3D differential phase contrast microscopy

We demonstrate 3D phase and absorption recovery from partially coherent intensity images captured with a programmable LED array source. Images are captured through-focus with four different illumination patterns. Using first Born and weak object approximations (WOA), a linear 3D differential phase contrast (DPC) model is derived. The partially coherent transfer functions relate the sample's complex refractive index distribution to intensity measurements at varying defocus. Volumetric reconstruction is achieved by a global FFT-based method, without an intermediate 2D phase retrieval step. Because the illumination is spatially partially coherent, the transverse resolution of the reconstructed field achieves twice the NA of coherent systems and improved axial resolution

Redefining A in RGBA: Towards a Standard for Graphical 3D Printing

Advances in multimaterial 3D printing have the potential to reproduce various visual appearance attributes of an object in addition to its shape. Since many existing 3D file formats encode color and translucency by RGBA textures mapped to 3D shapes, RGBA information is particularly important for practical applications. In contrast to color (encoded by RGB), which is specified by the object's reflectance, selected viewing conditions and a standard observer, translucency (encoded by A) is neither linked to any measurable physical nor perceptual quantity. Thus, reproducing translucency encoded by A is open for interpretation. In this paper, we propose a rigorous definition for A suitable for use in graphical 3D printing, which is independent of the 3D printing hardware and software, and which links both optical material properties and perceptual uniformity for human observers. By deriving our definition from the absorption and scattering coefficients of virtual homogeneous reference materials with an isotropic phase function, we achieve two important properties. First, a simple adjustment of A is possible, which preserves the translucency appearance if an object is re-scaled for printing. Second, determining the value of A for a real (potentially non-homogeneous) material, can be achieved by minimizing a distance function between light transport measurements of this material and simulated measurements of the reference materials. Such measurements can be conducted by commercial spectrophotometers used in graphic arts. Finally, we conduct visual experiments employing the method of constant stimuli, and derive from them an embedding of A into a nearly perceptually uniform scale of translucency for the reference materials.Comment: 20 pages (incl. appendices), 20 figures. Version with higher quality images: https://cloud-ext.igd.fraunhofer.de/s/pAMH67XjstaNcrF (main article) and https://cloud-ext.igd.fraunhofer.de/s/4rR5bH3FMfNsS5q (appendix). Supplemental material including code: https://cloud-ext.igd.fraunhofer.de/s/9BrZaj5Uh5d0cOU/downloa

Confocal Microscopy and Three-Dimensional Reconstruction of Thick, Transparent, Vital Tissue

The three-dimensional visualization of the 400 micron thick, transparent, in situ cornea is described to demonstrate the use of confocal light microscopy for noninvasive imaging of living cells and thick tissues in their normal, vital conditions. Specimen preparation and physiological stability, as well as light attenuation corrections are critical to data acquisition. The technique to provide mechanical stability of the specimen during the duration of the image acquisition is explained. A laser scanning confocal light microscope (LSCM) was used to obtain optical serial sections from rabbit eyes that were freshly removed and placed in a physiological Ringer\u27s solution. This study demonstrates the capability of the confocal light microscope to obtain a series of high contrast images, with a depth resolution of one micron, across the full thickness of living, transparent tissue. The problems of nonisotropic sampling and the limited eight-bit dynamic range are discussed. The three-dimensional reconstructions were obtained by computer graphics using the volume visualization projection technique. The three-dimensional visualization of the cornea in the in situ eye is presented as an example of image understanding of thick, viable biological cells and tissues. Finally, the criterion of image fidelity is explained. The techniques of confocal light microscopy with its enhanced lateral and axial resolution, improved image contrast, and volume visualization provides microscopists with new techniques for the observation of vital cells and tissues, both in vivo and in vitro