BioImaging within MCSB
Welcome to our fluorescence imaging web page. 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; Zeiss LSCM 880 and Leica TCS SP8, both invert configuration,which support different areas of fluorescence microscopy.
The Zeiss LSCM 880 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 for transmitted light bright field/DIC imaging. 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 analog circuitry exists in the detector readout path.
The Leica TCS SP8 confocal has two tuneable spectral Hybrid detectors and a spectral tuneable PMT detector. The HyDs can be operated in single photon counting model or conventional analog mode to record images in 16 bit depth format. A dedicated transmitted light PMT is also available for acquiring images in transmitted light bright field/DIC format. These first generation instruments provide the latest cutting edge microscopy technology to support our novel fluorescence work developed within the institute, (e.g. Spatial Intensity Distribution analysis (spIDA), fluoresence resonance energy transfer (FRET), fluorescence life time FRET imaging microscopy, fluorescence recovery after laser photobleaching (FRAP), FLUO-4 calcium imaging).
A third generation old fully motorised Zeiss 510 Pascal Exciter LSCM which provides high resolution 12 bit depth image sizes of up to 2,048 x 2,048 pixels is also available for acquiring multifluorescence z-stack channels over time. Attached lasers can be used to bleach regions of interests within your sample to perform FRAP experiments to quantify the diffusion rate of fluorescently tagged proteins. FRET experiments using the mTeal-mVenus fluorescent protein FRET pair can also be performed using the 458 and 514 nm laser lines.
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 institutes.
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.