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International audienceIncreasing the resolution of optical microscopes is a challenging task for studying the cell machinery at the molecular level. 4Pi or TIRF microscopies permit one to reduce the axial dimension of the detection volume. To reduce its lateral dimension, we have proposed a solution in which the scanning head of a 4Pi microscope or of a confocal microscope is coupled to an interferometer. With this technique two beams coming from the source produce two images that are superimposed coherently. For this reason, one can call this technique 2-Image microscopy. It has been shown that with 2-Image microscopy, the complete use of the spatial frequencies collected by the objective allows to reach a 1.22 lambda/4NA lateral resolution. This improvement is independent of the excitation mode and is effective with incoherent light such as fluorescent or chemiluminescent (i.e. without optical excitation) samples. In this paper, we present an interferometric set-up and a modulation technique that make benefit fully from the advantages of 2-Image microscopy

Increasing the lateral resolution of scanning microscopes by a factor of two using 2-Image microscopy