)
Editorial
in Methods
Eggeling C
(2018)
Advances in bioimaging-challenges and potentials
in Journal of Physics D: Applied Physics
Favard C
(2019)
HIV-1 Gag specifically restricts PI(4,5)P2 and cholesterol mobility in living cells creating a nanodomain platform for virus assembly.
in Science advances
Felce JH
(2018)
CD45 exclusion- and cross-linking-based receptor signaling together broaden FceRI reactivity.
in Science signaling
Franke C
(2024)
Electrochemistry meets Photophysics for single-molecule localization
in Nature Photonics
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
Frawley AT
(2020)
Super-resolution RESOLFT microscopy of lipid bilayers using a fluorophore-switch dyad.
in Chemical science
Fritzsche M
(2017)
Cytoskeletal actin dynamics shape a ramifying actin network underpinning immunological synapse formation.
in Science advances
Fritzsche M
(2017)
Self-organizing actin patterns shape membrane architecture but not cell mechanics.
in Nature communications
Galiani S
(2022)
Diffusion and interaction dynamics of the cytosolic peroxisomal import receptor PEX5.
in Biophysical reports
Galiani S
(2023)
Super-resolution microscopy and studies of peroxisomes
in Biological Chemistry
Gutowska-Owsiak D
(2020)
Addressing Differentiation in Live Human Keratinocytes by Assessment of Membrane Packing Order.
in Frontiers in cell and developmental biology
Gutowska-Owsiak D
(2018)
Orchestrated control of filaggrin-actin scaffolds underpins cornification.
in Cell death & disease