The invention describes a simple way of performing Image Scanning Microscopy (ISM) purely optically. It is not necessary to reconstruct the final image from a multitude of individual acquisitions. Instead, the final image is projected directly onto the camera chip. The method is suitable for confocal microscopy as well as for 2-photon microscopy.

Image Scanning Microscopy was developed during the last years and improves the lateral resolution by a factor of 1.6 in comparison with confocal Laser Scanning Microscopy (cLSM). The principal of the method is similar to how the improved resolution in structured illumination microscopy is obtained.

Previously, the disadvantage of ISM was, that for each scan position of the excitation laser an individual image on the camera chip had to be acquired. Subsequently, the final image was calculated by combining all sub-images. Although with this approach ISM was already capable of reaching a significantly higher resolution than cLSM setups, it was also much slower.

New developments have shown that optical ISM without computer aided reconstruction is possible (Optical Photon Reassignment – OPRA). Unfortunately, OPRA demands quite complex optics and it is difficult to upgrade existing cLSM setups. In addition, excitation and emission light pass through the same optics which makes it difficult to apply the method in 2-photon microscopy. 

 How to solve this problem? Read More at MBM ScienceBridge here