Three-dimensional bright-field microscopy with isotropic resolution based on multi-view acquisition and image fusion reconstruction

• Published in: Scientific reports Vol. 10; no. 1; p. 12771
• Main Authors: Calisesi, Gianmaria, Candeo, Alessia, Farina, Andrea, D’Andrea, Cosimo, Magni, Vittorio, Valentini, Gianluca, Pistocchi, Anna, Costa, Alex, Bassi, Andrea
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 29.07.2020; Nature Publishing Group
• Subjects: 639/624/1107/328/2236; 639/624/1107/328/2237; 639/624/1111/55; Anatomy; Aperture; Biological samples; Embryos; Fluorescence; Humanities and Social Sciences; Image processing; Light; Light diffraction; Microscopy; multidisciplinary; Propagation; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-69730-4

Optical Projection Tomography (OPT) is a powerful three-dimensional imaging technique used for the observation of millimeter-scaled biological samples, compatible with bright-field and fluorescence contrast. OPT is affected by spatially variant artifacts caused by the fact that light diffraction is not taken into account by the straight-light propagation models used for reconstruction. These artifacts hinder high-resolution imaging with OPT. In this work we show that, by using a multiview imaging approach, a 3D reconstruction of the bright-field contrast can be obtained without the diffraction artifacts typical of OPT, drastically reducing the amount of acquired data, compared to previously reported approaches. The method, purely based on bright-field contrast of the unstained sample, provides a comprehensive picture of the sample anatomy, as demonstrated in vivo on Arabidopsis thaliana and zebrafish embryos. Furthermore, this bright-field reconstruction can be implemented on practically any multi-view light-sheet fluorescence microscope without complex hardware modifications or calibrations, complementing the fluorescence information with tissue anatomy.