Super-Resolution Light Microscopy Facility
Super-resolution microscopy is a form of fluorescence light microscopy, which allows the capture of images with a resolution below that of the Abbe diffraction limit. The Super-Resolution Light Microscopy Facility lies within the Wolfson Light Microscopy Facility (LMF) in Firth Court at the University of Sheffield. The suite includes two new super resolution microscopes, one based on structured illumination (OMX SIM, GE Healthcare), and one on localisation microscopy (N STORM, Nikon).
Structured Illumination Microscopy (SIM) uses patterned illumination to generate an image which combines information from the sample and the pattern, bringing small variations in sample fluorescence within the diffraction limit of the microscope. By using multiple different patterns on the same area of sample and with the precise knowledge of what those patterns are, software can be used to reconstruct a super-resolution image. The system is ideal for cultured cells and brightly stained samples and can be used for live cell imaging under certain conditions.
STORM (Stochastic Optical Reconstruction Microscopy) / PALM (Photo-Activated Localisation Microscopy) imaging relies on the specific characteristics of the fluorophores used. Initially, all fluorophores are (effectively) in an 'off' state. When imaging, a random subset of fluorophores are switched on and imaged before being driven back to an 'off' state. As only subsets of fluorophores are imaged at the same time and these are far apart and easily resolvable from each other, their locations can be approximated and recorded. This is carried out for many (>1000) frames allowing a high-resolution image to be formed. This technique is ideal for fixed cells/ molecular structures.
If you wish to collaborate or would like to use a super resolution light microscope, please contact Christa to discuss your requirements.
The Wolfson Light Microscopy Facility
OMX SIM, GE Healthcare
N STORM, Nikon