Technique - (24) Live-cell fluorescence imaging

Type: Experimental

Description: Time-lapse epifluorescence or TIRF imaging of labeled proteins in living cells.

Department(s)/lab(s): School of Physics (joint with Biochemistry and Pharmacology) | Hinde Laboratory (Cell Nucleus Biophysics) @ UMelb
Summary:

Hinde is a fluorescence-fluctuation physicist embedded in cell biology: she uses pair-correlation function analysis, number-and-brightness, phasor-FLIM and FRET to read out chromatin compaction, protein-chromatin binding dynamics and nucleocytoplasmic transport in living nuclei, at spatial and temporal scales that conventional imaging averages away. The programme is a technique-pushing one — the emphasis is on extracting nanoscale structural information from photon statistics rather than on brute-force localisation — and it is now being coupled to quantum sensing through her QUBIC investigatorship, where the goal is to combine fluorescence readouts with NV-based magnetic and spin-noise contrast in the same cell. Positioned against the established body of NV-ensemble quantum sensing work — DEER, nanoscale NMR and T1 relaxometry protocols operating at pT/sqrt(Hz) field sensitivity — her role in QUBIC is to supply the cell-biological questions and the correlative optical readouts that make pT/sqrt(Hz)-class ensemble sensing biologically interpretable. Preferred attribute present: lifetime- and orientation-resolved methods pushing past the usual resolution limits.

Department(s)/lab(s): Physics & Astronomy – Biophysics & London Centre for Nanotechnology | Hoogenboom Lab (High-Speed AFM and Nanoscale Biophysics) @ UCL
Summary:

Hoogenboom leads a biophysics group at UCL specializing in high-speed atomic force microscopy. Research directions: (1) High-speed AFM — imaging conformational dynamics of DNA, proteins (including membrane channels), and chromatin at ms time resolution and sub-nm spatial resolution in aqueous conditions; (2) Nuclear pore complex — mapping transport selectivity and structure of NPCs in native nuclear envelopes using AFM; (3) Antimicrobial mechanisms — imaging membrane disruption by antimicrobial peptides in real time; (4) AFM-based force spectroscopy — measuring single-molecule interaction forces in chromatin and protein assemblies. Strong relevance to biological sensing at the single-molecule level.

Department(s)/lab(s): Physics & Astronomy – Biophysics | Jones Lab (Optical Tweezers Biophysics) @ UCL
Summary:

Jones's group develops optical tweezers instrumentation for biological applications. Research directions: (1) Single-cell mechanics — using optical traps to apply calibrated forces to cells and measure viscoelastic properties relevant to cancer invasion and immune response; (2) Motor protein biophysics — measuring force-velocity curves of kinesin/myosin motors at the single-molecule level; (3) Optical sorting — holographic optical tweezers for cell sorting by mechanical phenotype; (4) Instrument development — fast-switching AOD-based traps, quantitative phase imaging combined with force measurement. Sensitive to pN forces, combining biosensing with fundamental biophysics.

Department(s)/lab(s): Neurobiology | Kozorovitskiy Laboratory @ Northwestern
Summary:

Prof. Kozorovitskiy (Neurobiology) studies neuromodulation and plasticity in the striatum and basal ganglia, with a distinctive emphasis on developing and applying advanced optical imaging methods. Imaging technique innovations: (1) Oblique plane illumination (OPI / scanned oblique plane illumination, SOPi) microscopy — a single-objective light-sheet technique achieving tilt-invariant volumetric imaging for rapid 3D capture of fluorescently labeled neural structures without mechanical tilting; (2) Two-photon fluorescence imaging and two-photon glutamate/neuromodulator photorelease for single-synapse resolution in live tissue; (3) Near-infrared genetically-encoded calcium indicators (with Verkhusha group) for in vivo multi-color neural recording with reduced photobleaching. The lab's technical contributions are centered on extending the spatial and volumetric resolution of live-tissue fluorescence imaging. Irving M. Klotz Research Professor of Neurobiology; Beckman Young Investigator 2015.

Department(s)/lab(s): Yusuf Hamied Department of Chemistry | The Lee Lab – Biophysical Chemistry @ Cambridge
Summary:

Lee leads TheLeeLab at Cambridge Chemistry, focused on developing cutting-edge biophysical single-molecule fluorescence methods to answer fundamental biological questions. Two major thrusts: (1) 3D super-resolution microscopy instrument development — the lab pioneered single-molecule light field microscopy (SMLFM) using a microlens array in the back focal plane, achieving ~10× speed improvement over double-helix PSF for volumetric imaging; also develops vortex light field microscopy (VLFM) for simultaneous 25 nm spatial / 3 nm spectral precision; (2) Biological applications — studying T-cell receptor signalling at the nanoscale (distribution of TCRs, microvilli-mediated close contacts), histone assembly during DNA replication and repair in fission yeast, and PSD-95 nanoclusters in mouse brain using 3D SMLM. A job posting (PDRA) was active in 2025 for T-cell imaging work with super-resolution and Fourier light-field microscopy.

Department(s)/lab(s): Physics | Leifer Lab @ Princeton
Summary:

Leifer develops closed-loop optical instrumentation that simultaneously records brain-wide calcium activity and delivers single-neuron optogenetic perturbations in freely moving C. elegans, building functional atlases of signal propagation and studying how whole-brain neural dynamics generate behavior. His group's whole-brain, cellular-resolution imaging in unrestrained animals is a benchmark advanced-microscopy approach for linking neural dynamics to behavior.

Department(s)/lab(s): Biology / Institute of Molecular Biology (IMB) | Lemke Lab - Synthetic Biophysics @ JGU
Summary:

Lemke holds the chair of Synthetic Biophysics at JGU and is adjunct director at the Institute of Molecular Biology. The group's signature is combining genetic code expansion -- installing non-canonical amino acids so a dye can be clicked onto one chosen residue -- with single-molecule fluorescence: smFRET on intrinsically disordered proteins, super-resolution imaging of the nuclear pore complex and its FG-nucleoporin permeability barrier, and engineered membraneless organelles used as designer compartments in living cells. The result is single-molecule-resolution measurement of conformational dynamics and phase behaviour inside cells rather than in vitro. Relative to the established NV-ensemble quantum-sensing playbook (DEER, nanoscale NMR, T1 relaxometry at pT/sqrt(Hz) ensemble sensitivity), this is the strongest biosensing/advanced-microscopy host in Mainz: the labelling chemistry is precisely what a quantum-sensing postdoc would need to attach nanodiamonds or spin labels to a defined protein site, and the group already operates at the single-molecule sensitivity limit optically. Large, well-funded, internationally recruiting group.

Department(s)/lab(s): Department of Physics, 2nd Institute of Physics | Monzel Group - Biophysics and Biophotonics (2. Physikalisches Institut) @ Stuttgart
Summary:

Monzel holds the biophysics/biophotonics professorship at Stuttgart's 2nd Institute of Physics. The group develops multiparametric imaging spectroscopy and high-resolution light microscopy -- combining super-resolution, fluorescence-fluctuation and lifetime-resolved methods -- to read out several observables at once in living cells and in biomimetic model membranes, and pairs this with magnetic nanoparticles used to apply and sense forces on cell-surface receptors (magnetogenetic control of signalling). Single-molecule analysis inside cells is an explicit focus. Relative to the established NV-ensemble quantum-sensing playbook (DEER, nanoscale NMR, T1 relaxometry at pT/sqrt(Hz) ensemble sensitivity), this is the closest thing at Stuttgart to a natural biological host for in-cell quantum sensing: the group already does single-molecule-resolution live-cell imaging and already works with magnetic nanoparticles, so nanodiamond relaxometry/thermometry would slot in with the readout stack it already runs. Relatively new appointment -- good moment to join.

Department(s)/lab(s): Chemistry – Photon Science Institute | Natrajan Group (Lanthanide Photophysics and Biosensing) @ Manchester
Summary:

Natrajan's group develops luminescent lanthanide complexes for chemical and biological sensing. Research directions: (1) Time-gated lanthanide luminescence sensing — long-lifetime Eu3+, Tb3+, and Yb3+ complexes with millisecond emission lifetimes for background-free sensing in cells and tissue; (2) Intracellular sensing — luminescent probes for sensing O2, pH, viscosity, and specific enzymes inside living cells with spatiotemporal resolution; (3) Chiral discrimination — circularly polarized luminescence (CPL) from Eu3+ complexes for enantioselective sensing; (4) Responsive probes — switchable lanthanide complexes as ratiometric sensors for biomedical imaging. The long-lifetime emission enables time-gating strategies analogous to quantum sensing protocols.

Department(s)/lab(s): Physics & Astronomy – Biophysics | Nguyen Lab (Nanomaterials for Biosensing) @ UCL
Summary:

Nguyen's group at UCL (based at Royal Institution) focuses on magnetic and fluorescent nanoparticles for biomedical sensing and therapy. Research directions: (1) Magnetic nanoparticle synthesis — iron oxide (SPION) and other magnetic nanoparticles with controlled size, shape, and surface chemistry for MRI contrast and magnetic hyperthermia; (2) Biosensing platforms — functionalized nanoparticles as MRI-detectable sensors for specific biomolecular targets; magnetic particle imaging (MPI) for real-time tracking; (3) Plasmonic nanoparticles — gold nanoparticles for optical biosensing and photothermal therapy; (4) Fluorescent nanoparticles — QD- and dye-conjugated probes for live-cell imaging. Relevant to quantum sensing through magnetic nanoparticle platforms.