Fluorescence Spectroscopy & Single Molecule Fluorescence
Application
The bespoke equipment allows collection of single-molecule fluorescence from soluble as well as immobilized samples in vitro with temporal resolution from nanoseconds to minutes. The confocal instrument is equipped for 480 and 561 nm pulsed interleaved excitation suitable for FRET (Forster resonance energy transfer) intensity and fluorescence lifetime measurements. While the TIRFM instrument is equipped with four excitation wavelengths (405, 488, 561 and 640 nm). The multi-parameter fluorescence detection enables us to obtain correlation functions and other dynamic information on complex, fluorescently labelled systems. Applications range from structural and dynamic characterization of ensemble conformers of proteins, nucleic acids and their assemblies to following membrane protein dynamics and virus assembly in real time. Open design allows continuous modification and customization to accommodate e.g. concurrent electrochemical measurements or additional fluorescence or scattering channels.
Equipment
Horiba Quantamaster 8075 spectrofluorometer (located in L. C. Miall 10.22) equipped with Whitelase 420 pulsed white laser (with frequency doubler for UV excitation of e.g. Tryptophan fluorescence, variable repletion rate 0.1 to 10 MHz) and time-correlated detection in the visible and near infrared region. This instrument has titrator accessory, thermoelectric temperature control, integrating sphere, solid sample holder and motorized polarizers for anisotropy measurements. The setup is suitable for quantitative ensemble measurements of quantum yield and lifetime (e.g. quantum dot and fluorescent dye characterization), affinity titrations (e.g using anisotropy) and characterization of up-converting quantum dots. Routine measurements of emission and excitation are readily available across the extended spectra range 200-1600 nm.
Location
Garstang 10.135 (Single Molecule Laboratory)
Contact
Dr Roman Tuma
School of Molecular & Cellular Biology
Email: r.tuma@leeds.ac.uk
OR
Dr Matt Watson
School of Molecular & Cellular Biology
Email: m.a.watson@leeds.ac.uk