Chapter 4
Tomographic Diffractive Microscopy:
Principles, Implementations,
and Applications in Biology
Bertrand Simon and Olivier Haeberlé
Abstract Tomographic Diffractive Microscopy (TDM) is an advanced digital
microscopic imaging technique, extending the capabilities of Digital Holographic
Microscopy (DHM), which delivers 3D quantitative images of the index of refraction distribution within the observed sample. It is a two-step imaging approach based
first on recording of multiple holograms under varying conditions of illumination, and
second on applying sample-adapted numerical inversion procedures to reconstruct
the 3-D image of the specimen under consideration. After a short recall of DHM
principles, applications, and limitations, the principle of TDM is introduced, and
then several of its practical implementations are described. We put emphasis on their
respective advantages and limitations, through various examples of application for
biological samples imaging. Perspectives, and challenges for this imaging modality,
as well as some hints to address them, are finally presented in the conclusion.
4.1 Introduction
Its unique capabilities for imaging living specimens, in three dimensions, possibly
over long periods make the optical microscope an invaluable tool for biological
research. This has motivated, since its invention and its adoption as a scientific tool,
the development of numerous techniques to improve the contrast and visibility of
the observed sample, through, for example, oblique and Rheinberg illuminations,
dark-field microscopy, phase contrast and differential interference contrast (DIC),
Dodt gradient contrast, Hoffman modulation, or polarized microscopy [1]. All these
B. Simon (B) · O. Haeberlé
Laboratoire Photonique Numérique et Nanosciences (LP2N - UMR 5298), Institut d’Optique
Graduate School, Université de Bordeaux, CNRS, Bordeaux, France
e-mail: bertrand.simon@institutoptique.fr
O. Haeberlé
e-mail: olivier.haeberle@uha.fr
Institut de Recherche en Informatique, Mathématiques, Automatique et Signal (IRIMAS -
EA7499), Université de Haute-Alsace, Mulhouse, France
© Springer Nature Switzerland AG 2019
V. Astratov (ed.), Label-Free Super-Resolution Microscopy,
Biological and Medical Physics, Biomedical Engineering,
https://doi.org/10.1007/978-3-030-21722-8_4
85
Tomographic Diffractive Microscopy:
Principles, Implementations,
and Applications in Biology
Bertrand Simon and Olivier Haeberlé
Abstract Tomographic Diffractive Microscopy (TDM) is an advanced digital
microscopic imaging technique, extending the capabilities of Digital Holographic
Microscopy (DHM), which delivers 3D quantitative images of the index of refraction distribution within the observed sample. It is a two-step imaging approach based
first on recording of multiple holograms under varying conditions of illumination, and
second on applying sample-adapted numerical inversion procedures to reconstruct
the 3-D image of the specimen under consideration. After a short recall of DHM
principles, applications, and limitations, the principle of TDM is introduced, and
then several of its practical implementations are described. We put emphasis on their
respective advantages and limitations, through various examples of application for
biological samples imaging. Perspectives, and challenges for this imaging modality,
as well as some hints to address them, are finally presented in the conclusion.
4.1 Introduction
Its unique capabilities for imaging living specimens, in three dimensions, possibly
over long periods make the optical microscope an invaluable tool for biological
research. This has motivated, since its invention and its adoption as a scientific tool,
the development of numerous techniques to improve the contrast and visibility of
the observed sample, through, for example, oblique and Rheinberg illuminations,
dark-field microscopy, phase contrast and differential interference contrast (DIC),
Dodt gradient contrast, Hoffman modulation, or polarized microscopy [1]. All these
B. Simon (B) · O. Haeberlé
Laboratoire Photonique Numérique et Nanosciences (LP2N - UMR 5298), Institut d’Optique
Graduate School, Université de Bordeaux, CNRS, Bordeaux, France
e-mail: bertrand.simon@institutoptique.fr
O. Haeberlé
e-mail: olivier.haeberle@uha.fr
Institut de Recherche en Informatique, Mathématiques, Automatique et Signal (IRIMAS -
EA7499), Université de Haute-Alsace, Mulhouse, France
© Springer Nature Switzerland AG 2019
V. Astratov (ed.), Label-Free Super-Resolution Microscopy,
Biological and Medical Physics, Biomedical Engineering,
https://doi.org/10.1007/978-3-030-21722-8_4
85