Equipment
Fluorescence is a powerful analytical tool that combines the magnifying properties of light microscopy with the visualisation of fluorescence to monitor live real-time protein interactions, at steady state or after cellular ligand challenge. Currently, we have two state of the art laser point scanning confocal instruments (invert configuration) which support different areas of fluorescence microscopy.
Zeiss 880 LSCM
The system is equipped with a high-resolution galvo scanner and 3 channel QUASOR detector unit which is wavelenghth tuneable for custom spectral collection of emission light. The QUASOR consists of two side spectral green and red PMTs and a central GaAsP detector. A transmitted light PMT is available for bright field imaging. Images can be recorded up to a maximum 16 bit depth dynamic range. The red sensitive side PMT and GaAsP detector can also record fluorescence as a read out of photons which is more advantageous than analog integration read out because much higher signal-to noise-ratios can be obtained as a no secondary analog circuitry exists in the detector readout path. This system does not have an Airy Scan II 32 channel spatial detector fitted so has no super-resolution capability.
Leica TCS SP8 LSCM
The confocal has two tuneable spectral Hybrid detectors and a spectral tuneable PMT detector. The HyDs can be operated in photon counting or conventional analog mode to record images up to 16 bit depth format. A dedicated transmitted light PMT is also available for acquiring images in bright field format. Pulsed lasers (440 & 470 nm, pulse frequency range 0.3125 to 40 MHz) can be used to perform fluorescence life time imaging including FRET measurement. This system does not have any super-resolution capability.
These instruments provide cutting edge fluorescence microscopy technology to support our novel fluorescence work developed within the School of Molecular Biosciences, (e.g. Spatial Intensity Distribution and Fluorescence Intensity Fluctuation Spectrometry protein homo-oligomerisation analysis, intensity based Fluorescence Resonance Energy Transfer (FRET) imaging, Fluorescence Lifetime FRET imaging, Fluo-8 calcium imaging and FRAP measurement to quantify the diffusion rate and mobile state of fluorescently tagged proteins.
Training
Comprehensive training in the use of the instrumentation will be provided on initial access to equipment and booking system. Staff managing the various rigs will be pleased to give advice on the most appropriate instrumentation to use for a given research project application. The fee for-use basis is to cover the cost of the instrument maintenance contract, staff training time, consumable supplies and the labour costs associated with daily maintenance. If you are applying for research grants we can advise on the charges which are likely to be made over a three-year period. If you require expert assistance and/or training, please kindly plan ahead. We will try out best to answer technical questions on the spot, but users will get out undivided attention if an appointment is made in advance, especially questions regarding experimental designs, potential collaboration, and advanced image analysis.
Booking
The microscopy equipment listed above is available to book, (on a fee for-use basis), by research staff, postgraduate students, final year undergraduate students and commercial clients only after full training has been completed. The same booking criteria is also applicable to staff and students within other Schools.