Nanoscale x-ray holo-tomography of human brain tissue with phase retrieval based on multiphoton energy recordings

Abstract

X-ray cone-beam holo-tomography of unstained tissue from the human central nervous system reveals details down to sub-cellular length scales.1 This visualization of variations in the electron density of the sample is based on phase contrast techniques using intensities formed by self-interference of the beam between object and detector. Phase retrieval inverts diffraction and overcomes the phase problem by constraints such as several measurements at different Fresnel numbers for a single projection. Therefore, the object-to-detector distance (defocus) can be varied. However, for cone beam geometry, changing defocus changes magnification, which can be problematic in view of image processing and resolution. Alternatively, the photon energy can be altered (multi-E). Far from absorption edges, multi-E data yield the wavelength independent electron density. In this contribution we present multi-E holo-tomography at the GINIX setup of the P10 beamline at DESY. The instrument is based on a combined optics of elliptical mirrors and an x-ray waveguide positioned in the focal plane for further coherence, spatial Filtering and high numerical aperture.2 Previous results showed the suitability of this instrument for nanoscale tomography of unstained brain tissue.1 We demonstrate that upon energy variation, the focal spot is stable enough for imaging. To this end, a double crystal monochromator and automated alignment routines are required. Three tomograms of human brain tissue were recorded and jointly analyzed using phase retrieval based on the contrast transfer function formalism generalized to multiple photon energies. Variations of the electron density of the sample are successfully reconstructed.