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A clever way to overcome the limitation caused by diffraction is to use 'pointillism strategy', a concept introduced by Rainer Heintzmann in 2005. The idea is to have only one fluorescent emitter (fluorophore) present at a given time for a given pixel. This localisation leads to much higher resolutions. This idea was further developed into what is now known Single Molecule Localization Microscopy (SMLM).
In SMLM, only a sparse subset of the total fluorophore population is emitting at a given time (blinking). This creates a temporal separation of the fluorescence coming from the crowded particles labeling the structure of interest. Single-molecule fluorescence is acquired over thousands of sequential images which will eventually be combined to deliver a super-resolved image with nanometer resolution.
This eBook explores an overview of Super-resolution Microscopy and the concepts behind it.
Download eBookAndor’s microscopy system portfolio provides the most flexible confocal imaging systems for routine to advanced microscopy research. Our cutting-edge Dragonfly systems provide fast and high-resolution imaging in multiple modalities. The BC43 benchtop confocal microscope brings compact high-end imaging at the push of a button.
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