|Confocal:||Confocal imaging is a fluorescence microscopy technique that optically sections the speciment preventing out of focus light from reaching the detector. This yields clear high contrast images, and together with the ability to acquire images sequentially at multiple positions the entire sample can be reconstructed in 3D.|
|Live Cell Imaging:||Many biological processes are dynamic events and as such observing living cells is crucial to unravelling and understanding the mechanisms behind them. This requires special consideration when imaging. Firstly, the environmental conditions must be controllable and stable to ensure the cell remain healthy. Secondly, care needs to be taken that the act of imaging has as little impact as possible on the event being observed.|
|STED:||Stimulated Emission Depletion (STED) is a super resolution fluorescent microscope technique designed to enhance image resolution by bypassing the diffraction limit faced by tranditional microscopy.|
FRET (Fluorescence Resonance Energy Transfer) is a technique used largely to investigate molecular interactions, or with specially designed reports it can show if molecules are in an active or inactive state. It works by using two carefully selected fluorophores that when in close enough proximity and suitable orientation energy from the donor fluorophore can be transferred to the accepter. This transfer can be observed by monitoring the acceptors emission or the donors fluorescence lifetime.
Fluorescence Recovery After Photobleaching is used to gain insight into the dynamics or both diffusion and binding rates within a sample. Photobleaching is an irreversible process by which a fluorophore loses the ability to absorb and emit light. By deliberately inducing photobleaching within a restricted region of the sample observing the recovery within this region over time is affected by the rate at which unbleached molecules entre this region and bleached molecules are moved. The light sheet mode available on this instrument allows to study thicker samples than (<2mm) what an ordinary confocal microscope could image
Fluorescence Correlating Spectroscopy allows for determining diffusion coefficients and concentrations of fluorescently labelled molecules at nanomolar concentrations both in vitro and in live cells. It is mostly useful for indirect studies of molecular activity in plasma membrane, in cytosol and in nucleus via following relative changes in diffusive behavior and/or concentration.
Light microscopy lends itself very well to labelling multiple structures within a sample due to the ability to separate these spatially overlapping signals by the wavelength of light they emit. Colocalisation is the study of how the distribution of one probe relates to that of another within the same sample.