Microscope generating a three-dimensional representation of an object and images generated by such a microscope

In a microscope, a method for generating a three-dimensional frequency representation of an observed object, comprising the steps of:

• a) splitting a laser beam into a reference wave and an illuminating wave,
• b) rendering the illuminating wave plane
• c) directing the plane iluminating wave in a first direction for illuminating the observed object, the illuminating wave being diffracted by the object to provide a diffracted wave,
• d) superimposing the diffracted wave and the reference wave to produce an interference pattern,
• e) recording the interference pattern,
• f) calculating from at least the interference pattern a first frequency representation of the diffracted wave corresponding to phases and amplitudes of plane wave components of the diffracted wave,
• g) changing the direction of the illuminating wave and repeating steps (d) to (f) at least once, to produce at least a second frequency representation of the diffracted wave,
• h) applying a first phase shift to the first frequency representation, and
• i) superimposing the first frequency representation and the second frequency representation, to obtain a three-dimensional frequency representation of the observed object,
• wherein the first phase shift is such that the part of the three-dimensional frequency representation corresponding to the first frequency representation has substantially the same phase reference as the part of said three-dimensional frequency representation corresponding to the second frequency representation.

In a microscope, a method for improving an initial three-dimensional frequency representation of an observed object, comprising the steps of:

• a) splitting a laser beam into a reference wave and an illuminating wave,
• b) rendering the illuminating wave plane and directing it in a first direction for illuminating the observed object, thereby providing a diffracted wave diffracted by the object,
• c) superimposing the diffracted wave and the reference wave, to produce an interference pattern,
• d) recording the interference pattern,
• e) calculating, from at least the interference pattern, a first frequency representation of the diffracted wave corresponding to phases and amplitudes of plane wave components of the diffracted wave,
• f) applying a first phase shift to the first frequency representation, and
• g) superimposing the first frequency representation and the initial three-dimensional frequency representation, to obtain a final frequency representation,
• wherein the first phase shift is such that the part of the three-dimensional frequency representation corresponding to the first frequency representation has substantially the same phase reference as the part of said three-dimensional frequency representation corresponding to the second frequency representation.