Technique - (55) Single-molecule fluorescence spectroscopy

Type: Experimental

Description: TIRF or confocal detection of fluorophore-labeled molecules; FRET, conformational dynamics.

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 | Liu Group - Smart Nanoplasmonics (2. Physikalisches Institut) @ Stuttgart
Summary:

Liu's group sits at the junction of DNA nanotechnology and nanophotonics: DNA-origami-templated plasmonic assemblies, reconfigurable artificial nanomachines whose motion is read out optically (chiral plasmonics, FRET), and, increasingly, synthetic-cell systems -- DNA-based pores and a programmable DNA-origami nanosyringe for directed membrane translocation, the latter published jointly with Nussberger's biophysics group at Stuttgart. The through-line is building nanoscale machines that both actuate and report. Relative to the established NV-ensemble quantum-sensing playbook (DEER, nanoscale NMR, T1 relaxometry at pT/sqrt(Hz) ensemble sensitivity), the relevance is on the biosensing axis: this is the group that can put a nanoscale probe exactly where you want it on or through a membrane, which is the delivery problem that in-cell quantum sensing keeps running into. Preferred-attribute note: nanofabrication is heavily used, but the emphasis is on single-molecule optical readout rather than device manufacture per se.

Department(s)/lab(s): Chemistry | PPSM - Single-Molecule Photochemistry (Metivier) @ ENSPS
Summary:

Metivier (PPSM) studies photochromic and fluorescent molecules at the single-molecule level - photoswitching kinetics, energy transfer and orientation-resolved imaging - underpinning super-resolution (RESOLFT/STORM-type) probes and molecular sensors. In the broader landscape of NV-centre ensemble quantum sensing (DEER, nano-NMR, T1 relaxometry) operating near pT/sqrt(Hz) sensitivity, this work is paralleled by molecular photoswitches enabling optical super-resolution.

Department(s)/lab(s): Chemistry | Moerner Lab @ Stanford
Summary:

Nobel laureate W. E. Moerner, who first detected and studied single molecules optically, now develops engineered point-spread-function and orientation-resolved single-molecule localization microscopy methods to track individual biomolecules and their rotational dynamics in cells with nanometer precision, well beyond the optical diffraction limit.

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): Physics – Institute for Quantum Electronics | Nanoscale Quantum Optics Group (Murthy) @ ETH Zurich
Summary:

Murthy leads the Nanoscale Quantum Optics group at ETH, studying light-matter interactions in nanostructures to engineer novel quantum states of light. Research directions: (1) Photon-photon interactions — achieving strong effective photon-photon interactions via coupling to quantum emitters in 2D materials and optical nanocavities; exploring photonic Mott insulators and collective quantum phases of light; (2) 2D semiconductor quantum emitters — localized excitons in TMD heterostructures as sources of single photons and entangled photon pairs; (3) Quantum light from cavities — engineering photon statistics and squeezing using cavity-QED with 2D materials; (4) Ultrafast quantum optics — attosecond-scale probing of light-matter entanglement. New group as of ~2023.

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): 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): Institute of Biomaterials and Biomolecular Systems | Nussberger Lab - Biophysics @ Stuttgart
Summary:

Nussberger holds the biophysics chair at Stuttgart's Institute of Biomaterials and Biomolecular Systems. The group studies how proteins cross and insert into membranes -- mitochondrial protein translocases (TOM complex), apoptosis-related pore formation -- using single-channel electrophysiology, single-molecule fluorescence and structural methods, and has pushed this into an explicit nanopore/biosensing line: engineered protein and DNA-based pores as single-molecule sensors, including the DNA-origami nanosyringe for directed membrane translocation published with Na Liu's group. Relative to the established NV-ensemble quantum-sensing playbook (DEER, nanoscale NMR, T1 relaxometry at pT/sqrt(Hz) ensemble sensitivity), the relevance is the readout channel: nanopore sensing is the electrical single-molecule counterpart to optical single-molecule detection, and the group's membrane expertise is exactly what an in-cell quantum-sensing project needs when the question becomes how to get the probe across a bilayer.

Department(s)/lab(s): Biochemistry and Molecular Biology | Rock Lab @ UChicago
Summary:

Rock builds custom single-molecule fluorescence microscopes and optical tweezers to directly watch individual myosin motors move along the actin cytoskeleton in vitro and in living cells, quantifying motor stoichiometry, force generation, and navigation rules that organize cell shape and motility. Where NV-ensemble quantum sensors read out spin ensembles magnetically at pT/sqrt(Hz) sensitivity via DEER/NMR/T1 protocols, Rock's approach achieves single-fluorophore and single-motor mechanical/positional resolution using all-optical single-molecule methods.