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Super-resolution fluorescence microscopy

Super-resolution in fluorescence microscopy can be defined as all techniques with a resolution higher than the classical diffraction limit of light.

At the Centre for Cellular Imaging there is the ELYRA PS.1 system, which can perform structured illumination microscopy (SIM) and single-molecule localization microscopy (SMLM).

Structured Illumination Microscopy

Structured Illumination Microscopy (SIM) is a technique, which overcomes the diffraction limit by using a patterned illumination and hence creating a moaré effect in the image. An image with twice the resolution in all three spatial dimensions can then be calculated from the known illumination pattern and the recorded image.

The principle of SIM: The sample is illuminated by a striped pattern. Different illumination rotations creates different Moaré patterns in the image.

Single Molecule Localization Microscopy

Single Molecule Localization Microscopy (SMLM) is the common name of several techniques, which utilizes the blinking of fluorophores to surpass the diffraction limit, such as PALM, dSTORM etc. In all of these techniques, a small subset of fluorophores are imaged in each frame of a long time series. The localization of each detected flourophore is found and the final superresolution image is reconstructed from all localization positions.

The principle of SMLM: Instead of collecting the signal from all the labeled molecules in the sample at the same time, like in the widefield (WF) technique, only a few molecules are imaged in each frame of a time series.