Depth-of-field extension and 3D reconstruction in digital holographic microscopy

Isabelle Bergoënd; Tristan Colomb; Nicolas Pavillon; Yves Emery; Christian Depeursinge

doi:10.1117/12.827350

17 June 2009 Depth-of-field extension and 3D reconstruction in digital holographic microscopy

Isabelle Bergoënd, Tristan Colomb, Nicolas Pavillon, Yves Emery, Christian Depeursinge

Proceedings Volume 7390, Modeling Aspects in Optical Metrology II; 73901C (2009) https://doi.org/10.1117/12.827350
Event: SPIE Europe Optical Metrology, 2009, Munich, Germany

Abstract

The limited depth-of-field is a main drawback of microscopy that prevents from observing, for example, thick semi-transparent objects with all their features in focus. Several algorithms have been developed during the past years to fuse images having various planes of focus and thus obtain a completely focused image with virtually extended depth-of-field. We present a comparison of several of these methods in the particular field of digital holographic microscopy, taking advantage of some of the main properties of holography. We especially study the extended depth-of-field for phase images reconstructed from the hologram of a biological specimen. A criterion of spatial measurement on the object is considered, completed with a visual criterion. The step of distance taken into account to build the stack of images is less than the instrument depth-of-field. Then, preserving the distance of focus associated with each pixel of the image, a three-dimensional representation is presented after automatic detection of the object. The limits of such a method of extraction of 3D information are discussed.

Citation Download Citation

Isabelle Bergoënd, Tristan Colomb, Nicolas Pavillon, Yves Emery, and Christian Depeursinge "Depth-of-field extension and 3D reconstruction in digital holographic microscopy", Proc. SPIE 7390, Modeling Aspects in Optical Metrology II, 73901C (17 June 2009); https://doi.org/10.1117/12.827350

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