Isotropic-Resolution Tomographic Diffractive Microscopy

International audienceMicroscopy techniques based on recording of the optical field diffracted by the specimen, in amplitude and phase, like Digital Holographic Microscopy (DHM) have been a growing research topic in recent years. Tomographic acquisitions are possible if one is able to record information, while controlling variations of the specimen illumination. Classical approaches consider either illumination variation, simple to implement, but suffering fro the classical "missing cone" problem, or sample rotation, delivering images with quasi-isotropic, but lower resolution. We have developed an original-, combined tomographic diffractive microscope setup, making use of specimen rotation as well as illumination rotation, which is able to deliver images with an almost isotropic resolution better than 200 nm

Mécanismes d'usure de revêtements de nitrure de titane et d'aluminium en microtribologie

Le comportement à l’usure de films minces de Ti1-xAlxN (0 ≤ x ≤ 1) déposés par PVD est caractérisé et analysé en fonction de la micro et nano-structure des couches. La structure de ces films est à l’origine de leurs propriétés fonctionnelles. Les directions de croissance des films nanostructurés ont été mesurées par diffraction des rayons X. Les modes d’endommagement des films ont été obtenus par des essais de microtribologie à température ambiante contre une bille d’alumine pour pallier l’usure du pion, à faible chargement et faible vitesse de glissement, pour éviter la formation de couches d’oxyde en surface. Pour comprendre les mécanismes d’usure engendrés par le frottement de la bille, plusieurs essais ont été mis en œuvre : détermination de la ténacité par des essais de rayure, observation de la trace par microscopie électronique à balayage et mesure du volume d’usure après plusieurs allers-retours par profilométrie interférométrique par holographie. Lorsque les débris d’usure restent piégés dans le sillon, leur quantité, directement liée à la résistance à l’initiation de fissures du revêtement, et leur nature, ductiles ou fragiles, ont une grande influence sur l’endommagement final. Les films à forte teneur en aluminium texturés selon la direction [002] du réseau hexagonal, présentent un comportement fragile. Lorsque la teneur en aluminium augmente, le nombre de cycles au bout duquel le coefficient de frottement subit un accroissement considérable diminue. Le nombre de cycles à partir duquel le coefficient de frottement augmente considérablement diminue lorsque la concentration en aluminium augmente. Les films riches en titane dont la direction principale de croissance est [200] du réseau cubique, présentent un meilleur comportement à l’usure. Pour ces films ductiles, l’existence de domaines orientés suivant la direction [111] semble jouer un rôle important sur la ténacité. En effet, la quantité de débris générés est liée à une plus ou moins grande proportion de domaines cristallisant dans cette direction

Tomographic diffractive microscopy: towards highresolution 3-D real-time data acquisition, image reconstruction and display of unlabeled samples

Tomographic diffractive microscopy allows for imaging unlabeled specimens, with a better resolution than conventional microscopes, giving access to the index of refraction distribution within the specimen, and possibly at high speed. Principles of image formation and reconstruction are presented, and progresses towards realtime, three-dimensional acquisition, image reconstruction and final display, are discussed

Roadmap on label-free super-resolution imaging

Label-free super-resolution (LFSR) imaging relies on light-scattering processes in nanoscale objects without a need for fluorescent (FL) staining required in super-resolved FL microscopy. The objectives of this Roadmap are to present a comprehensive vision of the developments, the state-of-the-art in this field, and to discuss the resolution boundaries and hurdles that need to be overcome to break the classical diffraction limit of the label-free imaging. The scope of this Roadmap spans from the advanced interference detection techniques, where the diffraction-limited lateral resolution is combined with unsurpassed axial and temporal resolution, to techniques with true lateral super-resolution capability that are based on understanding resolution as an information science problem, on using novel structured illumination, near-field scanning, and nonlinear optics approaches, and on designing superlenses based on nanoplasmonics, metamaterials, transformation optics, and microsphere-assisted approaches. To this end, this Roadmap brings under the same umbrella researchers from the physics and biomedical optics communities in which such studies have often been developing separately. The ultimate intent of this paper is to create a vision for the current and future developments of LFSR imaging based on its physical mechanisms and to create a great opening for the series of articles in this field

Roadmap on Label-Free Super-resolution Imaging

Label-free super-resolution (LFSR) imaging relies on light-scattering processes in nanoscale objects without a need for fluorescent (FL) staining required in super-resolved FL microscopy. The objectives of this Roadmap are to present a comprehensive vision of the developments, the state-of-the-art in this field, and to discuss the resolution boundaries and hurdles that need to be overcome to break the classical diffraction limit of the label-free imaging. The scope of this Roadmap spans from the advanced interference detection techniques, where the diffraction-limited lateral resolution is combined with unsurpassed axial and temporal resolution, to techniques with true lateral super-resolution capability that are based on understanding resolution as an information science problem, on using novel structured illumination, near-field scanning, and nonlinear optics approaches, and on designing superlenses based on nanoplasmonics, metamaterials, transformation optics, and microsphere-assisted approaches. To this end, this Roadmap brings under the same umbrella researchers from the physics and biomedical optics communities in which such studies have often been developing separately. The ultimate intent of this paper is to create a vision for the current and future developments of LFSR imaging based on its physical mechanisms and to create a great opening for the series of articles in this field.Peer reviewe

High-resolution three-dimensional tomographic diffractive microscopy of transparent inorganic and biological samples

International audienceWe have developed a tomographic diffractive microscope, equipped with a fluorescence confocal scanner. We measure experimentally the lateral resolution using an edge method and by comparing tomographic images of the same samples with wide-field and laser scanning confocal microscopy images; a scanning electron microscope image serves as a reference. The experimental resolution is shown to be to about 130 nm, or / 3.5 NA . This instrument also permits one to measure 3D, complex index of refraction distributions, a quantity that is not accessible to conventional microscopes, and we show how this feature may be used to observe KCl crystals, absorption of which is very weak

Recent Advances and Current Trends in Transmission Tomographic Diffraction Microscopy

Optical microscopy techniques are among the most used methods in biomedical sample characterization. In their more advanced realization, optical microscopes demonstrate resolution down to the nanometric scale. These methods rely on the use of fluorescent sample labeling in order to break the diffraction limit. However, fluorescent molecules’ phototoxicity or photobleaching is not always compatible with the investigated samples. To overcome this limitation, quantitative phase imaging techniques have been proposed. Among these, holographic imaging has demonstrated its ability to image living microscopic samples without staining. However, for a 3D assessment of samples, tomographic acquisitions are needed. Tomographic Diffraction Microscopy (TDM) combines holographic acquisitions with tomographic reconstructions. Relying on a 3D synthetic aperture process, TDM allows for 3D quantitative measurements of the complex refractive index of the investigated sample. Since its initial proposition by Emil Wolf in 1969, the concept of TDM has found a lot of applications and has become one of the hot topics in biomedical imaging. This review focuses on recent achievements in TDM development. Current trends and perspectives of the technique are also discussed

Simplified tomographic diffractive microscopy for axisymmetric samples

International audienceTomographic diffractive microscopy exhibits intrinsic features making it a method of choice for 3D high-resolution label-free imaging. However, these results are achieved at the cost of a heavy data acquisition/reconstruction process. This drawback can be circumvented for certain classes of samples. For example, axisymmetric samples, like optical or textile fibers, present geometrical properties that can be advantageously used to speed-up the acquisition process. We propose to take benefit of these properties to allow for full reconstruction of axisymmetric samples’ complex refractive index distribution, using four approaches, adapted to 3D samples. We applied the proposed reconstruction scheme, based on a numerical rotation of data, to both simulated and experimental data sets

Holographic microscopy and diffractive microtomography of transparent samples

International audienceWe present an optical tomographic diffractive microscope, a device able to image a complex refractive index distribution in three dimensions. Theoretical foundations are first recalled: diffraction under the first Born approximation explains the link between diffracted beam, object frequencies and physical properties of the object. We then describe our experimental setup, recording 2D interferograms in the image space, and detail the image reconstruction process underlying our tomographic microscope, which involves 2D transforms of the recorded interferograms, a peculiar 3D mapping of the data, and a final 3D Fourier reconstruction. We apply tomographic reconstruction to diatom skeletons, unicellular algae with cell walls made of silica, and compare it to holographic reconstruction. We further apply it to pollen grains and show differences between the real and imaginary parts of the measured complex refractive index. Finally, we also recall alternative tomographic methods