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.
Three-dimensional bright-field microscopy with isotropic resolution based on multi-view acquisition and image fusion reconstruction / G. Calisesi, A. Candeo, A. Farina, C. D'Andrea, V. Magni, G. Valentini, A. Pistocchi, A. Costa, A. Bassi. - In: SCIENTIFIC REPORTS. - ISSN 2045-2322. - 10:1(2020), pp. 12771.1-12771.9.
|Titolo:||Three-dimensional bright-field microscopy with isotropic resolution based on multi-view acquisition and image fusion reconstruction|
COSTA, ALEX (Penultimo) [Methodology]
|Settore Scientifico Disciplinare:||Settore BIO/04 - Fisiologia Vegetale|
|Data di pubblicazione:||2020|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1038/s41598-020-69730-4|
|Appare nelle tipologie:||01 - Articolo su periodico|