Technique - (5) Polarization- and orientation-resolved fluorescence microscopy

Type: Experimental

Description: Fluorescence microscopy that resolves the orientation of individual fluorophores or fluorophore ensembles via polarization-modulated excitation and detection.

Department(s)/lab(s): Engineering | Institut Fresnel - Vector & Polarization Imaging Team @ CNRS
Summary:

Brasselet is a CNRS researcher at Institut Fresnel developing polarization- and orientation-resolved fluorescence microscopy, using controlled excitation and detection polarization states to map the 3D orientation and organization of fluorescent probes and biomolecular assemblies (e.g. lipid order, amyloid and cytoskeletal structures) at and beyond the single-molecule level, including recent work on the mathematical foundations of polarimetric microscopy.

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): Applied Physics, Molecular and Cellular Biology | Needleman Lab @ Harvard
Summary:

Needleman combines polarized-light microscopy, second-harmonic generation, single-molecule tracking, and fluorescence-lifetime (FLIM) metabolic imaging to study self-organization of the mitotic spindle and, in a clinically translated direction, non-invasive metabolic imaging of human oocytes and embryos for IVF viability assessment — an orientation- and lifetime-resolved imaging program with an active human-trial/clinical translation component.

Department(s)/lab(s): School of Physics | Roberts Optics and Meta-Optics Group @ UMelb
Summary:

Roberts leads Melbourne's optics group and is a chief investigator in the ARC Centre of Excellence for Transformative Meta-Optical Systems (TMOS). The work is about extracting information that conventional intensity imaging discards: metasurface-encoded point spread functions that recover the full polarisation state or quantitative phase in a single shot, subwavelength structures for edge enhancement and optical computing, and vectorial beam shaping. For a quantum-sensing candidate the relevant hook is that meta-optics is becoming the standard way to miniaturise the optical front end of NV, atomic-vapour and single-molecule sensors, and to add orientational sensitivity to imaging. 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 metasurface collection optics and polarisation-resolved detection schemes are being applied to improve photon collection efficiency and orientational discrimination in exactly the NV-ensemble geometries used for pT/sqrt(Hz) magnetometry. Preferred attribute present: orientation-resolved methods that push past standard resolution limits.

Department(s)/lab(s): School of Chemistry | Smith Time-Resolved Spectroscopy and Microspectroscopy Group @ UMelb
Summary:

Smith runs Melbourne's time-resolved fluorescence facility and specialises in the information channels most people throw away: fluorescence lifetime, anisotropy decay and its orientational content, and single-molecule photophysics, applied to organic semiconductors, energy-transfer systems and biological samples. The group builds its own confocal microspectroscopy instrumentation for time-resolved anisotropy imaging and single-molecule detection. 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 — lifetime- and orientation-resolved fluorescence is the principal orthogonal contrast mechanism to spin-based sensing, and his instrumentation is the natural correlative partner for NV-ensemble DEER/relaxometry experiments at pT/sqrt(Hz) that need an independent optical readout of the same specimen. Preferred attribute present: orientation- and lifetime-resolved methods.