Technique - (21) Live-cell fluorescence imaging

Type: Experimental

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

Department(s)/lab(s): EMBL Australia Node in Single Molecule Science, UNSW Medicine and Health | Ananthanarayanan Cell Biology and Advanced Microscopy Group @ UNSW
Summary:

Ananthanarayanan was awarded the Royal Microscopical Society Life Sciences Award in 2025 for the use of novel microscopies in cell biology. Her group images individual motor proteins — dynein, kinesin — and the mitochondria they transport, in living cells, at single-molecule sensitivity, combining light-sheet and TIRF-class imaging with particle tracking to ask how organelle positioning and mitochondrial dynamics are controlled. The methodological emphasis is on getting single-molecule sensitivity inside a live cell rather than in vitro, which is the hard version of the problem. 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 — this is the closest thing at UNSW to a biological end-user for an in-cell quantum sensor: the mitochondrial systems she studies are precisely where NV nanodiamond thermometry and free-radical relaxometry at pT/sqrt(Hz) have been aimed, and she has the live-cell imaging infrastructure to validate any such measurement independently.

Department(s)/lab(s): EMBL Australia Node in Single Molecule Science, UNSW Medicine and Health | Molecular Machines Group (Boecking) @ UNSW
Summary:

Boecking leads the Molecular Machines Group and is acting director of the EMBL Australia Node in Single Molecule Science. The group reconstitutes molecular machines — clathrin coat disassembly, HIV capsid assembly and uncoating, pore-forming toxins — and watches them work one molecule at a time by TIRF, interferometric scattering (mass photometry) and fluorescence fluctuation methods, resolving short-lived intermediates that ensemble kinetics averages into invisibility. He trained originally in surface chemistry and biosensors with Gooding, which gives the group unusual competence in engineering the surfaces these assays run on. 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 — the argument for single-molecule methods over ensemble ones is identical to the argument for pushing NV sensing below its pT/sqrt(Hz) ensemble regime: the interesting biology lives in heterogeneity and in transient states that averaging destroys. Strong methodological neighbour for a quantum-biosensing candidate.

Department(s)/lab(s): Imaging Physics (ImPhys) | Brinks Lab @ TU Delft
Summary:

Daan Brinks develops all-optical electrophysiology tools for neuroscience. His lab engineers genetically-encoded voltage indicators (GEVIs) and combines them with optogenetics to read out and control neural circuit activity. Key directions: (1) engineering bright, fast GEVIs with improved photostability and voltage sensitivity; (2) multiplexed all-optical neural circuit mapping; (3) identifying rare aggressive cancer cells using voltage-sensitive dyes. His voltage imaging approach represents cutting-edge biosensing at the intersection of photonics and neuroscience.

Department(s)/lab(s): Physics & Astronomy – Photon Science Institute | Bioimaging and Microscopy Group (Dickinson Group) @ Manchester
Summary:

Dickinson's group develops advanced optical microscopy methods for biological and biomedical imaging. Research directions: (1) STORM super-resolution microscopy — stochastic optical reconstruction for nanoscale imaging of biological structures at ~20 nm lateral resolution; imaging cytoskeletal dynamics, cellular organelles, and pathological structures; (2) Optical coherence tomography (OCT) — depth-resolved, label-free imaging for biomedical diagnostics (retinal, cardiovascular tissues); (3) Laser speckle imaging — blood flow and perfusion measurements in tissues; (4) Multiphoton microscopy — second harmonic generation (SHG) and two-photon for collagen structure imaging in connective tissues and cancer. Part of the Manchester Photon Science Institute biophotonics theme.

Department(s)/lab(s): EMBL Australia Node in Single Molecule Science, UNSW Medicine and Health | Gambin Single Molecule Biophysics Group @ UNSW
Summary:

Gambin was the first EMBL Australia group leader appointed to Single Molecule Science. His signature method combines cell-free protein expression with two-colour single-molecule coincidence and fluctuation spectroscopy, which sidesteps purification entirely: proteins are expressed, labelled and measured in lysate, an order of magnitude faster than conventional interaction assays. The biology is protein self-association and aggregation — alpha-synuclein in Parkinson's, cardiac and muscular disease proteins — where the size distribution of oligomers, not the mean, is the quantity of interest. 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 — the conceptual overlap with quantum biosensing is the insistence on distributions over averages, and his aggregation systems (paramagnetic-species-generating, redox-active amyloid) are a plausible target for T1-relaxometry-based NV detection at pT/sqrt(Hz) in the near term.

Department(s)/lab(s): Imaging Physics (ImPhys) | Geertsema Lab @ TU Delft
Summary:

Hylkje Geertsema uses single-molecule super-resolution fluorescence microscopy (TIRF, SMLM, PALM/STORM) to study DNA replication dynamics. Her lab visualises and quantifies individual replication proteins at replication forks in living cells to understand the kinetics and fidelity of DNA copying. Research focuses on measuring spatiotemporal dynamics of protein assemblies during DNA metabolism with nanometre resolution.

Department(s)/lab(s): Physics | Golding Lab @ UIUC
Summary:

Uses single-molecule fluorescence microscopy in live bacteria to study stochastic gene expression, chromosome organization, and cell-to-cell variability.

Department(s)/lab(s): Graduate School of Biomedical Engineering | Goldys Nanoscale Biophotonics Group @ UNSW
Summary:

Goldys was Deputy Director of the ARC Centre of Excellence for Nanoscale BioPhotonics and now leads a nanoscale biophotonics group in Biomedical Engineering. The programme is about extracting diagnostic information from very weak optical signals inside cells and tissue: luminescent and upconverting nanoparticle probes with long lifetimes that allow time-gated, background-free detection; hyperspectral unmixing of native cellular autofluorescence (NADH, FAD, porphyrins) as a completely label-free readout of cell state, which she has pushed toward clinical use in reproductive medicine and cancer; and nanoparticle-mediated therapy. 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 — time-gated luminescence and NV relaxometry are two solutions to the same problem — how to read a faint, specific signal out of an autofluorescent, optically hostile biological background — and her clinical translation experience is exactly the missing capability in most quantum-biosensing groups. Preferred attribute present: advanced/label-based imaging with a genuine human-application pathway.

Department(s)/lab(s): Physics | Laboratory for the Physics of Life @ Princeton
Summary:

Gregor's Laboratory for the Physics of Life builds custom quantitative microscopes (single-objective oblique-plane light-sheet, multicolor live-imaging, single-molecule transcription imaging) to make precision, physics-style measurements of gene expression, morphogen gradients, and chromatin dynamics in living Drosophila embryos and mammalian gastruloids. He is actively recruiting PhD students and postdocs with expertise in super-resolution imaging, nonlinear/ultrafast optics, and instrumentation development.

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.