PUBLICATION

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

Authors
Calisesi, G., Candeo, A., Farina, A., D'Andrea, C., Magni, V., Valentini, G., Pistocchi, A., Costa, A., Bassi, A.
ID
ZDB-PUB-200731-16
Date
2020
Source
Scientific Reports   10: 12771 (Journal)
Registered Authors
Keywords
none
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none
PubMed
32728161 Full text @ Sci. Rep.
Abstract
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.
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