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

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.

Plant phenotyping relevance

多視点取得と画像融合による3D明視野再構成法を開発し、Arabidopsisの解剖学的形態を実証しているため、植物形態の取得手法が中心です。

Code and data availability

The paper's authors state that the Python sample code implementing their bright-field multi-view reconstruction (used for the Arabidopsis thaliana and zebrafish phenotyping/imaging analysis) is publicly available on GitHub under the authors' account. The GitHub URL in the text contains formatting artifacts and does not